U.S. patent application number 12/939778 was filed with the patent office on 2011-11-10 for hepatitis c virus inhibitors.
This patent application is currently assigned to Bristol-Myers Squibb Company. Invention is credited to Makonen Belema, John A. Bender, Qi Chen, Piyasena Hewawasam, Rico Lavoie, Omar D. Lopez, Gan Wang, Zhong Yang.
Application Number | 20110274648 12/939778 |
Document ID | / |
Family ID | 43466793 |
Filed Date | 2011-11-10 |
United States Patent
Application |
20110274648 |
Kind Code |
A1 |
Lavoie; Rico ; et
al. |
November 10, 2011 |
Hepatitis C Virus Inhibitors
Abstract
The present disclosure relates to compounds, compositions and
methods for the treatment of Hepatitis C virus (HCV) infection.
Also disclosed are pharmaceutical compositions containing such
compounds and methods for using these compounds in the treatment of
HCV infection.
Inventors: |
Lavoie; Rico; (Candiac,
CA) ; Bender; John A.; (Middletown, CT) ;
Yang; Zhong; (Southington, CT) ; Belema; Makonen;
(North Haven, CT) ; Lopez; Omar D.; (Wallingford,
CT) ; Chen; Qi; (Stamford, CT) ; Wang;
Gan; (Wallingford, CT) ; Hewawasam; Piyasena;
(Middletown, CT) |
Assignee: |
Bristol-Myers Squibb
Company
|
Family ID: |
43466793 |
Appl. No.: |
12/939778 |
Filed: |
November 4, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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61378806 |
Aug 31, 2010 |
|
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61260115 |
Nov 11, 2009 |
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Current U.S.
Class: |
424/85.2 ;
424/85.4; 424/85.7; 514/232.2; 514/249; 514/300; 514/303; 514/306;
514/316; 514/394; 514/397; 514/43; 514/44A; 544/350; 544/353;
544/79; 546/118; 546/122; 546/173; 546/187; 548/305.4; 548/306.1;
548/313.1 |
Current CPC
Class: |
A61K 31/5377 20130101;
A61K 31/4184 20130101; A61K 31/4375 20130101; C07D 403/14 20130101;
A61K 31/4178 20130101; C07D 309/06 20130101; C07D 405/14 20130101;
A61P 43/00 20180101; A61P 1/16 20180101; C07D 413/14 20130101; C07D
403/10 20130101; A61K 31/437 20130101; C07D 471/04 20130101; C07D
401/14 20130101; A61P 31/14 20180101; A61K 45/06 20130101; A61K
31/4709 20130101; C07D 487/04 20130101; A61K 31/4545 20130101; A61K
31/498 20130101; A61K 31/4985 20130101 |
Class at
Publication: |
424/85.2 ;
548/305.4; 514/394; 548/306.1; 546/187; 514/316; 546/118; 514/303;
544/350; 514/249; 548/313.1; 514/397; 546/173; 514/306; 544/353;
546/122; 514/300; 544/79; 514/232.2; 424/85.4; 514/43; 424/85.7;
514/44.A |
International
Class: |
A61K 31/4184 20060101
A61K031/4184; C07D 401/14 20060101 C07D401/14; A61K 31/4545
20060101 A61K031/4545; A61K 31/437 20060101 A61K031/437; A61K
31/4985 20060101 A61K031/4985; A61K 31/4178 20060101 A61K031/4178;
A61K 31/4709 20060101 A61K031/4709; C07D 471/04 20060101
C07D471/04; A61K 31/4375 20060101 A61K031/4375; C07D 413/14
20060101 C07D413/14; A61K 31/5377 20060101 A61K031/5377; A61K 38/21
20060101 A61K038/21; A61K 31/7056 20060101 A61K031/7056; A61K 38/20
20060101 A61K038/20; A61K 31/713 20060101 A61K031/713; A61P 31/14
20060101 A61P031/14; C07D 403/14 20060101 C07D403/14 |
Claims
1. A compound of Formula (I) ##STR00752## or a pharmaceutically
acceptable salt thereof, wherein: X and X' are each independently
selected from CH, CR.sup.1, and N; Y and Y' are each independently
selected from CH, CR.sup.2, and N; provided that no more than two
of X, X', Y, and Y' are N; R.sup.1 and R.sup.2 are independently
selected from ##STR00753## ##STR00754## each R.sup.3 is
independently selected from hydrogen, cyano, and halo; each R.sup.4
is independently selected from hydrogen, and alkyl, wherein the
alkyl can optionally form a fused three- to five-membered ring with
an adjacent carbon atom wherein said ring is optionally substituted
with one or two methyl groups; or, R.sup.4 and the carbon to which
it is attached form an ethylene group; each R.sup.5 is
independently selected from hydrogen and --C(O)R.sup.6; each
R.sup.6 is independently selected from alkoxy, alkyl, arylalkoxy,
arylalkyl, cycloalkyl, heterocyclyl, heterocyclylalkyl,
(NR.sup.cR.sup.d)alkenyl, and (NR.sup.cR.sup.d)alkyl; and each
R.sup.7 and R.sup.8 is independently selected from hydrogen and
alkyl.
2. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein: X and X' are independently selected from CH and
CR.sup.1; and Y, and Y' are independently selected from CH and
CR.sup.2.
3. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein: X and X' are independently selected from CH and
CR.sup.1; and Y and Y' are each N.
4. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein: Y and X' are each N; X is selected from CH and
CR.sup.1; and Y' is selected from CH and CR.sup.2.
5. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein: X' is N; and X is selected from CH and CR.sup.1;
and Y and Y' are independently selected from CH and CR.sup.2.
6. A compound selected from: methyl
((1S)-1-(((2S)-2-(5-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimida-
zol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimida-
zol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-(1-((2S)-2-((methoxycarbonyl)amino)-3-methyl-
butanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)phenyl)-1H-imid-
azol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;
methyl
((1S)-1-(((2S)-2-(4-(4-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)phenyl)-1H-
-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;
methyl
((1S)-2-((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrroli-
dinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidi-
nyl)-1-methyl-2-oxoethyl)carbamate; dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrrolidinediyl-
((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate;
5,5'-(2,6-naphthalenediyl)bis(2-(2S)-1-((2R)-2-phenyl-2-(1-piperidinyl)ac-
etyl)-2-pyrrolidinyl)-1H-benzimidazole);
(1R)-2-((2S)-2-(5-(6-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-
-pyrrolidinyl)-1H-benzimidazol-6-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-p-
yrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine; methyl
((1S,2R)-2-methoxy-1-(((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-.beta-
.-methyl-L-threonyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H--
benzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate; methyl
((1S)-3-methoxy-1-(((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-.beta.-m-
ethyl-L-homoseryl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-be-
nzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate; dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrrolidinediyl-
((1S)-1-cyclopropyl-2-oxo-2,1-ethanediyl)))biscarbamate; dimethyl
(2,7-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrrolidinediyl-
((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate; methyl
((1S)-2-((2S)-2-(5-(7-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrroli-
dinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidi-
nyl)-1-methyl-2-oxoethyl)carbamate; methyl
((1R)-1-(((2S)-2-(5-(7-(2-((2S)-1-((2R)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimida-
zol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;
5,5'-(2,7-naphthalenediyl)bis(2-(2S)-1-((2R)-2-phenyl-2-(1-piperidinyl)ac-
etyl)-2-pyrrolidinyl)-1H-benzimidazole); methyl
((1S)-1-(((2S)-2-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol--
2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((2S)-2-(4-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol--
2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate;
(1R)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2R)-2-(diethylamino)-2-phen-
ylacetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-2-naphthyl)-1-
H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-N,N-diethyl-2-oxo-1-phen-
ylethanamine; methyl
((1R)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-phenylacetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2--
naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-pheny-
lethyl)carbamate; dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabicyclo[-
3.1.0]hexane-3,2-diyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate;
methyl
((1S)-1-(((2S)-2-(6-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-
-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyridin-6-yl)-2-naphth-
yl)-1H-imidazo[4,5-b]pyridin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl-
)carbamate; methyl
((1S)-1-(((2R)-2-(6-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyridin-6-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyridin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate; methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate; methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2R)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate; methyl
((1S)-1-(((2R)-2-(5-(6-(2-((2R)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate; dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrrolidinediyl-
((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanediyl)))biscarbamate;
methyl
((1S)-2-((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-ben-
zimidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)ca-
rbamate; methyl
((1S)-2-((2S)-2-(4-(6-(2-(2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-(tetra-
hydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphth-
yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)et-
hyl)carbamate; methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imi-
dazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;
dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabicyclo[-
3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanedi-
yl)))biscarbamate; methyl
((1S)-1-(((2S)-2-(4-((6-((2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-me-
thylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)ethynyl-
)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;
methyl
((1S)-1-(((2S)-2-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-
-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)-1H-imidaz-
ol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-(t-
etrahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphth-
yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran--
4-yl)ethyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)--
2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-2-quinolinyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-2-quinoxalinyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbony-
l)-2-methylpropyl)carbamate; dimethyl
(2,6-quinoxalinediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabicyclo[-
3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanedi-
yl)))biscarbamate; dimethyl
(2,6-quinolinediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabicyclo[3.-
1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanediyl-
)))biscarbamate; methyl
((1S)-2-((2S)-2-(4-chloro-5-(4-(6-(2-(2S)-1-((2S)-2-((methoxycarbonyl)ami-
no)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)--
2-naphthyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2-
H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((2S)-2-(4-(6-(4-(4-chloro-2-((2S)-1-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)phenyl)-2-naphthy-
l)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;
methyl
((1S)-2-((1R,3S,5R)-3-(7-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-y-
l)-1H-imidazol-4-yl)-2-naphthyl)-1H-naphtho[1,2-d]imidazol-2-yl)-2-azabicy-
clo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate;
methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-
-3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-quinolinyl)-1H--
benzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate;
methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4--
yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl-
)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-b-
enzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-y-
l)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6'-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)-
amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl-
)-2,2'-binaphthalen-6-yl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-y-
l)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-1,5-naphthyridin-2-yl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-
carbonyl)-2-methylpropyl)carbamate; dimethyl
(1,5-naphthyridine-2,6-diylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azab-
icyclo[3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-e-
thanediyl)))biscarbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbon-
yl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-
-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate;
methyl
((1S)-1-(((1S,3R,5S)-3-(4-(6-(3-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)p-
henyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(2-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-6-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(2-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-b-
enzimidazol-5-yl)-6-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-na-
phthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrah-
ydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2--
yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-
-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo--
1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex--
3-yl)-1H-imidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-b-
enzimidazol-5-yl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)--
2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)--
1H-imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]-
hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ph-
enyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(-
tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-
carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-
carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-
-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(4,4-difluorocyclohexyl)-2-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)--
2-((2S)-2-(4,4-difluorocyclohexyl)-2-((methoxycarbonyl)amino)acetyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-
-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxoethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)butanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthy-
l)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)propyl)ca-
rbamate; methyl
((1S)-1-(((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]h-
ex-3-yl)-1H-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]h-
ex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-
-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-2-oxo-
-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(4,4-difluorocyclohexyl)-2-((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S-
)-2-(4,4-difluorocyclohexyl)-2-((methoxycarbonyl)amino)acetyl)-5-methyl-2--
pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-met-
hyl-1-pyrrolidinyl)-2-oxoethyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(4-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-
-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2--
azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbama-
te; methyl
((1S,2R)-2-methoxy-14(1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-(N--
(methoxycarbonyl)-O-methyl-L-threonyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imi-
dazol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2--
yl)carbonyl)propyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethy-
nyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate; methyl
(2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-(((methoxycarbonyl)amino)(tetrah-
ydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)e-
thynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-
-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-((4-chloro-2-((1R,3S,5R)-2-((2S)-2--
((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-
-imidazol-5-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(((2R,4r,6S)-2,6-dimethy-
ltetrahydro-2H-pyran-4-yl)((methoxycarbonyl)amino)acetyl)-2-azabicyclo[3.1-
.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azab-
icyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6'-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)-
amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2,-
2'-binaphthalen-6-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbon-
yl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,5R)-2-((2S)-2-((2R,4r,6S)-2,6-dim-
ethyltetrahydro-2H-pyran-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-azabicy-
clo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-
-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate;
methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)p-
henyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbony-
l)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0-
]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate;
methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
x-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-2-((1R,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tet-
rahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)--
2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)--
1H-imidazol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-y-
l)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-
-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)--
2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carba-
mate; methyl
((1S)-1-(((2S,5S)-2-(4-chloro-5-(4-(6-(4-chloro-2-((2S,5S)-1-((2S)-2-((me-
thoxycarbonyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazo-
l-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbo-
nyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(5-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)--
1-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(4-(5-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)-1-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((1R,3S,5R)-3-
-(4-((6-(2-((1R,3S,5R)-2-((2S)-2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyr-
an-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-
-imidazol-4-yl)-2-naphthyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxoethyl)carbamate; methyl
((1S)-1-(((2S,3aS,6aS)-2-(4-(6-(4-(2-((2S,3aS,6aS)-1-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)octahydrocyclopenta[b]pyrrol-2-yl)-1H-imidaz-
ol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)hexahydrocyclopenta[b]pyrrol--
1(2H)-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((2S,3aS,6aS)-2-(4-(4-(6-(2-((2S,3aS,6aS)-1-((2S)-2-((methoxycarb-
onyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)hexahydrocyclopenta[b]pyrro-
l-2(2H)-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)hexahydr-
ocyclopenta[b]pyrrol-1(2H)-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)car-
bamate;
3-chloro-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((3-chloro-5-met-
hoxy-1-isoquinolinyl)carbonyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4--
yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carb-
onyl)-5-methoxyisoquinoline; methyl
((1S)-1-(((2S,5S)-2-(4-(4-(6-(4-chloro-2-((2S,5S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-5-yl)-2-
-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate; methyl
((1S)-1-(((2S,5S)-2-(4-chloro-5-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-
-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-((6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-n-
aphthyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(t-
etrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((2R,4r,6S)-2,6-di-
methyltetrahydro-2H-pyran-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-azabic-
yclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-y-
l)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate;
methyl
((1S)-2-((2S,5S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imi-
dazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-
-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-((6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthy-
l)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methy-
lpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethy-
nyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(t-
etrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethy-
nyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-((-
3S)-tetrahydro-2H-pyran-3-yl)ethyl)carbamate; methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-2-q-
uinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methylpro-
pyl)carbamate; methyl
((1S)-1-(((2S,5S)-2-(4-(6-(4-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-2-q-
uinolinyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methylpro-
pyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)p-
henyl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbo-
nyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)phenyl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate;
methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-((4-chloro-2-((1R,3S,5R)-2-((2S)-2-(-
(methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3-
.1.0]hex-3-yl)-1H-imidazol-5-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-a-
zabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamat-
e; methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)-
amino)-3-methylbutanoyl)amino)-2-methylpropyl)-1H-imidazol-4-yl)-2-naphthy-
l)phenyl)-1H-imidazol-2-yl)-2-methylpropyl)carbamoyl)-2-methylpropyl)carba-
mate;
3-methyl-N-((1S)-2-methyl-1-(4-(4-(6-(2-((1S)-2-methyl-1-((3-methylb-
utanoyl)amino)propyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-y-
l)propyl)butanamide;
2-((2S,5S)-1-acetyl-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-1-acetyl-
-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
e; methyl
(1-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((2S,5S)--
2-(4-(4-(6-(2-((2S,5S)-1-(((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl-
)((methoxycarbonyl)amino)acetyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl-
)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-2-oxoethyl-
)carbamate; methyl
((1S)-2-((2S,4S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imi-
dazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-
-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((2S,4S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethyny-
l)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methy-
lpropyl)carbamate;
2-((2S,4S)-4-methyl-1-(3-methylbutanoyl)-2-pyrrolidinyl)-4-(4-(6-(2-((2S,-
4S)-4-methyl-1-(3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naph-
thyl)phenyl)-1H-imidazole; methyl
((1S)-2-((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1((2S)-2-((methoxycarbonyl)amino)-
-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol--
4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-ox-
o-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((3R)-3-(4-(4-(6-(2-((3R)-4-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-3-morpholinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imid-
azol-2-yl)-4-morpholinyl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((3R)-3-(4-(4-(6-(2-((3R)-4-((2S)-2-((methoxycarbonyl)amino)-2-(t-
etrahydro-2H-pyran-4-yl)acetyl)-3-morpholinyl)-1H-imidazol-4-yl)-2-naphthy-
l)phenyl)-1H-imidazol-2-yl)-4-morpholinyl)-2-oxo-1-(tetrahydro-2H-pyran-4--
yl)ethyl)carbamate;
(1R,3S,5R)-2-(3-methylbutanoyl)-3-(4-(6-((2-((1R,3S,5R)-2-(3-methylbutano-
yl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H--
imidazol-2-yl)-2-azabicyclo[3.1.0]hexane; methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-2-piperidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imid-
azol-2-yl)-1-piperidinyl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-(t-
etrahydro-2H-pyran-4-yl)acetyl)-2-piperidinyl)-1H-imidazol-4-yl)-2-naphthy-
l)phenyl)-1H-imidazol-2-yl)-1-piperidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4--
yl)ethyl)carbamate; methyl
((1S)-1-(((2S,5R)-2-(4-(4-(6-(2-((2S,5R)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate; methyl
((1S)-2-((2S,5R)-2-(4-(4-(6-(2-((2S,5R)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-
-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-2-oxo-
-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((2S)-2-(4-(6-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-4-methylene-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-2-naph-
thyl)-1H-imidazol-2-yl)-4-methylene-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate; methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-(t-
etrahydro-2H-pyran-4-yl)acetyl)-4-methylene-2-pyrrolidinyl)-1H-imidazol-4--
yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methylene-1-pyrrolidinyl)-2-oxo-
-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-3--
methylbutanoyl)amino)ethyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidaz-
ol-2-yl)ethyl)carbamoyl)-2-methylpropyl)carbamate; methyl
((1S)-2-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-2--
(tetrahydro-2H-pyran-4-yl)acetyl)amino)ethyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)ethyl)amino)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)e-
thyl)carbamate; methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-3--
methylbutanoyl)(methyl)amino)ethyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1-
H-imidazol-2-yl)ethyl)(methyl)carbamoyl)-2-methylpropyl)carbamate;
methyl
((1S)-2-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-2--
(tetrahydro-2H-pyran-4-yl)acetyl)(methyl)amino)ethyl)-1H-imidazol-4-yl)-2--
naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)(methyl)amino)-2-oxo-1-(tetrahydro-
-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate; methyl
(2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(((methoxycarbonyl)amino)(tetrahydro-
-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphth-
yl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxo-1-(tetrahydro--
2H-pyran-4-yl)ethyl)carbamate; methyl
(1-cyclopropyl-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(cyclopropyl((methoxyc-
arbonyl)amino)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthy-
l)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxoethyl)carbamate;
methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbon-
yl)amino)butanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)p-
henyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)propyl)carbamate-
; methyl
((1S)-1-cyclobutyl-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-cy-
clobutyl-2-((methoxycarbonyl)amino)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imi-
dazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)--
2-oxoethyl)carbamate; methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-((2R,4S)-2-methyltetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidi-
nyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyr-
rolidinyl)-1-((2S,4R)-2-methyltetrahydro-2H-pyran-4-yl)-2-oxoethyl)carbama-
te; methyl
((1S)-1-(((2S,4R)-2-(4-(4-(6-(2-((2S,4R)-1-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)--
2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-m-
ethylpropyl)carbamate; methyl
((1S)-1-(((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)--
2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate; methyl
((1S)-1-(((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo[3.1.0]-
hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-azabicyclo[3.1.0]he-
x-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2--
azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbama-
te; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((meth-
oxycarbonyl)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-
-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicy-
clo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo[3.1.0]-
hex-2-yl)carbonyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazo-
l-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbo-
nyl)-2-methylpropyl)carbamate; methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amino-
)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)-
phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H--
pyran-4-yl)ethyl)carbamate; and methyl
((1S)-1-(((2S,4S)-2-(4-chloro-5-(6-(4-(4-chloro-2-((2S,4S)-1-((2S)-2-((me-
thoxycarbonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazo-
l-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbo-
nyl)-2-methylpropyl)carbamate; methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(4-chloro-2-((2S,4S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-5-yl)-2-
-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate; methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(4-chloro-2-((2S,4S)-1-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-
-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidin-
yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(4-chloro-2-((2S,4S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-5-yl)ph-
enyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate; methyl
(2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(((methoxycarbonyl)amino)(tetrahydro-
-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphth-
yl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxo-1-(tetrahydro--
2H-pyran-4-yl)ethyl)carbamate; methyl
((1S)-2-((5S)-5-(4-(4-(6-(2-((3S)-2-((2S)-2-((methoxycarbonyl)amino)-2-(t-
etrahydro-2H-pyran-4-yl)acetyl)-1-methyl-3-pyrazolidinyl)-1H-imidazol-4-yl-
)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-methyl-1-pyrazolidinyl)-2-oxo-1-(-
tetrahydro-2H-pyran-4-yl)ethyl)carbamate; methyl
((1R)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2R)-2-((methoxycarbonyl)amino)-3--
methylbutanoyl)amino)-2,2-dimethylpropyl)-1H-imidazol-4-yl)-2-naphthyl)phe-
nyl)-1H-imidazol-2-yl)-2,2-dimethylpropyl)carbamoyl)-2-methylpropyl)carbam-
ate; methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-3--
methylbutanoyl)amino)-2,2-dimethylpropyl)-1H-imidazol-4-yl)-2-naphthyl)phe-
nyl)-1H-imidazol-2-yl)-2,2-dimethylpropyl)carbamoyl)-2-methylpropyl)carbam-
ate; and
N-((1S)-1-(4-(4-(6-(2-((1S)-2,2-dimethyl-1-((3-methylbutanoyl)ami-
no)propyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2,2-dime-
thylpropyl)-3-methylbutanamide); or a pharmaceutically acceptable
salt thereof.
7. A composition comprising a compound of claim 1, or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
8. The composition of claim 7 further comprising at least one
additional compound having anti-HCV activity.
9. The composition of claim 8 wherein at least one of the
additional compounds is an interferon or a ribavirin.
10. The composition of claim 9 wherein the interferon is selected
from interferon alpha 2B, pegylated interferon alpha, consensus
interferon, interferon alpha 2A, and lymphoblastoid interferon
tau.
11. The composition of claim 8 wherein at least one of the
additional compounds is selected from interleukin 2, interleukin 6,
interleukin 12, a compound that enhances the development of a type
1 helper T cell response, interfering RNA, anti-sense RNA,
Imiqimod, ribavirin, an inosine 5'-monophospate dehydrogenase
inhibitor, amantadine, and rimantadine.
12. The composition of claim 8 wherein at least one of the
additional compounds is effective to inhibit the function of a
target selected from HCV metalloprotease, HCV serine protease, HCV
polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV
assembly, HCV egress, HCV NS5A protein, and IMPDH for the treatment
of an HCV infection.
13. A method of treating an HCV infection in a patient, comprising
administering to the patient a therapeutically effective amount of
a compound of claim 1, or a pharmaceutically acceptable salt
thereof.
14. The method of claim 13 further comprising administering at
least one additional compound having anti-HCV activity prior to,
after or simultaneously with the compound of claim 1, or a
pharmaceutically acceptable salt thereof.
15. The method of claim 14 wherein at least one of the additional
compounds is an interferon or a ribavirin.
16. The method of claim 15 wherein the interferon is selected from
interferon alpha 2B, pegylated interferon alpha, consensus
interferon, interferon alpha 2A, and lymphoblastoid interferon
tau.
17. The method of claim 14 wherein at least one of the additional
compounds is selected from interleukin 2, interleukin 6,
interleukin 12, a compound that enhances the development of a type
1 helper T cell response, interfering RNA, anti-sense RNA,
Imiqimod, ribavirin, an inosine 5'-monophospate dehydrogenase
inhibitor, amantadine, and rimantadine.
18. The method of claim 14 wherein at least one of the additional
compounds is effective to inhibit the function of a target selected
from HCV metalloprotease, HCV serine protease, HCV polymerase, HCV
helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress,
HCV NS5A protein, and IMPDH for the treatment of an HCV infection.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 61/260,115 filed Nov. 11, 2009 and U.S. Provisional
Application No. 61/378,806 filed Aug. 31, 2010, incorporated herein
by reference in their entirety.
[0002] The present disclosure is generally directed to antiviral
compounds, and more specifically directed to compounds which can
inhibit the function of the NS5A protein encoded by Hepatitis C
virus (HCV), compositions comprising such compounds, and methods
for inhibiting the function of the NS5A protein.
[0003] HCV is a major human pathogen, infecting an estimated 170
million persons worldwide--roughly five times the number infected
by human immunodeficiency virus type 1. A substantial fraction of
these HCV infected individuals develop serious progressive liver
disease, including cirrhosis and hepatocellular carcinoma.
[0004] The current standard of care for HCV, which employs a
combination of pegylated-interferon and ribavirin, has a
non-optimal success rate in achieving sustained viral response and
causes numerous side effects. Thus, there is a clear and long-felt
need to develop effective therapies to address this undermet
medical need.
[0005] HCV is a positive-stranded RNA virus. Based on a comparison
of the deduced amino acid sequence and the extensive similarity in
the 5' untranslated region, HCV has been classified as a separate
genus in the Flaviviridae family. All members of the Flaviviridae
family have enveloped virions that contain a positive stranded RNA
genome encoding all known virus-specific proteins via translation
of a single, uninterrupted, open reading frame.
[0006] Considerable heterogeneity is found within the nucleotide
and encoded amino acid sequence throughout the HCV genome due to
the high error rate of the encoded RNA dependent RNA polymerase
which lacks a proof-reading capability. At least six major
genotypes have been characterized, and more than 50 subtypes have
been described with distribution worldwide. The clinical
significance of the genetic heterogeneity of HCV has demonstrated a
propensity for mutations to arise during monotherapy treatment,
thus additional treatment options for use are desired. The possible
modulator effect of genotypes on pathogenesis and therapy remains
elusive.
[0007] The single strand HCV RNA genome is approximately 9500
nucleotides in length and has a single open reading frame (ORF)
encoding a single large polyprotein of about 3000 amino acids. In
infected cells, this polyprotein is cleaved at multiple sites by
cellular and viral proteases to produce the structural and
non-structural (NS) proteins. In the case of HCV, the generation of
mature non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, and
NS5B) is effected by two viral proteases. The first one is believed
to be a metalloprotease and cleaves at the NS2-NS3 junction; the
second one is a serine protease contained within the N-terminal
region of NS3 (also referred to herein as NS3 protease) and
mediates all the subsequent cleavages downstream of NS3, both in
cis, at the NS3-NS4A cleavage site, and in trans, for the remaining
NS4A-NS4B, NS4B-NS5A, NS5A-NS5B sites. The NS4A protein appears to
serve multiple functions by both acting as a cofactor for the NS3
protease and assisting in the membrane localization of NS3 and
other viral replicase components. The formation of a NS3-NS4A
complex is necessary for proper protease activity resulting in
increased proteolytic efficiency of the cleavage events. The NS3
protein also exhibits nucleoside triphosphatase and RNA helicase
activities. NS5B (also referred to herein as HCV polymerase) is a
RNA-dependent RNA polymerase that is involved in the replication of
HCV with other HCV proteins, including NS5A, in a replicase
complex.
[0008] Compounds useful for treating HCV-infected patients are
desired which selectively inhibit HCV viral replication. In
particular, compounds which are effective to inhibit the function
of the NS5A protein are desired. The HCV NS5A protein is described,
for example, in the following references: Tan, S. L. et al.,
Virology, 284:1-12 (2001); Park, K.-J. et al., J. Biol. Chem.,
30711-30718 (2003); Tellinghuisen, T. L. et al., Nature, 435:374
(2005); Love, R. A. et al., J. Virol., 83:4395 (2009); Appel, N. et
al., J. Biol. Chem., 281:9833 (2006); Huang, L., J. Biol. Chem.,
280:36417 (2005); Rice, C. et al., World Patent Application WO
2006/093867.
[0009] In its first aspect the present disclosure provides a
compound of Formula (I)
##STR00001##
or a pharmaceutically acceptable salt thereof, wherein:
[0010] X and X' are each independently selected from CH, CR.sup.1,
and N;
[0011] Y and Y' are each independently selected from CH, CR.sup.2,
and N;
[0012] provided that no more than two of X, X', Y, and Y' are
N;
[0013] R.sup.1 and R.sup.2 are independently selected from
##STR00002## ##STR00003##
[0014] each R.sup.3 is independently selected from hydrogen, cyano,
and halo;
[0015] each R.sup.4 is independently selected from hydrogen, and
alkyl, wherein the alkyl can optionally form a fused three- to
five-membered ring with an adjacent carbon atom wherein said ring
is optionally substituted with one or two methyl groups; or,
R.sup.4 and the carbon to which it is attached form an ethylene
group;
[0016] each R.sup.5 is independently selected from hydrogen and
--C(O)R.sup.6;
[0017] each R.sup.6 is independently selected from alkoxy, alkyl,
arylalkoxy, arylalkyl, cycloalkyl, heterocyclyl, heterocyclylalkyl,
(NR.sup.cR.sup.d)alkenyl, and (NR.sup.cR.sup.d)alkyl; and
[0018] each R.sup.7 and R.sup.8 is independently selected from
hydrogen and alkyl.
[0019] In a first embodiment of the first aspect the present
disclosure provides a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, wherein:
[0020] X and X' are independently selected from CH and CR.sup.1;
and
[0021] Y, and Y' are independently selected from CH and
CR.sup.2.
[0022] In a second embodiment of the first aspect the present
disclosure provides a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, wherein:
[0023] X and X' are independently selected from CH and CR.sup.1;
and
[0024] Y and Y' are each N.
[0025] In a third embodiment of the first aspect the present
disclosure provides a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, wherein:
[0026] Y and X' are each N;
[0027] X is selected from CH and CR.sup.1; and
[0028] Y' is selected from CH and CR.sup.2.
[0029] In a fourth embodiment of the first aspect the present
disclosure provides a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, wherein:
[0030] X' is N; and
[0031] X is selected from CH and CR.sup.1; and
[0032] Y and Y' are independently selected from CH and
CR.sup.2.
[0033] In a second aspect the present disclosure provides a
composition comprising a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier. In a first embodiment of the second aspect the
composition further comprises at least one additional compound
having anti-HCV activity. In a second embodiment of the second
aspect at least one of the additional compounds is an interferon or
a ribavirin. In a third embodiment of the second aspect the
interferon is selected from interferon alpha 2B, pegylated
interferon alpha, consensus interferon, interferon alpha 2A, and
lymphoblastoid interferon tau.
[0034] In a fourth embodiment of the second aspect the present
disclosure provides a composition comprising a compound of Formula
(I), or a pharmaceutically acceptable salt thereof, a
pharmaceutically acceptable carrier, and at least one additional
compound having anti-HCV activity, wherein at least one of the
additional compounds is selected from interleukin 2, interleukin 6,
interleukin 12, a compound that enhances the development of a type
1 helper T cell response, interfering RNA, anti-sense RNA,
Imiqimod, ribavirin, an inosine 5'-monophospate dehydrogenase
inhibitor, amantadine, and rimantadine.
[0035] In a fifth embodiment of the second aspect the present
disclosure provides a composition comprising a compound of Formula
(I), or a pharmaceutically acceptable salt thereof, a
pharmaceutically acceptable carrier, and at least one additional
compound having anti-HCV activity, wherein at least one of the
additional compounds is effective to inhibit the function of a
target selected from HCV metalloprotease, HCV serine protease, HCV
polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV
assembly, HCV egress, HCV NS5A protein, and IMPDH for the treatment
of an HCV infection.
[0036] In a third aspect the present disclosure provides a method
of treating an HCV infection in a patient, comprising administering
to the patient a therapeutically effective amount of a compound of
Formula (I), or a pharmaceutically acceptable salt thereof. In a
first embodiment of the third aspect the method further comprises
administering at least one additional compound having anti-HCV
activity prior to, after or simultaneously with the compound of
Formula (I), or a pharmaceutically acceptable salt thereof. In a
second embodiment of the third aspect at least one of the
additional compounds is an interferon or a ribavirin. In a third
embodiment of the third aspect the interferon is selected from
interferon alpha 2B, pegylated interferon alpha, consensus
interferon, interferon alpha 2A, and lymphoblastoid interferon
tau.
[0037] In a fourth embodiment of the third aspect the present
disclosure provides a method of treating an HCV infection in a
patient, comprising administering to the patient a therapeutically
effective amount of a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, and at least one
additional compound having anti-HCV activity prior to, after or
simultaneously with the compound of Formula (I), or a
pharmaceutically acceptable salt thereof, wherein at least one of
the additional compounds is selected from interleukin 2,
interleukin 6, interleukin 12, a compound that enhances the
development of a type 1 helper T cell response, interfering RNA,
anti-sense RNA, Imiqimod, ribavirin, an inosine 5'-monophospate
dehydrogenase inhibitor, amantadine, and rimantadine.
[0038] In a fifth embodiment of the third aspect the present
disclosure provides a method of treating an HCV infection in a
patient, comprising administering to the patient a therapeutically
effective amount of a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, and at least one
additional compound having anti-HCV activity prior to, after or
simultaneously with the compound of Formula (I), or a
pharmaceutically acceptable salt thereof, wherein at least one of
the additional compounds is effective to inhibit the function of a
target selected from HCV metalloprotease, HCV serine protease, HCV
polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV
assembly, HCV egress, HCV NS5A protein, and IMPDH for the treatment
of an HCV infection.
[0039] Other aspects of the present disclosure may include suitable
combinations of embodiments disclosed herein.
[0040] Yet other aspects and embodiments may be found in the
description provided herein.
[0041] The description of the present disclosure herein should be
construed in congruity with the laws and principals of chemical
bonding. In some instances it may be necessary to remove a hydrogen
atom in order accommodate a substituent at any given location.
[0042] It should be understood that the compounds encompassed by
the present disclosure are those that are suitably stable for use
as pharmaceutical agent.
[0043] It is intended that the definition of any substituent or
variable at a particular location in a molecule be independent of
its definitions elsewhere in that molecule. For example, when
R.sup.1 and R.sup.2 both contain an R.sup.4 group, the two R.sup.4
groups may be the same or different.
[0044] All patents, patent applications, and literature references
cited in the specification are herein incorporated by reference in
their entirety. In the case of inconsistencies, the present
disclosure, including definitions, will prevail.
[0045] As used in the present specification, the following terms
have the meanings indicated:
[0046] As used herein, the singular forms "a", "an", and "the"
include plural reference unless the context clearly dictates
otherwise.
[0047] Unless stated otherwise, all aryl, cycloalkyl, and
heterocyclyl groups of the present disclosure may be substituted as
described in each of their respective definitions. For example, the
aryl part of an arylalkyl group may be substituted as described in
the definition of the term "aryl".
[0048] The term "alkenyl," as used herein, refers to a straight or
branched chain group of two to six carbon atoms containing at least
one carbon-carbon double bond.
[0049] The term "alkenyloxy," as used herein, refers to an alkenyl
group attached to the parent molecular moiety through an oxygen
atom.
[0050] The term "alkenyloxycarbonyl," as used herein, refers to an
alkenyloxy group attached to the parent molecular moiety through a
carbonyl group.
[0051] The term "alkoxy," as used herein, refers to an alkyl group
attached to the parent molecular moiety through an oxygen atom.
[0052] The term "alkoxyalkyl," as used herein, refers to an alkyl
group substituted with one, two, or three alkoxy groups.
[0053] The term "alkoxyalkylcarbonyl," as used herein, refers to an
alkoxyalkyl group attached to the parent molecular moiety through a
carbonyl group.
[0054] The term "alkoxycarbonyl," as used herein, refers to an
alkoxy group attached to the parent molecular moiety through a
carbonyl group.
[0055] The term "alkylcarbonyloxy," as used herein, refers to an
alkylcarbonyl group attached to the parent molecular moiety through
an oxygen atom.
[0056] The term "alkyl," as used herein, refers to a group derived
from a straight or branched chain saturated hydrocarbon containing
from one to six carbon atoms. In the compounds of the present
disclosure, when R.sup.4 is alkyl, the alkyl can optionally form a
fused three-membered ring with an adjacent carbon atom to provide
the structure shown below.
##STR00004##
wherein n is selected from 1 and 2.
[0057] The term "alkylcarbonyl," as used herein, refers to an alkyl
group attached to the parent molecular moiety through a carbonyl
group.
[0058] The term "alkylsulfanyl," as used herein, refers to an alkyl
group attached to the parent molecular moiety through a sulfur
atom.
[0059] The term "alkylsulfonyl," as used herein, refers to an alkyl
group attached to the parent molecular moiety through a sulfonyl
group.
[0060] The term "aryl," as used herein, refers to a phenyl group,
or a bicyclic fused ring system wherein one or both of the rings is
a phenyl group. Bicyclic fused ring systems consist of a phenyl
group fused to a four- to six-membered aromatic or non-aromatic
carbocyclic ring. The aryl groups of the present disclosure can be
attached to the parent molecular moiety through any substitutable
carbon atom in the group. Representative examples of aryl groups
include, but are not limited to, indanyl, indenyl, naphthyl,
phenyl, and tetrahydronaphthyl. The aryl groups of the present
disclosure are optionally substituted with one, two, three, four,
or five substituents independently selected from alkenyl, alkoxy,
alkoxyalkyl, alkoxycarbonyl, alkyl, alkylcarbonyl, a second aryl
group, arylalkoxy, arylalkyl, arylcarbonyl, cyano, halo,
haloalkoxy, haloalkyl, heterocyclyl, heterocyclylalkyl,
heterocyclylcarbonyl, hydroxy, hydroxyalkyl, nitro,
--NR.sup.xR.sup.y, (NR.sup.xR.sup.y)alkyl, oxo, and --P(O)OR.sub.2,
wherein each R is independently selected from hydrogen and alkyl;
and wherein the alkyl part of the arylalkyl and the
heterocyclylalkyl are unsubstituted and wherein the second aryl
group, the aryl part of the arylalkyl, the aryl part of the
arylcarbonyl, the heterocyclyl, and the heterocyclyl part of the
heterocyclylalkyl and the heterocyclylcarbonyl are further
optionally substituted with one, two, or three substituents
independently selected from alkoxy, alkyl, cyano, halo, haloalkoxy,
haloalkyl, and nitro.
[0061] The term "arylalkoxy," as used herein, refers to an aryl
group attached to the parent molecular moiety through an alkoxy
group.
[0062] The term "arylalkoxycarbonyl," as used herein, refers to an
arylalkoxy group attached to the parent molecular moiety through a
carbonyl group.
[0063] The term "arylalkyl," as used herein, refers to an alkyl
group substituted with one, two, or three aryl groups. The alkyl
part of the arylalkyl is further optionally substituted with one or
two additional groups independently selected from alkoxy,
alkylcarbonyloxy, halo, haloalkoxy, haloalkyl, heterocyclyl,
hydroxy, and --NR.sup.cR.sup.d, wherein the heterocyclyl is further
optionally substituted with one or two substituents independently
selected from alkoxy, alkyl, unsubstituted aryl, unsubstituted
arylalkoxy, unsubstituted arylalkoxycarbonyl, halo, haloalkoxy,
haloalkyl, hydroxy, --NR.sup.xR.sup.y, and oxo.
[0064] The term "arylcarbonyl," as used herein, refers to an aryl
group attached to the parent molecular moiety through a carbonyl
group.
[0065] The term "cyano," as used herein, refers to --CN.
[0066] The term "cycloalkyl," as used herein, refers to a saturated
monocyclic or bicyclic hydrocarbon ring system having three to
fourteen carbon atoms and zero heteroatoms. Representative examples
of cycloalkyl groups include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[3.1.1]heptyl, and
adamantyl. The cycloalkyl groups of the present disclosure are
optionally substituted with one, two, three, four, or five
substituents independently selected from alkoxy, alkyl, aryl,
cyano, halo, haloalkoxy, haloalkyl, heterocyclyl, hydroxy,
hydroxyalkyl, nitro, and --NR.sup.xR.sup.y, wherein the aryl and
the heterocyclyl are further optionally substituted with one, two,
or three substituents independently selected from alkoxy, alkyl,
cyano, halo, haloalkoxy, haloalkyl, hydroxy, nitro, and oxo.
[0067] The term "cycloalkyloxy," as used herein, refers to a
cycloalkyl group attached to the parent molecular moiety through an
oxygen atom.
[0068] The term "cycloalkyloxycarbonyl," as used herein, refers to
a cycloalkyloxy group attached to the parent molecular moiety
through a carbonyl group.
[0069] The term "cycloalkylsulfonyl," as used herein, refers to a
cycloalkyl group attached to the parent molecular moiety through a
sulfonyl group.
[0070] The term "ethylene," as used herein, refers to
.dbd.CH.sub.2.
[0071] The term "formyl," as used herein, refers to --CHO.
[0072] The terms "halo" and "halogen," as used herein, refers to
Cl, Br, F, or I.
[0073] The term "haloalkoxy," as used herein, refers to a haloalkyl
group attached to the parent molecular moiety through an oxygen
atom.
[0074] The term "haloalkoxycarbonyl," as used herein, refers to a
haloalkoxy group attached to the parent molecular moiety through a
carbonyl group.
[0075] The term "haloalkyl," as used herein, refers to an alkyl
group substituted with one, two, three, or four halogen atoms.
[0076] The term "heterocyclyl," as used herein, refers to a four-,
five-, six-, or seven-membered ring containing one, two, three, or
four heteroatoms independently selected from nitrogen, oxygen, and
sulfur. The four-membered ring has zero double bonds, the
five-membered ring has zero to two double bonds, and the six- and
seven-membered rings have zero to three double bonds. The term
"heterocyclyl" also includes bicyclic groups in which the
heterocyclyl ring is fused to another monocyclic heterocyclyl group
or a three- to seven-membered aromatic or non-aromatic carbocyclic
ring; bicyclic groups in which the heterocyclyl ring is substituted
with a three- to seven-membered spirocyclic ring; as well as
bridged bicyclic groups such as 7-azabicyclo[2.2.1]hept-7-yl,
2-azabicyclo[2.2.2]oct-2-yl, and 2-azabicyclo[2.2.2]oct-3-yl. The
heterocyclyl groups of the present disclosure can be attached to
the parent molecular moiety through any carbon atom or nitrogen
atom in the group. Examples of heterocyclyl groups include, but are
not limited to, benzothienyl, furyl, imidazolyl, indolinyl,
indolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl,
oxazolyl, piperazinyl, piperidinyl, pyrazolyl, pyridinyl,
pyrrolidinyl, pyrrolopyridinyl, pyrrolyl, quinolinyl,
tetrahydropyranyl, thiazolyl, thienyl, and thiomorpholinyl. The
heterocyclyl groups of the present disclosure are optionally
substituted with one, two, three, four, or five substituents
independently selected from alkenyl, alkoxy, alkoxyalkyl,
alkoxycarbonyl, alkyl, alkylcarbonyl, aryl, arylalkoxycarbonyl,
arylalkyl, arylcarbonyl, cyano, halo, haloalkoxy, haloalkyl, a
second heterocyclyl group, heterocyclylalkyl, heterocyclylcarbonyl,
hydroxy, hydroxyalkyl, nitro, --NR.sup.xR.sup.y,
(NR.sup.xR.sup.y)alkyl, and oxo, wherein the alkyl part of the
arylalkyl and the heterocyclylalkyl are unsubstituted and wherein
the aryl, the aryl part of the arylalkyl, the aryl part of the
arylcarbonyl, the second heterocyclyl group, and the heterocyclyl
part of the heterocyclylalkyl and the heterocyclylcarbonyl are
further optionally substituted with one, two, or three substituents
independently selected from alkoxy, alkyl, cyano, halo, haloalkoxy,
haloalkyl, and nitro.
[0077] The term "heterocyclylalkoxy," as used herein, refers to a
heterocyclyl group attached to the parent molecular moiety through
an alkoxy group.
[0078] The term "heterocyclylalkoxycarbonyl," as used herein,
refers to a heterocyclylalkoxy group attached to the parent
molecular moiety through a carbonyl group.
[0079] The term "heterocyclylalkyl," as used herein, refers to an
alkyl group substituted with one, two, or three heterocyclyl
groups. The alkyl part of the heterocyclylalkyl is further
optionally substituted with one or two additional groups
independently selected from alkoxy, alkylcarbonyloxy, aryl, halo,
haloalkoxy, haloalkyl, hydroxy, and --NR.sup.cR.sup.d, wherein the
aryl is further optionally substituted with one or two substituents
independently selected from alkoxy, alkyl, unsubstituted aryl,
unsubstituted arylalkoxy, unsubstituted arylalkoxycarbonyl, halo,
haloalkoxy, haloalkyl, hydroxy, and --NR.sup.xR.sup.y.
[0080] The term "heterocyclylalkylcarbonyl," as used herein, refers
to a heterocyclylalkyl group attached to the parent molecular
moiety through a carbonyl group.
[0081] The term "heterocyclylcarbonyl," as used herein, refers to a
heterocyclyl group attached to the parent molecular moiety through
a carbonyl group.
[0082] The term "heterocyclyloxy," as used herein, refers to a
heterocyclyl group attached to the parent molecular moiety through
an oxygen atom.
[0083] The term "heterocyclyloxycarbonyl," as used herein, refers
to a heterocyclyloxy group attached to the parent molecular moiety
through a carbonyl group.
[0084] The term "hydroxy," as used herein, refers to --OH.
[0085] The term "hydroxyalkyl," as used herein, refers to an alkyl
group substituted with one, two, or three hydroxy groups.
[0086] The term "hydroxyalkylcarbonyl," as used herein, refers to a
hydroxyalkyl group attached to the parent molecular moiety through
a carbonyl group.
[0087] The term "nitro," as used herein, refers to --NO.sub.2.
[0088] The term "--NR.sup.cR.sup.d," as used herein, refers to two
groups, R.sup.c and R.sup.d, which are attached to the parent
molecular moiety through a nitrogen atom. R.sup.c and R.sup.d are
independently selected from hydrogen, alkenyloxycarbonyl,
alkoxyalkylcarbonyl, alkoxycarbonyl, alkyl, alkylcarbonyl,
cycloalkyloxy, alkylsulfonyl, aryl, arylalkoxycarbonyl, arylalkyl,
arylalkylcarbonyl, arylcarbonyl, aryloxycarbonyl, arylsulfonyl,
cycloalkyl, cycloalkyloxycarbonyl, cycloalkylsulfonyl, formyl,
haloalkoxycarbonyl, heterocyclyl, heterocyclylalkoxycarbonyl,
heterocyclylalkyl, heterocyclylalkylcarbonyl, heterocyclylcarbonyl,
heterocyclyloxycarbonyl, hydroxyalkylcarbonyl,
(NR.sup.eR.sup.f)alkyl, (NR.sup.eR.sup.f)alkylcarbonyl,
(NR.sup.eR.sup.f)carbonyl, (NR.sup.eR.sup.f)sulfonyl, --C(NCN)OR',
and --C(NCN)NR.sup.xR.sup.y, wherein R' is selected from alkyl and
unsubstituted phenyl, and wherein the alkyl part of the arylalkyl,
the arylalkylcarbonyl, the heterocyclylalkyl, and the
heterocyclylalkylcarbonyl are further optionally substituted with
one --NR.sup.eR.sup.f group; and wherein the aryl, the aryl part of
the arylalkoxycarbonyl, the arylalkyl, the arylalkylcarbonyl, the
arylcarbonyl, the aryloxycarbonyl, and the arylsulfonyl, the
heterocyclyl, and the heterocyclyl part of the
heterocyclylalkoxycarbonyl, the heterocyclylalkyl, the
heterocyclylalkylcarbonyl, the heterocyclylcarbonyl, and the
heterocyclyloxycarbonyl are further optionally substituted with
one, two, or three substituents independently selected from alkoxy,
alkyl, cyano, halo, haloalkoxy, haloalkyl, and nitro.
[0089] The term "(NR.sup.cR.sup.d)alkenyl," as used herein, refers
to
##STR00005##
wherein R.sup.c and R.sup.d are as defined herein and each R.sup.q
is independently hydrogen or C.sub.1-3 alkyl.
[0090] The term "(NR.sup.cR.sup.d)alkyl," as used herein, refers to
an alkyl group substituted with one, two, or three
--NR.sup.cR.sup.d groups. The alkyl part of the
(NR.sup.cR.sup.d)alkyl is further optionally substituted with one
or two additional groups selected from alkoxy, alkoxyalkylcarbonyl,
alkoxycarbonyl, alkylsulfanyl, C.sub.2 alkynyl, arylalkoxycarbonyl,
carboxy, cyano, cycloalkyl, halo, heterocyclyl,
heterocyclylcarbonyl, hydroxy, and (NR.sup.eR.sup.f)carbonyl;
wherein the heterocyclyl is further optionally substituted with
one, two, three, four, or five substituents independently selected
from alkoxy, alkyl, cyano, halo, haloalkoxy, haloalkyl, and
nitro.
[0091] The term "--NR.sup.eR.sup.f," as used herein, refers to two
groups, R.sup.e and R.sup.f, which are attached to the parent
molecular moiety through a nitrogen atom. R.sup.e and R.sup.f are
independently selected from hydrogen, alkyl, unsubstituted aryl,
unsubstituted arylalkyl, unsubstituted cycloalkyl, unsubstituted
(cyclolalkyl)alkyl, unsubstituted heterocyclyl, unsubstituted
heterocyclylalkyl, (NR.sup.xR.sup.y)alkyl, and
(NR.sup.xR.sup.y)carbonyl.
[0092] The term "(NR.sup.eR.sup.f)alkyl," as used herein, refers to
an alkyl group substituted with one, two, or three
--NR.sup.eR.sup.f groups.
[0093] The term "(NR.sup.eR.sup.f)alkylcarbonyl," as used herein,
refers to an (NR.sup.eR.sup.f)alkyl group attached to the parent
molecular moiety through a carbonyl group.
[0094] The term "(NR.sup.eR.sup.f)carbonyl," as used herein, refers
to an --NR.sup.eR.sup.f group attached to the parent molecular
moiety through a carbonyl group.
[0095] The term "(NR.sup.eR.sup.f)sulfonyl," as used herein, refers
to an --NR.sup.eR.sup.f group attached to the parent molecular
moiety through a sulfonyl group.
[0096] The term "--NR.sup.xR.sup.y," as used herein, refers to two
groups, R.sup.x and R.sup.y, which are attached to the parent
molecular moiety through a nitrogen atom. R.sup.x and R.sup.y are
independently selected from hydrogen, alkoxycarbonyl, alkyl,
alkylcarbonyl, unsubstituted aryl, unsubstituted
arylalkoxycarbonyl, unsubstituted arylalkyl, unsubstituted
cycloalkyl, unsubstituted heterocyclyl, and
(NR.sup.x'R.sup.y')carbonyl, wherein R.sup.x' and R.sup.y' are
independently selected from hydrogen and alkyl.
[0097] The term "(NR.sup.xR.sup.y)alkyl," as used herein, refers to
an alkyl group substituted with one, two, or three
--NR.sup.xR.sup.y groups.
[0098] The term "(NR.sup.xR.sup.y)carbonyl," as used herein, refers
to an --NR.sup.xR.sup.y group attached to the parent molecular
moiety through a carbonyl group.
[0099] The term "--NR.sup.xR.sup.y," as used herein, refers to two
groups, R.sup.x and R.sup.y, which are attached to the parent
molecular moiety through a nitrogen atom. R.sup.x and R.sup.y are
independently selected from hydrogen and alkyl.
[0100] The term "(NR.sup.x'R.sup.y')carbonyl," as used herein,
refers to an --NR.sup.x'R.sup.y' group attached to the parent
molecular moiety through a carbonyl group.
[0101] The term "oxo," as used herein, refers to .dbd.O.
[0102] The term "sulfonyl," as used herein, refers to
--SO.sub.2--.
[0103] Asymmetric centers exist in the compounds of the present
disclosure. These centers are designated by the symbols "R" or "S",
depending on the configuration of substituents around the chiral
carbon atom. It should be understood that the disclosure
encompasses all stereochemical isomeric forms, or mixtures thereof,
which possess the ability to inhibit NS5A. Individual stereoisomers
of compounds can be prepared synthetically from commercially
available starting materials which contain chiral centers or by
preparation of mixtures of stereoisomeric products followed by
separation such as conversion to a mixture of diastereomers
followed by separation or recrystallization, chromatographic
techniques, or direct separation of stereoisomers on chiral
chromatographic columns. Starting compounds of particular
stereochemistry are either commercially available or can be made
and resolved by techniques known in the art.
[0104] Certain compounds of the present disclosure may also exist
in different stable conformational forms which may be separable.
Torsional asymmetry due to restricted rotation about an asymmetric
single bond, for example because of steric hindrance or ring
strain, may permit separation of different conformers. The present
disclosure includes each conformational isomer of these compounds
and mixtures thereof.
[0105] The compounds of the present disclosure also exist as
tautomers; therefore the present disclosure also encompasses all
tautomeric forms.
[0106] The term "compounds of the present disclosure", and
equivalent expressions, are meant to embrace compounds of Formula
(I), and pharmaceutically acceptable stereoisomers, diastereomers,
and salts thereof. Similarly, references to intermediates are meant
to embrace their salts where the context so permits.
[0107] The present disclosure is intended to include all isotopes
of atoms occurring in the present compounds. Isotopes include those
atoms having the same atomic number but different mass numbers. By
way of general example and without limitation, isotopes of hydrogen
include deuterium and tritium. Isotopes of carbon include .sup.13C
and .sup.14C. Isotopically-labeled compounds of the invention can
generally be prepared by conventional techniques known to those
skilled in the art or by processes analogous to those described
herein, using an appropriate isotopically-labeled reagent in place
of the non-labeled reagent otherwise employed. Such compounds may
have a variety of potential uses, for example as standards and
reagents in determining biological activity. In the case of stable
isotopes, such compounds may have the potential to favorably modify
biological, pharmacological, or pharmacokinetic properties.
[0108] The compounds of the present disclosure can exist as
pharmaceutically acceptable salts. The term "pharmaceutically
acceptable salt," as used herein, represents salts or zwitterionic
forms of the compounds of the present disclosure which are water or
oil-soluble or dispersible, which are, within the scope of sound
medical judgment, suitable for use in contact with the tissues of
patients without excessive toxicity, irritation, allergic response,
or other problem or complication commensurate with a reasonable
benefit/risk ratio, and are effective for their intended use. The
salts can be prepared during the final isolation and purification
of the compounds or separately by reacting a suitable nitrogen atom
with a suitable acid. Representative acid addition salts include
acetate, adipate, alginate, citrate, aspartate, benzoate,
benzenesulfonate, bisulfate, butyrate, camphorate,
camphorsulfonate; digluconate, glycerophosphate, hemisulfate,
heptanoate, hexanoate, formate, fumarate, hydrochloride,
hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate,
maleate, mesitylenesulfonate, methanesulfonate,
naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate,
palmoate, pectinate, persulfate, 3-phenylproprionate, picrate,
pivalate, propionate, succinate, tartrate, trichloroacetate,
trifluoroacetate, phosphate, glutamate, bicarbonate,
para-toluenesulfonate, and undecanoate. Examples of acids which can
be employed to form pharmaceutically acceptable addition salts
include inorganic acids such as hydrochloric, hydrobromic,
sulfuric, and phosphoric, and organic acids such as oxalic, maleic,
succinic, and citric.
[0109] Basic addition salts can be prepared during the final
isolation and purification of the compounds by reacting a carboxy
group with a suitable base such as the hydroxide, carbonate, or
bicarbonate of a metal cation or with ammonia or an organic
primary, secondary, or tertiary amine. The cations of
pharmaceutically acceptable salts include lithium, sodium,
potassium, calcium, magnesium, and aluminum, as well as nontoxic
amine cations such as ammonium, tetramethylammonium,
tetraethylammonium, methylamine, dimethylamine, trimethylamine,
triethylamine, diethylamine, ethylamine, tributylamine, pyridine,
N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,
dicyclohexylamine, procaine, dibenzylamine,
N,N-dibenzylphenethylamine, and N,N'-dibenzylethylenediamine. Other
representative organic amines useful for the formation of base
addition salts include ethylenediamine, ethanolamine,
diethanolamine, piperidine, and piperazine.
[0110] When it is possible that, for use in therapy,
therapeutically effective amounts of a compound of Formula (I), as
well as pharmaceutically acceptable salts thereof, may be
administered as the raw chemical, it is possible to present the
active ingredient as a pharmaceutical composition. Accordingly, the
disclosure further provides pharmaceutical compositions, which
include therapeutically effective amounts of compounds of Formula
(I) or pharmaceutically acceptable salts thereof, and one or more
pharmaceutically acceptable carriers, diluents, or excipients. The
term "therapeutically effective amount," as used herein, refers to
the total amount of each active component that is sufficient to
show a meaningful patient benefit, e.g., a sustained reduction in
viral load. When applied to an individual active ingredient,
administered alone, the term refers to that ingredient alone. When
applied to a combination, the term refers to combined amounts of
the active ingredients that result in the therapeutic effect,
whether administered in combination, serially, or simultaneously.
The compounds of Formula (I) and pharmaceutically acceptable salts
thereof, are as described above. The carrier(s), diluent(s), or
excipient(s) must be acceptable in the sense of being compatible
with the other ingredients of the formulation and not deleterious
to the recipient thereof. In accordance with another aspect of the
present disclosure there is also provided a process for the
preparation of a pharmaceutical formulation including admixing a
compound of Formula (I), or a pharmaceutically acceptable salt
thereof, with one or more pharmaceutically acceptable carriers,
diluents, or excipients. The term "pharmaceutically acceptable," as
used herein, refers to those compounds, materials, compositions,
and/or dosage forms which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of patients
without excessive toxicity, irritation, allergic response, or other
problem or complication commensurate with a reasonable benefit/risk
ratio, and are effective for their intended use.
[0111] Pharmaceutical formulations may be presented in unit dose
forms containing a predetermined amount of active ingredient per
unit dose. Dosage levels of between about 0.01 and about 250
milligram per kilogram ("mg/kg") body weight per day, preferably
between about 0.05 and about 100 mg/kg body weight per day of the
compounds of the present disclosure are typical in a monotherapy
for the prevention and treatment of HCV mediated disease.
Typically, the pharmaceutical compositions of this disclosure will
be administered from about 1 to about 5 times per day or
alternatively, as a continuous infusion. Such administration can be
used as a chronic or acute therapy. The amount of active ingredient
that may be combined with the carrier materials to produce a single
dosage form will vary depending on the condition being treated, the
severity of the condition, the time of administration, the route of
administration, the rate of excretion of the compound employed, the
duration of treatment, and the age, gender, weight, and condition
of the patient. Preferred unit dosage formulations are those
containing a daily dose or sub-dose, as herein above recited, or an
appropriate fraction thereof, of an active ingredient. Generally,
treatment is initiated with small dosages substantially less than
the optimum dose of the compound. Thereafter, the dosage is
increased by small increments until the optimum effect under the
circumstances is reached. In general, the compound is most
desirably administered at a concentration level that will generally
afford antivirally effective results without causing any harmful or
deleterious side effects.
[0112] When the compositions of this disclosure comprise a
combination of a compound of the present disclosure and one or more
additional therapeutic or prophylactic agent, both the compound and
the additional agent are usually present at dosage levels of
between about 10 to 150%, and more preferably between about 10 and
80% of the dosage normally administered in a monotherapy
regimen.
[0113] Pharmaceutical formulations may be adapted for
administration by any appropriate route, for example by the oral
(including buccal or sublingual), rectal, nasal, topical (including
buccal, sublingual, or transdermal), vaginal, or parenteral
(including subcutaneous, intracutaneous, intramuscular,
intra-articular, intrasynovial, intrasternal, intrathecal,
intralesional, intravenous, or intradermal injections or infusions)
route. Such formulations may be prepared by any method known in the
art of pharmacy, for example by bringing into association the
active ingredient with the carrier(s) or excipient(s). Oral
administration or administration by injection are preferred.
[0114] Pharmaceutical formulations adapted for oral administration
may be presented as discrete units such as capsules or tablets;
powders or granules; solutions or suspensions in aqueous or
non-aqueous liquids; edible foams or whips; or oil-in-water liquid
emulsions or water-in-oil emulsions.
[0115] For instance, for oral administration in the form of a
tablet or capsule, the active drug component can be combined with
an oral, non-toxic pharmaceutically acceptable inert carrier such
as ethanol, glycerol, water, and the like. Powders are prepared by
comminuting the compound to a suitable fine size and mixing with a
similarly comminuted pharmaceutical carrier such as an edible
carbohydrate, as, for example, starch or mannitol. Flavoring,
preservative, dispersing, and coloring agent can also be
present.
[0116] Capsules are made by preparing a powder mixture, as
described above, and filling formed gelatin sheaths. Glidants and
lubricants such as colloidal silica, talc, magnesium stearate,
calcium stearate, or solid polyethylene glycol can be added to the
powder mixture before the filling operation. A disintegrating or
solubilizing agent such as agar-agar, calcium carbonate, or sodium
carbonate can also be added to improve the availability of the
medicament when the capsule is ingested.
[0117] Moreover, when desired or necessary, suitable binders,
lubricants, disintegrating agents, and coloring agents can also be
incorporated into the mixture. Suitable binders include starch,
gelatin, natural sugars such as glucose or beta-lactose, corn
sweeteners, natural and synthetic gums such as acacia, tragacanth
or sodium alginate, carboxymethylcellulose, polyethylene glycol,
and the like. Lubricants used in these dosage forms include sodium
oleate, sodium chloride, and the like. Disintegrators include,
without limitation, starch, methyl cellulose, agar, betonite,
xanthan gum, and the like. Tablets are formulated, for example, by
preparing a powder mixture, granulating or slugging, adding a
lubricant and disintegrant, and pressing into tablets. A powder
mixture is prepared by mixing the compound, suitable comminuted,
with a diluent or base as described above, and optionally, with a
binder such as carboxymethylcellulose, an aliginate, gelating, or
polyvinyl pyrrolidone, a solution retardant such as paraffin, a
resorption accelerator such as a quaternary salt and/or and
absorption agent such as betonite, kaolin, or dicalcium phosphate.
The powder mixture can be granulated by wetting with a binder such
as syrup, starch paste, acadia mucilage, or solutions of cellulosic
or polymeric materials and forcing through a screen. As an
alternative to granulating, the powder mixture can be run through
the tablet machine and the result is imperfectly formed slugs
broken into granules. The granules can be lubricated to prevent
sticking to the tablet forming dies by means of the addition of
stearic acid, a stearate salt, talc, or mineral oil. The lubricated
mixture is then compressed into tablets. The compounds of the
present disclosure can also be combined with a free flowing inert
carrier and compressed into tablets directly without going through
the granulating or slugging steps. A clear or opaque protective
coating consisting of a sealing coat of shellac, a coating of sugar
or polymeric material, and a polish coating of wax can be provided.
Dyestuffs can be added to these coatings to distinguish different
unit dosages.
[0118] Oral fluids such as solution, syrups, and elixirs can be
prepared in dosage unit form so that a given quantity contains a
predetermined amount of the compound. Syrups can be prepared by
dissolving the compound in a suitably flavored aqueous solution,
while elixirs are prepared through the use of a non-toxic vehicle.
Solubilizers and emulsifiers such as ethoxylated isostearyl
alcohols and polyoxyethylene sorbitol ethers, preservatives, flavor
additive such as peppermint oil or natural sweeteners, or saccharin
or other artificial sweeteners, and the like can also be added.
[0119] Where appropriate, dosage unit formulations for oral
administration can be microencapsulated. The formulation can also
be prepared to prolong or sustain the release as for example by
coating or embedding particulate material in polymers, wax, or the
like.
[0120] The compounds of Formula (I), and pharmaceutically
acceptable salts thereof, can also be administered in the form of
liposome delivery systems, such as small unilamellar vesicles,
large unilamellar vesicles, and multilamellar vesicles. Liposomes
can be formed from a variety of phopholipids, such as cholesterol,
stearylamine, or phophatidylcholines.
[0121] The compounds of Formula (I) and pharmaceutically acceptable
salts thereof may also be delivered by the use of monoclonal
antibodies as individual carriers to which the compound molecules
are coupled. The compounds may also be coupled with soluble
polymers as targetable drug carriers. Such polymers can include
polyvinylpyrrolidone, pyran copolymer,
polyhydroxypropylmethacrylamidephenol,
polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine
substituted with palitoyl residues. Furthermore, the compounds may
be coupled to a class of biodegradable polymers useful in achieving
controlled release of a drug, for example, polylactic acid,
polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters,
polyacetals, polydihydropyrans, polycyanoacrylates, and
cross-linked or amphipathic block copolymers of hydrogels.
[0122] Pharmaceutical formulations adapted for transdermal
administration may be presented as discrete patches intended to
remain in intimate contact with the epidermis of the recipient for
a prolonged period of time. For example, the active ingredient may
be delivered from the patch by iontophoresis as generally described
in Pharmaceutical Research, 3(6):318 (1986).
[0123] Pharmaceutical formulations adapted for topical
administration may be formulated as ointments, creams, suspensions,
lotions, powders, solutions, pastes, gels, sprays, aerosols, or
oils.
[0124] For treatments of the eye or other external tissues, for
example mouth and skin, the formulations are preferably applied as
a topical ointment or cream. When formulated in an ointment, the
active ingredient may be employed with either a paraffinic or a
water-miscible ointment base. Alternatively, the active ingredient
may be formulated in a cream with an oil-in-water cream base or a
water-in oil base.
[0125] Pharmaceutical formulations adapted for topical
administrations to the eye include eye drops wherein the active
ingredient is dissolved or suspended in a suitable carrier,
especially an aqueous solvent.
[0126] Pharmaceutical formulations adapted for topical
administration in the mouth include lozenges, pastilles, and mouth
washes.
[0127] Pharmaceutical formulations adapted for rectal
administration may be presented as suppositories or as enemas.
[0128] Pharmaceutical formulations adapted for nasal administration
wherein the carrier is a solid include a course powder having a
particle size for example in the range 20 to 500 microns which is
administered in the manner in which snuff is taken, i.e., by rapid
inhalation through the nasal passage from a container of the powder
held close up to the nose. Suitable formulations wherein the
carrier is a liquid, for administration as a nasal spray or nasal
drops, include aqueous or oil solutions of the active
ingredient.
[0129] Pharmaceutical formulations adapted for administration by
inhalation include fine particle dusts or mists, which may be
generated by means of various types of metered, dose pressurized
aerosols, nebulizers, or insufflators.
[0130] Pharmaceutical formulations adapted for vaginal
administration may be presented as pessaries, tampons, creams,
gels, pastes, foams, or spray formulations.
[0131] Pharmaceutical formulations adapted for parenteral
administration include aqueous and non-aqueous sterile injection
solutions which may contain anti-oxidants, buffers, bacteriostats,
and soutes which render the formulation isotonic with the blood of
the intended recipient; and aqueous and non-aqueous sterile
suspensions which may include suspending agents and thickening
agents. The formulations may be presented in unit-dose or
multi-dose containers, for example sealed ampoules and vials, and
may be stored in a freeze-dried (lyophilized) condition requiring
only the addition of the sterile liquid carrier, for example water
for injections, immediately prior to use.
[0132] Extemporaneous injection solutions and suspensions may be
prepared from sterile powders, granules, and tablets.
[0133] It should be understood that in addition to the ingredients
particularly mentioned above, the formulations may include other
agents conventional in the art having regard to the type of
formulation in question, for example those suitable for oral
administration may include flavoring agents.
[0134] The term "patient" includes both human and other
mammals.
[0135] The term "treating" refers to: (i) preventing a disease,
disorder or condition from occurring in a patient that may be
predisposed to the disease, disorder, and/or condition but has not
yet been diagnosed as having it; (ii) inhibiting the disease,
disorder, or condition, i.e., arresting its development; and (iii)
relieving the disease, disorder, or condition, i.e., causing
regression of the disease, disorder, and/or condition.
[0136] The compounds of the present disclosure can also be
administered with a cyclosporin, for example, cyclosporin A.
Cyclosporin A has been shown to be active against HCV in clinical
trials (Hepatology, 38:1282 (2003); Biochem. Biophys. Res. Commun.,
313:42 (2004); J. Gastroenterol., 38:567 (2003)).
[0137] Table 1 below lists some illustrative examples of compounds
that can be administered with the compounds of this disclosure. The
compounds of the disclosure can be administered with other anti-HCV
activity compounds in combination therapy, either jointly or
separately, or by combining the compounds into a composition.
TABLE-US-00001 TABLE 1 Type of Inhibitor or Brand Name
Physiological Class Target Source Company NIM811 Cyclophilin
Novartis Debio-025 inhibitors Debiopharm Zadaxin Immunomodulator
Sciclone Suvus Methylene blue Bioenvision Actilon (CPG10101) TLR9
agonist Coley Batabulin (T67) Anticancer .beta.-Tubulin inhibitor
Tularik Inc., South San Francisco, CA ISIS 14803 Antiviral
Antisense ISIS Pharmaceuticals Inc, Carlsbad, CA/ Elan
Pharmaceuticals Inc., New York, NY Summetrel Antiviral Antiviral
Endo Pharmaceuticals Holdings Inc., Chadds Ford, PA GS-9132
(ACH-806) Antiviral HCV inhibitor Achillion/Gilead
Pyrazolopyrimidine Antiviral HCV inhibitors Arrow Therapeutics
compounds and salts Ltd. From WO 2005/047288 26 May 2005 Levovirin
Antiviral IMPDH inhibitor Ribapharm Inc., Costa Mesa, CA
Merimepodib Antiviral IMPDH inhibitor Vertex (VX-497)
Pharmaceuticals Inc., Cambridge, MA XTL-6865 (XTL-002) Antiviral
Monoclonal XTL antibody Biopharmaceuticals Ltd., Rehovot, Israel
Telaprevir Antiviral NS3 serine Vertex (VX-950, LY-570310) protease
inhibitor Pharmaceuticals Inc., Cambridge, MA/Eli Lilly and Co.
Inc., Indianapolis, IN HCV-796 Antiviral NS5B replicase
Wyeth/Viropharma inhibitor NM-283 Antiviral NS5B replicase
Idenix/Novartis inhibitor GL-59728 Antiviral NS5B replicase Gene
Labs/Novartis inhibitor GL-60667 Antiviral NS5B replicase Gene
Labs/Novartis inhibitor 2'C MeA Antiviral NS5B replicase Gilead
inhibitor PSI 6130 Antiviral NS5B replicase Roche inhibitor R1626
Antiviral NS5B replicase Roche inhibitor 2'C Methyl adenosine
Antiviral NS5B replicase Merck inhibitor JTK-003 Antiviral RdRp
inhibitor Japan Tobacco Inc., Tokyo, Japan Levovirin Antiviral
Ribavirin ICN Pharmaceuticals, Costa Mesa, CA Ribavirin Antiviral
Ribavirin Schering-Plough Corporation, Kenilworth, NJ Viramidine
Antiviral Ribavirin prodrug Ribapharm Inc., Costa Mesa, CA
Heptazyme Antiviral Ribozyme Ribozyme Pharmaceuticals Inc.,
Boulder, CO BILN-2061 Antiviral Serine protease Boehringer
Ingelheim inhibitor Pharma KG, Ingelheim, Germany SCH 503034
Antiviral Serine protease Schering-Plough inhibitor Zadazim Immune
modulator Immune modulator SciClone Pharmaceuticals Inc., San
Mateo, CA Ceplene Immunomodulator Immune modulator Maxim
Pharmaceuticals Inc., San Diego, CA CELLCEPT .RTM.
Immunosuppressant HCV IgG immuno- F. Hoffmann-La suppressant Roche
LTD, Basel, Switzerland Civacir Immunosuppressant HCV IgG immuno-
Nabi suppressant Biopharmaceuticals Inc., Boca Raton, FL Albuferon
- .alpha. Interferon Albumin IFN-.alpha.2b Human Genome Sciences
Inc., Rockville, MD Infergen A Interferon IFN alfacon-1 InterMune
Pharmaceuticals Inc., Brisbane, CA Omega IFN Interferon IFN-.omega.
Intarcia Therapeutics IFN-.beta. and EMZ701 Interferon IFN-.beta.
and Transition EMZ701 Therapeutics Inc., Ontario, Canada REBIF
.RTM. Interferon IFN-.beta.1a Serono, Geneva, Switzerland Roferon A
Interferon IFN-.alpha.2a F. Hoffmann-La Roche LTD., Basel,
Switzerland Intron A Interferon IFN-.alpha.2b Schering-Plough
Corporation, Kenilworth, NJ Intron A and Zadaxin Interferon
IFN-.alpha.2b/.alpha.1- RegeneRx thymosin Biopharma. Inc.,
Bethesda, MD/ SciClone Pharmaceuticals Inc, San Mateo, CA Rebetron
Interferon IFN-.alpha.2b/ribavirin Schering-Plough Corporation,
Kenilworth, NJ Actimmune Interferon INF-.gamma. InterMune Inc.,
Brisbane, CA Interferon-.beta. Interferon Interferon-.beta.-1a
Serono Multiferon Interferon Long lasting IFN Viragen/Valentis
Wellferon Interferon Lymphoblastoid GlaxoSmithKline plc,
IFN-.alpha.n1 Uxbridge, UK Omniferon Interferon natural IFN-.alpha.
Viragen Inc., Plantation, FL Pegasys Interferon PEGylated IFN- F.
Hoffmann-La .alpha.2a Roche LTD, Basel, Switzerland Pegasys and
Ceplene Interferon PEGylated IFN- Maxim .alpha.2a/immune
Pharmaceuticals Inc., modulator San Diego, CA Pegasys and Ribavirin
Interferon PEGylated IFN- F. Hoffmann-La .alpha.2a/ribavirin Roche
LTD, Basel, Switzerland PEG-Intron Interferon PEGylated IFN-
Schering-Plough .alpha.2b Corporation, Kenilworth, NJ
PEG-Intron/Ribavirin Interferon PEGylated IFN- Schering-Plough
.alpha.2b/ribavirin Corporation, Kenilworth, NJ IP-501 Liver
protection Antifibrotic Indevus Pharmaceuticals Inc., Lexington, MA
IDN-6556 Liver protection Caspase inhibitor Idun Pharmaceuticals
Inc., San Diego, CA ITMN-191 (R-7227) Antiviral Serine protease
InterMune inhibitor Pharmaceuticals Inc., Brisbane, CA GL-59728
Antiviral NS5B replicase Genelabs inhibitor ANA-971 Antiviral TLR-7
agonist Anadys Boceprevir Antiviral Serine protease Schering-Plough
inhibitor TMS-435 Antiviral Serine protease Tibotec BVBA, inhibitor
Mechelen, Belgium BI-201335 Antiviral Serine protease Boehringer
Ingelheim inhibitor Pharma KG, Ingelheim, Germany MK-7009 Antiviral
Serine protease Merck inhibitor PF-00868554 Antiviral Replicase
inhibitor Pfizer ANA598 Antiviral Non-Nucleoside Anadys NS5B
polymerase Pharmaceuticals, Inc., inhibitor San Diego, CA, USA
IDX375 Antiviral Non-Nucleoside Idenix replicase inhibitor
Pharmaceuticals, Cambridge, MA, USA BILB 1941 Antiviral NS5B
polymerase Boehringer Ingelheim inhibitor Canada Ltd R&D,
Laval, QC, Canada PSI-7851 Antiviral Nucleoside Pharmasset,
polymerase Princeton, NJ, USA inhibitor VCH-759 Antiviral NS5B
polymerase Vir .degree.Chem Pharma inhibitor VCH-916 Antiviral NS5B
polymerase Vir .degree.Chem Pharma inhibitor GS-9190 Antiviral NS5B
polymerase Gilead inhibitor Peg-interferon lamda Antiviral
Interferon ZymoGenetics/ Bristol-Myers Squibb
[0138] The compounds of the present disclosure may also be used as
laboratory reagents. Compounds may be instrumental in providing
research tools for designing of viral replication assays,
validation of animal assay systems and structural biology studies
to further enhance knowledge of the HCV disease mechanisms.
Further, the compounds of the present disclosure are useful in
establishing or determining the binding site of other antiviral
compounds, for example, by competitive inhibition.
[0139] The compounds of this disclosure may also be used to treat
or prevent viral contamination of materials and therefore reduce
the risk of viral infection of laboratory or medical personnel or
patients who come in contact with such materials, e.g., blood,
tissue, surgical instruments and garments, laboratory instruments
and garments, and blood collection or transfusion apparatuses and
materials.
[0140] This disclosure is intended to encompass compounds having
Formula (I) when prepared by synthetic processes or by metabolic
processes including those occurring in the human or animal body (in
vivo) or processes occurring in vitro.
[0141] The abbreviations used in the present application, including
particularly in the illustrative examples which follow, are
well-known to those skilled in the art. Some of the abbreviations
used are as follows: Ph for phenyl; DME for 1,2-dimethoxyethane;
TFA for trifluoroacetic acid; DCM for dichloromethane; HATU for
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate; DMF for N,N-dimethylformamide; iPr.sub.2EtN,
DIEA, or DIPEA for diisopropylethylamine; h, hr, or hrs for hrs;
MeOD for CD.sub.3OD; MeOH for methanol; min or mins for minutes;
EtOH for ethanol; Et for ethyl; DMSO for dimethylsulfoxide; rt or
RT or Rt for room temperature or retention time (context will
dictate); EDCI for 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride; Boc, boc, or BOC for tert-butoxycarbonyl; EtOAc or
EtOAc for ethyl acetate; THF for tetrahydrofuran; TBDPS for
t-butyldimethylsilyl; DMAP for 4-dimethylaminopyridine; TBAF for
tetrabutylammonium fluoride; OAc for acetate; AcOH for acetic acid;
SEMC1 for 2-(trimethylsilyl)ethoxymethyl chloride; d for days; TEA
or Et.sub.3N for triethylamine; ACN or MeCN for acetonitrile; NCS
for N-chlorosuccinimide; NBS for N-bromosuccinimide; Et.sub.2O for
diethyl ether; DCE for 1,2-dichloroethane; d for days; and DEA for
diethylamine.
[0142] The present disclosure will now be described in connection
with certain embodiments which are not intended to limit its scope.
On the contrary, the present disclosure covers all alternatives,
modifications, and equivalents as can be included within the scope
of the claims. Thus, the following examples, which include specific
embodiments, will illustrate one practice of the present
disclosure, it being understood that the examples are for the
purposes of illustration of certain embodiments and are presented
to provide what is believed to be the most useful and readily
understood description of its procedures and conceptual
aspects.
[0143] Starting materials can be obtained from commercial sources
or prepared by well-established literature methods known to those
of ordinary skill in the art.
Synthesis of Common Caps
Compound Analysis Conditions
[0144] Purity assessment and low resolution mass analysis were
conducted on a Shimadzu LC system coupled with Waters
MICROMASS.RTM. ZQ MS system. It should be noted that retention
times may vary slightly between machines. Additional LC conditions
applicable to the current section, unless noted otherwise.
Cond.-MS-W1
Column=XTERRA.RTM. 3.0.times.50 mm S7
[0145] Start % B=0
Final % B=100
[0146] Gradient time=2 min Stop time=3 min Flow Rate=5 mL/min
Wavelength=220 nm
[0147] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Cond.-MS-W2
Column=XTERRA.RTM. 3.0.times.50 mm S7
Start % B=0
Final % B=100
[0148] Gradient time=3 min Stop time=4 min Flow Rate=4 mL/min
Wavelength=220 nm
[0149] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Cond.-MS-W5
Column=XTERRA.RTM. 3.0.times.50 mm S7
Start % B=0
Final % B=30
[0150] Gradient time=2 min Stop time=3 min Flow Rate=5 mL/min
Wavelength=220 nm
[0151] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Cond.-D1
Column=XTERRA.RTM. C18 3.0.times.50 mm S7
Start % B=0
Final % B=100
[0152] Gradient time=3 min Stop time=4 min Flow Rate=4 mL/min
Wavelength=220 nm
[0153] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Cond.-D2
Column .dbd.PHENOMENEX.RTM. Luna 4.6.times.50 mm S10
Start % B=0
Final % B=100
[0154] Gradient time=3 min Stop time=4 min Flow Rate=4 mL/min
Wavelength=220 nm
[0155] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Cond.-M3
Column=XTERRA.RTM. C18 3.0.times.50 mm S7
Start % B=0
Final % B=40
[0156] Gradient time=2 min Stop time=3 min Flow Rate=5 mL/min
Wavelength=220 nm
[0157] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Condition I
Column .dbd.PHENOMENEX.RTM. Luna 3.0.times.50 mm S10
Start % B=0
Final % B=100
[0158] Gradient time=2 min Stop time=3 min Flow Rate=4 mL/min
Wavelength=220 nm
[0159] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Condition II
Column .dbd.PHENOMENEX.RTM. Luna 4.6.times.50 mm S10
Start % B=0
Final % B=100
[0160] Gradient time=2 min Stop time=3 min Flow Rate=5 mL/min
Wavelength=220 nm
[0161] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Condition III
Column=XTERRA.RTM. C18 3.0.times.50 mm S7
Start % B=0
Final % B=100
[0162] Gradient time=3 min Stop time=4 min Flow Rate=4 mL/min
Wavelength=220 nm
[0163] Solvent A=0.1% TFA in 10% methanol/90% H.sub.2O Solvent
B=0.1% TFA in 90% methanol/10% H.sub.2O
Cap-1
##STR00006##
[0164] (R)-2-(Dimethylamino)-2-phenylacetic acid
[0165] A suspension of 10% Pd/C (2.0 g) in methanol (10 mL) was
added to a mixture of (R)-2-phenylglycine (10 g, 66.2 mmol),
formaldehyde (33 mL of 37% wt. in water), 1N HCl (30 mL) and
methanol (30 mL), and exposed to H.sub.2 (60 psi) for 3 hours. The
reaction mixture was filtered through diatomaceous earth
(CELITE.RTM.), and the filtrate was concentrated in vacuo. The
resulting crude material was recrystallized from isopropanol to
provide the HCl salt of Cap-1 as a white needle (4.0 g). Optical
rotation: -117.1.degree. [c=9.95 mg/mL in H.sub.2O; .lamda.=589
nm]. .sup.1H NMR (DMSO-d.sub.6, 8=2.5 ppm, 500 MHz): .delta.
7.43-7.34 (m, 5H), 4.14 (s, 1H), 2.43 (s, 6H); LC (Condition I):
RT=0.25; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.14NO.sub.2 180.10; found 180.17; HRMS: Anal. Calcd.
for [M+H].sup.+ C.sub.10H.sub.14NO.sub.2 180.1025; found
180.1017.
Cap-2
##STR00007##
[0166] (R)-2-(Diethylamino)-2-phenylacetic acid
[0167] NaBH.sub.3CN (6.22 g, 94 mmol) was added in portions over a
few minutes to a cooled (ice/water) mixture of (R)-2-Phenylglycine
(6.02 g, 39.8 mmol) and methanol (100 mL), and stirred for 5
minutes. Acetaldehyde (10 mL) was added dropwise over 10 minutes
and stirring was continued at the same cooled temperature for 45
minutes and at ambient temperature for .about.6.5 hours. The
reaction mixture was cooled back with ice-water bath, treated with
water (3 mL) and then quenched with a dropwise addition of
concentrated HCl over .about.45 minutes until the pH of the mixture
was .about.1.5-2.0. The cooling bath was removed and the stirring
was continued while adding concentrated HCl in order to maintain
the pH of the mixture around 1.5-2.0. The reaction mixture was
stirred overnight, filtered to remove the white suspension, and the
filtrate was concentrated in vacuo. The crude material was
recrystallized from ethanol to afford the HCl salt of Cap-2 as a
shining white solid in two crops (crop-1: 4.16 g; crop-2: 2.19 g).
.sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): 10.44 (1.00,
br s, 1H), 7.66 (m, 2H), 7.51 (m, 3H), 5.30 (s, 1H), 3.15 (br m,
2H), 2.98 (br m, 2H), 1.20 (app br s, 6H). Crop-1:
[.alpha.].sup.25-102.21.degree. (c=0.357, H.sub.2O); crop-2:
[.alpha.].sup.25 -99.7.degree. (c=0.357, H.sub.2O). LC (Condition
I): RT=0.43 min; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.12H.sub.18NO.sub.2: 208.13; found 208.26.
Cap-3
##STR00008##
[0169] Acetaldehyde (5.0 mL, 89.1 mmol) and a suspension of 10%
Pd/C (720 mg) in methanol/H.sub.2O (4 mL/1 mL) was sequentially
added to a cooled (.about.15.degree. C.) mixture of
(R)-2-phenylglycine (3.096 g, 20.48 mmol), 1N HCl (30 mL) and
methanol (40 mL). The cooling bath was removed and the reaction
mixture was stirred under a balloon of H.sub.2 for 17 hours. An
additional acetaldehyde (10 mL, 178.2 mmol) was added and stirring
continued under H.sub.2 atmosphere for 24 hours [Note: the supply
of H.sub.2 was replenished as needed throughout the reaction]. The
reaction mixture was filtered through diatomaceous earth
(CELITE.RTM.), and the filtrate was concentrated in vacuo. The
resulting crude material was recrystallized from isopropanol to
provide the HCl salt of (R)-2-(ethylamino)-2-phenylacetic acid as a
shining white solid (2.846 g). .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5 ppm, 400 MHz): .delta. 14.15 (br s, 1H), 9.55 (br s,
2H), 7.55-7.48 (m, 5H), 2.88 (br m, 1H), 2.73 (br m, 1H), 1.20 (app
t, J=7.2, 3H). LC (Condition I): RT=0.39 min; >95% homogeneity
index; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.14NO.sub.2: 180.10; found 180.18.
[0170] A suspension of 10% Pd/C (536 mg) in methanol/H.sub.2O (3
mL/1 mL) was added to a mixture of
(R)-2-(ethylamino)-2-phenylacetic acid/HCl (1.492 g, 6.918 mmol),
formaldehyde (20 mL of 37% wt. in water), 1N HCl (20 mL) and
methanol (23 mL). The reaction mixture was stirred under a balloon
of H.sub.2 for .about.72 hours, where the H.sub.2 supply was
replenished as needed. The reaction mixture was filtered through
diatomaceous earth (CELITE.RTM.) and the filtrate was concentrated
in vacuo. The resulting crude material was recrystallized from
isopropanol (50 mL) to provide the HCl salt of Cap-3 as a white
solid (985 mg). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 400
MHz): .delta. 10.48 (br s, 1H), 7.59-7.51 (m, 5H), 5.26 (s, 1H),
3.08 (app br s, 2H), 2.65 (br s, 3H), 1.24 (br m, 3H). LC
(Condition I): RT=0.39 min; >95% homogeneity index; LC-MS: Anal.
Calcd. for [M+H].sup.+ C.sub.11H.sub.16NO.sub.2: 194.12; found
194.18; HRMS: Anal. Calcd. for [M+H].sup.+
C.sub.11H.sub.16NO.sub.2: 194.1180; found 194.1181.
Cap-4
##STR00009##
[0171] (R)-2-(Methoxycarbonylamino)-2-phenylacetic acid
[0172] ClCO.sub.2Me (3.2 mL, 41.4 mmol) was added dropwise to a
cooled (ice/water) THF (410 mL) semi-solution of (R)-tert-butyl
2-amino-2-phenylacetate/HCl (9.877 g, 40.52 mmol) and
diisopropylethylamine (14.2 mL, 81.52 mmol) over 6 min, and stirred
at similar temperature for 5.5 hours. The volatile component was
removed in vacuo, and the residue was partitioned between water
(100 mL) and ethyl acetate (200 mL). The organic layer was washed
with 1N HCl (25 mL) and saturated NaHCO.sub.3 solution (30 mL),
dried (MgSO.sub.4), filtered, and concentrated in vacuo. The
resultant colorless oil was triturated from hexanes, filtered and
washed with hexanes (100 mL) to provide (R)-tert-butyl
2-(methoxycarbonylamino)-2-phenylacetate as a white solid (7.7 g).
.sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): 7.98 (d,
J=8.0, 1H), 7.37-7.29 (m, 5H), 5.09 (d, J=8, 1H), 3.56 (s, 3H),
1.33 (s, 9H). LC (Condition I): RT=1.53 min; .about.90% homogeneity
index; LC-MS: Anal. Calcd. for [M+Na].sup.+
C.sub.14H.sub.16NNaO.sub.4: 288.12; found 288.15.
[0173] TFA (16 mL) was added dropwise to a cooled (ice/water)
CH.sub.2Cl.sub.2 (160 mL) solution of the above product over 7
minutes, and the cooling bath was removed and the reaction mixture
was stirred for 20 hours. Since the deprotection was still not
complete, an additional TFA (1.0 mL) was added and stirring
continued for an additional 2 hours. The volatile component was
removed in vacuo, and the resulting oil residue was treated with
diethyl ether (15 mL) and hexanes (12 mL) to provide a precipitate.
The precipitate was filtered and washed with diethyl ether/hexanes
(.about.1:3 ratio; 30 mL) and dried in vacuo to provide Cap-4 as a
fluffy white solid (5.57 g). Optical rotation: -176.9.degree.
[c=3.7 mg/mL in H.sub.2O; .lamda.=589 nm]. .sup.1H NMR
(DMSO-d.sub.6, 8=2.5 ppm, 400 MHz): .delta. 12.84 (br s, 1H), 7.96
(d, J=8.3, 1H), 7.41-7.29 (m, 5H), 5.14 (d, J=8.3, 1H), 3.55 (s,
3H). LC (Condition I): RT=1.01 min; >95% homogeneity index;
LC-MS: Anal. Calcd. for [M+H].sup.+ C.sub.10H.sub.12NO.sub.4
210.08; found 210.17; HRMS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.12Na.sub.4 210.0766; found 210.0756.
Cap-5
##STR00010##
[0175] A mixture of (R)-2-phenylglycine (1.0 g, 6.62 mmol),
1,4-dibromobutane (1.57 g, 7.27 mmol) and Na.sub.2CO.sub.3 (2.10 g,
19.8 mmol) in ethanol (40 mL) was heated at 100.degree. C. for 21
hours. The reaction mixture was cooled to ambient temperature and
filtered, and the filtrate was concentrated in vacuo. The residue
was dissolved in ethanol and acidified with 1N HCl to pH 3-4, and
the volatile component was removed in vacuo. The resulting crude
material was purified by a reverse phase HPLC (water/methanol/TFA)
to provide the TFA salt of Cap-5 as a semi-viscous white foam (1.0
g). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5, 500 MHz) .delta. 10.68
(br s, 1H), 7.51 (m, 5H), 5.23 (s, 1H), 3.34 (app br s, 2H), 3.05
(app br s, 2H), 1.95 (app br s, 4H); RT=0.30 minutes (Condition I);
>98% homogeneity index; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.12H.sub.16NO.sub.2: 206.12; found 206.25.
Cap-6
##STR00011##
[0177] The TFA salt of Cap-6 was synthesized from
(R)-2-phenylglycine and 1-bromo-2-(2-bromoethoxy)ethane by using
the method of preparation of Cap-5. .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5, 500 MHz) .delta. 12.20 (br s, 1H), 7.50 (m, 5H), 4.92
(s, 1H), 3.78 (app br s, 4H), 3.08 (app br s, 2H), 2.81 (app br s,
2H); RT=0.32 minutes (Condition I); >98%; LC-MS: Anal. Calcd.
for [M+H].sup.+ C.sub.12H.sub.16NO.sub.3: 222.11; found 222.20;
HRMS: Anal. Calcd. for [M+H].sup.+ C.sub.12H.sub.16NO.sub.3:
222.1130; found 222.1121.
Cap-7
##STR00012##
[0179] A CH.sub.2Cl.sub.2 (200 mL) solution of p-toluenesulfonyl
chloride (8.65 g, 45.4 mmol) was added dropwise to a cooled
(-5.degree. C.) CH.sub.2Cl.sub.2 (200 mL) solution of (S)-benzyl
2-hydroxy-2-phenylacetate (10.0 g, 41.3 mmol), triethylamine (5.75
mL, 41.3 mmol) and 4-dimethylaminopyridine (0.504 g, 4.13 mmol),
while maintaining the temperature between -5.degree. C. and
0.degree. C. The reaction was stirred at 0.degree. C. for 9 hours,
and then stored in a freezer (-25.degree. C.) for 14 hours. It was
allowed to thaw to ambient temperature and washed with water (200
mL), 1N HCl (100 mL) and brine (100 mL), dried (MgSO.sub.4),
filtered, and concentrated in vacuo to provide benzyl
2-phenyl-2-(tosyloxy)acetate as a viscous oil which solidified upon
standing (16.5 g). The chiral integrity of the product was not
checked and that product was used for the next step without further
purification. .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5, 500 MHz)
.delta. 7.78 (d, J=8.6, 2H), 7.43-7.29 (m, 10H), 7.20 (m, 2H), 6.12
(s, 1H), 5.16 (d, J=12.5, 1H), 5.10 (d, J=12.5, 1H), 2.39 (s, 3H).
RT=3.00 (Condition III); >90% homogeneity index; LC-MS: Anal.
Calcd. for [M+H].sup.+ C.sub.22H.sub.20NaO.sub.5S: 419.09; found
419.04.
[0180] A THF (75 mL) solution of benzyl
2-phenyl-2-(tosyloxy)acetate (6.0 g, 15.1 mmol), 1-methylpiperazine
(3.36 mL, 30.3 mmol) and N,N-diisopropylethylamine (13.2 mL, 75.8
mmol) was heated at 65.degree. C. for 7 hours. The reaction was
allowed to cool to ambient temperature and the volatile component
was removed in vacuo. The residue was partitioned between
ethylacetate and water, and the organic layer was washed with water
and brine, dried (MgSO.sub.4), filtered, and concentrated in vacuo.
The resulting crude material was purified by flash chromatography
(silica gel, ethyl acetate) to provide benzyl
2-(4-methylpiperazin-1-yl)-2-phenylacetate as an orangish-brown
viscous oil (4.56 g). Chiral HPLC analysis (CHIRALCEL.RTM. OD-H)
indicated that the sample is a mixture of stereoisomers in a 38.2
to 58.7 ratio. The separation of the stereoisomers were effected as
follow: the product was dissolved in 120 mL of ethanol/heptane
(1:1) and injected (5 mL/injection) on chiral HPLC column (Chiracel
OJ, 5 cm ID.times.50 cm L, 20 .mu.m) eluting with 85:15
Heptane/ethanol at 75 mL/min, and monitored at 220 nm.
Stereoisomer-1 (1.474 g) and stereoisomer-2 (2.2149 g) were
retrieved as viscous oil. .sup.1H NMR (CDCl.sub.3, .delta.=7.26,
500 MHz) 7.44-7.40 (m, 2H), 7.33-7.24 (m, 6H), 7.21-7.16 (m, 2H),
5.13 (d, J=12.5, 1H), 5.08 (d, J=12.5, 1H), 4.02 (s, 1H), 2.65-2.38
(app br s, 8H), 2.25 (s, 3H). RT=2.10 (Condition III); >98%
homogeneity index; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.20H.sub.25N.sub.2O.sub.2: 325.19; found 325.20.
[0181] A methanol (10 mL) solution of either stereoisomer of benzyl
2-(4-methylpiperazin-1-yl)-2-phenylacetate (1.0 g, 3.1 mmol) was
added to a suspension of 10% Pd/C (120 mg) in methanol (5.0 mL).
The reaction mixture was exposed to a balloon of hydrogen, under a
careful monitoring, for <50 minutes. Immediately after the
completion of the reaction, the catalyst was filtered through
diatomaceous earth (CELITE.RTM.) and the filtrate was concentrated
in vacuo to provide Cap-7, contaminated with phenylacetic acid as a
tan foam (867.6 mg; mass is above the theoretical yield). The
product was used for the next step without further purification.
.sup.1H NMR (DMSO-d.sub.6, .delta.=2.5, 500 MHz) .delta. 7.44-7.37
(m, 2H), 7.37-7.24 (m, 3H), 3.92 (s, 1H), 2.63-2.48 (app. br s,
2H), 2.48-2.32 (m, 6H), 2.19 (s, 3H); RT=0.31 (Condition II);
>90% homogeneity index; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.13H.sub.16N.sub.2O.sub.2: 235.14; found 235.15; HRMS: Anal.
Calcd. for [M+H].sup.+ C.sub.13H.sub.16N.sub.2O.sub.2: 235.1447;
found 235.1440.
[0182] The synthesis of Cap-8 and Cap-9 was conducted according to
the synthesis of Cap-7 by using appropriate amines for the SN.sub.2
displacement step (i.e., 4-hydroxypiperidine for Cap-8 and
(S)-3-fluoropyrrolidine for Cap-9) and modified conditions for the
separation of the respective stereoisomeric intermediates, as
described below.
Cap-8
##STR00013##
[0184] The stereoisomeric separation of the intermediate benzyl
2-(4-hydroxypiperidin-1-yl)-2-phenyl acetate was effected by
employing the following conditions: the compound (500 mg) was
dissolved in ethanol/heptane (5 mL/45 mL). The resulting solution
was injected (5 mL/injection) on a chiral HPLC column (Chiracel OJ,
2 cm ID.times.25 cm L, 10 .mu.m) eluting with 80:20 heptane/ethanol
at 10 mL/min, monitored at 220 nm, to provide 186.3 mg of
stereoisomer-1 and 209.1 mg of stereoisomer-2 as light-yellow
viscous oils. These benzyl ester was hydrogenolysed according to
the preparation of Cap-7 to provide Cap-8: .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5, 500 MHz) 7.40 (d, J=7, 2H), 7.28-7.20
(m, 3H), 3.78 (s 1H), 3.46 (m, 1H), 2.93 (m, 1H), 2.62 (m, 1H),
2.20 (m, 2H), 1.70 (m, 2H), 1.42 (m, 2H). RT=0.28 (Condition II);
>98% homogeneity index; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.13H.sub.18NO.sub.3: 236.13; found 236.07; HRMS: Calcd. for
[M+H].sup.+ C.sub.13H.sub.18NO.sub.3: 236.1287; found 236.1283.
Cap-9
##STR00014##
[0186] The diastereomeric separation of the intermediate benzyl
2-((S)-3-fluoropyrrolidin-1-yl)-2-phenylacetate was effected by
employing the following conditions: the ester (220 mg) was
separated on a chiral HPLC column (Chiracel OJ-H, 0.46 cm
ID.times.25 cm L, 5 .mu.m) eluting with 95% CO.sub.2/5% methanol
with 0.1% TFA, at 10 bar pressure, 70 mL/min flow rate, and a
temperature of 35.degree. C. The HPLC elute for the respective
stereoisomers was concentrated, and the residue was dissolved in
CH.sub.2Cl.sub.2 (20 mL) and washed with an aqueous medium (10 mL
water+1 mL saturated NaHCO.sub.3 solution). The organic phase was
dried (MgSO.sub.4), filtered, and concentrated in vacuo to provide
92.5 mg of fraction-1 and 59.6 mg of fraction-2. These benzyl
esters were hydrogenolysed according to the preparation of Cap-7 to
prepare Cap-9a and Cap-9b. Cap-9a (diastereomer-1; the sample is a
TFA salt as a result of purification on a reverse phase HPLC using
H.sub.2O/methanol/TFA solvent): .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5, 400 MHz) 7.55-7.48 (m, 5H), 5.38 (d of m, J=53.7, 1H),
5.09 (br s, 1H), 3.84-2.82 (br m, 4H), 2.31-2.09 (m, 2H). RT=0.42
(Condition I); >95% homogeneity index; LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.12H.sub.15FNO.sub.2: 224.11; found 224.14; Cap-9b
(diastereomer-2): .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5, 400 MHz)
7.43-7.21 (m, 5H), 5.19 (d of m, J=55.9, 1H), 3.97 (s, 1H),
2.95-2.43 (m, 4H), 2.19-1.78 (m, 2H). RT=0.44 (Condition I); LC-MS:
Anal. Calcd. for [M+H].sup.+ C.sub.12H.sub.15FNO.sub.2: 224.11;
found 224.14.
Cap-10
##STR00015##
[0188] To a solution of D-proline (2.0 g, 17 mmol) and formaldehyde
(2.0 mL of 37% wt. in H.sub.2O) in methanol (15 mL) was added a
suspension of 10% Pd/C (500 mg) in methanol (5 mL). The mixture was
stirred under a balloon of hydrogen for 23 hours. The reaction
mixture was filtered through diatomaceous earth (CELITE.RTM.) and
concentrated in vacuo to provide Cap-10 as an off-white solid (2.15
g). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5, 500 MHz) 3.42 (m, 1H),
3.37 (dd, J=9.4, 6.1, 1H), 2.85-2.78 (m, 1H), 2.66 (s, 3H),
2.21-2.13 (m, 1H), 1.93-1.84 (m, 2H), 1.75-1.66 (m, 1H). RT=0.28
(Condition II); >98% homogeneity index; LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.6H.sub.12NO.sub.2: 130.09; found 129.96.
Cap-11
##STR00016##
[0190] A mixture of (2S,4R)-4-fluoropyrrolidine-2-carboxylic acid
(0.50 g, 3.8 mmol), formaldehyde (0.5 mL of 37% wt. in H.sub.2O),
12 N HCl (0.25 mL) and 10% Pd/C (50 mg) in methanol (20 mL) was
stirred under a balloon of hydrogen for 19 hours. The reaction
mixture was filtered through diatomaceous earth (CELITE.RTM.) and
the filtrate was concentrated in vacuo. The residue was
recrystallized from isopropanol to provide the HCl salt of Cap-11
as a white solid (337.7 mg). .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5, 500 MHz) 5.39 (d m, J=53.7, 1H), 4.30 (m, 1H), 3.90
(ddd, J=31.5, 13.5, 4.5, 1H), 3.33 (dd, J=25.6, 13.4, 1H), 2.85 (s,
3H), 2.60-2.51 (m, 1H), 2.39-2.26 (m, 1H). RT=0.28 (Condition II);
>98% homogeneity index; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.6H.sub.11FNO.sub.2: 148.08; found 148.06.
Cap-12 (Same as Cap-52)
##STR00017##
[0191] (S)-2-(Methoxycarbonylamino)propanoic acid
[0192] L-Alanine (2.0 g, 22.5 mmol) was dissolved in 10% aqueous
sodium carbonate solution (50 mL), and a THF (50 mL) solution of
methyl chloroformate (4.0 mL) was added to it. The reaction mixture
was stirred under ambient conditions for 4.5 hours and concentrated
in vacuo. The resulting white solid was dissolved in water and
acidified with 1N HCl to a pH .about.2-3. The resulting solutions
was extracted with ethyl acetate (3.times.100 mL), and the combined
organic phase was dried (Na.sub.2SO.sub.4), filtered, and
concentrated in vacuo to provide a colorless oil (2.58 g). 500 mg
of this material was purified by a reverse phase HPLC
(H.sub.2O/methanol/TFA) to provide 150 mg of Cap-12 as a colorless
oil. .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5, 500 MHz) 7.44 (d,
J=7.3, 0.8H), 7.10 (br s, 0.2H), 3.97 (m, 1H), 3.53 (s, 3H), 1.25
(d, J=7.3, 3H).
Cap-13
##STR00018##
[0194] A mixture of L-alanine (2.5 g, 28 mmol), formaldehyde (8.4
g, 37 wt. %), 1N HCl (30 mL) and 10% Pd/C (500 mg) in methanol (30
mL) was stirred under a hydrogen atmosphere (50 psi) for 5 hours.
The reaction mixture was filtered through diatomaceous earth
(CELITE.RTM.) and the filtrate was concentrated in vacuo to provide
the HCl salt of Cap-13 as an oil which solidified upon standing
under vacuum (4.4 g; the mass is above theoretical yield). The
product was used without further purification. .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5, 500 MHz) .delta. 12.1 (br s, 1H), 4.06
(q, J=7.4, 1H), 2.76 (s, 6H), 1.46 (d, J=7.3, 3H).
Cap-14
##STR00019##
[0195] (R)-2-Phenyl-2-(piperidin-1-yl)acetic acid
[0196] Step 1: A mixture of (R)-(-)-D-phenylglycine tert-butyl
ester (3.00 g, 12.3 mmol), NaBH.sub.3CN (0.773 g, 12.3 mmol), KOH
(0.690 g, 12.3 mmol) and acetic acid (0.352 mL, 6.15 mmol) were
stirred in methanol at 0.degree. C. To this mixture was added
glutaric dialdehyde (2.23 mL, 12.3 mmol) dropwise over 5 minutes.
The reaction mixture was stirred as it was allowed to warm to
ambient temperature and stirring was continued at the same
temperature for 16 hours. The solvent was subsequently removed and
the residue was partitioned with 10% aqueous NaOH and ethyl
acetate. The organic phase was separated, dried (MgSO.sub.4),
filtered and concentrated to dryness to provide a clear oil. This
material was purified by reverse-phase preparative HPLC
(Primesphere C-18, 30.times.100 mm; CH.sub.3CN--H.sub.2O-0.1% TFA)
to give the intermediate ester (2.70 g, 56%) as a clear oil.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.53-7.44 (m, 3H),
7.40-7.37 (m, 2H), 3.87 (d, J=10.9 Hz, 1H), 3.59 (d, J=10.9 Hz,
1H), 2.99 (t, J=11.2 Hz, 1H), 2.59 (t, J=11.4 Hz, 1H), 2.07-2.02
(m, 2H), 1.82 (d, J=1.82 Hz, 3H), 1.40 (s, 9H). LC-MS: Anal. Calcd.
for C.sub.17H.sub.25NO.sub.2: 275; found: 276 (M+H).sup.+.
[0197] Step 2: To a stirred solution of the intermediate ester
(1.12 g, 2.88 mmol) in dichloromethane (10 mL) was added TFA (3
mL). The reaction mixture was stirred at ambient temperature for 4
hours and then it was concentrated to dryness to give a light
yellow oil. The oil was purified using reverse-phase preparative
HPLC (Primesphere C-18, 30.times.100 mm; CH.sub.3CN--H.sub.2O-0.1%
TFA). The appropriate fractions were combined and concentrated to
dryness in vacuo. The residue was then dissolved in a minimum
amount of methanol and applied to applied to MCX LP extraction
cartridges (2.times.6 g). The cartridges were rinsed with methanol
(40 mL) and then the desired compound was eluted using 2M ammonia
in methanol (50 mL). Product-containing fractions were combined and
concentrated and the residue was taken up in water. Lyophilization
of this solution provided the title compound (0.492 g, 78%) as a
light yellow solid. .sup.1H NMR (DMSO-d.sub.6) .delta. 7.50 (s,
5H), 5.13 (s, 1H), 3.09 (br s, 2H), 2.92-2.89 (m, 2H), 1.74 (m,
4H), 1.48 (br s, 2H). LC-MS: Anal. Calcd. for
C.sub.13H.sub.17NO.sub.2: 219; found: 220 (M+H).sup.+.
Cap-15
##STR00020##
[0199] Step 1: (S)-1-Phenylethyl 2-bromo-2-phenylacetate. To a
mixture of .alpha.-bromophenylacetic acid (10.75 g, 0.050 mol),
(S)-(-)-1-phenylethanol (7.94 g, 0.065 mol) and DMAP (0.61 g, 5.0
mmol) in dry dichloromethane (100 mL) was added solid EDCI (12.46
g, 0.065 mol) all at once. The resulting solution was stirred at
room temperature under Ar for 18 hours and then it was diluted with
ethyl acetate, washed (H.sub.2O.times.2, brine), dried
(Na.sub.2SO.sub.4), filtered, and concentrated to give a pale
yellow oil. Flash chromatography (SiO.sub.2/hexane-ethyl acetate,
4:1) of this oil provided the title compound (11.64 g, 73%) as a
white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.53-7.17
(m, 10H), 5.95 (q, J=6.6 Hz, 0.5H), 5.94 (q, J=6.6 Hz, 0.5H), 5.41
(s, 0.5H), 5.39 (s, 0.5H), 1.58 (d, J=6.6 Hz, 1.5H), 1.51 (d, J=6.6
Hz, 1.5H).
[0200] Step 2: (S)-1-Phenylethyl
(R)-2-(4-hydroxy-4-methylpiperidin-1-yl)-2-phenylacetate. To a
solution of (S)-1-phenylethyl 2-bromo-2-phenylacetate (0.464 g,
1.45 mmol) in THF (8 mL) was added triethylamine (0.61 mL, 4.35
mmol), followed by tetrabutylammonium iodide (0.215 g, 0.58 mmol).
The reaction mixture was stirred at room temperature for 5 minutes
and then a solution of 4-methyl-4-hydroxypiperidine (0.251 g, 2.18
mmol) in THF (2 mL) was added. The mixture was stirred for 1 hour
at room temperature and then it was heated at 55-60.degree. C. (oil
bath temperature) for 4 hours.
[0201] The cooled reaction mixture was then diluted with ethyl
acetate (30 mL), washed (H.sub.2O x2, brine), dried (MgSO.sub.4),
filtered and concentrated. The residue was purified by silica gel
chromatography (0-60% ethyl acetate-hexane) to provide first the
(S,R)-isomer of the title compound (0.306 g, 60%) as a white solid
and then the corresponding (S,S)-isomer (0.120 g, 23%), also as a
white solid. (S,R)-isomer: .sup.1H NMR (CD.sub.3OD) .delta.
7.51-7.45 (m, 2H), 7.41-7.25 (m, 8H), 5.85 (q, J=6.6 Hz, 1H), 4.05
(s, 1H), 2.56-2.45 (m, 2H), 2.41-2.29 (m, 2H), 1.71-1.49 (m, 4H),
1.38 (d, J=6.6 Hz, 3H), 1.18 (s, 3H). LC-MS: Anal. Calcd. for
C.sub.22H.sub.27NO.sub.3: 353; found: 354 (M+H).sup.+.
(S,S)-isomer: .sup.1H NMR (CD.sub.3OD) .delta. 7.41-7.30 (m, 5H),
7.20-7.14 (m, 3H), 7.06-7.00 (m, 2H), 5.85 (q, J=6.6 Hz, 1H), 4.06
(s, 1H), 2.70-2.60 (m, 1H), 2.51 (dt, J=6.6, 3.3 Hz, 1H), 2.44-2.31
(m, 2H), 1.75-1.65 (m, 1H), 1.65-1.54 (m, 3H), 1.50 (d, J=6.8 Hz,
3H), 1.20 (s, 3H). LC-MS: Anal. Calcd. for
C.sub.22H.sub.27NO.sub.3: 353; found: 354 (M+H).sup.+.
[0202] Step 3:
(R)-2-(4-Hydroxy-4-methylpiperidin-1-yl)-2-phenylacetic acid. To a
solution of (S)-1-phenylethyl
(R)-2-(4-hydroxy-4-methylpiperidin-1-yl)-2-phenylacetate (0.185 g,
0.52 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid
(1 mL) and the mixture was stirred at room temperature for 2 hours.
The volatiles were subsequently removed in vacuo and the residue
was purified by reverse-phase preparative HPLC (Primesphere C-18,
20.times.100 mm; CH.sub.3CN--H.sub.2O-0.1% TFA) to give the title
compound (as TFA salt) as a pale bluish solid (0.128 g, 98%).
LC-MS: Anal. Calcd. for C.sub.14H.sub.19NO.sub.3: 249; found: 250
(M+H).sup.+.
Cap-16
##STR00021##
[0204] Step 1: (S)-1-Phenylethyl 2-(2-fluorophenyl)acetate. A
mixture of 2-fluorophenylacetic acid (5.45 g, 35.4 mmol),
(S)-1-phenylethanol (5.62 g, 46.0 mmol), EDCI (8.82 g, 46.0 mmol)
and DMAP (0.561 g, 4.60 mmol) in CH.sub.2Cl.sub.2 (100 mL) was
stirred at room temperature for 12 hours. The solvent was then
concentrated and the residue partitioned with H.sub.2O-ethyl
acetate. The phases were separated and the aqueous layer
back-extracted with ethyl acetate (2.times.). The combined organic
phases were washed (H.sub.2O, brine), dried (Na.sub.2SO.sub.4),
filtered, and concentrated in vacuo. The residue was purified by
silica gel chromatography (BIOTAGE.RTM./0-20% ethyl acetate-hexane)
to provide the title compound as a colorless oil (8.38 g, 92%).
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.32-7.23 (m, 7H),
7.10-7.04 (m, 2), 5.85 (q, J=6.5 Hz, 1H), 3.71 (s, 2H), 1.48 (d,
J=6.5 Hz, 3H).
[0205] Step 2: (R)--((S)-1-Phenylethyl)
2-(2-fluorophenyl)-2-(piperidin-1-yl)acetate.
[0206] To a solution of (S)-1-phenylethyl 2-(2-fluorophenyl)acetate
(5.00 g, 19.4 mmol) in THF (1200 mL) at 0.degree. C. was added DBU
(6.19 g, 40.7 mmol) and the solution was allowed to warm to room
temperature while stirring for 30 minutes. The solution was then
cooled to -78.degree. C. and a solution of CBr.sub.4 (13.5 g, 40.7
mmol) in THF (100 mL) was added and the mixture was allowed to warm
to -10.degree. C. and stirred at this temperature for 2 hours. The
reaction mixture was quenched with saturated aq. NH.sub.4Cl and the
layers were separated. The aqueous layer was back-extracted with
ethyl acetate (2.times.) and the combined organic phases were
washed (H.sub.2O, brine), dried (Na.sub.2SO.sub.4), filtered, and
concentrated in vacuo. To the residue was added piperidine (5.73
mL, 58.1 mmol) and the solution was stirred at room temperature for
24 hours. The volatiles were then concentrated in vacuo and the
residue was purified by silica gel chromatography
(BIOTAGE.RTM./0-30% diethyl ether-hexane) to provide a pure mixture
of diastereomers (2:1 ratio by .sup.1H NMR) as a yellow oil (2.07
g, 31%), along with unreacted starting material (2.53 g, 51%).
Further chromatography of the diastereomeric mixture
(BIOTAGE.RTM./0-10% diethyl ether-toluene) provided the title
compound as a colorless oil (0.737 g, 11%). .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.52 (ddd, J=9.4, 7.6, 1.8 Hz, 1H), 7.33-7.40
(m, 1), 7.23-7.23 (m, 4H), 7.02-7.23 (m, 4H), 5.86 (q, J=6.6 Hz,
1H), 4.45 (s, 1H), 2.39-2.45 (m, 4H), 1.52-1.58 (m, 4H), 1.40-1.42
(m, 1H), 1.38 (d, J=6.6 Hz, 3H). LC-MS: Anal. Calcd. for
C.sub.21H.sub.24FNO.sub.2: 341; found: 342 (M+H).sup.+.
[0207] Step 3: (R)-2-(2-Fluorophenyl)-2-(piperidin-1-yl)acetic
acid. A mixture of
(R)--((S)-1-phenylethyl)2-(2-fluorophenyl)-2-(piperidin-1-yl)acetate
(0.737 g, 2.16 mmol) and 20% Pd(OH).sub.2/C (0.070 g) in ethanol
(30 mL) was hydrogenated at room temperature and atmospheric
pressure (H.sub.2 balloon) for 2 hours. The solution was then
purged with Ar, filtered through diatomaceous earth (CELITE.RTM.),
and concentrated in vacuo. This provided the title compound as a
colorless solid (0.503 g, 98%). .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.65 (ddd, J=9.1, 7.6, 1.5 Hz, 1H), 7.47-7.53 (m, 1H),
7.21-7.30 (m, 2H), 3.07-3.13 (m, 4H), 1.84 (br s, 4H), 1.62 (br s,
2H). LC-MS: Anal. Calcd. for C.sub.13H.sub.16FNO.sub.2: 237; found:
238 (M+H).sup.+.
Cap-17
##STR00022##
[0209] Step 1: (S)-1-Phenylethyl
(R)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-2-phenylacetate. To a
solution of (S)-1-phenylethyl 2-bromo-2-phenylacetate (1.50 g, 4.70
mmol) in THF (25 mL) was added triethylamine (1.31 mL, 9.42 mmol),
followed by tetrabutylammonium iodide (0.347 g, 0.94 mmol). The
reaction mixture was stirred at room temperature for 5 minutes and
then a solution of 4-phenyl-4-hydroxypiperidine (1.00 g, 5.64 mmol)
in THF (5 mL) was added. The mixture was stirred for 16 hours and
then it was diluted with ethyl acetate (100 mL), washed (H.sub.2O
x2, brine), dried (MgSO.sub.4), filtered and concentrated. The
residue was purified on a silica gel column (0-60% ethyl
acetate-hexane) to provide an approximately 2:1 mixture of
diastereomers, as judged by .sup.1H NMR. Separation of these
isomers was performed using supercritical fluid chromatography
(CHIRALCEL.RTM. OJ-H, 30.times.250 mm; 20% ethanol in CO.sub.2 at
35.degree. C.), to give first the (R)-isomer of the title compound
(0.534 g, 27%) as a yellow oil and then the corresponding
(S)-isomer (0.271 g, 14%), also as a yellow oil. (S,R)-isomer:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.55-7.47 (m, 4H),
7.44-7.25 (m, 10H), 7.25-7.17 (m, 1H), 5.88 (q, J=6.6 Hz, 1H), 4.12
(s, 1H), 2.82-2.72 (m, 1H), 2.64 (dt, J=11.1, 2.5 Hz, 1H),
2.58-2.52 (m, 1H), 2.40 (dt, J=11.1, 2.5 Hz, 1H), 2.20 (dt, J=12.1,
4.6 Hz, 1H), 2.10 (dt, J=12.1, 4.6 Hz, 1H), 1.72-1.57 (m, 2H), 1.53
(d, J=6.5 Hz, 3H). LC-MS: Anal. Calcd. for
C.sub.27H.sub.29NO.sub.3: 415; found: 416 (M+H).sup.+;
(S,S)-isomer: H.sup.1NMR (400 MHz, CD.sub.3OD) .delta. 7.55-7.48
(m, 2H), 7.45-7.39 (m, 2H), 7.38-7.30 (m, 5H), 7.25-7.13 (m, 4H),
7.08-7.00 (m, 2H), 5.88 (q, J=6.6 Hz, 1H), 4.12 (s, 1H), 2.95-2.85
(m, 1H), 2.68 (dt, J=11.1, 2.5 Hz, 1H), 2.57-2.52 (m, 1H), 2.42
(dt, J=11.1, 2.5 Hz, 1H), 2.25 (dt, J=12.1, 4.6 Hz, 1H), 2.12 (dt,
J=12.1, 4.6 Hz, 1H), 1.73 (dd, J=13.6, 3.0 Hz, 1H), 1.64 (dd,
J=13.6, 3.0 Hz, 1H), 1.40 (d, J=6.6 Hz, 3H). LC-MS: Anal. Calcd.
for C.sub.27H.sub.29NO.sub.3: 415; found: 416 (M+H).sup.+.
[0210] The following esters were prepared in similar fashion:
TABLE-US-00002 Intermediate-17a ##STR00023## Diastereomer 1:
.sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 1.36 (d, J = 6.41
Hz, 3H) 2.23-2.51 (m, 4H) 3.35 (s, 4H) 4.25 (s, 1H) 5.05 (s, 2H)
5.82 (d, J = 6.71 Hz, 1H) 7.15-7.52 (m, 15H). LC-MS: Anal. Calcd.
for: C.sub.28H.sub.30N.sub.2O.sub.4 458.22; found: 459.44 (M +
H).sup.+. Diastereomer 2: .sup.1H NMR (500 MHz, DMSO- d.sub.6)
.delta. ppm 1.45 (d, J = 6.71 Hz, 3H) 2.27-2.44 (m, 4H) 3.39 (s,
4H) 4.23 (s, 1H) 5.06 (s, 2H) 5.83 (d, J = 6.71 Hz, 1H) 7.12 (dd, J
= 6.41, 3.05 Hz, 2H) 7.19-7.27 (m, 3H) 7.27-7.44 (m, 10H). LC-MS:
Anal. Calcd. for: C.sub.28H.sub.30N.sub.2O.sub.4 458.22; found:
459.44 (M + H).sup.+. Intermediate-17b ##STR00024## Diastereomer 1:
RT = 11.76 minutes (Condition II); LC-MS: Anal. Calcd. for:
C.sub.20H.sub.22N.sub.2O.sub.3 338.16; found: 339.39 (M + H).sup.+.
Diastereomer 2: RT = 10.05 minutes (Condition II). LC-MS: Anal.
Calcd. for: C.sub.20H.sub.22N.sub.2O.sub.3 338.16; found: 339.39 (M
+ H).sup.+. Intermediate-17c ##STR00025## Diastereomer 1: T.sub.R =
4.55 minutes (Condition I); LC-MS: Anal. Calcd. for:
C.sub.21H.sub.26N.sub.2O.sub.2 338.20; found: 339.45 (M + H).sup.+.
Diastereomer 2: T.sub.R = 6.00 minutes (Condition I). LC-MS: Anal.
Calcd. for: C.sub.21H.sub.26N.sub.2O.sub.2 338.20; found: 339.45 (M
+ H).sup.+. Intermediate-17d ##STR00026## Diastereomer 1: RT = 7.19
minutes (Condition I); LC-MS: Anal. Calcd. for:
C.sub.27H.sub.29NO.sub.2 399.22; found: 400.48 (M + H).sup.+.
Diastereomer 2: RT = 9.76 minutes (Condition I); LC-MS: Anal.
Calcd. for: C.sub.27H.sub.29NO.sub.2 399.22; found: 400.48 (M +
H).sup.+.
Chiral SFC Conditions for Determining Retention Time:
Condition I
[0211] Column. CHIRALPAK.RTM. AD-H Column, 4.62.times.50 mm, 5 nm
Solvents: 90% CO.sub.2-10% methanol with 0.1% DEA
Temp: 35.degree. C.
Pressure: 150 bar
[0212] Flow rate: 2.0 mL/min. UV monitored at 220 nm
[0213] Injection: 1.0 mg/3 mL methanol
Condition II
[0214] Column. CHIRALCEL.RTM. OD-H Column, 4.62.times.50 mm, 5 nm
Solvents: 90% CO.sub.2-10% methanol with 0.1% DEA
Temp: 35.degree. C.
Pressure: 150 bar
[0215] Flow rate: 2.0 mL/min. UV monitored at 220 nm Injection: 1.0
mg/mL methanol
[0216] Cap-17, Step 2:
(R)-2-(4-Hydroxy-4-phenylpiperidin-1-yl)-2-phenylacetic acid. To a
solution of (S)-1-phenylethyl
(R)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-2-phenylacetate (0.350 g,
0.84 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid
(1 mL) and the mixture was stirred at room temperature for 2 hours.
The volatiles were subsequently removed in vacuo and the residue
was purified by reverse-phase preparative HPLC (Primesphere C-18,
20.times.100 mm; CH.sub.3CN--H.sub.2O-0.1% TFA) to give the title
compound (as TFA salt) as a white solid (0.230 g, 88%). LC-MS:
Anal. Calcd. for C.sub.19H.sub.21NO.sub.3: 311.15; found: 312
(M+H).sup.+.
[0217] The following carboxylic acids were prepared in optically
pure form in a similar fashion:
TABLE-US-00003 Cap-17a ##STR00027## RT = 2.21 (Condition II);
.sup.1H NMR (500 MHz, DMSO- d.sub.6) .delta. ppm 2.20-2.35 (m, 2H)
2.34-2.47 (m, 2H) 3.37 (s, 4H) 3.71 (s, 1H) 5.06 (s, 2H) 7.06-7.53
(m, 10H). LC- MS: Anal. Calcd. for: C.sub.20H.sub.22N.sub.2O.sub.4
354.16; found: 355.38 (M + H).sup.+. Cap-17b ##STR00028## RT = 0.27
(Condition III); LC-MS: Anal. Calcd. for:
C.sub.12H.sub.14N.sub.2O.sub.3 234.10; found: 235.22 (M + H).sup.+.
Cap-17c ##STR00029## RT = 0.48 (Condition II); LC-MS: Anal. Calcd.
for: C.sub.13H.sub.18N.sub.2O.sub.2 234.14; found: 235.31 (M +
H).sup.+. Cap-17d ##STR00030## RT = 2.21 (Condition I); LC-MS:
Anal. Calcd. for: C.sub.19H.sub.21NO.sub.2 295.16; found: 296.33 (M
+ H).sup.+.
LC-MS Conditions for Determining Retention Time:
Condition I
[0218] Column. PHENOMENEX.RTM. Luna 4.6.times.50 mm S10
Start % B=0
Final % B=100
Gradient Time=4 min
[0219] Flow Rate=4 mL/min
Wavelength=220
[0220] Solvent A=10% methanol-90% H.sub.2O-0.1% TFA Solvent B=90%
methanol-10% H.sub.2O-0.1% TFA
Condition II
[0221] Column. Waters-Sunfire 4.6.times.50 mm S5
Start % B=0
Final % B=100
Gradient Time=2 min
[0222] Flow Rate=4 mL/min
Wavelength=220
[0223] Solvent A=10% methanol-90% H.sub.2O-0.1% TFA Solvent B=90%
methanol-10% H.sub.2O-0.1% TFA
Condition III
[0224] Column. PHENOMENEX.RTM. 10.mu. 3.0.times.50 mm
Start % B=0
Final % B=100
Gradient Time=2 min
[0225] Flow Rate=4 mL/min
Wavelength=220
[0226] Solvent A=10% methanol-90% H.sub.2O-0.1% TFA Solvent B=90%
methanol-10% H.sub.2O-0.1% TFA
Cap-18
##STR00031##
[0228] Step 1: (R,S)-Ethyl 2-(4-pyridyl)-2-bromoacetate. To a
solution of ethyl 4-pyridylacetate (1.00 g, 6.05 mmol) in dry THF
(150 mL) at 0.degree. C. under argon was added DBU (0.99 mL, 6.66
mmol). The reaction mixture was allowed to warm to room temperature
over 30 minutes and then it was cooled to -78.degree. C. To this
mixture was added CBr.sub.4 (2.21 g, 6.66 mmol) and stirring was
continued at -78.degree. C. for 2 hours. The reaction mixture was
then quenched with sat. aq. NH.sub.4Cl and the phases were
separated. The organic phase was washed (brine), dried
(Na.sub.2SO.sub.4), filtered, and concentrated in vacuo. The
resulting yellow oil was immediately purified by flash
chromatography (SiO.sub.2/hexane-ethyl acetate, 1:1) to provide the
title compound (1.40 g, 95%) as a somewhat unstable yellow oil.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.62 (dd, J=4.6, 1.8 Hz,
2H), 7.45 (dd, J=4.6, 1.8 Hz, 2H), 5.24 (s, 1H), 4.21-4.29 (m, 2H),
1.28 (t, J=7.1 Hz, 3H). LC-MS: Anal. Calcd. for
C.sub.9H.sub.10BrNO.sub.2: 242, 244; found: 243, 245
(M+H).sup.+.
[0229] Step 2: (R,S)-Ethyl
2-(4-pyridyl)-2-(N,N-dimethylamino)acetate. To a solution of
(R,S)-ethyl 2-(4-pyridyl)-2-bromoacetate (1.40 g, 8.48 mmol) in DMF
(10 mL) at room temperature was added dimethylamine (2M in THF, 8.5
mL, 17.0 mmol). After completion of the reaction (as judged by thin
layer chromatography) the volatiles were removed in vacuo and the
residue was purified by flash chromatography (BIOTAGE.RTM., 40+M
SiO.sub.2 column; 50%-100% ethyl acetate-hexane) to provide the
title compound (0.539 g, 31%) as a light yellow oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.58 (d, J=6.0 Hz, 2H), 7.36 (d,
J=6.0 Hz, 2H), 4.17 (m, 2H), 3.92 (s, 1H), 2.27 (s, 6H), 1.22 (t,
J=7.0 Hz). LC-MS: Anal. Calcd. for C.sub.11H.sub.16N.sub.2O.sub.2:
208; found: 209 (M+H).sup.+.
[0230] Step 3: (R,S)-2-(4-Pyridyl)-2-(N,N-dimethylamino)acetic
acid. To a solution of (R,S)-ethyl
2-(4-pyridyl)-2-(N,N-dimethylamino)acetate (0.200 g, 0.960 mmol) in
a mixture of THF-methanol-H.sub.2O (1:1:1, 6 mL) was added powdered
LiOH (0.120 g, 4.99 mmol) at room temperature. The solution was
stirred for 3 hours and then it was acidified to pH 6 using 1N HCl.
The aqueous phase was washed with ethyl acetate and then it was
lyophilized to give the dihydrochloride of the title compound as a
yellow solid (containing LiCl). The product was used as such in
subsequent steps. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.49
(d, J=5.7 Hz, 2H), 7.34 (d, J=5.7 Hz, 2H), 3.56 (s, 1H), 2.21 (s,
6H).
[0231] The following examples were prepared in similar fashion
using the method described above:
TABLE-US-00004 Cap-19 ##STR00032## LC-MS: Anal. Calcd. for
C.sub.9H.sub.12N.sub.2O.sub.2: 180; found: 181 (M + H).sup.+.
Cap-20 ##STR00033## LC-MS: no ionization. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 8.55 (d, J = 4.3 Hz, 1H), 7.84 (app t, J = 5.3
Hz, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.37 (app t, J = 5.3 Hz, 1H),
4.35 (s, 1H), 2.60 (s, 6H). Cap-21 ##STR00034## LC-MS: Anal. Calcd.
for C.sub.9H.sub.11ClN.sub.2O.sub.2: 214, 216; found: 215, 217 (M +
H).sup.+. Cap-22 ##STR00035## LC-MS: Anal. Calcd. for
C.sub.10H.sub.12N.sub.2O.sub.4: 224; found: 225 (M + H).sup.+.
Cap-23 ##STR00036## LC-MS: Anal. Calcd. for
C.sub.14H.sub.15NO.sub.2: 229; found: 230 (M + H).sup.+. Cap-24
##STR00037## LC-MS: Anal. Calcd. for
C.sub.11H.sub.12F.sub.3NO.sub.2: 247; found: 248 (M + H).sup.+ .
Cap-25 ##STR00038## LC-MS: Anal. Calcd. for
C.sub.11H.sub.12F.sub.3NO.sub.2: 247; found: 248 (M + H).sup.+.
Cap-26 ##STR00039## LC-MS: Anal. Calcd. for
C.sub.10H.sub.12FNO.sub.2: 197; found: 198 (M + H).sup.+. Cap-27
##STR00040## LC-MS: Anal. Calcd. for C.sub.10H.sub.12FNO.sub.2:
247; found: 248 (M + H).sup.+. Cap-28 ##STR00041## LC-MS: Anal.
Calcd. for C.sub.10H.sub.12ClNO.sub.2: 213; found: 214 (M +
H).sup.+. Cap-29 ##STR00042## LC-MS: Anal. Calcd. for
C.sub.10H.sub.12ClNO.sub.2: 213; found: 214 (M + H).sup.+. Cap-30
##STR00043## LC-MS: Anal. Calcd. for C.sub.10H.sub.12ClNO.sub.2:
213; found: 214 (M + H).sup.+. Cap-31 ##STR00044## LC-MS: Anal.
Calcd. for C.sub.8H.sub.12N.sub.2O.sub.2S: 200; found: 201 (M +
H).sup.+. Cap-32 ##STR00045## LC-MS: Anal. Calcd. for
C.sub.8H.sub.11NO.sub.2S: 185; found: 186 (M + H).sup.+. Cap-33
##STR00046## LC-MS: Anal. Calcd. for C.sub.8H.sub.11NO.sub.2S: 185;
found: 186 (M + H).sup.+. Cap-34 ##STR00047## LC-MS: Anal. Calcd.
for C.sub.11H.sub.12N.sub.2O.sub.3: 220; found: 221 (M + H).sup.+.
Cap-35 ##STR00048## LC-MS: Anal. Calcd. for
C.sub.12H.sub.13NO.sub.2S: 235; found: 236 (M + H).sup.+. Cap-36
##STR00049## LC-MS: Anal. Calcd. for
C.sub.12H.sub.14N.sub.2O.sub.2S: 250; found: 251 (M + H).sup.+.
Cap-37
##STR00050##
[0233] Step 1: (R,S)-Ethyl
2-(quinolin-3-yl)-2-(N,N-dimethylamino)-acetate. A mixture of ethyl
N,N-dimethylaminoacetate (0.462 g, 3.54 mmol), K.sub.3PO.sub.4
(1.90 g, 8.95 mmol), Pd(t-Bu.sub.3P).sub.2 (0.090 g, 0.176 mmol),
3-bromoquinoline and toluene (10 mL) was degassed with a stream of
Ar bubbles for 15 minutes. The reaction mixture was then heated at
100.degree. C. for 12 hours, after which it was cooled to room
temperature and poured into H.sub.2O. The mixture was extracted
with ethyl acetate (2.times.) and the combined organic phases were
washed (H.sub.2O, brine), dried (Na.sub.2SO.sub.4), filtered, and
concentrated in vacuo.
[0234] The residue was purified first by reverse-phase preparative
HPLC (Primesphere C-18, 30.times.100 mm; CH.sub.3CN--H.sub.2O-5 mM
NH.sub.4OAc) and then by flash chromatography
(SiO.sub.2/hexane-ethyl acetate, 1:1) to provide the title compound
(0.128 g, 17%) as an orange oil. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.90 (d, J=2.0 Hz, 1H), 8.32 (d, J=2.0 Hz, 1H), 8.03-8.01
(m, 2H), 7.77 (ddd, J=8.3, 6.8, 1.5 Hz, 1H), 7.62 (ddd, J=8.3, 6.8,
1.5 Hz, 1H), 4.35 (s, 1H), 4.13 (m, 2H), 2.22 (s, 6H), 1.15 (t,
J=7.0 Hz, 3H). LC-MS: Anal. Calcd. for
C.sub.15H.sub.18N.sub.2O.sub.2: 258; found: 259 (M+H).sup.+.
[0235] Step 2: (R,S) 2-(Quinolin-3-yl)-2-(N,N-dimethylamino)acetic
acid. A mixture of (R,S)-ethyl
2-(quinolin-3-yl)-2-(N,N-dimethylamino)acetate (0.122 g, 0.472
mmol) and 6M HCl (3 mL) was heated at 100.degree. C. for 12 hours.
The solvent was removed in vacuo to provide the dihydrochloride of
the title compound (0.169 g, >100%) as a light yellow foam. The
unpurified material was used in subsequent steps without further
purification. LC-MS: Anal. Calcd. for
C.sub.13H.sub.14N.sub.2O.sub.2: 230; found: 231 (M+H).sup.+.
Cap-38
##STR00051##
[0237] Step 1:
(R)--((S)-1-Phenylethyl)2-(dimethylamino)-2-(2-fluorophenyl)acetate
and
(S)--((S)-1-Phenylethyl)2-(dimethylamino)-2-(2-fluorophenyl)acetate.
To a mixture of (RS)-2-(dimethylamino)-2-(2-fluorophenyl)acetic
acid (2.60 g, 13.19 mmol), DMAP (0.209 g, 1.71 mmol) and
(S)-1-phenylethanol (2.09 g, 17.15 mmol) in CH.sub.2Cl.sub.2 (40
mL) was added EDCI (3.29 g, 17.15 mmol) and the mixture was allowed
to stir at room temperature for 12 hours. The solvent was then
removed in vacuo and the residue partitioned with ethyl
acetate-H.sub.2O. The layers were separated, the aqueous layer was
back-extracted with ethyl acetate (2.times.) and the combined
organic phases were washed (H.sub.2O, brine), dried
(Na.sub.2SO.sub.4), filtered, and concentrated in vacuo. The
residue was purified by silica gel chromatography
(BIOTAGE.RTM./0-50% diethyl ether-hexane). The resulting pure
diastereomeric mixture was then separated by reverse-phase
preparative HPLC (Primesphere C-18, 30.times.100 mm;
CH.sub.3CN--H.sub.2O-0.1% TFA) to give first (S)-1-phenethyl
(R)-2-(dimethylamino)-2-(2-fluorophenyl)acetate (0.501 g, 13%) and
then (S)-1-phenethyl
(S)-2-(dimethylamino)-2-(2-fluorophenyl)-acetate (0.727 g. 18%),
both as their TFA salts. (S,R)-isomer: .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.65-7.70 (m, 1H), 7.55-7.60 (ddd, J=9.4, 8.1,
1.5 Hz, 1H), 7.36-7.41 (m, 2H), 7.28-7.34 (m, 5H), 6.04 (q, J=6.5
Hz, 1H), 5.60 (s, 1H), 2.84 (s, 6H), 1.43 (d, J=6.5 Hz, 3H). LC-MS:
Anal. Calcd. for C.sub.18H.sub.20FNO.sub.2: 301; found: 302
(M+H).sup.+; (S,S)-isomer: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.58-7.63 (m, 1H), 7.18-7.31 (m, 6H), 7.00 (dd, J=8.5, 1.5
Hz, 2H), 6.02 (q, J=6.5 Hz, 1H), 5.60 (s, 1H), 2.88 (s, 6H), 1.54
(d, J=6.5 Hz, 3H). LC-MS: Anal. Calcd. for
C.sub.18H.sub.20FNO.sub.2: 301; found: 302 (M+H).sup.+.
[0238] Step 2: (R)-2-(Dimethylamino)-2-(2-fluorophenyl)acetic acid.
A mixture of
(R)--((S)-1-phenylethyl)2-(dimethylamino)-2-(2-fluorophenyl)acetate
TFA salt (1.25 g, 3.01 mmol) and 20% Pd(OH).sub.2/C (0.125 g) in
ethanol (30 mL) was hydrogenated at room temperature and
atmospheric pressure (H.sub.2 balloon) for 4 hours. The solution
was then purged with Ar, filtered through diatomaceous earth
(CELITE.RTM.), and concentrated in vacuo. This gave the title
compound as a colorless solid (0.503 g, 98%). .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.53-7.63 (m, 2H), 7.33-7.38 (m, 2H), 5.36 (s,
1H), 2.86 (s, 6H). LC-MS: Anal. Calcd. for
C.sub.10H.sub.12FNO.sub.2: 197; found: 198 (M+H).sup.+.
[0239] The S-isomer could be obtained from (S)--((S)-1-phenylethyl)
2-(dimethylamino)-2-(2-fluorophenyl)acetate TFA salt in similar
fashion.
Cap-39
##STR00052##
[0241] A mixture of (R)-(2-chlorophenyl)glycine (0.300 g, 1.62
mmol), formaldehyde (35% aqueous solution, 0.80 mL, 3.23 mmol) and
20% Pd(OH).sub.2/C (0.050 g) was hydrogenated at room temperature
and atmospheric pressure (H.sub.2 balloon) for 4 hours. The
solution was then purged with Ar, filtered through diatomaceous
earth (CELITE.RTM.) and concentrated in vacuo. The residue was
purified by reverse-phase preparative HPLC (Primesphere C-18,
30.times.100 mm; CH.sub.3CN--H.sub.2O-0.1% TFA) to give the TFA
salt of the title compound
(R)-2-(dimethylamino)-2-(2-chlorophenyl)acetic acid as a colorless
oil (0.290 g, 55%). .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
7.59-7.65 (m, 2H), 7.45-7.53 (m, 2H), 5.40 (s, 1H), 2.87 (s, 6H).
LC-MS: Anal. Calcd. for C.sub.10H.sub.12ClNO.sub.2: 213; found: 214
(M+H).sup.+.
Cap-40
##STR00053##
[0243] To an ice-cold solution of (R)-(2-chlorophenyl)glycine (1.00
g, 5.38 mmol) and NaOH (0.862 g, 21.6 mmol) in H.sub.2O (5.5 mL)
was added methyl chloroformate (1.00 mL, 13.5 mmol) dropwise. The
mixture was allowed to stir at 0.degree. C. for 1 hour and then it
was acidified by the addition of conc. HCl (2.5 mL). The mixture
was extracted with ethyl acetate (2.times.) and the combined
organic phase was washed (H.sub.2O, brine), dried
(Na.sub.2SO.sub.4), filtered, and concentrated in vacuo to give the
title compound
(R)-2-(methoxycarbonylamino)-2-(2-chlorophenyl)acetic acid as a
yellow-orange foam (1.31 g, 96%). .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.39-7.43 (m, 2H), 7.29-7.31 (m, 2H), 5.69 (s, 1H), 3.65
(s, 3H). LC-MS: Anal. Calcd. for C.sub.10H.sub.10ClNO.sub.4: 243;
found: 244 (M+H).sup.+.
Cap-41
##STR00054##
[0245] To a suspension of 2-(2-(chloromethyl)phenyl)acetic acid
(2.00 g, 10.8 mmol) in THF (20 mL) was added morpholine (1.89 g,
21.7 mmol) and the solution was stirred at room temperature for 3
hours. The reaction mixture was then diluted with ethyl acetate and
extracted with H.sub.2O (2.times.). The aqueous phase was
lyophilized and the residue was purified by silica gel
chromatography (BIOTAGE.RTM./0-10% methanol-CH.sub.2Cl.sub.2) to
give the title compound 2-(2-(Morpholinomethyl)phenyl)acetic acid
as a colorless solid (2.22 g, 87%). .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.37-7.44 (m, 3H), 7.29-7.33 (m, 1H), 4.24 (s,
2H), 3.83 (br s, 4H), 3.68 (s, 2H), 3.14 (br s, 4H). LC-MS: Anal.
Calcd. for C.sub.13H.sub.12NO.sub.3: 235; found: 236
(M+H).sup.+.
[0246] The following examples were similarly prepared using the
method described for Cap-41:
TABLE-US-00005 Cap-42 ##STR00055## LC-MS: Anal. Calcd. for
C.sub.14H.sub.19NO.sub.2: 233; found: 234 (M + H).sup.+. Cap-43
##STR00056## LC-MS: Anal. Calcd. for C.sub.13H.sub.17NO.sub.2: 219;
found: 220 (M + H).sup.+. Cap-44 ##STR00057## LC-MS: Anal. Calcd.
for C.sub.11H.sub.15NO.sub.2: 193; found: 194 (M + H).sup.+. Cap-45
##STR00058## LC-MS: Anal. Calcd. for
C.sub.14H.sub.20N.sub.2O.sub.2: 248; found: 249 (M + H).sup.+.
Cap-45a
##STR00059##
[0248] HMDS (1.85 mL, 8.77 mmol) was added to a suspension of
(R)-2-amino-2-phenylacetic acid p-toluenesulfonate (2.83 g, 8.77
mmol) in CH.sub.2Cl.sub.2 (10 mL) and the mixture was stirred at
room temperature for 30 minutes. Methyl isocyanate (0.5 g, 8.77
mmol) was added in one portion stirring continued for 30 minutes.
The reaction was quenched by addition of H.sub.2O (5 mL) and the
resulting precipitate was filtered, washed with H.sub.2O and
n-hexanes, and dried under vacuum.
(R)-2-(3-methylureido)-2-phenylacetic acid (1.5 g; 82%) was
recovered as a white solid and it was used without further
purification. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 2.54
(d, J=4.88 Hz, 3H) 5.17 (d, J=7.93 Hz, 1H) 5.95 (q, J=4.48 Hz, 1H)
6.66 (d, J=7.93 Hz, 1H) 7.26-7.38 (m, 5H) 12.67 (s, 1H). LC-MS:
Anal. Calcd. for C.sub.10H.sub.12N.sub.2O.sub.3 208.08 found
209.121 (M+H).sup.+; HPLC PHENOMENEX.RTM. C-18 3.0.times.46 mm, 0
to 100% B over 2 minutes, 1 minute hold time, A=90% water, 10%
methanol, 0.1% TFA, B=10% water, 90% methanol, 0.1%
[0249] TFA, RT=1.38 min, 90% homogeneity index.
Cap-46
##STR00060##
[0251] The desired product was prepared according to the method
described for Cap-45a. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
ppm 0.96 (t, J=7.17 Hz, 3H) 2.94-3.05 (m, 2H) 5.17 (d, J=7.93 Hz,
1H) 6.05 (t, J=5.19 Hz, 1H) 6.60 (d, J=7.63 Hz, 1H) 7.26-7.38 (m,
5H) 12.68 (s, 1H). LC-MS: Anal. Calcd. for
C.sub.11H.sub.14N.sub.2O.sub.3 222.10 found 223.15 (M+H).sup.+.
HPLC XTERRA.RTM. C-18 3.0.times.506 mm, 0 to 100% B over 2 minutes,
1 minute hold time, A=90% water, 10% methanol, 0.2%
H.sub.3PO.sub.4, B=10% water, 90% methanol, 0.2% H.sub.3PO.sub.4,
RT=0.87 min, 90% homogeneity index.
Cap-47
##STR00061##
[0253] Step 1: (R)-tert-Butyl
2-(3,3-dimethylureido)-2-phenylacetate. To a stirred solution of
(R)-tert-butyl-2-amino-2-phenylacetate (1.0 g, 4.10 mmol) and
Hunig's base (1.79 mL, 10.25 mmol) in DMF (40 mL) was added
dimethylcarbamoyl chloride (0.38 mL, 4.18 mmol) dropwise over 10
minutes. After stirring at room temperature for 3 hours, the
reaction was concentrated under reduced pressure and the resulting
residue was dissolved in ethyl acetate. The organic layer was
washed with H.sub.2O, 1N aq. HCl and brine, dried (MgSO.sub.4),
filtered and concentrated under reduced pressure. (R)-tert-butyl
2-(3,3-dimethylureido)-2-phenylacetate was obtained as a white
solid (0.86 g; 75%) and used without further purification. .sup.1H
NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 1.33 (s, 9H) 2.82 (s, 6H)
5.17 (d, J=7.63 Hz, 1H) 6.55 (d, J=7.32 Hz, 1H) 7.24-7.41 (m, 5H).
LC-MS: Anal. Calcd. for C.sub.15H.sub.22N.sub.2O.sub.3 278.16 found
279.23 (M+H).sup.+; HPLC PHENOMENEX.RTM. LUNA C-18 4.6.times.50 mm,
0 to 100% B over 4 minutes, 1 minute hold time, A=90% water, 10%
methanol, 0.1% TFA, B=10% water, 90% methanol, 0.1% TFA, RT=2.26
min, 97% homogeneity index.
[0254] Step 2: (R)-2-(3,3-Dimethylureido)-2-phenylacetic acid. To a
stirred solution of ((R)-tert-butyl
2-(3,3-dimethylureido)-2-phenylacetate (0.86 g, 3.10 mmol) in
CH.sub.2Cl.sub.2 (250 mL) was added TFA (15 mL) dropwise and the
resulting solution was stirred at rt for 3 hours. The desired
compound was then precipitated out of solution with a mixture of
EtOAC:Hexanes (5:20), filtered off and dried under reduced
pressure. (R)-2-(3,3-dimethylureido)-2-phenylacetic acid was
isolated as a white solid (0.59 g, 86%) and used without further
purification. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 2.82
(s, 6H) 5.22 (d, J=7.32 Hz, 1H) 6.58 (d, J=7.32 Hz, 1H) 7.28 (t,
J=7.17 Hz, 1H) 7.33 (t, J=7.32 Hz, 2H) 7.38-7.43 (m, 2H) 12.65 (s,
1H). LC-MS: Anal. Calcd. for C.sub.11H.sub.14N.sub.2O.sub.3:
222.24; found: 223.21 (M+H).sup.+. HPLC XTERRA.RTM. C-18
3.0.times.50 mm, 0 to 100% B over 2 minutes, 1 minute hold time,
A=90% water, 10% methanol, 0.2% H.sub.3PO.sub.4, B=10% water, 90%
methanol, 0.2% H.sub.3PO.sub.4, RT=0.75 min, 93% homogeneity
index.
Cap-48
##STR00062##
[0256] Step 1: (R)-tert-Butyl
2-(3-cyclopentylureido)-2-phenylacetate. To a stirred solution of
(R)-2-amino-2-phenylacetic acid hydrochloride (1.0 g, 4.10 mmol)
and Hunig's base (1.0 mL, 6.15 mmol) in DMF (15 mL) was added
cyclopentyl isocyanate (0.46 mL, 4.10 mmol) dropwise and over 10
minutes. After stirring at room temperature for 3 hours, the
reaction was concentrated under reduced pressure and the resulting
residue was taken up in ethyl acetate. The organic layer was washed
with H.sub.2O and brine, dried (MgSO.sub.4), filtered, and
concentrated under reduced pressure. (R)-tert-butyl
2-(3-cyclopentylureido)-2-phenylacetate was obtained as an opaque
oil (1.32 g; 100%) and used without further purification. .sup.1H
NMR (500 MHz, CD.sub.3Cl-D) .delta. ppm 1.50-1.57 (m, 2H) 1.58-1.66
(m, 2H) 1.87-1.97 (m, 2H) 3.89-3.98 (m, 1H) 5.37 (s, 1H) 7.26-7.38
(m, 5H). LC-MS: Anal. Calcd. for C.sub.18H.sub.26N.sub.2O.sub.3
318.19 found 319.21 (M+H).sup.+; HPLC XTERRA.RTM. C-18 3.0.times.50
mm, 0 to 100% B over 4 minutes, 1 minute hold time, A=90% water,
10% methanol, 0.1% TFA, B=10% water, 90% methanol, 0.1% TFA,
RT=2.82 min, 96% homogeneity index.
[0257] Step 2: (R)-2-(3-Cyclopentylureido)-2-phenylacetic acid. To
a stirred solution of (R)-tert-butyl
2-(3-cyclopentylureido)-2-phenylacetate (1.31 g, 4.10 mmol) in
CH.sub.2Cl.sub.2 (25 mL) was added TFA (4 mL) and trietheylsilane
(1.64 mL; 10.3 mmol) dropwise, and the resulting solution was
stirred at room temperature for 6 hours. The volatile components
were removed under reduced pressure and the crude product was
recrystallized in ethyl acetate/pentanes to yield
(R)-2-(3-cyclopentylureido)-2-phenylacetic acid as a white solid
(0.69 g, 64%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
1.17-1.35 (m, 2H) 1.42-1.52 (m, 2H) 1.53-1.64 (m, 2H) 1.67-1.80 (m,
2H) 3.75-3.89 (m, 1H) 5.17 (d, J=7.93 Hz, 1H) 6.12 (d, J=7.32 Hz,
1H) 6.48 (d, J=7.93 Hz, 1H) 7.24-7.40 (m, 5H) 12.73 (s, 1H). LC-MS:
Anal. Calcd. for C.sub.14H.sub.18N.sub.2O.sub.3: 262.31; found:
263.15 (M+H).sup.+. HPLC XTERRA.RTM. C-18 3.0.times.50 mm, 0 to
100% B over 2 minutes, 1 minute hold time, A=90% water, 10%
methanol, 0.2% H.sub.3PO.sub.4, B=10% water, 90% methanol, 0.2%
H.sub.3PO.sub.4, RT=1.24 min, 100% homogeneity index.
Cap-49
##STR00063##
[0259] To a stirred solution of 2-(benzylamino)acetic acid (2.0 g,
12.1 mmol) in formic acid (91 mL) was added formaldehyde (6.94 mL,
93.2 mmol). After five hours at 70.degree. C., the reaction mixture
was concentrated under reduced pressure to 20 mL and a white solid
precipitated. Following filtration, the mother liquors were
collected and further concentrated under reduced pressure providing
the crude product. Purification by reverse-phase preparative HPLC
(XTERRA.RTM. 30.times.100 mm, detection at 220 nm, flow rate 35
mL/min, 0 to 35% B over 8 min; A=90% water, 10% methanol, 0.1% TFA,
B=10% water, 90% methanol, 0.1% TFA) provided the title compound
2-(benzyl(methyl)-amino)acetic acid as its TFA salt (723 mg, 33%)
as a colorless wax. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm
2.75 (s, 3H) 4.04 (s, 2H) 4.34 (s, 2H) 7.29-7.68 (m, 5H). LC-MS:
Anal. Calcd. for: C.sub.10H.sub.13NO.sub.2 179.09; found: 180.20
(M+H).sup.+.
Cap-50
##STR00064##
[0261] To a stirred solution of 3-methyl-2-(methylamino)butanoic
acid (0.50 g, 3.81 mmol) in water (30 mL) was added K.sub.2CO.sub.3
(2.63 g, 19.1 mmol) and benzyl chloride (1.32 g, 11.4 mmol). The
reaction mixture was stirred at ambient temperature for 18 hours.
The reaction mixture was extracted with ethyl acetate (30
mL.times.2) and the aqueous layer was concentrated under reduced
pressure providing the crude product which was purified by
reverse-phase preparative HPLC (XTERRA.RTM. 30.times.100 mm,
detection at 220 nm, flow rate 40 mL/min, 20 to 80% B over 6 min;
A=90% water, 10% methanol, 0.1% TFA, B=10% water, 90% methanol,
0.1% TFA) to provide 2-(benzyl(methyl)amino)-3-methylbutanoic acid,
TFA salt (126 mg, 19%) as a colorless wax. .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 0.98 (d, 3H) 1.07 (d, 3H) 2.33-2.48 (m,
1H) 2.54-2.78 (m, 3H) 3.69 (s, 1H) 4.24 (s, 2H) 7.29-7.65 (m, 5H).
LC-MS: Anal. Calcd. for: C.sub.13H.sub.19NO.sub.2 221.14; found:
222.28 (M+H).sup.+.
Cap-51
##STR00065##
[0262] (S)-2-(Methoxycarbonylamino)-3-methylbutanoic acid
[0263] Na.sub.2CO.sub.3 (1.83 g, 17.2 mmol) was added to NaOH (33
mL of 1M/H.sub.2O, 33 mmol) solution of L-valine (3.9 g, 33.29
mmol) and the resulting solution was cooled with ice-water bath.
Methyl chloroformate (2.8 mL, 36.1 mmol) was added dropwise over 15
min, the cooling bath was removed and the reaction mixture was
stirred at ambient temperature for 3.25 hr. The reaction mixture
was washed with ether (50 mL, 3.times.), and the aqueous phase was
cooled with ice-water bath and acidified with concentrated HCl to a
pH region of 1-2, and extracted with CH.sub.2Cl.sub.2 (50 mL,
3.times.). The organic phase was dried (MgSO.sub.4) and evaporated
in vacuo to afford Cap-51 as a white solid (6 g). .sup.1H NMR for
the dominant rotamer (DMSO-d.sub.6, .delta.=2.5 ppm, 500 MHz):
12.54 (s, 1H), 7.33 (d, J=8.6, 1H), 3.84 (dd, J=8.4, 6.0, 1H), 3.54
(s, 3H), 2.03 (m, 1H), 0.87 (m, 6H). HRMS: Anal. Calcd. for
[M+H].sup.+ C.sub.21H.sub.14NO.sub.4: 176.0923; found 176.0922.
Cap-51 (Alternate Route)
##STR00066##
[0264] (S)-2-(Methoxycarbonylamino)-3-methylbutanoic acid
[0265] DIEA (137.5 mL, 0.766 mol) was added to a suspension of
(S)-tert-butyl 2-amino-3-methylbutanoate hydrochloride (75.0 g,
0.357 mol) in THF (900 mL), and the mixture was cooled to 0.degree.
C. (ice/water bath). Methyl chloroformate (29.0 mL, 0.375 mol) was
added dropwise over 45 min, the cooling bath was removed and the
heterogeneous mixture was stirred at ambient temperature for 3 h.
The solvent was removed under diminished pressure and the residue
partitioned between EtOAc and water (1 L each). The organic layer
was washed with H.sub.2O (1 L) and brine (1 L), dried (MgSO.sub.4),
filtered and concentrated under diminished pressure. The crude
material was passed through a plug of silica gel (1 kg), eluting
with hexanes (4 L) and 15:85 EtOAc/hexanes (4 L) to afford
(S)-tert-butyl 2-(methoxycarbonylamino)-3-methylbutanoate as a
clear oil (82.0 g, 99% yield). .sup.1H NMR (500 MHz, DMSO-d.sub.6,
.delta.=2.5 ppm) 7.34 (d, J=8.6, 1H), 3.77 (dd, J=8.6, 6.1, 1H),
3.53 (s, 3H), 1.94-2.05 (m, 1H), 1.39 (s, 9H), 0.83-0.92 (m, 6H).
.sup.13C-NMR (126 MHz, DMSO-d.sub.6, .delta.=39.2 ppm) 170.92,
156.84, 80.38, 60.00, 51.34, 29.76, 27.62, 18.92, 17.95. LC-MS:
[M+Na].sup.+254.17.
[0266] Trifluoroacetic acid (343 mL, 4.62 mol) and Et.sub.3SiH (142
mL, 0.887 mol) were added sequentially to a solution of
(S)-tert-butyl 2-(methoxycarbonylamino)-3-methylbutanoate (82.0 g,
0.355 mol) in CH.sub.2Cl.sub.2 (675 mL), and the mixture was
stirred at ambient temperature for 4 h. The volatile component was
removed under diminished pressure and the resultant oil triturated
with petroleum ether (600 mL) to afford a white solid, which was
filtered and washed with hexanes (500 mL) and petroleum ether (500
mL). Recrystallization from EtOAc/petroleum ether afforded Cap-51
as white flaky crystals (54.8 g, 88% yield). MP=108.5-109.5.degree.
C. .sup.1H NMR (500 MHz, DMSO-d.sub.6, .delta.=2.5 ppm) 12.52 (s,
1H), 7.31 (d, J=8.6, 1H), 3.83 (dd, J=8.6, 6.1, 1H), 3.53 (s, 3H),
1.94-2.07 (m, 1H), 0.86 (dd, J=8.9, 7.0, 6H). .sup.13C NMR (126
MHz, DMSO-d.sub.6, .delta.=39.2 ppm) 173.30, 156.94, 59.48, 51.37,
29.52, 19.15, 17.98. LC-MS: [M+H].sup.+=176.11.
[0267] Anal. Calcd. for C.sub.2H.sub.13NO.sub.4: C, 47.99; H, 7.48;
N, 7.99. Found: C, 48.17; H, 7.55; N, 7.99. Optical Rotation:
[.alpha.].sub.D=-4.16 (12.02 mg/mL; MeOH). Optical purity:
>99.5% ee. Note: the optical purity assessment was made on the
methyl ester derivative of Cap-51, which was prepared under a
standard TMSCHN.sub.2 (benzene/MeOH) esterification protocol. HPLC
analytical conditions: column, CHIRALPAK.RTM. AD-H (4.6.times.250
mm, 5 .mu.m); solvent, 95% heptane/5% IPA (isocratic); flow rate, 1
mL/min; temperature, 35.degree. C.; UV monitored at 205 nm.
[Note: Cap-51 could also be purchased from Flamm.]
Cap-52 (Same as Cap-12)
##STR00067##
[0268] (S)-2-(Methoxycarbonylamino)propanoic acid
[0269] Cap-52 was synthesized from L-alanine according to the
procedure described for the synthesis of Cap-51. For
characterization purposes, a portion of the crude material was
purified by a reverse phase HPLC(H.sub.2O/methanol/TFA) to afford
Cap-52 as a colorless viscous oil. .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5 ppm, 500 MHz): 12.49 (br s, 1H), 7.43 (d, J=7.3,
0.88H), 7.09 (app br s, 0.12H), 3.97 (m, 1H), 3.53 (s, 3H), 1.25
(d, J=7.3, 3H).
Cap-53 to Cap-64
[0270] Cap-53 to Cap-64 were prepared from appropriate starting
materials according to the procedure described for the synthesis of
Cap-51, with noted modifications if any.
TABLE-US-00006 Cap Structure Data Cap-53a: (R) Cap-53b: (S) ((S)-2-
(methoxy-carbonyl- amino)butanoic acid) ##STR00068## .sup.1H NMR
(DMSO-d.sub.6, .delta. = 2.5 ppm, 500 MHz): .delta. 12.51 (br s,
1H), 7.4 (d, J = 7.9, 0.9H), 7.06 (app s, 0.1H), 3.86-3.82 (m, 1H),
3.53 (s, 3H), 1.75-1.67 (m, 1H), 1.62- 1.54 (m, 1H), 0.88 (d, J =
7.3, 3H). RT = 0.77 minutes (Cond. 2); LC-MS: Anal. Calcd. for [M +
Na].sup.+ C.sub.6H.sub.11NNaO.sub.4: 184.06; found 184.07. HRMS
Calcd. for [M + Na].sup.+ C.sub.6H.sub.11NNaO.sub.4: 184.0586;
found 184.0592. Cap-54a: (R) Cap-54b: (S) ((S)-2- cyclopropyl-2-
(methoxy-carbonyl- amino)acetic acid) ##STR00069## .sup.1H NMR
(DMSO-d.sub.6, .delta. = 2.5 ppm, 500 MHz): .delta. 12.48 (s, 1H),
7.58 (d, J = 7.6, 0.9H), 7.25 (app s, 0.1H), 3.52 (s, 3H),
3.36-3.33 (m, 1H), 1.10-1.01 (m, 1H), 0.54- 0.49 (m, 1H), 0.46-0.40
(m, 1H), 0.39-0.35 (m, 1H), 0.31-0.21 (m, 1H). HRMS Calcd. for [M +
H].sup.+ C.sub.7H.sub.12NO.sub.4: 174.0766; found 174.0771 Cap-55
##STR00070## .sup.1H NMR (DMSO-d.sub.6, .delta. = 2.5 ppm, 500
MHz): .delta. 12.62 (s, 1H), 7.42 (d, J = 8.2, 0.9H), 7.07 (app s,
0.1H), 5.80-5.72 (m, 1H), 5.10 (d, J = 17.1, 1H), 5.04 (d, J =
10.4, 1H), 4.01-3.96 (m, 1H), 3.53 (s, 3H), 2.47-2.42 (m, 1H),
2.35-2.29 (m, 1H). Cap-56 (S)-3-methoxy-2- (methoxy-carbonyl-
amino)propanoic acid ##STR00071## .sup.1H NMR (DMSO-d.sub.6,
.delta. = 2.5 ppm, 500 MHz): .delta. 12.75 (s, 1H), 7.38 (d, J =
8.3, 0.9H), 6.96 (app s, 0.1H), 4.20-4.16 (m, 1H), 3.60-3.55 (m,
2H), 3.54 (s, 3H), 3.24 (s, 3H). Cap-57 ##STR00072## .sup.1H NMR
(DMSO-d.sub.6, .delta. = 2.5 ppm, 500 MHz): .delta. 12.50 (s, 1H),
8.02 (d, J = 7.7, 0.08H), 7.40 (d, J = 7.9, 0.76H), 7.19 (d, J =
8.2, 0.07H), 7.07 (d, J = 6.7, 0.09H), 4.21-4.12 (m, 0.08H),
4.06-3.97 (m, 0.07H), 3.96-3.80 (m, 0.85H), 3.53 (s, 3H), 1.69-
1.51 (m, 2H), 1.39-1.26 (m, 2H), 0.85 (t, J = 7.4, 3H). LC (Cond.
2): RT = 1.39 LC- MS: Anal. Calcd. for [M + H].sup.+
C.sub.7H.sub.14NO.sub.4: 176.09; found 176.06. Cap-58 ##STR00073##
.sup.1H NMR (DMSO-d.sub.6, .delta. = 2.5 ppm, 500 MHz): .delta.
12.63 (br s, 1H), 7.35 (s,1H), 7.31 (d, J = 8.2, 1H), 6.92 (s, 1H),
4.33-4.29 (m, 1H), 3.54 (s, 3H), 2.54(dd, J = 15.5, 5.4, 1H), 2.43
(dd, J = 15.6, 8.0, 1H). RT = 0.16 min (Cond. 2); LC-MS: Anal.
Calcd. for [M + H].sup.+ C.sub.6H.sub.11N.sub.2O.sub.5: 191.07;
found 191.14. Cap-59a: (R) Cap-59b: (S) ##STR00074## .sup.1H NMR
(DMSO-d.sub.6, .delta. = 2.5 ppm, 400 MHz): .delta. 12.49 (br s,
1H), 7.40 (d, J = 7.3, 0.89H), 7.04 (br s, 0.11H), 4.00-3.95 (m,
3H), 1.24 (d, J = 7.3, 3H), 1.15 (t, J = 7.2, 3H). HRMS: Anal.
Calcd. for [M + H].sup.+ C.sub.6H.sub.12NO.sub.4: 162.0766; found
162.0771. Cap-60 ##STR00075## The crude material was purified with
a reverse phase HPLC (H.sub.2O/MeOH/TFA) to afford a colorless
viscous oil that crystallized to a white solid upon exposure to
high vacuum. .sup.1H NMR (DMSO-d.sub.6, .delta. = 2.5 ppm, 400
MHz): .delta. 12.38 (br s, 1H), 7.74 (s, 0.82H), 7.48 (s, 0.18H),
3.54/3.51 (two s, 3H), 1.30 (m, 2H), 0.98 (m, 2H). HRMS: Anal.
Calcd. for [M + H].sup.+ C.sub.6H.sub.10NO.sub.4: 160.0610; found
160.0604. Cap-61 ##STR00076## .sup.1H NMR (DMSO-d.sub.6, .delta. =
2.5 ppm, 400 MHz): .delta. 12.27 (br s, 1H), 7.40 (br s, 1H), 3.50
(s, 3H), 1.32 (s, 6H). HRMS: Anal. Calcd. for [M + H].sup.+
C.sub.6H.sub.12NO.sub.4: 162.0766; found 162.0765. Cap-62
##STR00077## .sup.1H NMR (DMSO-d.sub.6, .delta. = 2.5 ppm, 400
MHz): .delta. 12.74 (br s, 1H), 4.21 (d, J = 10.3, 0.6H), 4.05 (d,
J = 10.0, 0.4H), 3.62/3.60 (two singlets, 3H), 3.0 (s, 3H),
2.14-2.05 (m, 1H), 0.95 (d, J = 6.3, 3H), 0.81 (d, J = 6.6, 3H).
LC-MS: Anal. Calcd. for [M-H].sup.- C.sub.8H.sub.14NO.sub.4:
188.09; found 188.05. Cap-63 ##STR00078## [Note: the reaction was
allowed to run for longer than what was noted for the general
procedure.] .sup.1H NMR (DMSO-d.sub.6, .delta. = 2.5 ppm, 400 MHz):
12.21 (br s, 1H), 7.42 (br s, 1H), 3.50 (s, 3H), 2.02-1.85 (m, 4H),
1.66-1.58 (m, 4H). LC-MS: Anal. Calcd. for [M + H].sup.+
C.sub.8H.sub.14NO.sub.4: 188.09; found 188.19. Cap-64 ##STR00079##
[Note: the reaction was allowed to run for longer than what was
noted for the general procedure.] .sup.1H NMR (DMSO-d.sub.6,
.delta. = 2.5 ppm, 400 MHz): 12.35 (br s, 1H), 7.77 (s, 0.82H),
7.56/7.52 (overlapping br s, 0.18H), 3.50 (s, 3H), 2.47-2.40 (m,
2H), 2.14-2.07 (m, 2H), 1.93-1.82 (m, 2H).
Cap-65
##STR00080##
[0272] Methyl chloroformate (0.65 mL, 8.39 mmol) was added dropwise
over 5 min to a cooled (ice-water) mixture of Na.sub.2CO.sub.3
(0.449 g, 4.23 mmol), NaOH (8.2 mL of 1M/H.sub.2O, 8.2 mmol) and
(S)-2-amino-3-hydroxy-3-methylbutanoic acid (1.04 g, 7.81 mmol).
The reaction mixture was stirred for 45 min, and then the cooling
bath was removed and stirring was continued for an additional 3.75
hr. The reaction mixture was washed with CH.sub.2Cl.sub.2, and the
aqueous phase was cooled with ice-water bath and acidified with
concentrated HCl to a pH region of 1-2. The volatile component was
removed in vacuo and the residue was taken up in a 2:1 mixture of
MeOH/CH.sub.2Cl.sub.2 (15 mL) and filtered, and the filterate was
rotervaped to afford Cap-65 as a white semi-viscous foam (1.236 g).
.sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): .delta. 6.94
(d, J=8.5, 0.9H), 6.53 (br s, 0.1H), 3.89 (d, J=8.8, 1H), 2.94 (s,
3H), 1.15 (s, 3H), 1.13 (s, 3H).
[0273] Cap-66 and Cap-67 were prepared from appropriate
commercially available starting materials by employing the
procedure described for the synthesis of Cap-65.
Cap-66
##STR00081##
[0275] .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz):
.delta. 12.58 (br s, 1H), 7.07 (d, J=8.3, 0.13H), 6.81 (d, J=8.8,
0.67H), 4.10-4.02 (m, 1.15H), 3.91 (dd, J=9.1, 3.5, 0.85H), 3.56
(s, 3H), 1.09 (d, J=6.2, 3H). [Note: only the dominant signals of
NH were noted].
Cap-67
##STR00082##
[0277] .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): 12.51
(br s, 1H), 7.25 (d, J=8.4, 0.75H), 7.12 (br d, J=0.4, 0.05H), 6.86
(br s, 0.08H), 3.95-3.85 (m, 2H), 3.54 (s, 3H), 1.08 (d, J=6.3,
3H). [Note: only the dominant signals of NH were noted].
Cap-68
##STR00083##
[0279] Methyl chloroformate (0.38 ml, 4.9 mmol) was added drop-wise
to a mixture of 1N NaOH (aq) (9.0 ml, 9.0 mmol), 1M NaHCO.sub.3
(aq) (9.0 ml, 9.0 mol), L-aspartic acid .beta.-benzyl ester (1.0 g,
4.5 mmol) and Dioxane (9 ml). The reaction mixture was stirred at
ambient conditions for 3 hr, and then washed with Ethyl acetate (50
ml, 3.times.). The aqueous layer was acidified with 12N HCl to a pH
.about.1-2, and extracted with ethyl acetate (3.times.50 ml). The
combined organic layers were washed with brine, dried
(Na.sub.2SO.sub.4), filtered, and concentrated in vacuo to afford
Cap-68 as a light yellow oil (1.37 g; mass is above theoretical
yield, and the product was used without further purification).
.sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 500 MHz): .delta. 12.88
(br s, 1H), 7.55 (d, J=8.5, 1H), 7.40-7.32 (m, 5H), 5.13 (d,
J=12.8, 1H), 5.10 (d, J=12.9, 1H), 4.42-4.38 (m, 1H), 3.55 (s, 3H),
2.87 (dd, J=16.2, 5.5, 1H), 2.71 (dd, J=16.2, 8.3, 1H). LC (Cond.
2): RT=1.90 min; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.13H.sub.16NO.sub.6: 282.10; found 282.12.
Cap-69a and Cap-69b
##STR00084##
[0281] NaCNBH.sub.3 (2.416 g, 36.5 mmol) was added in batches to a
chilled (.about.15.degree. C.) water (17 mL)/MeOH (10 mL) solution
of alanine (1.338 g, 15.0 mmol). A few minutes later acetaldehyde
(4.0 mL, 71.3 mmol) was added drop-wise over 4 min, the cooling
bath was removed, and the reaction mixture was stirred at ambient
condition for 6 hr. An additional acetaldehyde (4.0 mL) was added
and the reaction was stirred for 2 hr.
[0282] Concentrated HCl was added slowly to the reaction mixture
until the pH reached .about.1.5, and the resulting mixture was
heated for 1 hr at 40.degree. C. Most of the volatile component was
removed in vacuo and the residue was purified with a DOWEX.RTM.
50WX8-100 ion-exchange resin (column was washed with water, and the
compound was eluted with dilute NH.sub.4OH, prepared by mixing 18
ml of NH.sub.4OH and 282 ml of water) to afford Cap-69 (2.0 g) as
an off-white soft hygroscopic solid. .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5 ppm, 400 MHz): .delta. 3.44 (q, J=7.1, 1H), 2.99-2.90
(m, 2H), 2.89-2.80 (m, 2H), 1.23 (d, J=7.1, 3H), 1.13 (t, J=7.3,
6H).
Cap-70 to Cap-74x
[0283] Cap-70 to Cap-74x were prepared according to the procedure
described for the synthesis of Cap-69 by employing appropriate
starting materials.
TABLE-US-00007 Cap-70a: (R) Cap-70b: (S) ##STR00085## .sup.1H NMR
(DMSO-d.sub.6, .delta. = 2.5 ppm, 400 MHz): .delta. 3.42 (q, J =
7.1, 1H), 2.68-2.60 (m, 4H), 1.53-1.44 (m, 4H), 1.19 (d, J = 7.3,
3H), 0.85 (t, J = 7.5, 6H). LC- MS: Anal. Calcd. for [M + H].sup.+
C.sub.9H.sub.20NO.sub.2: 174.15; found 174.13. Cap-71a: (R)
Cap-71b: (S) ##STR00086## .sup.1H NMR (DMSO-d.sub.6, .delta. = 2.5
ppm, 500 MHz): .delta. 3.18- 3.14 (m, 1H), 2.84-2.77 (m, 2H),
2.76-2.68 (m, 2H), 1.69-1.54 (m, 2H), 1.05 (t, J = 7.2, 6H), 0.91
(t, J = 7.3, 3H). LC-MS: Anal. Calcd. for [M + H].sup.+
C.sub.8H.sub.18NO.sub.2: 160.13; found 160.06. Cap-72 ##STR00087##
.sup.1H NMR (DMSO-d.sub.6, .delta. = 2.5 ppm, 400 MHz): .delta.
2.77- 2.66 (m, 3H), 2.39-2.31 (m, 2H), 1.94-1.85 (m, 1H), 0.98 (t,
J = 7.1, 6H), 0.91 (d, J = 6.5, 3H), 0.85 (d, J = 6.5, 3H). LC-MS:
Anal. Calcd. for [M + H].sup.+ C.sub.9H.sub.20NO.sub.2: 174.15;
found 174.15. Cap-73 ##STR00088## .sup.1H NMR (DMSO-d.sub.6,
.delta. = 2.5 ppm, 500 MHz): .delta. 9.5 (br s, 1H), 3.77 (dd, J =
10.8, 4.1, 1H), 3.69-3.61 (m, 2H), 3.26 (s, 3H), 2.99-2.88 (m, 4H),
1.13 (t, J = 7.2, 6H). Cap-74 ##STR00089## .sup.1H NMR
(DMSO-d.sub.6, .delta. = 2.5 ppm, 500 MHz): .delta. 7.54 (s, 1H),
6.89 (s, 1H), 3.81 (t, J = 6.6, k, 1H), 2.82- 2.71 (m, 4H), 2.63
(dd, J = 15.6, 7.0, 1H), 2.36 (dd, J = 15.4, 6.3, 1H), 1.09 (t, J =
7.2, 6H). RT = 0.125 minutes (Cond. 2); LC-MS: Anal. Calcd. for [M
+ H].sup.+ C.sub.8H.sub.17N.sub.2O.sub.3: 189.12; found 189.13.
Cap-74x ##STR00090## LC-MS: Anal. Calcd. for [M + H].sup.+
C.sub.10H.sub.22NO.sub.2: 188.17; found 188.21
Cap-75
##STR00091##
[0284] Cap-75, Step a
##STR00092##
[0286] NaBH.sub.3CN (1.6 g, 25.5 mmol) was added to a cooled
(ice/water bath) water (25 ml)/methanol (15 ml) solution of
H-D-Ser-OBzl HCl (2.0 g, 8.6 mmol). Acetaldehyde (1.5 ml, 12.5
mmol) was added drop-wise over 5 min, the cooling bath was removed,
and the reaction mixture was stirred at ambient condition for 2 hr.
The reaction was carefully quenched with 12N HCl and concentrated
in vacuo. The residue was dissolved in water and purified with a
reverse phase HPLC (MeOH/H.sub.2O/TFA) to afford the TFA salt of
(R)-benzyl 2-(diethylamino)-3-hydroxypropanoate as a colorless
viscous oil (1.9 g). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm,
500 MHz): .delta. 9.73 (br s, 1H), 7.52-7.36 (m, 5H), 5.32 (d,
J=12.2, 1H), 5.27 (d, J=12.5, 1H), 4.54-4.32 (m, 1H), 4.05-3.97 (m,
2H), 3.43-3.21 (m, 4H), 1.23 (t, J=7.2, 6H). LC-MS (Cond. 2):
RT=1.38 min; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.14H.sub.22NO.sub.3: 252.16; found 252.19.
Cap-75
[0287] NaH (0.0727 g, 1.82 mmol, 60%) was added to a cooled
(ice-water) THF (3.0 mL) solution of the TFA salt (R)-benzyl
2-(diethylamino)-3-hydroxypropanoate (0.3019 g, 0.8264 mmol)
prepared above, and the mixture was stirred for 15 min. Methyl
iodide (56 .mu.L, 0.90 mmol) was added and stirring was continued
for 18 hr while allowing the bath to thaw to ambient condition. The
reaction was quenched with water and loaded onto a MeOH
pre-conditioned MCX (6 g) cartridge, and washed with methanol
followed by compound elution with 2N NH.sub.3/Methanol. Removal of
the volatile component in vacuo afforded Cap-75, contaminated with
(R)-2-(diethylamino)-3-hydroxypropanoic acid, as a yellow
semi-solid (100 mg). The product was used as is without further
purification.
Cap-76
##STR00093##
[0289] NaCNBH.sub.3 (1.60 g, 24.2 mmol) was added in batches to a
chilled (.about.15.degree. C.) water/MeOH (12 mL each) solution of
(S)-4-amino-2-(tert-butoxycarbonylamino) butanoic acid (2.17 g,
9.94 mmol). A few minutes later acetaldehyde (2.7 mL, 48.1 mmol)
was added drop-wise over 2 min, the cooling bath was removed, and
the reaction mixture was stirred at ambient condition for 3.5 hr.
An additional acetaldehyde (2.7 mL, 48.1 mmol) was added and the
reaction was stirred for 20.5 hr. Most of the MeOH component was
removed in vacuo, and the remaining mixture was treated with
concentrated HCl until its pH reached .about.1.0 and then heated
for 2 hr at 40.degree. C. The volatile component was removed in
vacuo, and the residue was treated with 4 M HCl/dioxane (20 mL) and
stirred at ambient condition for 7.5 hr. The volatile component was
removed in vacuo and the residue was purified with DOWEX.RTM.
50WX8-100 ion-exchange resin (column was washed with water and the
compound was eluted with dilute NH.sub.4OH, prepared from 18 ml of
NH.sub.4OH and 282 ml of water) to afford intermediate
(S)-2-amino-4-(diethylamino)butanoic acid as an off-white solid
(1.73 g).
[0290] Methyl chloroformate (0.36 mL, 4.65 mmol) was added
drop-wise over 11 min to a cooled (ice-water) mixture of
Na.sub.2CO.sub.3 (0.243 g, 2.29 mmol), NaOH (4.6 mL of 1M/H.sub.2O,
4.6 mmol) and the above product (802.4 mg). The reaction mixture
was stirred for 55 min, and then the cooling bath was removed and
stirring was continued for an additional 5.25 hr. The reaction
mixture was diluted with equal volume of water and washed with
CH.sub.2Cl.sub.2 (30 mL, 2.times.), and the aqueous phase was
cooled with ice-water bath and acidified with concentrated HCl to a
pH region of 2. The volatile component was then removed in vacuo
and the crude material was free-based with MCX resin (6.0 g; column
was washed with water, and sample was eluted with 2.0 M
NH.sub.3/MeOH) to afford impure Cap-76 as an off-white solid (704
mg). .sup.1H NMR (MeOH-d.sub.4, .delta.=3.29 ppm, 400 MHz): .delta.
3.99 (dd, J=7.5, 4.7, 1H), 3.62 (s, 3H), 3.25-3.06 (m, 6H),
2.18-2.09 (m, 1H), 2.04-1.96 (m, 1H), 1.28 (t, J=7.3, 6H). LC-MS:
Anal. Calcd. for [M+H].sup.+ C.sub.10H.sub.21N.sub.2O.sub.4:
233.15; found 233.24.
Cap-77a and Cap-77b
##STR00094##
[0292] The synthesis of Cap-77 was conducted according to the
procedure described for Cap-7 by using 7-azabicyclo[2.2.1]heptane
for the SN.sub.2 displacement step, and by effecting the
stereoisomeric separation of the intermediate benzyl
2-(7-azabicyclo[2.2.1]heptan-7-yl)-2-phenylacetate using the
following condition: the intermediate (303.7 mg) was dissolved in
ethanol, and the resulting solution was injected on a chiral HPLC
column (Chiracel AD-H column, 30.times.250 mm, 5 um) eluting with
90% CO.sub.2-10% EtOH at 70 mL/min, and a temperature of 35.degree.
C. to provide 124.5 mg of stereoisomer-1 and 133.8 mg of
stereoisomer-2. These benzyl esters were hydrogenolysed according
to the preparation of Cap-7 to provide Cap-77: .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): .delta. 7.55 (m, 2H),
7.38-7.30 (m, 3H), 4.16 (s, 1H), 3.54 (app br s, 2H), 2.08-1.88 (m,
4H), 1.57-1.46 (m, 4H). LC (Cond. 1): RT=0.67 min; LC-MS: Anal.
Calcd. for [M+H].sup.+ C.sub.14H.sub.18NO.sub.2: 232.13; found
232.18. HRMS: Anal. Calcd. for [M+H].sup.+
C.sub.14H.sub.18NO.sub.2: 232.1338; found 232.1340.
Cap-78
##STR00095##
[0294] NaCNBH.sub.3 (0.5828 g, 9.27 mmol) was added to a mixture of
the HCl salt of (R)-2-(ethylamino)-2-phenylacetic acid (an
intermediate in the synthesis of Cap-3; 0.9923 mg, 4.60 mmol) and
(1-ethoxycyclopropoxy)trimethylsilane (1.640 g, 9.40 mmol) in MeOH
(10 mL), and the semi-heterogeneous mixture was heated at
50.degree. C. with an oil bath for 20 hr. More
(1-ethoxycyclopropoxy)trimethylsilane (150 mg, 0.86 mmol) and
NaCNBH.sub.3 (52 mg, 0.827 mmol) were added and the reaction
mixture was heated for an additional 3.5 hr. It was then allowed to
cool to ambient temperature and acidified to a .about.pH region of
2 with concentrated HCl, and the mixture was filtered and the
filtrate was rotervaped. The resulting crude material was taken up
in i-PrOH (6 mL) and heated to effect dissolution, and the
non-dissolved part was filtered off and the filtrate concentrated
in vacuo. About 1/3 of the resultant crude material was purified
with a reverse phase HPLC (H.sub.2O/MeOH/TFA) to afford the TFA
salt of Cap-78 as a colorless viscous oil (353 mg). .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz; after D.sub.2O exchange):
.delta. 7.56-7.49 (m, 5H), 5.35 (S, 1H), 3.35 (m, 1H), 3.06 (app br
s, 1H), 2.66 (m, 1H), 1.26 (t, J=7.3, 3H), 0.92 (m, 1H), 0.83-0.44
(m, 3H). LC (Cond. 1): RT=0.64 min; LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.13H.sub.18NO.sub.2: 220.13; found 220.21. HRMS:
Anal. Calcd. for [M+H].sup.+ C.sub.13H.sub.18NO.sub.2: 220.1338;
found 220.1343.
Cap-79
##STR00096##
[0296] Ozone was bubbled through a cooled (-78.degree. C.)
CH.sub.2Cl.sub.2 (5.0 mL) solution Cap-55 (369 mg, 2.13 mmol) for
about 50 min until the reaction mixture attained a tint of blue
color. Me.sub.2S (10 pipet drops) was added, and the reaction
mixture was stirred for 35 min. The -78.degree. C. bath was
replaced with a -10.degree. C. bath and stirring continued for an
additional 30 min, and then the volatile component was removed in
vacuo to afford a colorless viscous oil.
[0297] NaBH.sub.3CN (149 mg, 2.25 mmol) was added to a MeOH (5.0
mL) solution of the above crude material and morpholine (500 .mu.L,
5.72 mmol) and the mixture was stirred at ambient condition for 4
hr. It was cooled to ice-water temperature and treated with
concentrated HCl to bring its pH to .about.2.0, and then stirred
for 2.5 hr. The volatile component was removed in vacuo, and the
residue was purified with a combination of MCX resin (MeOH wash;
2.0 N NH.sub.3/MeOH elution) and a reverse phase HPLC
(H.sub.2O/MeOH/TFA) to afford Cap-79 containing unknown amount of
morpholine.
[0298] In order to consume the morpholine contaminant, the above
material was dissolved in CH.sub.2Cl.sub.2 (1.5 mL) and treated
with Et.sub.3N (0.27 mL, 1.94 mmol) followed by acetic anhydride
(0.10 mL, 1.06 mmol) and stirred at ambient condition for 18 hr.
THF (1.0 mL) and H.sub.2O (0.5 mL) were added and stirring
continued for 1.5 hr. The volatile component was removed in vacuo,
and the resultant residue was passed through MCX resin (MeOH wash;
2.0 N NH.sub.3/MeOH elution) to afford impure Cap-79 as a brown
viscous oil, which was used for the next step without further
purification.
Cap-80a and Cap-80b
##STR00097##
[0300] SOCl.sub.2 (6.60 mL, 90.5 mmol) was added drop-wise over 15
min to a cooled (ice-water) mixture of
(S)-3-amino-4-(benzyloxy)-4-oxobutanoic acid (10.04 g, 44.98 mmol)
and MeOH (300 mL), the cooling bath was removed and the reaction
mixture was stirred at ambient condition for 29 hr. Most of the
volatile component was removed in vacuo and the residue was
carefully partitioned between EtOAc (150 mL) and saturated
NaHCO.sub.3 solution. The aqueous phase was extracted with EtOAc
(150 mL, 2.times.), and the combined organic phase was dried
(MgSO.sub.4), filtered, and concentrated in vacuo to afford
(S)-1-benzyl 4-methyl 2-aminosuccinate as a colorless oil (9.706
g). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): .delta.
7.40-7.32 (m, 5H), 5.11 (s, 2H), 3.72 (app t, J=6.6, 1H), 3.55 (s,
3H), 2.68 (dd, J=15.9, 6.3, 1H), 2.58 (dd, J=15.9, 6.8, 1H), 1.96
(s, 2H). LC (Cond. 1): RT=0.90 min; LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.12H.sub.16NO.sub.4: 238.11; found 238.22.
[0301] Pb(NO.sub.3).sub.2 (6.06 g, 18.3 mmol) was added over 1 min
to a CH.sub.2Cl.sub.2 (80 mL) solution of (S)-1-benzyl 4-methyl
2-aminosuccinate (4.50 g, 19.0 mmol), 9-bromo-9-phenyl-9H-fluorene
(6.44 g, 20.0 mmol) and Et.sub.3N (3.0 mL, 21.5 mmol), and the
heterogeneous mixture was stirred at ambient condition for 48 hr.
The mixture was filtered and the filtrate was treated with
MgSO.sub.4 and filtered again, and the final filtrate was
concentrated. The resulting crude material was submitted to a
BIOTAGE.RTM. purification (350 g silica gel, CH.sub.2Cl.sub.2
elution) to afford (S)-1-benzyl 4-methyl
2-(9-phenyl-9H-fluoren-9-ylamino)succinate as highly viscous
colorless oil (7.93 g). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm,
400 MHz): .delta. 7.82 (m, 2H), 7.39-7.13 (m, 16H), 4.71 (d,
J=12.4, 1H), 4.51 (d, J=12.6, 1H), 3.78 (d, J=9.1, NH), 3.50 (s,
3H), 2.99 (m, 1H), 2.50-2.41 (m, 2H, partially overlapped with
solvent). LC (Cond. 1): RT=2.16 min; LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.31H.sub.28NO.sub.4: 478.20; found 478.19.
[0302] LiHMDS (9.2 mL of 1.0 M/THF, 9.2 mmol) was added drop-wise
over 10 min to a cooled (-78.degree. C.) THF (50 mL) solution of
(S)-1-benzyl 4-methyl 2-(9-phenyl-9H-fluoren-9-ylamino)succinate
(3.907 g, 8.18 mmol) and stirred for .about.1 hr. Met (0.57 mL, 9.2
mmol) was added drop-wise over 8 min to the mixture, and stirring
was continued for 16.5 hr while allowing the cooling bath to thaw
to room temperature. After quenching with saturated NH.sub.4Cl
solution (5 mL), most of the organic component was removed in vacuo
and the residue was partitioned between CH.sub.2Cl.sub.2 (100 mL)
and water (40 mL). The organic layer was dried (MgSO.sub.4),
filtered, and concentrated in vacuo, and the resulting crude
material was purified with a BIOTAGE.RTM. (350 g silica gel; 25%
EtOAc/hexanes) to afford 3.65 g of a 2S/3S and 2S/3R diastereomeric
mixtures of 1-benzyl 4-methyl
3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)succinate in 1.0:0.65
ratio (.sup.1H NMR). The stereochemistry of the dominant isomer was
not determined at this juncture, and the mixture was submitted to
the next step without separation. Partial .sup.1H NMR data
(DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): major diastereomer,
.delta. 4.39 (d, J=12.3, 1H of CH.sub.2), 3.33 (s, 3H, overlapped
with H.sub.2O signal), 3.50 (d, J=10.9, NH), 1.13 (d, J=7.1, 3H);
minor diastereomer, .delta. 4.27 (d, J=12.3, 1H of CH.sub.2), 3.76
(d, J=10.9, NH), 3.64 (s, 3H), 0.77 (d, J=7.0, 3H). LC (Cond. 1):
RT=2.19 min; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.32H.sub.30NO.sub.4: 492.22; found 492.15.
[0303] Diisobutylaluminum hydride (20.57 ml of 1.0 M in hexanes,
20.57 mmol) was added drop-wise over 10 min to a cooled
(-78.degree. C.) THF (120 mL) solution of (2S)-1-benzyl 4-methyl
3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)succinate (3.37 g, 6.86
mmol) prepared above, and stirred at -78.degree. C. for 20 hr. The
reaction mixture was removed from the cooling bath and rapidly
poured into .about.1M H.sub.3PO.sub.4/H.sub.2O (250 mL) with
stirring, and the mixture was extracted with ether (100 mL,
2.times.). The combined organic phase was washed with brine, dried
(MgSO.sub.4), filtered and concentrated in vacuo. A silica gel mesh
of the crude material was prepared and submitted to chromatography
(25% EtOAc/hexanes; gravity elution) to afford 1.1 g of
(2S,3S)-benzyl
4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate,
contaminated with benzyl alcohol, as a colorless viscous oil and
(2S,3R)-benzyl
4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate
containing the (2S,3R) stereoisomer as an impurity. The later
sample was resubmitted to the same column chromatography
purification conditions to afford 750 mg of purified material as a
white foam. [Note: the (2S,3S) isomer elutes before the (2S,3R)
isomer under the above condition]. (2S,3S) isomer: .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz): 7.81 (m, 2H), 7.39-7.08
(m, 16H), 4.67 (d, J=12.3, 1H), 4.43 (d, J=12.4, 1H), 4.21 (app t,
J=5.2, OH), 3.22 (d, J=10.1, NH), 3.17 (m, 1H), 3.08 (m, 1H),
.about.2.5 (m, 1H, overlapped with the solvent signal), 1.58 (m,
1H), 0.88 (d, J=6.8, 3H). LC (Cond. 1): RT=2.00 min; LC-MS: Anal.
Calcd. for [M+H].sup.+ C.sub.31H.sub.30NO.sub.3: 464.45; found
464.22. (2S,3R) isomer: .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm,
400 MHz): 7.81 (d, J=7.5, 2H), 7.39-7.10 (m, 16H), 4.63 (d, J=12.1,
1H), 4.50 (app t, J=4.9, 1H), 4.32 (d, J=12.1, 1H), 3.59-3.53 (m,
2H), 3.23 (m, 1H), 2.44 (dd, J=9.0, 8.3, 1H), 1.70 (m, 1H), 0.57
(d, J=6.8, 3H). LC (Cond. 1): RT=1.92 min; LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.31H.sub.30NO.sub.3: 464.45; found 464.52.
[0304] The relative stereochemical assignments of the
DIBAL-reduction products were made based on NOE studies conducted
on lactone derivatives prepared from each isomer by employing the
following protocol: LiHMDS (50 .mu.L of 1.0 M/THF, 0.05 mmol) was
added to a cooled (ice-water) THF (2.0 mL) solution of
(2S,3S)-benzyl
4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate (62.7
mg, 0.135 mmol), and the reaction mixture was stirred at similar
temperature for .about.2 hr. The volatile component was removed in
vacuo and the residue was partitioned between CH.sub.2Cl.sub.2 (30
mL), water (20 mL) and saturated aqueous NH.sub.4Cl solution (1
mL). The organic layer was dried (MgSO.sub.4), filtered, and
concentrated in vacuo, and the resulting crude material was
submitted to a BIOTAGE.RTM. purification (40 g silica gel; 10-15%
EtOAc/hexanes) to afford
(3S,4S)-4-methyl-3-(9-phenyl-9H-fluoren-9-ylamino)dihydrofuran-2(3H)-one
as a colorless film of solid (28.1 mg). (2S,3R)-benzyl
4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate was
elaborated similarly to
(3S,4R)-4-methyl-3-(9-phenyl-9H-fluoren-9-ylamino)dihydrofuran-2(3H)-one.
(3S,4S)-lactone isomer: .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm,
400 MHz), 7.83 (d, J=7.5, 2H), 7.46-7.17 (m, 11H), 4.14 (app t,
J=8.3, 1H), 3.60 (d, J=5.8, NH), 3.45 (app t, J=9.2, 1H),
.about.2.47 (m, 1H, partially overlapped with solvent signal), 2.16
(m, 1H), 0.27 (d, J=6.6, 3H). LC (Cond. 1): RT=1.98 min; LC-MS:
Anal. Calcd. for [M+Na].sup.+ C.sub.24H.sub.21NNaO.sub.2: 378.15;
found 378.42. (3S,4R)-lactone isomer: .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5 ppm, 400 MHz), 7.89 (d, J=7.6, 1H), 7.85 (d, J=7.3,
1H), 7.46-7.20 (m, 11H), 3.95 (dd, J=9.1, 4.8, 1H), 3.76 (d, J=8.8,
1H), 2.96 (d, J=3.0, NH), 2.92 (dd, J=6.8, 3, NCH), 1.55 (m, 1H),
0.97 (d, J=7.0, 3H). LC (Cond. 1): RT=2.03 min; LC-MS: Anal. Calcd.
for [M+Na].sup.+ C.sub.24H.sub.21NNaO.sub.2: 378.15; found
378.49.
[0305] TBDMS-Cl (48 mg, 0.312 mmol) followed by imidazole (28.8 mg,
0.423 mmol) were added to a CH.sub.2Cl.sub.2 (3 ml) solution of
(2S,3S)-benzyl
4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate
(119.5 mg, 0.258 mmol), and the mixture was stirred at ambient
condition for 14.25 hr. The reaction mixture was then diluted with
CH.sub.2Cl.sub.2 (30 mL) and washed with water (15 mL), and the
organic layer was dried (MgSO.sub.4), filtered, and concentrated in
vacuo. The resultant crude material was purified with a
BIOTAGE.RTM. (40 g silica gel; 5% EtOAc/hexanes) to afford
(2S,3S)-benzyl
4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)-
butanoate, contaminated with TBDMS based impurities, as a colorless
viscous oil (124.4 mg). (2S,3R)-benzyl
4-hydroxy-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)butanoate was
elaborated similarly to (2S,3R)-benzyl
4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)-
butanoate. (2S,3S)-silyl ether isomer: .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5 ppm, 400 MHz), 7.82 (d, J=4.1, 1H), 7.80 (d, J=4.0,
1H), 7.38-7.07 (m, 16H), 4.70 (d, J=12.4, 1H), 4.42 (d, J=12.3,
1H), 3.28-3.19 (m, 3H), 2.56 (dd, J=10.1, 5.5, 1H), 1.61 (m, 1H),
0.90 (d, J=6.8, 3H), 0.70 (s, 9H), -0.13 (s, 3H), -0.16 (s, 3H). LC
(Cond. 1, where the run time was extended to 4 min): RT=3.26 min;
LC-MS: Anal. Calcd. for [M+H].sup.+ C.sub.37H.sub.44NO.sub.3Si:
578.31; found 578.40. (2S,3R)-silyl ether isomer: .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz), 7.82 (d, J=3.0, 1H), 7.80
(d, J=3.1, 1H), 7.39-7.10 (m, 16H), 4.66 (d, J=12.4, 1H), 4.39 (d,
J=12.4, 1H), 3.61 (dd, J=9.9, 5.6, 1H), 3.45 (d, J=9.5, 1H), 3.41
(dd, J=10, 6.2, 1H), 2.55 (dd, J=9.5, 7.3, 1H), 1.74 (m, 1H), 0.77
(s, 9H), 0.61 (d, J=7.1, 3H), -0.06 (s, 3H), -0.08 (s, 3H).
[0306] A balloon of hydrogen was attached to a mixture of
(2S,3S)-benzyl
4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)-
butanoate (836 mg, 1.447 mmol) and 10% Pd/C (213 mg) in EtOAc (16
mL) and the mixture was stirred at room temperature for .about.21
hr, where the balloon was recharged with H.sub.2 as necessary. The
reaction mixture was diluted with CH.sub.2Cl.sub.2 and filtered
through a pad of diatomaceous earth (CELITE.RTM.-545), and the pad
was washed with EtOAc (200 mL), EtOAc/MeOH (1:1 mixture, 200 mL)
and MeOH (750 mL). The combined organic phase was concentrated, and
a silica gel mesh was prepared from the resulting crude material
and submitted to a flash chromatography (8:2:1 mixture of
EtOAc/1-PrOH/H.sub.2O) to afford
(2S,3S)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic
acid as a white fluffy solid (325 mg). (2S,3R)-benzyl
4-(tert-butyldimethylsilyloxy)-3-methyl-2-(9-phenyl-9H-fluoren-9-ylamino)-
butanoate was similarly elaborated to
(2S,3R)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic
acid. (2S,3S)-amino acid isomer: .sup.1H NMR (methanol-d.sub.4,
.delta.=3.29 ppm, 400 MHz), 3.76 (dd, J=10.5, 5.2, 1H), 3.73 (d,
J=3.0, 1H), 3.67 (dd, J=10.5, 7.0, 1H), 2.37 (m, 1H), 0.97 (d,
J=7.0, 3H), 0.92 (s, 9H), 0.10 (s, 6H). LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.11H.sub.26NO.sub.3Si: 248.17; found 248.44.
(2S,3R)-amino acid isomer: .sup.1H NMR (methanol-d.sub.4,
.delta.=3.29 ppm, 400 MHz), 3.76-3.75 (m, 2H), 3.60 (d, J=4.1, 1H),
2.16 (m, 1H), 1.06 (d, J=7.3, 3H), 0.91 (s, 9H), 0.09 (s, 6H).
Anal. Calcd. for [M+H].sup.+ C.sub.11H.sub.26NO.sub.3Si: 248.17;
found 248.44.
[0307] Water (1 mL) and NaOH (0.18 mL of 1.0 M/H.sub.2O, 0.18 mmol)
were added to a mixture of
(2S,3S)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic
acid (41.9 mg, 0.169 mmol) and Na.sub.2CO.sub.3 (11.9 mg, 0.112
mmol), and sonicated for about 1 min to effect dissolution of
reactants. The mixture was then cooled with an ice-water bath,
methyl chloroformate (0.02 mL, 0.259 mmol) was added over 30 s, and
vigorous stirring was continued at similar temperature for 40 min
and then at ambient temperature for 2.7 hr. The reaction mixture
was diluted with water (5 mL), cooled with ice-water bath and
treated drop-wise with 1.0 N HCl aqueous solution (.about.0.23 mL).
The mixture was further diluted with water (10 mL) and extracted
with CH.sub.2Cl.sub.2 (15 mL, 2.times.). The combined organic phase
was dried (MgSO.sub.4), filtered, and concentrated in vacuo to
afford Cap-80a as an off-white solid.
(2S,3R)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic
acid was similarly elaborated to Cap-80b. Cap-80a: .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5 ppm, 400 MHz), 12.57 (br s, 1H), 7.64
(d, J=8.3, 0.3H), 7.19 (d, J=8.8, 0.7H), 4.44 (dd, J=8.1, 4.6,
0.3H), 4.23 (dd, J=8.7, 4.4, 0.7H), 3.56/3.53 (two singlets, 3H),
3.48-3.40 (m, 2H), 2.22-2.10 (m, 1H), 0.85 (s, 9H), .about.0.84 (d,
0.9H, overlapped with t-Bu signal), 0.79 (d, J=7, 2.1H),
0.02/0.01/0.00 (three overlapping singlets, 6H). LC-MS: Anal.
Calcd. for [M+Na].sup.+ C.sub.13H.sub.27NNaO.sub.5Si: 328.16; found
328.46. Cap-80b: .sup.1H NMR (CDCl.sub.3, .delta.=7.24 ppm, 400
MHz), 6.00 (br d, J=6.8, 1H), 4.36 (dd, J=7.1, 3.1, 1H), 3.87 (dd,
J=10.5, 3.0, 1H), 3.67 (s, 3H), 3.58 (dd, J=10.6, 4.8, 1H), 2.35
(m, 1H), 1.03 (d, J=7.1, 3H), 0.90 (s, 9H), 0.08 (s, 6H). LC-MS:
Anal. Calcd. for [M+Na].sup.+ C.sub.13H.sub.27NNaO.sub.5Si: 328.16;
found 328.53. The crude products were utilized without further
purification.
Cap-81
##STR00098##
[0309] Prepared according to the protocol described by Falb et al.,
Synthetic Communications, 23:2839 (1993).
Cap-82 to Cap-85
[0310] Cap-82 to Cap-85 were synthesized from appropriate starting
materials according to the procedure described for Cap-51 or
Cap-13. The samples exhibited similar spectral profiles as that of
their stereoisomers (i.e., Cap-4, Cap-13, Cap-51 and Cap-52,
respectively).
##STR00099##
(2S,3R)-3-Methoxy-2-(methoxycarbonylamino)butanoic acid
[0311] To a mixture of O-methyl-L-threonine (3.0 g, 22.55 mmol),
NaOH (0.902 g, 22.55 mmol) in H.sub.2O (15 mL) was added
ClCO.sub.2Me (1.74 mL, 22.55 mmol) dropwise at 0.degree. C. The
mixture was allowed to stir for 12 h and acidified to pH 1 using 1N
HCl. The aqueous phase was extracted with EtOAc and (2.times.250
mL) and 10% MeOH in CH.sub.2Cl.sub.2 (250 mL) and the combined
organic phases were concentrated under in vacuo to afford a
colorless oil (4.18 g, 97%) which was of sufficient purity for use
in subsequent steps. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.19
(s, 1H), 3.92-3.97 (m, 1H), 3.66 (s, 3H), 1.17 (d, J=7.7 Hz, 3H).
LC-MS: Anal. Calcd. for C.sub.7H.sub.13NO.sub.5: 191; found: 190
(M-H).sup.-.
Cap-87
##STR00100##
[0313] To a mixture of L-homoserine (2.0 g, 9.79 mmol),
Na.sub.2CO.sub.3 (2.08 g, 19.59 mmol) in H.sub.2O (15 mL) was added
ClCO.sub.2Me (0.76 mL, 9.79 mmol) dropwise at 0.degree. C. The
mixture was allowed to stir for 48 h and acidified to pH 1 using 1N
HCl. The aqueous phase was extracted with EtOAc and (2.times.250
mL) and the combined organic phases were concentrated in vacuo to
afford a colorless solid (0.719 g, 28%) which was of sufficient
purity for use in subsequent steps. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.23 (dd, J=4.5, 9.1 Hz, 1H), 3.66 (s, 3H),
3.43-3.49 (m, 2H), 2.08-2.14 (m, 1H), 1.82-1.89 (m, 1H). LC-MS:
Anal. Calcd. for C.sub.7H.sub.13NO.sub.5: 191; found: 192
(M+H).sup.+.
Cap-88
##STR00101##
[0315] A mixture of L-valine (1.0 g, 8.54 mmol), 3-bromopyridine
(1.8 mL, 18.7 mmol), K.sub.2CO.sub.3 (2.45 g, 17.7 mmol) and CuI
(169 mg, 0.887 mmol) in DMSO (10 mL) was heated at 100.degree. C.
for 12 h. The reaction mixture was cooled to rt, poured into
H.sub.2O (ca. 150 mL) and washed with EtOAc (.times.2). The organic
layers were extracted with a small amount of H.sub.2O and the
combined aq phases were acidified to ca. pH 2 with 6N HCl. The
volume was reduced to about one-third and 20 g of cation exchange
resin (Strata) was added. The slurry was allowed to stand for 20
min and loaded onto a pad of cation exchange resin (Strata) (ca. 25
g). The pad was washed with H.sub.2O (200 mL), MeOH (200 mL), and
then NH.sub.3 (3M in MeOH, 2.times.200 mL). The appropriate
fractions was concentrated in vacuo and the residue (ca. 1.1 g) was
dissolved in H.sub.2O, frozen and lyophyllized. The title compound
was obtained as a foam (1.02 g, 62%). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.00 (s, br, 1H), 7.68-7.71 (m, 1H), 7.01 (s,
br, 1H), 6.88 (d, J=7.5 Hz, 1H), 5.75 (s, br, 1H), 3.54 (s, 1H),
2.04-2.06 (m, 1H), 0.95 (d, J=6.0 Hz, 3H), 0.91 (d, J=6.6 Hz, 3H).
LC-MS: Anal. Calcd. for C.sub.10H.sub.14N.sub.2O.sub.2: 194; found:
195 (M+H).sup.+.
Cap-89
##STR00102##
[0317] A mixture of L-valine (1.0 g, 8.54 mmol), 5-bromopyrimidine
(4.03 g, 17.0 mmol), K.sub.2CO.sub.3 (2.40 g, 17.4 mmol) and CuI
(179 mg, 0.94 mmol) in DMSO (10 mL) was heated at 100.degree. C.
for 12 h. The reaction mixture was cooled to RT, poured into
H.sub.2O (ca. 150 mL) and washed with EtOAc (.times.2). The organic
layers were extracted with a small amount of H.sub.2O and the
combined aq phases were acidified to ca. pH 2 with 6N HCl. The
volume was reduced to about one-third and 20 g of cation exchange
resin (Strata) was added. The slurry was allowed to stand for 20
min and loaded onto a pad of cation exchange resin (Strata) (ca. 25
g). The pad was washed with H.sub.2O (200 mL), MeOH (200 mL), and
then NH.sub.3 (3M in MeOH, 2.times.200 mL). The appropriate
fractions was concentrated in vacuo and the residue (ca. 1.1 g) was
dissolved in H.sub.2O, frozen and lyophyllized. The title compound
was obtained as a foam (1.02 g, 62%). .sup.1H NMR (400 MHz,
CD.sub.3OD) showed the mixture to contain valine and the purity
could not be estimated. The material was used as is in subsequent
reactions. LC-MS: Anal. Calcd. for C.sub.9H.sub.13N.sub.3O.sub.2:
195; found: 196 (M+H).sup.+.
Cap-90
##STR00103##
[0319] Cap-90 was prepared according to the method described for
the preparation of Cap-1. The crude material was used as is in
subsequent steps. LC-MS: Anal. Calcd. for C.sub.11H.sub.15NO.sub.2:
193; found: 192 (M-H).sup.-.
Cap-91 to Cap-116
[0320] The following Caps were prepared according to the method
used for preparation of Cap-51 unless noted otherwise:
TABLE-US-00008 Cap Structure LC-MS Cap-91 ##STR00104## LC-MS: Anal.
Calcd. for C.sub.11H.sub.13NO.sub.4: 223; found: 222 (M - H).sup.-.
Cap-92 ##STR00105## LC-MS: Anal. Calcd. for
C.sub.11H.sub.13NO.sub.4: 223; found: 222 (M - H).sup.-. Cap-93
##STR00106## LC-MS: Anal. Calcd. for
C.sub.10H.sub.12N.sub.2O.sub.4: 224; found: 225 (M + H).sup.+.
Cap-94 ##STR00107## LC-MS: Anal. Calcd. for
C.sub.8H.sub.11N.sub.3O.sub.4: 213; found: 214 (M + H).sup.+.
Cap-95 ##STR00108## LC-MS: Anal. Calcd. for
C.sub.13H.sub.17NO.sub.4: 251; found: 250 (M - H).sup.-. Cap-96
##STR00109## LC-MS: Anal. Calcd. for C.sub.12H.sub.15NO.sub.4: 237;
found: 236 (M - H).sup.-. Cap-97 ##STR00110## LC-MS: Anal. Calcd.
for C.sub.9H.sub.15NO.sub.4: 201; found: 200 (M - H).sup.-. Cap-98
##STR00111## LC-MS: Anal. Calcd. for C.sub.9H.sub.15NO.sub.4: 201;
found: 202 (M + H).sup.+. Cap-99 ##STR00112## .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 3.88- 3.94 (m, 1H), 3.60, 3.61 (s, 3H), 2.80
(m, 1H), 2.20 (m 1H), 1.82-1.94 (m, 3H), 1.45-1.71 (m, 2H). Cap-99a
##STR00113## .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 3.88- 3.94
(m, 1H), 3.60, 3.61 (s, 3H), 2.80 (m, 1H), 2.20 (m 1H), 1.82-1.94
(m, 3H), 1.45-1.71 (m, 2H). Cap-100 ##STR00114## LC-MS: Anal.
Calcd. for C.sub.12H.sub.14NO.sub.4F: 255; found: 256 (M +
H).sup.+. Cap-101 ##STR00115## LC-MS: Anal. Calcd. for
C.sub.11H.sub.13NO.sub.4: 223; found: 222 (M - H).sup.-. Cap-102
##STR00116## LC-MS: Anal. Calcd. for C.sub.11H.sub.13NO.sub.4: 223;
found: 222 (M - H).sup.- Cap-103 ##STR00117## LC-MS: Anal. Calcd.
for C.sub.10H.sub.12N.sub.2O.sub.4: 224; found: 225 (M + H).sup.+.
Cap-104 ##STR00118## .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 3.60
(s, 3H), 3.50-3.53 (m, 1H), 2.66-2.69 and 2.44-2.49 (m, 1H),
1.91-2.01 (m, 2H), 1.62-1.74 (m, 4H), 1.51-1.62 (m, 2H). Cap-105
##STR00119## .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 3.60 (s,
3H), 3.33-3.35 (m, 1H, partially obscured by solvent), 2.37-2.41
and 2.16-2.23 (m, 1H), 1.94-2.01 (m, 4H), 1.43-1.53 (m, 2H),
1.17-1.29 (m, 2H). Cap-106 ##STR00120## Prepared from cis-4-
aminocyclohexane carboxylic acid and acetaldehyde by employing a
similar procedure described for the synthesis of Cap-2. The crude
HCl salt was passed through MCX (MeOH/H.sub.2O/CH.sub.2Cl.sub.2
wash; 2 N NH.sub.3/MeOH elution) to afford an oil, which was
dissolved in CH.sub.3CN/H.sub.2O and lyophilized to afford a tan
solid. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 3.16 (q, J = 7.3
Hz, 4H), 2.38-2.41 (m, 1H), 2.28-2.31 (m, 2H), 1.79-1.89 (m, 2H),
1.74 (app, ddd J = 3.5, 12.5, 15.9 Hz, 2H), 1.46 (app dt J = 4.0,
12.9 Hz, 2H), 1.26 (t, J = 7.3 Hz, 6H) Cap-107 ##STR00121## LC-MS:
Anal. Calcd. for C.sub.8H.sub.10N.sub.2O.sub.4S: 230; found: 231 (M
+ H).sup.+. Cap-108 ##STR00122## LC-MS: Anal. Calcd. for
C.sub.15H.sub.17N.sub.3O.sub.4: 303; found: 304 (M + H).sup.+.
Cap-109 ##STR00123## LC-MS: Anal. Calcd. for
C.sub.10H.sub.12N.sub.2O.sub.4: 224; found: 225 (M + H).sup.+.
Cap-110 ##STR00124## LC-MS: Anal. Calcd. for
C.sub.10H.sub.12N.sub.2O.sub.4: 224; found: 225 (M + H).sup.+.
Cap-111 ##STR00125## LC-MS: Anal. Calcd. for
C.sub.12H.sub.16NO.sub.8P: 333; found: 334 (M + H).sup.+. Cap-112
##STR00126## LC-MS: Anal. Calcd. for
C.sub.13H.sub.14N.sub.2O.sub.4: 262; found: 263 (M + H).sup.+.
Cap-113 ##STR00127## LC-MS: Anal. Calcd. for
C.sub.18H.sub.19NO.sub.5: 329; found: 330 (M + H).sup.+. Cap-114
##STR00128## .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.82- 4.84
(m, 1H), 4.00-4.05 (m, 2H), 3.77 (s, 3H), 2.56 (s, br, 2H) Cap-115
##STR00129## .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.13 (s, br,
1H), 4.13 (s, br, 1H), 3.69 (s, 3H), 2.61 (d, J = 5.0 Hz, 2H), 1.28
(d, J = 9.1 Hz, 3H). Cap-116 ##STR00130## .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 5.10 (d, J = 8.6 Hz, 1H), 3.74-3.83 (m, 1H),
3.69 (s, 3H), 2.54-2.61 (m, 2H), 1.88 (sept, J = 7.0 Hz, 1H), 0.95
(d, J = 7.0 Hz, 6H).
Cap-117 to Cap-123
[0321] For the preparation of Cap-117 to Cap-123 the Boc amino
acids were obtained from commercially sources and were deprotected
by treatment with 25% TFA in CH.sub.2Cl.sub.2. After complete
reaction as judged by LC-MS the solvents were removed in vacuo and
the corresponding TFA salt of the amino acid was carbamoylated with
methyl chloroformate according to the procedure described for
Cap-51.
TABLE-US-00009 Cap Structure LC-MS Cap-117 ##STR00131## LC-MS:
Anal. Calcd. for C.sub.12H.sub.15NO.sub.4: 237; found: 238 (M +
H).sup.+. Cap-118 ##STR00132## LC-MS: Anal. Calcd. for
C.sub.10H.sub.13NO.sub.4S: 243; found: 244 (M + H).sup.+. Cap-119
##STR00133## LC-MS: Anal. Calcd. for C.sub.10H.sub.13NO.sub.4S:
243; found: 244 (M + H).sup.+. Cap-120 ##STR00134## LC-MS: Anal.
Calcd. for C.sub.10H.sub.13NO.sub.4S: 243; found: 244 (M +
H).sup.+. Cap-121 ##STR00135## .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 4.06-4.16 (m, 1H), 3.63 (s, 3H), 3.43 (s, 1H), 2.82 and
2.66 (s, br, 1H), 1.86-2.10 (m, 3H), 1.64-1.76 (m, 2H), 1.44-1.53
(m, 1H). Cap-122 ##STR00136## .sup.1H NMR profile is similar to
that of its stereoisomer, Cap-121. Cap-123 ##STR00137## LC-MS:
Anal. Calcd. for C.sub.27H.sub.26N.sub.2O.sub.6: 474; found: 475 (M
+ H).sup.+.
Cap-124
##STR00138##
[0323] The hydrochloride salt of L-threonine tert-butyl ester was
carbamoylated according to the procedure for Cap-51. The crude
reaction mixture was acidified with 1N HCl to pH-1 and the mixture
was extracted with EtOAc (2.times.50 mL). The combined organic
phases were concentrated in vacuo to give a colorless oil which
solidified on standing. The aqueous layer was concentrated in vacuo
and the resulting mixture of product and inorganic salts was
triturated with EtOAc--CH.sub.2Cl.sub.2-MeOH (1:1:0.1) and then the
organic phase concentrated in vacuo to give a colorless oil which
was shown by LC-MS to be the desired product. Both crops were
combined to give 0.52 g of a solid. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 4.60 (m, 1H), 4.04 (d, J=5.0 Hz, 1H), 1.49 (d,
J=6.3 Hz, 3H). LC-MS: Anal. Calcd. for C.sub.5H.sub.7NO.sub.4: 145;
found: 146 (M+H).sup.+.
Cap-125
##STR00139##
[0325] To a suspension of Pd(OH).sub.2, (20%, 100 mg), aqueous
formaldehyde (37% wt, 4 ml), acetic acid, (0.5 mL) in methanol (15
mL) was added (S)-4-amino-2-(tert-butoxycarbonylamino)butanoic acid
(1 g, 4.48 mmol). The reaction was purged several times with
hydrogen and was stirred overnight with an hydrogen balloon room
temperature. The reaction mixture was filtered through a pad of
diatomaceous earth (CELITE.RTM.), and the volatile component was
removed in vacuo. The resulting crude material was used as is for
the next step. LC-MS: Anal. Calcd. for
C.sub.11H.sub.22N.sub.2O.sub.4: 246; found: 247 (M+H).sup.+.
Cap-126
##STR00140##
[0327] This procedure is a modification of that used to prepare
Cap-51. To a suspension of 3-methyl-L-histidine (0.80 g, 4.70 mmol)
in THF (10 mL) and H.sub.2O (10 mL) at 0.degree. C. was added
NaHCO.sub.3 (0.88 g, 10.5 mmol). The resulting mixture was treated
with ClCO.sub.2Me (0.40 mL, 5.20 mmol) and the mixture allowed to
stir at 0.degree. C. After stirring for ca. 2 h LC-MS showed no
starting material remaining. The reaction was acidified to pH 2
with 6 N HCl.
[0328] The solvents were removed in vacuo and the residue was
suspended in 20 mL of 20% MeOH in CH.sub.2Cl.sub.2. The mixture was
filtered and concentrated to give a light yellow foam (1.21 g,).
LC-MS and .sup.1H NMR showed the material to be a 9:1 mixture of
the methyl ester and the desired product. This material was taken
up in THF (10 mL) and H.sub.2O (10 mL), cooled to 0.degree. C. and
LiOH (249.1 mg, 10.4 mmol) was added. After stirring ca. 1 h LC-MS
showed no ester remaining. Therefore the mixture was acidified with
6N HCl and the solvents removed in vacuo. LC-MS and .sup.1H NMR
confirm the absence of the ester. The title compound was obtained
as its HCl salt contaminated with inorganic salts (1.91 g,
>100%). The compound was used as is in subsequent steps without
further purification. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
8.84, (s, 1H), 7.35 (s, 1H), 4.52 (dd, J=5.0, 9.1 Hz, 1H), 3.89 (s,
3H), 3.62 (s, 3H), 3.35 (dd, J=4.5, 15.6 Hz, 1H, partially obscured
by solvent), 3.12 (dd, J=9.0, 15.6 Hz, 1H). LC-MS: Anal. Calcd. for
C.sub.9H.sub.13N.sub.3O.sub.4: 227.09; found: 228.09
(M+H).sup.+.
Cap-127
##STR00141##
[0330] Cap-127 was prepared according to the method for Cap-126
above starting from
(S)-2-amino-3-(1-methyl-1H-imidazol-4-yl)propanoic acid (1.11 g,
6.56 mmol), NaHCO.sub.3 (1.21 g, 14.4 mmol) and ClCO.sub.2Me (0.56
mL, 7.28 mmol). The title compound was obtained as its HCl salt
(1.79 g, >100%) contaminated with inorganic salts. LC-MS and
.sup.1H NMR showed the presence of ca. 5% of the methyl ester. The
crude mixture was used as is without further purification. .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.90 (s, 1H), 7.35 (s, 1H), 4.48
(dd, J=5.0, 8.6 Hz, 1H), 3.89 (s, 3H), 3.62 (s, 3H), 3.35 (m, 1H),
3.08 (m, 1H); LC-MS: Anal. Calcd. for
C.sub.9H.sub.13N.sub.3O.sub.4: 227.09; found: 228 (M+H).sup.+.
Preparation of Cap-128
##STR00142##
[0332] Step 1. Preparation of (S)-benzyl
2-(tert-butoxycarbonylamino)pent-4-ynoate (cj-27b).
##STR00143##
[0333] To a solution of cj-27a (1.01 g, 4.74 mmol), DMAP (58 mg,
0.475 mmol) and iPr.sub.2NEt (1.7 mL, 9.8 mmol) in CH.sub.2Cl.sub.2
(100 mL) at 0.degree. C. was added Cbz-Cl (0.68 mL, 4.83 mmol). The
solution was allowed to stir for 4 h at 0.degree. C., washed (1N
KHSO.sub.4, brine), dried (Na.sub.2SO.sub.4), filtered, and
concentrated in vacuo. The residue was purified by flash column
chromatography (TLC 6:1 hex:EtOAc) to give the title compound (1.30
g, 91%) as a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.35 (s, 5H), 5.35 (d, br, J=8.1 Hz, 1H), 5.23 (d, J=12.2
Hz, 1H), 5.17 (d, J=12.2 Hz, 1H), 4.48-4.53 (m, 1H), 2.68-2.81 (m,
2H), 2.00 (t, J=2.5 Hz, 1H), 1.44 (s, 9H). LC-MS: Anal. Calcd. for
C.sub.17H.sub.21Na.sub.4: 303; found: 304 (M+H).sup.+.
Step 2. Preparation of (S)-benzyl
3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(tert-butoxycarbonylamino)propanoate
(cj-28)
##STR00144##
[0335] To a mixture of (S)-benzyl
2-(tert-butoxycarbonylamino)pent-4-ynoate (0.50 g, 1.65 mmol),
sodium ascorbate (0.036 g, 0.18 mmol), CuSO.sub.4-5H.sub.2O (0.022
g, 0.09 mmol) and NaN.sub.3 (0.13 g, 2.1 mmol) in DMF-H.sub.2O (5
mL, 4:1) at rt was added BnBr (0.24 mL, 2.02 mmol) and the mixture
was warmed to 65.degree. C. After 5 h LC-MS indicated low
conversion. A further portion of NaN.sub.3 (100 mg) was added and
heating was continued for 12 h. The reaction was poured into EtOAc
and H.sub.2O and shaken. The layers were separated and the aqueous
layer extracted 3.times. with EtOAc and the combined organic phases
washed (H.sub.2O.times.3, brine), dried (Na.sub.2SO.sub.4),
filtered, and concentrated. The residue was purified by flash
(BIOTAGE.RTM., 40+M 0-5% MeOH in CH.sub.2Cl.sub.2; TLC 3% MeOH in
CH.sub.2Cl.sub.2) to afford a light yellow oil which solidified on
standing (748.3 mg, 104%). The NMR was consistent with the desired
product but suggests the presence of DMF. The material was used as
is without further purification. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.84 (s, 1H), 7.27-7.32 (m, 10H), 5.54 (s,
2H), 5.07 (s, 2H), 4.25 (m, 1H), 3.16 (dd, J=1.0, 5.3 Hz, 1H), 3.06
(dd, J=5.3, 14.7 Hz), 2.96 (dd, J=9.1, 14.7 Hz, 1H), 1.31 (s, 9H).
LC-MS: Anal. Calcd. for C.sub.24H.sub.28N.sub.4O.sub.4: 436; found:
437 (M+H).sup.+.
Step 3. Preparation of (S)-benzyl
3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(methoxycarbonylamino)propanoate
(cj-29)
##STR00145##
[0337] A solution of (S)-benzyl
3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(tert-butoxycarbonylamino)propanoate
(0.52 g, 1.15 mmol) in CH.sub.2Cl.sub.2 was added TFA (4 mL). The
mixture was allowed to stir at room temperature for 2 h. The
mixture was concentrated in vacuo to give a colorless oil which
solidified on standing. This material was dissolved in
THF--H.sub.2O and cooled to 0.degree. C. Solid NaHCO.sub.3 (0.25 g,
3.00 mmol) was added followed by ClCO.sub.2Me (0.25 mL, 3.25 mmol).
After stirring for 1.5 h the mixture was acidified to pH.about.2
with 6N HCl and then poured into H.sub.2O-EtOAc. The layers were
separated and the aq phase extracted 2.times. with EtOAc. The
combined org layers were washed (H.sub.2O, brine), dried
(Na.sub.2SO.sub.4), filtered, and concentrated in vacuo to give a
colorless oil (505.8 mg, 111%, NMR suggested the presence of an
unidentified impurity) which solidified while standing on the pump.
The material was used as is without further purification. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 7.87 (s, 1H), 7.70 (d, J=8.1
Hz, 1H), 7.27-7.32 (m, 10H), 5.54 (s, 2H), 5.10 (d, J=12.7 Hz, 1H),
5.06 (d, J=12.7 Hz, 1H), 4.32-4.37 (m, 1H), 3.49 (s, 3H), 3.09 (dd,
J=5.6, 14.7 Hz, 1H), 2.98 (dd, J=9.6, 14.7 Hz, 1H). LC-MS: Anal.
Calcd. for C.sub.21H.sub.22N.sub.4O.sub.4: 394; found: 395
(M+H).sup.+.
Step 4. Preparation of
(S)-2-(methoxycarbonylamino)-3-(1H-1,2,3-triazol-4-yl)propanoic
acid (Cap-128)
##STR00146##
[0339] (S)-Benzyl
3-(1-benzyl-1H-1,2,3-triazol-4-yl)-2-(methoxycarbonylamino)propanoate
(502 mg, 1.11 mmol) was hydrogenated in the presence of Pd--C (82
mg) in MeOH (5 mL) at atmospheric pressure for 12 h. The mixture
was filtered through diatomaceous earth (CELITE.RTM.) and
concentrated in vacuo.
(S)-2-(methoxycarbonylamino)-3-(1H-1,2,3-triazol-4-yl)propanoic
acid was obtained as a colorless gum (266 mg, 111%) which was
contaminated with ca. 10% of the methyl ester. The material was
used as is without further purification. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 12.78 (s, br, 1H), 7.59 (s, 1H), 7.50 (d,
J=8.0 Hz, 1H), 4.19-4.24 (m, 1H), 3.49 (s, 3H), 3.12 (dd, J=4.8 Hz,
14.9 Hz, 1H), 2.96 (dd, J=9.9, 15.0 Hz, 1H). LC-MS: Anal. Calcd.
for C.sub.7H.sub.10N.sub.4O.sub.4: 214; found: 215 (M+H).sup.+.
Preparation of Cap-129
##STR00147##
[0340] Step 1. Preparation of
(S)-2-(benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid
(cj-31)
##STR00148##
[0342] A suspension of (S)-benzyl 2-oxooxetan-3-ylcarbamate (0.67
g, 3.03 mmol), and pyrazole (0.22 g, 3.29 mmol) in CH.sub.3CN (12
mL) was heated at 50.degree. C. for 24 h. The mixture was cooled to
rt overnight and the solid filtered to afford
(S)-2-(benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid
(330.1 mg). The filtrate was concentrated in vacuo and then
triturated with a small amount of CH.sub.3CN (ca. 4 mL) to afford a
second crop (43.5 mg). Total yield 370.4 mg (44%). m.p.
165.5-168.degree. C. lit m.p. 168.5-169.5 [Vederas et al., J. Am.
Chem. Soc., 107:7105 (1985)]. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.51 (d, J=2.0, 1H), 7.48 (s, J=1.5 Hz, 1H), 7.24-7.34 (m,
5H), 6.23 m, 1H), 5.05 (d, 12.7H, 1H), 5.03 (d, J=12.7 Hz, 1H),
4.59-4.66 (m, 2H), 4.42-4.49 (m, 1H). LC-MS: Anal. Calcd. for
C.sub.14H.sub.15N.sub.3O.sub.4: 289; found: 290 (M+H).sup.+.
Step 2. Preparation of
(S)-2-(methoxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid
(Cap-129)
##STR00149##
[0344] (S)-2-(Benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic
acid (0.20 g, 0.70 mmol) was hydrogenated in the presence of Pd--C
(45 mg) in MeOH (5 mL) at atmospheric pressure for 2 h. The product
appeared to be insoluble in MeOH, therefore the reaction mixture
was diluted with 5 mL H.sub.2O and a few drops of 6N HCl. The
homogeneous solution was filtered through diatomaceous earth
(CELITE.RTM.), and the MeOH removed in vacuo. The remaining
solution was frozen and lyophyllized to give a yellow foam (188.9
mg). This material was suspended in THF--H.sub.2O (1:1, 10 mL) and
then cooled to 0.degree. C. To the cold mixture was added
NaHCO.sub.3 (146.0 mg, 1.74 mmol) carefully (evolution of
CO.sub.2). After gas evolution had ceased (ca. 15 min) ClCO.sub.2Me
(0.06 mL, 0.78 mmol) was added dropwise. The mixture was allowed to
stir for 2 h and was acidified to pH.about.2 with 6N HCl and poured
into EtOAc. The layers were separated and the aqueous phase
extracted with EtOAC (.times.5). The combined organic layers were
washed (brine), dried (Na.sub.2SO.sub.4), filtered, and
concentrated to give the title compound as a colorless solid (117.8
mg, 79%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 13.04 (s,
1H), 7.63 (d, J=2.6 Hz, 1H), 7.48 (d, J=8.1 Hz, 1H), 7.44 (d, J=1.5
Hz, 1H), 6.19 (app t, J=2.0 Hz, 1H), 4.47 (dd, J=3.0, 12.9 Hz, 1H),
4.29-4.41 (m, 2H), 3.48 (s, 3H). LC-MS: Anal. Calcd. for
C.sub.8H.sub.11N.sub.3O.sub.4: 213; found: 214 (M+H).sup.+.
Cap-130
##STR00150##
[0346] Cap-130 was prepared by acylation of commercially available
(R)-phenylglycine analogous to the procedure given in: Calmes, M.
et al., Tetrahedron, 43(10):2285 (1987).
Cap-131
##STR00151##
[0348] Step a: Dimethylcarbamoyl chloride (0.92 mL, 10 mmol) was
added slowly to a solution of (S)-benzyl 2-amino-3-methylbutanoate
hydrochloride (2.44 g; 10 mmol) and Hunig's base (3.67 mL, 21 mmol)
in THF (50 mL). The resulting white suspension was stirred at room
temperature overnight (16 hours) and concentrated under reduced
pressure. The residue was partitioned between ethyl acetate and
water. The organic layer was washed with brine, dried (MgSO.sub.4),
filtered, and concentrated under reduced pressure. The resulting
yellow oil was purified by flash chromatography, eluting with ethyl
acetate:hexanes (1:1). Collected fractions were concentrated under
vacuum providing 2.35 g (85%) of clear oil. .sup.1HNMR (300 MHz,
DMSO-d.sub.6) .delta. ppm 0.84 (d, J=6.95 Hz, 3H), 0.89 (d, J=6.59
Hz, 3H), 1.98-2.15 (m, 1H), 2.80 (s, 6H), 5.01-5.09 (m, J=12.44 Hz,
1H), 5.13 (d, J=12.44 Hz, 1H), 6.22 (d, J=8.05 Hz, 1H), 7.26-7.42
(m, 5H). LC (Cond. 1): RT=1.76 min; MS: Anal. Calcd. for
[M+H].sup.+ C.sub.16H.sub.22N.sub.2O.sub.3: 279.17; found
279.03.
[0349] Step b: To an MeOH (50 mL) solution of the intermediate
prepared above (2.35 g; 8.45 mmol) was added Pd/C (10%; 200 mg) and
the resulting black suspension was flushed with N.sub.2 (3.times.)
and placed under 1 atm of H.sub.2. The mixture was stirred at room
temperature overnight and filtered though a microfiber filter to
remove the catalyst. The resulting clear solution was then
concentrated under reduced pressure to obtain 1.43 g (89%) of
Cap-131 as a white foam, which was used without further
purification. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 0.87
(d, J=4.27 Hz, 3H), 0.88 (d, J=3.97 Hz, 3H), 1.93-2.11 (m, 1H),
2.80 (s, 6H), 3.90 (dd, J=8.39, 6.87 Hz, 1H), 5.93 (d, J=8.54 Hz,
1H), 12.36 (s, 1H). LC (Cond. 1): RT=0.33 min; MS: Anal. Calcd. for
[M+H].sup.+ C.sub.8H.sub.17N.sub.2O.sub.3: 189.12; found
189.04.
Cap-132
##STR00152##
[0351] Cap-132 was prepared from (S)-benzyl 2-aminopropanoate
hydrochloride according to the method described for Cap-131.
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 1.27 (d, J=7.32 Hz,
3H), 2.80 (s, 6H), 4.06 (qt, 1H), 6.36 (d, J=7.32 Hz, 1H), 12.27
(s, 1H). LC (Cond. 1): RT=0.15 min; MS: Anal. Calcd. for
[M+H].sup.+ C.sub.6H.sub.13N.sub.2O.sub.3: 161.09; found
161.00.
Cap-133
##STR00153##
[0353] Cap-133 was prepared from (S)-tert-butyl
2-amino-3-methylbutanoate hydrochloride and 2-fluoroethyl
chloroformate according to the method described for Cap-47. .sup.1H
NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 0.87 (t, J=6.71 Hz, 6H),
1.97-2.10 (m, 1H), 3.83 (dd, J=8.39, 5.95 Hz, 1H), 4.14-4.18 (m,
1H), 4.20-4.25 (m, 1H), 4.50-4.54 (m, 1H), 4.59-4.65 (m, 1H), 7.51
(d, J=8.54 Hz, 1H), 12.54 (s, 1H).
Cap-134
##STR00154##
[0355] Cap-134 was prepared from (S)-diethyl alanine and methyl
chloroformate according to the method described for Cap-51. .sup.1H
NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 0.72-0.89 (m, 6H),
1.15-1.38 (m, 4H), 1.54-1.66 (m, 1H), 3.46-3.63 (m, 3H), 4.09 (dd,
J=8.85, 5.19 Hz, 1H), 7.24 (d, J=8.85 Hz, 1H), 12.55 (s, 1H). LC
(Cond. 2): RT=0.66 min; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.9H.sub.18NO.sub.4: 204.12; found 204.02.
Cap-135
##STR00155##
[0357] A solution of D-2-amino-(4-fluorophenyl)acetic acid (338 mg,
2.00 mmol), 1N HCl in diethylether (2.0 mL, 2.0 mmol) and formalin
(37%, 1 mL) in methanol (5 mL) was subjected to balloon
hydrogenation over 10% palladium on carbon (60 mg) for 16 h at
25.degree. C. The mixture was then filtered through CELITE.RTM. to
afford the HCl salt of Cap-135 as a white foam (316 mg, 80%).
.sup.1H NMR (300 MHz, MeOH-d.sub.4) .delta. 7.59 (dd, J=8.80, 5.10
Hz, 2H), 7.29 (t, J=8.6 Hz, 2H), 5.17 (s, 1H), 3.05 (v br s, 3H),
2.63 (v br s, 3H); R.sub.t=0.19 min (Cond.-MS-W5); 95% homogenity
index; LRMS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.13FNO.sub.2: 198.09; found: 198.10.
Cap-136
##STR00156##
[0359] To a cooled (-50.degree. C.) suspension of
1-benzyl-1H-imidazole (1.58 g, 10.0 mmol) in anhydrous diethyl
ether (50 mL) under nitrogen was added n-butyl lithium (2.5 M in
hexanes, 4.0 mL, 10.0 mmol) dropwise. After being stirred for 20
min at -50.degree. C., dry carbon dioxide (passed through Drierite)
was bubbled into the reaction mixture for 10 min before it was
allowed to warm up to 25.degree. C. The heavy precipitate which
formed on addition of carbon dioxide to the reaction mixture was
filtered to yield a hygroscopic, white solid which was taken up in
water (7 mL), acidified to pH=3, cooled, and induced to crystallize
with scratching. Filtration of this precipitate gave a white solid
which was suspended in methanol, treated with 1N HCl/diethyl ether
(4 mL) and concentrated in vacuo. Lyophilization of the residue
from water (5 mL) afforded the HCl salt of Cap-136 as a white solid
(817 mg, 40%). .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 7.94 (d,
J=1.5 Hz, 1H), 7.71 (d, J=1.5 Hz, 1H), 7.50-7.31 (m, 5H), 5.77 (s,
2H); R.sub.t=0.51 min (Cond.-MS-W5); 95% homogenity index; LRMS:
Anal. Calc. for [M+H].sup.+ C.sub.11H.sub.12N.sub.2O.sub.2: 203.08;
found: 203.11.
Cap-137
##STR00157##
[0360] Cap-137, Step a
##STR00158##
[0362] A suspension of 1-chloro-3-cyanoisoquinoline (188 mg, 1.00
mmol; prepared according to the procedure in WO 2003/099274) (188
mg, 1.00 mmol), cesium fluoride (303.8 mg, 2.00 mmol),
bis(tri-tert-butylphosphine)palladium dichloride (10 mg, 0.02 mmol)
and 2-(tributylstannyl)furan (378 .mu.L, 1.20 mmol) in anhydrous
dioxane (10 mL) under nitrogen was heated at 80.degree. C. for 16 h
before it was cooled to 25.degree. C. and treated with saturated,
aqueous potassium fluoride solution with vigorous stirring for 1 h.
The mixture was partitioned between ethyl acetate and water and the
organic phase was separated, washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. Purification of the
residue on silica gel (elution with 0% to 30% ethyl
acetate/hexanes) afforded Cap-137, step a as a white solid which
was used as is (230 mg, 105%). R.sub.t=1.95 min (Cond.-MS-W2); 90%
homogeneity index; LRMS: Anal. Calc. for [M+H].sup.+
C.sub.14H.sub.8N.sub.2O: 221.07; found: 221.12.
Cap-137
##STR00159##
[0364] To a suspension of Cap-137, step a (110 mg, 0.50 mmol) and
sodium periodate (438 mg, 2.05 mmol) in carbon tetrachloride (1
mL), acetonitrile (1 mL) and water (1.5 mL) was added ruthenium
trichloride hydrate (2 mg, 0.011 mmol). The mixture was stirred at
25.degree. C. for 2 h and then partitioned between dichloromethane
and water. The aqueous layer was separated, extracted twice more
with dichloromethane and the combined dichloromethane extracts were
dried over Na.sub.2SO.sub.4, filtered and concentrated. Trituration
of the residue with hexanes afforded Cap-137 (55 mg, 55%) as a
grayish-colored solid. R.sub.t=1.10 min (Cond.-MS-W2); 90%
homogeneity index; LC-MS: Anal. Calc. for [M+H].sup.+
C.sub.11H.sub.8N.sub.2O.sub.2: 200.08; found: 200.08.
Cap-138 to Cap-158
[0365] Synthetic Strategy. Method A.
##STR00160##
Cap-138
##STR00161##
[0366] Cap-138, Step a
##STR00162##
[0368] To a stirred suspension of 5-hydroxisoquinoline (prepared
according to the procedure in WO 2003/099274) (2.0 g, 13.8 mmol)
and triphenylphosphine (4.3 g, 16.5 mmol) in dry tetrahydrofuran
(20 mL) was added dry methanol (0.8 mL) and diethyl
azodicarboxylate (3.0 mL, 16.5 mmol) portionwise. The mixture was
stirred at room temperature for 20 h before it was diluted with
ethyl acetate and washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was preabsorbed onto silica
gel and purified (elution with 40% ethyl acetate/hexanes) to afford
Cap-138, step a as a light yellow solid (1.00 g, 45%). .sup.1H NMR
(CDCl.sub.3, 500 MHz) .delta. 9.19 (s, 1H), 8.51 (d, J=6.0 Hz, 1H),
7.99 (d, J=6.0 Hz, 1H), 7.52-7.50 (m, 2H), 7.00-6.99 (m, 1H), 4.01
(s, 3H); R.sub.t=0.66 min (Cond. D2); 95% homogeneity index; LC-MS:
Anal. Calc. for [M+H].sup.+ C.sub.10H.sub.10NO: 160.08; found
160.10.
Cap-138, Step b
##STR00163##
[0370] To a stirred solution of Cap-138, step a (2.34 g, 14.7 mmol)
in anhydrous dichloromethane (50 mL) at room temperature was added
meta-chloroperbenzoic acid (77%, 3.42 g, 19.8 mmol) in one portion.
After being stirred for 20 h, powdered potassium carbonate (2.0 g)
was added and the mixture was stirred for 1 h at room temperature
before it was filtered and concentrated to afford Cap-138, step b
as a pale, yellow solid which was sufficiently pure to carry
forward (2.15 g, 83.3%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
8.73 (d, J=1.5 Hz, 1H), 8.11 (dd, J=7.3, 1.7 Hz, 1H), 8.04 (d,
J=7.1 Hz, 1H), 7.52 (t, J=8.1 Hz, 1H), 7.28 (d, J=8.3 Hz, 1H), 6.91
(d, J=7.8 Hz, 1H), 4.00 (s, 3H); R.sub.t=0.92 min, (Cond.-D1); 90%
homogenity index; LC-MS: Anal. Calc. for [M+H].sup.+
C.sub.10H.sub.10NO.sub.2: 176.07; found: 176.0.
Cap-138, Step c
##STR00164##
[0372] To a stirred solution of Cap-138, step b (0.70 g, 4.00 mmol)
and triethylamine (1.1 mL, 8.00 mmol) in dry acetonitrile (20 mL)
at room temperature under nitrogen was added trimethylsilylcyanide
(1.60 mL, 12.00 mmol). The mixture was heated at 75.degree. C. for
20 h before it was cooled to room temperature, diluted with ethyl
acetate and washed with saturated sodium bicarbonate solution and
brine prior to drying over Na.sub.2SO.sub.4 and solvent
concentration. The residue was flash chromatographed on silica gel
(elution with 5% ethyl acetate/hexanes) to 25% ethyl
acetate/hexanes to afford Cap-138, step c (498.7 mg) as a white,
crystalline solid along with 223 mg of additional Cap-138, step c
recovered from the filtrate. .sup.1H NMR (CDCl.sub.3, 500 MHz)
.delta. 8.63 (d, J=5.5 Hz, 1H), 8.26 (d, J=5.5 Hz, 1H), 7.88 (d,
J=8.5 Hz, 1H), 7.69 (t, J=8.0 Hz, 1H), 7.08 (d, J=7.5 Hz, 1H), 4.04
(s, 3H); R.sub.t=1.75 min, (Cond.-D1); 90% homogeneity index;
LC-MS: Anal. Calc. for [M+H].sup.+ C.sub.11H.sub.9N.sub.2O: 185.07;
found: 185.10.
Cap-138
##STR00165##
[0374] Cap-138, step c (0.45 g, 2.44 mmol) was treated with 5N
sodium hydroxide solution (10 mL) and the resulting suspension was
heated at 85.degree. C. for 4 h, cooled to 25.degree. C., diluted
with dichloromethane and acidified with 1N hydrochloric acid. The
organic phase was separated, washed with brine, dried over
Na.sub.2SO.sub.4, concentrated to 1/4 volume and filtered to afford
Cap-138 as a yellow solid (0.44 g, 88.9%). .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 13.6 (br s, 1H), 8.56 (d, J=6.0 Hz,
1H), 8.16 (d, J=6.0 Hz, 1H), 8.06 (d, J=8.8 Hz, 1H), 7.71-7.67 (m,
1H), 7.30 (d, J=8.0 Hz, 1H), 4.02 (s, 3H); R.sub.t=0.70 min
(Cond.-D1); 95% homogenity index; LC-MS: Anal. Calc. for
[M+H].sup.+ C.sub.11H.sub.10NO.sub.3: 204.07; found: 204.05.
Synthetic Strategy. Method B (Derived from Tetrahedron Letters,
42:6707 (2001)).
##STR00166##
Cap-139
##STR00167##
[0375] Cap-139, Step a
##STR00168##
[0377] To a thick-walled, screw-top vial containing an
argon-degassed suspension of 1-chloro-6-methoxyisoquinoline (1.2 g,
6.2 mmol; prepared according to the procedure in WO 2003/099274),
potassium cyanide (0.40 g, 6.2 mmol),
1,5-bis(diphenylphosphino)pentane (0.27 g, 0.62 mmol) and palladium
(II) acetate (70 mg, 0.31 mmol) in anhydrous toluene (6 mL) was
added N,N,N',N'-tetramethylethylenediamine (0.29 mL, 2.48 mmol).
The vial was sealed, heated at 150.degree. C. for 22 h and then
allowed to cool to 25.degree. C. The reaction mixture was diluted
with ethyl acetate, washed with water and brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified on silica gel eluting with 5% ethyl acetate/hexanes to 25%
ethyl acetate/hexanes to afford Cap-139, step a as a white solid
(669.7 mg). .sup.1H NMR (CDCl.sub.3, 500 MHz) .delta. 8.54 (d,
J=6.0 Hz, 1H), 8.22 (d, J=9.0 Hz, 1H), 7.76 (d, J=5.5 Hz, 1H),
7.41-7.39 (m, 1H), 7.13 (d, J=2.0 Hz, 1H), 3.98 (s, 3H);
R.sub.t=1.66 min (Cond.-D1); 90% homogenity index; LC-MS: Anal.
Calc. for [M+H].sup.+ C.sub.11H.sub.9N.sub.2O: 185.07; found:
185.20.
Cap-139
##STR00169##
[0379] Cap-139 was prepared from the basic hydrolysis of Cap-139,
step a with 5N NaOH according to the procedure described for
Cap-138. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 13.63 (v br s,
1H), 8.60 (d, J=9.3 Hz, 1H), 8.45 (d, J=5.6 Hz, 1H), 7.95 (d, J=5.9
Hz, 1H), 7.49 (d, J=2.2 Hz, 1H), 7.44 (dd, J=9.3, 2.5 Hz, 1H), 3.95
(s, 3H); R.sub.t=0.64 min (Cond.-D1); 90% homogenity index; LC-MS:
Anal. Calc. for [M+H].sup.+ C.sub.11H.sub.10NO.sub.3: 204.07;
found: 204.05.
Cap-140
##STR00170##
[0380] Cap-140, Step a
##STR00171##
[0382] To a vigorously-stirred mixture of
1,3-dichloro-5-ethoxyisoquinoline (482 mg, 2.00 mmol; prepared
according to the procedure in WO 2005/051410), palladium (II)
acetate (9 mg, 0.04 mmol), sodium carbonate (223 mg, 2.10 mmol) and
1,5-bis(diphenylphosphino)pentane (35 mg, 0.08 mmol) in dry
dimethylacetamide (2 mL) at 25.degree. C. under nitrogen was added
N,N,N',N'-tetramethylethylenediamine (60 mL, 0.40 mmol). After 10
min, the mixture was heated to 150.degree. C., and then a stock
solution of acetone cyanohydrin (prepared from 457 .mu.L of acetone
cyanohydrin in 4.34 mL DMA) was added in 1 mL portions over 18 h
using a syringe pump. The mixture was then partitioned between
ethyl acetate and water and the organic layer was separated, washed
with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated.
The residue was purified on silica gel eluting with 10% ethyl
acetate/hexanes to 40% ethyl acetate/hexanes to afford Cap-140,
step a as a yellow solid (160 mg, 34%). R.sub.t=2.46 min
(Cond.-MS-W2); 90% homogenity index; LC-MS: Anal. Calc. for
[M+H].sup.+ C.sub.12H.sub.9ClN.sub.2O: 233.05; found: 233.08.
Cap-140
##STR00172##
[0384] Cap-140 was prepared by the acid hydrolysis of Cap-140, step
a with 12N HCl as described in the procedure for the preparation of
Cap-141, described below. R.sub.t=2.24 min (Cond.-MS-W2); 90%
homogenity index; LC-MS: Anal. Calc. for [M+H].sup.+
C.sub.12H.sub.11ClNO.sub.3: 252.04; found: 252.02.
Cap-141
##STR00173##
[0385] Cap-141, Step a
##STR00174##
[0387] Cap-141, step a was prepared from
1-bromo-3-fluoroisoquinoline (prepared from
3-amino-1-bromoisoquinoline using the procedure outlined in J. Med.
Chem., 13:613 (1970)) as described in the procedure for the
preparation of Cap-140, step a (vide supra). .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.35 (d, J=8.5 Hz, 1H), 7.93 (d, J=8.5 Hz, 1H),
7.83 (t, J=7.63 Hz, 1H), 7.77-7.73 (m, 1H), 7.55 (s, 1H);
R.sub.t=1.60 min (Cond.-D1); 90% homogenity index; LC-MS: Anal.
Calc. for [M+H].sup.+ C.sub.10H.sub.6FN.sub.2: 173.05; found:
172.99.
Cap-141
##STR00175##
[0389] Cap-141, step a (83 mg, 0.48 mmol) was treated with 12N HCl
(3 mL) and the resulting slurry was heated at 80.degree. C. for 16
h before it was cooled to room temperature and diluted with water
(3 mL). The mixture was stirred for 10 min and then filtered to
afford Cap-141 as an off-white solid (44.1 mg, 47.8%). The filtrate
was diluted with dichloromethane and washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated to afford additional Cap-141
which was sufficiently pure to be carried forward directly (29.30
mg, 31.8%). .sup.1H NMR (DMSO-d.sub.6, 500 MHz) .delta. 14.0 (br s,
1H), 8.59-8.57 (m, 1H), 8.10 (d, J=8.5 Hz, 1H), 7.88-7.85 (m, 2H),
7.74-7.71 (m, 1H); R.sub.t=1.33 min (Cond.-D1); 90% homogenity
index; LC-MS: Anal. Calc. for [M+H].sup.+ C.sub.10H.sub.7FNO.sub.2:
192.05; found: 191.97.
Cap-142
##STR00176##
[0390] Cap-142, Step a
##STR00177##
[0392] Cap-142, step a was prepared from 4-bromoisoquinoline
N-oxide as described in the two-step procedure for the preparation
of Cap-138, steps b and c. R.sub.t=1.45 min (Cond.-MS-W1); 90%
homogenity index; LC-MS: Anal. Calc. for [M+H].sup.+
C.sub.10H.sub.6BrN.sub.2: 232.97; found: 233.00.
Cap-142, Step b
##STR00178##
[0394] To an argon-degassed suspension of Cap-142, step a (116 mg,
0.50 mmol), potassium phosphate tribasic (170 mg, 0.80 mmol),
palladium (II) acetate (3.4 mg, 0.015 mmol) and
2-(dicyclohexylphosphino)biphenyl (11 mg, 0.03 mmol) in anhydrous
toluene (1 mL) was added morpholine (61 .mu.L, 0.70 mmol). The
mixture was heated at 100.degree. C. for 16 h, cooled to 25.degree.
C. and filtered through diatomaceous earth (CELITE.RTM.).
Purification of the residue on silica gel, eluting with 10% to 70%
ethyl acetate/hexanes afforded Cap-142, step b (38 mg, 32%) as a
yellow solid, which was carried forward directly. R.sub.t=1.26 min
(Cond.-MS-W1); 90% homogenity index; LC-MS: Anal. Calc. for
[M+H].sup.+ C.sub.14H.sub.14N.sub.3O: 240.11; found: 240.13.
Cap-142
##STR00179##
[0396] Cap-142 was prepared from Cap-142, step b with 5N sodium
hydroxide as described in the procedure for Cap-138. R.sub.t=0.72
min (Cond.-MS-W1); 90% homogenity index; LC-MS: Anal. Calc. for
[M+H].sup.+ C.sub.14H.sub.15N.sub.2O.sub.3: 259.11; found:
259.08.
Cap-143
##STR00180##
[0397] Cap-143, Step a
##STR00181##
[0399] To a stirred solution of 3-amino-1-bromoisoquinoline (444
mg, 2.00 mmol) in anhydrous dimethylformamide (10 mL) was added
sodium hydride (60%, unwashed, 96 mg, 2.4 mmol) in one portion. The
mixture was stirred at 25.degree. C. for 5 min before 2-bromoethyl
ether (90%, 250 .mu.L, 2.00 mmol) was added. The mixture was
stirred further at 25.degree. C. for 5 h and at 75.degree. C. for
72 h before it was cooled to 25.degree. C., quenched with saturated
ammonium chloride solution and diluted with ethyl acetate. The
organic layer was separated, washed with water and brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated. Purification of
the residue on silica gel eluting with 0% to 70% ethyl
acetate/hexanes afforded Cap-143, step a as a yellow solid (180 mg,
31%). R.sub.t=1.75 min (Cond.-MS-W1); 90% homogenity index; LC-MS:
Anal. Calc. for [M+H].sup.+ C.sub.13H.sub.14BrN.sub.2O: 293.03;
found: 293.04.
Cap-143
##STR00182##
[0401] To a cold (-60.degree. C.) solution of Cap-143, step a (154
mg, 0.527 mmol) in anhydrous tetrahydrofuran (5 mL) was added a
solution of n-butyllithium in hexanes (2.5 M, 0.25 mL, 0.633 mmol).
After 10 min, dry carbon dioxide was bubbled into the reaction
mixture for 10 min before it was quenched with 1N HCl and allowed
to warm to 25.degree. C. The mixture was then extracted with
dichloromethane (3.times.30 mL) and the combined organic extracts
were concentrated in vacuo. Purification of the residue by a
reverse phase HPLC (MeOH/water/TFA) afforded Cap-143 (16 mg, 12%).
R.sub.t=1.10 min (Cond.-MS-W1); 90% homogenity index; LC-MS: Anal.
Calc. for [M+H].sup.+ C.sub.14H.sub.15N.sub.2O.sub.3: 259.11;
found: 259.08.
Cap-144
##STR00183##
[0402] Cap-144, Step a
##STR00184##
[0404] 1,3-Dichloroisoquinoline (2.75 g, 13.89 mmol) was added in
small portions to a cold (0.degree. C.) solution of fuming nitric
acid (10 mL) and concentrated sulfuric acid (10 mL). The mixture
was stirred at 0.degree. C. for 0.5 h before it was gradually
warmed to 25.degree. C. where it stirred for 16 h. The mixture was
then poured into a beaker containing chopped ice and water and the
resulting suspension was stirred for 1 h at 0.degree. C. before it
was filtered to afford Cap-144, step a (2.73 g, 81%) as a yellow
solid which was used directly. R.sub.t=2.01 min. (Cond.-D1); 95%
homogenity index; LC-MS: Anal. Calc. for [M+H].sup.+
C.sub.9H.sub.5Cl.sub.2N.sub.2O.sub.2: 242.97; found: 242.92.
Cap-144, Step b
##STR00185##
[0406] Cap-144, step a (0.30 g, 1.23 mmol) was taken up in methanol
(60 mL) and treated with platinum oxide (30 mg), and the suspension
was subjected to Parr hydrogenation at 7 psi H.sub.2 for 1.5 h.
Then formalin (5 mL) and additional platinum oxide (30 mg) were
added, and the suspension was resubjected to Parr hydrogenation at
45 psi H.sub.2 for 13 h. It was then suction-filtered through
diatomaceous earth (CELITE.RTM.) and concentrated down to 1/4
volume. Suction-filtration of the ensuing precipitate afforded the
title compound as a yellow solid which was flash chromatographed on
silica gel eluting with 5% ethyl acetate in hexanes to 25% ethyl
acetate in hexanes to afford Cap-144, step b (231 mg, 78%) as a
pale yellow solid. R.sub.t=2.36 min (Cond.-D1); 95% homogenity
index; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.02 (s, 1H), 7.95
(d, J=8.6 Hz, 1H), 7.57-7.53 (m, 1H), 7.30 (d, J=7.3 Hz, 1H), 2.88
(s, 6H); LC-MS: Anal. Calc. for [M+H].sup.+
C.sub.11H.sub.11Cl.sub.2N.sub.2: 241.03; found: 241.02. HRMS: Anal.
Calc. for [M+H].sup.+ C.sub.11H.sub.11Cl.sub.2N.sub.2: 241.0299;
found: 241.0296.
Cap-144, Step c
##STR00186##
[0408] Cap-144, step c was prepared from Cap-144, step b according
to the procedure described for the preparation of Cap-139, step a.
R.sub.t=2.19 min (Cond.-D1); 95% homogenity index; LC-MS: Anal.
Calc. for [M+H].sup.+ C.sub.12H.sub.11ClN.sub.3: 232.06; found:
232.03. HRMS: Anal. Calc. for [M+H].sup.+
C.sub.12H.sub.11ClN.sub.3: 232.0642; found: 232.0631.
Cap-144
##STR00187##
[0410] Cap-144 was prepared according to the procedure described
for Cap-141. R.sub.t=2.36 min (Cond.-D1); 90%; LC-MS: Anal. Calc.
for [M+H].sup.+ C.sub.12H.sub.12ClN.sub.2O.sub.2: 238.01; found:
238.09.
Cap-145 to Cap-162
[0411] Cap-145 to Cap-162 were prepared from the appropriate
1-chloroisoquinolines according to the procedure described for the
preparation of Cap-138 (Method A) or Cap-139 (Method B) unless
noted otherwise as outlined below.
TABLE-US-00010 R.sub.t (LC-Cond.); % homogeneity Cap # Cap Method
Hydrolysis index; MS data Cap-145 ##STR00188## Prepared from
commercially available 1,3- dichloroisoquinoline B 12N HCl 1.14 min
(Cond.- MS-W1); 90% LC-MS: Anal. Calc. for [M + H].sup.+
C.sub.10H.sub.7ClNO.sub.2: 208.02; found: 208.00. Cap-146
##STR00189## Prepared from commercially available
3-hydroxyisoquinoline A 5N NaOH 1.40 min (Cond.- D1); 95%; LC- MS:
Anal. Calc. for [M + H].sup.+ C.sub.11H.sub.10NO.sub.3: 204.07;
found: 204.06. Cap-147 ##STR00190## Prepared from commercially
available 1-chloro-4- hydroxyisoquinoline B 5N NaOH 0.87 min
(Cond.- D1); 95%; LC- MS: Anal. Calc. for [M + H].sup.+
C.sub.11H.sub.10NO.sub.3: 204.07; found: 204.05. Cap-148
##STR00191## Prepared from commercially available
7-hydroxyisoquinoline A 5N NaOH 0.70 min (Cond.- D1); 95%; LC- MS:
Anal. Calc. for [M + H].sup.+ C.sub.11H.sub.10NO.sub.3: 204.07;
found: 204.05. Cap-149 ##STR00192## Prepared from commercially
available 5-hydroxyisoquinoline A 5N NaOH 0.70 min (Cond.- D1);
95%; LC- MS: Anal. Calc. for [M + H].sup.+
C.sub.11H.sub.10NO.sub.3: 204.07; found: 204.05. Cap 150
##STR00193## Prepared from 8- methoxy-1- chloroisoquinoline, which
can be synthesized following the procedure in WO 2003/099274 A 12N
HCl 0.26 min (Cond.- D1); 95%; LC- MS: Anal. Calc. for [M +
H].sup.+ C.sub.11H.sub.10NO.sub.3: 204.07; found: 204.04. Cap-151
3-chloro-5- methoxyisoquinoline- 1-carboxylic acid ##STR00194##
Prepared from 5- methoxy-1,3- dichloroisoquinoline, which can be
synthesized following the procedure in WO 2005/051410 B 12N HCl
1.78 min (Cond.- D1); 90%; LC- MS: Anal. Calc. for [M + H].sup.+
C.sub.11H.sub.9ClNO.sub.3: 238.03; found: 238.09. Cap-152
##STR00195## Prepared from commercially available 6-methoxy-1,3-
dichloroisoquinoline B 12N HCl 1.65 min (Cond.- D1); 95%; LC- MS:
Anal. Calc. for [M + H].sup.+ C.sub.11H.sub.9ClNO.sub.3: 238.00;
found: 238.09. Cap-153 ##STR00196## Prepared from 4-
bromoisoquinoline, which can be synthesized following the procedure
in WO 2003/062241 A 6N HCl 1.18 min (Cond.- MS-W1); 95%; LC-MS:
Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.7BrNO.sub.2: 251.97;
found: 251.95. Cap-154 ##STR00197## Prepared from 7-fluoro-
1-chloroisoquinoline, which can be synthesized following the
procedure in WO 2003/099274 B 5N NaOH 0.28 min (Cond.- MS-W1); 90%;
LC-MS: Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.7FNO.sub.2:
192.05; found: 192.03. Cap-155 ##STR00198## Prepared from 1,7-
dichloroisoquinoline, which can be synthesized following the
procedure in WO 2003/099274 B 5N NaOH 0.59 min (Cond.- MS-W1); 90%;
LC-MS: Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.7ClNO.sub.2:
208.02; found: 208.00. Cap-156 ##STR00199## Prepared from 1,6-
dichloroisoquinoline, which can be synthesized following the
procedure in WO 2003/099274 B 5N NaOH 0.60 min (Cond.- MS-W1); 90%;
LC-MS: Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.7ClNO.sub.2:
208.02; found: 208.03. Cap-157 ##STR00200## Prepared from 1,4-
dichloroisoquinoline, which can be synthesized following the
procedure in WO 2003/062241 B 12N HCl 1.49 min (Cond.- D1); 95%;
LC- MS: Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.17ClNO: 208.02;
found: 208.00. Cap-158 ##STR00201## Prepared from 1,5-
dichloroisoquinoline, which can be synthesized following the
procedure in WO 2003/099274 B 5N NaOH 0.69 min (Cond.- MS-W1); 90%;
LC-MS: Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.7ClNO.sub.2:
208.02; found: 208.01. Cap-159 ##STR00202## Prepared from 5-fluoro-
1-chloroisoquinoline, which can be synthesized following the
procedure in WO 2003/099274 B 5N NaOH 0.41 min (Cond.- MS-W1); 90%;
LC-MS: Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.7FNO.sub.2:
192.05; found: 192.03. Cap-160 ##STR00203## Prepared from 6-fluoro-
1-chloroisoquinoline, which can be synthesized following the
procedure in WO 2003/099274 B 5N NaOH 0.30 min (Cond.- MS-W1); 90%;
LC-MS: Anal. Calc. for [M + H].sup.+ C.sub.10H.sub.7FNO.sub.2:
192.05; found: 192.03. Cap-161 ##STR00204## Prepared from 4-
bromoquinoline-2- carboxylic acid and dimethylamine (DMSO,
100.degree. C.) -- --- 0.70 min (Cond. D1); 95%; LC- MS: Anal.
Calc. for [M + H].sup.+ C.sub.12H.sub.13N.sub.2O.sub.2: 217.10;
found: 217.06. Cap-162 ##STR00205## Prepared from m- anisidine
following the procedure described in J. Hetero. Chem., 17 (1993)
and Heterocycles, 60:953 (2003). -- -- 0.65 min (Cond.- M3); 95%;
LC- MS: Anal. Calc. for [M + H].sup.+ C.sub.11H.sub.10NO.sub.3:
204.07; found: 203.94.
Cap-163
##STR00206##
[0413] To a solution of 2-ketobutyric acid (1.0 g, 9.8 mmol) in
diethylether (25 ml) was added phenylmagnesium bromide (22 ml, 1M
in THF) dropwise. The reaction was stirred at .about.25.degree. C.
under nitrogen for 17.5 h. The reaction was acidified with 1N HCl
and the product was extracted with ethyl acetate (3.times.100 ml).
The combined organic layer was washed with water followed by brine
and dried over MgSO.sub.4. After concentration in vacuo, a white
solid was obtained. The solid was recrystallized from hexanes/ethyl
acetate to afford Cap-163 as white needles (883.5 mg). .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5 ppm, 500 MHz): 12.71 (br s, 1H),
7.54-7.52 (m, 2H), 7.34-7.31 (m, 2H), 7.26-7.23 (m, 1H), 5.52-5.39
(br s, 1H), 2.11 (m, 1H), 1.88 (m, 1H), 0.79 (app t, J=7.4 Hz,
3H).
Cap-164
##STR00207##
[0415] A mixture of 2-amino-2-phenylbutyric acid (1.5 g, 8.4 mmol),
formaldehyde (14 mL, 37% in water), 1N HCl (10 mL) and 10% Pd/C
(0.5 mg) in MeOH (40 mL) was exposed to H.sub.2 at 50 psi in a Parr
bottle for 42 h. The reaction was filtered over CELITE.RTM. and
concentrated in vacuo, the residue was taken up in MeOH (36 mL) and
the product was purified with a reverse phase HPLC
(MeOH/H.sub.2O/TFA) to afford the TFA salt of Cap-164 as a white
solid (1.7 g). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 500 MHz)
7.54-7.47 (m, 5H), 2.63 (m, 1H), 2.55 (s, 6H), 2.31 (m, 1H), 0.95
(app t, J=7.3 Hz, 3H).
Cap-165
##STR00208##
[0417] To a mixture of 2-amino-2-indanecarboxylic acid (258.6 mg,
1.46 mmol) and formic acid (0.6 ml, 15.9 mmol) in
1,2-dichloroethane (7 ml) was added formaldehyde (0.6 ml, 37% in
water). The mixture was stirred at .about.25.degree. C. for 15 min
then heated at 70.degree. C. for 8 h. The volatile component was
removed in vacuo, and the residue was dissolved in DMF (14 mL) and
purified by a reverse phase HPLC (MeOH/H.sub.2O/TFA) to afford the
TFA salt of Cap-165 as a viscous oil (120.2 mg). .sup.1H NMR
(DMSO-d.sub.6, .delta.=2.5 ppm, 500 MHz): 7.29-7.21 (m, 4H), 3.61
(d, J=17.4 Hz, 2H), 3.50 (d, J=17.4 Hz, 2H), 2.75 (s, 6H). LC-MS:
Anal. Calcd. for [M+H].sup.+ C.sub.12H.sub.16NO.sub.2: 206.12;
found: 206.07.
Cap-166a and Cap-166b
##STR00209##
[0419] Cap-166a and Cap-166b were prepared from
(1S,4S)-(+)-2-methyl-2,5-diazabicyclo[2.2.1]heptane (2HBr)
according to the method described for the synthesis of Cap-7a and
Cap-7b, with the exception that the benzyl ester intermediate was
separated using a semi-prep Chrialcel OJ column, 20.times.250 mm,
10 .mu.m eluting with 85:15 heptane/ethanol mixture at 10 mL/min
elution rate for 25 min. Cap-166b: .sup.1H NMR (DMSO-d.sub.6,
.delta.=2.5 ppm, 500 MHz): 7.45 (d, J=7.3 Hz, 2H), 7.27-7.19 (m,
3H), 4.09 (s, 1H), 3.34 (app br s, 1H), 3.16 (app br s, 1H), 2.83
(d, J=10.1 Hz, 1H), 2.71 (m, 2H), 2.46 (m, 1H), 2.27 (s, 3H), 1.77
(d, J=9.8 Hz, 1H), 1.63 (d, J=9.8 Hz, 1H). LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.14H.sub.19N.sub.2O.sub.2: 247.14; found:
247.11.
Cap-167
##STR00210##
[0421] A solution of racemic Boc-1,3-dihydro-2H-isoindole
carboxylic acid (1.0 g, 3.8 mmol) in 20% TFA/CH.sub.2Cl.sub.2 was
stirred at .about.25.degree. C. for 4 h. All the volatile component
was removed in vacuo. A mixture of the resultant crude material,
formaldehyde (15 mL, 37% in water), 1N HCl (10 mL) and 10% Pd/C (10
mg) in MeOH was exposed to H.sub.2 (40 PSI) in a Parr bottle for 23
h. The reaction mixture was filtered over CELITE.RTM. and
concentrated in vacuo to afford Cap-167 as a yellow foam (873.5
mg). .sup.1H NMR (DMSO-d.sub.6, .delta.=2.5 ppm, 500 MHz) 7.59-7.38
(m, 4H), 5.59 (s, 1H), 4.84 (d, J=14 Hz, 1H), 4.50 (d, J=14.1 Hz,
1H), 3.07 (s, 3H). LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.12NO.sub.2: 178.09; found: 178.65.
Cap-168
##STR00211##
[0423] Racemic Cap-168 was prepared from racemic
Boc-aminoindane-1-carboxylic acid according to the procedure
described for the preparation of Cap-167. The crude material was
employed as such.
Cap-169
##STR00212##
[0425] A mixture of 2-amino-2-phenylpropanoic acid hydrochloride
(5.0 g, 2.5 mmol), formaldehyde (15 ml, 37% in water), 1N HCl (15
ml), and 10% Pd/C (1.32 g) in MeOH (60 mL) was placed in a Parr
bottle and shaken under hydrogen (55 PSI) for 4 days. The reaction
mixture was filtered over CELITE.RTM. and concentrated in vacuo.
The residue was taken up in MeOH and purified by reverse phase
prep-HPLC (MeOH/water/TFA) to afford the TFA salt of Cap-169 as a
viscous semi-solid (2.1 g). .sup.1H NMR (CDCl.sub.3, .delta.=7.26
ppm, 500 MHz): 7.58-7.52 (m, 2H), 7.39-7.33 (m, 3H), 2.86 (br s,
3H), 2.47 (br s, 3H), 1.93 (s, 3H). LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.11H.sub.16NO.sub.2: 194.12; found: 194.12.
Cap-170
##STR00213##
[0426]
(S)-2-(Methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid
[0427] To (S)-2-amino-2-(tetrahydro-2H-pyran-4-yl)acetic acid (505
mg; 3.18 mmol; obtained from Astatech) in water (15 ml) was added
sodium carbonate (673 mg; 6.35 mmol), and the resultant mixture was
cooled to 0.degree. C. and then methyl chloroformate (0.26 ml; 3.33
mmol) was added dropwise over 5 minutes. The reaction was allowed
to stir for 18 hours while allowing the bath to thaw to ambient
temperature. The reaction mixture was then partitioned between 1N
HCl and ethyl acetate. The organic layer was removed and the
aqueous layer was further extracted with 2 additional portions of
ethyl acetate. The combined organic layers were washed with brine,
dried over magnesium sulfate, filtered and concentrated in vacuo to
afford Cap-170a colorless residue. .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. ppm 12.65 (1H, br s), 7.44 (1H, d, J=8.24
Hz), 3.77-3.95 (3H, m), 3.54 (3H, s), 3.11-3.26 (2H, m), 1.82-1.95
(1H, m), 1.41-1.55 (2H, m), 1.21-1.39 (2H, m); LC-MS: Anal. Calcd.
for [M+H].sup.+ C.sub.9H.sub.16NO.sub.5: 218.1; found 218.1.
Cap-171
##STR00214##
[0429] A solution of methyl
2-(benzyloxycarbonylamino)-2-(oxetan-3-ylidene)acetate (200 mg,
0.721 mmol; Il Farmaco (2001), 56, 609-613) in ethyl acetate (7 ml)
and CH.sub.2Cl.sub.2 (4.00 ml) was degassed by bubbling nitrogen
for 10 min. Dimethyl dicarbonate (0.116 ml, 1.082 mmol) and Pd/C
(20 mg, 0.019 mmol) were then added, the reaction mixture was
fitted with a hydrogen balloon and allowed to stir at ambient
temperature overnight at which time TLC (95:5
CH.sub.2Cl.sub.2/MeOH: visualized with stain made from 1 g
Ce(NH.sub.4).sub.2SO.sub.4, 6 g ammonium molybdate, 6 ml sulfuric
acid, and 100 ml water) indicated complete conversion. The reaction
was filtered through CELITE.RTM. and concentrated. The residue was
purified via BIOTAGE.RTM. (load with dichloromethane on 25 samplet;
elute on 25S column with dichloromethane for 3CV then 0 to 5%
MeOH/dichloromethane over 250 ml then hold at 5%
MeOH/dichloromethane for 250 ml; 9 ml fractions). Collected
fractions containing desired material and concentrated to 120 mg
(81%) of methyl 2-(methoxycarbonylamino)-2-(oxetan-3-yl)acetate as
a colorless oil. .sup.1H NMR (500 MHz, chloroform-d) .delta. ppm
3.29-3.40 (m, J=6.71 Hz, 1H) 3.70 (s, 3H) 3.74 (s, 3H) 4.55 (t,
J=6.41 Hz, 1H) 4.58-4.68 (m, 2H) 4.67-4.78 (m, 2H) 5.31 (br s, 1H).
LC-MS: Anal. Calcd. for [M+H].sup.+ C.sub.8H.sub.14NO.sub.5: 204.2;
found 204.0.
[0430] To methyl 2-(methoxycarbonylamino)-2-(oxetan-3-yl)acetate
(50 mg, 0.246 mmol) in THF (2 mL) and water (0.5 mL) was added
lithium hydroxide monohydrate (10.33 mg, 0.246 mmol). The resultant
solution was allowed to stir overnight at ambient temperature. TLC
(1:1 EA/Hex; Hanessian stain [1 g Ce(NH.sub.4).sub.2SO.sub.4, 6 g
ammonium molybdate, 6 ml sulfuric acid, and 100 ml water])
indicated .about.10% starting material remaining. Added an
additional 3 mg LiOH and allowed to stir overnight at which time
TLC showed no starting material remaining. Concentrated in vacuo
and placed on high vac overnight providing 55 mg lithium
2-(methoxycarbonylamino)-2-(oxetan-3-yl)acetate as a colorless
solid. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 3.39-3.47 (m, 1H)
3.67 (s, 3 H) 4.28 (d, J=7.93 Hz, 1H) 4.64 (t, J=6.26 Hz, 1H) 4.68
(t, J=7.02 Hz, 1H) 4.73 (d, J=7.63 Hz, 2H).
Cap-172
##STR00215##
[0431] Cap-172, Step a
##STR00216##
[0433] The following diazotization step was adapted from Barton, A.
et al., J. C. S. Perkin Trans I, 159-164 (1982): A solution of
NaNO.sub.2 (166 mg, 2.4 mmol) in water (0.6 mL) was added slowly to
a stirred, cold (0.degree. C.) solution of methyl
2-amino-5-ethyl-1,3-thiazole-4-carboxylate (186 mg, 1.0 mmol),
CuSO.sub.4.5H.sub.2O (330 mg, 1.32 mmol), NaCl (260 mg, 4.45 mmol)
and H.sub.2SO.sub.4 (5.5 mL) in water (7.5 mL). The mixture was
stirred at 0.degree. C. for 45 min and allowed to warm up to room
temperature where it stirred further for 1 h before CuCl (118 mg)
was added. This mixture was stirred further at room temperature for
16 h before it was diluted with brine and extracted with ether
twice. The organic layers were combined, dried over MgSO.sub.4 and
concentrated to give methyl 2-chloro-5-ethylthiazole-4-carboxylate
(i.e., Cap-172, step a) (175 mg, 85%) as an orange oil (80% pure)
which was used directly in the next reaction. R.sub.t=1.99 min
(Cond.-MD1); LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.7H.sub.9ClNO.sub.2S: 206.01; found: 206.05.
Cap-172
[0434] To a solution of methyl
2-chloro-5-ethylthiazole-4-carboxylate (175 mg) in
THF/H.sub.2O/MeOH (20 mL/3 mL/12 mL) was added LiOH (305 mg, 12.76
mmol). The mixture was stirred at room temperature overnight before
it was concentrated down and neutralized with 1N HCl in ether (25
mL). The residue was extracted twice with ethyl acetate and the
organic layers were combined, dried over MgSO.sub.4 and evaporated
to yield Cap-172 (60 mg, 74%) as a red solid which was used without
further purification. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
ppm 13.03-13.42 (1H, m), 3.16 (2H, q, J=7.4 Hz), 1.23 (3H, t, J=7.5
Hz). R.sub.t=1.78 min (Cond.-MD1); LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.6H.sub.7ClNO.sub.2S: 191.99; found: 191.99.
Cap-173
##STR00217##
[0435] Cap-173, Step a
##STR00218##
[0437] The following diazotization step was adapted from Barton, A.
et al., J.C.S. Perkin Trans I, 159-164 (1982): A solution of
NaNO.sub.2 (150 mg, 2.17 mmol) in water (1.0 mL) was added dropwise
to a stirred, cold (0.degree. C.) solution of methyl
2-amino-5-ethyl-1,3-thiazole-4-carboxylate (186 mg, 1.0 mmol) in
50% H.sub.3PO.sub.2 (3.2 mL). The mixture was stirred at 0.degree.
C. for 1 h and allowed to warm up to room temperature where it
stirred further for 2 h. After recooling to 0.degree. C., the
mixture was treated slowly with a solution of NaOH (85 mg) in water
(10 mL). The mixture was then diluted with saturated NaHCO.sub.3
solution and extracted twice with ether. The organic layers were
combined, dried over MgSO.sub.4 and concentrated to give methyl
5-ethylthiazole-4-carboxylate (i.e., Cap-173, step a) (134 mg, 78%)
as an orange oil (85% pure) which was used directly in the next
reaction. R.sub.t=1.58 min (Cond.-MD1); LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.7H.sub.10NO.sub.2S: 172.05; found: 172.05.
Cap-173
[0438] To a solution of methyl 5-ethylthiazole-4-carboxylate (134
mg) in THF/H.sub.2O/MeOH (18 mL/2.7 mL/11 mL) was added LiOH (281
mg, 11.74 mmol). The mixture was stirred at room temperature
overnight before it was concentrated down and neutralized with 1N
HCl in ether (25 mL). The residue was extracted twice with ethyl
acetate and the organic layers were combined, dried over MgSO.sub.4
and evaporated to yield Cap-173 (90 mg, 73%) as an orange solid
which was used without further purification. .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. ppm 12.74-13.04 (1H, m), 3.20 (2H, q, J=7.3
Hz), 1.25 (3H, t, J=7.5 Hz). R.sub.t=1.27 min (Cond.-MD1); LC-MS:
Anal. Calcd. for [M+H].sup.+ C.sub.6H.sub.8NO.sub.2S: 158.03;
found: 158.04.
Cap-174
##STR00219##
[0439] Cap-174, Step a
##STR00220##
[0441] Triflic anhydride (5.0 g, 18.0 mmol) was added dropwise to a
cold (0.degree. C.) solution of methyl 3-hydroxypicolinate (2.5 g,
16.3 mmol) and TEA (2.5 mL, 18.0 mmol) in CH.sub.2Cl.sub.2 (80 mL).
The mixture was stirred at 0.degree. C. for 1 h before it was
allowed to warm up to room temperature where it stirred for an
additional 1 h. The mixture was then quenched with saturated
NaHCO.sub.3 solution (40 mL) and the organic layer was separated,
washed with brine, dried over MgSO.sub.4 and concentrated to give
methyl 3-(trifluoromethylsulfonyloxy)picolinate (i.e., Cap-174,
step a) (3.38 g, 73%) as a dark brown oil (>95% pure) which was
used directly without further purification. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. ppm 8.72-8.79 (1H, m), 7.71 (1H, d, J=1.5 Hz),
7.58-7.65 (1H, m), 4.04 (3H, s). R.sub.t=1.93 min (Cond.-MD1);
LC-MS: Anal. Calcd. for [M+H].sup.+ C.sub.8H.sub.2F.sub.3NO.sub.5S:
286.00; found: 286.08.
Cap-174
[0442] To a solution of methyl
3-(trifluoromethylsulfonyloxy)picolinate (570 mg, 2.0 mmol) in DMF
(20 mL) was added LiCl (254 mg, 6.0 mmol), tributyl(vinyl)stannane
(761 mg, 2.4 mmol) and bis(triphenylphosphine)palladium dichloride
(42 mg, 0.06 mmol). The mixture was heated at 100.degree. C.
overnight before a saturated solution of KF (20 mL) was added to
the reaction mixture at room temperature. This mixture was stirred
for 4 h before it was filtered through CELITE.RTM. and the pad of
CELITE.RTM. was washed with ethyl acetate. The aqueous phase of the
filtrate was then separated and concentrated down in vacuo. The
residue was treated with 4N HCl in dioxanes (5 mL) and the
resulting mixture was extracted with methanol, filtered and
evaporated to afford Cap-174 (260 mg) as a green solid which was
slightly contaminated with inorganic salts but was used without
further purification. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
ppm 8.21 (1H, d, J=3.7 Hz), 7.81-7.90 (1H, m), 7.09 (1H, dd, J=7.7,
4.8 Hz), 6.98 (1H, dd, J=17.9, 11.3 Hz), 5.74 (1H, dd, J=17.9, 1.5
Hz), 5.20 (1H, d, J=11.0 Hz). R.sub.t=0.39 min (Cond.-MD1); LC-MS:
Anal. Calcd. for [M+H].sup.+ C.sub.8H.sub.8NO.sub.2: 150.06; found:
150.07.
Cap-175
##STR00221##
[0443] Cap-175, Step a
##STR00222##
[0445] To a solution of methyl
3-(trifluoromethylsulfonyloxy)picolinate (i.e., Cap-174, step a)
(570 mg, 2.0 mmol), an intermediate in the preparation of Cap-174,
in DMF (20 mL) was added LiCl (254 mg, 6.0 mmol),
tributyl(vinyl)stannane (761 mg, 2.4 mmol) and
bis(triphenylphosphine)palladium dichloride (42 mg, 0.06 mmol). The
mixture was heated at 100.degree. C. for 4 h before the solvent was
removed in vacuo. The residue was taken up in acetonitrile (50 mL)
and hexanes (50 mL) and the resulting mixture was washed twice with
hexanes. The acetonitrile layer was then separated, filtered
through CELITE.RTM., and evaporated. Purification of the residue by
flash chromatography on a Horizon instrument (gradient elution with
25% ethyl acetate in hexanes to 65% ethyl acetate in hexanes)
afforded methyl 3-vinylpicolinate (i.e., Cap-175, step a) (130 mg,
40%) as a yellow oil. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm
8.60 (1H, dd, J=4.6, 1.7 Hz), 7.94 (1H, d, J=7.7 Hz), 7.33-7.51
(2H, m), 5.72 (1H, d, J=17.2 Hz), 5.47 (1H, d, J=11.0 Hz), 3.99
(3H, s). R.sub.t=1.29 min (Cond.-MD1); LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.9H.sub.10NO.sub.2: 164.07; found: 164.06.
Cap-175, Step b
##STR00223##
[0447] Palladium on carbon (10%, 25 mg) was added to a solution of
methyl 3-vinylpicolinate (120 mg, 0.74 mmol) in ethanol (10 mL).
The suspension was stirred at room temperature under an atmosphere
of hydrogen for 1 h before it was filtered through CELITE.RTM. and
the pad of CELITE.RTM. was washed with methanol. The filtrate was
concentrated down to dryness to yield methyl 3-ethylpicolinate
(i.e., Cap-175, step b) which was taken directly into the next
reaction. R.sub.t=1.15 min (Cond.-MD1); LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.9H.sub.12NO.sub.2: 166.09; found: 166.09.
Cap-175
[0448] To a solution of methyl 3-ethylpicolinate in
THF/H.sub.2O/MeOH (5 mL/0.75 mL/3 mL) was added LiOH (35 mg, 1.47
mmol). The mixture was stirred at room temperature for 2 d before
additional LiOH (80 mg) was added. After an additional 24 h at room
temperature, the mixture was filtered and the solvent was removed
in vacuo. The residue was then treated with 4N HCl in dioxanes (5
mL) and the resulting suspension was concentrated down to dryness
to yield Cap-175 as a yellow solid which was used without further
purification. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm 8.47
(1H, dd, J=4.8, 1.5 Hz), 7.82-7.89 (1H, m), 7.53 (1H, dd, J=7.7,
4.8 Hz), 2.82 (2H, q, J=7.3 Hz), 1.17 (3H, t, J=7.5 Hz).
R.sub.t=0.36 min (Cond.-MD1); LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.8H.sub.10NO.sub.2: 152.07; found: 152.10.
Cap-176
##STR00224##
[0449]
(S)-2-(4,4-Difluorocyclohexyl)-2-(methoxycarbonylamino)acetic
acid
Cap-176, Step a
##STR00225##
[0451] A solution of 1,4-dioxaspiro[4.5]decan-8-one (15 g, 96 mmol)
in EtOAc (150 mL) was added to a solution of methyl
2-(benzyloxycarbonylamino)-2-(dimethoxyphosphoryl)acetate (21.21 g,
64.0 mmol) in 1,1,3,3-tetramethylguanidine (10.45 mL, 83 mmol) and
EtOAc (150 mL). The resulting solution was the stirred at ambient
temperature for 72 h and then it was diluted with EtOAc (25 mL).
The organic layer was washed with 1N HCl (75 mL), H.sub.2O (100 mL)
and brine (100 mL), dried (MgSO.sub.4), filtered and concentrated.
The residue was purified via BIOTAGE.RTM. (5% to 25% EtOAc/Hexanes;
300 g column). The combined fractions containing the product were
then concentrated under vacuum and the residue was re-crystallized
from hexanes/EtOAc to give white crystals that corresponded to
methyl
2-(benzyloxycarbonylamino)-2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate
(6.2 g) .sup.1H NMR (400 MHz, CDCl.sub.3-d) .delta. ppm 7.30-7.44
(5H, m), 6.02 (1H, br. s.), 5.15 (2H, s), 3.97 (4H, s), 3.76 (3H,
br. s.), 2.84-2.92 (2H, m), 2.47 (2H, t, J=6.40 Hz), 1.74-1.83 (4H,
m). LC (Cond. OL1): R.sub.t=2.89 min. LC-MS: Anal. Calcd. for
[M+Na].sup.+ C.sub.19H.sub.23NNaO.sub.6: 745.21; found: 745.47.
Cap-176, Step b
##STR00226##
[0453] Ester Cap-176, step b was prepared from alkene Cap-176, step
a according to the method of Burk, M. J. et al. (J. Am. Chem. Soc.,
117:9375-9376 (1995)) and references therein): A 500 mL
high-pressure bottle was charged with alkene Cap-176, step a (3.5
g, 9.68 mmol) in degassed MeOH (200 mL) under a blanket of N.sub.2.
The solution was then charged with
(-)-1,2-Bis((2S,5S)-2,5-dimethylphospholano)ethane(cyclooctadiene)
rhodium (I) tetrafluoroborate (0.108 g, 0.194 mmol) and the
resulting mixture was flushed with N.sub.2 (3.times.) and charged
with H.sub.2 (3.times.). The solution was shaken vigorously under
70 psi of H.sub.2 at ambient temperature for 72 h. The solvent was
removed under reduced pressure and the remaining residue was taken
up in EtOAc. The brownish solution was then filtered through a plug
of Silica Gel and eluted with EtOAc. The solvent was concentrated
under vacuum to afford a clear oil corresponding to ester Cap-176,
step b (3.4 g). .sup.1H NMR (500 MHz, CDCl.sub.3-d) .delta. ppm
7.28-7.43 (5H, m), 5.32 (1H, d, J=9.16 Hz), 5.06-5.16 (2H, m), 4.37
(1H, dd, J=9.00, 5.04 Hz), 3.92 (4H, t, J=3.05 Hz), 3.75 (3H, s),
1.64-1.92 (4H, m), 1.37-1.60 (5H, m). LC (Cond. OL1): R.sub.t=1.95
min. LC-MS: Anal. Calcd. for [M+H].sup.+C.sub.19H.sub.26NO.sub.6:
364.18; found: 364.27.
Cap-176, Step c
##STR00227##
[0455] Ester Cap-176, step b (4.78 g, 13.15 mmol) was dissolved in
THF (15 mL) followed by sequential addition of water (10 mL),
glacial acetic acid (26.4 mL, 460 mmol) and dichloroacetic acid
(5.44 mL, 65.8 mmol). The resulting mixture was stirred for 72 h at
ambient temperature, and the reaction was quenched by slow addition
of solid Na.sub.2CO.sub.3 with vigorous stirring until the release
of gas was no longer visible. Crude product was extracted into 10%
ethyl acetate-dichloromethane and the organic layers were combined,
dried (MgSO.sub.4) filtered and concentrated. The resulting residue
was purified via BIOTAGE.RTM. (0 to 30% EtOAc/Hex; 25 g column) to
afford ketone Cap-176, step c (3.86 g) as a clear oil. .sup.1H NMR
(400 MHz, CDCl.sub.3-d) .delta. ppm 7.28-7.41 (5H, m), 5.55 (1H, d,
J=8.28 Hz), 5.09 (2H, s), 4.46 (1H, dd, J=8.16, 5.14 Hz), 3.74 (3H,
s), 2.18-2.46 (5H, m), 1.96-2.06 (1H, m), 1.90 (1H, ddd, J=12.99,
5.96, 2.89 Hz), 1.44-1.68 (2H, m, J=12.36, 12.36, 12.36, 12.36,
4.77 Hz). LC (Cond. OL1): R.sub.t=1.66 min. LC-MS: Anal. Calcd. for
[M+Na].sup.+ C.sub.17H.sub.21NNaO.sub.5: 342.13; found: 342.10.
Cap-176, Step d
##STR00228##
[0457] DEOXO-FLUORO (3.13 mL, 16.97 mmol) was added to a solution
of ketone Cap-176, step c (2.71 g, 8.49 mmol) in CH.sub.2Cl.sub.2
(50 mL) followed by addition of a catalytic amount of EtOH (0.149
mL, 2.55 mmol). The resulting yellowish solution was stirred at rt
overnight. The reaction was quenched by addition of sat. aq.
NaHCO.sub.3 (25 mL) and the mixture was extracted with EtOAc
(3.times.75 mL)). The combined organic layers were dried
(MgSO.sub.4), filtered and dried to give a yellowish oil. The
residue was purified via BIOTAGEO chromatography (2% to 15%
EtOAc/Hex; 90 g column) and a white solid corresponding to the
difluoro amino acid difluoride Cap-176, step d (1.5 g) was
recovered. .sup.1H NMR (400 MHz, CDCl.sub.3-d) .delta. ppm
7.29-7.46 (5H, m), 5.34 (1H, d, J=8.28 Hz), 5.12 (2H, s), 4.41 (1H,
dd, J=8.66, 4.89 Hz), 3.77 (3H, s), 2.06-2.20 (2H, m), 1.83-1.98
(1H, m), 1.60-1.81 (4H, m), 1.38-1.55 (2H, m). .sup.19F NMR (376
MHz, CDCl.sub.3-d) .delta. ppm -92.15 (1F, d, J=237.55 Hz), -102.44
(1F, d, J=235.82 Hz). LC (Cond. OL1): R.sub.t=1.66 min. LC-MS:
Anal. Calcd. for [2M+Na].sup.+
C.sub.34H.sub.42F.sub.4N.sub.2NaO.sub.8: 705.28; found: 705.18.
Cap-176, Step e
##STR00229##
[0459] Difluoride Cap-176, step d (4 g, 11.72 mmol) was dissolved
in MeOH (120 mL) and charged with Pd/C (1.247 g, 1.172 mmol). The
suspension was flushed with N.sub.2 (3.times.) and the reaction
mixture was placed under 1 atm of H.sub.2 (balloon). The mixture
was stirred at ambient temperature for 48 h. The suspension was
then filtered though a plug of CELITE.RTM. and concentrated under
vacuum to give an oil that corresponded to amino acid Cap-176, step
e (2.04 g) and that was used without further purification. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 3.62 (3H, s), 3.20 (1H, d,
J=5.77 Hz), 1.91-2.09 (2H, m), 1.50-1.88 (7H, m), 1.20-1.45 (2H,
m). .sup.19F NMR (376 MHz, DMSO-d.sub.6) .delta. ppm -89.39 (1F, d,
J=232.35 Hz), -100.07 (1F, d, J=232.35 Hz). .sup.13C NMR (101 MHz,
DMSO-d.sub.6) .delta. ppm 175.51 (1C, s), 124.10 (1C, t, J=241.21,
238.90 Hz), 57.74 (1C, s), 51.39 (1C, s), 39.23 (1C, br. s.),
32.02-33.83 (2C, m), 25.36 (1C, d, J=10.02 Hz), 23.74 (1C, d,
J=9.25 Hz). LC (Cond. OL2): R.sub.t=0.95 min. LC-MS: Anal. Calcd.
for [2M+H].sup.+ C.sub.18H.sub.31F.sub.4N.sub.2O.sub.2: 415.22;
found: 415.40.
Cap-176, Step f
##STR00230##
[0461] Methyl chloroformate (260.495 mL, 19.30 mmol) was added to a
solution of amino acid Cap-176, step e (2 g, 9.65 mmol) and DIEA
(6.74 mL, 38.6 mmol) in CH.sub.2Cl.sub.2 (100 mL). The resulting
solution was stirred at rt for 3 h and volatiles were removed under
reduced pressure. The residue was purified via BIOTAGE.RTM. (0% to
20% EtOAc/Hex; 90 g column). A clear oil that solidified upon
standing under vacuum and corresponding to carbamate Cap-176, step
f (2.22 g) was recovered. .sup.1H NMR (500 MHz, CDCl.sub.3-d)
.delta. ppm 5.27 (1H, d, J=8.55 Hz), 4.39 (1H, dd, J=8.85, 4.88
Hz), 3.77 (3H, s), 3.70 (3H, s), 2.07-2.20 (2H, m), 1.84-1.96 (1H,
m), 1.64-1.82 (4H, m), 1.39-1.51 (2H, m). .sup.19F NMR (471 MHz,
CDCl.sub.3-d) .delta. ppm -92.55 (1F, d, J=237.13 Hz), -102.93 (1F,
d, J=237.12 Hz). .sup.13C NMR (126 MHz, CDCl.sub.3-d) .delta. ppm
171.97 (1C, s), 156.69 (1C, s), 119.77-125.59 (1C, m), 57.24 (1C,
br. s.), 52.48 (1C, br. s.), 52.43 (1C, s), 39.15 (1C, s),
32.50-33.48 (2C, m), 25.30 (1C, d, J=9.60 Hz), 24.03 (1C, d, J=9.60
Hz). LC (Cond. OL1): R.sub.t=1.49 min. LC-MS: Anal. Calcd. for
[M+Na].sup.+ C.sub.11H.sub.17F.sub.2NNaO.sub.4: 288.10; found:
288.03.
Cap-176
##STR00231##
[0462]
(S)-2-(4,4-Difluorocyclohexyl)-2-(methoxycarbonylamino)acetic
acid
[0463] A solution of LiOH (0.379 g, 15.83 mmol) in water (25 mL)
was added to a solution of carbamate Cap-176, step f (2.1 g, 7.92
mmol) in THF (75 mL) and the resulting mixture was stirred at
ambient temperature for 4 h. THF was removed under vacuum and the
remaining aqueous phase was acidified with 1N HCl solution (2 mL)
and then extracted with EtOAc (2.times.50 mL). The combined organic
layers were dried (MgSO.sub.4), filtered and concentrated to give a
white foam corresponding to Cap-176 (1.92 g). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.73 (1H, s), 7.50 (1H, d, J=8.78 Hz),
3.97 (1H, dd, J=8.53, 6.02 Hz), 3.54 (3H, s), 1.92-2.08 (2H, m),
1.57-1.90 (5H, m), 1.34-1.48 (1H, m), 1.27 (1H, qd, J=12.72, 3.26
Hz). .sup.19F NMR (376 MHz, DMSO-d.sub.6) .delta. ppm -89.62 (1F,
d, J=232.35 Hz), -99.93 (1F, d, J=232.35 Hz). LC (Cond. OL2):
R.sub.t=0.76 min. LC-MS: Anal. Calcd. for [M-H].sup.+
C.sub.10H.sub.14F.sub.2NO.sub.4: 250.09; found: 250.10.
Cap-177a and Cap-177b
##STR00232##
[0464] Cap-177a and Cap-177b, Step a
##STR00233##
[0466] 1,1,3,3-Tetramethylguanidine (0.985 mL, 7.85 mmol) was added
to a stirred solution of methyl
2-(benzyloxycarbonylamino)-2-(dimethoxyphosphoryl)acetate (2.0 g,
6.0 mmol) in EtOAc (40 mL) and the mixture was stirred at rt under
N.sub.2 for 10 min. Then dihydro-2H-pyran-3(4H)-one [23462-75-1]
(0.604 g, 6.04 mmol) was added and the mixture was stirred at rt
for 16 h. The reaction mixture was then cooled in freezer for 10
min and neutralized with aq. citric acid (1.5 g in 20 mL water).
The two phases were partitioned and the organic layer was washed
with 0.25 N aq.HCl and brine, and then dried (MgSO.sub.4) and
concentrated to a colorless oil. The crude material was purified by
flash silica chromatography (loading solvent: DCM, eluted with
EtOAc/Hexanes, gradient from 20% to 30% EtOAc) to yield two
isomeric products: The first eluted product was (Z)-methyl
2-(benzyloxycarbonylamino)-2-(2H-pyran-3(4H,5H,6H)-ylidene)acetate
(490 mg) (white solid), and the second was (E)-methyl
2-(benzyloxycarbonylamino)-2-(2H-pyran-3(4H,5H,6H)-ylidene)acetate
(433 mg) (white solid). LC-MS retention time 1.398 min (for
Z-isomer) and 1.378 min (for E-isomer); m/z 304.08 (for Z-isomer)
and 304.16 (for E-isomer) (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) (for Z-isomer) .delta. ppm
7.30-7.44 (m, 5H), 6.18 (br. s., 1H), 5.10-5.17 (m, 2H), 4.22 (s,
2H), 3.78 (br. s., 3H), 2.93-3.02 (m, 2H), 1.80 (dt, J=11.7, 5.8
Hz, 2H), 1.62 (s, 2H). .sup.1H NMR (400 MHz, chloroform-d) (for
E-isomer) .delta. ppm 7.31-7.44 (m, 5H), 6.12 (br. s., 1H),
5.13-5.17 (m, 2H), 4.64 (br. s., 2H), 3.70-3.82 (m, 5H), 2.49 (t,
J=6.5 Hz, 2H), 1.80 (br. s., 2H). (Note: the absolute
regiochemistry was determined by .sup.1H NMR shifts and coupling
constants).
Cap-177a and Cap-177b, Step b
##STR00234##
[0468]
(-)-1,2-Bis((2S,5S)-2,5-dimethylphospholano)ethane(cyclooctadiene)--
rhodium(I)tetrafluoroborate (28.2 mg, 0.051 mmol) was added to a
stirred solution of (Z)-methyl
2-(benzyloxycarbonylamino)-2-(2H-pyran-3(4H,5H,6H)-ylidene)acetate
(310 mg, 1.015 mmol) in MeOH (10 mL) and the mixture was vacuum
flushed with N.sub.2, followed by H.sub.2, and then the reaction
was stirred under H.sub.2 (60 psi) at rt for 2 d. The reaction
mixture was concentrated and the residue was purified by flash
silica chromatography (loading solvent: DCM, eluted with 20% EtOAc
in hexanes) to yield (S)-methyl
2-(benzyloxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)acetate
(204 mg) as clear colorless oil. LC-MS retention time 1.437 min;
m/z 307.89 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 7.30-7.46 (m, 5H),
5.32 (d, J=8.8 Hz, 1H), 5.12 (s, 2H), 4.36 (dd, J=8.9, 5.6 Hz, 1H),
3.84-3.98 (m, 2H), 3.77 (s, 3H), 3.28-3.37 (m, 1H), 3.23 (dd,
J=11.3, 10.5 Hz, 1H), 2.04-2.16 (m, 1H), 1.61-1.75 (m, 3H),
1.31-1.43 (m, 1H).
[0469] The other stereoisomer ((E)-methyl
2-(benzyloxycarbonylamino)-2-(2H-pyran-3(4H,5H,6H)-ylidene)acetate)
(360 mg, 1.18 mmol) was reduced in a similar manner to yield
(S)-methyl
2-(benzyloxycarbonylamino)-2-((R)-tetrahydro-2H-pyran-3-yl)acetate
(214 mg) as clear colorless oil. LC-MS retention time 1.437 min;
m/z 308.03 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 7.30-7.44 (m, 5H),
5.31 (d, J=9.0 Hz, 1H), 5.12 (s, 2H), 4.31 (dd, J=8.7, 6.9 Hz, 1H),
3.80-3.90 (m, 2H), 3.77 (s, 3H), 3.37 (td, J=10.8, 3.5 Hz, 1H),
3.28 (dd, J=11.3, 9.8 Hz, 1H), 1.97-2.10 (m, 1H), 1.81 (d, J=11.5
Hz, 1H), 1.61-1.72 (m, 2H), 1.33-1.46 (m, 1H).
Cap-177a and Cap-177b, Step c
##STR00235##
[0471] 10% Pd/C (69.3 mg, 0.065 mmol) was added to a solution of
(S)-methyl
2-(benzyloxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)acetate
(200 mg, 0.651 mmol) and dimethyl dicarbonate [4525-33-1] (0.104
mL, 0.976 mmol) in MeOH (10 mL). The reaction mixture was vacuum
flushed with N.sub.2, followed by H.sub.2, and then the reaction
was stirred under H.sub.2 (55 psi) at rt for 5 h. The reaction
mixture was filtered through CELITE.RTM./silica pad and the
filtrate was concentrated to a colorless oil. The crude oil was
purified by flash silica chromatography (loading solvent: DCM,
eluted with 30% EtOAc in hexanes) to yield product (S)-methyl
2-(methoxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)acetate
(132 mg) as colorless oil. LC-MS retention time 0.92 min; m/z
231.97 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 5.24 (d, J=8.5 Hz,
1H), 4.34 (dd, J=8.9, 5.6 Hz, 1H), 3.84-3.97 (m, 2H), 3.77 (s, 3H),
3.70 (s, 3H), 3.29-3.38 (m, 1H), 3.23 (dd, J=11.2, 10.4 Hz, 1H),
2.03-2.14 (m, 1H), 1.56-1.75 (m, 3H), 1.32-1.43 (m, 1H).
[0472] Another diastereomer ((S)-methyl
2-(benzyloxycarbonylamino)-2-((R)-tetrahydro-2H-pyran-3-yl)acetate)
was transformed in a similar manner to yield (S)-methyl
2-(methoxycarbonylamino)-2-((R)-tetrahydro-2H-pyran-3-yl)acetate as
clear colorless oil. LC-MS retention time 0.99 min; m/z 231.90
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 5.25 (d, J=8.0 Hz,
1H), 4.29 (dd, J=8.4, 7.2 Hz, 1H), 3.82-3.90 (m, 2H), 3.77 (s, 3H),
3.70 (s, 3H), 3.37 (td, J=10.8, 3.3 Hz, 1H), 3.28 (t, J=10.5 Hz,
1H), 1.96-2.08 (m, 1H), 1.81 (dd, J=12.9, 1.6 Hz, 1H), 1.56-1.72
(m, 2H), 1.33-1.46 (m, 1H).
Cap-177a and Cap-177b, step d
##STR00236##
[0474] To a solution of (S)-methyl
2-(methoxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)acetate
(126 mg, 0.545 mmol) in THF (4 mL) stirring at rt was added a
solution of 1M LiOH (1.090 mL, 1.090 mmol) in water. The reaction
was stirred at rt for 3 h, neutralized with 1M HCl (1.1 mL) and
extracted with EtOAc (3.times.10 mL). The organics were dried,
filtered and concentrated to yield
(S)-2-(methoxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)acetic
acid (Cap-177a) (125 mg) as a clear colorless oil. LC-MS retention
time 0.44 min; m/z 218.00 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 5.28 (d, J=8.8 Hz,
1H), 4.38 (dd, J=8.7, 5.6 Hz, 1H), 3.96-4.04 (m, 1H), 3.91 (d,
J=11.0 Hz, 1H), 3.71 (s, 3H), 3.33-3.41 (m, 1H), 3.24-3.32 (m, 1H),
2.10-2.24 (m, 1H), 1.74-1.83 (m, 1H), 1.63-1.71 (m, 2H), 1.35-1.49
(m, 1H).
[0475] Another diastereomer ((S)-methyl
2-(methoxycarbonylamino)-2-((R)-tetrahydro-2H-pyran-3-yl)acetate)
was transformed in a similar manner to yield
(S)-2-(methoxycarbonylamino)-2-((R)-tetrahydro-2H-pyran-3-yl)acetic
acid (Cap-177b) as clear colorless oil. LC-MS retention time 0.41
min; m/z 217.93 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 6.18 (br. s., 1H),
5.39 (d, J=8.5 Hz, 1H), 4.27-4.37 (m, 1H), 3.82-3.96 (m, 2H), 3.72
(s, 3H), 3.42 (td, J=10.8, 3.3 Hz, 1H), 3.35 (t, J=10.4 Hz, 1H),
2.01-2.18 (m, 1H), 1.90 (d, J=11.8 Hz, 1H), 1.59-1.76 (m, 2H),
1.40-1.54 (m, 1H).
Cap-178
##STR00237##
[0476] Cap-178, Step a
##STR00238##
[0478] To a solution of
(2S,3S,4S)-2-methyl-3,4-dihydro-2H-pyran-3,4-diyl diacetate (5 g,
23.34 mmol) in 20 mL of MeOH in a hydrogenation tank was added Pd/C
(150 mg, 0.141 mmol). The resulting mixture was hydrogenated at 40
psi on Parr Shaker for 1 hour. The mixture was then filtered and
the filtrate was concentrated to afford Cap-178, step a (5.0 g) as
a clear oil, which solidified while standing. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 4.85-4.94 (1H, m), 4.69 (1H, t, J=9.46 Hz),
3.88-3.94 (1H, m), 3.44 (1H, td, J=12.21, 1.83 Hz), 3.36 (1H, dq,
J=9.42, 6.12 Hz), 2.03-2.08 (1H, m), 2.02 (3H, s), 2.00 (3H, s),
1.70-1.80 (1H, m), 1.16 (3H, d, J=6.10 Hz).
Cap-178, Step b
##STR00239##
[0480] To a solution of Cap-178, step a (5.0 g, 23 mmol) in 50 mL
of MeOH was added several drops of sodium methoxide. After stirring
at room temperature for 30 min, sodium methoxide (0.1 mL, 23.12
mmol) was added and the solution was stirred at room temperature
overnight. The solvent was then removed under vacuum. The residue
was diluted with benzene and concentrated to afford the
corresponding diol as a yellow solid.
[0481] The solid was dissolved in 50 mL of pyridine and to this
solution at -35.degree. C. was added benzoyl chloride (2.95 mL,
25.4 mmol) dropwise. The resulting mixture was stirred at
-35.degree. C. for 1 hour then at room temperature overnight. The
mixture was diluted with Et.sub.2O and washed with water. The
aqueous layer was extracted with EtOAc (2.times.). The combined
organic layers were dried with MgSO.sub.4 and concentrated. The
crude product was purified by flash chromatography (silica gel,
5%-15% EtOAc/Hex) to afford Cap-178, step b (4.5 g) as clear oil
which slowly crystallized upon prolonged standing. LC-MS: Anal.
Calcd. for [M+Na].sup.+ C.sub.13H.sub.16NaO.sub.4 259.09; found
259.0; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.02-8.07 (2H,
m), 7.55-7.61 (1H, m), 7.45 (2H, t, J=7.78 Hz), 5.01 (1 H, ddd,
J=11.44, 8.70, 5.49 Hz), 3.98 (1H, ddd, J=11.90, 4.88, 1.53 Hz),
3.54 (1H, td, J=12.36, 2.14 Hz), 3.41 (1H, t, J=9.00 Hz), 3.31-3.38
(1H, m), 2.13-2.19 (1H, m), 1.83-1.94 (1H, m), 1.36 (3H, d, J=5.80
Hz).
Cap-178, Step c
##STR00240##
[0483] To a mixture of NaH (1.143 g, 28.6 mmol) (60% in mineral
oil) in 6 mL of CS.sub.2 was added Cap-178, step b (4.5 g, 19 mmol)
in 40 mL of CS.sub.2 dropwise over 15 min. The resulting mixture
was stirred at room temperature for 30 min. The mixture turned
light orange with some solid. MeI (14.29 mL, 229 mmol) was then
added dropwise over 20 min. The mixture was then stirred at room
temperature overnight. The reaction was carefully quenched with
saturated NH.sub.4Cl solution. The mixture was extracted with EtOAc
(3.times.). The combined organic layers were dried with MgSO.sub.4
and concentrated. The crude product was purified by flash
chromatography (silica gel, 6% EtOAc/Hex) to afford Cap-178, step c
(3.13 g) as clear oil. LC-MS: Anal. Calcd. for [M+Na].sup.+
C.sub.15H.sub.18NaO.sub.4S.sub.2 349.05; found 349.11; .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 7.94-8.00 (2 H, m), 7.50-7.58
(1H, m), 7.41 (2H, t, J=7.78 Hz), 5.96 (1H, t, J=9.46 Hz), 5.28
(1H, ddd, J=11.37, 9.38, 5.49 Hz), 4.02 (1H, ddd, J=11.98, 4.96,
1.68 Hz), 3.54-3.68 (2H, m), 2.48 (3H, s), 2.31 (1H, dd), 1.88-1.99
(1H, m), 1.28 (3H, d).
Cap-178, Step d
##STR00241##
[0485] To a mixture of Cap-178, step c (3.13 g, 9.59 mmol) and AIBN
(120 mg, 0.731 mmol) in 40 mL of benzene at 80.degree. C. was added
tri-n-butyltin hydride (10.24 mL, 38.4 mmol). The resulting mixture
was stirred at reflux temperature for 20 min then cooled to room
temperature. The mixture was diluted with diethyl ether and 100 mL
of KF (10 g) aqueous solution was added and the mixture was stirred
vigorously for 30 min. The two layers were then separated and the
aqueous phase was extracted with EtOAc (2.times.). The organic
layer was dried with MgSO.sub.4 and concentrated. The crude product
was purified by flash chromatography (silica gel, deactivated with
3% Et.sub.3N in Hexanes and flushed with 3% Et.sub.3N in Hexanes to
remove tributyltin derivative and then eluted with 15% EtOAc/Hex)
to afford Cap-178, step d (1.9 g) as clear oil. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 7.98-8.07 (2H, m), 7.52-7.58 (1H, m),
7.43 (2H, t, J=7.63 Hz), 5.08-5.17 (1H, m), 4.06 (1H, ddd, J=11.90,
4.88, 1.53 Hz), 3.50-3.59 (2H, m), 2.08-2.14 (1H, m), 1.99-2.06
(1H, m), 1.69-1.80 (1H, m), 1.41-1.49 (1H, m), 1.24 (3H, d, J=6.10
Hz).
Cap-178, Step e
##STR00242##
[0487] To a mixture of Cap-178, step d (1.9 g, 8.63 mmol) in 10 mL
of MeOH was added sodium methoxide (2 mL, 4.00 mmol) (2 M in
methanol). The resulting mixture was stirred at room temperature
for 5 hours. The solvent was removed under vacuum. The mixture was
neutralized with saturated NH.sub.4Cl solution and extracted with
EtOAc (3.times.). The organic layers were dried with MgSO.sub.4 and
concentrated to afford Cap-178, step e (0.8 g) as clear oil. The
product was used in the next step without further purification.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 4.01 (1H, ddd,
J=11.80, 5.02, 1.76 Hz), 3.73-3.83 (1H, m), 3.36-3.46 (2H, m),
1.92-2.00 (1H, m), 1.88 (1H, m), 1.43-1.56 (1H, m), 1.23 (3H, d),
1.15-1.29 (1H, m).
Cap-178, Step f
##STR00243##
[0489] Tosyl-Cl (2.63 g, 13.77 mmol) was added to a solution of
Cap-178, step e (0.8 g, 6.89 mmol) and pyridine (2.23 mL, 27.5
mmol) in 100 mL of CH.sub.2Cl.sub.2. The resulting mixture was
stirred at room temperature for 3 days. 10 mL of water was then
added into the reaction mixture and the mixture was stirred at room
temperature for an hour. The two layers were separated and the
organic phase was washed with water and 1 N HCl aq. solution. The
organic phase was dried with MgSO.sub.4 and concentrated to afford
Cap-178, step f (1.75 g) as a light yellow solid. The product was
used in the next step without further purification. Anal. Calcd.
for [M+H].sup.+ C.sub.13H.sub.19O.sub.4S 271.10; found 270.90;
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 7.79 (2H, d, J=8.24
Hz), 7.34 (2H, d, J=7.93 Hz), 4.53-4.62 (1H, m), 3.94 (1H, ddd,
J=12.13, 4.96, 1.83 Hz), 3.29-3.41 (2H, m), 2.45 (3H, s), 1.90-1.97
(1H, m), 1.79-1.85 (1H, m), 1.64-1.75 (1H, m), 1.38-1.48 (1H, m),
1.17 (3H, d, J=6.10 Hz).
Cap-178, Step g
##STR00244##
[0491] To a microwave tube was placed ethyl
2-(diphenylmethyleneamino)acetate (1.6 g, 5.92 mmol) and Cap-178,
step f (1.6 g, 5.92 mmol). 10 mL of toluene was added. The tube was
sealed and LiHMDS (7.1 mL, 7.10 mmol) (1 N in toluene) was added
dropwise under N.sub.2. The resulting dark brown solution was
heated at 100.degree. C. under microwave radiation for 6 hours. To
the mixture was then added water and the mixture was extracted with
EtOAc (3.times.). The combined organic layers were washed with
brine, dried with MgSO.sub.4 and concentrated to afford a
diastereomeric mixture of Cap-3, step g (3.1 g) as an orange oil.
The crude mixture was submitted to the next step without
separation. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.23H.sub.28NO.sub.3 366.21; found 366.3.
Cap-178, Step h
##STR00245##
[0493] To a solution of the diastereomeric mixture of ethyl
Cap-178, step g in 20 mL of THF was added HCl (30 ml, 60.0 mmol) (2
N aqueous). The resulting mixture was stirred at room temperature
for 1 hour. The mixture was extracted with EtOAc and the aqueous
layer was concentrated to afford an HCl salt of Cap-178, step h
(1.9 g) as an orange oil. The salt was used in the next step
without further purification. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.20NO.sub.3 202.14; found 202.1.
Cap-178, Step i
##STR00246##
[0495] A solution of 1.9 g Cap-178, step h (HCl salt), DiPEA (4.19
mL, 24.0 mmol) and methyl chloroformate (1.24 mL, 16.0 mmol) in 20
mL of CH.sub.2Cl.sub.2 was stirred at room temperature for 1 hour.
The mixture was diluted with CH.sub.2Cl.sub.2 and washed with
water. The organic layer was dried with Na.sub.2SO.sub.4 and
concentrated. The crude product was purified by flash
chromatography (silica gel, 0-20% EtOAc/Hex) to afford Cap-178,
step (1.1 g) as a yellow oil. Anal. Calcd. for [M+Na].sup.+
C.sub.12H.sub.21NNaO.sub.5 282.13; found 282.14; .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 5.16 (1H, br. s.), 4.43-4.58 (1H, m),
4.17-4.28 (2H, m), 3.89-4.03 (1H, m), 3.72-3.78 (2H, m), 3.67-3.72
(3H, m), 2.07-2.19 (1H, m), 1.35-1.77 (4H, m), 1.30 (3H, td,
J=7.09, 2.89 Hz), 1.19 (3H, d, J=6.53 Hz).
Cap-178, Step j
##STR00247##
[0497] To a mixture of Cap-178, step i (1.1 g, 4.2 mmol) in 5 mL of
THF and 2 mL of water was added LiOH (6.36 mL, 12.7 mmol) (2 N
aq.). The resulting mixture was stirred at room temperature
overnight. The mixture was then neutralized with 1 N HCl aq. and
extracted with EtOAc (3.times.). The combined organic layers were
dried with MgSO.sub.4 and concentrated to afford Cap-178, step j
(0.8 g) as a clear oil. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.18NO.sub.5 232.12; found 232.1; .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 5.20 (1H, d, J=8.28 Hz), 4.54 (1H, t,
J=8.16 Hz), 3.95-4.10 (1H, m), 3.66-3.85 (5H, m), 2.15-2.29 (1H,
m), 1.41-1.85 (4H, m), 1.23 (3H, dd, J=6.53, 1.76 Hz).
Cap-178, Step k
##STR00248##
[0499] To a solution of Cap-178, step j (240 mg, 1.04 mmol),
(S)-1-phenylethanol (0.141 mL, 1.142 mmol) and EDC (219 mg, 1.14
mmol) in 10 mL of CH.sub.2Cl.sub.2 was added DMAP (13.95 mg, 0.114
mmol). The resulting solution was stirred at room temperature
overnight and the solvent was removed under vacuum. The residue was
taken up into EtOAc, washed with water, dried with MgSO.sub.4 and
concentrated. The crude product was purified by chromatography
(silica gel, 0-15% EtOAc/Hexanes) to afford Cap-178, step k as a
mixture of two diastereomers. The mixture was separated by chiral
HPLC (CHIRALPAK.RTM. AS column, 21.times.250 mm, 10 um) eluting
with 90% 0.1% diethylamine/Heptane-10% EtOH at 15 mL/min to afford
Cap-178, step k stereoisomer 1 (eluted first) and Cap-178, step k
stereoisomer 2 (eluted second) as white solids. The stereochemistry
of the isomers was not assigned.
[0500] Cap-178, step k stereoisomer 1 (130 mg): LC-MS: Anal. Calcd.
for [M+Na].sup.+ C.sub.18H.sub.25NNaO.sub.5 358.16; found 358.16;
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 7.28-7.38 (5 H, m),
5.94 (1H, q, J=6.71 Hz), 5.12 (1H, d, J=9.16 Hz), 4.55 (1H, t,
J=9.00 Hz), 3.72-3.81 (1H, m), 3.67 (3H, s), 3.60-3.70 (2H, m),
1.98-2.08 (1H, m), 1.59 (3H, d, J=6.71 Hz), 1.38-1.47 (2H, m), 1.30
(2H, t, J=5.34 Hz), 0.93 (3H, d, J=6.41 Hz).
Cap-178, Stereoisomer 1
[0501] To a solution of Cap-178, step k stereoisomer 1
((S)-2-(methoxycarbonylamino)-2-((2S,4R)-2-methyltetrahydro-2H-pyran-4-yl-
)acetic acid) (150 mg, 0.447 mmol) in 10 mL of EtOH was added Pd/C
(20 mg, 0.188 mmol) and the mixture was hydrogenated on Parr shaker
at 40 psi overnight. The mixture was then filtered and the filtrate
was concentrated to afford Cap-178, stereoisomer 1 (100 mg) as a
sticky white solid. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.18NO.sub.5 232.12; found 232.1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 5.14-5.27 (1H, m), 4.51 (1H, t, J=8.39 Hz),
3.90-4.07 (1H, m), 3.60-3.83 (5H, m), 2.06-2.27 (1H, m), 1.45-1.77
(4H, m), 1.21 (3H, d, J=6.41 Hz).
Cap-179 (Enantiomer-1 and Enantiomer-2)
##STR00249##
[0502] Cap-179, Step a
##STR00250##
[0504] 2,6-Dimethyl-4H-pyran-4-one (10 g, 81 mmol) was dissolved in
ethanol (125 mL) and Pd/C (1 g, 0.94 mmol) was added. The mixture
was hydrogenated in a Parr shaker under H.sub.2 (0.325 g, 161 mmol)
(70 psi) at room temperature for 12 hrs. The catalyst was filtered
through a pad of CELITE.RTM. and washed with ethanol. The filtrate
was concentrated in vacuum and the residue was purified via
BIOTAGE.RTM. (2% to 25% EtOAc/Hex; 160 g column). Two fractions of
clear oils were isolated. The first eluting one corresponded to
(2R,6S)-2,6-dimethyldihydro-2H-pyran-4(3H)-one (1.8 g) while the
second one corresponded to Cap-179, step a (1.8 g).
(2R,6S)-2,6-Dimethyldihydro-2H-pyran-4(3H)-one data: .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 3.69 (2H, ddd, J=11.29, 5.95,
2.29 Hz), 2.24-2.36 (2H, m), 2.08-2.23 (2H, m), 1.18-1.34 (6H, m);
.sup.13C NMR (126 MHz, CDCl.sub.3) .delta. ppm 206.96 (1C, br. s.),
72.69 (2C, s), 48.70 (2C, s), 21.72 (2C, s).
[0505] Cap-179, step a data: .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. ppm 3.69-3.78 (1H, m), 3.36-3.47 (2H, m), 2.10 (1H, br.
s.), 1.88 (2H, dd, J=12.05, 4.73 Hz), 1.19 (6H, d, J=6.10 Hz), 1.10
(2H, q, J=10.70 Hz); .sup.13C NMR (126 MHz, CDCl.sub.3) .delta. ppm
71.44 (2C, s), 67.92 (1C, s), 42.59 (2C, s), 21.71 (2C, s).
Cap-179, Step b
##STR00251##
[0507] DEAD (2.311 mL, 14.59 mmol) was added drop wise to a
solution of Cap-179, step a (0.38 g, 2.92 mmol), 4-nitrobenzoic
acid (2.195 g, 13.14 mmol) and Ph.sub.3P (3.83 g, 14.59 mmol) in
benzene (25 mL). Heat evolution was detected and the resulting
amber solution was stirred at ambient temperature for 6 h. Solvent
was removed under reduced pressure and the residue was purified via
BIOTAGE.RTM. (0 to 15% EtOAc/Hex; 80 g column). A white solid
corresponding to Cap-179, step b (0.77 g) was isolated. LC-MS:
Anal. Calcd. for [M].sup.+ C.sub.14H.sub.17NO.sub.5: 279.11; found
279.12. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.27-8.32
(2H, m), 8.20-8.24 (2H, m), 5.45 (1H, quin, J=2.82 Hz), 3.92 (2H,
dqd, J=11.90, 6.10, 6.10, 6.10, 1.53 Hz), 1.91 (2H, dd, J=14.80,
2.29 Hz), 1.57 (3H, dt, J=14.65, 3.05 Hz), 1.22 (6H, d, J=6.10
Hz).
Cap-179, Step c
##STR00252##
[0509] A solution LiOH (0.330 g, 13.8 mmol) in water (8 mL) was
added to a solution of Cap-179, step b (0.77 g, 2.76 mmol) in THF
(30 mL) and the resulting mixture was stirred at ambient
temperature for 16 h. THF was removed under reduced pressure and
the aqueous layer was diluted with more water (20 mL) and extracted
with EtOAc (3.times.15 mL). The combined organic layers were dried
(MgSO.sub.4), filtered and concentrated under vacuum. An oily
residue with a white solid was recovered. The mixture was
triturated with hexanes and the solid was filtered off to yield a
clear oil corresponding to Cap-179, step c (0.34 g). .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 4.21 (1H, quin, J=2.82 Hz),
3.87-3.95 (2H, m), 1.72 (1H, br. s.), 1.63 (2H, dd, J=14.34, 2.14
Hz), 1.39-1.47 (2H, m), 1.17 (6H, d, J=6.41 Hz).
Cap-179, Step d
##STR00253##
[0511] p-Tosyl chloride (3.98 g, 20.89 mmol) was added to a
solution of Cap-179, step c (1.36 g, 10.5 mmol) and Pyridine (3.38
mL, 41.8 mmol) in CH.sub.2Cl.sub.2 (150 mL) at room temperature and
stirred for 24 h and then concentrated to a yellow oil. The
remaining residue was added to pyridine (20 mL) and water (30 mL)
and the resulting mixture was stirred at ambient temperature for
11/2 h. The mixture was extracted with Et.sub.2O (75 mL) and the
separated organic layer was the washed thoroughly with 1 N aq. HCl
(4.times.50 mL). The organic layer was then dried (MgSO.sub.4),
filtered and concentrated. A white solid corresponding to Cap-179,
step d (2.2 g) was isolated. LC-MS: Anal. Calcd. for [2M+H].sup.+
C.sub.28H.sub.41O.sub.8S.sub.2: 569.22; found 569.3. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 7.80 (2H, d, J=8.28 Hz), 7.35
(2H, d, J=8.03 Hz), 4.89 (1H, quin, J=2.82 Hz), 3.77-3.88 (2H, m),
2.46 (3H, s), 1.77 (2H, dd, J=14.93, 2.89 Hz), 1.36 (2H, ddd,
J=14.31, 11.54, 2.76 Hz), 1.13 (6H, d, J=6.27 Hz).
Cap-179, Step e
##STR00254##
[0513] LiHMDS (4.30 mL, 4.30 mmol) was added to a solution of
Cap-179, step d (1.02 g, 3.59 mmol) and benzyl
2-(diphenylmethyleneamino)acetate (1.181 g, 3.59 mmol) in toluene
(25 mL) at room temperature in a sealed microwave vial and the
resulting mixture was then stirred for 5 h at 100.degree. C. under
microwave radiation. The reaction was quenched with water (10 mL),
extracted with EtOAc, washed with water, dried over MgSO.sub.4,
filtrated, and concentrated in vacuum. The residue was purified via
BIOTAGE.RTM. (0% to 6% EtOAc/Hex; 80 g column) and a yellow oil
corresponding to Cap-179, step e (1.2 g) was isolated. Anal. Calcd.
for [2M+Na].sup.+ C.sub.58H.sub.62N.sub.2NaO.sub.6: 905.45; found
905.42. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.64-7.70
(4H, m), 7.29-7.44 (29H, m), 7.06 (4H, dd, J=7.65, 1.63 Hz), 5.18
(2H, d, J=2.01 Hz), 3.89 (2H, d, J=6.53 Hz), 3.79-3.87 (1H, m),
3.46 (5H, dquind, J=11.26, 5.87, 5.87, 5.87, 5.87, 1.88 Hz), 2.47
(2H, s), 2.35-2.46 (2H, m), 1.78 (1H, dd, J=14.81, 3.01 Hz),
1.62-1.65 (1H, m), 1.61 (2H, s), 1.36-1.43 (3H, m), 1.19 (7H, d,
J=6.27 Hz), 1.14 (11H, dd, J=6.15, 2.89 Hz), 0.86-0.96 (3H, m).
Cap-179, step f (Enantiomer-1 and Enantiomer-2)
##STR00255##
[0515] Cap-179, step e (2.08 g, 4.71 mmol) was dissolved in THF
(100 mL) and treated with 2 N HCl (9.42 mL, 18.84 mmol). The
resulting clear solution was stirred at ambient temperature for 4 h
and then THF was removed under reduced pressure. The remaining
aqueous layer was extracted with hexanes (3.times.20 ml) and after
diluting with H.sub.2O (20 mL), the aqueous phase was basified with
1 N NaOH to pH=10 and extracted with EtOAc (3.times.10 mL). The
combined organic layers were dried (MgSO.sub.4), filtered and
concentrated under vacuum. The resulting residue was taken up in
CH.sub.2Cl.sub.2 (100 mL) and charged with DIEA (2.468 mL, 14.13
mmol) and methyl chloroformate (0.401 mL, 5.18 mmol). The resulting
solution was stirred at ambient temperature for 2 h. The reaction
mixture was quenched with water (10 mL) and the organic layer was
removed under reduced pressure. The aqueous layer was then
extracted with EtOAc (3.times.10 mL) and the combined organic
layers were dried (MgSO.sub.4), filtered and concentrated. The
residue was purified via BIOTAGE.RTM. (10% EtOAc/Hex; 25 g column).
A clear colorless oil corresponding to Cap-179, step f (1.05 g) was
recovered. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.18H.sub.26NO.sub.5: 336.18; found 336.3. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 7.32-7.40 (5H, m), 5.26 (1H, d, J=8.24
Hz), 5.13-5.24 (2H, m), 4.36 (1H, dd, J=8.85, 4.88 Hz), 3.68 (3H,
s), 3.32-3.46 (2H, m), 2.02-2.14 (1H, m), 1.52 (1H, d, J=12.82 Hz),
1.32 (1H, d, J=12.51 Hz), 1.11-1.18 (6H, m), 0.89-1.07 (2H, m).
[0516] A chiral SFC method was developed to separate the racemic
mixture by using 12% methanol as the modifier on a CHIRALPAK.RTM.
AD-H column (30.times.250 mm, 5 .mu.m) (Temp=35.degree. C.,
Pressure=150 bar, Wavelength=210 nm, Flow rate=70 mL/min for 8 min,
Solvent A=CO.sub.2, Solvent B=MeOH). The two separated isomers,
Cap-179 step f (Enantiomer-1) (first eluting) and Cap-179 step f
(Enantiomer-2) (second eluting) exhibited the same analytical data
as the corresponding mixture (see above).
Cap-179 (Enantiomer-1 and Enantiomer-2)
[0517] Cap-179 step f (Enantiomer-1) (0.35 g, 1.044 mmol) was
dissolved in MeOH (50 mL) in a Parr bottle and charged with Pd/C
(0.111 g, 1.044 mmol). The suspension was then placed in a Parr
shaker and the mixture was flushed with N.sub.2 (3.times.), placed
under 40 psi of H.sub.2 (2.104 mg, 1.044 mmol) and shaken at room
temperature for 2 h. The catalyst was filtered off through a pad of
CELITE.RTM. and the solvent was removed under reduced pressure, to
yield an amber solid corresponding to Cap-179 Enantiomer-1 (0.25
g). .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm 12.74 (4H, br.
s.), 7.35 (4H, d, J=6.10 Hz), 3.85 (4H, br. s.), 3.53 (3H, s), 3.35
(2H, ddd, J=15.95, 9.99, 6.10 Hz), 1.97 (1H, br. s.), 1.48 (2H, t,
J=13.28 Hz), 1.06 (6H, d, J=6.10 Hz), 0.82-1.00 (2H, m)
[0518] Cap-179 Enantiomer-2 was prepared similarly: .sup.1H NMR
(500 MHz, DMSO-d.sub.6) .delta. ppm 12.50 (1H, br. s.), 7.31 (1H,
br. s.), 3.84 (1H, t, J=7.32 Hz), 3.53 (3H, s), 3.29-3.41 (2H, m),
1.99 (1H, s), 1.48 (2H, t, J=14.34 Hz), 1.06 (6H, d, J=6.10 Hz),
0.95 (1H, q, J=12.21 Hz), 0.87 (1H, q, J=11.80 Hz). [Note: the
minor variation in the .sup.1H NMR profile of the enantiomers is
likely a result of a difference in sample concentration.]
Cap-180 (Racemic Mixture)
##STR00256##
[0519] Cap-180, Step a
##STR00257##
[0521] p-Tosyl-Cl (4.39 g, 23.0 mmol) was added to a solution of
Cap-179, step a (1.50 g, 11.5 mmol) and pyridine (3.73 mL, 46.1
mmol) in CH.sub.2Cl.sub.2 (50 mL) at room temperature and stirred
for 2 days. The reaction was diluted with CH.sub.2Cl.sub.2, washed
with water, then 1 N HCl. The organic layer was dried (MgSO.sub.4)
and concentrated to a yellow oil which was purified via
BIOTAGE.RTM. (5% to 20% EtOAc/Hex; 40 g column). A clear oil that
solidified under vacuum and corresponding to Cap-180, step a (2.89
g) was isolated. LC-MS: Anal. Calcd. for [2M+Na].sup.+
C.sub.28H.sub.40NaO.sub.8S.sub.2: 591.21; found 591.3. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 7.80 (2H, d, J=8.24 Hz), 7.35
(2H, d, J=7.93 Hz), 4.59 (1H, tt, J=11.37, 4.96 Hz), 3.36-3.46 (2H,
m), 2.46 (3H, s), 1.91 (2H, dd, J=12.05, 5.04 Hz), 1.37 (2H, dt,
J=12.67, 11.52 Hz), 1.19 (6H, d, J=6.10 Hz).
Cap-180, Step b
##STR00258##
[0523] LiHMDS 1 N (7.09 mL, 7.09 mmol) was added to a solution of
Cap-180, step a (1.68 g, 5.91 mmol) and ethyl
2-(diphenylmethyleneamino)acetate (1.579 g, 5.91 mmol) in toluene
(30 mL) at room temperature and the resulting mixture was then
stirred for 16 h at 85.degree. C. The reaction was quenched with
water (50 mL), extracted with EtOAc, washed with water, dried over
MgSO.sub.4, filtrated, and concentrated in vacuo. The residue was
purified via BIOTAGE.RTM. (0% to 15% EtOAc/Hex; 40 g column). A
clear yellowish oil corresponding to Cap-180, step b (racemic
mixture; 0.64 g) was isolated. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.24H.sub.30NO.sub.3: 380.22; found 380.03. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 7.64-7.70 (2H, m), 7.45-7.51 (3H, m),
7.38-7.44 (1H, m), 7.31-7.37 (2H, m), 7.13-7.19 (2H, m), 4.39 (1H,
d, J=10.54 Hz), 4.16-4.26 (2H, m), 3.29-3.39 (1H, m), 2.93-3.03
(1H, m), 2.70 (1H, m, J=9.41, 4.14 Hz), 1.42-1.49 (2H, m),
1.31-1.37 (1H, m), 1.29 (4H, t, J=7.15 Hz), 1.04 (6H, dd, J=7.78,
6.27 Hz).
Cap-180, Step c
##STR00259##
[0525] Cap-180, step b (0.36 g, 0.949 mmol) was dissolved in THF
(10 mL) and treated with 2 N HCl (1.897 mL, 3.79 mmol). The
resulting clear solution was stirred at ambient temperature for 20
h and THF was removed under reduced pressure. The remaining aqueous
layer was extracted with hexanes (3.times.20 mL) and after diluting
with H.sub.2O (20 mL), the aqueous phase was basified with 1 N NaOH
to pH=10 and extracted with EtOAc (3.times.10 mL). The combined
organic layers were dried (MgSO.sub.4), filtered and concentrated
under vacuum. The resulting residue was taken up in
CH.sub.2Cl.sub.2 (10.00 mL) and charged with DIEA (0.497 mL, 2.85
mmol) and methyl chloroformate (0.081 mL, 1.044 mmol). The
resulting solution was stirred at ambient temperature for 2 h and
the reaction mixture was quenched with water (10 mL) and the
organic layer was removed under reduced pressure. Aqueous layer was
extracted with EtOAc (3.times.10 mL) and the combined organic
layers were dried (MgSO.sub.4), filtered and concentrated. An amber
oil corresponding to Cap-180, step c (0.21 g) was recovered and it
was used without further purification. LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.13H.sub.24NO.sub.5: 273.17; found 274.06. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. ppm 5.20 (1H, d, J=8.03 Hz), 4.59
(1H, t, J=10.16 Hz), 4.11-4.27 (3H, m), 3.69-3.82 (2H, m), 3.64
(3H, s), 1.95-2.07 (1H, m), 1.63 (1H, d, J=13.80 Hz), 1.41 (2H, dd,
J=8.03, 4.02 Hz), 1.31-1.37 (1H, m), 1.26 (3H, t, J=7.15 Hz), 1.16
(1H, d, J=6.27 Hz), 1.12 (6H, dd, J=6.15, 3.89 Hz).
Cap-180 (Racemic Mixture)
[0526] Cap-180, step c (0.32 g, 1.2 mmol) was dissolved in THF (10
mL) and charged with LiOH (0.056 g, 2.342 mmol) in water (3.33 mL)
at 0.degree. C. The resulting solution was stirred at rt for 2 h.
THF was removed under reduced pressure and the remaining residue
was diluted with water (15 mL) and washed with Et.sub.2O
(2.times.10 mL). The aqueous layer was then acidified with 1N HCl
to pH .about.2 and extracted with EtOAc (3.times.15 mL). The
combined organic layers were dried (MgSO.sub.4), filtered and
concentrated under vacuum to yield Cap-180 (racemic mixture) (0.2
g) as a white foam. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.11H.sub.20NO.sub.5: 246.13; found 246.00. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm 5.14 (1H, d, J=9.03 Hz), 4.65 (1H, t,
J=9.91 Hz), 3.63-3.89 (5H, m), 1.99-2.13 (1H, m), 1.56-1.73 (2H,
m), 1.48-1.55 (1H, m), 1.35-1.48 (1H, m), 1.27 (1H, br. s.), 1.17
(6H, d, J=6.02 Hz).
Cap-181
##STR00260##
[0527] Cap-181, Step a
##STR00261##
[0529] A solution of tert-butyl diazoacetate (1.832 mL, 13.22 mmol)
in 50 mL of CH.sub.2Cl.sub.2 was added into a mixture of
2,5-dihydrofuran (9.76 mL, 132 mmol), Rhodium(II) acetate dimer
(0.058 g, 0.132 mmol) in 40 mL of CH.sub.2Cl.sub.2 dropwise by a
syringe pump over 5 hours. The resulting mixture was then stirred
at room temperature overnight. The solvent was removed under
vacuum. The residue was purified by chromatography (silica gel,
0%-15% EtOAc/Hex) to afford Cap-181 step a (trans-isomer) (720 mg)
and Cap-181, step a (cis-isomer) (360 mg) as clear oil. Cap-181
step a (trans-isomer): .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
ppm 3.88 (2H, d, J=8.55 Hz), 3.70 (2H, d, J=8.55 Hz), 2.03-2.07
(2H, m), 1.47 (1H, t, J=3.20 Hz), 1.41 (9H, s); Cap-1, step a
(cis-isomer): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 4.06
(2H, d, J=8.53 Hz), 3.73 (2H, d, J=8.03 Hz), 1.81-1.86 (2H, m),
1.65-1.71 (1H, m), 1.43-1.47 (9H, m).
Cap-181, Step b
##STR00262##
[0531] To a solution of (Cap-181, step a (trans-isomer)) (700 mg,
3.80 mmol) in 15 mL of diethyl ether at -10.degree. C. was added
LiAlH4 (7.60 mL, 7.60 mmol) (1 M in THF) dropwise over 1 hour. The
resulting mixture was stirred at -10.degree. C. for 1 hour then at
room temperature for 1 hour. The mixture was then cooled to
-5.degree. C. 10 mL of Rochelle's salt (potassium sodium tartrate)
aqueous solution was added dropwise to quench the reaction. The
mixture was stirred at room temperature for 30 min and then
extracted with EtOAc (3.times.). The combined organic layers were
dried with MgSO.sub.4 and concentrated to afford Cap-181, step b
(380 mg) as light yellow oil. The product was used in the next step
without purification. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
3.85 (2H, d, J=8.28 Hz), 3.68 (2 H, d, J=8.53 Hz), 3.45-3.55 (2H,
m), 1.50-1.56 (2H, m), 1.02-1.11 (1H, m).
Cap-181, Step c
##STR00263##
[0533] To a solution of DMSO (4.82 mL, 67.9 mmol) in
CH.sub.2Cl.sub.2 (70 mL) was added dropwise oxalyl chloride (3.14
mL, 35.8 mmol) at -78.degree. C. The resulting mixture was stirred
at -78.degree. C. for 15 min. A solution of Cap-181, step b (3.10
g, 27.2 mmol) in 35 mL of CH.sub.2Cl.sub.2 was added and the
mixture was stirred at -78.degree. C. for 1 hour. Et.sub.3N (18.93
mL, 136 mmol) was then added dropwise. After 30 min, the cooling
bath was removed and the reaction was quenched with cold 20%
K.sub.2HPO.sub.4 aq. solution (10 mL) and water. The mixture was
stirred at room temperature for 15 min and then diluted with
Et.sub.2O. The layers were separated. The aqueous layer was
extracted with Et.sub.2O (2.times.). The combined organic layers
were washed with brine, dried with MgSO.sub.4 and concentrated. The
residue was purified by flash chromatography (silica gel, 100%
CH.sub.2Cl.sub.2) to afford Cap-181, step c (2.71 g) as light
yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 9.41 (1H,
d, J=4.27 Hz), 3.96 (2H, d, J=8.85 Hz), 3.80 (2H, d, J=8.55 Hz),
2.27-2.33 (2H, m), 1.93 (1H, m).
Cap-181, Step d
##STR00264##
[0535] To a mixture of Cap-181, step c (2.7 g, 24.08 mmol) in 50 mL
of water at 0.degree. C. was added sodium bisulfate (2.506 g, 24.08
mmol) and KCN (1.631 g, 25.04 mmol), followed by a solution of
(R)-2-amino-2-phenylethanol (3.30 g, 24.08 mmol) in 18 mL of MeOH.
The resulting mixture was stirred at room temperature for 2 hours
and then heated to reflux overnight. The mixture was cooled to room
temperature. 100 mL of EtOAc was added. After mixing for 15 min,
the layers were separated. The aqueous layer was extracted with
EtOAc (2.times.). The combined organic layers were washed with
brine, dried with MgSO.sub.4 and concentrated. The crude
diastereomeric mixture was purified by reverse phase HPLC (Column:
Water Sunfire 30.times.150 mm, acetonitrile/water/NH.sub.4OAc) to
afford a two diastereomers of Cap-181, step d. The absolute
stereochemistry of each isomer was not determined Diastereomer 1
(later eluting fraction) (570 mg): LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.15H.sub.19N.sub.2O.sub.2 259.14; found 259.2.
Cap-181, Step e
##STR00265##
[0537] To a solution of Cap-181, step d (diastereomer 1) (570 mg,
2.207 mmol) in 20 mL of CH.sub.2Cl.sub.2 and 20 mL of MeOH at
0.degree. C. was added lead tetraacetate (1174 mg, 2.65 mmol). The
resulting orange mixture was stirred at 0.degree. C. for 10 min.
Water (20 mL) was then added into the mixture and the mixture was
filtered off (CELITE.RTM.). The filtrate was concentrated and
diluted with 25 mL of 6 N HCl aq. solution. The resulting mixture
was refluxed for 4 hours. The mixture was filtered off and washed
with CH.sub.2Cl.sub.2. The aqueous layer was concentrated to afford
Cap-181, step e (HCl salt). The crude product was used in the next
step without further purification. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 3.87-3.91 (2H, m), 3.73 (2H, dd, J=8.70, 2.90 Hz), 3.55
(1H, d, J=10.07 Hz), 2.02-2.07 (1 H, m), 1.94-1.99 (1H, m),
1.03-1.10 (1H, m).
Cap-181
[0538] To a mixture of the above crude Cap-181, step e in 1 N NaOH
aq. solution (10 mL) was added sodium bicarbonate (371 mg, 4.42
mmol). Methyl chloroformate (0.342 mL, 4.42 mmol) was then added
dropwise, and the resulting mixture was stirred at room temperature
for 3 hours. The mixture was neutralized with 1 N HCl aq. Solution
and extracted with EtOAc (3.times.). The combined organic layers
were dried with MgSO.sub.4 and concentrated to afford Cap-181 (100
mg, 21% over two steps) as light yellow oil. LC-MS: Anal. Calcd.
for [M+H].sup.+ C.sub.9H.sub.14NO.sub.5 216.09; found 216.1.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 5.29 (1H, br. s.),
3.53-4.02 (8H, m), 1.66-1.92 (2H, m), 1.08 (1H, br. s.).
Cap-182 (Racemic Mixture)
##STR00266##
[0539] Cap-182 Step a
##STR00267##
[0541] A solution of cyclopent-3-enol (5 g, 59.4 mmol) and
Et.sub.3N (9.94 mL, 71.3 mmol) in 50 mL of CH.sub.2Cl.sub.2 was
stirred at room temperature for 15 min. Benzoyl chloride (8.28 mL,
71.3 mmol) was then added dropwise and the mixture was stirred at
room temperature overnight. The mixture was then washed with water,
and the organic layer was dried with MgSO.sub.4 and concentrated.
The residue was purified by flash chromatography (silica gel,
EtOAc/Hex 0-10%) to afford Cap-182, step a (9.25 g) as clear oil.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.01-8.07 (2H, m),
7.55 (1H, t, J=7.40 Hz), 7.43 (2H, t, J=7.65 Hz), 5.79 (2H, s),
5.64 (1H, tt, J=6.93, 2.60 Hz), 2.87 (2H, dd, J=16.56, 6.78 Hz),
2.52-2.63 (2H, m).
Cap-182, Step b
##STR00268##
[0543] To a round bottom flask with a magnetic stirring bar was
added sodium fluoride (5.02 mg, 0.120 mmol) and Cap-182, step a
(2.25 g, 11.95 mmol). The flask was heated up to 100.degree. C. and
neat trimethylsilyl 2,2-difluoro-2-(fluorosulfonyl)acetate (5.89
mL, 29.9 mmol) was added slowly by syringe pump over 5 hours, and
heated at 100.degree. C. overnight. The mixture was then diluted
with CH.sub.2Cl.sub.2, washed with water, sat. NaHCO.sub.3 aq.
solution and brine, dried with MgSO.sub.4 and concentrated. The
crude product was purified by flash chromatography (silica gel,
0-5% EtOAc/Hex) to afford Cap-182, step b (isomer 1) (750 mg) and
Cap-182, step b (isomer 2) (480 mg) as clear oils. Relative
stereochemical assignment was made by NOE study. Cap-182, step b
(isomer 1): LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.13H.sub.13F.sub.2O.sub.2 239.09; found 239.2. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 7.99-8.04 (2H, m), 7.56 (1H, t,
J=7.32 Hz), 7.43 (2H, t, J=7.63 Hz), 5.25-5.33 (1 H, m), 2.50 (2H,
dd, J=14.04, 6.71 Hz), 2.14-2.22 (2H, m), 2.08-2.14 (2H, m).
Cap-182, step b (isomer 2): LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.13H.sub.13F.sub.2O.sub.2 239.09; found 239.2. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 7.98-8.08 (2H, m), 7.53-7.59 (1H,
m), 7.41-7.48 (2H, m), 5.53-5.62 (1H, m), 2.59-2.70 (2H, m),
2.01-2.11 (4H, m).
Cap-182, Step c
##STR00269##
[0545] To a solution of Cap-182, step b (isomer 2) (480 mg, 2.015
mmol) in 4 mL of MeOH was added KOH (4 mL, 2.015 mmol) (10% aq.).
The resulting mixture was stirred at room temperature overnight.
The mixture was then extracted with CH.sub.2Cl.sub.2 (3.times.).
The combined organic layers were dried with MgSO.sub.4 and
concentrated to afford Cap-182, step c (220 mg) as a light yellow
solid. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 4.41-4.54 (1
H, m), 2.38-2.50 (2H, m), 1.89-1.99 (2H, m), 1.81 (2H, dd, J=14.50,
5.04 Hz).
Cap-182, Step d
##STR00270##
[0547] p-Tosyl-Cl (625 mg, 3.28 mmol) was added to a solution of
Cap-182, step c (220 mg, 1.640 mmol) and pyridine (0.531 mL, 6.56
mmol) in 7 mL of CH.sub.2Cl.sub.2. The mixture was stirred at room
temperature overnight and then diluted with CH.sub.2Cl.sub.2,
washed with water and 1 N HCl aq. solution. The organic layer was
dried (MgSO.sub.4) and concentrated. The residue was purified by
flash chromatography (silica gel, 0-15% EtOAc/Hexane) to afford
Cap-182, step d (325 mg) as a clear oil. LC-MS: Anal. Calcd. For
[M+Na].sup.+ C.sub.13H.sub.14F.sub.2NaO.sub.3S 311.05; found 311.2.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 7.76 (2H, d, J=8.24
Hz), 7.34 (2H, d, J=8.24 Hz), 4.99-5.08 (1H, m), 2.45 (3H, s),
2.31-2.41 (2H, m), 1.84-1.94 (4H, m).
Cap-182, Step e (Racemic Mixture)
##STR00271##
[0549] To a microwave tube was added N-(diphenylmethylene)glycine
ethyl ester (241 mg, 0.902 mmol) and Cap-182, step d (260 mg, 0.902
mmol) in 2 mL of toluene. The tube was sealed and LiHMDS (1.082 mL
of 1 N in THF, 1.082 mmol) was added dropwise under N.sub.2. The
resulting dark brown solution was heated at 100.degree. C. in
microwave for 5 hours. The mixture was then quenched with water,
and extracted with EtOAc (3.times.). The combined organic layers
were washed with water, dried with MgSO.sub.4 and concentrated. The
crude product was purified by flash chromatography (silica gel,
0-5% EtOAc/Hex) to afford a racemic mixture of Cap-182, step e (240
mg) as light yellow oil. The mixture was submitted to the next step
without separation. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.23H.sub.24F.sub.2NO.sub.2 384.18, found 384.35. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 7.63-7.70 (2H, m), 7.43-7.51 (3H,
m), 7.38-7.43 (1H, m), 7.31-7.38 (2H, m), 7.13-7.22 (2H, m),
4.13-4.22 (2H, m), 3.95 (1H, d, J=6.41 Hz), 2.67-2.79 (1H, m),
2.07-2.16 (1H, m), 1.97-2.07 (2H, m), 1.90 (2H, m), 1.65-1.76 (1H,
m), 1.25 (3H, t, J=7.17 Hz).
Cap-182, Step f (Racemic Mixture)
##STR00272##
[0551] To a solution of Cap-182, step e (240 mg, 0.626 mmol) in 4
mL of THF was added HCl (1 mL, 2.0 mmol) (2 N aq.). The resulting
mixture was stirred at room temperature for 2 hours. The mixture
was then washed with EtOAc, neutralized with sat. NaHCO.sub.3 aq.
solution and then extracted with EtOAc (3.times.). The combined
organic layers were dried with MgSO.sub.4 and concentrated to
afford Cap-182, step f (120 mg) as clear oil. LC-MS: Anal. Calcd.
for [M+H].sup.+ C.sub.10H.sub.16F.sub.2NO.sub.2 220.11; found
220.26. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 4.14-4.25
(2H, m), 3.26 (1H, d, J=6.71 Hz), 2.22-2.35 (1H, m), 1.90-2.11 (5H,
m), 1.79-1.90 (1H, m), 1.22-1.34 (3H, m).
Cap-182, Step g (Racemic Mixture)
##STR00273##
[0553] To a solution of Cap-182, step f (120 mg, 0.547 mmol) in 2
mL of CH.sub.2Cl.sub.2 was added methyl chloroformate (0.085 mL,
1.095 mmol). The resulting mixture was stirred at room temperature
for 1 hour. The mixture was diluted with CH.sub.2Cl.sub.2 and
washed with water. The organic layer was dried with
Na.sub.2SO.sub.4 and concentrated to afford Cap-182, step g (150
mg) as a white solid. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.12H.sub.18F.sub.2NO.sub.4 278.12; found 278.2. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 5.23 (1H, d, J=8.24 Hz), 4.29
(1H, t, J=7.48 Hz), 4.15-4.23 (2H, m), 3.68 (3H, s), 2.37 (1H, br.
s.), 2.02-2.10 (1H, m), 1.85-2.00 (4H, m), 1.75-1.84 (1H, m), 1.27
(3H, t, J=7.02 Hz).
Cap-182 (Racemic Mixture)
[0554] To a mixture of Cap-182, step g (150 mg, 0.541 mmol) in 2 mL
of THF and 1 mL of water was added LiOH (0.811 mL, 1.623 mmol) (2 N
aq.). The resulting mixture was stirred at room temperature
overnight. The mixture was neutralized with 1 N HCl aq. solution
and extracted with EtOAc (3.times.). The combined organic layers
were dried with MgSO.sub.4 and concentrated to afford Cap-182 (133
mg) as a white solid. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.14F.sub.2NO.sub.4 250.09; found 250.13. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 5.18-5.36 (1H, m), 4.28-4.44 (1H,
m), 3.70 (3H, s), 2.37-2.56 (1H, m), 1.74-2.31 (6H, m).
Cap-183 (Racemic Mixture)
##STR00274##
[0556] Cap-183 was synthesized from Cap-182, step b (isomer 1)
according to the procedure described for the preparation of
Cap-182. Anal. Calcd. for [M+H].sup.+
C.sub.10H.sub.14F.sub.2NO.sub.4 250.09, found 249.86. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 5.15 (1H, d, J=8.24 Hz), 4.32
(1H, t, J=7.48 Hz), 3.69 (3H, s), 2.83-2.99 (1H, m), 1.96-2.26 (4H,
m), 1.70 (1H, t, J=11.75 Hz), 1.59 (1H, t, J=12.05 Hz).
Cap-184 (Racemic Mixture)
##STR00275##
[0557] Cap-184, Step a
##STR00276##
[0559] A mixture of ethyl
2-amino-2-((1R,3r,5S)-bicyclo[3.1.0]hexan-3-yl)acetate (prepared
from commercially available (1R,3r,5S)-bicyclo[3.1.0]hexan-3-ol by
employing the same procedures described for the preparation of
Cap-182; 350 mg, 1.910 mmol), DiPEA (0.667 mL, 3.82 mmol), methyl
chloroformate (0.296 mL, 3.82 mmol) in 5 mL of CH.sub.2Cl.sub.2 was
stirred at room temperature for 1 hour. The mixture was then
diluted with CH.sub.2Cl.sub.2 and washed with water. The organic
layer was dried with MgSO.sub.4 and concentrated to afford Cap-184
step a (461 mg) as yellow oil. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.12H.sub.20NO.sub.4 242.14; found 242.2. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 5.04 (1H, d, J=7.63 Hz), 4.09-4.20 (2H, m),
4.05 (1H, t, J=8.39 Hz), 3.63 (3H, s), 2.55-2.70 (1 H, m),
1.96-2.09 (2H, m), 1.37-1.60 (4H, m), 1.24 (3H, t, J=7.17 Hz),
0.66-0.76 (1 H, m), -0.03-0.06 (1H, m).
Cap-184 (Racemic Mixture)
[0560] To a mixture of ethyl ester Cap-184, step a (461 mg, 1.911
mmol) in 5 mL of THF and 2 mL of water was added LiOH (2.87 mL,
5.73 mmol) (2 N aq.). The resulting mixture was stirred at room
temperature overnight. The mixture was then neutralized with 1 N
HCl aqueous solution, and extracted with EtOAc (3.times.). The
combined organic layers were dried with MgSO.sub.4 and concentrated
to afford Cap-184 (350 mg) as clear oil. LC-MS: Anal. Calcd. for
[2M+Na].sup.+ C.sub.20H.sub.30N.sub.2NaO.sub.8 449.19; found 449.3.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 5.07 (1H, d, J=8.85
Hz), 4.13 (1H, t, J=8.24 Hz), 3.68 (3H, s), 2.64-2.79 (1H, m),
2.04-2.21 (2H, m), 1.23-1.49 (4H, m), 0.71-0.81 (1H, m), 0.03-0.12
(1H, m).
Cap-185 (Enantiomer-1 and Enantiomer-2)
##STR00277##
[0561] Cap-185, Step a
##STR00278##
[0563] To a mixture of furan (1.075 mL, 14.69 mmol) and zinc (1.585
g, 24.24 mmol) in 1 mL of THF was added
1,1,3,3-tetrabromopropan-2-one (8.23 g, 22.03 mmol) and triethyl
borate (5.25 mL, 30.8 mmol) in 4 mL of THF dropwise during 1 hour
in dark. The resulting mixture was stirred at room temperature in
dark for 17 hours. The resulting dark brown mixture was cooled to
-15.degree. C., and 6 mL of water was added. The mixture was warmed
to 0.degree. C. and stirred at this temperature for 30 min. The
mixture was then filtered and washed with ether. The filtrate was
diluted with water and extracted with ether (3.times.). The
combined organic layers were dried with MgSO.sub.4 and concentrated
to afford dark brown oil. The dark brown oil was dissolved in 6 mL
of MeOH and the solution was added dropwise to a mixture of zinc
(4.99 g, 76 mmol), copper(I) chloride (0.756 g, 7.64 mmol) and
ammonium chloride (5.4 g, 101 mmol) in 20 mL of MeOH. The reaction
temperature was maintained below 15.degree. C. during addition. The
mixture was then stirred at room temperature for 20 hours,
filtered, and the filtrate was diluted with water and extracted
with CH.sub.2Cl.sub.2 (3.times.). The combined organic layers were
dried with MgSO.sub.4 and concentrated. The crude product was
purified by flash chromatography (silica gel, 0-14% EtOAc/Hex) to
afford Cap-185, step a as a white solid (1.0 g) as a white solid,
which turned yellow soon. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
ppm 6.24 (2H, s), 5.01 (2H, d, J=4.88 Hz), 2.73 (2H, dd, J=16.94,
5.04 Hz), 2.31 (2H, d, J=16.79 Hz).
Cap-185, Step b
##STR00279##
[0565] To a solution of Cap-185, step a (240 mg, 1.933 mmol) in 2
mL of THF at -78.degree. C. was added L-selectride (3.87 mL, 3.87
mmol) (1 M in THF) dropwise over 100 min. The resulting mixture was
stirred at -78.degree. C. for 1 hour and then at room temperature
overnight. The mixture was then cooled to 0.degree. C., 4 mL of 20%
NaOH aqueous solution was added, followed by 2 mL of H.sub.2O.sub.2
(30% water solution) dropwise. The resulting mixture was stirred
for 1 hour and then neutralized with 6N HCl (.about.5 mL). The
aqueous layer was saturated with NaCl and extracted with
CH.sub.2Cl.sub.2 (3.times.). The combined organic layers were dried
with MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 0-40% EtOAc/Hex) to afford
Cap-185, step b (180 mg) as clear oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 6.49 (2H, s), 4.76 (2H, d, J=4.27 Hz), 3.99
(1H, t, J=5.77 Hz), 2.29 (2H, ddd, J=15.18, 5.65, 4.02 Hz),
1.70-1.78 (2H, m).
Cap-185, Step c
##STR00280##
[0567] p-Tosyl-Cl (544 mg, 2.85 mmol) was added to a solution of
Cap-185, step b (180 mg, 1.427 mmol) and pyridine (0.462 mL, 5.71
mmol) in 5 mL of CH.sub.2Cl.sub.2 (5 mL) and the mixture was
stirred at room temperature for 2 days. The reaction was diluted
with CH.sub.2Cl.sub.2 and washed with 1 N aq. HCl. The aqueous
layer was extracted with CH.sub.2Cl.sub.2 (2.times.). The combined
organic layers were dried with MgSO.sub.4 and concentrated. The
crude product was purified by flash chromatography (silica gel,
0-15% EtOAc/Hex) to afford Cap-185, step c (210 mg) as a white
solid. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 7.73 (2H, d,
J=8.24 Hz), 7.32 (2H, d, J=8.24 Hz), 6.25 (2H, s), 4.76 (1H, t,
J=5.65 Hz), 4.64 (2H, d, J=3.66 Hz), 2.44 (3H, s), 2.18 (2H, td,
J=10.07, 5.49 Hz), 1.71 (2H, d, J=15.56 Hz).
Cap-185, Step d
##STR00281##
[0569] A microwave tube was charged with benzyl
2-(diphenylmethyleneamino)acetate (1.5 g, 4.57 mmol) and Cap-185,
step c (1.28 g, 4.57 mmol) in 5 mL of toluene. The tube was sealed
and LiHMDS (5.5 mL, 5.5 mmol) (1 N in toluene) was added dropwise
under N.sub.2. The resulting dark brown solution was heated at
100.degree. C. in microwave for 5 hours. To the mixture was then
added water and EtOAc. The layers were separated and the water
phase was extracted with EtOAc (2.times.). The combined organic
layers were concentrated to afford Cap-185, step d as a racemic
mixture of. The crude mixture was submitted to the next step
without purification or separation. LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.29H.sub.28NO.sub.3 438.21; found 438.4.
Cap-185, Step e
##STR00282##
[0571] To a solution of the racemic mixture of Cap-185, step d in
30 mL of THF was added HCl (20 mL) (2 N aq.). The resulting mixture
was stirred at room temperature for 2 hours. After the reaction was
done as judged by TLC, the two layers were separated. The aqueous
layer was washed with EtOAc, neutralized with sat. NaHCO.sub.3 aq.
solution and then extracted with EtOAc (3.times.). The combined
organic layers were dried with MgSO.sub.4 and concentrated to
afford Cap-185, step e. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.16H.sub.20NO.sub.3 274.14; found 274.12.
Cap-185, Step f
##STR00283##
[0573] A solution of the crude Cap-185, step e, DiPEA (1.24 mL, 7.1
mmol) and methyl chloroformate (0.55 mL, 7.1 mmol) in 5 mL of
CH.sub.2Cl.sub.2 was stirred at room temperature for 1 hour. The
mixture was then diluted with CH.sub.2Cl.sub.2 and washed with
water. The organic layer was dried with Na.sub.2SO.sub.4 and
concentrated. The crude product was purified by flash
chromatography (silica gel, 0-40% EtOAc/Hex) to afford 700 mg of
the racemic mixture. The mixture was then separated by chiral HPLC
(CHIRALPAK.RTM. AD-H column, 30.times.250 mm, 5 um) eluting with
88% CO.sub.2-12% EtOH at 70 mL/min to afford 240 mg of Enantiomer-1
and 310 mg of Enantiomer-2 of Cap-1, step f as white solids.
Enantiomer-1: LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.18H.sub.22NO.sub.5 332.15; found 332.3. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 7.30-7.40 (5H, m), 6.03-6.16 (2H, m),
5.09-5.26 (3H, m), 4.65-4.74 (2H, m), 4.33 (1H, dd, J=9.16, 4.88
Hz), 3.67 (3H, s), 2.27-2.38 (1 H, m), 1.61-1.69 (1H, m), 1.45-1.56
(1H, m), 1.34 (1H, dd, J=13.43, 5.19 Hz), 1.07 (1H, dd, J=13.12,
5.19 Hz). Enantiomer-2: LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.18H.sub.22NO.sub.5 332.15; found 332.06.
Cap-185 (Enantiomer-1 and Enantiomer-2)
[0574] To a hydrogenation bottle containing a solution Cap-185,
step f (Enantiomer-2) (300 mg, 0.905 mmol) in 10 mL of MeOH was
added Pd/C (15 mg, 0.141 mmol) under a cover of nitrogen. The
mixture was hydrogenated on a Parr shaker at 40 psi for 3
hours.
[0575] The mixture was then filtered and the filtrate was
concentrated to afford Cap-185 (Enantiomer-2) (200 mg) as a white
solid. LC-MS: Anal. Calcd. for [M+H].sup.+ C.sub.11H.sub.18NO.sub.5
244.12; found 244.2. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
5.33 (1H, br. s.), 4.46 (2H, d), 4.28 (1H, br. s.), 3.68 (3H, s),
2.35 (1H, br. s.), 1.91-2.03 (2H, m), 1.56-1.80 (4H, m), 1.36-1.55
(2H, m). [Note: Cap-185 (Enantiomer-1) can be obtained in a similar
fashion.]
Cap-186
##STR00284##
[0577] To a solution of the ester Cap-185, step f (Enantiomer-2)
(150 mg, 0.453 mmol) in 4 mL of MeOH was added NaOH (4 mL of 1 N in
water, 4.00 mmol). The resulting mixture was stirred at room
temperature for 3 hours. The methanol was then removed under
vacuum, and the residue was neutralized with 1 N HCl solution and
extracted with EtOAc (3.times.). The combined organic layers were
dried with MgSO.sub.4 and concentrated to afford Cap-186 that was
contaminated with some benzyl alcohol (sticky white solid; 115 mg).
LC-MS: Anal. Calcd. for [M+H].sup.+ C.sub.11H.sub.16NO.sub.5
242.10; found 242.1. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
6.10-6.19 (2H, m), 5.36 (1H, d, J=8.85 Hz), 4.75-4.84 (2H, m), 4.28
(1H, dd, J=8.55, 4.58 Hz), 3.68 (3H, s), 2.33-2.45 (1H, m),
1.60-1.72 (2H, m), 1.30-1.48 (2H, m).
Cap-187
##STR00285##
[0578] Cap-187, Step a
##STR00286##
[0580] To a solution of Cap-178, step e (2.2 g, 18.94 mmol),
PPh.sub.3 (24.84 g, 95 mmol) and 4-nitrobenzoic acid (14.24 g, 85
mmol) in 30 mL of benzene was added DEAD (42.9 mL, 95 mmol)
dropwise. The resulting light orange solution was stirred at room
temperature overnight. The solvent was then removed under vacuum
and the residue was purified by flash chromatography (silica gel,
0-15% EtOAc/Hex) to afford Cap-187, step a (2.3 g) as a white
solid. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 8.27-8.34 (2H,
m), 8.20-8.26 (2H, m), 5.45 (1H, t, J=2.90 Hz), 3.83-3.96 (3H, m),
1.90-2.03 (2H, m), 1.80-1.88 (1H, m), 1.61-1.70 (1H, m), 1.21 (3H,
d, J=6.10 Hz).
Cap-187, Step b
##STR00287##
[0582] To a solution of Cap-187, step a (2.3 g, 8.67 mmol) in 10 mL
of MeOH was added sodium methoxide (2.372 mL, 8.67 mmol) (25% in
Methanol). The resulting mixture was stirred at room temperature
for 3 hours. Water was added, and the mixture was extracted with
EtOAc (5.times.). The combined organic layers were dried with
MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 0-15% EtOAc/Hex, then 15-50%
EtOAc/Hex) to afford Cap-187, step b (0.85 g) as clear oil. .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. ppm 4.19-4.23 (1H, m), 3.82-3.91
(2H, m), 3.73-3.79 (1H, m), 1.79-1.88 (1H, m), 1.62-1.68 (1H, m),
1.46-1.58 (2H, m), 1.14 (3H, d, J=6.10 Hz).
Cap-187
[0583] The individual enantiomers of Cap-187 were synthesized from
Cap-187, step b according to the procedure described for Cap-178.
LC-MS: Anal. Calcd. for [M+H].sup.+ C.sub.10H.sub.18NO.sub.5
232.12; found 232.1. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
5.26 (1H, d, J=7.78 Hz), 4.32-4.43 (1H, m), 4.07 (1H, dd, J=11.54,
3.51 Hz), 3.72 (3H, s), 3.39-3.50 (2H, m), 2.08-2.23 (1H, m),
1.54-1.68 (1H, m), 1.38-1.52 (1H, m), 1.11-1.32 (5H, m).
Cap-188 (Four Stereoisomers)
##STR00288##
[0584] Cap-188, Step a
##STR00289##
[0586] To a solution of 2,2-dimethyldihydro-2H-pyran-4(3H)-one (2
g, 15.60 mmol) in 50 ml, of MeOH was slowly added sodium
borohydride (0.649 g, 17.16 mmol). The resulting mixture was
stirred at room temperature for 3 hours. To the mixture was then
added 1 N HCl aqueous solution until it crosses into acidic pH
range and then extracted with EtOAc (3.times.). The combined
organic layers were dried with MgSO.sub.4 and concentrated to
afford Cap-188, step a (1.9 g) as clear oil. The product was used
in the next step without purification. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 3.91-4.02 (1H, m), 3.79-3.86 (1H, m), 3.63
(1H, td, J=12.05, 2.51 Hz), 1.82-1.93 (2H, m), 1.40-1.53 (1H, m),
1.29-1.38 (1H, m), 1.27 (3H, s), 1.20 (3H, s).
Cap-188.1 and Cap-188.2, Step b
##STR00290##
[0588] p-Tosyl-Cl (5.56 g, 29.2 mmol) was added to a solution of
Cap-188, step a (1.9 g, 14.59 mmol) and pyridine (4.72 mL, 58.4
mmol) in 100 mL of CH.sub.2Cl.sub.2. The resulting mixture was
stirred at room temperature for 3 days. To the reaction was added
10 mL of water, and the mixture was stirred at room temperature for
an additional hour. The two layers were separated and the organic
phase was washed with water and 1 NHCl aqueous solution. The
organic phase was dried with MgSO.sub.4 and concentrated to afford
the mixture of two enantiomers as a light yellow solid. The mixture
was then separated by chiral HPLC (CHIRALPAK.RTM. AD column,
21.times.250 mm, 10 um) eluting with 92% 0.1%
diethylamine/Heptane-8% EtOH at 15 mL/min to afford Cap-188.1, step
b (1.0 g) and Cap-188.2, step b (1.0 g). The absolute
stereochemistry of the two enantiomers was not assigned. Cap-188.1,
step b: LC-MS: Anal. Calcd. for [2M+Na].sup.+
C.sub.28H.sub.40NaO.sub.8S.sub.2 591.21; found 591.3. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 7.79 (2H, d, J=8.24 Hz), 7.34
(2H, d, J=8.24 Hz), 4.72-4.81 (1H, m), 3.78 (1H, dt, J=12.44, 4.16
Hz), 3.53-3.61 (1 H, m), 2.45 (3H, s), 1.75-1.86 (2H, m), 1.61-1.71
(1H, m), 1.52-1.60 (1H, m), 1.22 (3H, s), 1.14 (3H, s). Cap-188.2,
step b: LC-MS: Anal. Calcd. for [2M+Na].sup.+
C.sub.28H.sub.40NaO.sub.8S.sub.2 591.21; found 591.3;
Cap-188
[0589] The four stereoisomers of Cap-188 could be synthesized from
Cap-188.1, step b and Cap-188.2, step b, according to the procedure
described for the preparation of Cap-178. Cap-188 (Steroisomer-1):
LC-MS: Anal. Calcd. for [M+Na].sup.+ C.sub.11H.sub.19NNaO.sub.5
268.12; found 268.23. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
5.32 (1H, d, J=8.55 Hz), 4.26-4.35 (1H, m), 3.57-3.82 (5H, m),
2.11-2.34 (1H, m), 1.25-1.58 (4H, m), 1.21 (6H, d, J=6.10 Hz).
Cap-188 (Stereoisomer-2): LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.11H.sub.20NO.sub.5 246.13; found 246.1. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 5.25 (1H, d, J=8.55 Hz), 4.33 (1H, dd,
J=8.39, 5.04 Hz), 3.80 (1H, dd, J=11.90, 3.97 Hz), 3.62-3.76 (4H,
m), 2.20-2.32 (1H, m), 1.52-1.63 (1H, m), 1.27-1.49 (3H, m), 1.22
(6H, d, J=14.04 Hz).
Cap-189
##STR00291##
[0590] Cap-189, Step a
##STR00292##
[0592] To a solution of phenylmagnesium bromide (113 mL, 340 mmol)
(3 M in ether) in 100 mL of ether was added dropwise
exo-2,3-epoxynorbornane (25 g, 227 mmol) in 50 mL of ether. After
the initial exotherm, the mixture was heated to reflux overnight.
The reaction was then cooled to room temperature and quenched
carefully with water (.about.10 mL). The mixture was diluted with
ether and washed with a 3 N HCl aqueous solution (.about.160 mL).
The aqueous layer was extracted with ether (2.times.) and the
combined organic layers were dried with MgSO.sub.4 and
concentrated. The crude product was purified by flash
chromatography (silica gel, 0-18% EtOAc/Hex) to afford Cap-189,
step a (11 g). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
6.03-6.11 (2H, m), 3.76 (1H, d, J=11.29 Hz), 2.72-2.81 (2H, m),
1.98 (1H, d, J=11.29 Hz), 1.67-1.76 (2H, m), 0.90-0.97 (2H, m).
Cap-189, step b
##STR00293##
[0594] To a solution of oxalyl chloride (59.9 mL, 120 mmol) in 200
mL of CH.sub.2Cl.sub.2 at -78.degree. C. was added DMSO (17.01 mL,
240 mmol) in 100 mL of CH.sub.2Cl.sub.2. The mixture was stirred
for 10 min, and Cap-189, step a (11 g, 100 mmol) in 150 mL of
CH.sub.2Cl.sub.2 was added followed by Et.sub.3N (72.4 mL, 519
mmol) in 30 mL of CH.sub.2Cl.sub.2. The mixture was stirred at
-78.degree. C. for 30 min and then warmed to room temperature.
Water (150 mL) was added and the mixture was stirred at room
temperature for 30 mins. The two layers were then separated, and
the aqueous layer was extracted with CH.sub.2Cl.sub.2 (2.times.).
The organic layers were combined, dried with MgSO.sub.4 and
concentrated. The crude product was purified by flash
chromatography (silica gel, 0-5% EtOAc/Hex) to afford Cap-189, step
b (5.3 g) as a light yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. ppm 6.50-6.55 (2H, m), 2.78-2.84 (2 H, m), 1.92-1.99 (2H,
m), 1.17-1.23 (2H, m).
Cap-189, step c
##STR00294##
[0596] A mixture of Cap-189, step b (5.3 g, 49.0 mmol),
p-toluenesulfonic acid monohydrate (1.492 g, 7.84 mmol) and
ethylene glycol (4.10 mL, 73.5 mmol) in 100 mL of benzene was
refluxed for 4 hours and then stirred at room temperature
overnight. The reaction was partitioned between Et.sub.2O and
aqueous sat. NaHCO.sub.3 solution and the two layers were
separated. The organic layer was washed with brine, dried with
MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 0-6% EtOAc/Hex) to afford
Cap-189, step c (5.2 g) as a clear oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 6.20 (2H, t, J=2.13 Hz), 3.90-3.97 (2H, m),
3.81-3.89 (2H, m), 2.54 (2H, m), 1.89-1.99 (2H, m), 0.95-1.03 (2H,
m).
Cap-189, Step d
##STR00295##
[0598] A solution of Cap-189, step c (5.2 g, 34.2 mmol) in 60 mL of
MeOH and 50 mL of CH.sub.2Cl.sub.2 was cooled to -78.degree. C. and
treated with ozone gas until a light blue color was apparent. The
reaction was then bubbled with N.sub.2 to remove the excess ozone
gas (blue color disappeared) and sodium borohydride (1.939 g, 51.3
mmol) was added into the reaction. The reaction was then warmed to
0.degree. C. Acetone was added into the mixture to quench the
excess sodium borohydride. The mixture was concentrated and the
residue was purified by flash chromatography (silica gel, 100%
EtOAc) to afford Cap-189, step d (5.0 g) as a clear oil. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.99-4.09 (4H, m), 3.68 (4H,
m), 2.17-2.29 (2H, m), 1.92-2.10 (2H, m), 1.77-1.88 (2H, m),
1.57-1.70 (2H, m).
Cap-189, Step e
##STR00296##
[0600] To a solution of Cap-189, step d (1 g, 5.31 mmol) in 20 mL
of CH.sub.2Cl.sub.2 was added silver oxide (3.8 g), p-Ts-Cl (1.215
g, 6.38 mmol) and KI (0.176 g, 1.063 mmol). The resulting solution
was stirred at room temperature for 3 days. The mixture was then
filtered and the filtrate was concentrated. The crude product was
purified by flash chromatography (silica gel, 60% EtOAc/Hex) to
afford Cap-189, step e (0.79 g) as clear oil. LC-MS: Anal. Calcd.
for [M+Na].sup.+ C.sub.16H.sub.22NaO.sub.6S 365.10; found 365.22.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.80 (2H, d, J=8.28
Hz), 7.36 (2H, d, J=8.03 Hz), 4.11-4.17 (1H, m), 3.85-4.06 (5H, m),
3.64-3.71 (1H, m), 3.55-3.63 (1H, m), 2.47 (3H, s), 2.32-2.43 (1H,
m), 2.15-2.27 (1H, m), 1.70-1.89 (2H, m), 1.52-1.66 (1H, m),
1.35-1.47 (1H, m).
Cap-189, Step f
##STR00297##
[0602] To a solution of Cap-189, step e (2.2 g, 6.43 mmol) in 40 mL
of MeOH was added potassium carbonate (1.776 g, 12.85 mmol). The
resulting mixture was stirred at room temperature overnight. The
mixture was then diluted with water and EtOAc. The two layers were
separated. The aqueous layer was extracted with EtOAc (2.times.).
The combined organic layers were washed with brine, dried with
MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 0-15% EtOAc/Hex) to afford
Cap-189, step f (0.89 g, 5.23 mmol, 81%) as clear oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 3.89-4.02 (6H, m), 3.58 (2H, dd,
J=10.79, 2.51 Hz), 1.69-1.89 (6H, m).
Cap-189, Step g
##STR00298##
[0604] To the solution of Cap-189, step f (890 mg, 5.23 mmol) in 15
mL of THF was added HCl (15 mL, 45.0 mmol) (3 M aqueous). The
resulting mixture was stirred at room temperature overnight. The
mixture was then diluted with ether and the two layers were
separated. The aqueous phase was extracted with ether (2.times.)
and the combined organic layers were dried with MgSO.sub.4 and
concentrated to afford Cap-189, step g (0.95 g, containing some
residual solvents). The product was used in the next step without
purification. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
3.95-4.00 (2H, m), 3.85 (2H, d, J=10.68 Hz), 2.21-2.28 (2H, m),
1.99-2.04 (2H, m), 1.90-1.96 (2H, m).
Cap-189, step h (Enantiomer-1 and Enantiomer-2)
##STR00299##
[0606] To a solution of
(+/-)-benzyloxycarbonyl-.alpha.-phosphonoglycine trimethyl ester
(1733 mg, 5.23 mmol) in 6 mL of THF at -20.degree. C. was added
1,1,3,3-tetramethylguanidine (0.723 mL, 5.75 mmol). The resultant
light yellow mixture was stirred at -20.degree. C. for 1 hour, and
Cap-189, step g (660 mg, 5.23 mmol) in 3 mL of THF was added and
mixture was then stirred at room temperature for 3 days. The
reaction mixture was then diluted with EtOAc, washed with a 0.1 N
HCl aq. solution. The aqueous layer was extracted with EtOAc
(2.times.) and the combined organic layers were dried with
MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 0-4% EtOAc/CH.sub.2Cl.sub.2) to
afford 960 mg of the racemic mixture. The mixture was separated by
chiral HPLC (CHIRALPAK.RTM. AD column, 21.times.250 mm, 10 um)
eluting with 90% 0.1% diethylamine/Heptane-10% EtOH at 15 mL/min to
afford Cap-189, step h (Enantiomer-1; 300 mg) and Cap-189, step h
(Enantiomer-2; 310 mg) as white solids. Cap-189, step h
(Enantiomer-1): LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.18H.sub.22NO.sub.5 332.15; found 332.2. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 7.29-7.41 (5H, m), 6.00 (1H, br. s.), 5.13
(2H, s), 3.63-3.87 (8H, m), 2.84 (1H, br. s.), 1.84-2.02 (2H, m),
1.63-1.84 (2H, m). Cap-189, step h (Enantiomer-2): LC-MS: Anal.
Calcd. for [M+H].sup.+ C.sub.18H.sub.22NO.sub.5 332.15; found
332.2.
Cap-189, step i
##STR00300##
[0608] N.sub.2 was bubbled through a solution of Cap-189, step h
(Enantiomer-2; 290 mg, 0.875 mmol) in 10 mL of MeOH in a 500 mL
hydrogenation bottle for 30 mins. To the solution was added
(S,S)-Me-BPE-Rh (9.74 mg, 0.018 mmol), and the mixture was then
hydrogenated at 60 psi for 6 days. The mixture was concentrated,
and chiral analytical HPLC (CHIRALPAK.RTM. OJ column) indicated
that there were a small amount of remaining starting material and
one major product. The residue was then separated by chiral HPLC
(CHIRALPAK.RTM. OJ column, 21.times.250 mm, 10 um) eluting with 70%
0.1% diethylamine/Heptane-30% EtOH at 15 mL/min to afford Cap-189,
step i, (150 mg) as clear oil. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.18H.sub.24NO.sub.5 334.17; found 334.39. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 7.28-7.41 (5H, m), 5.12-5.18 (1H, m),
5.09 (2H, s), 4.05 (1 H, t, J=10.07 Hz), 3.75 (3H, s), 3.60-3.72
(2H, m), 3.41-3.50 (2H, m), 2.10 (1H, br. s.), 1.72-1.99 (6H,
m).
Cap-189, Step j
##STR00301##
[0610] To a solution of Cap-189, step i (150 mg, 0.450 mmol) in 10
mL of MeOH in a hydrogenation bottle were added dimethyl
dicarbonate (0.072 mL, 0.675 mmol) and 10% Pd/C (23.94 mg, 0.022
mmol) under a cover of nitrogen cover. The mixture was then
hydrogenated on Parr-shaker at 45 psi overnight. The mixture was
filtered and the filtrate was concentrated to afford Cap-189, step
j (110 mg) as a clear oil. LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.12H.sub.20NO.sub.5 258.13; found 258.19. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 5.08 (1 H, d, J=9.16 Hz), 4.03 (1H, t,
J=10.07 Hz), 3.75 (3H, s), 3.60-3.72 (5H, m), 3.46 (2H, t, J=10.38
Hz), 2.11 (1H, br. s.), 1.72-1.99 (6H, m).
Cap-189
[0611] To a mixture of Cap-189, step j (110 mg, 0.428 mmol) in 2 mL
of THF and 1 mL of water was added LiOH (0.641 mL, 1.283 mmol) (2 N
aq.). The resulting mixture was stirred at room temperature
overnight. The mixture was neutralized with a 1 N HCl aq. solution
and extracted with EtOAc (3.times.). The combined organic layers
were dried with MgSO.sub.4 and concentrated to afford Cap-189 (100
mg) as a white solid. LC-MS: Anal. Calcd. for [M+Na].sup.+
C.sub.11H.sub.17NNaO.sub.5 266.10; found 266.21. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 5.10 (1H, d, J=9.16 Hz), 4.02 (1H, t,
J=10.07 Hz), 3.62-3.78 (5H, m), 3.49 (2H, d, J=10.68 Hz), 2.07-2.22
(2H, m), 1.72-1.98 (6H, m).
Cap-190 (Diastereomeric Mixture)
##STR00302##
[0612] Cap-190, Step a
##STR00303##
[0614] To a mixture of cyclopent-3-enol (2.93 g, 34.8 mmol) and
imidazole (5.22 g, 77 mmol) in 30 mL of DMF at 0.degree. C. was
added t-butyldimethylchlorosilane (6.30 g, 41.8 mmol). The
resulting colorless mixture was stirred at room temperature
overnight. Hexanes and water were then added to the mixture and the
two layers were separated. The aqueous layer was extracted with
EtOAc (2.times.) and the combined organic layers were washed with
brine, dried with MgSO.sub.4 and concentrated. The crude product
was purified by flash chromatography (silica gel, 2% EtOAc/Hex) to
afford Cap-190, step a (6.3 g) as a clear oil. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm 5.65 (2H, s), 4.49-4.56 (1H, m), 2.56
(2 H, dd, J=15.26, 7.02 Hz), 2.27 (2H, dd, J=15.26, 3.36 Hz), 0.88
(9H, s), 0.06 (6H, s).
Cap-190, Step b
##STR00304##
[0616] To a solution of Cap-190, step a (2.3 g, 11.59 mmol) in 40
mL of CH.sub.2Cl.sub.2 at 0.degree. C. was added m-CPBA (5.60 g,
16.23 mmol) in 5 portions. The reaction mixture was stirred at room
temperature overnight. Hexanes and water were then added to the
mixture and the two layers were separated. The organic layer was
washed with 50 mL aq. 10% NaHSO.sub.3 and brine, dried with
MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 3%-6% EtOAc/Hex) to afford
Cap-190, step b (1.42 g) and its trans diastereomer (0.53 g) as
clear oils. Cap-190, step b (cis): .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 4.39-4.47 (1H, m), 3.47 (2H, s), 2.01-2.10
(2H, m), 1.93-2.00 (2H, m), 0.88 (9H, s), 0.04 (6H, s). Cap-190,
step b (trans): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
4.04-4.14 (1H, m), 3.47 (2H, s), 2.41 (2H, dd, J=14.05, 7.28 Hz),
1.61 (2H, dd, J=14.18, 6.90 Hz), 0.87 (9H, s), 0.03 (6H, s).
Cap-190, step c
##STR00305##
[0618] To a solution of (S)-1,2'-methylenedipyrrolidine (0.831 g,
5.39 mmol) in 15 mL of benzene at 0.degree. C. was added dropwise
n-butyllithium (4.90 mL, 4.90 mmol) (1 M in hexane). The solution
turned bright yellow. The mixture was stirred at 0.degree. C. for
30 min. Cap-190, step b (cis-isomer; 0.7 g, 3.27 mmol) in 10 mL of
benzene was then added and the resulting mixture was stirred at
0.degree. C. for 3 hours. EtOAc and sat. NH.sub.4Cl aq. solution
were added into the mixture, and the two layers were separated. The
organic layer was washed with water and brine, dried with
MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 15% EtOAc/Hex) to afford Cap-190,
step c (400 mg) as a light yellow oil. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 5.84-5.98 (2H, m), 4.53-4.69 (2H, m),
2.63-2.73 (1H, m), 1.51 (1H, dt, J=13.73, 4.43 Hz), 0.89 (9H, s),
0.08 (6H, s).
Cap-190, Step d
##STR00306##
[0620] To a solution of Cap-190, step c (400 mg, 1.866 mmol), MeI
(1.866 mL, 3.73 mmol) (2 M in t-butyl methyl ether) in 5 mL of THF
at 0.degree. C. was added NaH (112 mg, 2.80 mmol) (60% in mineral
oil). The resulting mixture was allowed to warm up to room
temperature and stirred at room temperature overnight. The reaction
was then quenched with water and extracted with EtOAc (3.times.).
The combined organic layers were washed with brine, dried with
MgSO.sub.4 and concentrated. The crude product was purified by
flash chromatography (silica gel, 5% EtOAc/Hex) to afford Cap-190,
step d (370 mg) as light yellow oil. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 5.92-5.96 (1H, m), 5.87-5.91 (1H, m),
4.64-4.69 (1H, m), 4.23-4.28 (1H, m), 3.32 (3H, s), 2.62-2.69 (1H,
m), 1.54 (1H, dt, J=13.12, 5.49 Hz), 0.89 (9H, s), 0.07 (5H, d,
J=1.83 Hz).
Cap-190, Step e
##STR00307##
[0622] To a solution of Cap-190, step d (400 mg, 1.751 mmol) in 10
mL of EtOAc in a hydrogenation bottle was added platinum(IV) oxide
(50 mg, 0.220 mmol). The resulting mixture was hydrogenated at 50
psi on Parr shaker for 2 hours. The mixture was then filtered
through CELITE.RTM., and the filtrate was concentrated to afford
Cap-190, step e (400 mg) as a clear oil. LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.12H.sub.27O.sub.2Si 231.18; found 231.3. .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. ppm 4.10-4.17 (1H, m), 3.65-3.74
(1H, m), 3.27 (3H, s), 1.43-1.80 (6H, m), 0.90 (9H, s), 0.09 (6H,
s).
Cap-190, Step f
##STR00308##
[0624] To a solution of Cap-190, step e (400 mg, 1.736 mmol) in 5
mL of THF was added TBAF (3.65 mL, 3.65 mmol) (1 N in THF). The
color of the mixture turned brown after several min., and it was
stirred at room temperature overnight. The volatile component was
removed under vacuum, and the residue was purified by flash
chromatography (silica gel, 0-25% EtOAc/Hex) to afford Cap-190,
step f (105 mg) as light yellow oil. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 4.25 (1H, br. s.), 3.84-3.92 (1H, m), 3.29
(3H, s), 1.67-2.02 (6H, m).
Cap-190
[0625] Cap-190 was then synthesized from Cap-190, step f according
to the procedure described for Cap-182. LC-MS: Anal. Calcd. for
[M+Na].sup.+ C.sub.10H.sub.17NNaO.sub.5 254.10; found 254.3.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 5.25 (1H, d, J=8.55
Hz), 4.27-4.41 (1H, m), 3.81-3.90 (1H, m), 3.69 (3H, s), 3.26 (3H,
s), 2.46-2.58 (1H, m), 1.76-1.99 (3H, m), 1.64-1.73 (1H, m),
1.40-1.58 (1H, m), 1.22-1.38 (1H, m).
Cap-191 (Enantiomer-1)
##STR00309##
[0626] Cap-191, Step a
##STR00310##
[0628] To a solution of diisopropylamine (3 ml, 21.05 mmol) in THF
(3 ml) at -78.degree. C. under nitrogen was added n-butyl lithium
(2.5 M in hexanes; 8.5 ml, 21.25 mmol). The reaction was stirred at
-78.degree. C. for 10 min then brought up to 0.degree. C. for 25
min. The reaction was cooled down again to -78.degree. C., methyl
tetrahydro-2H-pyran-4-carboxylate (3 g, 20.81 mmol) in THF (3 ml)
was added. The reaction was stirred at -78.degree. C. for 15 min
then brought up to 0.degree. C. for 30 min. The reaction was cooled
down to -78.degree. C., methyl iodide (1.301 ml, 20.81 mmol) was
added. After the addition, the cold bath was removed and the
reaction was allowed to slowly warm up to .about.25.degree. C. and
stirred for 22 h. Ethyl acetate and aqueous HCl (0.1N) were added,
and the organic layer was separated and washed with brine and dried
(MgSO.sub.4), filtered, and concentrated in vacuo. The residue was
loaded on a Thomson's silica gel cartridge eluting with 10% ethyl
acetate/hexanes to afford a light yellow oil (2.83 g). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 3.73-3.66 (m, 2H), 3.66 (s,
3H), 3.40-3.30 (m, 2H), 1.95-1.93 (dm, 1H), 1.92-1.90 (dm, 1H),
1.43 (ddd, J=13.74, 9.72, 3.89, 2H), 1.18 (s, 3H).
Cap-191, Step b
##STR00311##
[0630] To a solution of Cap-191, step a (3 g, 18.96 mmol) in
toluene (190 ml) at -78.degree. C. under nitrogen was added
diisobutylaluminum hydride (1.5M in toluene; 26.5 ml, 39.8 mmol)
dropwise. The reaction was continued to stir at -78.degree. C. for
1.5 h., and the bath was removed and was stirred for 18 h. The
reaction was quenched with MeOH (20 mL). HCl (1M, 150 mL) was added
and the mixture was extracted with EtOAc (4.times.40 mL). The
combined organic phases were washed with brine, dried (MgSO.sub.4),
filtered, and concentrated in vacuo. The residue was purified with
flash chromatography (silica gel; 40% ethyl acetate/hexanes) to
afford a colorless oil (1.36 g). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 3.77 (dt, J=11.73, 4.55, 2H), 3.69-3.60 (m, 2H), 3.42
(s, 2H), 1.71-1.40 (bs, 1H) 1.59 (ddd, J=13.74, 9.72, 4.39, 2H),
1.35-1.31 (m, 1H), 1.31-1.27 (m, 1H), 1.06 (s, 3H).
Cap-191, Step c
##STR00312##
[0632] To a solution of DMSO (5.9 ml, 83 mmol) in CH.sub.2Cl.sub.2
(85 ml) at -78.degree. C. under nitrogen was added oxalyl chloride
(3.8 ml, 43.4 mmol) and stirred for 40 min. A solution of Cap-191,
step b (4.25 g, 32.6 mmol) in CH.sub.2Cl.sub.2 (42.5 ml) was then
added. The reaction was continued to be stirred at -78.degree. C.
under nitrogen for 2 h. The reaction was quenched with cold 20%
K.sub.2HPO.sub.4 (aq) (10 mL) and water. The mixture was stirred at
.about.25.degree. C. for 15 min, diluted with diethyl ether (50 mL)
and the layers were separated. The aqueous layer was extracted with
diethyl ether (2.times.50 mL). The combined organic layers were
washed with brine, dried (MgSO.sub.4), filtered, and concentrated
in vacuo. The residue was taken up in CH.sub.2Cl.sub.2 (4 mL) and
purified with flash chromatography (silica gel, eluting with
CH.sub.2Cl.sub.2) to afford a colorless oil (2.1 g). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 9.49 (s. 1H), 3.80 (dt, J=11.98,
4.67, 2H), 3.53 (ddd, J=12.05, 9.41, 2.89, 2H), 1.98 (ddd, J=4.71,
3.20, 1.38, 1H), 1.94 (ddd, J=4.71, 3.20, 1.38, 1H), 1.53 (ddd,
J=13.87, 9.60, 4.14, 2H), 1.12 (s, 3H).
Cap-191, Step d
##STR00313##
[0634] To a solution of Cap 191c (2.5 g, 19.51 mmol) in CHCl.sub.3
(20 ml) under nitrogen at .about.25.degree. C. was added
(R)-2-amino-2-phenylethanol (2.94 g, 21.46 mmol) and stirred for 5
h. The reaction was cooled to 0.degree. C., trimethylsilyl cyanide
(3.8 ml, 30.4 mmol) was added dropwise. The cold bath was removed
and the reaction was allowed to stir at .about.25.degree. C. under
nitrogen for 15.5 h. The reaction was treated with 3N HCl (20 mL)
and water (20 mL), and the product was extracted with CHCl.sub.3
(3.times.50 mL). The combined organic layers were dried
(NaSO.sub.4), filtered, and concentrated in vacuo. The residue was
purified with flash chromatography (silica gel; 40% ethyl
acetate/hexanes) to afford two diastereomers: Cap-191, step d1
(diastereomer 1) as a colorless oil which solidified into a white
solid upon standing (3 g). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 7.42-7.26 (m, 5H), 5.21 (t, J=5.77, 1H), 3.87 (dd,
J=8.53, 4.52, 1H), 3.61-3.53 (m, 1H), 3.53-3.37 (m, 5H), 3.10 (d,
J=13.05, 1H), 2.65 (d, J=13.05, 1H), 1.64-1.55 (m, 1H), 1.55-1.46
(m, 1H), 1.46-1.39 (m, 1H), 1.31-1.23 (m, 1H), 1.11 (s, 3H). LC-MS:
Anal. Calcd. for [M+H].sup.+ C.sub.16H.sub.23N.sub.2O.sub.2:
275.18; found 275.20. Cap-191, step d2 (diastereomer 2) as a light
yellow oil (0.5 g). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
7.44-7.21 (m, 5H), 4.82 (t, J=5.40, 1H), 3.82-3.73 (m, 1H),
3.73-3.61 (m, 3H), 3.61-3.37 (m, 5H), 2.71 (dd, J=9.29, 4.77, 1H),
1.72-1.55 (m, 2H), 1.48-1.37 (m, 1H), 1.35-1.25 (m, 1H), 1.10 (s,
3H). LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.16H.sub.23N.sub.2O.sub.2: 275.18; found 275.20.
Cap-191, Step e
##STR00314##
[0636] To a solution of Cap-191, step d2 (diastereomer 2) (0.4472
g, 1.630 mmol) in CH.sub.2Cl.sub.2 (11 ml) and MeOH (5.50 ml) at
0.degree. C. under nitrogen was added lead tetraacetate (1.445 g,
3.26 mmol). The reaction was stirred for 1.5 h, the cold bath was
removed and stirring was continued for 20 h. The reaction was
treated with a phosphate buffer (pH=7; 6 mL) and stirred for 45
min. The reaction was filtered over CELITE.RTM., washed with
CH.sub.2Cl.sub.2 and the layers were separated. The aqueous layer
was extracted with CH.sub.2Cl.sub.2 (3.times. 25 mL), and the
combined organic layers was washed with brine, dried (MgSO.sub.4),
filtered and concentrated in vacuo. The residue was purified with
flash chromatography (silica gel; 15% ethyl acetate/hexanes) to
afford the imine intermediate as a colorless oil (181.2 mg).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 8.55 (d, J=1.00,
1H), 7.89-7.81 (m, 2H), 7.61-7.46 (m, 3H), 4.80 (d, J=1.00, 1H),
3.74 (tt, J=11.80, 4.02, 2H), 3.62-3.46 (m, 2H), 1.79-1.62 (m, 2H),
1.46-1.30 (m, 2H), 1.15 (s, 3H).
[0637] The imine intermediate was taken up in 6N HCl (10 mL) and
heated at 90.degree. C. for 10 days. The reaction was removed from
the heat, allowed to cool to room temperature and extracted with
ethyl acetate (3.times.25 mL). The aqueous layer was concentrated
in vacuo to afford an off-white solid. The solid was taken up in
MeOH and loaded on a pre-conditioned MCX (6 g) cartridge, washed
with MeOH followed by elution with 2N NH.sub.3/MeOH solution and
concentrated in vacuo to afford an off-white solid (79.8 mg).
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 14.33-13.51 (bs,
1H), 8.30 (bs, 3H), 3.82-3.75 (m, 1H), 3.70 (dt, J=11.80, 4.02,
2H), 3.58-3.43 (m, 2H), 1.76-1.60 (m, 2H), 1.47-1.36 (m, 1H),
1.36-1.27 (m, 1H), 1.08 (s, 3H). LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.8H.sub.16NO.sub.3: 174.11; found 174.19.
Cap-191 (Enantiomer-1)
[0638] To a solution of Cap-191, step e (0.0669 g, 0.386 mmol) and
sodium carbonate (0.020 g, 0.193 mmol) in sodium hydroxide (1M aq.;
0.4 ml, 0.40 mmol) at 0.degree. C. was added methyl chloroformate
(0.035 ml, 0.453 mmol) dropwise. The reaction was removed from the
cold bath and allowed to stir at .about.25.degree. C. for 3 h. The
reaction was washed with diethyl ether (3.times.20 mL). The aqueous
layer was acidified with 12 N HCl (pH .about.1-2), and extracted
with ethyl acetate (2.times.20 mL). The combined organic layers
were dried (MgSO.sub.4), filtered, and concentrated in vacuo to
afford Cap-191 as a colorless film (66.8 mg). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 13.10-12.37 (bs, 1H), 7.37 (d, J=9.04,
1H), 4.02 (d, J=9.29, 1H), 3.72-3.57 (m, 2H), 3.56 (s, 3H),
3.54-3.44 (m, 2H), 1.65 (ddd, J=13.61, 9.72, 4.27, 1H), 1.53 (ddd,
J=13.68, 9.66, 4.27, 1H), 1.41-1.31 (m, 1H), 1.31-1.22 (m, 1H),
1.00 (s, 3H). LC-MS: Anal. Calcd. for [M+Na].sup.+
C.sub.10H.sub.17NO.sub.5Na: 254.10; found 254.11.
Cap-192 (Enantiomer-2)
##STR00315##
[0640] Cap-192 (Enantiomer-2) was prepared from Cap-191, step d1
according to the procedure described for the preparation of its
enantiomer Cap-191.
INTERMEDIATES
##STR00316## ##STR00317##
##STR00318##
[0641] Literature ChemComm, 2172-2174 (2005).
Mixture of Intermediate 1
2,6-Bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
And Intermediate 2
2,7-Bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
[0642] To a degassed solution of naphthalene (19.34 g, 151 mmol)
and bis(pinacolato)diboron (84 g, 330 mmol) in cyclohexane (500 mL)
was added 4,4'-di-tert-buthyl-2,2'-dipyridyl (4.05 g, 15.1 mmol)
and di-.mu.-methoxybis(1,5-cyclooctadiene)diirridium (I) (5.0 g,
7.5 mmol). The flask was sealed and heated at 80.degree. C. for 16
h then allowed to cool to room temperature. The reaction color
turned dark red upon heating. The volatile component was removed in
vacuo and the resulting material was purified with flash
chromatography (sample was dry loaded on silica gel and eluted with
0-100% ethyl acetate/toluene) to afford a mixture of
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene and
2,7-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(54.57 g, 95% yield, 1.5:1 ratio) as white solid.
[0643] Intermediate 1: .sup.1H NMR (500 MHz, benzene-d.sub.6)
.delta. ppm 8.70 (s, 2H), 8.16 (d, J=8.2 Hz, 2H), 7.75 (d, J=8.2
Hz, 2H), 1.15 (s, 24H).
[0644] Intermediate 2: .sup.1H NMR (500 MHz, benzene-d.sub.6)
.delta. ppm 8.77 (s, 2H), 8.22 (d, J=8.2 Hz, 2H), 7.66 (d, J=8.2
Hz, 2H). 1.15 (s, 24H).
##STR00319##
Mixture of Intermediate 3
(2S,2'S)-tert-Butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyr-
rolidine-1-carboxylate
And Intermediate 4
(2S,2'S)-tert-Butyl
2,2'-(5,5'-(naphthalene-2,7-diyl)bis(1H-benzo[d]imidazole-5,2-diyl))dipyr-
rolidine-1-carboxylate
[0645] To a mixture of
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene and
2,7-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(1.5:1) (1.25 g, 3.29 mmol) and (S)-tert-butyl
2-(6-bromo-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate (3.01
g, 8.22 mmol) in 1,2-dimethoxyethane (50 mL) and water (12.5 mL)
was added NaHCO.sub.3 (1.38 g, 16.4 mmol). The reaction mixture was
degassed in vacuo and flushed with nitrogen. Pd(Ph.sub.3P).sub.4
(190 mg, 0.164 mmol) was added and the pressure flask was capped
and heated with an oil bath at 85.degree. C. for 16 hours. The
volatile component was removed in vacuo. The residue was
partitioned between ethyl acetate and water. The layers were
separated and the aqueous phase was extracted several times with
ethyl acetate. The combined organic phases were filtered through a
pad of diatomaceous earth (CELITE.RTM.) and the filtrate was dried
(Na.sub.2SO.sub.4), filtered and concentrated in vacuo. The
resulting material was purified with flash chromatography (sample
was dry loaded on silica gel and eluted with 10-100% ethyl
acetate/CH.sub.2Cl.sub.2) then flushed with (10%
methanol/CH.sub.2Cl.sub.2) to afford a mixture of
(2S,2'S)-tert-butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyr-
rolidine-1-carboxylate and (2S,2'S)-tert-butyl
2,2'-(5,5'-(naphthalene-2,7-diyl)bis(1H-benzo[d]imidazole-5,2-diyl))dipyr-
rolidine-1-carboxylate (622 mg, 27.1% yield) as orange solid which
was used without further purification. An aliquot was purified by a
reverse phase HPLC (water/acetonitrile/TFA) to provide an
analytical sample of each intermediate as a TFA salt.
[0646] Analytical data for Intermediate 3: LC-MS retention time
1.46 min; calcd. for C.sub.42H.sub.46N.sub.6O.sub.4: 698.36; found
m/z 699.26 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (TFA salt, 500 MHz,
MeOD) .delta. ppm 8.28 (br s, 2H), 8.13 (t, J=8.7 Hz, 4H), 8.05 (t,
J=8.4 Hz, 2H), 7.92 (dd, J=14.8, 8.7 Hz, 3H), 5.20-5.36 (m, 2H),
3.70-3.84 (m, 2H), 3.57-3.70 (m, 2H), 2.54-2.70 (m, 2H), 2.18-2.28
(m, 2H), 2.14 (quin, J=6.9 Hz, 4H), 1.50 (s, 9H), 1.23 (s, 9H).
[0647] Analytical data for Intermediate 4: LC-MS retention time
1.52 min; calcd. for C.sub.42H.sub.46N.sub.6O.sub.4: 698.36; found
m/z 699.24 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (TFA salt, 500 MHz,
MeOD) .delta. ppm 8.33 (br s, 2H), 7.98-8.17 (m, 6H), 7.83-7.98 (m,
4H), 5.18-5.35 (m, 2H), 3.71-3.83 (m, 2H), 3.57-3.70 (m, 2H), 2.63
(br s, 2H), 2.19-2.28 (m, 2H), 2.07-2.18 (m, 4H), 1.50 (s, 9H),
1.22 (s, 9H).
##STR00320##
Mixture of Intermediate 5
(S)-tert-Butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate
trifluoroacetate
And Intermediate 6
(S)-tert-Butyl
2-(6-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate
trifluoroacetate
[0648] After flash chromatography purification, the above reaction
also provided a mixture of (S)-tert-butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate trifluoroacetate
and (S)-tert-butyl
2-(6-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate (658 mg). During
the HPLC separation (TFA buffer) the boronic esters partially
hydrolyzed to afford the same mixture as boronic acid
intermediates. LC-MS retention time 1.34 min (boronic acids) and
2.05 (boronic esters); calcd. for (boronic acids)
C.sub.26H.sub.28BN.sub.3O.sub.4: 457.22 Found m/z 458.14
[M+H].sup.+. For (boronic esters) [M+H].sup.+
C.sub.32H.sub.38BN.sub.3O.sub.4: 539.3; found m/z 490.16 [M].sup.+.
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Waters Sunfire 5u C18 4.6.times.50 mm column using
a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min and an analysis time
of 4 min where Solvent A was 10% acetonitrile/90% water/0.1% TFA
and Solvent B was 90% acetonitrile/10% water/0.1% TFA.
##STR00321##
Intermediate 7
2,6-Bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
And Intermediate 8
2,7-Bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
[0649] A mixture of (2S,2'S)-tert-butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyr-
rolidine-1-carboxylate and (2S,2'S)-tert-butyl
2,2'-(5,5'-(naphthalene-2,7-diyl)bis(1H-benzo[d]imidazole-5,2-diyl))dipyr-
rolidine-1-carboxylate (594 mg, 0.162 mmol) and TFA (2 mL) in
CH.sub.2Cl.sub.2 (10 mL) was stirred at ambient conditions for 3
hours. The volatile component was removed in vacuo and the crude
material was purified by a reverse phase HPLC
(water/acetonitrile/TFA) to provide a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(434 mg) as yellow solid and a TFA salt of
2,7-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(204.8 mg) as yellow solid.
[0650] Analytical data for Intermediate 7: LC-MS retention time
1.05 min; calcd. for C.sub.32H.sub.30N.sub.6 498.25; found m/z
499.21 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.18 (s, 1H), 8.06 (d, J=8.6 Hz, 1H), 8.00 (s, 1H),
7.84-7.92 (m, 1H), 7.73-7.81 (m, 2H), 5.08 (t, J=7.6 Hz, 1H),
3.56-3.66 (m, 1H), 3.48-3.56 (m, 1H), 2.62-2.71 (m, 1H), 2.36-2.47
(m, 1H), 2.19-2.36 (m, 2H).
[0651] Analytical data for Intermediate 8: LC-MS retention time
1.11 min; calcd. for C.sub.32H.sub.30N.sub.6 498.25 Found m/z
499.20 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.23 (s, 2H), 8.01 (m, 4H), 7.82-7.89 (m, 2H),
7.69-7.82 (m, 4H), 5.08 (t, J=7.6 Hz, 2H), 3.57-3.66 (m, 2H),
3.48-3.57 (m, 2H), 2.60-2.73 (m, 2H), 2.37-2.48 (m, 2H), 2.19-2.36
(m, 4H).
##STR00322## ##STR00323##
##STR00324##
Intermediate 9
(S)-tert-Butyl 2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0652] Glyoxal (2.0 mL of 40% in water) was added dropwise over 11
minutes to a methanol solution of NH.sub.4OH (32 mL) and
(S)-Boc-prolinal (8.56 g, 43.0 mmol) and the reaction was stirred
at ambient temperature for 19 hours. The volatile component was
removed in vacuo and the residue was purified by a flash
chromatography (silica gel, ethyl acetate) followed by a
recrystallization (ethyl acetate, room temperature) to provide
(S)-tert-butyl 2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate as a
white fluffy solid (4.43 g, 18.6 mmol, 43% yield). .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. ppm 11.68/11.59 (br s, 1H), 6.94
(s, 1H), 6.76 (s, 1H), 4.76 (m, 1H), 3.48 (m, 1H), 3.35-3.29 (m,
1H), 2.23-1.73 (m, 4H), 1.39/1.15 (s, 9H). LC-MS. RT=0.87 min;
>95% homogeneity index; LC-MS: Anal. Calcd. for [M+H].sup.+
C.sub.12H.sub.20N.sub.3O.sub.2 238.16; found 238.22. The compound
shown to have a 98.9 ee % when analyzed under the chiral HPLC
conditions noted below.
Column. CHIRALPAK.RTM. AD, 10 um, 4.6.times.50 mm Solvent: 1.7%
ethanol/heptane (isocratic) Flow rate: 1 mL/min Wavelength: either
220 or 256 nm Relative retention time: 3.25 min (R), 5.78 minutes
(S)
##STR00325##
(S)-tert-Butyl
2-(4,5-diiodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0653] Iodine (16.17 g, 63.7 mmol) was added to a solution of
(S)-tert-butyl 2-(1H-imidazol-2-yl)pyrrolidine-1-carboxylate (6.87
g, 29.0 mmol) and sodium carbonate (9.21 g, 87 mmol) in dioxane (72
mL) and water (72 mL) at ambient temperature. The flask was covered
with aluminum foil and stirrer for 16 hours. The reaction mixture
was diluted with ethyl acetate and a saturated aqueous solution of
sodium thiosulfate. The mixture was stirred for 15 minutes and the
phases were separated. The aqueous phase was extracted several
times with ethyl acetate. The combined organic phases were dried
(Na.sub.2SO.sub.4), filtered and concentrated in vacuo to afford
(S)-tert-butyl
2-(4,5-diiodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (12.5 g
88%) as a tan solid. LC-MS retention time 1.40 min; calcd. for
C.sub.12H.sub.17I.sub.2N.sub.3O.sub.2 488.94; found m/z 489.96
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 4.72-4.84 (m, 1H),
3.58-3.70 (m, 1H), 3.43-3.54 (m, 1H), 2.36 (br s, 1H), 1.88-2.08
(m, 3H), 1.47 (br s, 3H), 1.27 (br s, 6H).
##STR00326##
(S)-tert-Butyl
2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0654] Sodium sulfite (10.31 g, 82 mmol) was added to a solution of
(S)-tert-butyl
2-(4,5-diiodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (4.0 g,
8.2 mmol) in ethanol (75 mL) and water (75 mL). The suspension was
heated with an oil bath at 100.degree. C. for 4 hours and at
90.degree. C. for 16 h. The reaction was diluted with ethyl acetate
and water. The layers were separated and the aqueous layer was
extracted several times with ethyl acetate. The combined organic
phases were dried (brine, Na.sub.2SO.sub.4), filtered and
concentrated in vacuo. The residue was purified by a flash
chromatography (sample was dry loaded on silica gel and eluted
with, 0 to 40% ethyl acetate/CH.sub.2Cl.sub.2) to afford
(S)-tert-butyl 2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(2.17 g, 73.1%) as a white solid. LC-MS retention time 0.930 min;
calcd. for C.sub.12H.sub.18IN.sub.3O.sub.2 363.04; found m/z 364.06
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.52-7.64 (m, 1H),
4.95-5.10 (m, 1H), 3.57-3.70 (m, 1H), 3.47-3.57 (m, 1H), 2.37-2.55
(m, 1H), 1.94-2.10 (m, 3H), 1.46 (s, 4H), 1.27 (s, 5H).
##STR00327##
##STR00328##
Mixture of Intermediate 12
(S)-tert-Butyl
2-(5-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imidaz-
ol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
And Intermediate 13
(S)-tert-Butyl
2-(5-(7-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imidaz-
ol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0655] To a mixture of (S)-tert-butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate trifluoroacetate
and (S)-tert-butyl
2-(6-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate trifluoroacetate
(560 mg, 0.980 mmol) and (S)-tert-butyl
2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (463 mg, 1.27
mmol) in 1,2-dimethoxyethane (11.2 mL) and water (2.8 mL) was added
NaHCO.sub.3 (412 mg, 4.90 mmol). The reaction mixture was degassed
in vacuo for 5 minutes and was flushed with nitrogen.
Pd(Ph.sub.3P).sub.4 (57 mg, 0.049 mmol) was added and the pressure
flask was capped and heated with an oil bath at 100.degree. C. for
16 hours. The volatile component was removed in vacuo. The residue
was partitioned between ethyl acetate and water and the aqueous
phase was extracted several times with ethyl acetate. The combined
organic phases were dried (Na.sub.2SO.sub.4), filtered and
concentrated in vacuo. The resulting material was purified with
flash chromatography (sample was dry loaded on silica gel and
eluted with 50-100% ethyl acetate/hexanes) to afford a partially
purified mixture of products which was further purified by a
reverse phase HPLC (water/acetonitrile/TFA) to provide a mixture of
a TFA salt of (S)-tert-butyl
2-(5-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imidaz-
ol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
and a TFA salt of (S)-tert-butyl
2-(5-(7-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imidaz-
ol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(74 mg). LC-MS retention time 1.32 min; calcd. for [M+H].sup.+
C.sub.38H.sub.44N.sub.6O.sub.4 648.34; found m/z 649.20
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. The mixture was used without further purification.
##STR00329##
Intermediate 14
2-((S)-Pyrrolidin-2-yl)-6-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)nap-
hthalen-2-yl)-1H-benzo[d]imidazole
And Intermediate 15
2-((S)-Pyrrolidin-2-yl)-6-(7-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-4-yl)nap-
hthalen-2-yl)-1H-benzo[d]imidazole
[0656] A mixture of (S)-tert-butyl
2-(5-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imidaz-
ol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
and (S)-tert-butyl
2-(5-(7-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imidaz-
ol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
as TFA salts (74 mg) and TFA (2 mL) in CH.sub.2Cl.sub.2 (10 mL) was
stirred at ambient conditions for 4 hours. The volatile component
was removed in vacuo. The crude material was purified by a reverse
phase HPLC (0 to 50% water/acetonitrile/TFA) to provide a TFA salt
of
2-((S)-pyrrolidin-2-yl)-6-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)na-
phthalen-2-yl)-1H-benzo[d]imidazole (32 mg, 31%) as yellow oil and
a TFA salt of
2-((S)-pyrrolidin-2-yl)-6-(7-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-
-4-yl)naphthalen-2-yl)-1H-benzo[d]imidazole (16 mg) as yellow
oil.
[0657] Analytical data for Intermediate 14: LC-MS retention time
0.081 min; calcd. for C.sub.38H.sub.28N.sub.6 448.24; found m/z
449.20 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.33 (s, 1H), 8.17 (s, 1H), 8.03 (d, J=3.7 Hz, 1H),
8.01 (s, 2H), 7.92-7.97 (m, 1H), 7.87 7.92 (m, 1H), 7.75-7.80 (m,
3H), 5.11 (t, J=7.8 Hz, 1H), 4.99 (t, J=7.9 Hz, 1H), 3.49-3.69 (m,
4H), 2.56-2.76 (m, 2H), 2.41-2.52 (m, 2H), 2.21-2.41 (m, 4H).
[0658] Analytical data for Intermediate 15: LC-MS retention time
0.87 min; calcd. for C.sub.38H.sub.28N.sub.6 448.24; found m/z
449.21 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.39 (s, 1H), 8.19 (s, 1H), 7.97-8.04 (m, 3H), 7.91 (d,
J=8.6 Hz, 1H), 7.87 (d, J=8.6 Hz, 1H), 7.85 (s, 1H), 7.78 (s, 2H),
5.12 (t, J=7.6 Hz, 1H), 5.04 (t, J=8.1 Hz, 1H), 3.50-3.68 (m, 4H),
2.60-2.74 (m, 2H), 2.41-2.55 (m, 2H), 2.21-2.41 (m, 4H).
##STR00330##
##STR00331##
(S)-tert-Butyl
2-(6-bromo-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate
[0659] EDCI.HCl (16.9 g, 88.0 mmol) was added to a mixture of
4-bromobenzene-1,2-diamine (15.0 g, 80.0 mmol), N-Boc-L-proline
(18.13 g, 84.0 mmol) and 1-hydroxybenzotriazole (12.28 g, 80.0
mmol) in CH.sub.2Cl.sub.2 (500 mL) and stirred at ambient
conditions for 16 h. The mixture was then diluted with water. The
resulting white precipitate was filtered away and the layers were
separated. The organic layer was washed with water, dried (brine;
Na.sub.2SO.sub.4), filtered and concentrated in vacuo to provide a
brown foam. Acetic acid (300 mL) was added to the foam and the
mixture was heated at 85.degree. C. (bath temperature) for 5 h. The
volatile component was removed in vacuo and the residue was
dissolved in EtOAc, washed with water and the organic phase was
dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo. The
resultant crude material was submitted to flash chromatography
(silica gel; 0-37% EtOAc/CH.sub.2Cl.sub.2). The partially pure
material was re-submitted to flash chromatography (silica gel;
20-35% EtOAc/CH.sub.2Cl.sub.2) to provide (S)-tert-butyl
2-(6-bromo-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate
(17.98 g, 61.2% yield) as yellow foam. LC-MS retention time 1.23
min; calcd. for C.sub.16H.sub.20BrN.sub.3O.sub.2: 365.07; found m/z
368.07 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 7.59-7.75 (m., 1H), 7.38-7.53 (m, 1H),
7.31-7.38 (m, 1H), 5.04-5.09 (m, 0.3H), 4.94-5.00 (m, 0.7H),
3.68-3.76 (m, 1H), 3.50-3.59 (m, 1H), 2.34-2.51 (m., 1H), 1.95-2.12
(m., 3H), 1.47 (br s, 3H), 1.15 (s, 6H).
##STR00332##
(S)-tert-Butyl
2-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)pyrrolidine-1-carboxylate
[0660] To a flask containing a mixture of (S)-tert-butyl
2-(6-bromo-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate (4.5
g, 12 mmol) and bis(pinacolato)diboron (6.55 g, 25.8 mmol) in
1,4-dioxane (50 mL) was added potassium acetate (3.01 g, 30.7
mmol). The reaction flask was degassed in vacuo for 5 minutes and
then purged with nitrogen. The catalyst Pd(Ph.sub.3P).sub.4 (710
mg, 0.614 mmol) was added and the flask was capped and heated with
an oil bath at 85.degree. C. (bath temperature) for 16 h. The
reaction mixture was concentrated in vacuo. The crude material was
partitioned between CH.sub.2Cl.sub.2 and a saturated NaHCO.sub.3
solution and the aqueous layer was extracted with CH.sub.2Cl.sub.2.
The combined organic phase was dried (brine, Na.sub.2SO.sub.4),
filtered and concentrated in vacuo. The resulting material was
purified with flash chromatography (sample was dry loaded on silica
gel and eluted with 10-50% ethyl acetate/CH.sub.2Cl.sub.2) to
provide (S)-tert-butyl
2-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)pyrrolidine-1-carboxylate (2.80 g, 55% yield) as white solid.
LC-MS retention time 1.493 min; calcd. for
C.sub.22H.sub.32BN.sub.3O.sub.4: 413.25; found m/z 414.23
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 7.81-8.08 (m, 1H),
7.40-7.68 (m., 2H), 4.95-5.13 (m, 1H), 3.68-3.79 (br s, 1H),
3.48-3.60 (br s, 1H), 2.35-2.52 (br s, 1H), 1.95-2.15 (m, 3H), 1.46
(s, 3H), 1.37 (s, 12H), 1.13 (s, 6H).
##STR00333##
tert-Butyl
2-(5-(6-(2-(1-(tert-butoxycarbonyl)-2-pyrrolidinyl)-1H-benzimidazol-6-yl)-
-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxylate
[0661] To a mixture of (S)-tert-butyl
2-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)pyrrolidine-1-carboxylate (1.29 g, 33.1 mmol) and
2,6-dibromonaphthalene (446 mg, 1.56 mmol) in 1,2-dimethoxyethane
(20 mL) and water (6 mL) was added NaHCO.sub.3 (787 mg, 9.36 mmol).
The reaction mixture was degassed in vacuo for 10 minutes and was
flushed with nitrogen. The catalyst Pd(Ph.sub.3P).sub.4 (90 mg,
0.078 mmol) was added and the flask was capped and heated with an
oil bath at 100.degree. C. for 16 hours. The volatile component was
removed in vacuo. The residue was partitioned between
CH.sub.2Cl.sub.2 and water and the layers were separated. The
aqueous phase was extracted several times with CH.sub.2Cl.sub.2 and
the combined organic phases were dried (brine, Na.sub.2SO.sub.4),
filtered and concentrated in vacuo. The resulting material was
purified with flash chromatography (sample was dry loaded on silica
gel and eluted with 0-70% ethyl acetate/CH.sub.2Cl.sub.2) to afford
tert-butyl
2-(5-(6-(2-(1-(tert-butoxycarbonyl)-2-pyrrolidinyl)-1H-benzimidazol-6-yl)-
-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxylate (605
mg) as yellow solid. LC-MS retention time 1.46 min; calcd. for
C.sub.42H.sub.46N.sub.6O.sub.4: 698.36; found m/z 699.26
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.28 (br s, 2H), 8.13
(t, J=8.7 Hz, 4H), 8.05 (t, J=8.4 Hz, 2H), 7.92 (dd, J=14.8, 8.7
Hz, 4H), 5.20-5.36 (m, 2H), 3.70-3.84 (m, 2H), 3.57-3.70 (m, 2H),
2.54-2.70 (m, 2H), 2.18-2.28 (m, 2H), 2.14 (quin, J=6.9 Hz, 4H),
1.50 (s, 9H), 1.23 (s, 9H).
##STR00334##
tert-Butyl
(2S)-2-(5-(6-bromo-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxy-
late
[0662] The above reaction also provided tert-butyl
(2S)-2-(5-(6-bromo-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxy-
late (519 mg) as yellow solid. LC-MS retention time 1.88 min;
calcd. for [M+H].sup.+ C.sub.26H.sub.26BrN.sub.3O.sub.2: 491.12;
found m/z 494.04 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.21 (d, J=8.6 Hz, 1H), 8.05-8.14 (m, 2H), 7.93-8.05
(m, 2H), 7.82-7.93 (m, 3H), 7.57-7.67 (m, 1H), 5.28 (br s, 1H),
3.98 (s, 1H), 3.70-3.80 (m, 1H), 3.57-3.70 (m, 1H), 2.54-2.71 (m,
1H), 2.17-2.29 (m, 1H), 2.09-2.17 (m, 2H), 1.49 (s, 4H), 1.22 (s,
5H).
##STR00335##
5,5'-(2,6-Naphthalenediyl)bis(2-(2-pyrrolidinyl)-1H-benzimidazole)
[0663] A mixture of tert-butyl
2-(5-(6-(2-(1-(tert-butoxycarbonyl)-2-pyrrolidinyl)-1H-benzimidazol-6-yl)-
-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxylate (591
mg. 0.844 mmol) and TFA (2 mL) in CH.sub.2Cl.sub.2 (10 mL) was
stirred at ambient conditions for 2 hours. The volatile component
was removed in vacuo and the resulting material was loaded onto a
MCX column, flushed with methanol, released with 2.0 M
NH.sub.3/methanol elution) and concentrated to provide
5,5'-(2,6-naphthalenediyl)bis(2-(2-pyrrolidinyl)-1H-benzimidazole)
(419 mg) as tan solid. LC-MS retention time 1.02 min; calcd. for
C.sub.33H.sub.31N.sub.5: 498.25; found m/z 499.25 [M+H].sup.+. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Waters Sunfire 5u C18 4.6.times.50 mm column using
a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min and an analysis time
of 4 min where Solvent A was 10% acetonitrile/90% water/0.1% TFA
and Solvent B was 90% acetonitrile/10% water/0.1% TFA. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.18 (s, 1H), 8.06 (d, J=8.6 Hz, 1H),
8.00 (s, 1H), 7.84-7.92 (m, 1H), 7.73-7.81 (m, 2H), 5.08 (t, J=7.6
Hz, 1H), 3.56-3.66 (m, 1H), 3.48-3.56 (m, 1H), 2.62-2.71 (m, 1H),
2.36-2.47 (m, 1H), 2.19-2.36 (m, 2H).
##STR00336##
##STR00337##
2,6-Bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
[0664] A pressure flask containing a mixture of
2,6-dibromonaphthalene (8.0 g, 28 mmol), bis(pinacolato)diboron
(21.3 g, 84.0 mmol), potassium acetate (3.01 g, 30.7 mmol) and
1,4-dioxane (50 mL) was degassed in vacuo for 5 minutes and purged
with nitrogen. The catalyst Pd(Ph.sub.3P).sub.4 (710 mg, 0.614
mmol) was added. The flask was capped and heated with an oil bath
at 85.degree. C. (bath temperature) for 16 hours. The reaction
mixture was concentrated in vacuo and the crude material was
partitioned between CH.sub.2Cl.sub.2 and a saturated NaHCO.sub.3
solution. The aqueous layer was extracted with CH.sub.2Cl.sub.2 and
the combined organic phases was dried (Na.sub.2SO.sub.4), filtered
and concentrated in vacuo. The resulting material was purified with
flash chromatography (sample was dry loaded on silica gel and
eluted with 50-100% CH.sub.2Cl.sub.2/Hexanes) to provide
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
contaminated with bis(pinacolato)diboron. The solid was dissolved
into CH.sub.2Cl.sub.2 and methanol was added until precipitation
was observed. The precipitate was collected by filtration to afford
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(5.1 g, 48% yield) as white solid. .sup.1H NMR (500 MHz,
benzene-d.sub.6) .delta. ppm 8.70 (s, 2H), 8.16 (d, J=8.2 Hz, 2H),
7.75 (d, J=8.2 Hz, 2H), 1.15 (s, 24H).
##STR00338##
(2S,2'S)-tert-Butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyr-
rolidine-1-carboxylate
[0665] To a mixture of
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(2.0 g, 5.3 mmol) and (S)-tert-butyl
2-(6-bromo-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate (2.89
g, 7.89 mmol) in 1,2-dimethoxyethane (42.1 mL) and water (10.5 mL)
was added NaHCO.sub.3 (2.21 g, 26.3 mmol). The reaction mixture was
degassed in vacuo and was flushed with nitrogen. The catalyst
Pd(Ph.sub.3P).sub.4 (90 mg, 0.078 mmol) was added. The flask was
capped and heated with an oil bath at 100.degree. C. for 16 hours.
The volatile component was removed in vacuo and the residue was
partitioned between ethyl acetate and water. The layers were
separated and the aqueous phase was extracted several times with
ethyl acetate. The combined organic phases were filtered through a
pad of diatomaceous earth (CELITE.RTM.) and the filtrate was dried
(Na.sub.2SO.sub.4), filtered and concentrated in vacuo. The
resulting material was purified with flash chromatography (sample
was dry loaded on silica gel and eluted with 0-100% ethyl
acetate/hexanes) to afford (2S,2'S)-tert-butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyr-
rolidine-1-carboxylate (761 mg, 21% yield) as orange solid. LC-MS
retention time 1.46 min; calcd. for C.sub.42H.sub.46N.sub.6O.sub.4:
698.36; found m/z 699.26 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA .sup.1H NMR (500 MHz,
MeOD) .delta. ppm 8.28 (br s, 2H), 8.13 (t, J=8.7 Hz, 4H), 8.05 (t,
J=8.4 Hz, 2H), 7.92 (dd, J=14.8, 8.7 Hz, 4H), 5.20-5.36 (m, 2H),
3.70-3.84 (m, 2H), 3.57-3.70 (m, 2H), 2.54-2.70 (m, 2H), 2.18-2.28
(m, 2H), 2.09-2.19 (m, 4H), 1.50 (s, 9H), 1.23 (s, 9H).
##STR00339##
(S)-tert-Butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate
[0666] The above reaction also provided (S)-tert-butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate (1.58 g, 56% yield)
as tan solid. LC-MS retention time 1.46 min; calcd. for
C.sub.32H.sub.38BN.sub.3O.sub.4: 539.3; found m/z 540.24
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA.
##STR00340##
##STR00341##
(S)-5-((tert-Butyldiphenylsilyloxy)methyl)pyrrolidin-2-one
[0667] To a solution of (S)-5-(hydroxymethyl)pyrrolidin-2-one (10
g, 87 mmol) in CH.sub.2Cl.sub.2 (50 mL) was added
tert-butylchlorodiphenylsilane (25.6 g, 93 mmol), Et.sub.3N (12.1
mL, 87 mmol) and DMAP (1.06 g, 8.7 mmol). The mixture was stirred
at room temperature until the starting pyrrolidinone was completely
consumed and then diluted with CH.sub.2Cl.sub.2 (50 mL) and washed
with water (50 mL). The organic layer was dried (Na.sub.2SO.sub.4),
filtered, evaporated in vacuo and the crude material was submitted
to flash chromatography (silica gel; 30 to 100% of EtOAc/hexanes)
to afford
(S)-5-((tert-butyldiphenylsilyloxy)methyl)pyrrolidin-2-one (22.7 g,
74% yield) as a colorless oil. LC-MS (M+H).sup.+=354.58. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.69 (br s, 1H), 7.64-7.61
(m, 4H), 7.50-7.42 (m, 6H), 3.67-3.62 (m, 1H), 3.58-3.51 (m, 2H),
2.24-2.04 (m, 3H), 1.87-1.81 (m, 1H), 1.00 (s, 9H).
##STR00342##
(S)-tert-Butyl
2-((tert-butyldiphenylsilyloxy)methyl)-5-oxopyrrolidine-1-carboxylate
[0668] Di-tert-butyl dicarbonate (38.5 g, 177 mmol) was added in
portions as a solid over 10 min to a solution of
(S)-5-((tert-butyldiphenylsilyloxy)methyl)pyrrolidin-2-one (31.2 g,
88.3 mmol), Et.sub.3N (8.93 g, 88 mmol) and DMAP (1.08 g, 8.83
mmol) in CH.sub.2Cl.sub.2 (200 mL) and the reaction mixture was
stirred for 18 h at 24.degree. C. Most of the volatile material was
removed in vacuo and the crude material was purified by silica gel
chromatography (20% EtOAc/hexanes to 50% EtOAc/hexanes) to afford
(S)-tert-butyl
2-((tert-butyldiphenylsilyloxy)methyl)-5-oxopyrrolidine-1-carboxylate
(32.65 g, 82% yield) as a white solid. LC-MS
(M-Boc+H).sup.+=354.58. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 7.61-7.59 (m, 2H), 7.56-7.54 (m, 2H), 7.50-7.38 (m, 6H), 4.18
(m, 1H), 3.90 (dd, J=10.4, 3.6, 1H), 3.68 (dd, J=10.4, 2.1, 1H),
2.68-2.58 (m, 1H), 2.40-2.33 (m, 1H), 2.22-2.12 (m, 1H), 2.01-1.96
(m, 1H), 1.35 (s, 9H), 0.97 (s, 9H).
##STR00343##
Intermediate 21
(S)-tert-Butyl
2-((tert-butyldiphenylsilyloxy)methyl)-2,3-dihydro-1H-pyrrole-1-carboxyla-
te
[0669] A three-necked flask equipped with a thermometer and a
nitrogen inlet was charged with (S)-tert-butyl
2-((tert-butyldiphenylsilyloxy)methyl)-5-oxopyrrolidine-1-carboxylate
(10.05 g, 22.16 mmol) and toluene (36 mL) and lowered into
-55.degree. C. cooling bath. When the internal temperature of the
mixture reached -50.degree. C., lithium triethylborohydride (23 mL
of 1.0 M/THF, 23 mmol) was added dropwise over 30 min and the
mixture stirred for 35 min while maintaining the internal
temperature between -50.degree. C. and -45.degree. C. Hunig's base
(16.5 mL, 94 mmol) was added dropwise over 10 min. Then, DMAP (34
mg, 0.278 mmol) was added in one batch, followed by the addition of
trifluoroacetic anhydride (3.6 mL, 25.5 mmol) over 15 min, while
maintaining the internal temperature between -50.degree. C. and
-45.degree. C. The bath was removed 10 min later and the reaction
mixture was stirred for 14 h while allowing it to rise to ambient
temperature. The reaction mixture was diluted with toluene (15 mL),
cooled with an ice-water bath and treated slowly with water (55 mL)
over 5 min. The phases were separated and the organic layer washed
with water (50 mL, 2.times.) and concentrated in vacuo. The crude
material was purified by flash chromatography (silica gel; 5%
EtOAc/hexanes) to afford (S)-tert-butyl
2-((tert-butyldiphenylsilyloxy)methyl)-2,3-dihydro-1H-pyrrole-1-carboxyla-
te (7.947 g, 82% yield) as a colorless viscous oil. LC-MS:
[M+Na].sup.+=460.19. Rt=2.41 min under the following HPLC
conditions: Solvent gradient from 100% A:0% B to 0% A:100% B
(A=0.1% TFA in 1:9 MeOH/water; B=0.1% TFA in 9:1 MeOH/water) over 2
min and hold for 1 min; detection at 220 nm; PHENOMENEX.RTM. Luna
3.0.times.50 mm S10 column. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 7.62-7.58 (m, 4H), 7.49-7.40 (m, 6H), 6.47 (br s, 1H),
5.07/5.01 (overlapping br d, 1H), 4.18 (br s, 1H), 3.89 (br s,
0.5H), 3.69 (br s, 1.5H), 2.90-2.58 (br m, 2H), 1.40/1.26
(overlapping br s, 9H), 0.98 (s, 9H).
##STR00344##
Intermediate 22
(3S)-tert-Butyl
3-((tert-butyldiphenylsilyloxy)methyl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate
[0670] Diethylzinc (19 mL of .about.1.1 M in toluene, 21 mmol) was
added dropwise over 15 min to a cooled (-30.degree. C.) toluene (27
mL) solution of (S)-tert-butyl
2-((tert-butyldiphenylsilyloxy)methyl)-2,3-dihydro-1H-pyrrole-1-carboxyla-
te (3.94 g, 9.0 mmol). Chloroiodomethane (stabilized over copper;
3.0 mL, 41 mmol) was added dropwise over 10 min and stirred while
maintaining the bath temperature at -25.degree. C. for 1 h and
between -25.degree. C. and -21.degree. C. for 18.5 h. The reaction
mixture was opened to the air and quenched by the slow addition of
50% saturated NaHCO.sub.3 solution (40 mL) and then removed from
the cooling bath and stirred at ambient temperature for 20 min. The
reaction mixture was filtered through a filter paper and the white
cake was washed with 50 mL of toluene. The organic phase of the
filtrate was separated and washed with water (40 mL, 2.times.),
dried (MgSO.sub.4) and concentrated in vacuo. The crude material
was purified using by silica gel chromatography (350 g silica gel;
sample was loaded with 7% EtOAc/hexanes; eluted with 7-20%
EtOAc/hexanes) to afford (3S)-tert-butyl
3-((tert-butyldiphenylsilyloxy)methyl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate (3.69 g, 90.7%) as a mixture of cis/trans-isomers. [Note: the
exact cis/trans-isomer ratio was not determined at this stage].
LC-MS: [M+Na].sup.+=474.14. Rt=2.39 min under the following HPLC
conditions: Solvent gradient from 100% A:0% B to 0% A:100% B
(A=0.1% TFA in 1:9 MeOH/water; B=0.1% TFA in 9:1 MeOH/water) over 2
min and hold for 1 min; detection at 220 nm; PHENOMENEX.RTM. Luna
3.0.times.50 mm S10 column. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 7.62-7.60 (m, 4H), 7.49-7.40 (m, 6H), 3.77/3.67
(overlapping br s, 3H), 3.11-3.07 (m, 1H), 2.23 (app br s, 1H),
2.05-2.00 (m, 1H), 1.56-1.50 (m, 1H), 1.33 (very broad s, 9H), 1.00
(s, 9H), 0.80 (m, 1H), 0.30 (m, 1H).
##STR00345##
Intermediate 23
(3S)-tert-Butyl
3-(hydroxymethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0671] TBAF (7.27 mL of 1.0 M in THF, 7.27 mmol) was added dropwise
over 5 min to a THF (30 mL) solution of (3S)-tert-butyl
3-((tert-butyldiphenylsilyloxy)methyl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate (mixture of cis/trans-isomers) (3.13 g, 6.93 mmol) and the
mixture was stirred at ambient temperature for 4.75 h. After the
addition of saturated NH.sub.4Cl solution (5 mL), most of the
volatile material was removed in vacuo and the residue partitioned
between CH.sub.2Cl.sub.2 (70 mL) and 50% saturated NH.sub.4Cl
solution (30 mL). The aqueous phase was extracted with
CH.sub.2Cl.sub.2 (30 mL) and the combined organic phase was dried
(MgSO.sub.4), filtered, concentrated in vacuo and then exposed to
high vacuum overnight. The crude material was purified using a
flash chromatography (silica gel; 40-50% EtOAc/hexanes) to afford
(3S)-tert-butyl
3-(hydroxymethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (1.39 g,
94% yield) as a mixture of cis/trans-isomers and a colorless oil.
[Note: the exact cis/trans-isomer ratio was not determined at this
stage.] LC-MS (M+Na).sup.+=236.20. .sup.1H NMR (400 MHz,
DMSO-d.sub.6, .delta.=2.5 ppm) .delta. ppm 4.70 (t, J=5.7, 1H),
3.62-3.56 (m, 1H), 3.49-3.44 (m, 1H), 3.33-3.27 (m, 1H), 3.08-3.04
(m, 1H), 2.07 (br m, 1H), 1.93-1.87 (m, 1H), 1.51-1.44 (m, 1H),
1.40 (s, 9H), 0.76-0.71 (m, 1H), 0.26 (m, 1H).
##STR00346##
Intermediate 24
(1R,3S,5R)-2-(tert-Butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid
[0672] A semi-solution of NaIO.sub.4 (6.46 g, 30.2 mmol) in water
(31 mL) was added to a solution of (3S)-tert-butyl
3-(hydroxymethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (mixture
of cis/trans-isomers) (2.15 g, 10.08 mmol) in CH.sub.3CN (20 mL)
and CCl.sub.4 (20 mL). RuCl.sub.3 (0.044 g, 0.212 mmol) was added
immediately and the heterogeneous reaction mixture was stirred
vigorously for 75 min. The reaction mixture was diluted with water
(60 mL) and extracted with CH.sub.2Cl.sub.2 (50 mL, 3.times.). The
combined organic phases was treated with 1 mL MeOH, allowed to
stand for about 5 min and then filtered through a pad of
diatomaceous earth (CELITE.RTM.). The pad was washed with
CH.sub.2Cl.sub.2 (50 mL) and the filtrate was concentrated in vacuo
to afford a light charcoal-colored solid. The crude material was
dissolved in EtOAc (.about.10 mL) with heating and allowed to stand
at ambient temperature with seeding. About 15 min into the cooling
phase, a rapid crystal formation was observed. About 1 h later,
hexanes (.about.6 mL) was added and the mixture refrigerated
overnight (it did not appear that additional material precipitated
out). The mixture was filtered and washed with ice/water-cooled
hexanes/EtOAc (2:1 ratio; 20 mL) and dried under high vacuum to
afford the first crop of
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (off-white crystals, 1.222 g). The mother liquor was
concentrated in vacuo and the residue dissolved in .about.3 mL of
EtOAc with heating, allowed to stand at ambient temperature for 1 h
and then 3 mL hexanes was added and stored in a refrigerator for
.about.15 h. A second crop of
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid was retrieved similarly (grey crystals, 0.133 g), for a
combined yield of 59%. LC-MS [M+Na].sup.+=250.22. Rt=1.48 min under
the following HPLC conditions: Solvent gradient from 100% A:0% B to
0% A:100% B (A=0.1% TFA in 1:9 methanol/water; B=0.1% TFA in 9:1
methanol/water) over 3 min; detection at 220 nm; PHENOMENEX.RTM.
Luna 3.0.times.50 mm S10 column. MP (dec.) for the first
crop=147.5-149.5.degree. C. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 12.46 (s, 1H), 3.88 (app br s, 1H), 3.27 (app br s, 1H;
overlapped with water signal), 2.28 (br m, 1H), 2.07 (app br s,
1H), 1.56 (app s, 1H), 1.40/1.34 (two overlapped s, 9H), 0.71 (m,
1H), 0.45 (m, 1H). .sup.13C-NMR (100.6 MHz, DMSO-d.sub.6) 172.96,
172.60, 154.45, 153.68, 78.74, 59.88, 59.58, 36.91, 31.97, 31.17,
27.77, 27.52, 14.86, 14.53, 13.69. Anal. Calcd. for
C.sub.11H.sub.17NO.sub.4: C, 58.13; H, 7.54; N, 6.16. Found (for
first crop): C, 58.24; H, 7.84; N, 6.07. Optical rotation (10 mg/mL
in CHCl.sub.3): [.alpha.]D=-216 and -212 for the first and second
crop, respectively.
An Alternative Synthesis for Intermediate 24:
##STR00347##
[0674] (1R,3R,5R)-2-Azabicyclo[3.1.0]hexane-3-carboxamide
(.CH.sub.3SO.sub.3H) was prepared according to the procedure
described for the synthesis of its stereoisomer in patent WO
2004/052850.
##STR00348##
Intermediate 24
(1R,3S,5R)-2-(tert-Butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid
[0675] A 1 L round bottom flask equipped with a nitrogen inlet,
overhead agitator, thermocouple and heating mantle was charged with
50 g (225 mmol) (1R,3R,5R)-2-Azabicyclo[3.1.0]hexane-3-carboxamide
(.CH.sub.3SO.sub.3H) and 250 mL isopropanol. The resulting slurry
was then charged with 252 mL of 23 wt % NaOEt in EtOH (2.68 M, 675
mmol, 3.0 equiv) and stirred at 50.degree. C. for ca. 1 h. The
mixture was charged with 12.2 mL (675 mmol, 3 equiv) of water and
heated to 60.degree. C. The resulting slurry was allowed to stir at
60.degree. C. for ca. 18 h. The slurry was cooled to rt and charged
with 250 mL water and 98.2 g (450 mmol, 2.0 equiv)
di-t-butyldicarbonate. Ethanol and isopropanol were removed via
vacuum distillation and the aqueous mixture cooled to 0.degree. C.
The mixture was neutralized with 76 ml (456 mmol) 6M aqueous HCl
while maintaining an internal temperature<5.degree. C. The
product was extracted with 500 mL MTBE and the rich organic layer
was washed with 100 mL water. The clear solution was concentrated
down to 150 mL via vacuum distillation and the resulting slurry was
charged with 600 mL heptane while maintaining an internal
temperature>45.degree. C. The slurry was cooled to rt over ca.
30 min and allowed to stir at rt for ca. 2 h. The product was
filtered, washed with 250 mL 4:1 heptane:MTBE and dried under
vacuum at 70.degree. C. to give 40.5 g (178 mmol, 79% yield, 99.8
AP at 205 nm) of
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid: .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.48 (s, 1H),
4.02-3.80 (m, 1H), 3.45-3.15 (m, 1H), 2.40-2.19 (m, 1H), 2.19-2.0
(m, 1H), 1.70-1.50 (m, 1H), 1.50-1.20 (m, 9H), 0.83-0.60 (m, 1H),
0.33-0.55 (m, 1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.
173.7, 173.2, 155.0, 154.3, 79.4, 60.5, 60.2, 37.6, 32.6, 31.8,
28.4, 28.2, 15.6, 15.2, 14.4; HRMS calcd for
C.sub.11H.sub.18NO.sub.4 (M+H; ESL): 228.1236. Found: 228.1234.
##STR00349##
##STR00350##
Intermediate 25
(1R,3S,5R)-tert-Butyl
3-(6-bromo-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxyl-
ate
[0676] EDCI.HCl (2.65 g, 13.8 mmol) was added to a mixture of
4-bromobenzene-1,2-diamine 2.35 g, 12.6 mmol),
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (3.00 g, 13.2 mmol) and 1-hydroxybenzotriazole (1.93 g, 12.6
mmol) in CH.sub.2Cl.sub.2 (80 mL) and stirred at ambient conditions
for 16 h. The mixture was then diluted with CH.sub.2Cl.sub.2,
washed with water, dried (brine; MgSO.sub.4), filtered and
concentrated in vacuo to provide a brown foam. Acetic acid (80 mL)
was added to the foam and the mixture was heated at 75.degree. C.
(bath temperature) for 5 h. The volatile component was removed in
vacuo and the residue was dissolved in EtOAc, washed with saturated
NaHCO.sub.3 solution and the organic phase was dried (brine;
MgSO.sub.4), filtered and concentrated in vacuo. The resultant
crude material was submitted to flash chromatography (silica gel;
50-100% EtOAc/hexanes) to provide (1R,3S,5R)-tert-butyl
3-(6-bromo-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxyl-
ate (3.01 g, 7.96 mmol, 63.3% yield) as a light orange foam, which
was used without further purification. The reaction also yielded
847 mg of same product with lower purity. An aliquot of the
collected material was purified further by preparative HPLC
(C-18/30-100% CH.sub.3CN-water+0.1% NH.sub.4OAc) to achieve an
analytical sample. LC-MS retention time 1.248 min; calcd. for
C.sub.17H.sub.21BrN.sub.3O.sub.2: 378.08; found m/z 380.05
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.67
(br s, 1H), 7.43 (br s, 1H), 7.34 (d, J=8.6 Hz, 1H), 4.75 (br s,
1H), 3.62 (br s, 1H), 2.50-2.57-2.31 (m, 1H), 2.31 (dt, J=13.2, 6.7
Hz, 1H), 1.66-1.85 (m, 1H), 1.45 (br s, 3H), 1.11 (br s, 6H), 0.87
(dt, J=8.6, 5.8 Hz, 1H), 0.66 (br s, 1H).
##STR00351##
Intermediate 26
(1R,3S,5R)-tert-Butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0677] To a pressure flask containing a mixture of
(1R,3S,5R)-tert-butyl
3-(6-bromo-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxyl-
ate (2.0 g, 5.3 mmol) and bis(pinacolato)diboron (2.69 g, 10.6
mmol) in 1,4-dioxane (50 mL) was added potassium acetate (0.78 g,
7.9 mmol). The reaction flask was degassed in vacuo for 10 min and
purged with nitrogen. Pd(Ph.sub.3P).sub.4 (305 mg, 0.264 mmol) was
added to the reaction mixture and the flask was capped and heated
with an oil bath at 85.degree. C. (bath temperature) for 16 hours.
The reaction mixture was concentrated in vacuo and the crude
material was partitioned between CH.sub.2Cl.sub.2 (150 mL) and an
aqueous medium (50 mL water+10 mL saturated NaHCO.sub.3 solution).
The aqueous layer was extracted with CH.sub.2Cl.sub.2 and the
combined organic phase was dried (Na.sub.2SO.sub.4), filtered and
concentrated in vacuo. The resulting material was purified with
flash chromatography (sample was dry loaded on silica gel and
eluted with 20-100% ethyl acetate/hexanes) to provide
(1R,3S,5R)-tert-butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (1.74 g, 77% yield) as
white foam. An aliquot of the collected material was further
purified by preparative HPLC (C-18/30-100% CH.sub.3CN-water+0.1%
NH.sub.4OAc). LC-MS retention time 1.78 min; calcd. for
C.sub.23H.sub.32BN.sub.32 425.25; found m/z 426.21 [M+H].sup.+. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 4 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 5% acetonitrile/95%
water/10 mmol ammonium acetate and Solvent B was 95%
acetonitrile/5% water/10 mmol ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.94 (s, 1H), 7.61 (d,
J=7.9 Hz, 1H), 7.41-7.56 (m, 1H), 4.83 (br s, 1H), 3.64 (d, J=1.2
Hz, 1H), 2.51-2.70 (m, 1H), 2.33 (dt, J=13.4, 6.6 Hz, 1H),
1.67-1.84 (m, J=8.5, 6.10, 5.8, 5.8 Hz, 1H), 1.39-1.59 (br s, 3H),
1.37 (s, 12H), 1.10 (br s, 6H), 0.88 (dt, J=8.2, 5.8 Hz, 1H), 0.66
(br s, 1H).
##STR00352##
Intermediate 27
(1R,1'R,3S,3'S,5R,5'R)-tert-Butyl
3,3'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))bis(2-
-azabicyclo[3.1.0]hexane-2-carboxylate)
[0678] To a mixture of (1R,3S,5R)-tert-butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (1.50 g, 3.53 mmol) and
2,6-dibromonaphthalene (403 mg, 1.41 mmol) in 1,2-dimethoxyethane
(21.7 mL) and water (6.50 mL) was added NaHCO.sub.3 (356 mg, 4.23
mmol). The reaction mixture was degassed in vacuo for 10 minutes
and flushed with nitrogen. Pd(Ph.sub.3P).sub.4 (82 mg, 0.071 mmol)
was added and the reaction mixture was heated with an oil bath at
100.degree. C. for 16 hours and then the volatile component was
removed in vacuo. The residue was partitioned between
CH.sub.2Cl.sub.2 and water (30 mL) and the aqueous phase was
extracted several times with CH.sub.2Cl.sub.2. The combined organic
phases were washed with brine, dried (Na.sub.2SO.sub.4), filtered
and concentrated in vacuo. The resulting material was purified with
flash chromatography (sample was dry loaded on silica gel and
eluted with 0-70% ethyl acetate/CH.sub.2Cl.sub.2) to afford
(1R,1'R,3S,3'S,5R,5'R)-tert-butyl
3,3'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))bis(2-
-azabicyclo[3.1.0]hexane-2-carboxylate) (377 mg) as yellow solid.
LC-MS retention time 1.49 min; calcd. for
C.sub.44H.sub.46N.sub.6O.sub.4: 722.36; found m/z 723.26
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.26
(s, 2H), 8.12 (d, J=8.5 Hz, 2H), 8.09 (s, 2H), 8.00 (m, J=8.6 Hz,
2H), 7.93 (m, J=8.6 Hz, 2H), 7.86 (d, J=8.6 Hz, 2H), 5.01 (br s,
2H), 3.68 (br s, 2H), 2.76 (dd, J=13.3, 9.3 Hz, 2H), 2.42-2.54 (m,
2H), 1.87 (br s, 2H), 1.49 (br s, 9H), 1.19 (br s, 9H), 0.94 (dt,
J=8.3, 6.1 Hz, 2H), 0.79 (br s, 2H).
##STR00353##
Intermediate 28
(1R,3S,5R)-tert-Butyl
3-(6-(6-bromonaphthalen-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate
[0679] The above reaction also provided (1R,3S,5R)-tert-butyl
3-(6-(6-bromonaphthalen-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate (418 mg) as yellow solid. LC-MS retention
time 1.90 min; calcd. for C.sub.27H.sub.26BrN.sub.3O.sub.2: 503.12;
found m/z 506.05 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.20 (s, 1H), 8.12 (s, 1H), 8.05 (s,
1H), 7.93-8.00 (m, 2H), 7.86-7.93 (m, 2H), 7.84 (d, J=8.6 Hz, 1H),
7.63 (dd, J=8.6, 1.8 Hz, 1H), 5.00 (br s, 1H), 3.68 (br s, 1H),
2.75 (dd, J=13.4, 9.2 Hz, 1H), 2.41-2.54 (m, 1H), 1.87 (br s, 1H),
1.49 (br s, 5H), 1.17 (br s, 4H), 0.87-0.98 (m, 1H), 0.79 (br s,
1H).
##STR00354##
Intermediate 29
2,6-Bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-
-yl)naphthalene
[0680] A mixture of (1R,1'R,3S,3'S,5R,5'R)-tert-butyl
3,3'-(6,6'-(naphthalene-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))bis(2-
-azabicyclo[3.1.0]hexane-2-carboxylate) (360 mg. 0.498 mmol) and
TFA (2 mL) in CH.sub.2Cl.sub.2 (10 mL) was stirred at ambient
conditions for 3 hours. The volatile component was removed in vacuo
and the resulting material was loaded onto an MCX column, flushed
with methanol, eluted with 2.0 M NH.sub.3/methanol and concentrated
to provide
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)naphthalene as yellow solid (253 mg). LC-MS retention time
1.07 min; calcd. for C.sub.34H.sub.30N.sub.6: 522.25; found m/z
523.19 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM.Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.17 (s, 2H), 8.06 (d, J=8.2 Hz, 2H),
8.00 (s, 2H), 7.88 (d, J=8.6 Hz, 2H), 7.71-7.81 (m, 4H), 4.84 (dd,
J=10.8, 7.5 Hz, 2H), 3.47-3.53 (m, 2H), 2.80 (dd, J=12.8, 7.6 Hz,
2H), 2.54-2.63 (m, 2H), 2.04-2.11 (m, 2H), 1.20-1.27 (m, 2H), 1.02
(q, J=7.7 Hz, 2H).
##STR00355## ##STR00356##
##STR00357##
Mixture of Intermediate 30
(S)-Benzyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-b-
enzo[d]imidazol-6-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine--
1-carboxylate bistrifluoroacetate
And Intermediate 31
(S)-Benzyl
2-(5-(4-(7-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-b-
enzo[d]imidazol-6-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine--
1-carboxylate bistrifluoroacetate
[0681] To a mixture of a TFA salt of (S)-tert-butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate and a TFA salt of
(S)-tert-butyl
2-(6-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)pyrrolidine-1-carboxylate (190 mg),
(S)-benzyl
2-(5-(4-bromophenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(Patent applications: WO 2008/021928, WO 2008/021936 and WO
2008/021927) (213 mg, 0.499 mmol), in 1,2-dimethoxyethane (10.0 mL)
and water (2.0 mL) was added NaHCO.sub.3 (140 mg, 1.663 mmol). The
reaction mixture was degassed in vacuo for 5 minutes and was
flushed with nitrogen. Pd(Ph.sub.3P).sub.4 (19 mg, 0.017 mmol) was
added and the pressure flask was capped and heated with an oil bath
at 85.degree. C. for 16 hours. The volatile component was removed
in vacuo and the residue was partitioned between ethyl acetate and
water. The layers were separated and the aqueous phase was
extracted several times with ethyl acetate. The combined organic
phases were dried (Na.sub.2SO.sub.4), filtered and concentrated in
vacuo. The resulting material was purified with flash
chromatography (sample was dry loaded on silica gel and eluted with
0-5% methanol/CH.sub.2Cl.sub.2) to afford partially purified
products. The residue was further purified by a reverse phase HPLC
(water/acetonitrile/TFA) to provide a mixture of a TFA salt of
(S)-benzyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imi-
dazol-6-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxyl-
ate and a TFA salt of (S)-benzyl
2-(5-(4-(7-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imi-
dazol-6-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxyl-
ate (381 mg) which was used without further purification. LC-MS
retention time 1.54 min; calcd. for C.sub.47H.sub.46N.sub.6O.sub.4
758.36; found m/z 759.37 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA.
##STR00358##
Mixture of Intermediate 32
(S)-Benzyl
2-(5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)na-
phthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
And Intermediate 33
(S)-Benzyl
2-(5-(4-(7-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)na-
phthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0682] A mixture of a TFA salt of (S)-benzyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imi-
dazol-6-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxyl-
ate and a TFA salt of (S)-benzyl
2-(5-(4-(7-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-benzo[d]imi-
dazol-6-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxyl-
ate (381 mg) and TFA (2 mL) in CH.sub.2Cl.sub.2 (10 mL) was stirred
at ambient conditions for 4 hours. The volatile component was
removed in vacuo and the crude material was purified by a reverse
phase HPLC (0 to 50% water/acetonitrile/NH.sub.4OAc) to provide a
mixture of (S)-benzyl
2-(5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalen-2-
-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate and
(S)-benzyl
2-(5-(4-(7-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalen-2-
-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (70 mg, 21%
yield) as yellow oil. LC-MS retention time 2.87 min; calcd. for
C.sub.42H.sub.38N.sub.6O.sub.2 658.31; found m/z 659.47
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 80% Solvent A/20% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold time
of 1 min and an analysis time of 5 min where Solvent A was 5%
acetonitrile/95% water/10 mmol NH.sub.4OAc and Solvent B was 95%
acetonitrile/5% water/10 mmol NH.sub.4OAc. .sup.1H NMR (Mixture of
compounds, 500 MHz, MeOD) .delta. ppm 8.07-8.24 (m, 4H), 7.91-8.03
(m, 6H), 7.73-7.89 (m, 12H), 7.66-7.73 (m, 4H), 7.27-7.46 (m, 6H),
7.09-7.18 (m, 4H), 6.97-7.04 (m, 2H), 5.01-5.21 (m, 5H), 4.83-5.01
(m, 2H), 3.72-3.82 (m, 2H), 3.53-3.65 (m, 2H), 3.42-3.52 (m, 2H),
3.35-3.42 (m, 4H), 2.48-2.62 (m, 2H), 2.28-2.47 (m, 4H), 2.04-2.24
(m, 8H).
##STR00359##
Intermediate 34
2-((S)-Pyrrolidin-2-yl)-6-(6-(4-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)-
phenyl)naphthalen-2-yl)-1H-benzo[d]imidazole tetratrifluoroacetate
and
Intermediate 35
2-((S)-Pyrrolidin-2-yl)-6-(7-(4-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)-
phenyl)naphthalen-2-yl)-1H-benzo[d]imidazole
tetratrifluoroacetate
[0683] To a solution of a mixture of (S)-benzyl
2-(5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalen-2-
-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate and
(S)-benzyl
2-(5-(4-(7-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalen-2-
-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (70 mg.
0.057 mmol) and potassium carbonate (K.sub.2CO.sub.3) in methanol
(5 mL) was added a slurry of palladium on carbon (10%, 11 mg) in
methanol (2 mL) at ambient temperature. The reactor was vacuum
purged and backfilled with hydrogen. The reaction was stirred for
16 h under a balloon of hydrogen. The mixture was filtered through
a pad of diatomaceous earth (CELITE.RTM.) under vacuum and the
filtrate was evaporated in vacuo. The crude material was purified
by a reverse phase HPLC (0 to 50% water/acetonitrile/TFA) to
provide a TFA salt of
2-((S)-pyrrolidin-2-yl)-6-(6-(4-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl-
)phenyl)naphthalen-2-yl)-1H-benzo[d]imidazole (50 mg) as white
solid and a TFA salt of
2-((S)-pyrrolidin-2-yl)-6-(7-(4-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl-
)phenyl)naphthalen-2-yl)-1H-benzo[d]imidazole (24 mg) as white
solid.
[0684] Analytical data for Intermediate 34: LC-MS retention time
1.06 min; calcd. for C.sub.34H.sub.32N.sub.6 524.27; found m/z
525.26 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 80% Solvent A/20%
[0685] Solvent B to 0% Solvent A/100% Solvent B, a gradient time of
3 min, a hold time of 1 min and an analysis time of 4 min where
Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent B was
90% acetonitrile/10% water/0.1% TFA. .sup.1H NMR (TFA salt, 500
MHz, MeOD) .delta. ppm 8.19 (d, J=10.4 Hz, 2H), 8.02-8.10 (m, 3H),
7.84-7.96 (m, 7H), 7.77-7.83 (m, 2H), 5.14 (t, J=7.8 Hz, 1H),
5.06-5.11 (m, 1H), 3.52-3.67 (m, 4H), 2.61-2.74 (m, 2H), 2.43-2.56
(m, 2H), 2.30-2.42 (m, 2H), 2.18-2.30 (m, 2H).
[0686] Analytical data for Intermediate 35: LC-MS retention time
1.13 min; calcd. for C.sub.34H.sub.32N.sub.6 524.27; found m/z
525.24 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 80% Solvent A/20% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (TFA salt, 500 MHz,
MeOD) .delta. ppm 8.27 (d, J=7.0 Hz, 2H), 7.99-8.06 (m, 3H),
7.90-7.96 (m, 4H), 7.87 (td, J=8.2, 1.7 Hz, 2H), 7.76-7.85 (m, 3H),
5.13 (t, J=7.8 Hz, 1H), 5.03-5.08 (m, 1H), 3.51-3.67 (m, 4H),
2.61-2.74 (m, 2H), 2.42-2.54 (m, 2H), 2.20-2.40 (m, 4H).
##STR00360## ##STR00361##
##STR00362##
Intermediate 36
(S)-tert-Butyl
2-(6-bromo-1H-imidazo[4,5-b]pyridin-2-yl)pyrrolidine-1-carboxylate
[0687] HATU (3.64 g, 9.57 mmol) was added to a stirred solution of
5-bromopyridine-2,3-diamine (1.64 g, 8.70 mmol) and
(S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (1.87 g,
8.70 mmol) in DMF (50 mL) and DIPEA (3.19 mL, 18.3 mmol). The
reaction mixture was stirred at room temperature for 3 d, diluted
with water (400 mL) and extracted with EtOAc (150 mL). The organic
layer was washed with brine (50 mL), dried (MgSO.sub.4), filtered
and concentrated. The crude material was partially purified by
flash chromatography (110 g SiO.sub.2, 1-4% MeOH/DCM) to yield 3.36
g of solidified foam. The material was dissolved into AcOH (35.0
mL) and heated at 100.degree. C. for 8 h. The reaction was cooled
to room temperature, concentrated and purified by flash
chromatography (loaded with DCM, 80 g SiO.sub.2, 20-40%
EtOAc/Hexanes) to yield (S)-tert-butyl
2-(6-bromo-1H-imidazo[4,5-b]pyridin-2-yl)pyrrolidine-1-carboxylate
(1.73 g) as yellow solid. LC-MS retention time 1.33 min; m/z 365,
367 (1:1) (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.40 (br s, 1H), 8.09
(br s, 1H), 5.12-4.96 (m, 1H), 3.77-3.67 (m, 1H), 3.62-3.51 (m,
1H), 2.56-2.38 (m, 1H), 2.15-1.90 (m, 3H), 1.46 (s, 3.5H), 1.16 (s,
5.5H).
##STR00363##
Mixture of Intermediate 37
(S)-tert-Butyl
2-(6-bromo-3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin--
2-yl)pyrrolidine-1-carboxylate
And Intermediate 38
(S)-tert-Butyl
2-(6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyridin--
2-yl)pyrrolidine-1-carboxylate
[0688] A 60% dispersion of NaH (0.120 g, 3.00 mmol) was added to a
stirred solution of (S)-tert-butyl
2-(6-bromo-1H-imidazo[4,5-b]pyridin-2-yl)pyrrolidine-1-carboxylate
(1.0 g, 2.7 mmol) in DMF (25 mL) and the reaction was stirred for
1.5 h at room temperature. Then SEM-Cl (0.483 mL, 2.72 mmol) was
added and the reaction was stirred 16 h. The reaction was diluted
with water (.about.30 mL) and EtOAc (.about.35 mL), the layers were
separated and the organic layer was washed with brine (30 mL),
dried (MgSO.sub.4), filtered and concentrated. The residual crude
yellow oil was purified by flash chromatography (80 g SiO.sub.2,
20-30% EtOAc/hexanes) to yield a mixture of (S)-tert-butyl
2-(6-bromo-3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin--
2-yl)pyrrolidine-1-carboxylate and (S)-tert-butyl
2-(6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyridin--
2-yl)pyrrolidine-1-carboxylate (838 mg) as a yellow oil. The
mixture was used w/o further purification. LC-MS retention time
2.54 min; m/z 497,499 (1:1) (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 5 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% acetonitrile/95% water/10 mM
ammonium acetate and Solvent B was 5% water/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00364##
Intermediate 39
(2S,2'S)-tert-Butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(3-((2-(trimethylsilyl)ethoxy)methyl)-
-3H-imidazo[4,5-b]pyridine-6,2-diyl))dipyrrolidine-1-carboxylate
and
Intermediate 40
(S)-tert-Butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-3-(-
(2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyrrolidin-
e-1-carboxylate
[0689] In a microwave vial,
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene (57
mg, 0.150 mmol), a mixture of (S)-tert-butyl
2-(6-bromo-3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin--
2-yl)pyrrolidine-1-carboxylate and (5)-tert-butyl
2-(6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyridin--
2-yl)pyrrolidine-1-carboxylate (112 mg), cesium carbonate (147 mg,
0.450 mmol) and dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine
(12.3 mg, 0.030 mmol) were dissolved into THF (3 mL) and water (0.3
mL). An additional 1.5 mL of THF was added and the reaction was
sparged with bubbling nitrogen until .about.2 mL had evaporated
away. To the clear solution was added palladium (II) acetate (3.37
mg, 0.015 mmol). The vial was flushed with nitrogen, sealed and
then heated with microwave irradiation at 120.degree. C. for 30
min. The reaction was diluted with EtOAc (.about.3 mL) and washed
with water (2 mL) and brine (2 mL). The reaction was dried
(MgSO.sub.4), filtered and concentrated to a yellow oil which was
purified by flash chromatography (12 g SiO.sub.2, 25-75%
EtOAc/hexanes) to yield (2S,2'S)-tert-butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(3-((2-(trimethylsilyl)ethoxy)methyl)-
-3H-imidazo[4,5-b]pyridine-6,2-diyl))dipyrrolidine-1-carboxylate
(74 mg) as a clear colorless oil (mixture of SEM regioisomers) and
(S)-tert-butyl
2-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-3-(-
(2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyrrolidin-
e-1-carboxylate (32 mg) as a colorless oil (mixture of SEM
regioisomers). Each was used without further purification.
[0690] Analytical data for Intermediate 39: LC-MS retention time
3.42 min; m/z 961.9 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) (mixture of SEM regioisomers)
.delta. ppm 8.84-8.75 (m, 2H), 8.39-8.31 (m, 2H), 8.26-8.18 (m,
2H), 8.11 (br s, 2H), 7.92-7.84 (m, 2H), 5.94-5.83 (m, 3H), 5.79
(d, J=11.0 Hz, 1H), 5.34-5.25 (m, 2H), 3.86-3.57 (m, 8H), 2.61-2.39
(m, 2H), 2.34-1.99 (m, 6H), 1.45 (s, 8H), 1.17 (s, 10H), 1.10-0.87
(m, 4H), -0.02 (s, 18H).
[0691] Analytical data for Intermediate 40: LC-MS retention time
3.15 min; m/z 671.5 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. (mixture of SEM regioisomers) .delta. ppm 8.82-8.75 (m, 1H),
8.39-8.29 (m, 2H), 8.21-8.15 (m, 1H), 8.07-8.01 (m, 1H), 8.11 (br
s, 2H), 7.98-7.93 (m, 1H), 7.89-7.80 (m, 2H), 5.94-5.85 (m, 1.5H),
5.79 (d, J=11.3 Hz, 0.5H), 5.33-5.24 (m, 1H), 3.85-3.56 (m, 4H),
2.62-2.39 (m, 1H), 2.33-1.99 (m, 1H), 1.49-0.87 (m, 23H), -0.02 (s,
9H).
##STR00365##
Intermediate 41
2,6-Bis(2-((S)-pyrrolidin-2-yl)-3H-imidazo[4,5-b]pyridin-6-yl)naphthalene
[0692] 4M Hydrogen chloride (2.0 mL, 8.0 mmol) in dioxane was added
to a stirred solution of (2S,2'S)-tert-butyl
2,2'-(6,6'-(naphthalene-2,6-diyl)bis(3-((2-(trimethylsilyl)ethoxy)methyl)-
-3H-imidazo[4,5-b]pyridine-6,2-diyl))dipyrrolidine-1-carboxylate
(74 mg, 0.077 mmol) (mixture of SEM regioisomers) in methanol (1
mL). The reaction was stirred at room temperature for 16 h and then
concentrated to dryness to yield a hydrochloride salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-3H-imidazo[4,5-b]pyridin-6-yl)naphthalene
(51.1 mg) as yellow solid which was used without further
purification. LC-MS retention time 0.83 min; m/z 501.13 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Sunfire 5u C18 4.6.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 5 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 10% acetonitrile/90% water/0.1%
trifluoroacetic acid and Solvent B was 10% water/90%
acetonitrile/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, D.sub.2O) .delta. ppm 8.76 (s, 2H), 8.54 (s,
2H), 8.07-7.97 (m, 4H), 7.86-7.78 (m, 2H), 5.26-5.16 (m, 2H),
3.98-3.65 (m, 8H), 2.79-2.63 (m, 2H), 2.46-2.25 (m, 6H).
##STR00366## ##STR00367##
##STR00368##
Intermediate 42
(S)-tert-Butyl
2-(6-bromo-1H-imidazo[4,5-b]pyrazin-2-yl)pyrrolidine-1-carboxylate
[0693] HATU (5.05 g, 13.3 mmol) was added to a stirred solution of
5-bromopyrazine-2,3-diamine (2.28 g, 12.1 mmol) and
(S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (2.60 g,
12.0 mmol) in DMF (50 mL) and DIPEA (4.42 mL, 25.3 mmol). The
reaction mixture was stirred at room temperature for 3 d and then
at 90.degree. C. for 1 d. The reaction mixture was diluted with
water (350 mL) and extracted with EtOAc (200 mL). The organic layer
was washed with brine (100 mL), dried (MgSO.sub.4), filtered and
concentrated. The crude material was partially purified by flash
chromatography (110 g SiO.sub.2, 1-4% MeOH/DCM) to yield 1.9 g of
solidified foam. The material was dissolved into AcOH (35.0 mL) and
heated at 95.degree. C. for 8 h. The reaction was cooled to room
temperature, concentrated and purified by flash chromatography
(loaded with DCM, 80 g SiO.sub.2, 20-30% EtOAc/Hexanes) to yield
(S)-tert-butyl
2-(6-bromo-1H-imidazo[4,5-b]pyrazin-2-yl)pyrrolidine-1-carboxylate
(760 mg, 2.06 mmol, 17% yield) as yellow solidified foam. LC-MS
retention time 1.28 min; m/z 366, 368 (1:1) (MH-). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 5 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% acetonitrile/95% water/10 mM
ammonium acetate and Solvent B was 5% water/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.50-8.43 (m, 1H), 5.11-4.99 (m, 1H), 3.77-3.68 (m,
1H), 3.62-3.51 (m, 1H), 2.57-2.40 (m, 1H), 2.19-1.95 (m, 3H), 1.46
(s, 3.5H), 1.17 (s, 5.5H).
##STR00369##
Intermediate 43
(S)-tert-Butyl
2-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyrazin--
2-yl)pyrrolidine-1-carboxylate and
Intermediate 44
(S)-tert-Butyl
2-(6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyrazin--
2-yl)pyrrolidine-1-carboxylate
[0694] A 60% dispersion of NaH (75 mg, 1.9 mmol) was added to a
stirred solution of (S)-tert-butyl
2-(6-bromo-1H-imidazo[4,5-b]pyrazin-2-yl)pyrrolidine-1-carboxylate
(627 mg, 1.70 mmol) in DMF (15 mL) and the reaction was stirred for
1.5 h at room temperature. Then SEM-Cl (0.30 mL, 1.7 mmol) was
added and the reaction was stirred overnight. The reaction was
diluted with water (.about.30 mL) and EtOAc (.about.35 mL), the
layers were separated and the organic layer was washed with brine
(30 mL), dried (MgSO.sub.4), filtered and concentrated. The crude
yellow oil was purified by flash chromatography (40 g SiO.sub.2,
20-30% EtOAc/hexanes) to yield (S)-tert-butyl
2-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyrazin--
2-yl)pyrrolidine-1-carboxylate (421 mg) as a clear colorless oil
and (S)-tert-butyl
2-(6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyrazin--
2-yl)pyrrolidine-1-carboxylate (345 mg) as a clear colorless oil.
The absolute regiochemistry of the SEM group was not established
unambiguously, the names (and structures) may be exchanged in these
intermediates.
[0695] Analytical data for Intermediate 43: LC-MS retention time
2.43 min; m/z 496, 498 (1:1) (MH-). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 5 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% acetonitrile/95% water/10 mM
ammonium acetate and Solvent B was 5% water/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.52 (s, 0.5H), 8.49 (s, 0.5H), 5.90-5.72 (m, 2H),
5.32-5.25 (m, 1H), 3.82-3.54 (m 4H), 2.59-2.41 (m, 1H), 2.32-1.98
(m, 3H), 1.43 (s, 4.5H), 1.15 (s, 4.5H), 1.07-0.85 (m, 2H), -0.02
(s, 9H).
[0696] Analytical data for Intermediate 44: LC-MS retention time
2.41 min; m/z 496, 498 (1:1) (MH-). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 5 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% acetonitrile/95% water/10 mM
ammonium acetate and Solvent B was 5% water/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.61 (s, 0.5H), 8.58 (s, 0.5H), 5.88-5.71 (m, 2H), 5.28
(dd, J=8.5, 4.5 Hz, 1H), 3.82-3.54 (m 4H), 2.59-2.41 (m, 1H),
2.32-1.98 (m, 3H), 1.43 (s, 4.5H), 1.15 (s, 4.5H), 1.09-0.85 (m,
2H), -0.02 (s, 9H).
##STR00370##
Intermediate 45
(2S,2'S)-tert-Butyl
2,2'-(5,5'-(naphthalene-2,6-diyl)bis(1-((2-(trimethylsilyl)ethoxy)methyl)-
-1H-imidazo[4,5-b]pyrazine-5,2-diyl))dipyrrolidine-1-carboxylate
and
Intermediate 46
(S)-tert-Butyl
2-(5-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1-(-
(2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyrazin-2-yl)pyrrolidin-
e-1-carboxylate
[0697] In a microwave vial,
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(149 mg, 0.393 mmol), (S)-tert-butyl
2-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyrazin--
2-yl)pyrrolidine-1-carboxylate (294 mg, 0.590 mmol) (or a SEM
regioisomer), cesium carbonate (384 mg, 1.18 mmol) and
dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine (32.3 mg, 0.079
mmol) were dissolved into THF (4 mL) and water (0.4 mL). An
additional 1.5 mL of THF was added and the reaction was sparged
with bubbling nitrogen until the 1.5 mL had evaporated away. To the
clear solution was added palladium(II) acetate (8.83 mg, 0.039
mmol). The vial was flushed with nitrogen, sealed and then heated
with microwave irradiation at 120.degree. C. for 30 min. The
reaction was diluted with EtOAc (.about.3 mL) and washed with water
(2 mL) and brine (2 mL). The reaction was dried (MgSO.sub.4),
filtered and concentrated to a yellow oil which was purified by
flash chromatography (12 g SiO.sub.2, 25-75% EtOAc/hexanes) to
yield (2S,2'S)-tert-butyl
2,2'-(5,5'-(naphthalene-2,6-diyl)bis(1-((2-(trimethylsilyl)ethoxy)methyl)-
-1H-imidazo[4,5-b]pyrazine-5,2-diyl))dipyrrolidine-1-carboxylate
(or a SEM regioisomer) (210 mg) as a yellow solidified foam and
(S)-tert-butyl
2-(5-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1-(-
(2-(trimethylsilyl)ethoxy)methyl)-1H-imidazo[4,5-b]pyrazin-2-yl)pyrrolidin-
e-1-carboxylate (or a SEM regioisomer) (53.5 mg) as a colorless
oil.
[0698] Analytical date for Intermediate 45: LC-MS retention time
3.27 min; m/z 963.89 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) (mixture of SEM regioisomers)
.delta. ppm 9.29-9.15 (m 2H), 8.79-8.66 (m, 2H), 8.43-8.29 (m, 2H),
8.20-7.98 (m, 2H), 6.03-5.84 (m, 4H), 5.34 (dd, J=8.3, 4.3 Hz, 2H),
3.87-3.70 (m, 6H), 3.68-3.58 (m, 2H), 2.63-2.43 (m, 2H), 2.36-1.14
(m, 4H), 2.14-1.99 (m, 2H), 1.46 (s, 8H), 1.17 (s, 10H), 1.35-0.85
(m, 4H), -0.02 (s, 18H).
[0699] Analytical date for Intermediate 45: LC-MS retention time
3.07 min; m/z 672.52 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00371##
Intermediate 47
2,6-Bis(2-((S)-pyrrolidin-2-yl)-1H-imidazo[4,5-b]pyrazin-5-yl)naphthalene
[0700] 4M Hydrogen chloride (6.0 mL, 24 mmol) in dioxane was added
to a stirred solution of (2S,2'S)-tert-butyl
2,2'-(5,5'-(naphthalene-2,6-diyl)bis(1-((2-(trimethylsilyl)ethoxy)methyl)-
-1H-imidazo[4,5-b]pyrazine-5,2-diyl))dipyrrolidine-1-carboxylate
(210 mg, 0.218 mmol) (absolute regiochemistry of the SEM was not
established) in methanol (3 mL). The reaction was stirred at room
temperature for 16 h and concentrated to dryness to yield a
hydrochloride salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-imidazo[4,5-b]pyrazin-5-yl)naphthalene
(143 mg) as orange solid which was used without further
purification. LC-MS retention time 0.90 min; m/z 503.21 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Sunfire 5u C18 4.6.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 5 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 10% acetonitrile/90% water/0.1%
trifluoroacetic acid and Solvent B was 10% water/90%
acetonitrile/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, D.sub.2O) .delta. ppm 8.59 (s, 2H), 7.94 (s,
2H), 7.85 (d, J=8.5 Hz, 2H), 7.76 (d, J=8.5 Hz, 2H), 5.23-5.16 (m,
2H), 3.98-3.66 (m, 8H), 2.78-2.64 (m, 2H), 2.47-2.28 (m, 6H).
##STR00372##
##STR00373##
Intermediate 48
tert-Butyl
(2S)-2-(5-(6-(2-((2S)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-n-
aphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxylate
[0701] To a solution of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(885 mg, 1.78 mmol), DMAP (10.8 mg, 0.089 mmol) and triethylamine
(1.2 mL, 8.9 mmol) in DMF (30 mL) at room temp was added in one
portion di-tert-butyl dicarbonate (387 mg, 1.78 mmol). The reaction
mixture was stirred overnight at room temperature and purified on a
BIOTAGE.RTM. (dry loaded to a 80 g silica gel cartridge and eluted
with a gradient of 0 to 50% DCM in MeOH), to yield tert-butyl
(2S)-2-(5-(6-(2-((2S)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1-
H-benzimidazol-2-yl)-1-pyrrolidinecarboxylate (500 mg, 0.835 mmol,
47.1% yield) as orange solid. LC-MS retention time 1.207 min; m/z
599.25 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.27 (d, J=3.1 Hz, 1H), 8.23 (s, 1H),
8.04-8.17 (m, 4H), 8.03 (s, 1H), 7.88-7.97 (m, 3H), 7.75-7.83 (m,
2H), 5.28-5.35 (m, 1H), 5.11 (t, J=7.8 Hz, 1H), 3.75-3.83 (m, 1H),
3.60-3.72 (m, 2H), 3.51-3.59 (m, 1H), 2.60-2.74 (m, 2H), 2.40-2.50
(m, 1H), 2.21-2.38 (m, 3H), 2.10-2.20 (m, 2H), 1.52 (s, 4H), 1.25
(s, 4H).
##STR00374##
Intermediate 49
tert-Butyl
(2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-L-valyl)-2-pyrroli-
dinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidi-
necarboxylate
[0702] To a solution of tert-butyl
(2S)-2-(5-(6-(2-((2S)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1-
H-benzimidazol-2-yl)-1-pyrrolidinecarboxylate (400 mg, 0.668 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (176 mg, 1.00
mmol) and DIEA (0.583 mL, 3.34 mmol) in DMF (10 mL) was added HATU
(381 mg, 1.00 mmol). The reaction mixture was stirred 2 h at room
temperature and then purified by prep HPLC (using a Waters Sunfire
C18 column 30.times.150 mm 5u eluted with a gradient of 0 to 50%
ACN-Water+0.1% TFA) to yield a TFA salt of tert-butyl
(2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-L-valyl)-2-pyrrolidinyl)-1H--
benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxyl-
ate (445 mg) as yellow solid. LC-MS retention time 1.317 min; m/z
756.28 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00375##
Intermediate 50
Methyl
((1S)-2-methyl-1-(((2S)-2-(5-(6-(2-((2S)-2-pyrrolidinyl)-1H-benzimi-
dazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)prop-
yl)carbamate
[0703] To a solution of a TFA salt of tert-butyl
(2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-L-valyl)-2-pyrrolidinyl)-1H--
benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinecarboxyl-
ate (445 mg) in DCM (10 mL) at room temp was added TFA (1 mL, 13
mmol). The mixture was stirred at rt for 3 h, concentrated, and the
residue was diluted in MeOH, loaded onto a Strata XC MCX cartridge
and washed with methanol. The compound was release from the
cartridge by eluting with a solution of 2M ammonia in methanol and
then evaporated under reduced pressure to give methyl
((1S)-2-methyl-1-(((2S)-2-(5-(6-(2-((2S)-2-pyrrolidinyl)-1H-benzimidazol--
5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)car-
bamate (275 mg) as orange solid LC-MS retention time 1.120 min; m/z
656.27 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.26 (br s, 1H), 8.22 (br s, 1H),
8.08-8.16 (m, 3H), 8.04-8.08 (m, 1H), 8.03 (s, 1H), 7.86-7.96 (m,
3H), 7.73-7.82 (m, 2H), 5.88-5.93 (m, 0.1H), 5.37-5.44 (m, 0.9H),
5.07-5.15 (m, 1H), 4.28-4.33 (m, 1H), 4.12-4.19 (m, 0.8H),
4.06-4.10 (m, 0.2H), 3.94-4.02 (m, 1H), 3.59-3.75 (m, 4H),
3.50-3.59 (m, 1H), 2.63-2.74 (m, 2H), 2.41-2.47 (m, 1H), 2.23-2.40
(m, 5H), 2.06-2.16 (m, 1H), 1.00-1.04 (m, 0.4H), 0.94-1.00 (m, 3H),
0.88-0.93 (m, 2.6H).
##STR00376## ##STR00377##
##STR00378##
Intermediate 51
(S)-tert-Butyl
2-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
imidazol-2-yl)pyrrolidine-1-carboxylate and
Intermediate 52
(2S,2'S)-tert-Butyl
2,2'-(4,4'-(naphthalene-2,6-diyl)bis(1H-imidazole-4,2-diyl))dipyrrolidine-
-1-carboxylate
[0704] A 100 mL pressure vessel equipped with a magnetic stir bar
was charged with
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(2.00 g, 5.26 mmol), (S)-tert-butyl
2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (2.10 g, 5.79
mmol) and Pd(Ph.sub.3P).sub.4 (0.058 g, 0.05 mmol) in DME (47.8 mL)
and water (4.8 mL). The solution was degassed under vacuum for 5
min and the reactor was back filled with nitrogen. The vessel was
sealed and the reaction mixture was heated overnight at 120.degree.
C. The reaction was cooled to room temperature and the volatiles
were removed under. The residue was partitioned between water and
EtOAc and the water layer was extracted with additional EtOAc. The
combined organic layers were dried (Na.sub.2SO.sub.4), filtered,
and concentrated. The crude product was purified with a
BIOTAGE.RTM. (dry loaded onto a 90 g silica gel cartridge and
eluted with a gradient of 0 to 30% EtOAc in DCM) and repurified
with a BIOTAGE.RTM. (dry loaded onto a 80 g silica gel cartridge
and eluted with a gradient of 0 to 50% EtOAc in DCM) to yield
(S)-tert-butyl
2-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
imidazol-2-yl)pyrrolidine-1-carboxylate (949 mg) as a yellow solid.
The material (contaminated with (2S,2'S)-tert-butyl
2,2'-(4,4'-(naphthalene-2,6-diyl)bis(1H-imidazole-4,2-diyl))dipyrrolidine-
-1-carboxylate) was used in the next step without further
purification. LC-MS retention time 1.760 min; m/z 490.21
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode.
[0705] The preparative HPLC from the next reaction yielded
(2S,2'S)-tert-butyl
2,2'-(4,4'-(naphthalene-2,6-diyl)bis(1H-imidazole-4,2-diyl))dipyrrolidine-
-1-carboxylate (92 mg, 0.111 mmol, 11% yield) as yellow solid LC-MS
retention time 1.113 min; m/z 599.24 [M+H].sup.+. LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Waters Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.35 (br s, 2H), 8.13 (br
s, 2H), 8.08 (br s, 1H), 8.00 (br s, 1H), 7.95 (m, 2H), 5.18 (br s,
2H), 3.68-3.78 (m, 2H), 3.62 (br s, 2H), 2.51-2.68 (m, 2H), 2.21
(br s, 2H), 2.06-2.15 (m, 4H), 1.50 (br s, 9H), 1.31 (br s,
9H).
##STR00379##
Intermediate 53
(S)-tert-Butyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-4--
yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0706] A 100 mL pressure vessel equipped with a magnetic stir bar
was charged with (S)-tert-butyl
2-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
imidazol-2-yl)pyrrolidine-1-carboxylate (500 mg, 1.02 mmol),
(S)-tert-butyl
2-(5-(4-bromophenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(601 mg, 1.53 mmol) and Pd(Ph.sub.3P).sub.4 (58 mg, 0.05 mmol) in
DME (10 mL) and water (1.0 mL). The solution was degassed under
vacuum for 5 min and the reactor was back filled with nitrogen. The
mixture was heated for 8 h at 120.degree. C., additional
Pd(Ph.sub.3P).sub.4 (57.8 mg, 0.05 mmol) was added and the mixture
was degassed and stirred overnight at 130.degree. C. The reaction
was cooled to room temperature and the volatiles were removed under
vacuum. The residue was partitioned between water and EtOAc and the
water layer was extracted with additional EtOAc. The combined
organic layers were dried (Na.sub.2SO.sub.4), filtered, and
concentrated. The crude product was purified on a BIOTAGE.RTM. (dry
loaded onto a 90 g silica gel cartridge and eluted with a gradient
of 5 to 100% EtOAc in DCM) and then repurified by prep HPLC (using
a Waters Sunfire C18 column 30.times.100 mm 5u eluted
MeOH-Water+0.1% TFA) to yield a TFA salt of (S)-tert-butyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-4--
yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(50 mg). LC-MS retention time 1.278 min; m/z 675.70 [M+H].sup.+. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Waters Sunfire 5u C18 4.6.times.50 mm column using
a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min and an analysis time
of 4 min where Solvent A was 10% acetonitrile/90% water/0.1% TFA
and Solvent B was 90% acetonitrile/10% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.31 (d, J=15.9 Hz,
2H), 8.14-8.18 (m, 1H), 8.10 (d, J=8.85 Hz, 1H), 8.05 (s, 0.6H),
7.96-8.03 (m, 4H), 7.85-7.94 (m, 3.4H), 5.11-5.23 (m, 2H),
3.69-3.77 (m, 2H), 3.62 (br s, 2H), 2.51-2.67 (m, 2H), 2.16-2.26
(m, 2H), 2.05-2.16 (m, 4H), 1.51 (s, 9H), 1.32 (s, 9H).
##STR00380##
Intermediate 54
2-((2S)-2-Pyrrolidinyl)-4-(4-(6-(2-((2S)-2-pyrrolidinyl)-1H-imidazol-4-yl)-
-2-naphthyl)phenyl)-1H-imidazole
[0707] To a solution of a TFA salt of (S)-tert-butyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-4--
yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(50 mg) in DCM (10 mL) was added TFA (2.0 mL, 26 mmol) in one
portion. The mixture was agitated at room temperature for 2 h. The
reaction was concentrated under vacuum to yield a TFA salt of
2-((S)-pyrrolidin-2-yl)-5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl-
)naphthalen-2-yl)phenyl)-1H-imidazole (51.5 mg) as orange solid,
which was used without further purification. LC-MS retention time
0.978 min; m/z 475.19 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
HPLC-MS (TFA) 78457-087 using a Waters Sunfire C18 column 4
6.times.50 mm 5u showed the reaction was complete peak found at
RT=0.978 min. MH.sup.+=475.19. .sup.1H NMR (500 MHz, MeOD) .delta.
ppm 8.34 (s, 1H), 8.20 (s, 1H), 8.03 (dd, J=14.2, 8.7 Hz, 2H),
7.84-7.96 (m, 7H), 7.79-7.84 (m, 1H) 4.99-5.11 (m, 2H), 3.48-3.63
(m, 4H), 2.57-2.70 (m, 2H), 2.41-2.55 (m, 2H), 2.29-2.41 (m, 2H),
2.15-2.29 (m, 2H).
##STR00381##
Intermediate 55
4,4'-(2,6-Naphthalenediyl)bis(2-(2S)-2-pyrrolidinyl)-1H-imidazole)
[0708] To a solution of a TFA salt of (2S,2'S)-tert-butyl
2,2'-(4,4'-(naphthalene-2,6-diyl)bis(1H-imidazole-4,2-diyl))dipyrrolidine-
-1-carboxylate (82 mg) in DCM (2 mL) at room temperature was added
TFA (2.0 mL, 26 mmol) in one portion. The mixture was stirred for 3
h at room temperature, purified by prep HPLC (Waters Sunfire C18
column 30.times.100 mm 5u eluted with a gradient of 5 to 90%
MeOH-Water+0.1% TFA) and repurified by prep HPLC (PHENOMENEX.RTM.
Luna C18 column 30.times.100 mm 10u eluted with a gradient of 10 to
90% methanol-Water+0.1% TFA) to yield a TFA salt of
4,4'-(2,6-naphthalenediyl)bis(2-((2S)-2-pyrrolidinyl)-1H-imidazole)
(68 mg) as tan solid. LC-MS retention time 0.978 min; m/z 399.18
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.31
(s, 2H), 7.95-8.00 (m, 2H), 7.91-7.95 (m, 2H), 7.85 (s, 2H), 5.03
(t, J=8.2 Hz, 2H), 3.49-3.62 (m, 4H), 2.59-2.67 (m, 2H), 2.42-2.52
(m, 2H), 2.31-2.40 (m, 2H), 2.17-2.28 (m, 2H).
##STR00382##
##STR00383##
Intermediate 56
(S)-tert-Butyl
2-(5-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0709] (S)-tert-Butyl
2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (750 mg, 2.07
mmol) and copper(I) iodide (79 mg, 0.41 mmol) were dissolved into
DMF (20 mL) and triethylamine (1.439 mL, 10.33 mmol). The reaction
solution was vacuum flushed for 10 minutes (aspirator vacuum,
flushed with nitrogen) and then ethynyltrimethylsilane (1.2 mL, 8.3
mmol) and finally Pd(PPh.sub.3).sub.4 (119 mg, 0.103 mmol) were
added. The flask was flushed with nitrogen, seal and heated at
50.degree. C. overnight. The reaction was concentrated to a crude
black tar, dissolved into DCM and purified via BIOTAGE.RTM. Horizon
(80 g SiO.sub.2, 20-40% EtOAc/hexanes) to yield (S)-tert-butyl
2-(5-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(546 mg, 1.64 mmol, 79% yield) as a yellow solid. This material was
used without further purification. LC-MS retention time 1.393 min;
m/z 332.33 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and Solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.19-7.13 (m,
1H), 4.92-4.85 (m, 1H), 3.43-3.31 (m, 2H), 3.04-2.83 (m, 1H),
2.20-1.88 (m, 3H), 1.51 (s, 3H), 1.49 (s, 6H), 0.25 (s, 3H), 0.23
(s, 6H).
##STR00384##
Intermediate 57
(S)-tert-Butyl
2-(5-ethynyl-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0710] (S)-tert-Butyl
2-(5-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(537 mg, 1.61 mmol) was dissolved into MeOH (20 mL) and then
potassium carbonate (22 mg, 0.16 mmol) was added and the reaction
was stirred at rt for 3 h. The reaction was concentrated, dissolved
into dichchloromethane, loaded onto a SiO.sub.2 column and purified
by BIOTAGE.RTM. Horizon (30-50% EtOAc/hexanes) to yield
(S)-tert-butyl
2-(5-ethynyl-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (346 mg,
1.32 mmol, 82% yield) as an off-white solid. LC-MS retention time
0.878 min; m/z 260.35 (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and Solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.21-7.17 (m,
1H), 4.92-4.87 (m, 1H), 3.43-3.30 (m, 2H), 3.05 (s, 1H), 3.03-2.85
(m, 1H), 2.20-1.88 (m, 3H), 1.60-1.45 (m, 9H).
##STR00385##
Intermediate 58
tert-Butyl
(2S)-2-(4-((6-((2-((2S)-1-(tert-butoxycarbonyl)-2-pyrrolidinyl)-
-1H-imidazol-5-yl)ethynyl)-2-naphthyl)ethynyl)-1H-imidazol-2-yl)-1-pyrroli-
dinecarboxylate
[0711] (S)-tert-Butyl
2-(5-ethynyl-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (340 mg,
1.30 mmol), 2,6-dibromonaphthalene (744 mg, 2.60 mmol) and
copper(I) iodide (12.39 mg, 0.065 mmol) were slurried into DMF (10
mL) and triethylamine (0.54 mL, 3.9 mmol). The solution was vacuum
flushed with nitrogen (5.times.), treated with Pd(PPh.sub.3).sub.4
(75 mg, 0.065 mmol) and then vacuum flushed with nitrogen
(2.times.). The cloudy yellow solution was stirred under nitrogen
at rt for 3 h. The reaction was concentrated to a yellow-orange
semi-solid, slurried into DMF/MeOH (.about.1:1), filtered and
purified in five injections by prep HPLC (acetonitrile/water with
10 mM ammonium acetate, 15-100%). The fractions containing the
desired product were combined and concentrated to yield tert-butyl
(2S)-2-(4-((6-((2-((2S)-1-(tert-butoxycarbonyl)-2-pyrrolidinyl)-1H-imidaz-
ol-5-yl)ethynyl)-2-naphthyl)ethynyl)-1H-imidazol-2-yl)-1-pyrrolidinecarbox-
ylate (70 mg, 0.11 mmol, 18% yield) as a light yellow solid. LC-MS
retention time 1.448 min; m/z 647.45 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 5 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time
of 3 min where Solvent A was 5% acetonitrile/95% H.sub.2O/10 mM
ammonium acetate and Solvent B was 5% H.sub.2O/95% acetonitrile/10
mM ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 10.70 (br s, 2H), 7.98 (s, 2H), 7.74 (d,
J=8.3 Hz, 2H), 7.57 (d, J=8.3 Hz, 2H), 7.30-7.25 (m, 2H, under
solvent peak), 4.95 (dd, J=7.3, 3.3 Hz, 2H), 3.46-3.37 (m, 4H),
3.08-2.91 (m, 2H), 2.24-2.08 (m, 4H), 2.03-1.92 (m, 2H), 1.51 (s,
18H).
##STR00386##
Intermediate 59
4,4'-(2,6-Naphthalenediyldi-2,1-ethynediyl)bis(2-(2S)-2-pyrrolidinyl)-1H-i-
midazole)
[0712] (2S,2'S)-tert-Butyl
2,2'-(5,5'-(naphthalene-2,6-diylbis(ethyne-2,1-diyl))bis(1H-imidazole-5,2-
-diyl))dipyrrolidine-1-carboxylate (32 mg, 0.049 mmol) was
dissolved into dichloroethane (2 mL) and then TFA (1 mL, 13 mmol)
was added. The solution was stirred at rt for 1 h. The reaction was
concentrated to dryness to yield a TFA salt of
4,4'-(2,6-naphthalenediyldi-2,1-ethynediyl)bis(2-(2S)-2-pyrrolidinyl)-1H--
imidazole) (41.6 mg) as tan solid. .sup.1H NMR (500 MHz,
CD.sub.3OD) .delta. ppm 8.05 (s, 2H), 7.89 (d, J=8.3 Hz, 2H), 7.59
(d, J=8.6 Hz, 2H), 7.52 (s, 2H), 4.89-4.84 (m, 2H), 3.56-3.44 (m,
4H), 2.59-2.50 (m, 2H), 2.42-2.24 (m, 4H), 2.26-2.14 (m, 2H).
##STR00387##
##STR00388##
Intermediate 60
(1R,5R)-tert-Butyl
3-(5-(4-bromophenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate
[0713] Hunig's Base (7.69 mL, 44.0 mmol) was added to a stirred
solution of 2-bromo-1-(4-bromophenyl)ethanone (12.23 g, 44.0 mmol)
and
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (10 g, 44 mmol) in dry MeCN (400 mL). The mixture was stirred
for 16 h at RT. The solvent was removed in vacuo and the residue
was taken up in ethyl acetate and washed with saturated sodium
bicarbonate solution and brine, dried (sodium sulfate), filtered
and concentrated. In a pressure vessel the residue was taken up in
xylene (400 mL) and ammonium acetate (33.9 g, 440 mmol) was added.
The vessel was sealed and heated at 140.degree. C. for 2 h. The
solvent was removed in vacuo and the residue was taken up in ethyl
acetate and washed with saturated sodium bicarbonate solution
(pH=9), brine, dried over sodium sulfate and concentrated. The
crude product, as a reddish orange foam, was dissolved in methylene
chloride and placed onto a 300 g Thompson silica gel cartridge
(eluted with 20% B to 100% B for 4000 mL where Solvent B=ethyl
acetate and Solvent A=hexanes) to yield (1R,3S,5R)-tert-butyl
3-(5-(4-bromophenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate (16.67 g, 39.1 mmol, 88.8% yield) as a golden-brown foam.
LC-MS retention time 1.762 min; m/z 403.94 [M+H].sup.+. LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna S10 3.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min and an analysis time of 5 min
where Solvent A was 5% methanol/95% water/0.1% TFA and Solvent B
was 95% methanol/5% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 7.62 (br d, J=8.6 Hz, 2H), 7.50 (br d,
J=8.6 Hz, 2H), 7.37 (s, 1H), 4.66 (br s, 1H), 3.58 (br s, 1H),
2.56-2.47 (m, 1H), 2.36-2.27 (m, 1H), 1.75-1.67 (m, 1H), 0.97 (br
s, 9H), 0.88-0.81 (m, 1H), 0.64-0.57 (m, 1H).
##STR00389##
##STR00390##
Intermediate 61
(1R,3S,5R)-tert-Butyl
3-(hydroxymethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0714] To a solution of
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (9.85 g, 43.3 mmol) in THF (200 mL) at 0.degree. C. was added
dropwise borane-methyl sulfide complex (282 mL, 563 mmol) over 30
min. The ice bath was removed, the mixture was stirred for 1 h and
then heated at reflux for 2 h. The mixture was cooled to 0.degree.
C., slowly quenched with methanol (.about.200 mL) and concentrated
under vacuum. The residue was dissolved in DCM and washed with
water (emulsion), 1N HCl, sat NaHCO.sub.3 aq, and brine. The
organic layer was dried over Na.sub.2SO.sub.4, filtered and
concentrated to yield (1R,3S,5R)-tert-butyl
3-(hydroxymethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (8.43 g,
39.5 mmol, 91% yield) as colorless oil. LC-MS retention time 1.398
min; m/z 236.20 [M+Na].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
methanol/90% water/0.1% TFA and Solvent B was 90% methanol/10%
water/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 3.72-3.79 (m, 1H), 3.52-3.64 (m, 3H), 3.15-3.24 (m,
1H), 2.00-2.08 (m, 1H), 1.62-1.72 (m, 1H), 1.54-1.62 (m, 1H),
1.45-1.51 (m, 9H), 0.84 (br s, 1H), 0.36 (td, J=5.0, 2.4 Hz,
1H).
##STR00391##
Intermediate 62
(1R,3S,5R)-tert-Butyl
3-formyl-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0715] To a solution of (1R,3S,5R)-tert-butyl
3-(hydroxymethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (4.85 g,
22.74 mmol) in DCM (125 mL) at 0.degree. C. was added Dess-Martin
periodinane (11.57 g, 27.3 mmol). The reaction was warmed up to RT
and stirred for 3 h. The reaction was poured into a saturated aq
solution of NaHCO.sub.3 then ca .about.20 g of
Na.sub.2S.sub.2O.sub.3 was added and stirred for 1 h. The layers
were separated and the aqueous layer was extracted several times
with DCM. The combined organic extracts were dried over
Na.sub.2SO.sub.4 and evaporated in vacuo. The residue was purified
by flash column chromatography (BIOTAGE.RTM.), eluting with a
gradient of 0 to 40% EtOAc/hexanes to afford (1R,3S,5R)-tert-butyl
3-formyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (4.47 g, 21.2
mmol, 93% yield) as colorless oil. LC-MS retention time 0.813 min;
m/z no ionization (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz,
chloroform-d) .delta. ppm 9.48 (br s, 1H), 3.67-3.97 (m, 1H),
3.33-3.59 (m, 1H), 2.10-2.31 (m, 2H), 1.56-1.67 (m, 1H), 1.47 (br
s, 9H), 0.80 (br s, 1H), 0.55 (br s, 1H).
##STR00392##
Intermediate 63
(1R,3S,5R)-tert-Butyl
3-(1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0716] Ammonium hydroxide (16.40 mL, 421 mmol) was added dropwise
to a stirred solution of (1R,3S,5R)-tert-butyl
3-formyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (4.4 g) in
methanol (15 mL) at 0.degree. C. The reaction was allowed to warm
to room temp and stirred for 45 min. Then, oxalaldehyde 40%
H.sub.2O (5.32 mL, 46.3 mmol) was added dropwise and the reaction
mixture was stirred overnight. The reaction color turned brown over
time. The reaction was diluted with brine and concentrated to
remove the volatile solvent. The remaining aqueous layer was then
extracted several times with ethyl acetate. The combined organic
layers were dried over Na.sub.2SO.sub.4 evaporated under vacuum.
The residue was purified by flash column chromatography
(BIOTAGE.RTM.), eluting with a gradient of 0 to 70% EtOAc/hexanes
to afford (1R,3S,5R)-tert-butyl
3-(1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (3.71
g, 14.9 mmol, 71% yield) as white solid. LC-MS retention time 0.772
min; m/z 250.20 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz,
chloroform-d) .delta. ppm 10.43 (br s, 1H), 6.99 (s, 2H), 4.86 (dd,
J=9.0, 5.0 Hz, 1H), 3.35 (br s, 1H), 3.20 (br s, 1H), 2.28-2.38 (m,
1H), 1.71-1.80 (m, 1H), 1.49 (s, 9H), 0.83-0.90 (m, 1H), 0.45 (br
s, 1H).
##STR00393##
Intermediate 64
(1R,3S,5R)-tert-Butyl
3-(4,5-diiodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0717] Iodine (I.sub.2, 12.43 g, 49.0 mmol) was added to a solution
of 3-(1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(5.55 g, 22.3 mmol) and sodium carbonate (7.08 g, 66.8 mmol) in
dioxane (56 mL) and water (56 mL) and the reaction mixture was
stirred in the dark for 16 h at room temperature. The reaction
mixture was diluted with EtOAc and washed an aqueous saturated
solution of sodium thiosulfate. The layers were separated and the
aqueous layer was extracted with EtOAc. The combined organic layers
were dried over Na.sub.2SO.sub.4 and evaporated under vacuum. The
residue was purified by flash column chromatography (BIOTAGE.RTM.),
eluting with a gradient of 0 to 100% EtOAc/hexanes to afford
(1R,3S,5R)-tert-butyl
3-(4,5-diiodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(8.3 g, 16 mmol, 71% yield) as white solid. LC-MS retention time
1.455 min; m/z 502.07 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 4.51 (br s, 1H), 3.56 (br s, 1H), 2.40-2.54 (m, 1H),
2.18-2.27 (m, 1H), 1.65-1.73 (m, 1H), 1.20-1.40 (br s, 9H), 0.82
(dt, J=8.6, 5.8 Hz, 1H), 0.58 (br s, 1H).
##STR00394##
Intermediate 65
(1R,3S,5R)-tert-Butyl
3-(4-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0718] A 250 mL pressure flask equipped with a stir bar was charge
with a solution of (1R,3S,5R)-tert-butyl
3-(4,5-diiodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(8.3 g, 17 mmol) in EtOH (83 mL) and water (83 mL). Then, sodium
sulfite (20.88 g, 166 mmol) was added and the mixture was heated at
90.degree. C. for 16 h. The reaction mixture was extracted with
EtOAc and the combined organic layers were dried over
Na.sub.2SO.sub.4 and evaporated under vacuum. The residue was
purified by flash column chromatography (BIOTAGE.RTM.), eluting
with a gradient of 0 to 15% EtOAc/DCM to afford
(1R,3S,5R)-tert-butyl
3-(4-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(3.84 g, 10.2 mmol, 62% yield) as yellow solid along with the fully
reduced product (1R,3S,5R)-tert-butyl
3-(1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (550
mg, 2.21 mmol, 13% yield). LC-MS retention time 0.932 min; m/z
376.22 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.12 (br
s, 1H), 4.59 (br s, 1H), 3.44-3.64 (br s, 1H), 2.40-2.52 (m, 1H),
2.17-2.30 (m, 1H), 1.65-1.73 (m, 1H), 1.27 (br s, 9H), 0.84 (dt,
J=8.5, 5.7 Hz, 1H), 0.58 (br s, 1H).
##STR00395##
##STR00396##
Intermediate 66
(1R,3S,5R)-tert-Butyl
3-(5-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)p-
henyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
and
Intermediate 67
(1R,1'R,3S,3'S,5R,5'R)-tert-Butyl
3,3'-(5,5'-(4,4'-(naphthalene-2,6-diyl)bis(4,1-phenylene))bis(1H-imidazol-
e-5,2-diyl))bis(2-azabicyclo[3.1.0]hexane-2-carboxylate)
[0719] A solution of
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(1.0 g, 2.6 mmol), (1R,3S,5R)-tert-butyl
3-(5-(4-bromophenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate (1.06 g, 2.63 mmol), Na.sub.2CO.sub.3 (0.837 g, 7.89 mmol) in
DME (20 mL) and water (2 mL) was degassed under vacuum for 10 min.
The mixture was heated at 80.degree. C. and then
Pd(Ph.sub.3P).sub.4 (0.152 g, 0.132 mmol) was added under a stream
of nitrogen. The reactor was sealed and the heating was pursued
further at 120.degree. C. for 16 h. The DME was removed in vacuo
and the crude material was partitioned between EtOAc/H.sub.2O. The
layers were separated and the aqueous layer was extracted several
times with EtOAc. The combined organic extracts were dried over
Na.sub.2SO.sub.4 and evaporated in vacuo. The residue was purified
by flash column chromatography (BIOTAGE.RTM.), eluting with a
gradient of 0 to 5% MeOH/DCM to afford the partially pure target
product contaminated with reaction side products. The impure
product was purified again by flash column chromatography
(BIOTAGE.RTM.), eluting with a gradient of 50 to 100%
EtOAc/hexanes, then the column was flushed with 10% MeOH/DCM to
afford (1R,3S,5R)-tert-butyl
3-(5-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)p-
henyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(606 mg) as yellow foam. LC-MS retention time 1.608 min; m/z 578.4
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent
[0720] A was 10% MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, chloroform-d, Partial NMR) .delta. ppm 4.89 (br s, 1H),
3.20-3.66 (m, 1H), 2.33-2.50 (m, 1H), 1.76-1.86 (m, 1H), 1.52 (br
s, 9H), 1.24-1.32 (m, 12H), 0.87-0.93 (m, 1H), 0.51 (br s, 1H).
##STR00397##
Intermediate 68
(1R,3S,5R)-tert-Butyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate
[0721] A solution of (1R,3S,5R)-tert-butyl
3-(5-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)p-
henyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(568 mg), (1R,3S,5R)-tert-butyl
3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(369 mg, 0.983 mmol), Na.sub.2CO.sub.3 (313 mg, 2.95 mmol) in DME
(10 mL) and water (2 mL) was degassed under vacuum for 10 min. The
mixture was heated at 80.degree. C. and then Pd(Ph.sub.3P).sub.4
(114 mg, 0.098 mmol) was added under a stream of nitrogen. The
reactor was sealed and the heating was pursued further at
130.degree. C. overnight. The DME was removed in vacuo and the
crude material was partitioned between EtOAc/H.sub.2O. The layers
were separated and the aqueous layer was extracted several times
with EtOAc. The combined organic extracts were dried over
Na.sub.2SO.sub.4 and evaporated in vacuo. The residue was purified
by flash column chromatography (BIOTAGE.RTM.), eluting with a
gradient of 20 to 100% EtOAc/hexanes, then 5% MeOH/DCM to afford
the partially pure (1R,3S,5R)-tert-butyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (277 mg). LC-MS retention time
1.578 min; m/z 699.56 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 7.62-7.69 (m, 6H), 7.53-7.59 (m, 4H), 6.95 (s, 2H),
4.67 (br s, 2H), 3.46-3.56 (m, 2H), 2.50-2.59 (m, 0.5H), 2.32-2.49
(m, 2H), 2.27 (br s, 1.5H), 1.65-1.78 (m, 2H), 1.28 (br. s, 18H),
0.80-0.88 (m, 2H), 0.53-0.66 (m, 2H).
##STR00398##
Intermediate 69
(1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hex-3-yl)-1H-imida-
zol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
[0722] To a solution of (1R,3S,5R)-tert-butyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (275 mg) in DCM (10 mL) was added
TFA (2 mL, 26.0 mmol). The mixture was stirred for 2 h at room
temperature. The volatiles were removed under vacuum and the crude
residue was purified by a reverse phase HPLC (water/MeOH/TFA) to
afford a TFA salt of
(1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imid-
azol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(106 mg) as tan solid. LC-MS retention time 1.153 min; m/z 499.36
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.33 (s,
1H), 8.19 (s, 1H), 7.99-8.05 (m, 2H), 7.85-7.95 (m, 6H), 7.83 (s,
1H), 7.76 (s, 1H), 4.78 (q, J=9.4 Hz, 2H), 3.47-3.54 (m, 2H),
2.65-2.74 (m, 4H), 2.02-2.12 (m, 2H), 1.18-1.25 (m, 2H), 0.97-1.05
(m, 2H).
##STR00399##
##STR00400##
Intermediate 70
tert-Butyl
(1R,3S,5R)-3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-6-quinolinyl)-1H-benzimidazo-
l-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0723] To a degassed solution of 6-bromo-2-chloroquinoline (30 mg,
0.12 mmol), Na.sub.2CO.sub.3 (39.3 mg, 0.371 mmol) and
(1R,3S,5R)-tert-butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (116 mg, 0.272 mmol) in
dioxane (1 mL) and H.sub.2O (0.2 mL) was added Pd(Ph.sub.3P).sub.4
(14.3 mg, 0.012 mmol) and the mixture was stirred at 110.degree. C.
for 2 h. The reaction mixture was diluted with MeOH, and purified
by prep HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield a TFA
salt of tert-butyl
(1R,3S,5R)-3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-6-yl)-6-quinolinyl)-1H-benzimidazol-2-yl)-2--
azabicyclo[3.1.0]hexane-2-carboxylate (60.6 mg, 0.057 mmol, 46.3%
yield) as a yellow solid. LC-MS retention time 2.05 min; m/z 724
[M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.68 (1H, d, J=8.5 Hz), 8.60 (1H, s), 8.44 (1H, dd,
J=8.5, 1.5 Hz), 8.39 (1H, d, J=1.3 Hz), 8.33 (1H, d, J=8.8 Hz),
8.23-8.29 (1H, m), 8.25 (1H, d, J=8.5 Hz), 8.19 (1H, s), 8.08 (1H,
d, J=9.0 Hz), 7.96 (1H, d, J=9.0 Hz), 7.93 (1H, d, J=9.5 Hz),
5.02-5.12 (2H, m), 3.70 (2H, br s), 2.74-2.85 (2H, m), 2.45-2.57
(2H, m), 1.84-1.96 (2H, m), 1.05-1.68 (18H, m), 0.89-1.01 (2H, m),
0.77-0.86 (2H, m).
##STR00401##
Intermediate 71
2,6-Bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-
-yl)quinoline
[0724] A TFA salt of tert-butyl
(1R,3S,5R)-3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-6-yl)-6-quinolinyl)-1H-benzimidazol-2-yl)-2--
azabicyclo[3.1.0]hexane-2-carboxylate (51 mg, 0.054 mmol) was mixed
with a dioxane solution of HCl (0.5 mL, 2.00 mmol), and stirred at
rt for 2 h. The volatiles were removed to yield an HCl salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)quinoline (53 mg) as yellow solid. LC-MS retention time 1.20
min; m/z 524 [M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50
mm S10. Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10%
water:90% methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0.
Final % B=100. Gradient Time=3 min. Wavelength=220).
##STR00402##
##STR00403##
Intermediate 72
tert-Butyl
(1R,3S,5R)-3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-6-quinoxalinyl)-1H-benzimida-
zol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0725] Pd(Ph.sub.3P).sub.4 (14.24 mg, 0.012 mmol) was added to a
degassed solution of 6-bromo-2-chloroquinoxaline (30 mg, 0.123
mmol), NaHCO.sub.3 (31.1 mg, 0.370 mmol) and (1R,3S,5R)-tert-butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (131 mg, 0.308 mmol) in
dioxane (1.0 mL) and H.sub.2O (0.2 mL) and the mixture was stirred
at 100.degree. C. for 2 h, then at 110.degree. C. for 2 h. The
reaction was diluted with MeOH, filtered and the filtrate was
purified by prep HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield
a TFA salt of tert-butyl
(1R,3S,5R)-3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-6-yl)-6-quinoxalinyl)-1H-benzimidazol-2-yl)--
2-azabicyclo[3.1.0]hexane-2-carboxylate (67.6 mg) as a yellow
solid. LC-MS retention time 2.13 min; m/z 725 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna 3.0.times.50 mm S10. Solvent A=90% water:10%
methanol: 0.1% TFA. Solvent B=10% water:90% methanol: 0.1% TFA.
Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient Time=3
min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.60
(1H, s), 8.71 (1H, s), 8.56 (1H, dd, J=8.8, 1.5 Hz), 8.45 (1H, d,
J=1.5 Hz), 8.26-8.35 (2H, m), 8.21 (1H, s), 8.06-8.11 (1H, m),
7.91-7.99 (2H, m), 5.02-5.13 (2H, m), 3.70 (2H, br s), 2.79 (2H,
dd, J=13.4, 9.2 Hz), 2.43-2.57 (2H, m), 1.83-1.96 (2H, m),
1.07-1.65 (18H, m), 0.95 (2H, m), 0.77-0.85 (2H, m).
##STR00404##
Intermediate 73
2,6-Bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-
-yl)quinoxaline
[0726] TFA (0.25 mL, 3.24 mmol) was added to a solution of a TFA
salt of tert-butyl
(1R,3S,5R)-3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-6-yl)-6-quinoxalinyl)-1H-benzimidazol-2-yl)--
2-azabicyclo[3.1.0]hexane-2-carboxylate (65 mg) in DCM (0.5 mL) and
the mixture was stirred at rt for 16 h. The volatiles were removed
and the residue was triturated with Et.sub.2O. The resulting solid
was collected via filtration funnel and rinsed with Et.sub.2O to
yield a TFA salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)quinoxaline (60 mg) as yellow solid. LC-MS retention time 1.51
min; m/z 525 [M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50
mm S10. Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10%
water:90% methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0.
Final % B=100. Gradient Time=3 min. Wavelength=220). .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 9.52 (1H, s), 8.58 (1H, s), 8.36 (1H,
s), 8.22-8.33 (2H, m), 8.30 (1H, d, J=9.5 Hz), 8.08 (1H, s),
7.76-7.94 (2H, m), 7.83 (1H, d, J=9.5 Hz), 4.80-5.05 (2H, m),
3.45-3.61 (2H, m), 2.70-2.92 (2H, m), 2.50-2.67 (2H, m), 2.00-2.18
(2H, m), 1.26 (2H, br s), 1.04 (2H, br s).
##STR00405##
##STR00406##
Intermediate 74
(S)-tert-Butyl
2-(5-(4-bromophenyl)-4-chloro-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0727] NCS (0.51 g, 3.82 mmol) was added to a solution of
tert-butyl
2-(5-(4-bromophenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(1.0 g, 2.55 mmol) in DMF (25 mL) and the mixture was heated at
50.degree. C. overnight. The volatiles were removed under vacuum.
The residue was purified by flash column chromatography
(BIOTAGE.RTM.), eluting with a gradient of 0 to 10% EtOAc/DCM to
afford the partially pure (S)-tert-butyl
2-(5-(4-bromophenyl)-4-chloro-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(1.04 g) as yellow foam. LC-MS retention time 1.99 min; m/z 427.12
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.63 (s,
4H), 4.74-4.81 (m, 1H), 3.64-3.72 (m, 1H), 3.45-3.55 (m, 1H),
2.27-2.42 (m, 1H), 2.00-2.15 (m, 2H), 1.90-2.00 (m, 1H), 1.48 (m,
3H), 1.27 (m, 6H).
##STR00407##
Intermediate 75
(S)-tert-Butyl
2-(4-chloro-5-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthal-
en-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
[0728] A solution of (S)-tert-butyl
2-(5-(4-bromophenyl)-4-chloro-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
(1.04 g, 2.44 mmol) and
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(0.926 g, 2.44 mmol) in DME (10 mL) and water (2 mL) was degassed
under vacuum for 10 min. The mixture was heated at ca
.about.80.degree. C., Pd(Ph.sub.3P).sub.4 (0.282 g, 0.244 mmol) was
added under a stream of nitrogen and the reactor was sealed. The
heating was pursued further to 130.degree. C. for 8 h. Additional
Pd(PPh.sub.3).sub.4 (100 mg) was added and the heating was pursued
overnight. The DME was removed in vacuo and the crude material was
partitioned between EtOAc/H.sub.2O. The layers were separated and
the aqueous layer was extracted several times with EtOAc. The
combined organic extracts were dried over Na.sub.2SO.sub.4 and
evaporated in vacuo. The residue was purified by flash column
chromatography (BIOTAGE.RTM.), eluting with a gradient of 0 to 100%
EtOAc/hexanes to afford the partially pure (S)-tert-butyl
2-(4-chloro-5-(4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthal-
en-2-yl)phenyl)-1H-imidazol-2-yl)pyrrolidine-1-carboxylate
contaminated with triphenylphosphine (292 mg). A small aliquot was
purified further by a reverse phase HPLC (water/acetonitrile/TFA)
to afford the pure title material. The remaining material was used
in subsequent step without further purification. LC-MS retention
time 2.227 min; m/z 598.46 (MH-) LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/10 mM ammonium acetate and Solvent B was 10%
H.sub.2O/90% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (500 MHz, MeOD, TFA salt, partial NMR) .delta. ppm 3.71
(br s, 2H), 3.55 (br s, 2H), 2.45 (br s, 1H), 2.07-2.17 (m, 2H),
1.97-2.06 (m, 1H), 1.50 (s, 3H), 1.43 (m, 6H), 1.31 (br s, 6H).
##STR00408##
Intermediate 76
(S)-tert-Butyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5--
yl)naphthalen-2-yl)phenyl)-4-chloro-1H-imidazol-2-yl)pyrrolidine-1-carboxy-
late
[0729] A solution of (S)-tert-butyl
2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (163 mg, 0.450
mmol), (S)-tert-butyl
2-(5-iodo-1H-imidazol-2-yl)pyrrolidine-1-carboxylate (163 mg, 0.450
mmol), sodium bicarbonate (113 mg, 1.350 mmol) in a solvent mixture
of DME (10 mL) and water (2 mL) was degassed under vacuum for 10
min. The mixture was heated at 80.degree. C. then
Pd(Ph.sub.3P).sub.4 (52.0 mg, 0.045 mmol) was added and the reactor
was flushed with nitrogen and sealed. The heating was pursued
further to 120.degree. C. for 16 h. The flask was cooled to room
temperature, the DME was removed in vacuo and the crude material
was partitioned between DCM/H.sub.2O. The layers were separated and
the aqueous layer was extracted several times with DCM. The
combined organic extracts were dried over Na.sub.2SO.sub.4 and
evaporated in vacuo. The residue was purified by flash column
chromatography (BIOTAGE.RTM.), eluting with a gradient of 0 to 100%
EtOAc/hexanes to afford the partially pure (S)-tert-butyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5--
yl)naphthalen-2-yl)phenyl)-4-chloro-1H-imidazol-2-yl)pyrrolidine-1-carboxy-
late contaminated with triphenylphosphine (77 mg). LC-MS retention
time 1.830 min; m/z 709.43 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.30 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and Solvent B
was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(400 MHz, chloroform-d Partial NMR) .delta. ppm 5.00 (d, J=4.5 Hz,
2H), 3.38-3.52 (m, 4H), 3.10 (br. s, 1H), 2.91 (br s, 1H),
2.09-2.33 (m, 4H), 1.93-2.05 (m, 2H), 1.54 (s, 18H).
##STR00409##
Intermediate 77
4-Chloro-2-((S)-pyrrolidin-2-yl)-5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-imida-
zol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazole
[0730] TFA (2 mL) was added to a solution of (S)-tert-butyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5--
yl)naphthalen-2-yl)phenyl)-4-chloro-1H-imidazol-2-yl)pyrrolidine-1-carboxy-
late (75 mg) in DCM (10 mL) and the mixture was stirred for 2 h at
room temperature. The solvents were removed in vacuo and the
residue was taken up in a solvent mixture of 1:1
methanol/CH.sub.2Cl.sub.2 and filtered through an MCX cartridge
(Strata XC). The cartridge was washed with methanol and the
compound was eluted with a solution of NH.sub.3 in methanol (2M).
The appropriate fractions were concentrated in vacuo to afford
4-chloro-2-((S)-pyrrolidin-2-yl)-5-(4-(6-(2-((S)-pyrrolidin-2-yl)--
1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazole (40 mg, 0.079
mmol) as yellow solid. LC-MS retention time 1.353 min; m/z 509.32
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD, partially soluble
only) .delta. ppm 8.20 (s, 1H), 8.09 (s, 1H), 7.88-7.96 (m, 2H),
7.76-7.88 (m, 6H), 7.51 (s, 1H), 4.50 (t, J=7.28 Hz, 1H), 4.35 (t,
J=7.15 Hz, 1H), 3.03-3.31 (partially masked by methanol, m, 4H),
2.24-2.42 (m, 2H), 1.88-2.21 (m, 6H).
##STR00410##
##STR00411##
Intermediate 79
(S)-tert-Butyl
2-(7-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-
naphthalen-2-yl)-4,5-dihydro-1H-naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1--
carboxylate
[0731] A solution of (S)-tert-butyl
2-(5-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
imidazol-2-yl)pyrrolidine-1-carboxylate (402 mg, 0.821 mmol),
(S)-tert-butyl
2-(7-bromo-4,5-dihydro-1H-naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carbo-
xylate (344 mg, 0.821 mmol) and sodium bicarbonate (207 mg, 2.46
mmol) in DME (14 mL) and water (1.8 mL) was degassed under vacuum
for 10 min. The mixture was heated at 80.degree. C. at which time
the flask was opened and Pd(Ph.sub.3P).sub.4 (76 mg, 0.066 mmol)
was added. The flask was flushed with nitrogen, sealed and heated
at 120.degree. C. for 16 h. The flask was cooled to room temp., the
DME was removed in vacuo and the crude material was partitioned
between DCM and H.sub.2O. The layers were separated and the aqueous
layer was extracted several times with DCM. The combined organic
extracts were dried over Na.sub.2SO.sub.4 and evaporated in vacuo.
The residue was purified by flash column chromatography
(BIOTAGE.RTM.), eluting with a gradient of 20 to 100% EtOAc/Hexanes
and then 0 to 10% methanol/DCM to afford (S)-tert-butyl
2-(7-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-
naphthalen-2-yl)-4,5-dihydro-1H-naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1--
carboxylate (183 mg). LC-MS retention time 1.543 min; m/z 701.47
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00412##
Intermediate 80
(S)-tert-Butyl
2-(7-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-
naphthalen-2-yl)-1H-naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate
[0732] Manganese(IV) oxide (1.53 g, 17.6 mmol) was added to a
solution of (S)-tert-butyl
2-(7-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-
naphthalen-2-yl)-4,5-dihydro-1H-naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1--
carboxylate (183 mg, 0.196 mmol) in DCM (5 mL) and the mixture was
stirred overnight at room temperature. The reaction mixture was
filtered through a pad of diatomaceous earth (CELITE.RTM.) and
washed with a solution of methanol/DCM 1:1. The volatiles were
removed under vacuum using a rotavap to afford (S)-tert-butyl
2-(7-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-
naphthalen-2-yl)-1H-naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate
(163 mg) as tan solid. LC-MS retention time 1.523 min; m/z 699.53
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00413##
Intermediate 81
2-((S)-Pyrrolidin-2-yl)-7-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)nap-
hthalen-2-yl)-1H-naphtho[1,2-d]imidazole
[0733] TFA (2 mL, 26.0 mmol) was added in one portion to a stirred
solution of (S)-tert-butyl
2-(7-(6-(2-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)-1H-imidazol-5-yl)-
naphthalen-2-yl)-1H-naphtho[1,2-d]imidazol-2-yl)pyrrolidine-1-carboxylate
(137 mg) in CH.sub.2Cl.sub.2 (10 mL) at room temperature. The
mixture was stirred for 2 h at room temp. and then the solvents
were removed in vacuo. The residue was taken up in 50%
methanol/CH.sub.2Cl.sub.2 and filtered through an MCX cartridge
(Strata XC). The cartridge was washed with methanol and the
compound was eluted with a solution of NH.sub.3 in methanol (2M).
The appropriate fractions were concentrated in vacuo to afford
2-((S)-pyrrolidin-2-yl)-7-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol--
5-yl)naphthalen-2-yl)-1H-naphtho[1,2-d]imidazole (98 mg, 0.20 mmol)
as orange solid. LC-MS retention time 1.245 min; m/z 499.30 (MH+).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Waters Sunfire 5u C18 4.6.times.30 mm column using
a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time
of 3 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and
Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.59 (d,
J=8.5 Hz, 1H), 8.43 (d, J=1.5 Hz, 1H), 8.36 (s, 1H), 8.31 (s, 1H),
8.13 (dd, J=8.6, 1.8 Hz, 1H), 8.01-8.10 (m, 3H), 7.90-7.98 (m, 2H),
7.87 (s, 1H), 7.80 (d, J=8.6 Hz, 1H), 5.18 (t, J=7.9 Hz, 1H), 5.05
(t, J=8.1 Hz, 1H), 3.63-3.72 (m, 1H), 3.52-3.63 (m, 3H), 2.61-2.77
(m, 2H), 2.45-2.58 (m, 2H), 2.33-2.43 (m, 2H), 2.20-2.33 (m,
2H).
##STR00414##
##STR00415##
Intermediate 82
(2S,2'S)-tert-Butyl
2,2'-(6,6'-(quinoline-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyrro-
lidine-1-carboxylate
[0734] Pd(Ph.sub.3P).sub.4 (19.06 mg, 0.016 mmol) was added to a
stirred and degassed solution of 6-bromo-2-chloroquinoline (40 mg,
0.165 mmol), (S)-tert-butyl
2-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)pyrrolidine-1-carboxylate (150 mg, 0.363 mmol) and NaHCO.sub.3
(41.6 mg, 0.495 mmol) in dioxane (1 mL) and H.sub.2O (0.2 mL). The
reaction mixture was then heated at 110.degree. C. for 2 h., cooled
to rt, diluted with MeOH and purified by prep HPLC (H.sub.2O-MeOH
with 0.1% TFA buffer) to yield a TFA salt of (2S,2'S)-tert-butyl
2,2'-(6,6'-(quinoline-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyrro-
lidine-1-carboxylate (73 mg) as a yellow solid. LC-MS retention
time 2.02 min; m/z 700 [M+H].sup.+. (Column PHENOMENEX.RTM. Luna
3.0.times.50 mm S10. Solvent A=90% water:10% methanol: 0.1% TFA.
Solvent B=10% water:90% methanol: 0.1% TFA. Flow Rate=4 mL/min.
Start % B=0. Final % B=100. Gradient Time=3 min. Wavelength=220).
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.63-8.74 (1H, m), 8.60
(1H, s), 8.44 (1H, dd, J=8.7, 1.6 Hz), 8.38 (1H, br s), 8.32 (1H,
d, J=8.8 Hz), 8.25 (2H, d, J=8.8 Hz), 8.14-8.21 (1H, m), 8.04-8.12
(1H, m), 7.90-8.00 (2H, m), 5.24-5.36 (2H, m), 3.73-3.83 (2H, m),
3.59-3.72 (2H, m), 2.57-2.74 (2H, m), 2.20-2.33 (2H, m), 2.09-2.20
(4H, m), 1.50 (9H, br s), 1.23 (9H, br s).
##STR00416##
Intermediate 83
2,6-Bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)quinoline
[0735] TFA (0.25 mL, 3.24 mmol) was added to a solution of a TFA
salt of (2S,2'S)-tert-butyl
2,2'-(6,6'-(quinoline-2,6-diyl)bis(1H-benzo[d]imidazole-6,2-diyl))dipyrro-
lidine-1-carboxylate (65 mg) in DCM (0.5 mL) and the mixture was
stirred at rt for 2 h. The volatiles were removed under vacuum and
the residue was triturated with Et.sub.2O. The resulting solid was
collected via filtration and rinsed with Et.sub.2O to yield a TFA
salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)quinoline
(61 mg) as yellow solid. LC-MS retention time 1.12 min; m/z 500
[M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.89 (1H, d, J=8.8 Hz), 8.48 (1H, d, J=1.3 Hz), 8.45
(1H, s), 8.34-8.41 (2H, m), 8.31 (1H, d, J=8.8 Hz), 8.12 (1H, dd,
J=8.5, 1.8 Hz), 8.09 (1H, s), 7.89 (1H, d, J=8.5 Hz), 7.77-7.85
(2H, m), 5.05-5.19 (2H, m), 3.44-3.72 (4H, m), 2.61-2.75 (2H, m),
2.19-2.49 (6H, m).
##STR00417##
##STR00418##
Intermediate 84
(1R,3S,5R)-tert-Butyl
3-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0736] Pd(Ph.sub.3P).sub.4 (0.382 g, 0.330 mmol) was added to a
degassed solution of
2,6-bis(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene
(2.51 g, 6.61 mmol), (1R,3S,5R)-tert-butyl
3-(5-bromo-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxyl-
ate (2.5 g, 6.6 mmol) and sodium carbonate (2.78 g, 33.0 mmol) in
DME (52.9 mL) and water (13.2 mL) at 80.degree. C. The reaction was
backfilled with nitrogen, sealed and heated at 100.degree. C.
overnight. The volatiles were removed under vacuum and the residue
was taken in EtOAc, washed with water, and the water layer was
extracted with EtOAc. The combined organic layers were filtered
through a pad of diatomaceous earth (CELITE.RTM.) and dried over
Na.sub.2SO.sub.4, concentrated and purified on a BIOTAGE.RTM. (dry
loaded on silica gel, charged to a 90 g silica gel cartridge and
eluted with a gradient of 0 to 70% ethyl acetate in hexanes, then
flushed with 10% MeOH/DCM) to yield (1R,3S,5R)-tert-butyl
3-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (1.8
g, 3.3 mmol, 49% yield) as yellowish solid. LC-MS retention time
1.905 min; m/z 552.47 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00419##
Intermediate 85
tert-Butyl
(1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl-
)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0737] A 200 mL pressure round bottom flask equipped with a
magnetic stir bar was charged with (1R,3S,5R)-tert-butyl
3-(5-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (795
mg, 1.44 mmol), (1R,3S,5R)-tert-butyl
3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(538 mg, 1.43 mmol) and sodium bicarbonate (502 mg, 5.97 mmol) in
DME (9.5 mL) and water (2.4 mL). The solution was degassed under
vacuum for 5 min and the reactor was back filled with N.sub.2.
Then, Pd(Ph.sub.3P).sub.4 (110 mg, 0.096 mmol) was added under a
stream of N.sub.2 and the reactor was sealed and the heated
overnight at 120.degree. C. The reaction was cooled to room
temperature and the volatiles were removed under vacuum. The
residue was partitioned between EtOAc and water and the aqueous
layer was extracted with EtOAc. The combined organics were filtered
through a pad of diatomaceous earth (CELITE.RTM.), dried over
Na.sub.2SO.sub.4 and the volatiles were removed under vacuum. The
residue was purified by flash column chromatography (BIOTAGE.RTM.),
eluting with a gradient of 50 to 100% EtOAc/Hexanes and then 10%
methanol/DCM to afford the partially pure target product
contaminated with starting material, reduced starting material and
other reaction side products. The residue was further purified by
prep HPLC (ACN/water, 0.1% TFA) to yield a TFA salt of tert-butyl
(1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabic-
yclo[3.1.0]hexane-2-carboxylate (200.5 mg). A side product was also
collected form the same reaction mixture which was identified as a
TFA salt
(1R,3S,5R)-3-(5-(6'-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicy-
clo[3.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-2,2'-binaphthalen-6-yl)-1H-benzi-
midazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (108 mg)
LC-MS retention time 1.497 min; m/z 673.52 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Waters Sunfire 5u C18 4.6.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and Solvent B
was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.33 (s, 1H), 8.31 (s, 1H),
8.09-8.20 (m, 3H), 7.96-8.08 (m, 2H), 7.87-7.94 (m, 2H), 5.07 (t,
J=8.2 Hz, 1H), 4.91 (m, 1H), 3.69 (br s, 2H), 2.70-2.84 (m, 2H),
2.48 (br s, 2H), 1.81-1.94 (m, 2H), 1.51 (br s, 1H), 1.51 (br s,
9H), 1.31 (br s, 5H), 1.21 (br s, 4H), 0.89-0.99 (m, 2H), 0.82 (br
s, 1H), 0.77 (br s, 1H).
##STR00420##
Intermediate 86
(1R,3S,5R)-3-(5-(6'-(2-((1R,3S,5R)-2-(tert-Butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-6-yl)-2,2'-binaphthalen-6-yl)-1H-benzimidazo-
l-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0738] Side product obtained as TFA salt (108 mg) after
purification of the reaction mixture from Intermediate 85
synthesis. LC-MS retention time 1.882 min; m/z 849.84 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Waters Sunfire 5u C18 4.6.times.30 mm column using
a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time
of 3 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and
Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.36 (s,
2H), 8.28 (s, 2H), 8.11-8.19 (m, 6H), 8.03-8.09 (m, 4H), 7.87-7.96
(m, 4H), 5.07 (t, J=8.2 Hz, 2H), 3.71 (br s, 2H), 2.75-2.85 (m,
2H), 2.51 (br s, 2H), 1.86-1.94 (m, 2H), 1.52 (m, 10H), 1.22 (br s,
8H), 0.97 (ddd, J=8.4, 6.0, 5.8 Hz, 2H), 0.83 (br s, 2H).
##STR00421##
Intermediate 87
6-(6-(2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)naphth-
alen-2-yl)-2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazo-
le
[0739] 4M HCl in dioxane (10 mL, 40.0 mmol) was added to a stirred
solution of a TFA salt of (1R,3S,5R)-tert-butyl
3-(5-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-
-yl)-1H-benzo[d]imidazol-5-yl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexane-2-carboxylate (151 mg) in DCM (3 mL). Precipitate
formed immediately. The suspension was agitated for 30 min. The
volatile components were evaporated under vacuum. The crude residue
was taken in DCM (3 mL) and TFA (2 mL) was added. The solution was
agitated for 1 h. The volatile components were evaporated under
reduced pressure. The residue was taken into MeOH and filtered
through a Strata XC MCX cartridge. The cartridge was washed with
methanol and the compound was release from the cartridge by eluting
with a solution of 2M of ammonia/methanol. The ammonia/methanol
filtrate was evaporated under reduced pressure to yield
6-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)napht-
halen-2-yl)-2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidaz-
ole (79 mg, 0.17 mmol) as yellow solid. LC-MS retention time 1.130
min; m/z 473.35 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.19 (s, 1H), 8.07 (s, 1H), 7.92 (dd, J=8.6, 5.8 Hz,
2H), 7.88 (s, 1H), 7.82 (dd, J=5.8, 1.5 Hz, 2H), 7.61-7.66 (m, 2H),
7.51 (s, 1H), 4.24-4.35 (m, 2H), 3.11 (td, J=6.1, 2.4 Hz, 1H), 3.06
(td, J=6.0, 2.6 Hz, 1H), 2.40-2.53 (m, 2H), 2.18-2.33 (m, 2H),
1.69-1.76 (m, 1H), 1.62-1.69 (m, 1H), 0.88 (ddd, J=6.6, 4.3, 2.6
Hz, 1H), 0.79-0.84 (m, 1H), 0.61-0.68 (m, 1H), 0.55-0.60 (m,
1H).
##STR00422##
Intermediate 88
6,6'-Bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
5-yl)-2,2'-binaphthyl
[0740] TFA (2.0 mL, 26 mmol) was added to a solution of a TFA salt
(1R,3S,5R)-3-(5-(6'-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3-
.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-2,2'-binaphthalen-6-yl)-1H-benzimidaz-
ol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (108 mg) in DCM (5
mL) and the reaction mixture was stirred for 2 h at rt. The
reaction mixture was then concentrated to yield a TFA salt of
6,6'-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-
-5-yl)-2,2'-binaphthyl (22 mg) as tan solid. LC-MS retention time
1.722 min; m/z 649.54 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.33 (br s, 2H), 8.22 (br s, 2H), 8.09-8.15 (m, 4H),
8.01-8.07 (m, 4H), 7.89-7.95 (m, 2H), 7.77-7.87 (m, 4H), 4.85-4.98
(m, 2H), 3.51-3.57 (m, 2H), 2.79-2.87 (m, 2H), 2.59-2.68 (m, 2H),
2.07-2.14 (m, 2H), 1.23-1.30 (m, 2H), 1.01-1.09 (m, 2H).
##STR00423##
##STR00424##
Intermediate 89
tert-Butyl
(1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-1,5-naphthyridin-2-yl)-1H-be-
nzimidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0741] Tetrakis(triphenylphosphine)palladium(0) (17.4 mg, 0.015
mmol) was added to a solution of 2,6-dichloro-1,5-naphthyridine (30
mg, 0.151 mmol), sodium bicarbonate (38.0 mg, 0.452 mmol) and
(1R,3S,5R)-tert-butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (141 mg, 0.332 mmol) in
dioxane (1.0 mL) and H.sub.2O (0.2 mL) and the mixture was heated
and stirred at 110.degree. C. for 2 h. The reaction was diluted
with MeOH, filtered, concentrated and purified by prep HPLC
(H.sub.2O-MeOH with 10 mM NH.sub.4OAc buffer) to yield tert-butyl
(1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-6-yl)-1,5-naphthyridin-2-yl)-1H-benzimidazol-
-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (22.3 mg, 0.030
mmol) as a white solid. LC-MS retention time 2.11 min; m/z 725
[M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220).
##STR00425##
Intermediate 90
2,6-Bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-
-yl)-1,5-naphthyridine
[0742] TFA (0.25 mL, 3.2 mmol) was added to a solution of
tert-butyl
(1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-6-yl)-1,5-naphthyridin-2-yl)-1H-benzimidazol-
-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (20 mg, 0.028 mmol)
in DCM (0.5 mL) and the mixture was stirred at rt for 16 h. The
volatile were removed under vacuum and the residue was triturated
with Et.sub.2O. The resulting solid was collected via filtration
and rinsed with Et.sub.2O to yield a TFA salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)-1,5-naphthyridine (26 mg, 0.022 mmol, 78% yield) as a yellow
solid. LC-MS retention time 1.41 min; m/z 525 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna 3.0.times.50 mm S10. Solvent A=90% water:10%
methanol: 0.1% TFA. Solvent B=10% water:90% methanol: 0.1% TFA.
Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient Time=3
min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.61
(2H, d, J=8.8 Hz), 8.51 (2H, s), 8.40 (2H, d, J=9.0 Hz), 8.23 (2H,
dd, J=8.5, 1.5 Hz), 7.81 (2H, d, J=8.5 Hz), 4.74-4.97 (2H, m),
3.47-3.57 (2H, m), 2.78-2.88 (2H, m), 2.52-2.65 (2H, m), 2.04-2.15
(2H, m), 1.22-1.33 (2H, m), 0.99-1.11 (2H, m).
##STR00426##
##STR00427##
Intermediate 91
tert-Butyl
(1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-5-chloro-1H-imid-
azol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0743] NCS (20 mg, 0.15 mmol) was added portionwise to a stirring
solution of tert-butyl
(1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabic-
yclo[3.1.0]hexane-2-carboxylate (183 mg, 0.272 mmol) in DMF (5 mL)
at rt. The reaction mixture was then heated at 50.degree. C. for 3
h. The reaction was cooled to 0.degree. C., additional NCS (15 mg,
0.112 mmol) was added, and the mixture was stirred for 2 h at
50.degree. C. The reaction was diluted with water (2 mL) and
methanol (2 mL), the volatiles were removed and the crude product
was purified by prep HPLC (TFA) to yield a TFA salt of tert-butyl
(1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-5-chloro-1H-imidazol-2-yl)-
-2-azabicyclo[3.1.0]hexane-2-carboxylate (140.4 mg) as orange
solid. LC-MS retention time 1.822 min; m/z 707.54 (1:1) (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Waters Sunfire C18 4.6.times.30 mm 5 .mu.m column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 4 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.27 (d,
J=2.8 Hz, 2H), 8.03-8.18 (m, 4H), 7.87-7.98 (m, 3H), 5.04-5.10 (m,
1H), 4.63 (d, J=7.6 Hz, 1H), 3.71 (br s, 1H), 3.63 (br s, 1H),
2.74-2.87 (m, 1H), 2.59 (dd, J=13.3, 8.7 Hz, 1H), 2.51 (br s, 1H),
2.41 (ddd, J=13.6, 6.7, 6.6 Hz, 1H), 1.86-1.95 (m, 1H), 1.78 (br s,
1H), 1.12-1.63 (m, 18H), 0.97 (ddd, J=8.7, 6.0, 5.8 Hz, 1H),
0.79-0.91 (m, 2H), 0.66 (br s, 1H).
##STR00428##
Intermediate 92
2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hex-3-yl)-5-(6-(2-((1R,3S,5R)-2-azabicycl-
o[3.1.0]hex-3-yl)-4-chloro-1H-imidazol-5-yl)-2-naphthyl)-1H-benzimidazole
[0744] TFA (2 mL, 26.0 mmol) was added to a solution of a TFA salt
of tert-butyl
(1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-5-chloro-1H-imidazol-2-yl)-
-2-azabicyclo[3.1.0]hexane-2-carboxylate (131 mg, 0.140 mmol) in
DCM (10 mL) and the mixture was stirred for 3 h at rt. The
volatiles were removed under vacuum and the residue was taken into
MeOH and filtered through a Strata XC MCX cartridge. The cartridge
was washed with methanol and the compound was release from the
cartridge by eluting with a solution of 2M of ammonia/methanol. The
ammonia/methanol filtrate was evaporated under reduced pressure to
yield
2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-5-(6-(2-((1R,3S,5R)-2-azabicyc-
lo[3.1.0]hex-3-yl)-4-chloro-1H-imidazol-5-yl)-2-naphthyl)-1H-benzimidazole
(55 mg, 0.11 mmol) as tan solid. LC-MS retention time 1.375 min;
m/z 507.33 (1:1) (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire C18
4.6.times.30 mm 5 .mu.m column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
methanol/90% water/0.1% TFA and Solvent B was 10% water/90%
methanol/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.26 (s, 1H), 8.19 (s, 1H), 8.00-8.09 (m, 3H), 7.92
(ddd, J=12.1, 8.6, 1.8 Hz, 2H), 7.85 (dd, J=8.6, 1.5 Hz, 1H), 7.81
(d, J=8.6 Hz, 1H), 4.96 (dd, J=11.0, 7.6 Hz, 1H), 4.64 (app t,
J=9.3 Hz, 1H), 3.53-3.59 (m, 1H), 3.44-3.50 (m, 1H), 2.82 (dd,
J=13.0, 7.5 Hz, 1H), 2.61-2.74 (m, 3H), 2.03-2.15 (m, 2H),
1.24-1.30 (m, 1H), 1.18-1.24 (m, 1H), 0.96-1.09 (m, 2H).
##STR00429##
##STR00430##
Intermediate 93
(1R,3S,5R)-3-(2-(3-Bromophenyl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate
[0745] DIPEA (2.79 mL, 16.0 mmol) was added to a stirring slurry of
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (2.00 g, 8.80 mmol) and 2-bromo-1-(3-bromophenyl)ethanone
(2.22 g, 8.00 mmol) in acetonitrile (25 mL) (the solution became
clear and amber colored) and the reaction mixture was stirred
overnight. The reaction was concentrated and purified via
BIOTAGE.RTM. (80 g SiO.sub.2, 10-25% EtOAc/hexanes) to yield
(1R,3S,5R)-3-(2-(3-bromophenyl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (3.37 g, 7.94 mmol) as
a viscous light yellow oil. LC-MS retention time 1.853 min; m/z 423
and 425.98 (1:1) (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a waters XTERRA.RTM. MS 7u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 8.04 (d, J=1.8 Hz,
1H), 7.83 (d, J=7.8 Hz, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.38 (dd,
J=8.0, 7.8 Hz, 1H), 5.58-5.10 (m, 2H), 4.23 (br s, 1H), 3.62-3.39
(m, 1H), 2.58 (dt, J=13.3, 6.5 Hz, 1H), 2.46 (dd, J=13.3, 9.5 Hz,
1H), 1.73-1.51 (m, 1H), 1.47 (s, 9H), 0.85 (br s, 1H), 0.51 (s,
1H).
##STR00431##
Intermediate 94
(1R,3S,5R)-tert-Butyl
3-(4-(3-bromophenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate
[0746] In a 350 mL high pressure vessel ammonium acetate (5.94 g,
77 mmol) was added to a solution of
(1R,3S,5R)-3-(2-(3-bromophenyl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (3.33 g, 7.85 mmol) in
xylene (75 mL) and stirred under nitrogen for 5 minutes. The vessel
was sealed and then placed into an oil bath which had been heated
to 140.degree. C. (reaction became dark brown) and the reaction was
held at 140.degree. C. for 11 h. Additional ammonium acetate (3.0
g) was added and the reaction was stirred at 145.degree. C. for 8
hrs, cooled to rt and stirred (>90% conversion by LC-MS). The
reaction was concentrated under high vacuum to a brown oil which
was partitioned between DCM (.about.200 mL) and 1/2 sat. sodium
bicarbonate (-200 mL). The organic layer was dried (MgSO.sub.4),
filtered and concentrated. The crude orange solidified foam was
purified on a BIOTAGE.RTM. Horizon (20-50% EtOAc/hexanes, 160 g
SiO.sub.2) to yield (1R,3S,5R)-tert-butyl
3-(4-(3-bromophenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate (2.03 g, 5.02 mmol) as a yellow solidified foam. LC-MS
retention time 2.450 min; m/z 404 and 406.06 (1:1) (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 7.89 (s, 1H), 7.65 (d, J=6.5 Hz, 1H), 7.42-7.32 (m 2H), 7.26
(t, J=7.8 Hz, 1H), 4.72-4.61 (m, 1H), 3.62-3.53 (m, 1H), 2.51 (dd,
J=13.0, 7.8 Hz, 1H), 2.36-2.26 (m, 1H), 1.75-1.66 (m, 1H), 1.29 (br
s, 9H), 0.84 (dt, J=8.0, 5.7 Hz, 1H), 0.63-0.57 (m, 1H).
##STR00432##
Intermediate 95
tert-Butyl
(1R,3S,5R)-3-(4-(3-(6-(2-((1S,3R,5S)-2-(tert-butoxycarbonyl)-2--
azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazo-
l-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0747] A solution of (1R,3S,5R)-tert-butyl
3-(5-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (220 mg,
0.395 mmol) (prepared in the same manner as Intermediate 51 from
Intermediate 55), (1R,3S,5R)-tert-butyl
3-(4-(3-bromophenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate (192 mg, 0.474 mmol) and cesium carbonate (386 mg, 1.185
mmol) in THF (4 mL) and water (4 mL) in a 100 mL pressure flask was
degassed under vacuum for 5 min and then backfilled with nitrogen.
Palladium (II) acetate (8.9 mg, 0.039 mmol) was added and the
mixture was heated for 4 h at 120.degree. C. The reaction mixture
was cooled to RT, diluted with water and extracted with EtOAC and
DCM. The combined organic layers were washed with brine, dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude residue
was purified on a BIOTAGE.RTM. system (90 g silica gel cartridge,
eluted with a gradient of 0 to 100% EtOAc in hexanes) to yield
tert-butyl
(1R,3S,5R)-3-(4-(3-(6-(2-((1S,3R,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2--
azabicyclo[3.1.0]hexane-2-carboxylate (142 mg, 0.203 mmol) as a
yellow solid. LC-MS retention time 1.585 min;
[0748] m/z 699.57 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire C18
4.6.times.30 mm 5 um column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
methanol/90% water/0.1% TFA and Solvent B was 10% water/90%
methanol/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.33 (d, J=1.2 Hz, 1H), 8.31 (s, 1H), 8.20-8.24 (m,
1H), 8.14-8.20 (m, 1H), 8.10-8.14 (m, 1H), 7.97-8.07 (m, 3H), 7.94
(d, J=7.9 Hz, 1H), 7.87-7.92 (m, 1H), 7.77-7.82 (m, 1H), 7.67-7.74
(m, 1H), 4.84-4.96 (m, 2H), 3.67 (br s, 2H), 2.69-2.80 (m, 2H),
2.47 (br s, 2H), 1.82-1.91 (m, 2H), 1.19-1.63 (m, 18H), 0.89-0.97
(m, 2H), 0.77 (br s, 2H).
##STR00433##
Intermediate 96
(1R,3S,5R)-3-(4-(3-(6-(2-((1S,3R,5S)-2-Azabicyclo[3.1.0]hex-3-yl)-1H-imida-
zol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
[0749] TFA (2 mL, 26.0 mmol) was added to a solution of tert-butyl
(1R,3S,5R)-3-(4-(3-(6-(2-((1S,3R,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2--
azabicyclo[3.1.0]hexane-2-carboxylate (130 mg, 0.20 mmol) in DCM (2
mL) and the mixture was stirred for 2 h at RT. The volatiles were
removed under vacuum and the crude product was purified by prep
HPLC (methanol/water, 0.1% TFA) to yield a TFA salt of
(1R,3S,5R)-3-(4-(3-(6-(2-((1S,3R,5S)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imid-
azol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(103 mg) as a tan solid. LC-MS retention time 1.303 min; m/z 499.13
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire C18 4.6.times.30 mm 5
.mu.M column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 4
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 2 min, a hold time of 1 min,
and an analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.33 (s,
1H), 8.22 (d, J=12.5 Hz, 2H), 8.02 (dd, J=8.6, 5.8 Hz, 2H),
7.90-7.97 (m, 2H), 7.75-7.82 (m, 4H), 7.58 (t, J=7.8 Hz, 1H),
4.74-4.78 (m, 2H), 3.46-3.53 (m, 2H), 2.64-2.74 (m, 4H), 2.04-2.12
(m, 2H), 1.19-1.26 (m, 2H), 1.02 (ddd, J=14.2, 7.3, 7.2 Hz,
2H).
##STR00434##
##STR00435##
Intermediate 97
(1R,3S,5R)-tert-Butyl
3-(6-(6-bromoquinoxalin-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate
[0750] Pd(Ph.sub.3P).sub.4 (28.5 mg, 0.025 mmol) was added to a
degassed solution of 6-bromo-2-chloroquinoxaline (60 mg, 0.246
mmol), (1R,3S,5R)-tert-butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (115 mg, 0.271 mmol)
and sodium bicarbonate (62.1 mg, 0.739 mmol) in dioxane (1 mL) and
H.sub.2O (0.2 mL) and the mixture was stirred at 110.degree. C. for
2 h and then at 120.degree. C. for 2 h. The reaction was diluted
with MeOH, filtered and purified by prep HPLC (H.sub.2O-MeOH with
10 mM NH.sub.4OAc buffer) to yield (1R,3S,5R)-tert-butyl
3-(6-(6-bromoquinoxalin-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate (102.2 mg, 0.202 mmol, 82% yield) as bright
yellow solid. LC-MS retention time 2.31 min; m/z 506 [M+H].sup.+.
(Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10. Solvent A=90%
water:10% methanol: 0.1% TFA. Solvent B=10% water:90% methanol:
0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient
Time=3 min. Wavelength=220).
##STR00436##
Intermediate 98
2-(2-((1R,3S,5R)-2-(tert-Butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-
-benzo[d]imidazol-6-yl)quinoxalin-6-ylboronic acid
[0751] Pd(Ph.sub.3P).sub.4 (23.28 mg, 0.020 mmol) was added to a
degassed solution of (1R,3S,5R)-tert-butyl
3-(6-(6-bromoquinoxalin-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate (102 mg, 0.201 mmol), KOAc (49.4 mg, 0.504
mmol) and bis(pinacolato)diboron (113 mg, 0.443 mmol) in dioxane (2
mL) and the reaction was stirred at 83.degree. C. for 16 h. The
reaction mixture was partitioned between EtOAc (20 mL) and sat.
NH.sub.4Cl (aq.) (5 mL). The organic layer was dried (MgSO.sub.4),
filtered and concentrated to an orange oil, which was purified by
prep HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield
2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-yl)-1-
H-benzo[d]imidazol-6-yl)quinoxalin-6-ylboronic acid (57 mg, 0.121
mmol, 60.0% yield) as orange solid. LC-MS retention time 1.94 min;
m/z 472 [M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm
S10. Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10%
water:90% methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0.
Final % B=100. Gradient Time=3 min. Wavelength=220).
##STR00437##
Intermediate 99
tert-Butyl
(1R,3S,5R)-3-(4-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-6-quinoxalinyl)-1H-imidazol--
2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0752] Pd(OAc).sub.2 (2.67 mg, 0.012 mmol) was added to a degassed
solution of
2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-yl)-1-
H-benzo[d]imidazol-6-yl)quinoxalin-6-ylboronic acid (56 mg, 0.119
mmol), (1R,3S,5R)-tert-butyl
3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(66.9 mg, 0.178 mmol),
dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine (9.76 mg, 0.024
mmol) and K.sub.2CO.sub.3 (49.3 mg, 0.356 mmol) in THF (1 mL) and
water (0.25 mL) and the mixture was stirred at 120.degree. C. for 2
h. The reaction was diluted with MeOH, filtered and purified by
prep HPLC (H.sub.2O-MeOH with 10 mM NH.sub.4OAc buffer) to yield
(1R,3S,5R)-tert-butyl
3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-
-yl)-1H-benzo[d]imidazol-6-yl)quinoxalin-6-yl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexane-2-carboxylate (32 mg, 0.047 mmol) as yellow solid.
LC-MS retention time 1.89 min; m/z 675 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna 3.0.times.50 mm S10. Solvent A=95% water/5%
methanol/10 mM ammonium acetate. Solvent B=5% water/95% methanol/10
mM ammonium acetate. Flow Rate=4 mL/min. Start % B=0. Final %
B=100. Gradient Time=2 min. Wavelength=220). .sup.1H NMR (400 MHz,
MeOD) .delta. ppm 9.44 (s, 1H), 8.45 (br s, 1H), 8.39 (s, 1H), 8.23
(d, J=8.8 Hz, 1H), 8.18 (d, J=8.5 Hz, 1H), 8.13 (d, J=8.8 Hz, 1H),
7.69-7.76 (m, 1H), 7.64 (s, 1H), 4.69-4.78 (m, 1H), 4.49-4.57 (m,
1H), 3.56-3.70 (m, 2H), 2.50-2.73 (m, 2H), 2.31-2.47 (m, 2H),
1.67-1.86 (m, 2H), 1.31 (br s, 18H), 0.82-0.95 (m, 2H), 0.54-0.75
(m, 2H).
##STR00438##
Intermediate 100
2-(2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-6-(2-(-
(1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)quinoxaline
[0753] TFA (0.25 mL, 3.24 mmol) was added to a solution of
(1R,3S,5R)-tert-butyl
3-(5-(2-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-
-yl)-1H-benzo[d]imidazol-6-yl)quinoxalin-6-yl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexane-2-carboxylate (30 mg, 0.044 mmol) in DCM (0.5 mL)
and the mixture was stirred at rt for 16 h. The volatiles were
removed under vacuum and the residue was triturated with Et.sub.2O
to yield a TFA salt of
2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6--
yl)-6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)quino-
xaline (32.7 mg) as yellow solid. LC-MS retention time 1.33 min;
m/z 475 [M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm
S10. Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10%
water:90% methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0.
Final % B=100. Gradient Time=3 min. Wavelength=220). .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 9.47 (s, 1H), 8.54 (s, 1H), 8.50 (s,
1H), 8.31 (dd, J=8.8, 1.8 Hz, 1H), 8.26 (dd, J=8.5, 1.3 Hz, 1H),
8.15 (d, J=8.8 Hz, 1H), 7.83 (s, 1H), 7.82 (d, J=8.8 Hz, 1H),
4.85-4.97 (m, 1H), 4.67 (dd, J=10.8, 7.5 Hz, 1H), 3.45-3.57 (m,
2H), 2.77-2.86 (m, 1H), 2.65-2.74 (m, 1H), 2.52-2.65 (m, 2H),
2.02-2.15 (m, 2H), 1.18-1.30 (m, 2H), 0.95-1.09 (m, 2H).
##STR00439##
##STR00440##
Intermediate 101
(1R,3S,5R)-3-(4-Iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
[0754] TFA (1 mL, 12.98 mmol) was added dropwise to a solution of
(1R,3S,5R)-tert-butyl
3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(450 mg, 1.20 mmol) in DCM (5 mL) at room temperature. The mixture
was stirred for 2 h at room temperature, then, the volatiles were
removed and the residue was taken in MeOH (5 mL) and filtered
through a Strata XC MCX cartridge. The cartridge was washed with
methanol (30 mL) and the compound was release from the cartridge by
eluting with a solution of 2M of ammonia/methanol (40 mL) and
concentrated to give
(1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(283 mg, 1.03 mmol, 86% yield) as white solid. LC-MS retention time
0.448 min; m/z 275.94 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
methanol/90% water/0.1% TFA and Solvent B was 10% water/90%
methanol/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 7.17 (s, 1H), 4.20 (dd, J=10.4, 7.3 Hz, 1H), 3.08 (td,
J=6.2, 2.6 Hz, 1H), 2.38 (dd, J=12.5, 7.3 Hz, 1H), 2.19 (ddd,
J=12.7, 10.5, 4.9 Hz, 1H), 1.68-1.74 (m, 1H), 0.85 (ddd, J=6.6,
4.4, 2.8 Hz, 1H), 0.61-0.67 (m, 1H).
##STR00441##
Intermediate 102
Methyl
(S)-1-((1R,3S,5R)-3-(4-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
xan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate
[0755] HATU (464 mg, 1.22 mmol) was added to a solution of
(1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(280 mg, 1.02 mmol), (S)-2-(methoxycarbonylamino)-3-methylbutanoic
acid (214 mg, 1.22 mmol) and DIEA (1.8 mL, 10 mmol) in DMF (3 mL).
The reaction mixture was stirred 2 h at room temperature and then
diluted with MeOH (5 mL) and water (5 mL). The volatiles were
removed under vacuum and the residue was purified with flash
chromatography (sample was dry loaded on silica gel and eluted with
0-100 ethyl acetate/hexanes) to afford methyl
(S)-1-((1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-2--
yl)-3-methyl-1-oxobutan-2-ylcarbamate (500 mg, 0.925 mmol, 91%
yield) as yellowish oil. LC-MS retention time 0.850 min; m/z 432.97
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.09 (s,
1H), 5.09 (dd, J=8.9, 4.6 Hz, 1H), 4.47-4.60 (m, 1H), 3.67 (s, 3H),
3.59-3.62 (m, 1H), 2.39-2.49 (m, 1H), 2.29-2.39 (m, 1H), 2.12 (dq,
J=13.6, 6.8 Hz, 1H), 1.95-2.06 (m, 1H), 1.11 (dt, J=8.6, 5.5 Hz,
1H), 0.94-1.02 (m, 3H), 0.91 (d, J=6.7 Hz, 3H), 0.76 (br s,
1H).
##STR00442##
Intermediate 103
(1R,3S,5R)-tert-Butyl
3-(6-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)--
2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)-1H-benzo[-
d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0756] A solution of (1R,3S,5R)-tert-butyl
3-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (573
mg, 1.04 mmol), methyl
(S)-1-((1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-2--
yl)-3-methyl-1-oxobutan-2-ylcarbamate (374 mg, 0.87 mmol),
dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine (SPHOS, 71.0
mg, 0.173 mmol) and cesium carbonate (846 mg, 2.60 mmol) in THF
(7.9 mL) and water (0.8 mL) was degassed at 0.degree. C. under
vacuum for 5 min and the reactor was then back filled with
nitrogen. Palladium(II) acetate (19.4 mg, 0.087 mmol) was added and
the reaction mixture was heated at 100.degree. C. for 3 h. water
was added to the cooled solution and the reaction was further
diluted with EtOAc (15 mL). The phases were separated and the
aqueous layer was extracted with EtOAc (15 mL). The combined
organic layers were dried over Na.sub.2SO.sub.4, filtered through a
pad of diatomaceous earth (CELITE.RTM.) and the volatiles were
removed under reduced pressure. The residue was purified with flash
chromatography (sample was dry loaded on silica gel and eluted with
20-100% ethyl acetate/hexanes then 5% MeOH/DCM) to afford
(1R,3S,5R)-tert-butyl
3-(6-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)--
2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)-1H-benzo[-
d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (246 mg,
0.337 mmol, 39.0% yield). LC-MS retention time 1.448 min; m/z
730.21 (MH+).LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.32 (br
s, 1H), 8.30 (s, 1H), 8.03-8.19 (m, 4H), 7.96-8.03 (m, 1H), 8.01
(s, 1H), 7.85-7.96 (m, 2H), 5.18 (dd, J=9.0, 7.0 Hz, 1H), 5.09 (dd,
J=8.9, 7.6 Hz, 1H), 4.58 (d, J=6.5 Hz, 1H), 3.81-3.89 (m, 1H),
3.65-3.75 (m, 1H), 3.69 (s, 3H), 2.68-2.86 (m, 2H), 2.52 (dt,
J=13.6, 6.8 Hz, 2H), 2.15-2.29 (m, 1H), 2.06-2.15 (m, 1H),
1.85-1.95 (m, 1H), 1.51 (br s, 4H), 1.41 (d, J=16.3 Hz, 1H), 1.27
(s, 1H), 1.20 (br s, 3H), 1.12 (ddd, J=8.7, 6.0, 5.9 Hz, 1H), 1.04
(d, J=6.8 Hz, 3H), 0.87-1.00 (m, 5H), 0.83 (br s, 1H).
##STR00443##
Intermediate 104
Methyl
(S)-1-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-
-yl)-1H-benzo[d]imidazol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate
[0757] TFA (2 mL,) was added to a solution of (1R,3S,5R)-tert-butyl
3-(6-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)--
2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)-1H-benzo[-
d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (226 mg,
0.310 mmol) in DCM (5 mL) and the reaction was stirred for 2 h at
room temperature. The solution was concentrated under vacuum and
the residue was taken in MeOH (10 mL) and filtered through a Strata
XC MCX cartridge and washed with methanol (25 mL). The compound was
released from the cartridge by washing the column with a solution
of 2M of ammonia/methanol (10 mL) and concentrated under reduced
pressure to give a methyl
(S)-1-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1-
H-benzo[d]imidazol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (131 mg, 0.208
mmol) as tan solid. LC-MS retention time 1.235 min; m/z 630.17
(MH+).LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.16 (s,
1H), 8.07 (s, 1H), 7.92 (dd, J=8.3, 6.3 Hz, 2H), 7.87 (s, 1H),
7.78-7.85 (m, 2H), 7.60-7.69 (m, 2H), 7.44 (s, 1H), 5.20 (dd,
J=8.8, 4.8 Hz, 1H), 4.62 (d, J=6.8 Hz, 1H), 4.28 (dd, J=9.8, 7.8
Hz, 1H), 3.66-3.76 (m, 4H), 3.35-3.41 (m, 2H), 3.03 (td, J=6.0, 2.5
Hz, 1H), 2.40-2.59 (m, 3H), 2.12-2.25 (m, 2H), 1.99-2.09 (m, 1H),
1.59-1.68 (m, 1H), 1.10-1.18 (m, 1H), 1.00-1.08 (m, 3H), 1.04 (d,
J=6.8 Hz, 3H), 0.91-1.04 (m, 1H), 0.95 (d, J=6.8 Hz, 3H), 0.76-0.84
(m, 2H), 0.50-0.59 (m, 1H).
##STR00444##
##STR00445##
Intermediate 105
Methyl
(S)-2-((1R,3S,5R)-3-(4-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
xan-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethylcarbamate
[0758] HATU (846 mg, 2.23 mmol) was added to a solution of
(1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(510 mg, 1.85 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (483 mg, 2.23 mmol) and DIEA (3.24 mL, 18.5 mmol) in DMF (5
mL) and the reaction was stirred 2 h at room temperature. The
reaction mixture was diluted with MeOH (5 mL) and water (5 mL). The
volatiles were removed under vacuum and the residue was purified
with flash chromatography (sample was dry loaded on silica gel and
eluted with 0-100% ethyl acetate/hexanes then 10% MeOH/DCM) to
afford the methyl
(S)-2-((1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-2--
yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethylcarbamate (370 mg, 0.780
mmol, 42.1% yield) as white foam. Impure material was further
purified on reverse phase HPLC (water/methanol, 0.1% TFA) to afford
the TFA salt of methyl
(S)-2-((1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
exan-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethylcarbamate, (387
mg) as colorless oil. LC-MS retention time 0.690 min; m/z 474.95
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 7.58 (s, 1H), 5.04 (dd, J=9.2, 6.7 Hz, 1H), 4.56 (d, J=7.8 Hz,
1H), 3.96 (td, J=11.7, 3.1 Hz, 2H), 3.74-3.81 (m, 1H), 3.65-3.73
(m, 3H), 3.35-3.44 (m, 2H), 2.61 (dd, J=13.6, 9.3 Hz, 1H), 2.37
(ddd, J=13.6, 6.7, 6.5 Hz, 1H), 1.96-2.09 (m, 2H), 1.37-1.62 (m,
4H), 1.06 (dt, J=8.8, 5.9 Hz, 1H), 0.84 (td, J=5.6, 2.4 Hz,
1H).
##STR00446##
Intermediate 106
(1R,3S,5R)-tert-Butyl
3-(6-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-p-
yran-4-yl)acetyl)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)naphthal-
en-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0759] A slurry of (1R,3S,5R)-tert-butyl
3-(6-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)-1H--
benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (407
mg, 0.738 mmol), a TFA salt of methyl
(S)-2-((1R,3S,5R)-3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-2--
yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethylcarbamate (350 mg),
dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine (60.6 mg, 0.148
mmol) and cesium carbonate (721 mg, 2.21 mmol) in THF (6.7 mL) and
water (0.7 mL) was degassed at 0.degree. C. under vacuum for 5 min
and then the reactor was back filled with nitrogen. Palladium(II)
acetate (16.6 mg, 0.074 mmol) was added and the mixture was heated
at 100.degree. C. for 3 h. The volatiles were removed under vacuum
and the mixture was diluted with water (20 mL) and EtOAc (20 mL).
The phases were separated and the aqueous layer was extracted with
EtOAc (20 mL). The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered and evaporated under vacuum. The residue
was purified with flash chromatography (sample was dry loaded on
silica gel and eluted with 20-100% ethyl acetate/hexanes, then 10%
MeOH/DCM). The residue was purified again by on reverse phase HPLC
(water/methanol, 0.1% TFA) to afford the TFA salt of
(1R,3S,5R)-tert-butyl
3-(6-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-p-
yran-4-yl)acetyl)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthal-
en-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(231 mg) as yellow solid. LC-MS retention time 1.395 min; m/z
772.31 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.31 (s, 2H), 8.09-8.18 (m, 3H), 8.05 (dd, J=8.7, 1.7 Hz, 1H),
7.97-8.02 (m, 2H), 7.91 (d, J=8.6 Hz, 1H), 7.88 (dd, J=8.7, 1.7 Hz,
1H), 5.16 (dd, J=9.2, 7.0 Hz, 1H), 5.01-5.09 (m, 1H), 4.62 (d,
J=7.6 Hz, 1H), 3.91-4.02 (m, 3H), 3.83-3.89 (m, 1H), 3.70 (br s,
4H), 3.35-3.45 (m, 2H), 2.68-2.83 (m, 2H), 2.45-2.57 (m, 2H),
2.04-2.16 (m, 2H), 1.90 (br s, 1H), 1.43-1.61 (m, 8H), 1.14-1.31
(m, 4H), 1.07-1.13 (m, 1H), 0.97 (dt, J=8.8, 5.8 Hz, 1H), 0.91 (br
s, 1H), 0.82 (br s, 1H).
##STR00447##
Intermediate 107
Methyl
(S)-2-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-
-yl)-1H-benzo[d]imidazol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexan-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethylcarbamate
[0760] TFA (2 mL, 26.0 mmol) was added to a solution of a TFA salt
of (1R,3S,5R)-tert-butyl
3-(6-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-p-
yran-4-yl)acetyl)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthal-
en-2-yl)-1H-benzo[d]imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(150 mg) in DCM and the mixture was stirred for 2 h at rt. The
volatiles were removed under vacuum and the residue was taken in
MeOH (15 mL), filtered through a Strata XC MCX cartridge (1 g) and
washed with methanol. The compound was release from the cartridge
by washing the column with a solution of 2M of ammonia/methanol (20
mL) and concentrated to give methyl
(S)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1-
H-benzo[d]imidazol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexan-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethylcarbamate
(100 mg, 0.149 mmol) as white solid. LC-MS retention time 1.772
min; m/z 670.36 (M-H+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire C18
4.6.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
methanol/90% water/10 mM ammonium acetate and solvent and Solvent B
was 10% water/90% methanol/10 mM ammonium acetate and solvent. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.28 (s, 1H), 8.24 (s, 1H), 8.13 (d, J=8.9 Hz, 1H), 8.05-8.10
(m, 1H), 8.03 (s, 1H), 7.96-8.00 (m, 2H), 7.84 (dd, J=8.7, 1.7 Hz,
1H), 7.75-7.82 (m, 2H), 5.16 (dd, J=9.2, 7.3 Hz, 1H), 4.84-4.88
(partially shaded by MeOD, m, 1H), 4.62 (d, J=7.6 Hz, 1H),
3.91-4.01 (m, 2H), 3.84-3.89 (m, 1H), 3.66-3.76 (m, 3H), 3.51-3.56
(m, 1H), 3.35-3.46 (m, 2H), 2.82 (dd, J=12.8, 7.3 Hz, 1H), 2.73
(dd, J=13.7, 9.2 Hz, 1H), 2.61 (td, J=12.0, 4.7 Hz, 1H), 2.53 (ddd,
J=13.8, 6.8, 6.6 Hz, 1H), 2.06-2.16 (m, 3H), 1.58-1.64 (m, 1H),
1.44-1.57 (m, 3H), 1.23-1.30 (m, 1H), 1.08-1.16 (m, 1H), 1.05 (q,
J=7.8 Hz, 1H), 0.88-0.93 (m, 1H).
##STR00448##
##STR00449##
Intermediate 108
6-Bromo-2-(1-ethoxyvinyl)quinoxaline
[0761] Dichlorobis(triphenylphosphine)-palladium(II) (17.30 mg,
0.025 mmol) was added to a solution of 6-bromo-2-chloroquinoxaline
(60 mg, 0.246 mmol) and tributyl(1-ethoxyvinyl)stannane (107 mg,
0.296 mmol) in dioxane (1.5 mL) and the mixture was stirred at
110.degree. C. for 2 h. The reaction was diluted with MeOH, and
purified by prep HPLC (H.sub.2O-MeOH with 10 mM NH.sub.4OAc buffer)
to yield 6-bromo-2-(1-ethoxyvinyl)quinoxaline (36 mg, 0.129 mmol,
52.3% yield) as white solid. LC-MS retention time 2.76 min; m/z 279
[M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220).
##STR00450##
Intermediate 109
2-Bromo-1-(6-bromoquinoxalin-2-yl)ethanone
[0762] NBS (55.1 mg, 0.310 mmol) was added to a solution of
6-bromo-2-(1-ethoxyvinyl)quinoxaline (72 mg, 0.26 mmol) in THF (2
mL) and water (0.500 mL) and the mixture was stirred at rt for 2 h.
The reaction was diluted with MeOH and purified by prep HPLC
(H.sub.2O-MeOH with 10 mM NH.sub.4OAc buffer) to yield
2-bromo-1-(6-bromoquinoxalin-2-yl)ethanone (50 mg, 0.15 mmol, 59%
yield) as white solid. LC-MS retention time 2.40 min; m/z 329
[M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220).
##STR00451##
Intermediate 110
(1S,3S,5S)-3-(2-(6-Bromoquinoxalin-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate
[0763] DIPEA (0.040 mL, 0.227 mmol) was added to a solution of
2-bromo-1-(6-bromoquinoxalin-2-yl)ethanone (50 mg, 0.152 mmol) and
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (37.9 mg, 0.167 mmol) in acetonitrile (1.5 mL) and the mixture
was stirred at rt for 16 h. The solvent was evaporated and the
residue was partitioned between EtOAc (20 mL)/aq. sat. NaHCO.sub.3
(5 mL). The organic layer was dried (MgSO.sub.4), filtered and
concentrated to yield crude
(1R,3S,5R)-3-(2-(6-bromoquinoxalin-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (63 mg) as orange
solid. LC-MS retention time 2.74 min; m/z 476 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna 3.0.times.50 mm S10. Solvent A=90% water:10%
methanol: 0.1% TFA. Solvent B=10% water:90% methanol: 0.1% TFA.
Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient Time=3
min. Wavelength=220).
##STR00452##
Intermediate 111
(1R,3S,5R)-tert-Butyl
3-(4-(6-bromoquinoxalin-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate
[0764] NH.sub.4OAc (102 mg, 1.323 mmol) was added to a solution of
(1R,3S,5R)-3-(2-(6-bromoquinoxalin-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (63 mg, 0.13 mmol) in
toluene (3 mL) and the mixture was stirred at 120.degree. C. for 4
h. Additional NH.sub.4OAc (102 mg, 1.323 mmol) was added and the
reaction was further heated at 120.degree. C. for 4 h. The reaction
was diluted with EtOAc (20 mL) and aq. sat. NaHCO.sub.3 (5 mL). The
organic layer was dried (MgSO.sub.4), filtered, concentrated and
then purified by flash silica chromatography (eluted with 1:2
EtOAc/hexane) to yield (1R,3S,5R)-tert-butyl
3-(4-(6-bromoquinoxalin-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (25 mg, 0.055 mmol) as red orange solid. LC-MS
retention time 2.07 min; m/z 456 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna 3.0.times.50 mm S10. Solvent A=90% water:10%
methanol: 0.1% TFA. Solvent B=10% water:90% methanol: 0.1% TFA.
Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient Time=3
min. Wavelength=220).
##STR00453##
Intermediate 112
(1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-(tert-Butoxycarbonyl)-2-azabicyclo[3.1-
.0]hex-3-yl)-1H-benzimidazol-5-yl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-aza-
bicyclo[3.1.0]hexane-2-carboxylate
[0765] Pd(OAc).sub.2 (1.230 mg, 5.48 .mu.mol) was added to a
degassed suspension of (1R,3S,5R)-tert-butyl
3-(4-(6-bromoquinoxalin-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (25 mg, 0.055 mmol), K.sub.2CO.sub.3 (22.71 mg,
0.164 mmol), dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine
(4.50 mg, 10.96 .mu.mol) and (1R,3S,5R)-tert-butyl
3-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[d]imidazol-2--
yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (30.3 mg, 0.071 mmol)
in THF (1 mL) and water (0.250 mL) and the mixture was stirred at
110.degree. C. for 2 h. The reaction was diluted with MeOH,
filtered and purified by prep HPLC (H.sub.2O-MeOH with 10 mM
NH.sub.4OAc buffer) to yield (1R,3S,5R)-tert-butyl
3-(5-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-
-yl)-1H-benzo[d]imidazol-6-yl)quinoxalin-2-yl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexane-2-carboxylate (12 mg, 0.018 mmol, 33% yield) as
yellow solid. LC-MS retention time 1.90 min; m/z 675 [M+H].sup.+.
(Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10. Solvent A=95%
water/5% methanol/10 mM ammonium acetate. Solvent B=5% water/95%
methanol/10 mM ammonium acetate. Flow Rate=4 mL/min. Start % B=0.
Final % B=100. Gradient Time=2 min. Wavelength=220). .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 9.38 (s, 1H), 8.28 (d, J=2.0 Hz, 1H),
8.16-8.22 (m, 1H), 8.10-8.15 (m, 1H), 7.95 (s, 2H), 7.66-7.76 (m,
2H), 3.54-3.73 (m, 2H), 2.52-2.69 (m, 2H), 2.33-2.47 (m, 2H),
1.71-1.84 (m, 2H), 1.08-1.57 (m, 20H), 0.83-0.95 (m, 2H), 0.60-0.74
(m, 2H).
##STR00454##
Intermediate 113
6-(2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-yl)--
2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)quinoxali-
ne
[0766] TFA (0.25 mL, 3.24 mmol) was added to a solution of
(1R,3S,5R)-tert-butyl
3-(5-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexan-3-
-yl)-1H-benzo[d]imidazol-6-yl)quinoxalin-2-yl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexane-2-carboxylate (10 mg, 0.015 mmol) in DCM (0.5 mL)
and the mixture was stirred at rt for 16 h. The volatiles were
removed under vacuum and the residue was triturated with Et.sub.2O.
The resulting solid was rinsed with Et.sub.2O to yield a TFA salt
of
6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-yl)-
-2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)quinoxal-
ine (10.5 mg) as yellow solid. LC-MS retention time 1.32 min; m/z
475 [M+H].sup.+. (Column PHENOMENEX.RTM. Luna 3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220).
##STR00455##
Intermediate 114
Benzyl
(2S)-2-(6-(6-(2-((2S)-1-((benzyloxy)carbonyl)-2-pyrrolidinyl)-1H-im-
idazo[4,5-b]pyridin-6-yl)-2-naphthyl)-3H-imidazo[4,5-b]pyridin-2-yl)-1-pyr-
rolidinecarboxylate
[0767] Prepared in a similar manner as Intermediate 39 using
(S)-1-(benzyloxycarbonyl)pyrrolidine-2-carboxylic acid as a
starting material rather than
(S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid and the
intermediates were not SEM protected. Prepared benzyl
(2S)-2-(6-(6-(2-((2S)-1-((benzyloxy)carbonyl)-2-pyrrolidinyl)-1H-imidazo[-
4,5-b]pyridin-6-yl)-2-naphthyl)-3H-imidazo[4,5-b]pyridin-2-yl)-1-pyrrolidi-
necarboxylate (20.8 mg) as a white solid. LC-MS retention time
1.742 min; m/z 769.41 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and Solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00456##
##STR00457##
Intermediate 114a
1-(4-(6-Acetylnaphthalen-2-yl)phenyl)ethanone
[0768] A solution of sodium carbonate (5.43 g, 51.3 mmol) in water
(35 mL) was added to a stirred solution of
1-(6-bromonaphthalen-2-yl)ethanone (2.554 g, 10.25 mmol) and
4-acetylphenylboronic acid (2.017 g, 12.30 mmol) in toluene (35.0
mL) and ethanol (35.0 mL) and nitrogen was bubbled through the
reaction mixture for 15 min. Then Pd(PPh.sub.3).sub.4 (0.237 g,
0.205 mmol) was added, and the reaction was flushed with nitrogen
and heated at 95.degree. C. under nitrogen for 8 h and then stirred
overnight at rt. The reaction was concentrated to dryness under
high vacuum and then partitioned between DCM (-200 mL) and water
(.about.150 mL). The organic layer was then washed with brine
(.about.100 mL), dried (MsSO.sub.4), filtered and concentrated. The
residue was triturated with MeOH (.about.120 mL) and the remaining
solids were redissolved into DCM and concentrated to dryness
(.about.2.7 g of orange solid). This material was dissolved into
hot EtOAc (80 mL) and allowed to cool. The resulting solids were
collected by filtration and rinsed with Et.sub.2O to yield
1-(4-(6-acetylnaphthalen-2-yl)phenyl)ethanone (1.84 g). LC-MS
retention time 2.443 min; m/z 288.98 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode. .sup.1H NMR (400 MHz, chloroform-d)
.delta. ppm 8.51 (s, 1H), 8.05-8.16 (m, 5H), 7.98 (d, J=8.8 Hz,
1H), 7.81-7.88 (m, 3H), 2.76 (s, 3H), 2.68 (s, 3H).
##STR00458##
Intermediate 114b
2-Bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone
[0769] A solution of bromine (0.669 mL, 13.0 mmol) in DCM (10 mL)
was added to a stirred solution of
1-(4-(6-acetylnaphthalen-2-yl)phenyl)ethanone (1.827 g, 6.34 mmol)
in DCM (30 mL) and the reaction was stirred at rt for 1 d. The
reaction mixture was diluted with DCM (.about.20 mL) and
concentrated to dryness to yield
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone
(2.83 g) which was used without further purification. LC-MS
retention time 2.708 min; m/z 446.71 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode. .sup.1H NMR (400 MHz, chloroform-d)
.delta. ppm 8.57 (s, 1H), 8.07-8.17 (m, 5H), 8.02 (d, J=8.8 Hz,
1H), 7.88 (d, J=8.8 Hz, 3H), 4.60 (s, 2H), 4.51 (s, 2H).
##STR00459##
Intermediate 115
(2S,5S)-2-(2-(4-(6-(2-((2S,5S)-1-(tert-Butoxycarbonyl)-5-methylpyrrolidine-
-2-carbonyloxy)acetyl)naphthalen-2-yl)phenyl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate
[0770] Hunig's Base (1.662 mL, 9.52 mmol) was added to a stirred
slurry of
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone
(1.42 g, 3.17 mmol) and
(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic
acid (1.45 g, 6.34 mmol) in acetonitrile (60 mL). The reaction was
stirred at rt for 1 d (slowly became clear) and the clear orange
solution was concentrated and purified on a BIOTAGE.RTM. Horizon
(160 g SiO.sub.2, 30-40% EtOAc/hexanes) to yield
(2S,5S)-2-(2-(4-(6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-
e-2-carbonyloxy)acetyl)naphthalen-2-yl)phenyl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate (1.81 g) as an off-white
solidified foam. LC-MS retention time 3.126 min; m/z 765.87 (M+Na).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 4 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min, and an
analysis time of 4 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 8.48 (s, 1H), 8.13
(s, 1H), 7.99-8.11 (m, 5H), 7.86 (d, J=8.3 Hz, 3H), 5.24-5.76 (m,
4H), 4.41-4.59 (m, 2H), 3.93-4.11 (m, 2H), 2.28-2.42 (m, 4H), 2.11
(br. s., 2H), 1.77 (br. s., 2H), 1.49 (br. s., 9H), 1.48 (br. s.,
9H), 1.35 (br. s., 6H).
##STR00460##
Intermediate 116
tert-Butyl
(2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methyl-
-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5--
methyl-1-pyrrolidinecarboxylate
[0771] Ammonium acetate (3.76 g, 48.7 mmol) was added to a stirred
solution of
(2S,5S)-2-(2-(4-(6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-
e-2-carbonyloxy)acetyl)naphthalen-2-yl)phenyl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate (1.81 g, 2.437 mmol) in
toluene (40 mL) and the slurry was stirred at rt for 10 min before
being heated at 100.degree. C. for 12 h. The reaction was cooled to
rt, concentrated to dryness and the residue was partitioned between
DCM (.about.150 mL) and 1/2 sat NaHCO.sub.3 (aq) (.about.150 mL).
The organic layer was washed with brine (.about.100 mL), dried
(MgSO.sub.4), filter and concentrated to a solidified brown foam
which was purified by BIOTAGE.RTM. Horizon (160 g SiO.sub.2, 1.5-3%
MeOH/DCM) to yield tert-butyl
(2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-p-
yrrolidinecarboxylate (1.46 g) as a yellow-orange solidified foam.
LC-MS retention time 2.92 min; m/z 703.32 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode.
##STR00461##
Intermediate 117
2-((2S,5S)-5-Methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
[0772] TFA (0.500 mL, 6.49 mmol) was added dropwise to a stirred
solution of tert-butyl
(2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-p-
yrrolidinecarboxylate (228 mg, 0.324 mmol) in DCE (6 mL). During
the addition, precipitate formed which hindered the stirring. DCM
(.about.10 mL) was added and the slurry was stirred while the
remaining TFA was added. The reaction was stirred at rt for 30 min,
additional TFA was added (.about.0.5 mL) and the reaction was
stirred 3 h. The reaction was concentrated to yield a TFA salt of
2-((2S,5S)-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole (102 mg,
113 mg, 114 mg) as an orange solid. LC-MS retention time 2.756 min;
m/z 503.14 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.32 (s, 1H), 8.17 (s, 1H),
8.01 (app t, J=8.2 Hz, 2H), 7.85-7.95 (m, 6H), 7.80 (s, 1H), 7.73
(s, 1H), 5.02 (app q, J=8.0 Hz, 2H), 3.92 (dddd, J=8.7, 7.0, 6.9,
4.8 Hz, 2H), 2.52-2.67 (m, 4H), 2.38-2.49 (m, 2H), 1.95-2.07 (m,
2H), 1.55 (app dd, J=6.7, 4.4 Hz, 6H).
##STR00462##
##STR00463##
Intermediate 118
2-Bromo-1-(6-bromonaphthalen-2-yl)ethanone
[0773] A solution of bromine (0.682 mL, 13.3 mmol) in DCM (20 mL)
was added to a solution of 1-(6-bromonaphthalen-2-yl)ethanone (3.30
g, 13.3 mmol) (>90% purity) in DCM (60 mL) and the reaction was
stirred at rt overnight. The reaction mixture was concentrated to
yield 2-bromo-1-(6-bromonaphthalen-2-yl)ethanone (4.35 g) as an off
white solid which was used without further purification. LC-MS
retention time 2.177 min; m/z 342.92 (MNa.sup.+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 2.0.times.30 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 1 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time
of 3 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz,
chloroform-d) .delta. ppm 8.49 (s, 1H), 8.03-8.10 (m, 2H), 7.86 (d,
J=8.5 Hz, 1H), 7.85 (d, J=8.8 Hz, 1H), 7.67 (dd, J=8.8, 2.0 Hz,
1H), 4.56 (s, 2H).
##STR00464##
Intermediate 119
(1R,3S,5R)-3-(2-(6-Bromonaphthalen-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate
[0774] DIPEA (3.47 mL, 19.88 mmol) was added to a stirred slurry of
crude 2-bromo-1-(6-bromonaphthalen-2-yl)ethanone (4.35 g, 13.3
mmol) and
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (3.01 g, 13.25 mmol) in acetonitrile (80 mL) and the reaction
was stirred at rt overnight. The reaction was concentrated to
dryness and purified by BIOTAGE.RTM. Horizon (160 g SiO.sub.2,
10-20% EtOAc/hexanes). The fractions containing the desired product
were allowed to stand for 3 d. Some fractions had crashed large
crystals (collected 240 mg, pure desired product by .sup.1H NMR).
All fractions containing the desired product were collected and
concentrated to a yellow solidified foam which was slurried with
Et.sub.2O (.about.40 mL). The white solid that formed was collected
by filtration and rinsed with Et.sub.2O to yield
(1R,3S,5R)-3-(2-(6-bromonaphthalen-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (3.24 g). LC-MS
retention time 2.760 min; m/z 472, 474.02 (1:1) (MH-). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode. .sup.1H NMR (400 mHz, MeOD) d ppm 8.61 (br
s, 1H), 8.17 (s, 1H), 8.03 (d, J=8.6 Hz, 1H), 8.00 (d, J=8.9 Hz,
1H), 7.94 (d, J=8.9 Hz, 1H), 7.71 (dd, J=8.6, 1.8 Hz, 1H),
5.75-5.66 (m, 1H), 5.61-5.46 (m, 1H), 4.34-4.26 (m, 1H), 3.46 (br
s, 1H), 2.69-2.59 (m, 1H), 2.55-2.44 (m, 1H), 1.73 (br s, 1H),
1.54-1.43 (m, 9H), 0.88 (br s, 1H), 0.59-0.53 (m, 1H).
##STR00465##
Intermediate 120
(1R,3S,5R)-tert-Butyl
3-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate
[0775]
(1R,3S,5R)-3-(2-(6-Bromonaphthalen-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (3.46 g, 7.28 mmol) and
ammonium acetate (11.2 g, 146 mmol) were dissolved into toluene
(100 mL) and stirred. Then the reaction was placed into an oil bath
which had been preheated to 100.degree. C. and stirred at that
temperature for 12 h. The reaction was allowed to cool to rt,
concentrated and partitioned between DCM (.about.200 mL) and 1/2
sat. aq. NaHCO.sub.3 (.about.150 mL). The organic layer was washed
with brine (.about.100 mL), dried (MgSO.sub.4), filtered and
concentrated to a solidified tan foam. This material was purified
on a BIOTAGE.RTM. Horizon (160 g SiO.sub.2, loaded with DCM, 30-50%
EtOAc/hexanes) to yield (1R,3S,5R)-tert-butyl
3-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (3.05 g) as a solidified yellow foam. LC-MS
retention time 2.238 min; m/z 452.07, 454.02 (1:1) (MH-). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 1 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time
of 3 min where Solvent A was 5% acetonitrile/95% H.sub.2O/10 mM
ammonium acetate and Solvent B was 5% H.sub.2O/95% acetonitrile/10
mM ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz,
chloroform-d) .delta. ppm 8.14 (br. s., 1H), 7.96 (d, J=1.8 Hz,
1H), 7.79 (d, J=9.0 Hz, 1H), 7.72 (d, J=9.8 Hz, 2H), 7.53 (dd,
J=8.8, 2.0 Hz, 1H), 7.38 (s, 1H), 4.91 (dd, J=9.0, 5.3 Hz, 1H),
3.25-3.56 (m, 2H), 2.40-2.51 (m, 1H), 1.75-1.85 (m, 1H), 1.38 (br.
s., 1H), 0.86-0.93 (m, 1H), 0.47-0.55 (m, 1H).
##STR00466##
##STR00467##
Intermediate 121
(1R,3S,5R)-tert-Butyl
3-(4-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-
e-2-carboxylate
[0776] Nitrogen was bubbled through a solution of
(1R,3S,5R)-tert-butyl
3-(5-iodo-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(1.65 g, 4.40 mmol) and Cu(I)I (168 mg, 0.880 mmol) in
triethylamine (3.07 mL, 22.0 mmol) and DMF (40 mL) for 20 min.
Then, ethynyltrimethylsilane (2.16 g, 22.0 mmol) and
Pd(PPh.sub.3).sub.4 (254 mg, 0.220 mmol) were added, the reaction
was flushed with nitrogen, sealed and stirred at rt for 24 h. The
reaction was concentrated to a brown oil, and purified by
BIOTAGE.RTM.Horizon (160 g SiO.sub.2, 20-40% EtOAc/hexanes) to
yield (1R,3S,5R)-tert-butyl
3-(5-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-
e-2-carboxylate (980 mg) as a yellow solid. LC-MS retention time
3.230 min; m/z 346.17 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, chloroform-d) d ppm 7.18 (s, 1H), 4.77
(dd, J=9.2, 5.1 Hz, 1H), 3.20 (br. s., 1H), 3.14-3.39 (m, 1H), 2.33
(dd, J=13.2, 9.4 Hz, 1H), 1.69-1.78 (m, 1H), 1.34-1.56 (m, 1H),
1.49 (s, 9H), 0.85 (dt, J=8.5, 5.7 Hz, 1H), 0.41-0.47 (m, 1H),
0.22-0.24 (m, 9H).
##STR00468##
Intermediate 122
(1R,3S,5R)-tert-Butyl
3-(4-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0777] Potassium carbonate (194 mg, 1.40 mmol) was added to a
solution of (1R,3S,5R)-tert-butyl
3-(5-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-
e-2-carboxylate (970 mg, 2.81 mmol) in MeOH (30 mL) and the
reaction was stirred under nitrogen and then heated at 50.degree.
C. (bath temp) for 4 h. The reaction was concentrated to .about.5
mL of volume, diluted with DCM (.about.40 mL) and washed with 1/2
sat brine (.about.20 mL). The organics were dried (MgSO.sub.4)
filtered and concentrated to a tan solid. This material was
purified by BIOTAGE.RTM. Horizon (40 g SiO.sub.2, loaded with DCM,
35-45% EtOAc/hexanes) to yield (1R,3S,5R)-tert-butyl
3-(5-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(747 mg) as a light yellow solid. LC-MS retention time 2.866 min;
m/z 272.12 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 7.18 (br. s., 1H), 4.59
(br. s., 1 H), 3.36-3.64 (m, 2H), 2.46 (dd, J=13.2, 8.9 Hz, 1H),
2.18-2.31 (m, 1H), 1.64-1.74 (m, 1H), 1.18-1.45 (m, 9H), 0.83 (dt,
J=8.5, 5.8 Hz, 1H), 0.56 (br. s., 1H).
##STR00469##
##STR00470##
Intermediate 123
tert-Butyl
(1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-az-
abicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-
-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0778] Nitrogen was bubbled through a solution of
(1R,3S,5R)-tert-butyl
3-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (400 mg, 0.880 mmol), (1R,3S,5R)-tert-butyl
3-(5-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(313 mg, 1.14 mmol) and Cu(I)I (8.4 mg, 0.044 mmol) in
triethylamine (0.37 mL, 2.6 mmol) and DMF (8 mL) for 10 min. Then
Pd(PPh.sub.3).sub.4 (50.9 mg, 0.044 mmol) was added, nitrogen was
bubbled through the reaction mixture for 1 min, and then the flask
was sealed and heated at 50.degree. C. for 16 h. The reaction was
concentrated to under high vacuum) and the residual solids were
triturated with EtOAc (.about.5 mL) and collected by filtration
(rinsing with EtOAc and hexanes) to yield tert-butyl
(1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3-
.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-a-
zabicyclo[3.1.0]hexane-2-carboxylate (595 mg) as a light yellow
solid. The material was used without further purification. LC-MS
retention time 3.140 min; m/z 647.35 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00471##
Intermediate 124
(1R,3S,5R)-3-(5-(6-((2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hexan-3-yl)-1H-imida-
zol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
ane
[0779] 4M HCl (1.546 mL, 6.18 mmol) in dioxane was added to a
solution of tert-butyl
(1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3-
.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-a-
zabicyclo[3.1.0]hexane-2-carboxylate (200 mg, 0.309 mmol) in
dioxane (3 mL) and the reaction was vigorously stirred for 4 h. The
reaction slurry was concentrated to yield an HCl salt of
(1R,3S,5R)-3-(5-(6-((2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imid-
azol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
xane (177 mg) as a yellow solid. LC-MS retention time 3.403 min;
m/z 893.29 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) d ppm 8.42 (s, 1H), 8.16 (s, 2H), 8.05
(d, J=8.8 Hz, 1H), 8.03 (d, J=8.8 Hz, 1H), 7.97 (dd, J=8.8, 1.8 Hz,
1H), 7.74 (s, 1H), 7.66 (dd, J=8.5, 1.5 Hz, 1H), 5.01 (dd, J=10.8,
7.8 Hz, 1H), 4.78 (dd, J=10.8, 8.3 Hz, 1H), 3.57-3.77 (m, 2H),
3.50-3.56 (m, 1H), 2.61-2.92 (m, 4H), 2.13-2.21 (m, 1H), 2.06-2.13
(m, 1H), 1.25 (ddd, J=7.8, 5.0, 2.5 Hz, 1H), 0.99-1.14 (m, 2H).
##STR00472## ##STR00473##
##STR00474##
Intermediate 125
(1R,3S,5R)-Benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate
[0780] Nitrogen was bubbled through a biphasic solution of
(1R,3S,5R)-benzyl
3-(5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-imidazol--
2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (prepared in an
analogous manner to Intermediate 26) (2.0 g, 4.12 mmol),
(1R,3S,5R)-tert-butyl
3-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (1.872 g, 4.12 mmol) and Na.sub.2CO.sub.3 (2.184 g,
20.60 mmol) in a mixture of EtOH (16.0 mL), toluene (16.0 mL) and
water (16.0 mL) for 15 min. Then, Pd(PPh.sub.3).sub.4 (0.143 g,
0.124 mmol) was added, the reaction was flushed with nitrogen,
sealed and then heated at 95.degree. C. for 10 h. The crude slurry
was diluted with water (.about.20 mL) and extracted with EtOAc
(.about.100 mL). The organic layer was washed with brine, dried
(MgSO.sub.4), filtered and concentrated to a yellow foam. This
material was dissolved into a minimal amount of DCM, and purified
on a BIOTAGE.RTM. Horizon (110 g of SiO.sub.2, 70-85%
EtOAc/hexanes) to yield (1R,3S,5R)-benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (2.257 g) as a light yellow
solidified foam. LC-MS retention time 4.158 min; m/z 733.17 (MH+).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 1 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 5% MeOH/95% H.sub.2O/10
mM ammonium acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.17 (br. s., 1H), 8.07 (s, 1H), 7.86-7.94 (m, 2H),
7.71-7.86 (m, 6H), 7.44 (s, 1H), 7.32 (s, 1H), 7.20 (br. s., 5H),
5.14 (d, J=12.3 Hz, 1H), 4.91-5.05 (m, 1H), 4.63-4.77 (m, 1H),
3.55-3.69 (m, 2H), 3.29-3.34 (m, 1H), 2.46-2.59 (m, 2H), 2.29-2.44
(m, 2H), 1.66-1.80 (m, 2H), 1.30 (br. s., 9H), 0.80-0.93 (m, 2H),
0.61 (br. s., 2H).
##STR00475##
Intermediate 126
(1R,3S,5R)-tert-Butyl
3-(5-(6-(4-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl-
)phenyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-car-
boxylate
[0781] A reaction mixture of (1R,5R)-benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (0.305 g, 0.416 mmol) and
NaHCO.sub.2 (0.070 g, 0.832 mmol) in MeOH (15 mL) was charged with
10% Pd/C (0.022 g, 0.021 mmol). The resulting suspension was vacuum
flushed with N.sub.2 (3.times.) and placed under 1 atm of H.sub.2
(balloon) for 3 h at room temperature. The mixture was then
filtered though a pad of diatomaceous earth (CELITE.RTM.) and
concentrated under vacuum. An off-white solid corresponding to
(1R,3S,5R)-tert-butyl
3-(5-(6-(4-(2-((1R,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)ph-
enyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carbox-
ylate (0.235 g) was recovered and used without further
purification. LC-MS [M+H].sup.+=599; Rt=1.96 min is product. Column
Luna 3u C18 2.times.50 mm; start % B: 0, final % B: 100 Solvent A:
10% Acetonitrile/90% H.sub.2O+1% TFA; Solvent B: 90%
Acetonitrile/10% H.sub.2O+1% TFA; flow rate 4 ml/min. Run time: 5
min. Purity=94%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
12.00 (2 H, br. s.), 8.11-8.32 (3H, m), 7.88-8.02 (5H, m),
7.72-7.88 (9H, m), 7.63 (4H, br. s.), 4.64 (2H, br. s.), 4.42 (1H,
t, J=7.93 Hz), 3.97-4.18 (1H, m), 3.45 (1H, br. s.), 3.17 (3H, s),
2.96 (1H, dd, J=13.43, 6.71 Hz), 2.08-2.43 (6H, m), 1.66 (2H, br.
s.), 1.07-1.50 (20H, m), 0.81-0.89 (3H, m), 0.77 (1H, br. s.),
0.45-0.61 (3H, m), 0.30 (1H, br. s.).
##STR00476##
Intermediate 127
(1R,3S,5R)-tert-Butyl
3-(5-(6-(4-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoy-
l)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)phenyl)naphthalen-2-yl)-
-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0782] HATU (0.164 g, 0.432 mmol) was added to a solution of
(1R,3S,5R)-tert-butyl
3-(5-(6-(4-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl-
)phenyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-car-
boxylate (0.235 g, 0.392 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.069 g, 0.39
mmol) and DIEA (0.137 mL, 0.785 mmol) in DMF (10 mL) and the
resulting yellow solution was stirred at rt overnight. The solvent
was removed under reduced pressure and the residue was dissolved
into methanol, filtered and purified by preparative HPLC (Solvent
A: 10% MeOH/90% water/0.1% TFA; Solvent B: 90% MeOH/10% water/0.1%
TFA; Column. Sunfire Prep MS C18 30.times.100 mm 5u; Wavelength:
220 nM; Flow rate: 30 ml/min; Gradient: 0% B to 100% B over 30 min.
with a 2 min hold time) to yield a TFA salt of
(1R,3S,5R)-tert-butyl
3-(5-(6-(4-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoy-
l)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-
-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (0.10 g)
as a white solid. LC-MS [M+H].sup.+=756; Rt=2.14 min is product.
Column Luna 3u C18 2.times.50 mm; start % B: 0, final % B: 100
Solvent A: 10% Acetonitrile/90% H.sub.2O+1% TFA; Solvent B: 90%
Acetonitrile/10% H.sub.2O+1% TFA; flow rate 4 ml/min. Run time: 5
min. Purity=96%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
14.60 (1 H, br. s.), 8.39 (3H, d, J=4.88 Hz), 8.09-8.24 (4H, m),
7.99-8.08 (6H, m), 7.89-7.98 (5H, m), 7.27 (1H, d, J=8.55 Hz),
4.98-5.07 (1H, m), 4.84 (1H, br. s.), 4.42 (1H, t, J=7.32 Hz), 3.74
(1H, br. s.), 3.55 (4H, s), 2.55 (2H, d, J=9.77 Hz), 2.32-2.44 (3H,
m), 2.13 (1H, dq, J=13.43, 6.71 Hz), 1.94 (1H, dt, J=13.28, 6.79
Hz), 1.75 (1H, dt, J=12.89, 6.22 Hz), 1.40 (4H, d, J=10.07 Hz),
1.11-1.30 (10H, m), 0.90-1.00 (5H, m), 0.73-0.88 (10H, m).
##STR00477##
Intermediate 128
Methyl
(S)-1-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate
[0783] 4N HCl in dioxane (2 mL, 8.00 mmol) was added to a stirred
solution of a TFA salt of (1R,3S,5R)-tert-butyl
3-(5-(6-(4-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoy-
l)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-
-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (0.10 g,
0.12 mmol) in CH.sub.2Cl.sub.2 (20 mL) and the resulting yellow
suspension was stirred at rt for 2 h. The reaction was concentrated
under vacuum and the resulting residue was triturated with
Et.sub.2O to yield an HCl salt of methyl
(S)-1-((1R,3S,5R)-3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-
an-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azab-
icyclo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate HCl (47
mg) as a yellow solid. LC-MS [M+H].sup.+=656; Rt=1.73 min is
product. Column Luna 3u C18 2.times.50 mm; start % B: 0, final % B:
100 Solvent A: 10% Acetonitrile/90% H.sub.2O+1% TFA; Solvent B: 90%
Acetonitrile/10% H.sub.2O+1% TFA; flow rate 4 ml/min. Run time: 5
min. Purity=95%. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. ppm
14.47-15.18 (2H, m), 9.77-10.61 (2H, m), 8.30-8.42 (2H, m), 8.17
(1H, s), 7.93-8.10 (10H, m), 7.27 (1H, d, J=8.55 Hz), 5.08 (1H, t,
J=7.93 Hz), 4.71 (1H, t, J=8.39 Hz), 4.40-4.46 (1H, m), 3.77 (1H,
br. s.), 3.52-3.58 (3H, m), 3.38-3.44 (1H, m), 2.52-2.61 (3H, m),
2.39 (1H, dt, J=13.58, 6.64 Hz), 2.14-2.21 (1H, m), 1.94 (2H, dd,
J=8.24, 4.88 Hz), 1.15 (1H, d, J=5.80 Hz), 0.93 (4H, d, J=6.71 Hz),
0.75-0.89 (6H, m).
##STR00478##
Intermediate 129
(1R,3S,5R)-Benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl-
)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-car-
boxylate
[0784] A solution of 4.0 M HCl (1.167 mL, 4.67 mmol) in dioxane was
added to a stirred solution of (1R,3S,5R)-benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (228 mg, 0.311 mmol) in dioxane (3
mL) and the reaction was stirred vigorously at rt for 3 h. The
reaction was concentrated to yield an HCl salt of (1R,3S,5R)-benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl-
)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-car-
boxylate (233 mg) as a yellow solid. LC-MS retention time 3.988
min; m/z 633.18 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.44 (s, 1H), 8.28 (s, 1H),
8.08-8.18 (m, 3H), 7.92-8.02 (m, 4H), 7.84 (br. s., 3H), 7.31 (br.
s., 5H), 5.21 (d, J=12.1, 1H), 4.95-5.05 (m, 2H), 4.80-4.91 (m,
1H), 3.70-3.77 (m, 1H), 3.59-3.67 (m, 1H), 2.77-2.93 (m, 2H), 2.74
(dd, J=13.4, 9.2 Hz, 1H), 2.46 (dt, J=13.5, 6.7 Hz, 1H), 2.12-2.22
(m, 1H), 1.83-1.96 (m, 1H), 1.27-1.35 (m, 1H), 1.09 (q, J=7.9 Hz,
1H), 0.95 (ddd, J=8.7, 5.9, 5.8 Hz, 1H), 0.75-0.83 (m, 1H).
##STR00479##
Intermediate 130
(1R,3S,5R)-Benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoy-
l)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-
-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0785] HATU (203 mg, 0.533 mmol) was added to a stirred solution of
(1R,5R)-benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl-
)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-car-
boxylate (225 mg) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic
acid (93 mg, 0.53 mmol) in DMF (3 mL) and DIPEA (0.37 mL, 2.1 mmol)
and the reaction was stirred at rt for 3 hr. The crude reaction was
concentrated under a stream of nitrogen and the residue was
dissolved into MeOH and purified by preparative HPLC (MeOH/water
with an ammonium acetate buffer) to yield (1R,5R)-benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoy-
l)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-
-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (200 mg)
as a yellow solid. LC-MS retention time 4.053 min; m/z 790.25
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.28 (s, 2H), 8.13 (d,
J=8.8 Hz, 1H), 8.08 (d, J=8.8 Hz, 1H), 7.94-8.02 (m, 4H), 7.76-7.92
(m, 3H), 7.85 (dd, J=8.7, 1.6 Hz, 1H), 7.29 (br. s., 5H), 5.21 (d,
J=12.1 Hz 1H), 5.16 (dd, J=9.2, 6.9 Hz, 1H), 4.97 (t, J=8.2 Hz,
1H), 4.57 (d, J=6.5 Hz, 1H), 3.78-3.87 (m, 1H), 3.68 (s, 3H),
3.64-3.77 (m, 2H), 2.66-2.77 (m, 2H), 2.40-2.56 (m, 2H), 2.14-2.25
(m, 1H), 2.05-2.14 (m, 1H), 1.87 (d, J=6.5 Hz, 1H), 1.06-1.16 (m,
1H), 1.02 (d, J=6.8 Hz, 3H), 0.94 (d, J=6.8 Hz, 3H), 0.87-0.99 (m,
2H), 0.74-0.82 (m, 1H).
##STR00480##
Intermediate 131
Methyl
(S)-1-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate
[0786] 10% Palladium on carbon (100 mg, 0.094 mmol) was added to a
solution of (1R,5R)-benzyl
3-(5-(4-(6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoy-
l)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-
-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (190 mg,
0.241 mmol) and Na.sub.2CO.sub.3 (50 mg, 0.472 mmol) in THF (10 mL)
and the reaction mixture was vacuum flushed with nitrogen
(3.times.) and then with hydrogen (5.times.). The reaction mixture
was allowed to stir under a balloon of hydrogen for 2 h, filtered
through diatomaceous earth (CELITE.RTM.) and concentrated. The
residue was resubmitted to the reaction conditions (50 mg of 10%
Pd/C used) overnight at rt before being filtered through
diatomaceous earth (CELITE.RTM.) and concentrated. The residue was
dissolved into DMSO/MeOH, filtered and purified by preparative HPLC
(MeOH/H.sub.2O w/0.1% TFA) to yield a TFA salt of methyl
(2S)-1-((1R,3S,5R)-3-(5-(6-(4-(2-((1R,5R)-2-azabicyclo[3.1.0]hexan-3-yl)--
1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (55 mg). LC-MS
retention time 2.147 min; m/z 656.21 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 1 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.28 (s,
1H), 8.24 (s, 1H), 8.12 (d, J=8.9 Hz, 1H), 8.07 (d, J=8.9 Hz, 1H),
7.91-8.01 (m, 4H), 7.83-7.90 (m, 3H), 7.66 (s, 1H), 5.17 (dd,
J=9.2, 7.0 Hz, 1H), 4.67 (dd, J=10.7, 7.6 Hz, 1H), 4.58 (d, J=6.7
Hz, 1H), 3.82-3.87 (m, 1H), 3.69 (s, 3H), 3.44-3.49 (m, 1H),
2.59-2.77 (m, 3H), 2.48-2.56 (m, 1H), 2.16-2.25 (m, 1H), 2.03-2.15
(m, 2H), 1.21 (ddd, J=7.5, 4.9, 2.6 Hz, 1H), 1.09-1.15 (m, 1H),
1.04 (d, J=6.7 Hz, 3H), 0.98-1.03 (m, 1H), 0.95 (d, J=6.7 Hz, 3H),
0.89-0.95 (m, 1H).
##STR00481##
##STR00482##
Intermediate 132
(1R,1'R,3S,3'S,5R,5'R)-tert-Butyl
3,3'-(4,4'-(2,2'-binaphthyl-6,6'-diyl)bis(1H-imidazole-4,2-diyl))bis(2-az-
abicyclo[3.1.0]hexane-2-carboxylate)
[0787] Nitrogen was bubbled a solution of (1R,3S,5R)-tert-butyl
3-(4-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (250 mg, 0.550 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (69.9
mg, 0.275 mmol), 1,1'-bis-(diphenylphosphino)-ferrocene (15.42 mg,
0.028 mmol) and K.sub.2CO.sub.3 (228 mg, 1.651 mmol) in DMSO (12
mL) for 10 min. Then
1,1'-Bis-(diphenylphosphino)-ferrocene)palladium dichloride (22.63
mg, 0.028 mmol) was added to the reaction mixture and the nitrogen
bubbling was continued for 10 min before the reaction was sealed
and then heated at 110.degree. C. for 20 h. The reaction was
partitioned between water (60 mL) and DCM (60 mL) and the organics
were separated, dried (MgSO.sub.4), filtered and concentrated. The
crude residue was purified by BIOTAGE.RTM. Horizon (40 g SiO.sub.2,
70-100% EtOAc/hexanes) to yield (1R,1'R,3S,3'S,5R,5'R)-tert-butyl
3,3'-(4,4'-(2,2'-binaphthyl-6,6'-diyl)bis(1H-imidazole-4,2-diyl))bis(2-az-
abicyclo[3.1.0]hexane-2-carboxylate) (67 mg) as a yellow solid.
LC-MS retention time 4.348 min; m/z 749.29 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.22 (s,
4H), 7.91-8.02 (m, 6 H), 7.87 (d, J=8.5 Hz, 2H), 7.48 (br. s., 2H),
4.68-4.77 (m, 2H), 3.61 (br. s., 2H), 2.56 (dd, J=13.1, 8.8 Hz,
2H), 2.33-2.43 (m, 2H), 1.70-1.79 (m, 2H), 1.31 (br. s., 18H), 0.87
(dt, J=8.5, 5.8 Hz, 2H), 0.63 (br. s., 2H).
##STR00483##
Intermediate 133
6,6'-Bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)-2,-
2'-binaphthyl
[0788] 4.0M HCl (1.0 mL, 4.00 mmol) in dioxane was added to a
stirred solution of (1R,1'R,3S,3'S,5R,5'R)-tert-butyl
3,3'-(4,4'-(2,2'-binaphthyl-6,6'-diyl)bis(1H-imidazole-4,2-diyl))bis(2-az-
abicyclo[3.1.0]hexane-2-carboxylate) (62 mg, 0.083 mmol) in dioxane
(2 mL) and the reaction slurry was stirred vigorously for 3 h. The
slurry was diluted with MeOH and concentrated to dryness to yield
an HCl salt of
6,6'-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)-2-
,2'-binaphthyl (56 mg) as a light yellow solid. LC-MS retention
time 3.971 min; m/z 547.25 (MH-). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.44 (s,
2H), 8.38 (s, 2H), 8.12-8.19 (m, 6H), 8.09 (dd, 2H), 7.97 (dd,
J=8.5, 1.5 Hz, 2H), 4.97 (dd, J=10.5, 8.1 Hz, 2H), 3.62-3.66 (m,
2H), 2.79-2.89 (m, 4H), 2.15-2.22 (m, 2H), 1.29-1.34 (m, 2H), 1.11
(q, J=7.9 Hz, 2H).
##STR00484##
##STR00485##
Intermediate 134
6-Bromo-2-(1-ethoxyvinyl)quinoline
[0789] Dichlorobis(triphenylphosphine)-palladium(II) (0.289 g,
0.412 mmol) was added to a solution of 6-bromo-2-chloroquinoline
(1.0 g, 4.1 mmol) and tributyl(1-ethoxyvinyl)stannane (1.79 g, 4.95
mmol) in dioxane (8 mL) and the mixture was stirred at 100.degree.
C. for 5 h, desired product was identified by LC-MS. The crude
reaction mixture was concentrated and purified by flash silica gel
chromatography (eluted with 1:1 hexanes/DCM) to yield product
6-bromo-2-(1-ethoxyvinyl)quinoline (720 mg) as white solid. LC-MS
retention time 4.091 min; m/z 279.84 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00486##
Intermediate 135
2-Bromo-1-(6-bromoquinolin-2-yl)ethanone
[0790] NBS (407 mg, 2.287 mmol) was added to a solution of
6-bromo-2-(1-ethoxyvinyl)quinoline (530 mg, 1.91 mmol) in THF (10
mL) and water (2.5 mL) and the mixture was stirred at rt for 2 h.
The reaction mixture was partitioned between EtOAc and brine and
the organic layer was concentrated. The crude material was purified
by flash silica gel chromatography (eluted with Et.sub.2O/hexanes,
gradient from 0 to 5% Et.sub.2O) to yield
2-bromo-1-(6-bromoquinolin-2-yl)ethanone (380 mg) as white solid.
LC-MS retention time 3.988 min; m/z 329.84 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00487##
Intermediate 136
(1R,3S,5R)-3-(2-(6-Bromoquinolin-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate
[0791] DIPEA (0.21 mL, 1.2 mmol) was added to a solution of
2-bromo-1-(6-bromoquinolin-2-yl)ethanone (268 mg, 0.815 mmol)) and
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (204 mg, 0.896 mmol) in acetonitrile (8 mL) and the reaction
mixture was stirred at rt for 16 h. The reaction mixture was
diluted with sat. aq. NaHCO.sub.3 (5 mL) and extracted with EtOAc
(2.times.20 mL). The combined organic layers were washed with
brine, dried (MgSO.sub.4), filtered and concentrated. The crude
product was purified by flash silica gel chromatography (used DCM
as loading solvent, eluted with Et.sub.2O/hexanes, gradient from
10% to 30% Et.sub.2O) to yield
(1R,3S,5R)-3-(2-(6-bromoquinolin-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (332 mg) as white
solid. LC-MS retention time 4.283 min; m/z 476.88 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00488##
Intermediate 137
tert-Butyl
(1R,3S,5R)-3-(4-(6-bromo-2-quinolinyl)-1H-imidazol-2-yl)-2-azab-
icyclo[3.1.0]hexane-2-carboxylate
[0792] Ammonium acetate (990 mg, 12.8 mmol) was added to a solution
of (1R,3S,5R)-3-(2-(6-bromoquinolin-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (407 mg, 0.856 mmol) in
toluene (10 mL) and the mixture was stirred at 120.degree. C. for 3
h. The reaction mixture was diluted with sat. aq NaHCO.sub.3 (10
mL) and extracted with EtOAc (50 mL). The organic layer was washed
with brine, dried (MgSO.sub.4), filtered and concentrated. The
crude material was purified by flash silica gel chromatography
(eluted with 1:1 EtOAc/hexanes) to yield tert-butyl
(1R,3S,5R)-3-(4-(6-bromo-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1-
.0]hexane-2-carboxylate (272 mg) as light yellow solid. LC-MS
retention time 3.306 min; m/z 456.99 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, chloroform-d) .delta. ppm
8.08 (d, J=8.5 Hz, 1H), 7.95 (d, J=2.2 Hz, 1H), 7.78 (dd, J=8.9,
2.2 Hz, 1H), 7.73-8.14 (m, 3H), 4.86-4.98 (m, 1H), 3.70-3.82 (m,
0.5H), 3.26-3.39 (m, 1H), 2.46 (dd, J=13.3, 9.3 Hz, 1H), 1.84-1.91
(m, 0.5H), 1.74-1.84 (m, 1H), 1.52 (s, 9H), 0.87-0.97 (m, 1H),
0.48-0.61 (m, 1H).
##STR00489##
Intermediate 138
tert-Butyl
(1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2--
azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)-2-quinolinyl)-1H-imida-
zol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0793] Pd(OAc).sub.2 (7.40 mg, 0.033 mmol) was added to a solution
of tert-butyl
(1R,3S,5R)-3-(4-(6-bromo-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1-
.0]hexane-2-carboxylate (150 mg, 0.329 mmol), (1R,3S,5R)-tert-butyl
3-(5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-imidazol--
2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (149 mg, 0.329 mmol),
dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine (27.0 mg, 0.066
mmol) and K.sub.2CO.sub.3 (137 mg, 0.988 mmol) in THF (2 mL) and
water (0.50 mL) and the reaction mixture was stirred at 110.degree.
C. for 2 h. The reaction mixture was diluted with MeOH, filtered
and purified by preparative HPLC (H.sub.2O-MeOH with 10 mM
NH.sub.4OAc buffer) to yield tert-butyl
(1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)--
2-azabicyclo[3.1.0]hexane-2-carboxylate (140 mg) as bright yellow
solid. LC-MS retention time 3.150 min; m/z 700.36 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode.
##STR00490##
Intermediate 139
2-(2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-6-(4-(2-((-
1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)quinoline
[0794] A solution of 4M HCl (1 mL, 4.00 mmol) in dioxane was added
to a suspension of tert-butyl
(1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)--
2-azabicyclo[3.1.0]hexane-2-carboxylate (130 mg, 0.186 mmol) in
dioxane (3 mL), MeOH (0.5 mL) and DCM (3 mL) and the mixture was
stirred at rt for 2 h. The reaction was concentrated to yield an
HCl salt of
2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-6-(4-(2-(-
(1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)quinoline
(140 mg) as yellow solid. LC-MS retention time 2.063 min; m/z
500.37 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.09 (d, J=9.0 Hz,
1H), 8.66 (s, 1H), 8.56-8.62 (m, 2H), 8.47-8.53 (m, 2H), 8.07 (s,
1H), 8.02-8.06 (m, 4H), 4.97 (dd, J=10.8, 8.0 Hz, 1H), 3.65-3.71
(m, 1H), 3.55-3.65 (m, 2H), 2.73-2.86 (m, 3H), 2.61-2.72 (m, 1H),
2.07-2.20 (m, 2H), 1.25-1.35 (m, 2H), 1.07 (t, 2H).
##STR00491##
##STR00492##
Intermediate 140
1-(4-(5-Acetylnaphthalen-1-yl)phenyl)ethanone
[0795] A solution of sodium carbonate (2.55 g, 24.09 mmol) in water
(20 mL) was added to a solution of
1-(5-bromonaphthalen-1-yl)ethanone (1.2 g, 4.8 mmol) and
4-acetylphenylboronic acid (1.03 g, 6.26 mmol) in toluene (20 mL)
and ethanol (20 mL) and then heterogeneous solution was vigorously
stirred with bubbling nitrogen for 15 min. Then Pd(PPh.sub.3).sub.4
(0.111 g, 0.096 mmol) was added and the reaction vessel was sealed
and heated at reflux for 6 h. The reaction was cooled and
concentrated to dryness. The crude residue was taken into EtOAc
(.about.150 mL) and water (.about.100 mL). The layers were
separated and the organic layer was washed with brine, dried
(MgSO.sub.4), filtered and concentrated to an orange solid. The
orange solid was purified on a BIOTAGE.RTM. Horizon (40 g
SiO.sub.2, DCM) to yield
1-(4-(5-acetylnaphthalen-1-yl)phenyl)ethanone (1.31 g) as an
off-white solid. LC-MS retention time 3.736 min; m/z 289.19 (MH+).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 8.75 (d, J=8.8 Hz,
1H), 8.11 (d, J=8.5 Hz, 2H), 7.99 (d, J=8.5 Hz, 1H), 7.94 (dd,
J=7.2, 1.1 Hz, 1H), 7.67 (dd, J=8.5, 7.0 Hz, 1H), 7.58 (d, J=8.3
Hz, 2H), 7.45-7.51 (m, 2H), 2.79 (s, 3H), 2.70 (s, 3H).
##STR00493##
Intermediate 141
2-Bromo-1-(4-(5-(2-bromoacetyl)naphthalen-1-yl)phenyl)ethanone
[0796] A solution of bromine (0.47 mL, 9.1 mmol) in DCM (10 mL) was
added to a solution of
1-(4-(5-acetylnaphthalen-1-yl)phenyl)ethanone (1.285 g, 4.46 mmol)
in DCM (20 mL) and the reaction was stirred at rt for 1 d. The
reaction was concentrated to a viscous light orange oil
(.about.1.92 g). By .sup.1H NMR the major product appears to be the
desired product with peaks corresponding to the two mono brominated
regioisomers (.about.10%). The material was used without
purification. .sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 8.64
(d, J=8.5 Hz, 1H), 8.13-8.17 (m, 2H), 8.05 (d, J=8.5 Hz, 1H), 7.94
(dd, J=7.2, 1.1 Hz, 1H), 7.71 (dd, J=8.8, 7.0 Hz, 1H), 7.60-7.64
(m, 2H), 7.48-7.55 (m, 2 H), 4.60 (s, 2H), 4.53 (s, 2H).
##STR00494##
Intermediate 142
(1R,3S,5R)-3-(2-(4-(5-(2-((1R,3S,5R)-2-(tert-Butoxycarbonyl)-2-azabicyclo[-
3.1.0]hexane-3-carbonyloxy)acetyl)naphthalen-1-yl)phenyl)-2-oxoethyl)
2-tert-butyl 2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate
[0797] DIPEA (2.337 mL, 13.38 mmol) was added to a solution of
crude
2-bromo-1-(4-(5-(2-bromoacetyl)naphthalen-1-yl)phenyl)ethanone
(1.99 g, 4.46 mmol) and
(1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (2.03 g, 8.92 mmol) in acetonitrile (50 mL) and the clear
yellow solution was stirred at rt overnight. The reaction was
concentrated to a yellow solidified foam which was purified on a
BIOTAGE.RTM. Horizon (SiO.sub.2, EtOAc/hexanes) to yield
(1R,3S,5R)-3-(2-(4-(5-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hexane-3-carbonyloxy)acetyl)naphthalen-1-yl)phenyl)-2-oxoethyl)2-te-
rt-butyl 2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (2.95 g) as a
white fluffy solid. LC-MS retention time 4.363 min; m/z 737.38
(MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and Solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.57 (d, J=8.8 Hz,
1H), 8.14 (d, J=8.0 Hz, 2H), 7.99-8.06 (m, 2H), 7.68 (dd, J=8.8,
7.0 Hz, 1H), 7.64 (d, J=8.3 Hz, 2H), 7.52-7.60 (m, 2H), 5.60-5.70
(m, 1H), 5.32-5.56 (m, 3H), 4.24-4.34 (m, 2H), 3.38-3.49 (m, 2H),
2.59-2.69 (m, 1H), 2.38-2.55 (m, 3H), 1.61-1.77 (m, 2H), 1.47 (br.
s., 18H), 0.81-0.94 (m, 2H), 0.50-0.58 (m, 2H).
##STR00495##
Intermediate 143
(1R,3S,5R)-tert-Butyl
3-(5-(4-(5-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate
[0798] A solution of
(1R,3S,5R)-3-(2-(4-(5-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hexane-3-carbonyloxy)acetyl)naphthalen-1-yl)phenyl)-2-oxoethyl)
2-tert-butyl 2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate (2.01 g,
2.72 mmol) and ammonium acetate (4.19 g, 54.4 mmol) in toluene (40
mL) was stirred and heated at 100.degree. C. overnight. The
reaction was concentrated to dryness and the brown solids were
partitioned between DCM (150 mL) and 1/2 sat. aq. NaHCO.sub.3 (100
mL). The organics were dried (MgSO.sub.4), filtered and
concentrated. The crude material was not easily purified by flash
column chromatography so all fractions containing the desired
product or starting material were collected, concentrated and
resubmitted to the reaction conditions (now 110.degree. C.
overnight). The reaction was concentrated and the brown solids were
partitioned between DCM (150 mL) and 1/2 sat NaHCO.sub.3 (100 mL).
The organics were dried (MgSO.sub.4), filtered and concentrated.
Approximately .about.30-35% of the material was purified by
preparative HPLC (dissolved into MeOH, filtered, 80-100%
MeOH/water, ammonium acetate buffer) to yield (1R,3S,5R)-tert-butyl
3-(5-(4-(5-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (148 mg) as a yellow solid. The
remaining material was purified by preparative HPLC (dissolved into
MeOH, filtered, 60-100% MeOH/water, TFA buffer) to yield a TFA salt
(1R,3S,5R)-tert-butyl
3-(5-(4-(5-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (299 mg). LC-MS retention time
3.100 min; m/z 699.56 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.29 (d, J=8.5 Hz,
1H), 7.90 (d, J=8.5 Hz, 1H), 7.83 (d, J=8.3 Hz, 2H), 7.59 (dd,
J=7.0, 1.0 Hz, 1H), 7.53 (dd, J=8.5, 7.0 Hz, 1H), 7.43-7.50 (m,
4H), 7.42 (s, 1H), 7.23 (s, 1H), 4.66-4.80 (m, 2H), 3.60 (br. s.,
2H), 2.56 (dt, J=13.1, 8.9 Hz, 2H), 2.32-2.46 (m, 2H), 1.70-1.78
(m, 2H), 1.38 (br. s., 18H), 0.82-0.90 (m, 2H), 0.63 (br. s.,
2H).
##STR00496##
Intermediate 144
(1R,3S,5R)-3-(5-(4-(5-(2-((1R,3S,5R)-2-Azabicyclo[3.1.0]hexan-3-yl)-1H-imi-
dazol-5-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
xane
[0799] 4M HCl (0.757 mL, 3.03 mmol) in dioxane was added to a
solution of (1R,3S,5R)-tert-butyl
3-(5-(4-(5-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexa-
n-3-yl)-1H-imidazol-5-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexane-2-carboxylate (141 mg, 0.202 mmol) in dioxane
(3.0 mL) and the reaction was stirred at rt for 6 h. The solids
were washed down the sides of the vial with MeOH and the reaction
was concentrated under a stream nitrogen to yield an HCl salt of
(1R,3S,5R)-3-(5-(4-(5-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-im-
idazol-4-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
exane (119 mg) as a yellow solid. LC-MS retention time 2.333 min;
m/z 499.57 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.15 (d, J=8.3 Hz,
1H), 8.03-8.08 (m, 2H), 8.01 (d, J=8.0 Hz, 2H), 7.95 (s, 1H), 7.77
(d, J=6.5 Hz, 1H), 7.68-7.74 (m, 1H), 7.65 (d, J=8.0 Hz, 2H),
7.57-7.64 (m, 2H), 5.03 (t, J=9.4 Hz, 1H), 4.97 (dd, J=10.8, 8.0
Hz, 1H), 3.57-3.66 (m, 2H), 2.76-2.89 (m, 4H), 2.11-2.21 (m, 2H),
1.27-1.34 (m, J=7.7, 5.1, 5.1, 2.6 Hz, 2H), 1.05-1.14 (m, 2H).
##STR00497##
##STR00498##
Intermediate 145
(2S,3aS,6aS)-2-(2-(4-(6-(2-((2S,3aS,6aS)-1-(tert-Butoxycarbonyl)-octahydro-
cyclopenta[b]pyrrole-2-carbonyloxy)acetyl)naphthalen-2-yl)phenyl)-2-oxoeth-
yl)1-tert-butyl
hexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate
[0800] Neat DIPEA (0.068 mL, 0.388 mmol) was added to a stirred
suspension of
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (79
mg, 0.176 mmol) and
(2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carbo-
xylic acid (90 mg, 0.353 mmol) in an. acetonitrile (1.5 mL) and
chloroform (1.5 mL) and the mixture was stirred at r.t. overnight.
Reaction mixture was evaporated to dryness and then purified by
silica gel FCC (3% MeOH in DCM) to afford the Intermediate 145 as a
tan solid. LC-MS retention time: 2.480 min; m/z 793.7
(M-H).sup.-.
##STR00499##
Intermediate 146
(2S,3aS,6aS)-tert-Butyl
2-(4-(4-(6-(2-((2S,3aS,6aS)-1-(tert-butoxycarbonyl)-octahydrocyclopenta[b-
]pyrrol-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)he-
xahydrocyclopenta[b]pyrrole-1(2H)-carboxylate
[0801] A stirred suspension of
(2S,3aS,6aS)-2-(2-(4-(6-(2-((2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydro-
cyclopenta[b]pyrrole-2-carbonyloxy)acetyl)naphthalen-2-yl)phenyl)-2-oxoeth-
yl)1-tert-butyl hexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate
(140 mg, 0.176 mmol) and ammonium acetate (272 mg, 3.52 mmol) in
xylene (4 mL) was heated at 140.degree. C. for 2 h. Reaction
mixture was cooled to r.t. and diluted with EtOAc (20 ml) and
washed with satd. NaHCO.sub.3, water, brine and dried
(Na.sub.2SO.sub.4) to afford a brown solid which was purified by
silica gel FCC (3-5% MeOH in DCM) to afford the
(2S,3aS,6aS)-tert-butyl
2-(4-(4-(6-(2-((2S,3aS,6aS)-1-(tert-butoxycarbonyl)-octahydrocyclopenta[b-
]pyrrol-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)he-
xahydrocyclopenta[b]pyrrole-1(2H)-carboxylate. LC-MS retention
time: 2.577 min; m/z 755.7 (M+H).sup.+.
##STR00500##
Intermediate 147
(2S,3aS,6aS)-2-(4-(4-(6-(2-((2S,3aS,6aS)-Octahydrocyclopenta[b]pyrrol-2-yl-
)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)octahydrocyclo-
penta[b]pyrrole
[0802] 4N HCl in dioxane (20 equiv) was added to a solution of
(2S,3aS,6aS)-tert-butyl
2-(4-(4-(6-(2-((2S,3aS,6aS)-1-(tert-butoxycarbonyl)-octahydrocyclopenta[b-
]pyrrol-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)he-
xahydrocyclopenta[b]pyrrole-1(2H)-carboxylate in an. DCM and the
mixture was stirred at rt for 2-3 h. Anhydrous toluene was added to
the resultant yellow suspension and then evaporated to dryness to
an HCl salt of
(2S,3aS,6aS)-2-(4-(4-(6-(2-((2S,3aS,6aS)-octahydrocyclopenta[b]pyrrol-2-y-
l)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)octahydrocycl-
openta[b]pyrrole as a beige solid: LC-MS retention time: 2.343 min;
m/z 555 (M+H).sup.+.
##STR00501##
##STR00502##
Intermediate 148
(1R,3S,5R)-3-(4-(6-Bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexane
[0803] TFA (1.028 mL, 13.34 mmol) was added to a solution of
(1R,3S,5R)-tert-butyl
3-(4-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (Intermediate 120) (606 mg, 1.334 mmol) in DCE (20
mL) and reaction was stirred at rt for 2 h. Then additional TFA (1
mL) was added and the reaction was stirred at rt for 4 h. The
reaction was concentrated to dryness and the crude brown oil was
dissolved into diethyl ether and concentrated under vacuum
(2.times.) to yield a TFA salt of
(1R,3S,5R)-3-(4-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hexane (786 mg) as a yellow solid. The material was used
without further purification. LC-MS retention time 2.558 min; m/z
352.03, 354.05 (1:1) (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 10.55 (br. s., 1H),
8.02 (s, 1H), 7.93 (d, J=1.5 Hz, 1H), 7.72 (d, J=8.5 Hz, 1H), 7.65
(d, J=8.8 Hz. 1H), 7.59 (dd, J=8.8, 2.0 Hz, 1H), 7.52 (dd, J=8.7,
1.6 Hz, 1H), 7.44 (s, 1H), 5.10 (dd, J=11.8, 7.3 Hz, 1H), 3.49-3.54
(m, 1H), 2.94 (td, J=12.7, 4.6 Hz, 1H), 2.68 (dd, J=13.2, 7.2 Hz,
1H), 2.06-2.14 (m, 1H), 1.20-1.26 (m, 1H), 1.00-1.09 (m, 1H).
##STR00503##
Intermediate 149
Methyl
(S)-1-((1R,3S,5R)-3-(4-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-
-azabicyclo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate
[0804] HATU (411 mg, 1.082 mmol) was added to a solution of a TFA
salt of
(1R,3S,5R)-3-(4-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hexane (525 mg, 0.902 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (190 mg, 1.08
mmol) in DMF (10 mL) and DIPEA (0.79 mL, 4.5 mmol) and the reaction
was stirred at rt for 4 h. The reaction was diluted with EtOAc (-80
mL) and washed with sat. aq. NaHCO.sub.3, water (30 mL) and brine
(30 mL). The organics were then dried (MgSO.sub.4), filtered and
concentrated to a brown oil (625 mg). This material was purified on
a BIOTAGE.RTM. Horizon (40 g SiO.sub.2, 40-80% EtOAc/hexanes) to
yield methyl
(S)-1-((1R,3S,5R)-3-(4-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (412 mg)
as a yellow glass. LC-MS retention time 1.843 min; m/z 511.24,
513.12 (1:1) (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1.0 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold
time of 1 min, and an analysis time of 3 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00504##
Intermediate 150
(1R,3S,5R)-tert-Butyl
3-(4-((6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)ethynyl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0805] Nitrogen was bubbled through a mixture of
(1R,3S,5R)-tert-butyl
3-(5-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(Intermediate 122) (118 mg, 0.432 mmol), methyl
(S)-1-((1R,3S,5R)-3-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (170 mg,
0.332 mmol) and CuI (3.17 mg, 0.017 mmol) in DMF (4 mL) and TEA
(0.14 mL, 0.10 mmol) for 15 min. Then Pd(PPh.sub.3).sub.4 (19.2 mg,
0.017 mmol) was added and the reaction vessel was sealed and heated
at 60.degree. C. overnight. The reaction was cooled and then
additional CuI (5 mg) and TEA (100 .mu.L) were added to the
reaction and nitrogen was bubbled through the reaction for 15 min.
Then additional Pd(PPh.sub.3).sub.4 (12 mg) was added and the
reaction was sealed and stirred at 70.degree. C. for 24 h. The
reaction was concentrated under high vacuum and the residue was
partitioned between DCM (25 mL) and water (25 mL). The aqueous was
extracted with DCM (2.times.10 mL) and the combined organics were
washed with brine (.about.20 mL), dried (MgSO.sub.4), filtered and
concentrated to a viscous dark oil. The material was dissolved into
MeOH, filtered and purified by preparative HPLC (MeOH/water,
ammonium acetate buffer) to yield (1R,3S,5R)-tert-butyl
3-(4-((6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)ethynyl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (187.4
mg) as a yellow solid. The material was used without further
purification. LC-MS retention time 1.720 min; m/z 704.59 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 1.0 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00505##
##STR00506##
Intermediate 151
(2S,5S)-2-(2-(6-Bromonaphthalen-2-yl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate
[0806] DIPEA (0.823 mL, 4.71 mmol) was added to a stirred slurry of
2-bromo-1-(6-bromonaphthalen-2-yl)ethanone (Intermediate 118) (1.03
g, 3.14 mmol) and
(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic
acid (0.720 g, 3.14 mmol) in acetonitrile (50 mL) and the reaction
was stirred at rt overnight. The reaction was concentrated to
dryness and purified by BIOTAGE.RTM. Horizon (40 g SiO.sub.2,
10-20% EtOAc/hexanes) to yield
(2S,5S)-2-(2-(6-bromonaphthalen-2-yl)-2-oxoethyl) 1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate (1.09 g) as a yellow
solidified foam. LC-MS retention time 4.413 min; m/z 476, 478.20
(1:1) (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold time
of 1 min, and an analysis time of 5 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.57 (br. s., 1H), 8.14 (d,
J=1.5 Hz, 1H), 8.01 (d, J=8.5 Hz, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.91
(d, J=8.8 Hz, 1H), 7.68 (dd, J=8.8, 1.8 Hz, 1H), 5.62-5.70 (m, 1H),
5.47-5.60 (m, 1H), 4.46 (t, J=7.3 Hz, 1H), 3.92-4.06 (m, 1H),
2.26-2.37 (m, 2H), 2.05-2.19 (m, 1H), 1.67-1.82 (m, 1H), 1.46 (d,
J=6.8 Hz, 9H), 1.30 (d, J=6.5 Hz, 3H).
##STR00507##
Intermediate 152
(2S,5S)-tert-Butyl
2-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-5-methylpyrrolidine-1-car-
boxylate
[0807] Ammonium acetate (3.79 g, 49.1 mmol) was added to a solution
of (2S,5S)-2-(2-(6-bromonaphthalen-2-yl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate (1.17 g, 2.47 mmol) in
toluene (30 mL) and the reaction was placed into an oil bath which
had been preheated to 100.degree. C. and stirred at that
temperature overnight. The reaction was allowed to cool to rt,
concentrated and partitioned between DCM (100 mL) and 1/2 sat. aq.
NaHCO.sub.3 (75 mL). The organics layer was washed with brine (-50
mL), dried (MgSO.sub.4), filtered and concentrated to a solidified
tan foam. This material was purified on a BIOTAGE.RTM. Horizon (160
g SiO.sub.2, loaded with DCM, 20-35% EtOAc/hexanes) to yield
(2S,55)-tert-butyl
2-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-5-methylpyrrolidine-1-car-
boxylate (893 mg) as a solidified tan foam. LC-MS retention time
3.410 min; m/z 456, 458.22 (1:1) (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min, and an analysis time of 4 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.19
(br. s., 1H), 8.01 (s, 1H), 7.83-7.93 (m, 1H), 7.76-7.83 (m, 2H),
7.55 (dd, J=8.7, 1.6 Hz, 1H), 7.47 (br. s., 1H), 4.94 (br. s., 1H),
3.99-4.09 (m, 1H), 2.22-2.36 (m, 2H), 2.09-2.21 (m, 1H), 1.72 (br.
s., 1H), 1.42 (d, J=6.0 Hz, 3H), 1.34 (br. s., 9H).
##STR00508##
Intermediate 153
(1R,3S,5R)-tert-Butyl
3-(4-((6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl)-1H--
imidazol-4-yl)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate
[0808] Nitrogen was bubbled through a solution of
(2S,5S)-tert-butyl
2-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-5-methylpyrrolidine-1-car-
boxylate (127 mg, 0.277 mmol), (1R,3S,5R)-tert-butyl
3-(5-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(91 mg, 0.33 mmol) and CuI (2.6 mg, 0.014 mmol) in DMF (3 mL) and
TEA (0.12 mL, 0.83 mmol) for 15 min. Then Pd(PPh.sub.3).sub.4 (16
mg, 0.014 mmol) was added and the reaction vessel was flushed with
nitrogen, sealed and heated at 50.degree. C. overnight. The
reaction was cooled to rt, nitrogen was bubbled through the
reaction mixture for 20 min., and then additional CuI (.about.4 mg)
and Pd(PPh.sub.3).sub.4 (15 mg) were added. The reaction was
flushed with nitrogen for 5 min., sealed and heated at 60.degree.
C. for 1 d. The reaction was cooled to rt, diluted with methanol,
filtered and purified by preparative HPLC (MeOH/water with TFA
buffer) to yield a TFA salt of (1R,3S,5R)-tert-butyl
3-(5-((6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl)-1H--
imidazol-5-yl)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate (56.7 mg) as a yellow solid. LC-MS retention
time 3.161 min; m/z 649.62 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.31 (s,
1H), 8.21 (s, 1H), 8.07 (d, J=8.8 Hz, 1H), 7.99-8.04 (m, 2H), 7.91
(dd, J=8.8, 1.8 Hz, 1H), 7.77-7.87 (m, 1H), 7.69 (dd, J=8.7, 1.4
Hz, 1H), 5.13 (t, J=7.5 Hz, 1H), 4.12 (br. s., 1H), 3.61 (br. s.,
1H), 2.66 (br. s., 1H), 2.45-2.54 (m, 1H), 2.33-2.42 (m, 1H),
2.22-2.32 (m, 2H), 1.75-1.86 (m, 2H), 1.25-1.59 (m, 21H), 1.06
(none, 1H), 0.90 (dt, J=8.5, 5.8 Hz, 1H), 0.71 (br. s., 1H).
##STR00509##
Intermediate 154
(1R,3S,5R)-3-(4-((6-(2-((2S,5S)-5-Methylpyrrolidin-2-yl)-1H-imidazol-4-yl)-
naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
[0809] 4M HCl (0.611 mL, 2.445 mmol) in dioxane was added to a
mixture of a TFA salt of (1R,3S,5R)-tert-butyl
3-(5-((6-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl)-1H--
imidazol-5-yl)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate (53.6 mg, 0.061 mmol) in dioxane (1.5 mL)
and the reaction was vigorously stirred for 2 h. The reaction was
concentrated under a steam of nitrogen overnight to yield an HCl
salt of
(1R,3S,5R)-3-(4-((6-(2-((2S,5S)-5-methylpyrrolidin-2-yl)-1H-imidazol-4-yl-
)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(37 mg) which was used without further purification. LC-MS
retention time 2.302 min; m/z 449.40 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00510##
##STR00511##
Intermediate 155
(1R,3S,5R)-3-(4-((Trimethylsilyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hexane
[0810] TFA (500 .mu.l, 6.49 mmol) was added to a solution of
(1R,3S,5R)-tert-butyl
3-(5-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-
e-2-carboxylate (299 mg, 0.865 mmol) in DCE (5 mL) and the reaction
was stirred at rt for 30 min. The reaction was concentrated to
dryness and then resubmitted to the reaction conditions for 5 h.
The reaction was concentrated to yield a TFA salt of
(1R,3S,5R)-3-(4-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[-
3.1.0]hexane as a viscous brown oil which was used without further
purification. LC-MS retention time 2.755 min; m/z 246.25 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00512##
Intermediate 156
Methyl
(S)-3-methyl-1-oxo-1-((1R,3S,5R)-3-(4-((trimethylsilyl)ethynyl)-1H--
imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-2-yl)butan-2-ylcarbamate
[0811] HATU (428 mg, 1.125 mmol) was added to a stirred solution of
(1R,3S,5R)-3-(5-((trimethylsilyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[-
3.1.0]hexane (212 mg, 0.865 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (197 mg, 1.125
mmol) in DMF (3 mL) and DIPEA (1.2 mL, 6.9 mmol) and the reaction
was stirred at rt overnight. The reaction was concentrated to
dryness and then purified with a BIOTAGE.RTM. Horizon (25 g
SiO.sub.2, 40-60% EtOAc/hexanes) to yield methyl
(S)-3-methyl-1-oxo-1-((1R,3S,5R)-3-(5-((trimethylsilyl)ethynyl)-1H-imidaz-
ol-2-yl)-2-azabicyclo[3.1.0]hexan-2-yl)butan-2-ylcarbamate (203 mg)
as a yellow glass. LC-MS retention time 3.095 min; m/z 403.29
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 7.17 (br. s., 1H),
5.04-5.11 (m, 1H), 4.54 (d, J=6.8 Hz, 1H), 3.65 (s, 3H), 3.54-3.61
(m, 1H), 2.82 (s, 1H), 2.39-2.50 (m, 1H), 2.27-2.36 (m, 1H),
2.04-2.15 (m, 1H), 1.94-2.04 (m, 1H), 1.11 (dt, J=8.7, 5.6 Hz, 1H),
0.95 (d, J=6.8 Hz, 3H), 0.90 (d, J=6.8 Hz, 3H), 0.73-0.79 (m, 1H),
0.21 (s, 9H).
##STR00513##
Intermediate 157
Methyl
(S)-1-((1R,3S,5R)-3-(4-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0-
]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate
[0812] Potassium carbonate (34.3 mg, 0.248 mmol) was added to a
solution of methyl
(S)-3-methyl-1-oxo-1-((1R,3S,5R)-3-(5-((trimethylsilyl)ethynyl)-
-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-2-yl)butan-2-ylcarbamate
(200 mg, 0.497 mmol) in methanol (5 mL) and the reaction was
flushed with nitrogen, sealed and then heated at 50.degree. C. for
4 h. and then stirred at rt overnight. The reaction was
concentrated to dryness and then purified on a BIOTAGE.RTM. Horizon
(12 g SiO.sub.2, 70-90% EtOAc/hexanes) to yield methyl
(S)-1-((1R,3S,5R)-3-(5-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-
-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (151 mg) as a yellow
glass. LC-MS retention time 1.788 min; m/z 331.32 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00514##
Intermediate 158
(1R,3S,5R)-tert-Butyl
3-(5-(6-((2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)ethynyl)naphthalen-2-yl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0813] A mixture of (1R,3S,5R)-tert-butyl
3-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-
-2-carboxylate (208 mg, 0.457 mmol), methyl
(S)-1-((1R,3S,5R)-3-(5-ethynyl-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexan-
-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (151 mg, 0.457 mmol) and
CuI (8.70 mg, 0.046 mmol) in DMF (4 mL) and TEA (0.19 mL, 1.4 mmol)
was vacuum flushed with nitrogen (6.times.) over 10 minutes. Then
Pd(PPh.sub.3).sub.4 (53 mg, 0.046 mmol) was added and the reaction
mixture was vacuum flushed with nitrogen (3.times.), sealed and
heated at 60.degree. C. for 20 h. The reaction was concentrated
under a steam of nitrogen overnight and then purified preparative
HPLC (40-80% MeOH/water with a TFA buffer) to yield a TFA salt of
(1R,3S,5R)-tert-butyl
3-(5-(6-((2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)ethynyl)naphthalen-2-yl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (270 mg)
as a yellow solid. LC-MS retention time 2.995 min; m/z 704.70
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.28 (s, 1H), 8.18
(s, 1H), 8.06 (d, J=8.8 Hz, 1H), 7.98-8.03 (m, 2H), 7.89 (dd,
J=8.5, 1.8 Hz, 1H), 7.69-7.85 (m, 1 H), 7.67 (dd, J=8.7, 1.4 Hz,
1H), 5.11 (br. s., 1H), 4.84-4.91 (m, 2H), 4.55 (d, J=5.8 Hz, 1H),
3.70-3.77 (m, 1H), 3.67 (s, 3H), 2.72 (dd, J=13.6, 9.0 Hz, 1H),
2.55-2.63 (m, 1H), 2.39-2.50 (m, 2H), 2.11-2.20 (m, 1H), 2.02-2.11
(m, 1H), 1.81-1.89 (m, 1H), 1.45 (br. s., 9H), 1.07-1.14 (m, 1H),
1.00 (d, J=6.8 Hz, 3H), 0.93 (d, J=6.8 Hz, 3H), 0.84-0.93 (m, 2H),
0.74 (br. s., 1H).
##STR00515##
##STR00516##
Intermediate 159
1-(6-Bromoquinolin-2-yl)ethanone
[0814] 3M HCl (aq.) (10 mL, 30.0 mmol) was added to a suspension of
6-bromo-2-(1-ethoxyvinyl)quinoline (780 mg, 2.80 mmol) in acetone
(20 mL) and the reaction mixture was stirred at rt for 5 h. The
reaction mixture was concentrated, dissolved into THF (20 mL) and
treated with 3N HCl(aq.) (5 mL) was clear reaction solution was
stirred at 60.degree. C. for 5 h. The reaction was, cooled,
neutralized with aq. NaOH and NaHCO.sub.3 and extracted with EtOAc.
The organic layer was dried, and concentrated to yield crude
1-(6-bromoquinolin-2-yl)ethanone (710 mg) as white solid. LC-MS
retention time 3.703 min; m/z 250, 251.99 (1:1) (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00517##
Intermediate 160
1-(6-(4-Acetylphenyl)quinolin-2-yl)ethanone
[0815] Pd(OAc).sub.2 (62.9 mg, 0.280 mmol) was added to a solution
of 4-acetylphenylboronic acid (689 mg, 4.20 mmol),
1-(6-bromoquinolin-2-yl)ethanone (700 mg, 2.8 mmol),
dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine (230 mg, 0.560
mmol) and K.sub.2CO.sub.3 (1.16 g, 8.40 mmol) in dioxane (10 mL)
and water (2.500 mL) and the reaction mixture was refluxed at
110.degree. C. for 5 h. The reaction mixture was cooled to rt and
partitioned between sat.aq NH.sub.4Cl and EtOAc. The organic layer
was washed with NaHCO.sub.3 and brine and then dried (MgSO.sub.4),
filtered, concentrated and purified by flash silica gel
chromatography (loading solvent: DCM, eluted with 0.about.20%
EtOAc/hexanes) to yield 1-(6-(4-acetylphenyl)quinolin-2-yl)ethanone
(781 mg) as white solid. LC-MS retention time 3.776 min; m/z 290.29
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 8.35 (d,
J=8.3 Hz, 1H), 8.34 (d, J=8.3 Hz, 1H), 8.19 (d, J=8.5 Hz, 1H), 8.12
(d, J=8.3 Hz, 1H), 8.08-8.16 (m, 3H), 7.85 (d, J=8.3 Hz, 1H),
7.82-7.89 (m, 1H), 2.91 (s, 3H), 2.69 (s, 3H).
##STR00518##
Intermediate 161
2-Bromo-1-(6-(4-(2-bromoacetyl)phenyl)quinolin-2-yl)ethanone
[0816] Pyridinium tribromide (221 mg, 0.691 mmol) was added to a
suspension of 1-(6-(4-acetylphenyl)quinolin-2-yl)ethanone (100 mg,
0.346 mmol) and HBr (0.061 mL, 0.35 mmol) in acetic acid (3 mL) the
reaction mixture was stirred at 70.degree. C.) for 6 h. The
reaction was concentrated under vacuum and the residue was
partitioned between aq NaHCO.sub.3 (15 mL) and EtOAc (30 mL+10
mL+10 mL). The organic layers were combined, washed with brine,
dried, filtered and concentrated to yield crude
2-bromo-1-(6-(4-(2-bromoacetyl)phenyl)quinolin-2-yl)ethanone (120
mg) as white solid. LC-MS retention time 4.138 min; m/z 448.01
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00519##
Intermediate 162
(2S,4S)-2-(2-(6-(4-(2-((2S,4S)-1-(tert-Butoxycarbonyl)-4-methylpyrrolidine-
-2-carbonyloxy)acetyl)phenyl)quinolin-2-yl)-2-oxoethyl)1-tert-butyl
4-methylpyrrolidine-1,2-dicarboxylate
[0817] DIPEA (0.094 mL, 0.537 mmol) was added to a solution of
2-bromo-1-(6-(4-(2-bromoacetyl)phenyl)quinolin-2-yl)ethanone (60
mg, 0.134 mmol) and
(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid (64.6 mg, 0.282 mmol) in acetonitrile (2 mL) and the mixture
was stirred at rt for 16 h. The reaction mixture was partitioned
between EtOAc (20 mL+20 mL)/aq NaHCO.sub.3 (5 mL). The combined
organic layers were washed with aq NaHCO.sub.3, brine, dried,
filtered and concentrated to yield crude
(2S,4S)-2-(2-(6-(4-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrr-
olidine-2-carbonyloxy)acetyl)phenyl)quinolin-2-yl)-2-oxoethyl)1-tert-butyl
4-methylpyrrolidine-1,2-dicarboxylate (100 mg) as orange solid.
This material was used without further purification. LC-MS
retention time 4.558 min; m/z 742.36 (MH-). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time
of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode.
##STR00520##
Intermediate 163
(2S,4S)-tert-Butyl
2-(5-(4-(2-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1-
H-imidazol-4-yl)quinolin-6-yl)phenyl)-1H-imidazol-2-yl)-4-methylpyrrolidin-
e-1-carboxylate
[0818] Ammonium acetate (207 mg, 2.69 mmol) was added to a solution
of
(2S,4S)-2-(2-(6-(4-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-
e-2-carbonyloxy)acetyl)phenyl)quinolin-2-yl)-2-oxoethyl)1-tert-butyl
4-methylpyrrolidine-1,2-dicarboxylate (100 mg, 0.134 mmol) in
toluene (3 mL) and the reaction mixture was stirred at 120.degree.
C. (microwave reactor) for 3 h. The reaction was partitioned
between EtOAc (20 mL+10 mL+10 mL) and aq NaHCO.sub.3 (10 mL) and
the combined organic layers were dried, filtered and concentrated.
This crude product was purified by preparative HPLC
(H.sub.2O--CH.sub.3CN with 10 mM NH.sub.4OAc buffer) to yield
product (2S,4S)-tert-butyl
2-(5-(6-(4-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1-
H-imidazol-5-yl)phenyl)quinolin-2-yl)-1H-imidazol-2-yl)-4-methylpyrrolidin-
e-1-carboxylate (16.6 mg) as a yellow solid. LC-MS retention time
3.270 min; m/z 704.72 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00521##
Intermediate 164
2-(2-((2S,4S)-4-Methylpyrrolidin-2-yl)-1H-imidazol-4-yl)-6-(4-(2-((2S,4S)--
4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)quinoline
[0819] 4M HCl (0.2 mL, 0.800 mmol) in dioxane was added to a
solution of (2S,4S)-tert-butyl
2-(5-(6-(4-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1-
H-imidazol-5-yl)phenyl)quinolin-2-yl)-1H-imidazol-2-yl)-4-methylpyrrolidin-
e-1-carboxylate (16.6 mg, 0.024 mmol) in dioxane (1 mL) and the
reaction mixture was stirred at rt for 3 d. The reaction mixture
was concentrated to yield a crude HCl salt of
2-(2-((2S,4S)-4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)-6-(4-(2-((2S,4S)-
-4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)quinoline (19.1
mg) as yellow solid. LC-MS retention time 2.765 min; m/z 504.47
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00522##
##STR00523##
Intermediate 165
(2S,5S)-2-(2-(6-(4-(2-((2S,4S)-1-(tert-Butoxycarbonyl)-4-methylpyrrolidine-
-2-carbonyloxy)acetyl)phenyl)quinolin-2-yl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate
[0820] DIPEA (0.059 mL, 0.335 mmol) was added to a solution of
2-bromo-1-(6-(4-(2-bromoacetyl)phenyl)quinolin-2-yl)ethanone (50
mg, 0.11 mmol) and
(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxyli- c
acid (54 mg, 0.24 mmol) in acetonitrile (2 mL) and the reaction
mixture was stirred at rt for 16 h. The reaction was partitioned
between EtOAc (20+20 mL)/aq NaHCO.sub.3 (5 mL). The combined
organic layers were washed with sat. aq NaHCO.sub.3, brine, dried,
filtered and concentrated to yield crude
(2S,5S)-2-(2-(6-(4-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-
e-2-carbonyloxy)acetyl)phenyl)quinolin-2-yl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate (69 mg) as orange solid. This
crude was used without further purification. LC-MS retention time
4.483 min; m/z 744.61 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00524##
Intermediate 166
(2S,5S)-tert-Butyl
2-(5-(4-(2-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl)-1-
H-imidazol-4-yl)quinolin-6-yl)phenyl)-1H-imidazol-2-yl)-5-methylpyrrolidin-
e-1-carboxylate
[0821] Ammonium acetate (107 mg, 1.39 mmol) was added to a
suspension of
(2S,5S)-2-(2-(6-(4-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-
e-2-carbonyloxy)acetyl)phenyl)quinolin-2-yl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate (69 mg, 0.093 mmol) in
toluene (Volume: 2 mL) and the reaction mixture was stirred at
120.degree. C. (microwave reactor) for 3 h. The reaction was
partitioned between EtOAc (10 mL+5 mL+5 mL) and aq NaHCO3 (5 mL).
The combined organic layers were dried, filtered and concentrated
and the crude product was purified by preparative HPLC
(H.sub.2O--CH.sub.3CN with 10 mM NH.sub.4OAc buffer) to yield
product (2S,55)-tert-butyl
2-(4-(6-(4-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl)-1-
H-imidazol-4-yl)phenyl)quinolin-2-yl)-1H-imidazol-2-yl)-5-methylpyrrolidin-
e-1-carboxylate (13.4 mg) as a yellow solid. LC-MS retention time
4.350 min; m/z 702.29 (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold
time of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.37 (d, J=8.8 Hz, 1H),
8.16 (s, 1H), 8.07-8.10 (m, 2H), 8.03 (d, J=8.8 Hz, 1H), 7.79-7.88
(m, 5H), 7.40 (s, 1H), 4.90-5.07 (m, 2H), 3.99-4.13 (m, 2H),
2.10-2.39 (m, 6H), 1.68-1.82 (m, 2H), 1.43 (d, J=6.0 Hz, 6H),
1.24-1.59 (m, 22H).
##STR00525##
Intermediate 167
2-(2-((2S,5S)-5-Methylpyrrolidin-2-yl)-1H-imidazol-4-yl)-6-(4-(2-((2S,5S)--
5-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)quinoline
[0822] 4M HCl (0.2 mL, 0.8 mmol) in dioxane was added to a solution
of (2S,5S)-tert-butyl
2-(5-(6-(4-(2-((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl)-1-
H-imidazol-5-yl)phenyl)quinolin-2-yl)-1H-imidazol-2-yl)-5-methylpyrrolidin-
e-1-carboxylate (12.7 mg, 0.018 mmol) in dioxane (1 mL) and the
mixture was stirred at rt for 3 d. The reaction mixture was
concentrated to yield a crude HCl salt of
2-(2-((2S,5S)-5-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)-6-(4-(2-((2S,5S)-
-5-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)quinoline (18.1
mg) as yellow solid. LC-MS retention time 2.597 min; m/z 504.46
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.09 (d, J=9.0 Hz,
1H), 8.66 (s, 1H), 8.54-8.61 (m, 2H), 8.51 (d, J=9.0 Hz, 1H), 8.50
(d, J=8.8 Hz, 1H), 8.00-8.08 (m, 4H), 7.99 (s, 1H), 5.14 (t, J=8.5
Hz, 1H), 5.07 (t, J=8.0 Hz, 1H), 3.90-4.04 (m, 2H), 2.52-2.75 (m,
4H), 2.39-2.53 (m, 2H), 1.98-2.14 (m, 2H), 1.59 (dd, J=9.7, 6.7 Hz,
6H).
##STR00526##
[0823] Intermediate 173 was prepared in an analogous manner to
Intermediate 139 in Scheme 35 by utilizing
6-bromo-2-chloroquinoxaline as the starting material rather than
6-bromo-2-chloroquinoline. Analytical data shown below.
##STR00527##
Intermediate 168
6-Bromo-2-(1-ethoxyvinyl)quinoxaline
[0824] The reaction yielded the desired product (380 mg) as a white
solid. LC-MS retention time 4.178 min; m/z 278.9, 280.98 (1:1)
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00528##
Intermediate 169
2-Bromo-1-(6-bromoquinoxalin-2-yl)ethanone
[0825] The reaction yielded the desired product (390 mg) as a white
solid. LC-MS retention time 3.743 min; m/z 331.12 (1:2:1) (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00529##
Intermediate 170
(1R,3S,5R)-3-(2-(6-Bromoquinoxalin-2-yl)-2-oxoethyl)2-tert-butyl
2-azabicyclo[3.1.0]hexane-2,3-dicarboxylate
[0826] The reaction yielded the desired product (270 mg) as a
pink/orange solid. LC-MS retention time 4.115 min; m/z 473, 475.81
(1:1) (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold
time of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00530##
Intermediate 171
tert-Butyl
(1R,3S,5R)-3-(4-(6-bromo-2-quinoxalinyl)-1H-imidazol-2-yl)-2-az-
abicyclo[3.1.0]hexane-2-carboxylate
[0827] The reaction yielded the desired product (125 mg) as a
red/orange solid. LC-MS retention time 3.958 min; m/z 453.90,
455.87 (1:1) (MH-). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold
time of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.40 (br. s., 1H), 8.22 (d,
J=2.0 Hz, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.93-7.99 (m, 1H), 7.90 (dd,
J=8.8, 2.0 Hz, 1H), 4.71-4.81 (m, 1H), 3.59 (br. s., 1H), 2.49-2.63
(m, 1H), 2.31-2.45 (m, 1H), 1.68-1.82 (m, 1H), 1.16-1.53 (m, 5H),
0.81-0.94 (m, 1H), 0.64 (br. s., 1H).
##STR00531##
Intermediate 172
tert-Butyl
(1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-2--
azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)-2-quinoxalinyl)-1H-imi-
dazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0828] The reaction yielded the desired product (64.4 mg) as a
bright yellow solid. LC-MS retention time 3.696 min; m/z 699.3
(MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold
time of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.45 (s, 1H), 8.31
(s, 1H), 8.19 (d, J=8.3 Hz, 1H), 8.07 (d, J=8.8 Hz, 1H), 7.97 (s,
1H), 7.81-7.95 (m, 4H), 7.60 (s, 1H), 4.57-4.80 (m, 2H), 3.23-3.60
(m, 2H), 2.23-2.48 (m, 4H), 1.61-1.75 (m, 2H), 1.28 (br. s., 18H),
0.73-0.87 (m, 2H), 0.59 (br. s., 2H).
##STR00532##
Intermediate 173
Methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)pheny-
l)-2-quinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-met-
hylpropyl)carbamate
[0829] The reaction yielded an HCl salt of the desired product (80
mg) as a yellow solid. LC-MS retention time 2.915 min; m/z 501.42
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.54 (s, 1H),
8.40-8.43 (m, 1H), 8.34 (s, 1H), 8.23-8.30 (m, 2H), 8.13 (s, 1H),
8.00-8.08 (m, 4H), 4.88-5.08 (m, 2H), 3.53-3.77 (m, 2H), 2.64-2.90
(m, 4H), 2.08-2.22 (m, 2H), 1.23-1.36 (m, 2H), 1.02-1.14 (m,
2H).
##STR00533##
##STR00534##
Intermediate 174
(1R,3S,5R)-tert-Butyl
3-(4-((6-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1H--
imidazol-4-yl)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate
[0830] Compound was prepared in an analogous manner to Intermediate
153 with the appropriate starting materials to yield a TFA salt of
the title compound (121 mg) as a yellow glass. LC-MS retention time
4.100 min; m/z 647.37 (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00535##
Intermediate 175
(1R,3S,5R)-3-(4-((6-(2-((2S,4S)-4-Methylpyrrolidin-2-yl)-1H-imidazol-4-yl)-
naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
[0831] 2M HCl (0.33 mL, 0.66 mmol) in dioxane was added to a
solution of a TFA salt of (1R,3S,5R)-tert-butyl
3-(5-((6-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1H--
imidazol-4-yl)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hexane-2-carboxylate (115 mg, 0.131 mmol) in dioxane (2 mL) and
the reaction was stirred vigorously overnight. The reaction was
concentrated to yield an HCl salt of
(1R,3S,5R)-3-(4-((6-(2-((2S,4S)-4-methylpyrrolidin-2-yl)-1H-imidazol-4-yl-
)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(87 mg) which was used without further purification. LC-MS
retention time 2.793 min; m/z 449.41 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00536##
[0832] Intermediate 178 was prepared in an analogous manner to the
preparation of Intermediate 117 in Scheme 29 utilizing
(S)-2-(tert-butoxycarbonylamino)-3-methylbutanoic acid as a
starting material rather than
(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic
acid. Analytical data for the intermediates shown below.
##STR00537##
Intermediate 176
(S)-2-(4-(6-(2-((S)-2-(tert-Butoxycarbonylamino)-3-methylbutanoyloxy)acety-
l)naphthalen-2-yl)phenyl)-2-oxoethyl
2-(tert-butoxycarbonylamino)-3-methylbutanoate
[0833] The reaction yielded the desired product (700 mg) as a tan
solid. LC-MS retention time 2.243 min; m/z 717.5 (MH-). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 1.0 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00538##
Intermediate 177
tert-Butyl((1S)-1-(4-(4-(6-(2-((1S)-1-((tert-butoxycarbonyl)amino)-2-methy-
lpropyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-methylpr-
opyl)carbamate
[0834] The reaction yielded the desired product (453 mg) as a
yellow solid. LC-MS retention time 2.407 min; m/z 677.6 (MH-). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 1.0 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 5% MeOH/95% H.sub.2O/10
mM ammonium acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00539##
Intermediate 178
(S)-1-(5-(4-(6-(2-((S)-1-Amino-2-methylpropyl)-1H-imidazol-4-yl)naphthalen-
-2-yl)phenyl)-1H-imidazol-2-yl)-2-methylpropan-1-amine
[0835] The reaction yielded an HCl salt of the desired product (390
mg) as a yellow solid. LC-MS retention time 2.080 min; m/z 477.4
(MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1.0 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold
time of 1 min, and an analysis time of 3 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00540##
[0836] Intermediate 181 was prepared in an analogous manner to the
preparation of Intermediate 117 in Scheme 29 utilizing
(S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid as a
starting material rather than
(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic
acid. Analytical data for the intermediates shown below.
##STR00541##
Intermediate 179
(S)-3-(2-(4-(6-(2-((S)-4-(tert-Butoxycarbonyl)morpholine-3-carbonyloxy)ace-
tyl)naphthalen-2-yl)phenyl)-2-oxoethyl)4-tert-butyl
morpholine-3,4-dicarboxylate
[0837] The reaction yielded an HCl salt of the desired product
(1.60 g) as an off-white solid foam. LC-MS retention time 4.413
min; m/z 770.29 (MNa+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 8.47 (s, 1H),
8.13 (s, 1H), 7.98-8.11 (m, 5H), 7.86 (d, J=8.3 Hz, 3H), 5.39-5.70
(m, 4H), 4.66-4.83 (m, 2H), 4.46-4.63 (m, 2H), 3.91-4.03 (m, 2H),
3.68-3.85 (m, 4H), 3.49-3.59 (m, 3H), 3.27-3.40 (m, 1H), 1.51 (s,
18H).
##STR00542##
Intermediate 180
(R)-tert-Butyl
3-(5-(4-(6-(2-((R)-4-(tert-butoxycarbonyl)morpholin-3-yl)-1H-imidazol-4-y-
l)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)morpholine-4-carboxylate
[0838] The reaction yielded the desired product (1.12 g) as an
orange solid. LC-MS retention time 3.608 min; m/z 707.94 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.22 (br. s., 1H), 8.09 (s,
1H), 7.90-7.97 (m, 2H), 7.75-7.89 (m, 6H), 7.49 (br. s., 1H), 7.41
(br. s., 1H), 5.17-5.24 (m, 2H), 4.42 (t, J=11.2 Hz, 2H), 3.82-3.95
(m, 6H), 3.59 (tt, J=11.6, 3.0 Hz, 2H), 3.39-3.51 (m, 2H), 1.50 (d,
J=2.8 Hz, 18H).
##STR00543##
Intermediate 181
(R)-3-(5-(4-(6-(2-((R)-Morpholin-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)ph-
enyl)-1H-imidazol-2-yl)morpholine
[0839] The reaction yielded an HCl salt of the desired product (312
mg) as a yellow solid. LC-MS retention time 2.988 min; m/z 507.72
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00544##
[0840] Intermediate 184 was prepared in an analogous manner to the
preparation of Intermediate 117 in Scheme 29 utilizing
(S)-1-(tert-butoxycarbonyl)piperidine-2-carboxylic acid as a
starting material rather than
(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic
acid. Analytical data for the intermediates shown below.
##STR00545##
Intermediate 182
(S)-2-(2-(4-(6-(2-((S)-1-(tert-Butoxycarbonyl)piperidine-2-carbonyloxy)ace-
tyl)naphthalen-2-yl)phenyl)-2-oxoethyl)1-tert-butyl
piperidine-1,2-dicarboxylate
[0841] The reaction yielded the desired product (1.67 g) as a
solidified light yellow foam. LC-MS retention time 4.850 min; m/z
766.37 (MNa+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, chloroform-d) .delta. ppm 8.47 (s, 1H),
8.13 (s, 1H), 7.99-8.10 (m, 5H), 7.83-7.89 (m, 3H), 5.32-5.67 (m,
4H), 4.92-5.13 (m, 2H), 3.93-4.11 (m, 2H), 3.00-3.28 (m, 2H), 2.40
(br. s., 2H), 1.68-1.84 (m, 6H), 1.49 (s, 18H), 1.39-1.67 (m,
4H)
##STR00546##
Intermediate 183
(S)-tert-Butyl
2-(5-(4-(6-(2-((S)-1-(tert-butoxycarbonyl)piperidin-2-yl)-1H-imidazol-4-y-
l)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)piperidine-1-carboxylate
[0842] The reaction yielded the desired product (1.18 g) as a
bright yellow solid. LC-MS retention time 3.778 min; m/z 703.97
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.22 (br. s., 1H),
8.09 (s, 1H), 7.93 (t, J=7.7 Hz, 2H), 7.73-7.88 (m, 6H), 7.46 (br.
s., 1H), 7.38 (br. s., 1H), 5.47 (br. s., 2H), 4.03-4.15 (m, 2H),
2.99-3.12 (m, 2H), 2.46 (br. s., 2H), 1.80-1.94 (m, 2H), 1.50 (d,
J=2.3 Hz, 18H), 1.44-1.76 (m, 8H).
##STR00547##
Intermediate 184
(S)-2-(5-(4-(6-(2-((S)-Piperidin-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)ph-
enyl)-1H-imidazol-2-yl)piperidine
[0843] The reaction yielded an HCl salt of the desired product (280
mg) as a yellow solid. LC-MS retention time 3.015 min; m/z 503.69
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.06 (br.
s., 2H), 9.85 (br. s., 2H), 8.48 (s, 1H), 8.36 (s, 1H), 8.09-8.19
(m, 3H), 7.96-8.09 (m, 7H), 4.70 (br. s., 2H), 3.42-3.53 (m, 2H),
3.14 (br. s., 2H), 2.10-2.37 (m, 4H), 1.60-2.00 (m, 8H).
##STR00548##
[0844] Intermediate 187 was prepared in an analogous manner to the
preparation of Intermediate 117 in Scheme 29 utilizing
(2S,5R)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic
acid (prepared according to JOC 1995, p. 5011) as a starting
material rather than
(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic
acid. Analytical data for the intermediates shown below.
##STR00549##
Intermediate 185
(2S,5R)-2-(2-(4-(6-(2-((2S,5R)-1-(tert-Butoxycarbonyl)-5-methylpyrrolidine-
-2-carbonyloxy)acetyl)naphthalen-2-yl)phenyl)-2-oxoethyl)1-tert-butyl
5-methylpyrrolidine-1,2-dicarboxylate
[0845] The reaction yielded the desired product (979 mg). LC-MS
retention time 4.796 min; m/z 766.19 (MNa+). LC data was recorded
on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, chloroform-d) .delta. ppm
8.47 (br. s., 1H), 8.13 (d, J=2.5 Hz, 1H), 7.99-8.11 (m, 5H), 7.86
(dd, J=7.8, 2.3 Hz, 3H), 5.21-5.77 (m, 4H), 4.51-4.58 (m, 1H), 4.47
(dd, J=7.8, 4.5 Hz, 1H), 4.17-4.26 (m, 1H), 4.07-4.14 (m, 1H),
2.23-2.48 (m, 6H), 1.60-1.68 (m, 2H), 1.44-1.52 (m, 18H), 1.25 (d,
J=6.3 Hz, 3H), 1.21 (d, J=6.3 Hz, 3H).
##STR00550##
Intermediate 186
(2S,5R)-tert-Butyl
2-(5-(4-(6-(2-((2S,5R)-1-(tert-butoxycarbonyl)-5-methylpyrrolidin-2-yl)-1-
H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-5-methylpyrrolid-
ine-1-carboxylate
[0846] The reaction yielded the desired product (561 mg) as an
orange solid foam. LC-MS retention time 3.673 min; m/z 703.95
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00551##
Intermediate 187
2-((2S,5R)-5-Methylpyrrolidin-2-yl)-5-(4-(6-(2-((2S,5R)-5-methylpyrrolidin-
-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazole
[0847] The reaction yielded an HCl salt of the desired product (157
mg) as a light orange solid. LC-MS retention time 2.975 min; m/z
503.71 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 9.84-10.25
(m, 4H), 8.54 (s, 1H), 8.36 (s, 1H), 7.95-8.22 (m, 10H), 5.15 (br.
s., 2H), 4.00 (br. s., 2H), 2.53-2.63 (m, 4H), 2.30-2.41 (m, 2H),
1.67-1.82 (m, 2H), 1.43 (d, J=4.3 Hz, 6H).
##STR00552##
[0848] Intermediate 189 was prepared as shown in Scheme 49
utilizing
(S)-1-(tert-butoxycarbonyl)-4-methylenepyrrolidine-2-carboxylic
acid as starting material. Analytical data for the intermediates
shown below.
##STR00553##
Intermediate 188
tert-Butyl
(2S)-2-(4-(4-(6-(2-((2S)-1-(tert-butoxycarbonyl)-4-methylene-2--
pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-met-
hylene-1-pyrrolidinecarboxylate
[0849] Hunig's base (0.43 mL, 2.47 mmol) was added to a stirred
solution of
(S)-1-(tert-butoxycarbonyl)-4-methylenepyrrolidine-2-carboxylic
acid (509 mg, 2.241 mmol) and
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (500
mg, 1.121 mmol) in MeCN (25 mL). The mixture was stirred for 18 h
at RT. The solvent was removed in vacuo and the residue was taken
up in ethyl acetate and washed with water, saturated sodium
bicarbonate solution, and brine. After being concentrated, the
residue was taken up in xylene (25 mL) and ammonium acetate (1.3 g,
16.85 mmol) was added.
[0850] The pressure vessel was sealed and heated at 140.degree. C.
for 2.5 h. The reaction mixture was taken up in ethyl acetate and
washed with saturated sodium bicarbonate solution and brine. After
being concentrated, the crude product was charged (methylene
chloride) to a 80 g Thompson silica gel cartridge (eluted with 15%
B to 100% B over 1 L where Solvent B=ethyl acetate and Solvent
A=hexanes) to yield tert-butyl
(2S)-2-(4-(4-(6-(2-(2S)-1-(tert-butoxycarbonyl)-4-methylene-2-pyrrolidiny-
l)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methylene-1-py-
rrolidinecarboxylate (380 mg, 45% yield). LC-MS retention time 3.32
min; calcd. for C.sub.42H.sub.47N.sub.6O.sub.4: 699.36 m/z Found
699.41 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
methanol/95% water/0.1% TFA and Solvent B was 95% methanol/5%
water/0.1% TFA. MS data was determined using a
MICROMASS.RTM.Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.21 (s, 1H), 8.14 (s, 1H), 7.98 (s,
1H), 7.97 (s, 1H), 7.87-7.83 (m, 6H), 7.54 (s, 1H), 7.46 (s, 1H),
5.14 (s, 2H), 5.19 (s, 2H), 4.30 (br s, 2H), 4.15 (br s, 2H),
3.22-3.16 (m, 2H), 2.81-2.76 (m, 2H), 1.51/1.31 (s, 18H).
##STR00554##
Intermediate 189
2-((S)-4-Methylenepyrrolidin-2-yl)-4-(4-(6-(2-((S)-4-methylenepyrrolidin-2-
-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazole
[0851] A solution of 4N HCl in dioxane (10 mL) was added to
tert-butyl
(2S)-2-(4-(4-(6-(2-((2S)-1-(tert-butoxycarbonyl)-4-methylene-2-pyrrolidin-
yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methylene-1-p-
yrrolidinecarboxylate (380 mg. 0.544 mmol) in MeOH (10 mL) and
stirred at ambient conditions for 4 hours, concentrated, and dried
under vacuum. Tetra HCl salt (assume theoretical: 350 mg). LC-MS
retention time 2.68 min; calcd. for C.sub.32H.sub.31N.sub.6: 499.26
m/z Found 499.21 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
methanol/95% water/0.1% TFA and Solvent B was 95% methanol/5%
water/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00555##
[0852] Intermediate 191 was prepared as shown in Scheme 50
utilizing (S)-2-(tert-butoxycarbonylamino)propanoic acid as
starting material. Analytical data for the intermediates shown
below.
##STR00556##
Intermediate 190
tert-Butyl((1S)-1-(4-(4-(6-(2-((1S)-1-((tert-butoxycarbonyl)amino)ethyl)-1-
H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)carbamate
[0853] Hunig's base (0.39 mL, 2.24 mmol) was added to a stirred
solution of (S)-2-(tert-butoxycarbonylamino)propanoic acid (424 mg,
2.24 mmol) and
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (500
mg, 1.121 mmol) in MeCN (50 mL). The mixture was stirred for 18 h
at RT. The solvent was removed in vacuo and the residue was taken
up in ethyl acetate and washed with water, saturated sodium
bicarbonate solution, and brine. After being concentrated, the
residue was taken up in xylene (25 mL) and ammonium acetate (1.5 g,
20.18 mmol) was added. The pressure vessel was sealed and heated at
140.degree. C. for 3.5 h. The reaction mixture was taken up in
ethyl acetate and washed with saturated sodium bicarbonate solution
and brine. After being concentrated, the crude product was charged
(methylene chloride) to a 90 g
[0854] Thompson silica gel cartridge (eluted with 15% B to 100% B
over 1 L where Solvent B=ethyl acetate and Solvent A=hexanes) to
provide
tert-Butyl((1S)-1-(4-(4-(6-(2-((1S)-1-((tert-butoxycarbonyl)amino)ethyl)--
1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)carbamate
(368 mg, 47% yield). NOTE: Sample was only partially soluble in
methylene chloride. LC-MS retention time 3.24 min; calcd. for
C.sub.36H.sub.43N.sub.6O.sub.4: 623.33 m/z Found 623.37
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.20 (s,
1H), 8.11 (s, 1H), 7.95 (t, J=8.24 Hz, 2H), 7.86-7.79 (m, 6H), 7.46
(s, 1H), 7.37 (s, 1H), 4.93-4.89 (m, 2H), 1.57-1.48 (m, 24H).
##STR00557##
Intermediate 191
(S)-1-(4-(4-(6-(2-((S)-1-Aminoethyl)-1H-imidazol-4-yl)naphthalen-2-yl)phen-
yl)-1H-imidazol-2-yl)ethanamine
[0855] A solution of 4N HCl in dioxane (10 mL) was added to
tert-butyl((1S)-1-(4-(4-(6-(2-((1S)-1-((tert-butoxycarbonyl)amino)ethyl)--
1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)carbamate
(368 mg. 0.59 mmol) in MeOH (10 mL) and stirred at ambient
conditions for 5 hours, concentrated, and dried under vacuum. Tetra
HCl salt (assume theoretical: 334 mg). LC-MS retention time 1.33
min; calcd. for C.sub.26H.sub.27N.sub.6: 423.23 m/z Found 423.17
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 2.0.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 1 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min and an
analysis time of 3 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00558##
[0856] Intermediate 193 was prepared as shown in Scheme 51
utilizing (S)-2-(tert-butoxycarbonyl(methyl)amino)propanoic acid as
starting material. Analytical data for the intermediates shown
below.
##STR00559##
Intermediate 192
tert-Butyl((1S)-1-(4-(4-(6-(2-((1S)-1-((tert-butoxycarbonyl)(methyl)amino)-
ethyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)methylc-
arbamate
[0857] Hunig's base (0.78 mL, 4.48 mmol) was added to a stirred
solution of (S)-2-(tert-butoxycarbonyl(methyl)amino)propanoic acid
(456 mg, 2.241 mmol) and
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (500
mg, 1.121 mmol) in MeCN (50 mL). The mixture was stirred for 18 h
at RT. The solvent was removed in vacuo and the residue was taken
up in ethyl acetate and washed with water, saturated sodium
bicarbonate solution, and brine. After being concentrated, the
residue was taken up in xylene (25 mL) and ammonium acetate (859
mg, 11.15 mmol) was added. The pressure vessel was sealed and
heated at 140.degree. C. for 3 h. The reaction mixture was taken up
in ethyl acetate and washed with saturated sodium bicarbonate
solution and brine. After being concentrated, the crude product was
charged (methylene chloride) to a 90 g Thompson silica gel
cartridge (eluted with 15% B to 100% B over 1 L where Solvent
B=ethyl acetate and Solvent A=hexanes) to yield
tert-butyl((1S)-1-(4-(4-(6-(2-((1S)-1-((tert-butoxycarbonyl)(methyl)amino-
)ethyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)methyl-
carbamate (460 mg, 58% yield). LC-MS retention time 3.28 min;
calcd. for C.sub.38H.sub.47N.sub.6O.sub.4: 651.37 m/z Found 651.34
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm
8.24-8.12 (m, 2H), 7.98-7.82 (m, 8H), 7.52-7.42 (m, 2H), 5.47 (br
s, 2H), 2.77 (s, 3H), 2.76 (s, 3H), 1.64 (m, 6H), 1.52 (s,
18H).
##STR00560##
Intermediate 193
(S)--N-Methyl-1-(4-(4-(6-(2-((S)-1-(methylamino)ethyl)-1H-imidazol-4-yl)na-
phthalen-2-yl)phenyl)-1H-imidazol-2-yl)ethanamine
[0858] A solution of 4N HCl in dioxane (10 mL) was added to
tert-butyl((1S)-1-(4-(4-(6-(2-((1S)-1-((tert-butoxycarbonyl)(methyl)amino-
)ethyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)methyl-
carbamate (434 mg. 0.667 mmol) in MeOH (10 mL) and stirred at
ambient conditions for 3 hours, concentrated and dried under
vacuum. Tetra HCl salt (assume theoretical: 396 mg). LC-MS
retention time 1.44 min; calcd. for C.sub.28H.sub.31N.sub.6: 451.26
m/z Found 451.17 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min and an analysis time of 3 min where Solvent A was 5%
methanol/95% water/0.1% TFA and Solvent B was 95% methanol/5%
water/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00561##
[0859] Intermediate 196 was prepared as shown in Scheme 52
utilizing
(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid as starting material. Analytical data for the intermediates
shown below.
##STR00562##
Intermediate 194
(2S,4S)-1-(tert-Butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid
[0860] A solution of
(S)-1-(tert-butoxycarbonyl)-4-methylenepyrrolidine-2-carboxylic
acid (4 g, 17.60 mmol) in 2-propanol (10 mL) was added to a
nitrogen purged suspension of 10% palladium on carbon (936 mg) in
2-propanol (240 mL) and the flask was charged with hydrogen gas (1
atm). After being stirred 18, the catalyst was removed by
filtration over CELITE.RTM. and the filtrate concentrated. LC
analysis showed the sample contained .about.14% of the
trans-isomer, and recrystallization from toluene enriched the
cis-isomer to 96% (16:1). LC-MS retention time 3.26 min; calcd. for
C.sub.11H.sub.20N.sub.6O.sub.4: 230.14 m/z Found 252.14
[M+Na].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. 2.times.5 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min and an
analysis time of 5 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 90% methanol/10% water/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm
4.21-4.17 (m, 1H), 3.76-3.67 (m, 1H), 2.96-2.92 (m, 1H), 2.49-2.46
(m, 1H), 2.30-2.29 (m, 1H), 1.59-1.51 (m, 1H), 1.47/1.43 (m, 9H),
1.10-1.06 (m, 3H).
##STR00563##
Intermediate 195
tert-Butyl
(2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methyl-
-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4--
methyl-1-pyrrolidinecarboxylate
[0861] Hunig's base (0.381 mL, 2.181 mmol) was added to a stirred
solution of
(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid (500 mg, 2.18 mmol) and
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (486
mg, 1.09 mmol) in acetonitrile (11 mL). The heterogeneous mixture
was stirred for 18 h at RT and additional Hunig's base (1 equiv)
was added and the mixture was stirred for 24 h. The solvent was
removed in vacuo and the residue was taken up in methylene chloride
and washed with saturated sodium bicarbonate solution, brine, and
dried over sodium sulfate. Wash was repeated 2.times.. The crude
product, isolated as a tan foam, was taken up in xylene (15 mL) and
ammonia acetate (1.261 g, 16.36 mmol) was added. The pressure
vessel was sealed and placed into a preheated oil bath (140.degree.
C.) and stirred for 2 h. The reaction mixture was taken up in ethyl
acetate and washed with saturated sodium bicarbonate solution and
brine. After being concentrated, the crude product was charged
(methylene chloride) to a 80 g Thompson silica gel cartridge
(eluted with 25% B to 100% B over 1.5 L and hold 0.5 L B. Solvent
B=ethyl acetate and Solvent A=hexanes) to yield tert-butyl
(2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-p-
yrrolidinecarboxylate
[0862] (214.2 mg 27.9%) as a yellow foam. LC-MS retention time 3.32
min; calcd. for C.sub.42H.sub.51N.sub.6O.sub.4: 703.40 m/z Found
703.28 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
methanol/95% water/0.1% TFA and Solvent B was 95% methanol/5%
water/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.20 (s, 1H), 8.12 (s, 1H), 7.96-7.94 (m, 2H),
7.87-7.82 (m, 6H), 7.50 (s, 1H), 7.42 (s, 1H), 4.87 (br. s, 2H),
3.81 (br. s, 2H), 3.24-3.18 (m, 2H), 2.53-2.50 (m, 2H), 2.36 (br s,
2H), 1.82-1.74 (m, 2H), 1.46/1.23 (s, 18H), 1.17-1.16 (m, 6H).
##STR00564##
Intermediate 196
2-((2S,4S)-4-Methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
[0863] A solution of 4N HCl in dioxane (15 mL) was added to
tert-butyl
(2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-p-
yrrolidinecarboxylate (200 mg. 0.285 mmol) in MeOH (5 mL) and
stirred at ambient conditions for 2 hours, concentrated, and dried
under vacuum. Tetra HCl salt (assume theoretical: 185 mg). LC-MS
retention time 2.64 min; calcd. for C.sub.32H.sub.35N.sub.6: 503.29
m/z Found 503.20 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
methanol/95% water/0.1% TFA and Solvent B was 95% methanol/5%
water/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00565##
[0864] Intermediate 201 was prepared as shown in Scheme 53
utilizing
(2S,4R)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid as starting material. Analytical data for the intermediates
shown below.
##STR00566##
Intermediate 197
[0865] The trans-isomer was isolated from a mixture of cis, trans,
and the dimethyl analog which could be obtained from methyl iodide
alkylation of (S)-1-tert-butyl
2-methyl-5-oxopyrrolidine-1,2-dicarboxylate according to the
procedure described in Tetrahedron Letters, 2003, 3203-3205.
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 4.66-4.64 (m, 1H), 3.80 (s,
3H), 2.34-2.29 (m, 1H), 2.04-1.99 (m, 1H), 1.49 (s, 9H), 1.21-1.19
(m, 3H).
##STR00567##
Intermediate 198
[0866] Borane-methyl sulfide complex (17.67 mL, 35.3 mmol) was
added to a solution of Intermediate 197 (6.06 g, 23.55 mmol) in THF
(180 mL), and the reaction mixture was heated at 40.degree. C. for
16 hr. The solvent was removed in vacuo and the residue was
partitioned between EtOAc and water (250 mL each). The aqueous
layer was extracted with EtOAc (2.times.60 mL), and the combined
organic phase was dried with Na.sub.2SO.sub.4, and concentrated in
vacuo. The resultant colorless oil was purified with a flash
chromatography (10-65% EtOAc/Hexane) to afford Intermediate 198 as
a colorless oil (3.65 g). .sup.1H NMR (CDCl.sub.3, .delta.=7.24
ppm, 400 MHz): 4.36-4.33 (dd, J=2.4, 10, 0.4H), 4.26-4.23 (dd, J=3,
8.9, 0.6H), 3.73-3.63 ('s' overlapped with `m`, 4H), 2.97-2.85 (m,
1H), 2.37 (m, 1H), 2.04 (m, 1H), 1.81 (m, 1H), 1.44-1.39 (two `s`,
9H), 1.03-1.00 (two `s`, 3H).
##STR00568##
Intermediate 199
(2S,4R)-1-(tert-Butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid
[0867] To a solution of ester Intermediate 198 (3.63 g, 14.92 mmol)
in ethanol (36 mL) was added solution of LiOH (0.393 g, 10.42 mmol)
in water (18.00 mL), and the reaction mixture was stirred at room
temperature for 22 hr. The organic solvent was evaporated in vacuo
and the residue was diluted with water (30 mL) and washed with
ethylacetate (50 mL). It was chilled in ice-water bath, and
acidified to a pH range of .about.2 with 1N HCl. It was then
extracted with EtOAc (50 mL, 2.times.). The organic layer was dried
with Na.sub.2SO.sub.4 and concentrated in vacuo to afford
Intermediate 199 as a colorless oil, which became a white solid
upon extended exposure to high vacuum (3.3 g). .sup.1H NMR
(CDCl.sub.3, .delta.=7.24 ppm, 400 MHz): 4.37-4.27 (overlapped `m`
1H), 3.72-3.50 (m, 1H), 3.01-2.83 (overlapped `m`, 1H), 2.50-2.11
(m, 2H), 1.89-1.57 (m, 1H), 1.47-1.39 (two `s`, 9H), 1.04-1.03 (two
`s`, 3H).
##STR00569##
Intermediate 200
tert-Butyl
(2S,4R)-2-(4-(4-(6-(2-((2S,4R)-1-(tert-butoxycarbonyl)-4-methyl-
-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4--
methyl-1-pyrrolidinecarboxylate
[0868] Hunig's base (0.381 mL, 2.181 mmol) was added to a stirred
solution of
(2S,4R)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid (500 mg, 2.18 mmol) and
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (486
mg, 1.09 mmol) in acetonitrile (11 mL). The heterogeneous mixture
was stirred for 18 h at RT and additional Hunig's base (1 eqv) was
added and the mixture was stirred for 24 h. The solvent was removed
in vacuo and the residue was taken up in methylene chloride and
washed with saturated sodium bicarbonate solution, brine, and dried
over sodium sulfate. Wash was repeated 2.times.. The crude product,
isolated as a tan foam, was taken up in xylene (15 mL) and ammonia
acetate (1.261 g, 16.36 mmol) was added. The pressure vessel was
sealed and placed into a preheated oil bath (140.degree. C.) and
stirred for 2 h. The reaction mixture was taken up in ethyl acetate
and washed with saturated sodium bicarbonate solution and brine.
After being concentrated, the crude product was charged (methylene
chloride) to a 80 g Thompson silica gel cartridge (eluted with 25%
B to 100% B over 1.5 L and hold 0.5 L B. Solvent B=ethyl acetate
and Solvent A=hexanes) to yield tert-butyl
(2S,4R)-2-(4-(4-(6-(2-((2S,4R)-1-(tert-butoxycarbonyl)-4-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-p-
yrrolidinecarboxylate (214.2 mg 27.9%) as a yellow foam. LC-MS
retention time 3.32 min; calcd. for C.sub.42H.sub.51N.sub.6O.sub.4:
703.40 m/z Found 703.28 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 2.0.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min and an analysis time of 5 min
where Solvent A was 5% methanol/95% water/0.1% TFA and Solvent B
was 95% methanol/5% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.20 (s, 1H), 8.12 (s, 1H), 7.96-7.94
(m, 2H), 7.87-7.82 (m, 6H), 7.50 (s, 1H), 7.42 (s, 1H), 4.87 (br.
s, 2H), 3.81 (br. s, 2H), 3.24-3.18 (m, 2H), 2.53-2.50 (m, 2H),
2.36 (br s, 2H), 1.82-1.74 (m, 2H), 1.46/1.23 (s, 18H), 1.17-1.16
(m, 6H).
##STR00570##
Intermediate 201
2-((2S,4R)-4-Methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4R)-4-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
[0869] A solution of 4N HCl in dioxane (15 mL) was added to
tert-butyl
(2S,4R)-2-(4-(4-(6-(2-((2S,4R)-1-(tert-butoxycarbonyl)-4-methyl-2-pyrroli-
dinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-p-
yrrolidinecarboxylate (200 mg. 0.285 mmol) in MeOH (5 mL) and
stirred at ambient conditions for 2 hours, concentrated, and dried
under vacuum. Tetra HCl salt (assume theoretical: 185 mg). LC-MS
retention time 2.64 min; calcd. for C.sub.32H.sub.35N.sub.6: 503.29
m/z Found 503.20 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
methanol/95% water/0.1% TFA and Solvent B was 95% methanol/5%
water/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00571##
Intermediate 203 was prepared as shown in Scheme 54 utilizing as
starting material
(1S,3S,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-c-
arboxylic acid (which could be prepared according to WO
2004/052850). Analytical data for the intermediates shown
below.
##STR00572##
[0870] Intermediate 202
tert-Butyl
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-(tert-butoxycarbonyl)-2--
azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazo-
l-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0871] Hunig's base (0.384 mL, 2.2 mmol) was added to a stirred
solution of
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone
(0.491 g, 1.100 mmol) and
(1S,3S,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic
acid (500 mg, 1.1 mmol) in acetonitrile (11 mL). The heterogeneous
mixture was stirred for 20 h at RT the solvent was removed in
vacuo. The residue was taken up in ethyl acetate and washed with
saturated sodium bicarbonate solution, brine, and dried over sodium
sulfate. Wash was repeated 2.times.. The crude product was taken up
in xylene (11 mL) and ammonia acetate (1.27 g, 16.5 mmol) was
added. The pressure vessel was sealed and placed into a preheated
oil bath (140.degree. C.) and stirred for 2 h. The solvent was
removed in vacuo and the residue was taken up in ethyl acetate and
washed with saturated sodium bicarbonate solution and brine. After
being concentrated, the crude product was charged (methylene
chloride) to a 80 g Thompson silica gel cartridge (eluted with 25%
B to 100% B over 1.5 L and hold 0.5 L B. Solvent B=ethyl acetate
and Solvent A=hexanes) to yield tert-butyl
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2--
azabicyclo[3.1.0]hexane-2-carboxylate (418.9 mg, 43%) as a light
yellow solid. LC-MS retention time 3.46 min; calcd. for
C.sub.42H.sub.47N.sub.6O.sub.4: 699.37 m/z Found 699.33
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.18 (s,
1H), 8.11 (s, 1H), 7.96-7.93 (m, 2H), 7.85-7.80 (m, 6H), 7.45 (br.
s, 1H), 7.37 (br. s, 1H), 5.36 (br. s, 2H), 3.72 (br. s, 1H), 3.64
(br. s, 1H), 2.82 (br. s, 1H), 2.72 (br s, 1H), 2.43 (br. s, 1H),
2.15 (br. s, 1H), 1.70 (br. s, 2H), 1.55/1.33 (s, 18H), 1.10 (br.
s, 1H), 0.94 (br. s, 1H), 0.88 (br. s, 1H), 0.81 (br. s, 1H).
##STR00573##
Intermediate 203
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-Azabicyclo[3.1.0]hex-3-yl)-1H-imida-
zol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
[0872] A solution of 4N HCl in dioxane (10 mL) was added to
tert-butyl
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-(tert-butoxycarbonyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2--
azabicyclo[3.1.0]hexane-2-carboxylate (375 mg. 0.537 mmol) in MeOH
(3 mL) and stirred at ambient conditions for 2 hours, concentrated,
and dried under vacuum. Tetra HCl salt (assume theoretical: 346
mg). LC-MS retention time 2.75 min; calcd. for
C.sub.32H.sub.31N.sub.6: 499.26 m/z Found 499.22 [M+H].sup.+. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% methanol/95% water/0.1% TFA and
Solvent B was 95% methanol/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00574##
[0873] Intermediate 209 was prepared as shown in Scheme 55
utilizing as starting material
(1S,3S,5S)-2-(tert-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-c-
arboxylic acid (which could be prepared according to WO
2004/052850). Analytical data for the intermediates shown
below.
##STR00575##
Intermediate 204
[0874] To a solution of the diastereomeric mixture (at the carbon
carrying the methyl group) of Intermediate 197 (4.75 g, 18.46 mmol)
was added superhydride (19.20 mL, 19.20 mmol) dropwise at
-50.degree. C. in a dry ice/acetone bath over 10 min. Hunig's base
(13.58 mL, 78 mmol) was added, stirred for 10 min, DMAP (0.122 g,
0.997 mmol) was added as a solid, stirred for 15 min, and
trifluoroacetic anhydride (2.98 mL, 21.08 mmol) was added dropwise
over 15 mins. The dry ice/acetone bath was removed and the reaction
mixture was stirred for 4 h while being allowed to warm to room
temperature. The reaction mixture was washed with water (50 mL),
sat. NaCl (30 mL), and concentrated in vacuo. The resulting crude
material was purified by flash chromatography (8-60% EtOAc/Hexane)
to afford ester Intermediate 204 as a yellow oil (2.85 g). .sup.1H
NMR (CDCl.sub.3, 400 MHz): 6.36 (s, 0.5H), 6.25 (s, 0.5H),
4.70-4.57 (m, 1H), 3.78 (s, 3H), 2.96 (m, 1H), 2.54 (m, 1H), 1.70
(s, 3H), 1.50 (s, 4.5H), 1.44 (s, 4.5H).
##STR00576##
Intermediate 205 and Intermediate 206
[0875] Diethylzinc (1.1 M in toluene, 59.1 mL, 65.0 mmol) was added
dropwise over 20 min to a cooled (-23.degree. C.) toluene (60 mL)
solution of Intermediate 203 (5.23 g, 21.68 mmol), and stirred for
10 min. Chloroiodomethane (9.44 mL, 130 mmol) was added dropwise
over 10 min, and the reaction mixture was stirred at -21.degree. C.
for 16 hr. Sat. NaHCO.sub.3 (60 mL) was added to the reaction
mixture, the cooling bath was removed, and the mixture was stirred
for 10 min. It was then filtered, and the filter cake was washed
with toluene (50 mL). The filterate was partitioned, and the
organic layer was dried with Na.sub.2SO.sub.4, and concentrated in
vacuo. The resulting crude material was purified with flash
chromatography (2-10% EtOAc/Hexane) to afford Intermediate 205
(first elute; colorless oil; 2.88 g) and Intermediate 206 (second
elute; colorless oil; 1.01 g). Relative stereochemical assignment
was made based on NOE studies. Intermediate 205: .sup.1H NMR
(CDCl.sub.3, 400 MHz): 4.65-4.52 (m, 1H), 3.72 (s, 3H), 3.28-3.17
(m, 1H), 2.44-2.32 (m, 1H), 2.16-2.10 (m, 1H), 1.51-1.42 (two s,
9H), 1.24 (s, 3H), 1.07 (m, 1H), 0.69-0.60 (m, 1H).
##STR00577##
Intermediate 207
(1S,3S,5S)-2-(tert-Butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-ca-
rb oxylic acid
[0876] To a solution of Intermediate 205 (2.88 g, 11.28 mmol) in
ethanol (20 mL) was added a solution of LiOH (0.324 g, 13.54 mmol)
in water (10.00 mL), and the mixture was stirred at room
temperature for 6 hr. Most of the volatile component was removed in
vacuo, and the residue was partitioned between water (20 mL) and
ether (20 mL). The aqueous layer was chilled in an ice-water bath,
acidified with a 1N HCl to a pH region of 2, and extracted with
EtOAc (30 mL, 4.times.). The combined organic phase was dried with
Na.sub.2SO.sub.4, evaporated in vacuo to give Intermediate 207 as a
sticky solid (2.55 g). .sup.1H NMR (CDCl.sub.3, 400 MHz): 4.64 (m,
1H), 3.25 (appt s, 1H), 2.70-2.40 (m, 1H), 2.14 (m, 1H), 1.54-1.44
(m, 9H), 1.27 (s, 3H), 1.10-0.80 (m, 1H), 0.67 (m, 1H).
##STR00578##
Intermediate 208
tert-Butyl
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-(tert-butoxycarbonyl)-5--
methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1-
H-imidazol-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0877] Hunig's base (0.475 mL, 2.72 mmol) was added to a stirred
solution of
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone
(0.607 g, 1.36 mmol) and
(1S,3S,5S)-2-(tert-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-c-
arboxylic acid (729.0 mg, 2.72 mmol) in dry acetonitrile (12 mL).
The heterogeneous mixture was stirred for 20 h at RT the solvent
was removed in vacuo. The residue was taken up in ethyl acetate and
washed with saturated sodium bicarbonate solution, brine, and dried
over sodium sulfate. Wash was repeated 2.times.. The crude product
was taken up in xylene (12 mL) and ammonia acetate (1.57 g, 20.39
mmol) was added. The pressure vessel was sealed and placed into a
preheated oil bath (140.degree. C.) and stirred for 2 h. The
solvent was removed in vacuo and the residue was taken up in ethyl
acetate and washed with saturated sodium bicarbonate solution and
brine. After being concentrated, the crude product was charged
(methylene chloride) to a 80 g Thompson silica gel cartridge
(eluted with 25% B to 100% B over 1.5 L and hold 0.5 L B. Solvent
B=ethyl acetate and Solvent A=hexanes) to yield tert-butyl
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-(tert-butoxycarbonyl)-5-methyl-2-a-
zabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (279 mg,
27%) as a light yellow solid. LC-MS retention time 3.56 min; calcd.
for C.sub.44H.sub.51N.sub.6O.sub.4: 727.40 m/z Found 727.41
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.18 (s,
1H), 8.11 (s, 1H), 7.96-7.93 (m, 2H), 7.83-7.80 (m, 6H), 7.43 (br.
s, 1H), 7.35 (br. s, 1H), 5.35 (br. s, 2H), 3.42 (br. s, 1H), 3.36
(br. s, 1H), 2.61 (br. s, 1H), 2.53 (br.s, 2H), 2.72 (br s, 1H),
1.55/1.33 (s, 25H), 1.06 (br. s, 1H), 0.85 (br. s, 1H), 0.72 (br.
s, 1H).
##STR00579##
Intermediate 209
(1S,3S,5S)-5-Methyl-3-(4-(4-(6-(2-((1S,3S,5S)-5-methyl-2-azabicyclo[3.1.0]-
hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexane
[0878] A solution of 4N HCl in dioxane (15 mL) was added to
tert-butyl
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-(tert-butoxycarbonyl)-5-methyl-2-a-
zabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (250 mg.
0.344 mmol) in MeOH (5 mL) and stirred at ambient conditions for 2
hours, concentrated, and dried under vacuum. Tetra HCl salt (assume
theoretical: 231 mg). LC-MS retention time 3.09 min; calcd. for
C.sub.34H.sub.35N.sub.6: 527.29 m/z Found 527.35 [M+H].sup.+. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% methanol/95% water/0.1% TFA and
Solvent B was 95% methanol/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00580##
[0879] Intermediate 212 was prepared as shown in Scheme 56
utilizing
(1R,3S,5R)-2-(tert-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-c-
arboxylic acid (which could be prepared according to WO 2004/052850
as starting material. Analytical data for the intermediates shown
below.
##STR00581##
Intermediate 210
(1R,3S,5R)-2-(tert-Butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-ca-
rboxylic acid
[0880] Intermediate 210 was prepared from Intermediate 206 as
described above for Intermediate 207. .sup.1H NMR (CDCl.sub.3, 400
MHz): 4.13 (app br s, 1H), 3.06 (app br s, 1H), 2.55/2.41
(overlapping app br s, 2H), 1.51 (s, 9H), 1.27 (s, 3H), 0.76 (app
t, J=5.6, 1H), 0.60 (app br s, 1H).
##STR00582##
Intermediate 211
tert-Butyl
(1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-5--
methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1-
H-imidazol-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate
[0881] Hunig's base (0.29 mL, 1.66 mmol) was added to a stirred
solution of
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone
(0.37 g, 0.83 mmol) and
(1R,3R,5S)-2-(tert-butoxycarbonyl)-5-methyl-2-azabicyclo[3.1.0]hexane-3-c-
arboxylic acid (0.40 g, 1.66 mmol) in dry acetonitrile (9 mL). The
heterogeneous mixture was stirred for 20 h at RT the solvent was
removed in vacuo. The residue was taken up in ethyl acetate and
washed with saturated sodium bicarbonate solution, brine, and dried
over sodium sulfate. Wash was repeated 2.times.. The crude product
was taken up in xylene (9 mL) and ammonia acetate (1.3 g, 16.6
mmol) was added. The pressure vessel was sealed and placed into a
preheated oil bath (140.degree. C.) and stirred for 2 h. The
solvent was removed in vacuo and the residue was taken up in ethyl
acetate and washed with saturated sodium bicarbonate solution and
brine. After being concentrated, the crude product was charged
(methylene chloride) to a 90 g Thompson silica gel cartridge
(eluted with 25% B to 100% B over 1.5 L and hold 0.5 L B. Solvent
B=ethyl acetate and Solvent A=hexanes) to yield tert-butyl
(1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-5-methyl-2-a-
zabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (80 mg,
10%). LC-MS retention time 3.37 min; calcd. for
C.sub.44H.sub.51N.sub.6O.sub.4: 727.40 m/z Found 727.38
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.19 (s,
1H), 8.08 (s, 1H), 7.92-7.91 (m, 2H), 7.85-7.77 (m, 6H), 7.47 (s,
1H), 7.38 (s, 1H), 4.67 (br. s, 2H), 3.93-3.90 (m, 2H), 2.63-2.57
(m, 2H), 2.20-2.14 (m, 2H), 1.31-1.23 (m, 24H), 0.92 (app. d, J=6.7
Hz, 1H), 0.82 (br. s, 1H), 0.77-0.75 (m, 2H).
##STR00583##
Intermediate 212
(1R,3S,5R)-5-Methyl-3-(4-(4-(6-(2-((1R,3S,5R)-5-methyl-2-azabicyclo[3.1.0]-
hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicy-
clo[3.1.0]hexane
[0882] A solution of 4N HCl in dioxane (5 mL) was added to
tert-butyl
(1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-5-methyl-2-a-
zabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate (48 mg.
0.066 mmol) in MeOH (1 mL) and stirred at ambient conditions for 2
hours, concentrated, and dried under vacuum. Tetra HCl salt (assume
theoretical: 44 mg). LC-MS retention time 2.82 min; calcd. for
C.sub.34H.sub.35N.sub.6: 527.29 m/z Found 527.26 [M+H].sup.+. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% methanol/95% water/0.1% TFA and
Solvent B was 95% methanol/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00584##
[0883] Intermediate 218 was prepared as shown in Scheme 57
utilizing
(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid (Intermediate 194) as starting material. Analytical data for
the intermediates shown below:
##STR00585##
Intermediate 213
(2S,4S)-tert-Butyl
2-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-4-methylpyrrolidine-1-car-
boxylate
[0884] Hunig's base (1.6 mL, 9.0 mmol) was added to a stirred
mixture of 2-bromo-1-(6-bromonaphthalen-2-yl)ethanone (3.0 g, 9.0
mmol) and
(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid (2.062 g, 8.99 mmol) in dry acetonitrile (100 mL) and stirred
at rt for 16 h. Additional
(S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic acid
(500 mg) and Hunig's base (0.4 mL) were added and stirring was
continued for 15 h. The solvent was removed by rotary evaporation
and the residue was taken up in EtOAc and washed with sat'd
NaHCO.sub.3 soln, brine, and dried (Na.sub.2SO.sub.4). The ester
was taken up in xylene (100 mL) and treated with NH.sub.4OAc (10.40
g, 135 mmol) in a sealed pressure vessel which was immersed in an
oil bath at 140.degree. C. and stirred for 2 h. The solvent was
removed by rotary evaporation under high vacuum and the residue was
taken up in EtOAc and washed with sat'd NaHCO.sub.3 soln, brine,
and dried (Na.sub.2SO.sub.4).
[0885] This residue was purified with a Thompson 300 g silica gel
cartridge (gradient elution 3% B to 100% B over 4.5 L and hold 0.5
L B; Solvent B=ethyl acetate and Solvent A=hexanes) to yield
(2S,4S)-tert-butyl
2-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-4-methylpyrrolidine-1-car-
boxylate (2 g) as a foam. For the purpose of characterization,
approximately 25 mg was dissolved in methanol (2 mL) and subjected
to prep HPLC (gradient 0-100% B over a 25 min at 40 mL/min) on a
PHENOMENEX.RTM.Luna column (30.times.100 mm, 10u); Solvent B=95%
CH.sub.3CN-5% H.sub.2O-10 mM NH.sub.4OAc and A=5% CH.sub.3CN-95%
H.sub.2O-10 mM NH.sub.4OAc. LC-MS retention time 2.61 min; calcd.
for C.sub.23H.sub.27BrN.sub.3O.sub.2: 456.13 m/z; found 456.09
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.18 (s,
1H), 8.03 (s, 1H), 7.89 (d, J=8.6 Hz, 1H), 7.82 (d, J=8.6 Hz, 1H),
7.79 (d, J=8.6 Hz, 1H), 7.57 (d, J=10.4 Hz, 1H), 7.51 (s, 1H), 4.91
(br. s, 1H), 3.81 (t, J=8.3 Hz, 1H), 3.21 (t, J=8.3 Hz, 1H),
2.53-2.48 (m, 1H), 2.37-2.34 (m, 1H), 1.81-1.73 (m, 1H), 1.46/1.21
(s, 9H), 1.16 (d, J=6.1 Hz, 3H).
##STR00586##
Intermediate 214
(2S,4S)-tert-Butyl
2-(4-(6-bromonaphthalen-2-yl)-5-chloro-1H-imidazol-2-yl)-4-methylpyrrolid-
ine-1-carboxylate
[0886] NCS (203 mg, 1.52 mmol) was added to a solution of
(2S,4S)-tert-butyl
2-(5-(6-bromonaphthalen-2-yl)-1H-imidazol-2-yl)-4-methylpyrrolidine-1-car-
boxylate (560 mg, 1.39 mmol) in dry DMF (15 mL) and the mixture was
heated at 50.degree. C. for 16 h. The solvent was removed by
nitrogen purge and the residue was purified with a Thompson 90 g
silica gel cartridge (gradient elution 5%-100% B over 2 L; Solvent
B=ethyl acetate and Solvent A=hexanes) to yield (2S,4S)-tert-butyl
2-(4-(6-bromonaphthalen-2-yl)-5-chloro-1H-imidazol-2-yl)-4-methylpyrrolid-
ine-1-carboxylate (255 mg) as a tan foam. For purpose of
characterization, approximately 20 mg was dissolved in methanol (2
mL) and subject to prep HPLC (gradient 0-100% B over a 25 min at 40
ml/min) on a PHENOMENEX.RTM. Luna column (30.times.100 mm, 10u);
Solvent B=95% CH.sub.3CN-5% H.sub.2O-10 mM NH.sub.4OAc and A=5%
CH.sub.3CN-95% H.sub.2O-10 mM NH.sub.4OAc. LC-MS retention time
2.61 min; calcd. for C.sub.23H.sub.26BrClN.sub.3O.sub.2: 492.09
m/z; found 492.03 [M+H]. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
methanol/95% water/0.1% TFA and Solvent B was 95% methanol/5%
water/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.18 (s, 1H), 8.09 (s, 1H), 7.91 (br. s, 2H), 7.83 (d,
J=8.6 Hz, 1H), 7.63 (d, J=8.6 Hz, 1H), 4.79-4.75 (m, 1H), 3.78 (t,
J=8.9 Hz, 1H), 3.19 (t, J=10.4 Hz, 1H), 2.49 (quin, J=6.1 Hz, 1H),
2.36-2.32 (m, 1H), 1.78 (q, J=11.9 Hz, 1H), 1.45/1.24 (s, 9H), 1.15
(d, J=6.1 Hz, 3H).
##STR00587##
Intermediate 215
(2S,4S)-tert-Butyl
2-(5-(4-bromophenyl)-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate
[0887] Hunig's base (2.3 mL, 13.1 mmol) was added to a stirred
mixture of 2-bromo-1-(4-bromophenyl)ethanone (3.64 g, 13.1 mmol)
and
(2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylic
acid (3.0 g, 13 mmol) in dry acetonitrile (150 mL) and stirred at
rt for 16 h. The solvent was removed by rotary evaporation and the
residue was taken up in EtOAc and washed with water, sat'd
NaHCO.sub.3 soln, brine, and dried (Na.sub.2SO.sub.4). The
resulting ester was taken up in xylene (150 mL) and treated with
NH.sub.4OAc (15.1 g, 196 mmol) in a sealed pressure vessel which
was immersed in an oil bath at 140.degree. C. and stirred for 2 h.
The solvent was removed by rotary evaporation under high vacuum and
the residue was taken up in EtOAc and washed with sat'd NaHCO.sub.3
soln, brine, and dried (Na.sub.2SO.sub.4), filtered and
concentrated. The crude product was purified with a Thompson 300 g
silica gel cartridge (gradient elution 3% B to 100% B over 4.5 L
and hold 0.5 L B; Solvent B=ethyl acetate and Solvent A=hexanes) to
yield (2S,4S)-tert-butyl
2-(5-(4-bromophenyl)-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate
(3.5 g, 59%) as a foam. For purpose of characterization,
approximately 25 mg was dissolved in methanol (2 mL) and subject to
prep HPLC (gradient 0-100% B over a 25 min at 40 ml/min) on a
PHENOMENEX.RTM. Luna column (30.times.100 mm, 10u); Solvent B=95%
CH.sub.3CN-5% H.sub.2O-10 mM NH.sub.4OAc and A=5% CH.sub.3CN-95%
H.sub.2O-10 mM NH.sub.4OAc. LC-MS retention time 3.06 min; calcd.
for C.sub.19H.sub.25BrN.sub.3O.sub.2: 408.11 m/z; found 408.16
[M+H]. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% methanol/95%
water/0.1% TFA and Solvent B was 95% methanol/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.62 (d,
J=8.2 Hz, 2H), 7.51 (d, J=8.6 Hz, 2H), 7.38 (s, 1H), 4.84-4.81 (m,
1H), 3.79 (t, J=8.6 Hz, 1H), 3.17 (t, J=10.4 Hz, 1H), 2.50-2.45 (m,
1H), 2.36-2.31 (m, 1H), 1.76-1.69 (m, 1H), 1.45/1.20 (s, 9H), 1.14
(d, J=6.1 Hz, 3H).
##STR00588##
Intermediate 216
(2S,4S)-tert-Butyl
4-methyl-2-(5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H--
imidazol-2-yl)pyrrolidine-1-carboxylate
[0888] Pd(Ph.sub.3P).sub.4 (213 mg, 0.185 mmol) was added to a
nitrogen purged suspension of (2S,4S)-tert-butyl
2-(5-(4-bromophenyl)-1H-imidazol-2-yl)-4-methylpyrrolidine-1-carboxylate,
(1.5 g, 3.7 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.88
g, 7.38 mmol) and potassium acetate (906 mg, 9.23 mmol) in dioxane
(35 mL). The pressure reaction vessel was degassed, sealed, and
immersed in an oil bath at 85.degree. C. for 16 hours. The mixture
was diluted with EtOAc and washed with sat'd NaHCO.sub.3 soln,
brine, and dried (Na.sub.2SO.sub.4), filtered and concentrated. The
crude product was purified with a Thompson 160 g silica gel
cartridge (gradient elution 10% to 100% B over 1 L. Solvent B=ethyl
acetate and Solvent A=hexanes) to yield (2S,4S)-tert-butyl
4-methyl-2-(5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H--
imidazol-2-yl)pyrrolidine-1-carboxylate (1.8 g, 97%) as a foam.
LC-MS retention time 1.78 min; calcd. for
C.sub.19H.sub.27BN.sub.3O.sub.4: 372.21 m/z; found 372.18 [M+H]. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% methanol/95% water/0.1% TFA and
Solvent B was 95% methanol/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00589##
Intermediate 217
(2S,4S)-tert-Butyl
2-(5-(6-(4-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1-
H-imidazol-5-yl)phenyl)naphthalen-2-yl)-4-chloro-1H-imidazol-2-yl)-4-methy-
lpyrrolidine-1-carboxylate
[0889] Pd(Ph.sub.3P).sub.4 (27.1 mg, 0.023 mmol) was added to a
nitrogen purged suspension of (2S,4S)-tert-butyl
2-(5-(6-bromonaphthalen-2-yl)-4-chloro-1H-imidazol-2-yl)-4-methylpyrrolid-
ine-1-carboxylate, (230 mg, 0.469 mmol), (2S,4S)-tert-butyl
4-methyl-2-(5-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H--
imidazol-2-yl)pyrrolidine-1-carboxylate (234 mg, 0.515 mmol) and
NaHCO.sub.3 (197 mg, 2.34 mmol) in an argon-degassed solution of
DME (4 mL) and water (1 mL). The pressure vessel was charged with
argon, sealed, and immersed into a pre-heated oil bath (85.degree.
C.) and stirred for 14 h. The mixture was diluted with EtOAc (Note:
THF and MeOH were added to aid dissolution) and washed with sat'd
NaHCO.sub.3 soln, brine, dried (Na.sub.2SO.sub.4), filtered and
concentrated. The residue was purified utilizing a Thompson 90 g
SiO.sub.2 column (Gradient elution 5% to 100% B for 2 L and hold
for 0.5 L Solvent B=ethyl acetate and Solvent A=hexanes) to yield
(2S,4S)-tert-butyl
2-(5-(6-(4-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1-
H-imidazol-5-yl)phenyl)naphthalen-2-yl)-4-chloro-1H-imidazol-2-yl)-4-methy-
lpyrrolidine-1-carboxylate (270 mg, 70%) as a foam. For the purpose
of characterization approx. 20 mg was dissolved in MeOH (up to 2
mL) and subject to prep HPLC vial (0 to 100% B over a 30 min
gradient at 40 ml/min) using a PHENOMENEX.RTM. Luna column
(30.times.100 mm, S10) where B=95% CH.sub.3CN-5% H.sub.2O-10 mM
NH.sub.4OAc and A=5% CH.sub.3CN-95% H.sub.2O-10 mM NH.sub.4OAc.
LC-MS retention time 3.90 min; calcd. for
C.sub.42H.sub.50ClN.sub.6O.sub.4: 737.36 m/z; found 737.43 [M+H].
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% methanol/95% water/0.1% TFA and
Solvent B was 95% methanol/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.21 (s, 1H), 8.16
(s, 1H), 8.03 (d, J=8.6 Hz, 1H), 7.99 (d, J=8.6 Hz, 2H), 7.91-7.81
(m, 6H), 7.42 (s, 1H), 4.86 (under MeOH, 1H), 4.80-4.877 (m, 1H),
3.82-3.77 (m, 2H), 3.20 (t, J=10.4 Hz, 2H), 2.52-2.47 (m, 2H),
2.37-2.34 (m, 2H), 1.83-1.73 (m, 2H), 1.46/1.26/1.23 (s, 18H), 1.16
(m, 6H).
##STR00590##
Intermediate 218
4-Chloro-2-((2S,4S)-4-methylpyrrolidin-2-yl)-5-(6-(4-(2-((2S,4S)-4-methylp-
yrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-1H-imidazole
[0890] 4N HCl in dioxane (5 mL) was added to a stirred solution of
(2S,4S)-tert-butyl
2-(5-(6-(4-(2-((2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidin-2-yl)-1-
H-imidazol-5-yl)phenyl)naphthalen-2-yl)-4-chloro-1H-imidazol-2-yl)-4-methy-
lpyrrolidine-1-carboxylate (30 mg, 0.041 mmol) in MeOH (1 mL) and
the mixture was stirred at RT for 2 h, concentrated, and dried
under high vacuum for 2 h. LC-MS retention time 3.31 min; calcd.
for C.sub.32H.sub.34ClN.sub.6: 537.26 m/z Found 537.37 [M+H]. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% methanol/95% water/0.1% TFA and
Solvent B was 95% methanol/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00591##
##STR00592##
Intermediate 219
(S)-Methyl 4,5-dihydro-1H-pyrazole-5-carboxylate
[0891] DMAP (2.83 g, 23.2 mmol) was added to a solution of
(S)-methyl 4,5-dihydro-1H-pyrazole-5-carboxylate (prepared
according to J. Am. Chem. Soc., 119:8379-8380 (1997); 2.95 g, 23.02
mmol) and Boc.sub.2O (11.9 mL, 51.2 mmol) in CH.sub.2Cl.sub.2 (40
mL) and stirred at ambient condition for 22.5 hr. Additional
Boc.sub.2O (1.83 g) was added and stirring was continued for 15 hr.
Silica gel was added to the reaction mixture and the solvent was
removed in vacuo, and the resultant mesh was submitted to a
BIOTAGE.RTM. purification (300 g silica gel; column was eluted with
30-50% EtOAc/hexanes) to afford (S)-methyl
4,5-dihydro-1H-pyrazole-5-carboxylate as a yellow oil (3.956 g). A
sample of the starting pyrazoline, contaminated with the product,
was also retrieved (695 mg). .sup.1H NMR (CDCl.sub.3, .delta.=7.24
ppm, 400 MHz): 6.80 (s, 1H), 4.67 (dd, J=12.6, 6.1 Hz, 1H), 3.75
(s, 3H), 3.22 (ddd, J=18.5, 12.6, 1.4 Hz, 1H), 2.94 (ddd, J=18.5,
6.1, 1.7 Hz, 1H), 1.5 (s, 9H). LC-MS: Anal. Calcd. for [M+Na].sup.+
C.sub.10H.sub.16N.sub.2NaO.sub.4: 251.10; found 251.26.
##STR00593##
Intermediate 220
(S)-1-tert-Butyl 5-methyl 1H-pyrazole-1,5(4H,5H)-dicarboxylate
[0892] Sodium cyanoborohydride (0.769 g, 12.2 mmol) was added in
batches over 1 min to an acetic acid (7.0 mL) solution of
(S)-methyl 4,5-dihydro-1H-pyrazole-5-carboxylate (1.05 g, 4.61
mmol), and stirred at ambient condition for 20 h. Formaldehyde (1
mL of 37% in water) was added dropwise over 4 min, and stirring was
continued for 4.5 h. The volatile component was removed in vacuo
and the residue was treated with saturated NaHCO.sub.3 (10 mL) and
CH.sub.2Cl.sub.2 (30 mL), the mixture was shaken and the phases
were separated. The organic layer was washed with an additional
saturated NaHCO.sub.3 solution (10 mL), dried (MgSO.sub.4),
filtered and concentrated in vacuo. The resultant crude material
was purified with a BIOTAGE.RTM. (240 g silica gel; sample was
loaded with CH.sub.2Cl.sub.2; eluted with 60-100% EtOAc/hexanes) to
afford (S)-1-tert-butyl 5-methyl
1H-pyrazole-1,5(4H,5H)-dicarboxylate as a colorless oil (861 mg).
.sup.1H NMR (CDCl.sub.3, .delta.=7.24 ppm, 400 MHz): 4.39 (app br
t, J=7.7, 1H), 3.74 (s, 3H), 3.15-3.02 (m, 2H), 2.64 (s, 3H),
2.48-2.40 (m, 1H), 2.30-2.21 (m, 1H), 1.45 (s, 9H). LC-MS: Anal.
Calcd. for [M+Na].sup.+ C.sub.11H.sub.20N.sub.2NaO.sub.4: 267.13;
found 267.28.
##STR00594##
Intermediate 221
Lithium (S)-2-(tert-butoxycarbonyl)pyrazolidine-3-carboxylate
[0893] A water (5 mL) solution of LiOH (0.146 g, 6.09 mmol) was
added to a methanol (5 mL) solution of (S)-1-tert-butyl 5-methyl
1H-pyrazole-1,5(4H,5H)-dicarboxylate (0.76 g, 3.11 mmol), and
stirred at ambient conditions for .about.7 h. The reaction mixture
was cooled with an ice-water bath, and HCl/H.sub.2O (3 mL of 1.00
N, 3.0 mmol) was added dropwise and stirred for a few minutes.
Then, the volatile component was removed in vacuo and the resultant
viscous oil was exposed to high vacuum to afford lithium
(S)-2-(tert-butoxycarbonyl)pyrazolidine-3-carboxylate as a white
foam, which was used without further purification. .sup.1H NMR
(DMSO-d.sub.6,.delta.=2.50 ppm, 400 MHz) for crude sample: 3.98
(dd, J=8.5, 6.3, 1H), 2.86-2.74 (m, 2H), 2.45 (s, 3H), 2.25-2.17
(m, 1H), 2.10-2.02 (m, 1H), 1.35 (s, 9H). LC-MS: Anal. Calcd. for
[M+H].sup.+ C.sub.10H.sub.19N.sub.2O.sub.4: 231.13; found
231.21.
##STR00595##
##STR00596##
Intermediate 222
2-Amino-1-(4-(6-(2-aminoacetyl)naphthalen-2-yl)phenyl)ethanone
[0894] A slurry of
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (1.0
g, 2.2 mmol) and hexamethylenetetramine (0.660 g, 4.71 mmol) were
slurried into chloroform (30 mL) and heated at 50.degree. C. for 2
h. The reaction was cooled to rt and the solids were collected by
filtration (rinsed with chloroform, 3.times.25 mL). The resulting
bright yellow solid was slurried into MeOH (15.0 mL) and then conc
HCl (3.0 mL, 99 mmol) was added dropwise and the reaction was
stirred in the dark for 2 d. The reaction was concentrated to a
solid which was slurried into water and stirred for 1 h. The solids
were collected by filtration and the cake was dried in a vacuum
oven to yield an HCl salt of
2-amino-1-(4-(6-(2-aminoacetyl)naphthalen-2-yl)phenyl)ethanone (787
mg). LC-MS retention time 2.342 min; m/z 319.09 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00597##
Intermediate 223
(S)-tert-Butyl
5-(2-(4-(6-(2-((S)-2-(tert-butoxycarbonyl)-1-methylpyrazolidine-3-carboxa-
mido)acetyl)naphthalen-2-yl)phenyl)-2-oxoethylcarbamoyl)-2-methylpyrazolid-
ine-1-carboxylate
[0895] HATU (1577 mg, 4.15 mmol) was added to a stirred solution of
a n HCl salt of
2-amino-1-(4-(6-(2-aminoacetyl)naphthalen-2-yl)phenyl)ethanone (773
mg, 1.98 mmol) and lithium
(S)-2-(tert-butoxycarbonyl)-1-methylpyrazolidine-3-carboxylate (980
mg, 4.15 mmol) in DMF (15 mL) and Hunig's Base (2.070 mL, 11.85
mmol) and the reaction mixture was stirred for 2 h. The reaction
was concentrated to .about.20% volume, diluted with MeOH, filtered
and purified by HPLC (MeOH/water with an ammonium acetate buffer)
to yield (S)-tert-butyl
5-(2-(4-(6-(2-((S)-2-(tert-butoxycarbonyl)-1-methylpyrazolidine-3-carboxa-
mido)acetyl)naphthalen-2-yl)phenyl)-2-oxoethylcarbamoyl)-2-methylpyrazolid-
ine-1-carboxylate (537 mg) as a bright yellow solid. LC-MS
retention time 3.758 min; m/z 741.55 (MH-). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00598##
Intermediate 224
(S)-tert-Butyl
5-(5-(4-(6-(2-((S)-2-(tert-butoxycarbonyl)-1-methylpyrazolidin-3-yl)-1H-i-
midazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-methylpyrazolidin-
e-1-carboxylate
[0896] In a 25 mL pressure tube, (S)-tert-butyl
5-(2-(4-(6-(2-((S)-2-(tert-butoxycarbonyl)-1-methylpyrazolidine-3-carboxa-
mido)acetyl)naphthalen-2-yl)phenyl)-2-oxoethylcarbamoyl)-2-methylpyrazolid-
ine-1-carboxylate (420 mg, 0.565 mmol) and ammonium acetate (872
mg, 11.3 mmol) were slurried into xylenes (7 mL). The reaction
vessel was sealed and heated at 135.degree. C. for 5 h then at rt
ON. The crude reaction was diluted with 1/2 sat aq. NaHCO.sub.3 (40
mL) and extracted with DCM (3.times.30 mL). The combine organics
were dried (MgSO.sub.4), filtered and concentrated to yield
(S)-tert-butyl
5-(5-(4-(6-(2-((S)-2-(tert-butoxycarbonyl)-1-methylpyrazolidin-3-yl)-1H-i-
midazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-methylpyrazolidin-
e-1-carboxylate (95 mg, .about.80% purity) as a dark yellow glass.
LC-MS retention time 4.026 min; m/z 703.63 (MH-). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.32 (s,
1H), 8.28 (s, 1H), 8.15 (d, J=8.8 Hz, 1H), 8.10 (d, J=8.8 Hz, 1H),
7.86-8.03 (m, 8H), 5.31-5.39 (m, 2H), 3.22-3.35 (m, 4H), 2.82-2.93
(m, 2H), 2.77 (s, 3H), 2.76 (s, 3H), 2.53-2.65 (m, 2H), 1.43 (s,
18H).
##STR00599##
Condition 1
Column=PHENOMENEX.RTM., 2.0.times.50 mm, 3 .mu.m
Start % B=0
Final % B=100
[0897] Gradient time=4 min Stop time=5 min Flow Rate=0.8 mL/min
Wavelength=220 nm
[0898] Solvent A=0.1% TFA in 10% methanol/90% water Solvent B=0.1%
TFA in 90% methanol/10% water Oven temp.=40.degree. C.
Condition 2
Column=Sunfire, C18, 3.0.times.150 mm, 3.5 .mu.m
Start % B=0
Final % B=100
[0899] Gradient time=15 min Stop time=18 min Flow Rate=1 mL/min
Wavelength 1=220 nm
Wavelength 2=254 nm
[0900] Solvent A=0.1% TFA in 5% MeCN/95% water Solvent B=0.1% TFA
in 95% MeCN/5% water
##STR00600##
Intermediate 225
(S)-2-(4-(6-(2-((S)-2-(tert-Butoxycarbonylamino)-3,3-dimethylbutanoyloxy)a-
cetyl)naphthalen-2-yl)phenyl)-2-oxoethyl
2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoate
[0901] To a solution of
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone (0.2
g, 0.448 mmol) and
(S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoic acid (0.207
g, 0.897 mmol) in DCM was added DIPEA (0.172 mL, 0.986 mmol). The
reaction mixture was stirred at rt for 16 h and then diluted with
EtOAc, washed with sat. NaHCO.sub.3, water, sat. NaCl and dried
over anhydrous Na.sub.2SO.sub.4. Then it was filtered and
concentrated to yield
(S)-2-(4-(6-(2-((S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutano-
yloxy)acetyl)naphthalen-2-yl)phenyl)-2-oxoethyl
2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoate as a pale yellow
solid. LC-MS (Cond. 1): [M+H].sup.+747.50, Rt=4.67 min.
##STR00601##
Intermediate 226
tert-Butyl(S)-1-(5-(4-(6-(2-((S)-2,2-dimethyl-1-(tert-butoxycarbonylamino)-
propyl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2,2-dim-
ethylpropylcarbamate
[0902] In a sealed tube, a reaction mixture of
(S)-2-(4-(6-(2-((S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoyloxy)-
acetyl)naphthalen-2-yl)phenyl)-2-oxoethyl
2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoate (0.35 g, 0.469
mmol) in xylenes (5 mL) and ammonium acetate (0.361 g, 4.69 mmol)
was heated at 130.degree. C. for 3 h. The reaction mixture was
diluted with EtOAc and water. The organic layer was washed with
sat. NaHCO.sub.3 and sat. NaCl, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
charged into a 40 g silica gel cartridge which was eluted with a 15
min gradient of 0-100% EtOAc/Hex to yield
tert-butyl(S)-1-(5-(4-(6-(2-((S)-2,2-dimethyl-1-(tert-butoxycarbonylamino-
)propyl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2,2-di-
methylpropylcarbamate (0.2 g) as a yellow solid. LC-MS (Cond. 1):
[M+H].sup.+707.6, Rt=3.44 min.
##STR00602##
Intermediate 226
(S)-1-(5-(4-(6-(2-((S)-1-Amino-2,2-dimethylpropyl)-1H-imidazol-5-yl)naphth-
alen-2-yl)phenyl)-1H-imidazol-2-yl)-2,2-dimethylpropan-1-amine
[0903] To a reaction mixture of
tert-butyl(S)-1-(5-(4-(6-(2-((S)-2,2-dimethyl-1-(tert-butoxycarbonylamino-
)propyl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2,2-di-
methylpropylcarbamate (0.2 g, 0.283 mmol) in DCM was added 4M
hydrogen chloride in dioxane (2 mL, 8 mmol). The reaction mixture
was stirred at rt for 4 h and then concentrated to dryness to yield
an HCl salt of
(S)-1-(5-(4-(6-(2-((S)-1-amino-2,2-dimethylpropyl)-1H-imidazol-5-yl)napht-
halen-2-yl)phenyl)-1H-imidazol-2-yl)-2,2-dimethylpropan-1-amine as
a yellow solid. LC-MS (Cond. 1): [M+H].sup.+507.43, Rt=3.241
min.
EXAMPLES
##STR00603##
[0904] Example 1
Methyl
((1S)-1-(((2S)-2-(5-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-ben-
zimidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0905] HATU (59 mg, 0.15 mmol) was added to a mixture of a TFA salt
of
2,7-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(37 mg), diisopropylethylamine (68 .mu.L, 0.40 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (27 mg, 0.15
mmol) in DMF (2 mL) and the resulting mixture was stirred at
ambient conditions for 2.5 hours. The reaction mixture was diluted
with methanol (2 mL) and water (2 mL) and stirred for 15 min. The
volatile component was removed in vacuo and the residue was
purified twice by a reverse phase HPLC (water/acetonitrile/TFA) and
reverse phase HPLC (water/acetonitrile/NH.sub.4OAc) to provide the
desired product which was suspended in methanol and TFA. The
volatiles were removed in vacuo to afford a TFA salt of methyl
((1S)-1-(((2S)-2-(5-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimida-
zol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate (11.3
mg) as tan solid. LC-MS retention time 1.34 min; calcd. for
C.sub.46H.sub.52N.sub.8O.sub.6 812.4; found m/z 813.42 [M+H].sup.+.
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a Waters Sunfire 5u C18 4.6.times.50 mm column using
a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 3 min, a hold time of 1 min and an analysis time
of 4 min where Solvent A was 10% acetonitrile/90% water/0.1% TFA
and Solvent B was 90% acetonitrile/10% water/0.1% TFA. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.31 (s, 2H), 8.03-8.10 (m,
4H), 7.96-8.02 (m, Hz, 2H), 7.89 (dd, J=8.6, 1.5 Hz, 2H), 7.85 (m,
2H), 5.36 (t, J=7.0 Hz, 2H), 4.22-4.32 (m, 2H), 4.09-4.19 (m, 2H),
3.89-4.00 (m, 2H), 3.63-3.74 (m, 6H), 2.56-2.70 (m, 2H), 2.31-2.40
(m, 2H), 2.19-2.31 (m, 4H), 2.08 (dd, J=13.6, 6.9 Hz, 2H), 0.95 (d,
J=7.0 Hz, 6H), 0.89 (d, J=6.7 Hz, 6H).
##STR00604##
Example 2
Methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-ben-
zimidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0906] HATU (82 mg, 0.217 mmol) was added to a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(69 mg), diisopropylethylamine (126 .mu.L, 0.723 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (51 mg, 0.29
mmol) in DMF (2 mL) and the resulting mixture was stirred at
ambient conditions for 2.5 hours. The reaction mixture was diluted
with methanol (2 mL) and water (2 mL). Then the volatile component
was removed in vacuo and the residue was purified by a reverse
phase HPLC (water/acetonitrile/TFA) to provide a TFA salt of methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimida-
zol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate (47 mg)
as tan solid. LC-MS retention time 1.30 min; calcd. for
C.sub.46H.sub.52N.sub.8O.sub.6: 812.40; found m/z 813.42
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.25 (s, 2H),
8.07-8.16 (m, 4H), 8.02 (m, J=8.6 Hz, 2H), 7.91 (m, J=8.6 Hz, 2H),
7.87 (d, J=8.6 Hz, 2H), 5.37 (t, J=7.2 Hz, 2H), 4.28 (d, J=7.0 Hz,
2H), 4.09-4.18 (m, 2H), 3.90-4.01 (m, 2H), 3.63-3.73 (m, 6H),
2.57-2.70 (m, 2H), 2.32-2.45 (m, 2H), 2.19-2.32 (m, 4H), 2.09 (dq,
J=13.6, 6.9 Hz, 2H), 0.95 (d, J=6.7 Hz, 6H), 0.89 (d, J=6.7 Hz,
6H).
##STR00605##
Example 3
Methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-(1-((2S)-2-((methoxycarbonyl)amino)-3--
methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)phenyl)-1-
H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0907] HATU (39 mg, 0.10 mmol) was added to a mixture of a TFA salt
of
2-((S)-pyrrolidin-2-yl)-6-(6-(4-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl-
)phenyl)naphthalen-2-yl)-1H-benzo[d]imidazole (50 mg),
diisopropylethylamine (89 .mu.L, 0.51 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (27 mg, 0.15
mmol) in DMF (2 mL) and the resulting mixture was stirred at
ambient conditions for 2.5 hours. The reaction mixture was diluted
with methanol (2 mL) and water (2 mL). The volatile component was
removed in vacuo and the residue was purified by a reverse phase
HPLC (water/acetonitrile/TFA) to provide a TFA salt of methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-(1-((2S)-2-((methoxycarbonyl)amino)-3-methyl-
butanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)phenyl)-1H-imid-
azol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate (31.4
mg) as tan solid. LC-MS retention time 1.37 min; calcd. for
C.sub.48H.sub.54N.sub.8O.sub.6 838.42; found m/z 839.36
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.23-8.28 (m, 2H),
8.08-8.15 (m, 3H), 8.02 (d, J=8.6 Hz, 1H), 7.95-8.00 (m, 2H),
7.85-7.95 (m, 6H), 5.40 (t, J=7.2 Hz, 1H), 5.29 (t, J=7.5 Hz, 1H),
4.29 (dd, J=19.4, 7.2 Hz, 2H), 4.10-4.19 (m, 2H), 3.96-4.03 (m,
1H), 3.87-3.96 (m, 1H), 3.69 (m, 6H), 2.55-2.73 (m, 2H), 2.04-2.42
(m, 8H), 0.89-1.05 (m, 12H).
##STR00606##
Example 4
Methyl
((1S)-1-(((2S)-2-(4-(4-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)phen-
yl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0908] HATU (29 mg, 0.076 mmol) was added to a mixture of a TFA
salt of
2-((S)-pyrrolidin-2-yl)-6-(7-(4-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl-
)phenyl)naphthalen-2-yl)-1H-benzo[d]imidazole (25 mg),
diisopropylethylamine (45 .mu.L, 0.26 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (13 mg, 0.76
mmol) in DMF (2 mL) and the resulting mixture was stirred at
ambient conditions for 2.5 hours. The reaction mixture was diluted
with methanol (2 mL) and water (2 mL). The volatile component was
removed in vacuo and the residue was purified by a reverse phase
HPLC (water/acetonitrile/TFA) to provide a TFA salt of methyl
((1S)-1-(((2S)-2-(4-(4-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)phenyl)-1H-
-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
(14.2 mg) as yellow solid. LC-MS retention time 1.40 min; calcd.
for C.sub.48H.sub.54N.sub.8O.sub.6 838.42; found m/z 839.36
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.33 (br s,
2H), 8.13 (s, 1H), 8.03-8.10 (m, 3H), 8.00 (d, J=8.2 Hz, 2H),
7.86-7.94 (m, 6H), 5.40 (t, J=7.2 Hz, 1H), 5.29 (t, J=7.6 Hz, 1H),
4.31 (d, J=7.3 Hz, 1H), 4.27 (d, J=7.3 Hz, 1H), 4.10-4.20 (m, 2H),
3.85-4.10 (m, 2H), 3.69 (s, 6H), 2.55-2.74 (m, 2H), 2.03-2.37 (m,
8H), 0.85-1.05 (m, 12H).
##STR00607##
Example 5
Methyl
((1S)-2-((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-p-
yrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyr-
rolidinyl)-1-methyl-2-oxoethyl)carbamate
[0909] HATU (74.5 mg, 0.196 mmol) was added to a stirring solution
of a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(40 mg), (S)-2-(methoxycarbonylamino)propanoic acid (28.8 mg, 0.196
mmol) and DIEA (0.114 mL, 0.653 mmol) in DMF (2 mL). The reaction
was stirred 9 h at room temperature, diluted with MeOH (2 mL) and
water (2 mL), concentrated to remove the volatiles and purified by
preparative HPLC (acetonitrile/water with 0.1% TFA) to afford a TFA
salt of methyl
((1S)-2-((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrroli-
dinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidi-
nyl)-1-methyl-2-oxoethyl)carbamate (35.9 mg) as white solid. LC-MS
retention time 1.12 min; calcd. for C.sub.42H.sub.44N.sub.8O.sub.6
756.34; found m/z 757.20 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA. .sup.1H NMR (TFA salt,
500 MHz, MeOD) .delta. ppm 8.22-8.30 (m, 2H), 8.08-8.18 (m, 4H),
7.99-8.07 (m, 2H), 7.83-7.96 (m, 4H), 5.44 (dd, J=8.6, 5.2 Hz, 2H),
4.56 (q, J=6.9 Hz, 2H), 4.0-4.09 (m, 2H), 3.94-4.02 (m, 2H),
3.61-3.75 (m, 6H), 2.58-2.72 (m, 2H), 2.21-2.38 (m, 6H), 1.32-1.45
(m, 6H).
##STR00608##
Example 6
Dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrroli-
dinediyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate
[0910] HATU (74.5 mg, 0.196 mmol) was added to a stirring solution
of a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(40 mg), (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (41.0 mg,
0.196 mmol) and DIEA (0.114 mL, 0.653 mmol) in DMF (2 mL). The
reaction was stirred 9 h at room temperature, diluted with MeOH (2
mL) and water (2 mL), concentrated to remove the volatiles and
purified by preparative HPLC (acetonitrile/water with 0.1% TFA) and
then by preparative HPLC (acetonitrile/water with 10 mM ammonium
acetate) to afford dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrrolidinediyl-
((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate (32.3 mg) as
white solid. LC-MS retention time 1.43 min; calcd. for
C.sub.52H.sub.48N.sub.8O.sub.6 880.37; found m/z 881.23
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.25-8.35
(m, 2H), 8.12-8.21 (m, 4H), 8.05-8.12 (m, 2H), 7.90-8.00 (m, 4H),
7.38-7.54 (m, 10H), 5.62 (s, 2H), 5.38-5.53 (m, 2H), 4.04-4.18 (m,
2H), 3.67 (s, 6H), 2.34-2.41 (m, 2H), 2.46-2.58 (m, 2H), 2.15-2.35
(m, 4H), 2.02-2.13 (m, 2H).
##STR00609##
Example 7
5,5'-(2,6-Naphthalenediyl)bis(2-(2S)-1-((2R)-2-phenyl-2-(1-piperidinyl)ace-
tyl)-2-pyrrolidinyl)-1H-benzimidazole)
[0911] HATU (74.5 mg, 0.196 mmol) was added to a stirring solution
of a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(40 mg), (R)-2-phenyl-2-(piperidin-1-yl)acetic acid (43.0 mg, 0.196
mmol) and DIEA (0.114 mL, 0.653 mmol) in DMF (2 mL). The reaction
was stirred 9 h at room temperature, diluted with MeOH (2 mL) and
water (2 mL), concentrated to remove the volatiles and purified by
preparative HPLC (acetonitrile/water with 0.1% TFA) and then by
preparative HPLC (acetonitrile/water with 10 mM ammonium acetate)
to afford
5,5'-(2,6-naphthalenediyl)bis(2-(2S)-1-((2R)-2-phenyl-2-(1-piperidinyl)ac-
etyl)-2-pyrrolidinyl)-1H-benzimidazole) (44.3 mg) as white solid.
LC-MS retention time 1.18 min; calcd. for [M+H].sup.+
C.sub.58H.sub.50N.sub.8O.sub.2 900.48; found m/z 451.33 [1/2
M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA.
##STR00610##
Example 8
(1R)-2-((2S)-2-(5-(6-(2-((2S)-1-((2R)-2-(Dimethylamino)-2-phenylacetyl)-2--
pyrrolidinyl)-1H-benzimidazol-6-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-py-
rrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine
[0912] HATU (74.5 mg, 0.196 mmol) was added to a stirring solution
of a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(40 mg, 0.065 mmol), (R)-2-(dimethylamino)-2-phenylacetic acid
(35.1 mg) and DIEA (0.114 mL, 0.653 mmol) in DMF (2 mL). The
reaction was stirred 9 h at room temperature, diluted with MeOH (2
mL) and water (2 mL), concentrated to remove the volatiles and
purified by preparative HPLC (acetonitrile/water with 0.1% TFA) and
then by preparative HPLC (acetonitrile/water with 10 mM ammonium
acetate) to afford
(1R)-2-((2S)-2-(5-(6-(2-((2S)-1-((2R)-2-(dimethylamino)-2-phenylacetyl)-2-
-pyrrolidinyl)-1H-benzimidazol-6-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-p-
yrrolidinyl)-N,N-dimethyl-2-oxo-1-phenylethanamine (44.3 mg) as
off-white solid. LC-MS retention time 1.07 min; calcd. for
C.sub.52H.sub.52N.sub.8O.sub.2 820.42; found m/z 821.29
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.31 (s,
2H), 8.20-8.24 (m, 2H), 8.13-8.20 (m, 2H), 8.07-8.13 (m, 2H),
7.89-8.05 (m, 4H), 7.60-7.70 (m, 10H), 5.55-5.59 (m, 2H), 5.52 (dd,
J=8.7, 3.8 Hz, 2H), 4.11-4.19 (m, 2H), 2.42-3.18 (m, 13H),
2.17-2.36 (m, 5H), 1.98-2.08 (m, 4H).
##STR00611##
Example 9
Methyl
((1S,2R)-2-methoxy-1-(((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-
-O-methyl-L-threonyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-
-benzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate
[0913] HATU (74.5 mg, 0.196 mmol) was added to a stirring solution
of a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(40 mg), (2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid
(37.4 mg, 0.196 mmol) and DIEA (0.114 mL, 0.653 mmol) in DMF (2
mL). The reaction was stirred 9 h at room temperature, diluted with
MeOH (2 mL) and water (2 mL), concentrated to remove the volatiles
and purified by preparative HPLC (acetonitrile/water with 0.1% TFA)
and then by preparative HPLC (acetonitrile/water with 10 mM
ammonium acetate) to afford methyl
((1S,2R)-2-methoxy-1-(((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-O-met-
hyl-L-threonyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzi-
midazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate (9.3 mg) as
white solid. LC-MS retention time 1.22 min; calcd. for [M+H].sup.+
C.sub.46H.sub.52N.sub.8O.sub.8 844.39; found m/z 845.25
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.10-8.21 (m, 2H),
7.99-8.08 (m, 2H), 7.83-7.96 (m, 4H), 7.58-7.77 (m, 4H), 5.70-5.75
(m, 0.4H), 5.35 (dd, J=7.9, 4.9 Hz, 1.6H), 4.58-4.61 (m, 0.2H),
4.51 (d, J=4.9 Hz, 1.5H), 4.37-4.43 (m, 0.3H), 3.89-4.08 (m, 3H),
3.61-3.78 (m, 7H), 3.38-3.45 (m, 2H), 3.23-3.31 (m, 5H), 1.93-2.57
(m, 9H), 1.10-1.28 (m, 6H).
##STR00612##
Example 10
Methyl
((1S)-3-methoxy-1-(((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-O--
methyl-L-homoseryl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-b-
enzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate
[0914] HATU (74.5 mg, 0.196 mmol) was added to a stirring solution
of a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(40 mg), (S)-4-methoxy-2-(methoxycarbonylamino)butanoic acid (37.4
mg, 0.196 mmol) and DIEA (0.114 mL, 0.653 mmol) in DMF (2 mL). The
reaction was stirred 9 h at room temperature, diluted with MeOH (2
mL) and water (2 mL), concentrated to remove the volatiles and
purified by preparative HPLC (acetonitrile/water with 0.1% TFA) to
afford a TFA salt of methyl
((1S)-3-methoxy-1-(((2S)-2-(5-(6-(2-((2S)-1-(N-(methoxycarbonyl)-O-methyl-
-L-homoseryl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimi-
dazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)carbamate (28.6 mg) as
yellow solid. LC-MS retention time 1.18 min; calcd. for [M+H].sup.+
C.sub.46H.sub.52N.sub.8O.sub.8 844.39; found m/z 845.22
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.23-8.30
(m, 2H), 8.09-8.18 (m, 4H), 8.00-8.09 (m, 2H), 7.84-7.97 (m, 4H),
5.84-5.90 (m, 0.2H), 5.44 (dd, J=8.2, 4.9 Hz, 1.8H), 4.65 (dd,
J=9.2, 4.0 Hz, 1.8H), 4.38-4.45 (m, 0.2H), 3.91-4.11 (m, 4H),
3.66-3.78 (m, 6H), 3.39-3.55 (m, 4H), 3.27-3.38 (m, 7H), 2.59-2.75
(m, 2H), 2.25-2.40 (m, 5H), 2.16 (dddd, J=14.3, 7.0, 6.8, 4.3 Hz,
2H), 1.74-1.93 (m, 2H).
##STR00613##
Example 11
Dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrroli-
dinediyl((1S)-1-cyclopropyl-2-oxo-2,1-ethanediyl)))biscarbamate
[0915] HATU (74.5 mg, 0.196 mmol) was added to a stirring solution
of a TFA salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(40 mg, 0.065 mmol),
(S)-2-cyclopropyl-2-(methoxycarbonylamino)acetic acid (33.9 mg,
0.196 mmol) and DIEA (0.114 mL, 0.653 mmol) in DMF (2 mL). The
reaction was stirred 9 h at room temperature, diluted with MeOH (2
mL) and water (2 mL), concentrated to remove the volatiles and
purified by preparative HPLC (acetonitrile/water with 0.1% TFA) to
afford a TFA salt of dimethyl
(1S,1'S)-2,2'-((2S,2'S)-2,2'-(5,5'-(naphthalene-2,6-diyl)bis(1H-benzo[d]i-
midazole-5,2-diyl))bis(pyrrolidine-2,1-diyl))bis(1-cyclopropyl-2-oxoethane-
-2,1-diyl)dicarbamate (34.9 mg, 0.038 mmol, 58% yield) as white
solid. LC-MS retention time 1.23 min; calcd. for
C.sub.46H.sub.48N.sub.8O.sub.6 808.37; found m/z 809.22
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.20-8.29
(m, 2H), 8.07-8.16 (m, 4H), 7.98-8.07 (m, 2H), 7.84-7.95 (m, 4H),
5.37-5.48 (m, 2H), 4.03-4.13 (m, 2H), 3.87-4.03 (m, 4H), 3.64-3.74
(m, 6H), 2.65-2.74 (m, 2H), 2.20-2.40 (m, 6H), 1.09-1.22 (m, 2H),
0.47-0.67 (m, 6H), 0.30-0.46 (m, 2H).
##STR00614##
Example 12
Dimethyl
(2,7-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrroli-
dinediyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate
[0916] HATU (56.2 mg, 0.148 mmol) was added to a stirred solution
of a TFA salt of
2-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-5-yl)-7-(2-((S)-py-
rrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene (47 mg),
(R)-2-(methoxycarbonylamino)-2-phenylacetic acid (30.9 mg, 0.148
mmol) and DIEA (0.086 mL, 0.49 mmol) in DMF (2 mL). The reaction
was stirred 4 h at room temperature, diluted with MeOH (2 mL) and
water (2 mL) concentrated to remove the volatiles and purified by
preparative HPLC (acetonitrile/water with 0.1% TFA) to afford a TFA
salt of dimethyl
(1R,1'R)-2,2'-((2S,2'S)-2,2'-(5,5'-(naphthalene-2,7-diyl)bis(1H-benzo[d]i-
midazole-5,2-diyl))bis(pyrrolidine-2,1-diyl))bis(2-oxo-1-phenylethane-2,1--
diyl)dicarbamate (38 mg) as light yellow solid. LC-MS retention
time 1.45 min; calcd. for C.sub.52H.sub.48N.sub.8O.sub.6 880.37;
found m/z 881.26 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. .sup.1H NMR (500 MHz, MeOD)
.delta. ppm 8.31-8.39 (m, 2H), 8.03-8.17 (m, 6H), 7.84-8.00 (m,
4H), 7.33-7.53 (m, 10H), 5.55-5.63 (m, 2H), 5.40-5.54 (m, 2H),
4.01-4.17 (m, 2H), 3.67 (s, 6H), 3.27-4.41 (m, 2H), 2.44-2.75 (m,
2H), 2.01-2.33 (m, 6H).
##STR00615##
Example 13
Methyl
((1S)-2-((2S)-2-(5-(7-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-p-
yrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyr-
rolidinyl)-1-methyl-2-oxoethyl)carbamate
[0917] HATU (56.2 mg, 0.148 mmol) was added to a stirred solution
of a TFA salt of
2-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-5-yl)-7-(2-((S)-py-
rrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene (47 mg),
(S)-2-(methoxycarbonylamino)propanoic acid (21.7 mg, 0.148 mmol)
and DIEA (0.086 mL, 0.49 mmol) in DMF (2 mL). The reaction was
stirred 5 h at room temperature, diluted with MeOH (2 mL) and water
(2 mL) concentrated to remove the volatiles and purified by
preparative HPLC (acetonitrile/water with 0.1% TFA) and then by
preparative HPLC (acetonitrile/water with 10 mM ammonium acetate)
to afford methyl
((1S)-2-((2S)-2-(5-(7-(2-((2S)-1-(N-(methoxycarbonyl)-L-alanyl)-2-pyrroli-
dinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidi-
nyl)-1-methyl-2-oxoethyl)carbamate (20.9 mg) as white solid. LC-MS
retention time 1.16 min; calcd. for C.sub.42H.sub.44N.sub.8O.sub.6
756.34; found m/z 757.27 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA. .sup.1H NMR (500 MHz,
MeOD) .delta. ppm 8.16-8.24 (m, 2H), 7.87-8.02 (m, 4H), 7.80-7.87
(m, 2H), 7.60-7.75 (m, 4H), 5.42-5.46 (m, 0.3H), 5.34 (dd, J=7.9,
4.0 Hz, 1.7H), 4.55 (q, J=7.0 Hz, 1.7H), 4.31-4.37 (m, 0.3H),
3.80-3.98 (m, 4H), 3.67 (s, 5H), 3.47 (s, 1H), 2.39-2.55 (m, 2H),
2.04-2.36 (m, 4H), 1.92-2.01 (m, 2H), 1.31-1.42 (m, 6H).
##STR00616##
Example 14
Methyl
((1R)-1-(((2S)-2-(5-(7-(2-((2S)-1-((2R)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-ben-
zimidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0918] HATU (56.2 mg, 0.148 mmol) was added to a stirred solution
of a TFA salt of
2-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-5-yl)-7-(2-((S)-py-
rrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene (47 mg),
(R)-2-(methoxycarbonylamino)-3-methylbutanoic acid (25.9 mg, 0.148
mmol) and DIEA (0.086 mL, 0.49 mmol) in DMF (2 mL). The reaction
was stirred 5 h at room temperature, diluted with MeOH (2 mL) and
water (2 mL) concentrated to remove the volatiles and purified by
preparative HPLC (acetonitrile/water with 0.1% TFA) and then by
preparative HPLC (acetonitrile/water with 10 mM ammonium acetate)
to afford a TFA salt of methyl
((1R)-1-(((2S)-2-(5-(7-(2-((2S)-1-((2R)-2-((methoxycarbonyl)amino)-
-3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-be-
nzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
(25 mg) as beige solid. LC-MS retention time 1.36 min; calcd. for
C.sub.46H.sub.52N.sub.8O.sub.6 812.40; found m/z 813.36
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.33 (s,
2H), 8.07-8.13 (m, 4H), 8.05 (dd, J=8.5, 1.2 Hz, 2H), 7.84-7.96 (m,
4H), 5.49 (dd, J=8.6, 3.7 Hz, 2H), 4.36 (d, J=7.3 Hz, 2H),
4.15-4.23 (m, 2H), 3.83-3.97 (m, 2H), 3.72 (s, 6H), 2.58-2.71 (m,
2H), 2.21-2.38 (m, 6H), 2.05-2.18 (m, 2H), 1.07 (d, J=6.7 Hz, 6H),
1.04 (d, J=6.7 Hz, 6H).
##STR00617##
Example 15
5,5'-(2,7-Naphthalenediyl)bis(2-((2S)-1-((2R)-2-phenyl-2-(1-piperidinyl)ac-
etyl)-2-pyrrolidinyl)-1H-benzimidazole)
[0919] HATU (56.2 mg, 0.148 mmol) was added to a stirred solution
of a TFA salt of
2-(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-5-yl)-7-(2-((S)-py-
rrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene (47 mg),
(R)-2-phenyl-2-(piperidin-1-yl)acetic acid (32.4 mg, 0.148 mmol)
and DIEA (0.086 mL, 0.49 mmol) in DMF (2 mL). The reaction was
stirred 5 h at room temperature, diluted with MeOH (2 mL) and water
(2 mL) concentrated to remove the volatiles and purified by
preparative HPLC (acetonitrile/water with 0.1% TFA) and then by
preparative HPLC (acetonitrile/water with 10 mM ammonium acetate)
to afford
5,5'-(2,7-naphthalenediyl)bis(2-((2S)-1-((2R)-2-phenyl-2-(1-piperidinyl)a-
cetyl)-2-pyrrolidinyl)-1H-benzimidazole) (16.4 mg) as white solid.
LC-MS retention time 1.21 min; calcd. for [M+H].sup.+
C.sub.58H.sub.60N.sub.8O.sub.6 900.48; found m/z 451.43
1/2[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.18-8.28 (m, 2H),
7.96-8.05 (m, 2.2H), 7.92 (d, J=1.0 Hz, 1.5H), 7.80-7.90 (m, 2.2H),
7.65-7.76 (m, 3.8H), 7.55-7.64 (m, 3.2H), 7.46-7.55 (m, 4.5H),
6.85-6.97 (m, 1.6H), 6.76-6.83 (m, 1H), 5.65-5.70 (m, 0.5H), 5.28
(dd, J=7.5, 3.3 Hz, 1.5H), 4.48 (br s, 0.5H), 4.02-4.12 (m, 1.5H),
3.90-3.98 (m, 0.5H), 3.74-3.84 (m, 0.5H), 3.25-3.40 (m, 3H),
2.51-2.93 (m, 8H), 1.89-2.33 (m, 8H), 1.62-1.75 (m, 8H), 1.47-1.59
(m 4H).
##STR00618##
Example 16
Methyl
((1S)-1-(((2S)-2-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imi-
dazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0920] HATU (40 mg, 0.11 mmol) was added to a mixture of a TFA salt
of
2-((S)-pyrrolidin-2-yl)-6-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)na-
phthalen-2-yl)-1H-benzo[d]imidazole (32 mg), diisopropylethylamine
(62 .mu.L, 0.35 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (19 mg, 0.106
mmol) in DMF (2 mL) and the resulting mixture was stirred at
ambient conditions for 9 hours. The reaction mixture was diluted
with methanol (2 mL) and water (2 mL). The volatile component was
removed in vacuo and the residue was purified by reverse phase HPLC
(once with water/acetonitrile 10 mM ammonium acetate and twice with
water/acetonitrile 0.1% TFA) to provide a TFA salt of methyl
((1S)-1-(((2S)-2-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol--
2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate (21 mg) as
white solid. LC-MS retention time 1.17 min; calcd. for
C.sub.42H.sub.50N.sub.8O.sub.6 762.39; found m/z 763.34
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.30 (s, 1H), 8.27 (s,
1H), 8.15 (d, J=8.9 Hz, 1H), 8.10 (d, J=8.9 Hz, 1H), 8.07 (s, 1H),
7.94-8.01 (m, 3H), 7.82-7.89 (m, 2H), 5.38 (t, J=7.2 Hz, 1H), 5.31
(t, J=7.6 Hz, 1H), 4.28 (dd, J=14.5, 7.2 Hz, 2H), 4.15 (br s, 2H),
3.86-4.01 (m, 2H), 3.69 (s, 6H), 2.55-2.69 (m, 2H), 2.19-2.39 (m,
6H), 2.04-2.15 (m, 2H), 0.86-1.05 (m, 12H).
##STR00619##
Example 17
Methyl
((1S)-1-(((2S)-2-(4-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imi-
dazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0921] HATU (20 mg, 0.053 mmol) was added to a mixture of a TFA
salt of
2-((S)-pyrrolidin-2-yl)-6-(7-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-4-yl)na-
phthalen-2-yl)-1H-benzo[d]imidazole (16 mg), diisopropylethylamine
(31 .mu.L, 0.18 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (9.3 mg, 0.053
mmol) in DMF (2 mL) and the resulting mixture was stirred at
ambient for 9 hours. The reaction mixture was diluted with methanol
(2 mL) and water (2 mL). The volatile component was removed in
vacuo and the residue was purified by a reverse phase HPLC
(water/acetonitrile 0.1% TFA) to provide a TFA salt of methyl
((1S)-1-(((2S)-2-(4-(7-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol--
2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate (7.9 mg) as
white solid. LC-MS retention time 1.20 min; calcd. for
C.sub.42H.sub.50N.sub.8O.sub.6 762.39; found m/z 763.35
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.35 (s, 1H), 8.25 (s,
1H), 8.09 (dd, J=8.4, 3.2 Hz, 2H), 8.04 (s, 1H), 7.89-8.00 (m, 3H),
7.77-7.87 (m, 2H), 5.37 (t, J=7.0 Hz, 1H), 5.30 (t, J=7.3 Hz, 1H),
4.28 (dd, J=15.4, 7.2 Hz, 2H), 4.08-4.20 (m, 2H), 3.86-4.01 (m,
2H), 3.69 (s, 6H), 2.56-2.68 (m, 3H), 2.39-2.02 (m, 7H), 0.87-1.04
(m, 12H).
##STR00620##
Example 18
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-
-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbo-
nyl)-2-methylpropyl)carbamate
[0922] HATU (218 mg, 0.574 mmol) was added to a mixture of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)naphthalene (100 mg), diisopropylethylamine (334 .mu.L, 1.91
mmol) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (101
mg, 0.574 mmol) in DMF (2 mL) and the resulting mixture was stirred
at ambient for 4 hours. The reaction mixture was diluted with
methanol (2 mL) and water (2 mL). Then, the volatile component was
removed in vacuo and the residue was purified by a reverse phase
HPLC (water/acetonitrile 0.1% TFA) to provide a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate (85 mg) as yellow solid. LC-MS retention
time 1.33 min; calcd. for C.sub.48H.sub.52N.sub.8O.sub.6: 836.4;
found m/z 837.32 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.22 (s, 2H), 8.05-8.13 (m, 4H),
7.96-8.02 (m, 2H), 7.82-7.92 (m, 4H), 5.22-5.32 (m, 2H), 4.59 (d,
J=6.4 Hz, 2H), 3.86 (t, J=6.0 Hz, 2H), 3.60-3.75 (m, 6H), 2.66-2.79
(m, 2H), 2.54 (m, 2H), 2.16-2.26 (m, 2H), 2.02-2.16 (m, 2H),
1.10-1.16 (m, 2H), 0.98-1.05 (m, 6H), 0.94-0.97 (m, 2H), 0.91 (d,
J=6.7 Hz, 6H).
##STR00621##
Example 19
(1R)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2R)-2-(Diethylamino)-2-pheny-
lacetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-2-naphthyl)-1H-
-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-N,N-diethyl-2-oxo-1-pheny-
lethanamine
[0923] HATU (109 mg, 0.287 mmol) was added to a mixture of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)naphthalene (50 mg), diisopropylethylamine (0.167 mL, 0.957
mmol) and (R)-2-(diethylamino)-2-phenylacetic acid, HCl (70.0 mg,
0.287 mmol) in DMF (2 mL) and the resulting mixture was stirred at
ambient for 5 hours. The reaction mixture was diluted with methanol
(2 mL) and water (2 mL). Then, the volatile component was removed
in vacuo and the residue was purified by a reverse phase HPLC
(water/acetonitrile 0.1% TFA) then by reverse phase HPLC
(water/acetonitrile 10 mM ammonium acetate) and finally by reverse
phase HPLC (water/acetonitrile 0.1% TFA) to provide a TFA salt of
(1R)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2R)-2-(diethylamino)-2-phen-
ylacetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-6-yl)-2-naphthyl)-1-
H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-N,N-diethyl-2-oxo-1-phen-
ylethanamine (25.2 mg) as white powder. LC-MS retention time 1.15
min; calcd. for [M+H].sup.+ C.sub.58H.sub.60N.sub.8O.sub.2: 900.48;
found m/z 451.46 [1/2 M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.27 (s, 2H),
8.08-8.17 (m, 4H), 7.96 (td, J=8.7, 1.5 Hz, 4H), 7.87 (d, J=8.6 Hz,
2H), 7.68-7.72 (m, 4H), 7.57-7.68 (m, 6H), 5.79 (s, 2H), 5.23 (dd,
J=9.0, 6.3 Hz, 2H), 3.95-4.05 (m, 2H), 3.25-3.48 (m, 4H), 3.06 (br
s, 4H), 2.43-2.61 (m, 4H), 2.01-2.11 (m, 2H), 1.31 (t, J=6.9 Hz,
12H), 0.62 (ddd, J=8.7, 6.0, 5.8 Hz, 2H), -0.21 (td, J=5.7, 2.4 Hz,
2H).
##STR00622##
Example 20
Methyl
((1R)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-2-phenylacetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5--
yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-
-phenylethyl)carbamate and
Example 21
Dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-aza-
bicyclo[3.1.0]hexane-3,2-diyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarb-
amate
[0924] HATU (109 mg, 0.287 mmol) was added to a mixture of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)naphthalene (50 mg), diisopropylethylamine (0.167 mL, 0.957
mmol) and (R)-2-(methoxycarbonylamino)-2-phenylacetic acid (60.0
mg, 0.287 mmol) in DMF (2 mL) and the resulting mixture was stirred
at ambient for 5 hours. The reaction mixture was diluted with
methanol (2 mL) and water (2 mL). Then, the volatile component was
removed in vacuo and the residue was purified by a reverse phase
HPLC (water/acetonitrile 0.1% TFA) then by reverse phase HPLC
(water/acetonitrile 10 mM ammonium acetate) and finally by reverse
phase HPLC (water/acetonitrile 0.1% TFA) to provide a TFA salt of
methyl
((1R)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-phenylacetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-2--
naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-pheny-
lethyl)carbamate (6.9 mg) as white solid and a TFA salt of dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabicyclo[-
3.1.0]hexane-3,2-diyl((1R)-2-oxo-1-phenyl-2,1-ethanediyl)))biscarbamate
(12.5 mg) as white solid.
[0925] Analytical data for Example 20: LC-MS retention time 1.46
min; calcd. for C.sub.54H.sub.48N.sub.8O.sub.6: 904.37; found m/z
453.35 [1/2 M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.28 (d, J=4.6 Hz, 2H), 8.15
(d, J=8.6 Hz, 2H), 8.12 (s, 1H), 8.07 (s, 1H), 8.03 (t, J=8.9 Hz,
2H), 7.92-7.98 (m, 2H), 7.90 (d, J=8.6 Hz, 1H), 7.85 (d, J=8.6 Hz,
1H), 7.48-7.54 (m, 2H), 7.37-7.48 (m, 8H), 5.85 (s, 1H), 5.78 (s,
1H), 5.39 (dd, J=9.0, 6.3 Hz, 1H), 5.29 (dd, J=9.2, 5.8 Hz, 1H),
3.91-3.97 (m, 1H), 3.68 (s, 3H), 3.66 (s, 3H), 3.52-3.61 (m, 1H),
2.73 (dd, J=13.7, 9.2 Hz, 1H), 2.46-2.65 (m, 3H), 2.06-2.14 (m,
1H), 1.98-2.06 (m, 1H), 1.23-1.30 (m, 1H), 1.10 (br s, 1H),
0.66-0.72 (m, 1H), 0.01-0.10 (m, 1H).
[0926] Analytical data for Example 21: LC-MS retention time 1.48
min; calcd. for [M+H].sup.+ C.sub.54H.sub.48N.sub.8O.sub.6: 904.37;
found m/z 453.35 [1/2 M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.29 (s, 2H),
8.16 (d, J=8.6 Hz, 2H), 8.11 (s, 2H), 8.01-8.06 (m, 2H), 7.95 (d,
J=8.6 Hz, 2H), 7.90 (m, J=8.6 Hz, 2H), 7.48-7.55 (m, 4H), 7.36-7.48
(m, 6H), 5.85 (s, 2H), 5.29 (dd, J=9.2, 5.8 Hz, 2H), 3.92-3.97 (m,
2H), 3.66 (s, 6H), 2.55-2.66 (m, 2H), 2.51 (ddd, J=13.3, 6.6, 6.4
Hz, 2H), 1.97-2.07 (m, 2H), 0.63-0.74 (m, 2H), -0.02-0.10 (m,
2H).
##STR00623##
Example 22
Methyl
((1S)-1-(((2S)-2-(6-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyridin-6-yl)-2-naphthy-
l)-1H-imidazo[4,5-b]pyridin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)-
carbamate and
Example 23
Methyl
((1S)-1-(((2R)-2-(6-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyridin-6-yl)-2-naphthy-
l)-1H-imidazo[4,5-b]pyridin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)-
carbamate
[0927] HATU (63.5 mg, 0.167 mmol) was added to a stirred slurry of
a hydrochloride salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-3H-imidazo[4,5-b]pyridin-6-yl)naphthalene
(45 mg) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid
(29.3 mg, 0.167 mmol) in DMF (1 mL) and DIPEA (0.122 mL, 0.696
mmol). The reaction mixture was stirred at room temperature for 16
h, diluted with MeOH, filtered and purified by preparative HPLC
(acetonitrile/water with 10 mM ammonium acetate) to provide methyl
((1S)-1-(((2S)-2-(6-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyridin-6-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyridin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate (19.2 mg) as white solid and methyl
((1S)-1-(((2R)-2-(6-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyridin-6-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyridin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate (16.5 mg) as white solid (uncertain if pyrrolidine stereocenter
racemized during this step or in a previous step). Resolution of
2,6-bis(2-((S)-pyrrolidin-2-yl)-3H-imidazo[4,5-b]pyridin-6-yl)naphthalene
by chiral HPLC showed a broad peak, but individual diastereomers
could not be resolved under the attempted conditions.
[0928] Analytical data for Example 22: LC-MS retention time 1.407
min; m/z 815.60 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.82-8.71 (m, 2H),
8.32-8.18 (m, 4H), 8.14-8.07 (m, 2H), 7.92-7.84 (m, 2H), 5.37-5.29
(m, 2H), 4.30 (d, J=7.3 Hz, 2H), 4.13-4.04 (m, 2H), 4.02-3.92 (m,
2H), 3.68 (s, 6H), 2.54-1.98 (m, 10H), 1.03-0.89 (m, 12H).
[0929] Analytical data for Example 23: LC-MS retention time 1.442
min; m/z 815.59 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.89-8.66 (m, 2H),
8.36-8.20 (m, 4H), 8.15-8.10 (m, 2H), 7.94-7.86 (m, 2H), 5.44-5.29
(m, 2H), 4.34 (d, J=7.6 Hz, 1H), 4.30 (d, J=7.6 Hz, 1H), 4.21-3.80
(m, 4H), 3.74 (s, 3H), 3.68 (s, 3H), 2.69-1.95 (m, 10H), 1.09-0.41
(m, 12H).
##STR00624##
Example 24
Methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthy-
l)-1H-imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)-
carbamate and
Example 25A
Methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2R)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthy-
l)-1H-imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)-
carbamate and
Example 25B
Methyl
((1S)-1-(((2R)-2-(5-(6-(2-((2R)-1-((2S)-2-((methoxycarbonyl)amino)--
3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthy-
l)-1H-imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)-
carbamate
[0930] HATU (63 mg, 0.17 mmol) was added to a stirred slurry of an
HCl salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-imidazo[4,5-b]pyrazin-5-yl)nap-
hthalene (45 mg) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic
acid (29 mg, 0.17 mmol) in DMF (1 mL) and DIPEA (0.12 mL, 0.69
mmol). The reaction mixture was stirred at room temperature for 16
h diluted with MeOH, filtered and purified by preparative HPLC
(acetonitrile/water with 10 mM ammonium acetate) to yield methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate (11.2 mg) as yellow solid, methyl
((1S)-1-(((2S)-2-(5-(6-(2-((2R)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate (17.3 mg) as yellow solid and methyl
((1S)-1-(((2R)-2-(5-(6-(2-((2R)-1-((2S)-2-((methoxycarbonyl)amino)-3-meth-
ylbutanoyl)-2-pyrrolidinyl)-1H-imidazo[4,5-b]pyrazin-5-yl)-2-naphthyl)-1H--
imidazo[4,5-b]pyrazin-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbam-
ate (6.7 mg) as yellow solid (uncertain if pyrrolidine stereocenter
racemized during this step or in a previous step). Resolution of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-imidazo[4,5-b]pyrazin-5-yl)naphthalene
by chiral HPLC showed a broad peak, but individual diastereomers
could not be resolved under the attempted conditions.
[0931] Analytical data for Example 24: LC-MS retention time 1.420
min; m/z 817.59 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 9.14-9.06 (m, 2H),
8.71-8.64 (m, 2H), 8.37-8.29 (m, 2H), 8.18-8.11 (m, 2H), 5.59-5.30
(m, 2H), 4.30 (d, J=7.3 Hz, 2H), 4.13-3.96 (m, 4H), 3.78-3.64 (m,
6H), 2.57-2.01 (m, 10H), 1.07-0.92 (m, 12H).
[0932] Analytical data for Example 25A: LC-MS retention time 1.507
min; m/z 817.56 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 9.14-9.02 (m, 2H),
8.69-8.60 (m, 2H), 8.36-8.27 (m, 2H), 8.15-8.08 (m, 2H), 5.41-5.36
(m, 1H), 5.34-5.29 (m, 1H), 4.35 (d, J=7.9 Hz, 1H), 4.31 (d, J=7.3
Hz, 1H), 4.24-3.81 (m, 4H), 3.78 (s, 3H), 3.68 (s, 3H), 2.62-1.97
(m, 10H), 1.09-0.46 (m, 12H).
[0933] Analytical data for Example 25B: LC-MS retention time 1.562
min; m/z 817.58 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
acetonitrile/95% water/10 mM ammonium acetate and Solvent B was 5%
water/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00625##
Example 26
Dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrroli-
dinediyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanediyl)))biscarba-
mate
[0934] HATU (114 mg, 0.301 mmol) was added to a stirred solution of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)naphthalene
(50 mg), diisopropylethylamine (175 .mu.L, 1.0 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (65 mg, 0.30 mmol) in DMF (5 mL) and the resulting mixture was
stirred at ambient conditions for 2 hours. The reaction mixture was
diluted with methanol (2 mL) and water (2 mL). Then, the volatile
component was removed in vacuo and the residue was purified by a
reverse phase HPLC (water/acetonitrile/TFA) to provide a TFA salt
of dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(2S)-2,1-pyrrolidinediyl-
((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanediyl)))biscarbamate
(51.5 mg) as pale yellow solid. LC-MS retention time 1.117 min; m/z
897.31 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt 500 MHz, MeOD) .delta.
ppm 8.27 (br s, 2H), 8.10-8.18 (m, 4H), 8.01-8.07 (m, 2H),
7.86-7.96 (m, 4H), 5.88-5.93 (m, 0.2H), 5.39 (t, J=7.0 Hz, 1.8H),
4.33-4.38 (m, 2H), 4.12-4.22 (m, 2H), 3.89-4.05 (m, 7H), 3.70 (s,
6H), 3.35-3.45 (m, 2H), 2.60-2.75 (m, 2H), 2.14-2.47 (m, 6.5H),
1.94-2.12 (m, 2.5H), 1.32-1.69 (m, 8H).
##STR00626##
Example 27
Methyl
((1S)-2-((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-
-methylbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benz-
imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)car-
bamate
[0935] HATU (87 mg, 0.23 mmol) was added to a stirred solution of
methyl
((1S)-2-methyl-1-(((2S)-2-(5-(6-(2-((2S)-2-pyrrolidinyl)-1H-benzimidazol--
5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-1-pyrrolidinyl)carbonyl)propyl)car-
bamate (50 mg), diisopropylethylamine (133 .mu.L, 0.76 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (50 mg, 0.23 mmol) in DMF (3 mL) and the resulting mixture was
stirred at ambient conditions for 2 hours. The reaction mixture was
diluted with methanol (2 mL) and water (2 mL). Then the volatile
component was removed in vacuo and the residue was purified by a
reverse phase HPLC (water/acetonitrile/TFA) to provide a TFA salt
of methyl
((1S)-2-((2S)-2-(5-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-methy-
lbutanoyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-benzimidaz-
ol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
(36 mg) as yellow solid. LC-MS retention time 1.177 min; m/z 855.30
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.22-8.30 (m, 1.8H), 8.20 (br s, 0.2H), 8.07-8.17 (m, 4H),
8.00-8.06 (m, 1.8H), 7.98 (br s, 0.2H), 7.85-7.95 (m, 4H), 5.86 (m,
0.1H), 5.35-5.44 (m, 1.8H), 5.25-5.31 (m, 0.1H), 4.36 (d, J=7.6 Hz,
0.5H), 4.31 (d, J=6.7 Hz, 1H), 4.15 (br s, 1.5H), 3.87-4.05 (m,
4H), 3.69 (br s, 6H), 3.34-3.47 (m, 1H), 2.96-3.23 (m, 2H),
2.61-2.77 (m, 2.5H), 2.21-2.47 (m, 6H), 1.95-2.16 (m, 2H),
1.33-1.69 (m, 3.5H), 1.00-1.05 (m, 0.3H), 0.94-0.99 (m, 3H), 0.91
(d, J=6.7 Hz, 2.7H).
##STR00627##
Example 28
Methyl
((1S)-2-((2S)-2-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-
-(tetrahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-
-naphthyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran--
4-yl)ethyl)carbamate
[0936] HATU (36.6 mg, 0.096 mmol) was added to a stirred solution
of a TFA salt of
2-((S)-pyrrolidin-2-yl)-6-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-
-5-yl)naphthalen-2-yl)-1H-benzo[d]imidazole (29 mg),
diisopropylethylamine (56 .mu.L, 0.32 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (21 mg, 0.10 mmol) in DMF (2 mL) and the resulting mixture was
stirred at ambient conditions for 2 hours. The reaction mixture was
diluted with methanol (2 mL) and water (2 mL). Then the volatile
component was removed in vacuo and the residue was purified by a
reverse phase HPLC (water/acetonitrile/TFA) to provide a TFA salt
of methyl
((1S)-2-((2S)-2-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-(tetr-
ahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-benzimidazol-5-yl)-2-napht-
hyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)e-
thyl)carbamate (16.3 mg) as yellow solid. LC-MS retention time
1.023 min; m/z 847.31 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min and an analysis time of 4 min where Solvent A was 10%
acetonitrile/90% water/0.1% TFA and Solvent B was 90%
acetonitrile/10% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.29 (d, J=12.5 Hz, 2H), 8.15
(d, J=8.6 Hz, 1H), 8.10 (d, J=8.5 Hz, 1H), 8.07 (s, 1H), 7.92-8.01
(m, 3H), 7.81-7.90 (m, 2H), 5.37 (dd, J=7.9, 6.1 Hz, 1H), 5.30 (t,
J=7.5 Hz, 1H), 4.28-4.39 (m, 2H), 4.12-4.22 (m, 2H), 3.88-4.04 (m,
7H), 3.65-3.77 (m, 6H), 3.38-3.45 (m, 1H), 2.56-2.69 (m, 2H),
2.20-2.40 (m, 6.5H), 2.01 (br s, 2.5H), 1.57-1.69 (m, 2.5H),
1.35-1.53 (m, 6.5H).
##STR00628##
Example 29
Methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)--
1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
[0937] HATU (57.6 mg, 0.152 mmol) was added to a stirred solution
of a TFA salt of
2-((S)-pyrrolidin-2-yl)-5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-imida-
zol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazole (47 mg),
diisopropylethylamine (88 .mu.L, 0.51 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (26.5 mg, 0.152
mmol) in DMF (4 mL) and the resulting mixture was stirred at
ambient conditions for 2 hours. The reaction mixture was diluted
with methanol (2 mL) and water (2 mL). The volatile component was
removed in vacuo and the residue was purified by a reverse phase
HPLC (water/methanol/TFA), repurified by a reverse phase HPLC
(water/acetonitrile/ammonium acetate) and finally repurified by a
reverse phase HPLC (water/methanol/TFA) to provide a TFA salt of
methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imi-
dazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate (33.3
mg) as yellow solid. LC-MS retention time 1.237 min; m/z 789.29
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min and an
analysis time of 4 min where Solvent A was 10% acetonitrile/90%
water/0.1% TFA and Solvent B was 90% acetonitrile/10% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.29 (d, J=8.9 Hz, 2H), 8.15 (d, J=8.5 Hz, 1H), 8.10 (d, J=8.9
Hz, 1H), 7.95-8.03 (m, 4H), 7.93 (s, 1H), 7.84-7.91 (m, 3H),
5.69-5.72 (m, 0.2H), 5.24-5.35 (m, 1.8H), 4.27 (dd, J=7.2, 2.6 Hz,
2H), 4.09-4.19 (m, 2H), 3.85-3.97 (m, 2H), 3.67-3.76 (m, 6H),
2.55-2.68 (m, 2H), 2.27-2.38 (m, 2H), 2.14-2.27 (m, 4H), 2.02-2.14
(m, 2H), 0.86-1.06 (m, 12H).
##STR00629##
Example 30
Dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-aza-
bicyclo[3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1--
ethanediyl)))biscarbamate
[0938] HATU (54.6 mg, 0.144 mmol) was added to a stirred solution
of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)naphthalene (25 mg), diisopropylethylamine (84 .mu.L, 0.48
mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (31.2 mg, 0.144 mmol) in DMF (5 mL) and the resulting mixture
was stirred at ambient conditions for 4 hours. The reaction mixture
was diluted with methanol (2 mL) and water (2 mL). The volatile
component was removed in vacuo and the residue was purified by a
reverse phase HPLC (water/methanol/TFA), repurified by a reverse
phase HPLC (water/acetonitrile/ammonium acetate) and finally
repurified by a reverse phase HPLC (water/methanol/TFA) to provide
a TFA salt of dimethyl
(2,6-naphthalenediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabicyclo[-
3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanedi-
yl)))biscarbamate (8.2 mg) as off-white solid. LC-MS retention time
1.220 min; m/z 461.3 [1/2 M+H.sup.+]. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.50 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 3 min, a hold time of 1 min and an analysis time of 4 min
where Solvent A was 10% acetonitrile/90% water/0.1% TFA and Solvent
B was 90% acetonitrile/10% water/0.1% TFA. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.28 (br s, 2H),
8.07-8.18 (m, 4H), 8.00-8.07 (m, 2H), 7.84-7.96 (m, 4H), 5.23-5.31
(m, 2H), 4.59-4.67 (m, 2H), 3.85-4.01 (m, 6H), 3.70 (s, 6H),
3.28-3.44 (m, 4H), 2.71-2.80 (m, 2H), 2.58 (ddd, J=13.2, 6.5, 6.3
Hz, 2H), 2.03-2.20 (m, 4H), 1.41-1.65 (m, 8H), 1.13 (br s, 2H),
0.97 (br s, 2H).
##STR00630##
Example 31
Methyl
((1S)-1-(((2S)-2-(4-((6-((2-((2S)-1-((2S)-2-((methoxycarbonyl)amino-
)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)e-
thynyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamat-
e
[0939] HATU (47.6 mg, 0.125 mmol) was added to a TFA salt of
2,6-bis((2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)ethynyl)naphthalene
(38 mg) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (22
mg, 0.13 mmol) in DMF (0.7 mL) and TEA (0.047 mL, 0.33 mmol) and
the clear yellow solution was at rt ON. The reaction was diluted
with MeOH (1 mL), filtered and purified by prep HPLC 15-100%
MeOH/water/TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((2S)-2-(4-((6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-met-
hylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)ethynyl)-
-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
(37.1 mg) as light yellow solid. LC-MS retention time 2.530 min;
m/z 759.61 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00631##
Example 32
Methyl
((1S)-1-(((2S)-2-(4-((6-((2-((2S)-1-((2S)-2-((methoxycarbonyl)amino-
)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)e-
thynyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamat-
e
[0940] A TFA salt of
4,4'-(2,6-naphthalenediyl)bis(2-((2S)-2-pyrrolidinyl)-1H-imidazole)
(21 mg, 0.025 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (12.9 mg) were
dissolved into DMF (3 mL) and DIEA (0.043 mL, 0.246 mmol). Then
HATU (28.0 mg, 0.074 mmol) was added and the reaction mixture was
stirred at room temperature for 2 h. The volatiles were removed
under vacuum and the crude product was purified by prep HPLC
(Waters Sunfire C18 column 30.times.100 mm 5u eluted with a
gradient of 0 to 90% MeOH-Water+0.1% TFA) to yield a TFA salt of
methyl
((1S)-1-(((2S)-2-(4-((6-((2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-me-
thylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)ethynyl-
)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
(10.4 mg) as off white solid. LC-MS retention time 1.158 min; m/z
713.60 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 5 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.31 (s, 2H), 8.11 (d, J=8.6 Hz, 2H), 7.94-8.03 (m, 2H),
7.84-7.94 (m, 2H), 5.29 (t, J=7.3 Hz, 2H), 4.26 (d, J=7.02 Hz, 2H),
4.14 (t, J=10.2 Hz, 2H), 3.85-3.96 (m, 2H), 3.68 (s, 6H), 2.53-2.67
(m, 2H), 2.31 (d, J=10.4 Hz, 2H), 2.16-2.28 (m, 4H), 2.03-2.15 (m,
2H), 0.88-1.08 (m, 12H).
##STR00632##
Example 33
Methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2--
naphthyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H--
pyran-4-yl)ethyl)carbamate
[0941] HATU (120 mg, 0.316 mmol) was added to a stirred solution of
a TFA salt of
2-((2S)-2-pyrrolidinyl)-4-(4-(6-(2-((2S)-2-pyrrolidinyl)-1H-imida-
zol-4-yl)-2-naphthyl)phenyl)-1H-imidazole (50 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (68.7 mg, 0.316 mmol) in DMF (5 mL) and DIEA (0.18 mL, 1.1
mmol) and the reaction mixture was stirred at room temperature for
2 h. The reaction mixture was diluted with MeOH (2 mL) and water (2
mL) and the volatiles were removed under vacuum. The residue was
purified by prep HPLC (Waters Sunfire C18 column 30.times.100 mm 5u
eluted with a gradient of 10 to 90% MeOH-Water+0.1% TFA) and then
repurified by prep HPLC (Waters Sunfire C18 column 30.times.150 mm
5u eluted with a gradient of to % ACN-Water+0.1% TFA) to yield a
TFA salt of methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-(t-
etrahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphth-
yl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro-2H-pyran--
4-yl)ethyl)carbamate (96 mg) as yellow solid. LC-MS retention time
1.207 min; m/z 873.63 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Waters Sunfire 5u C18
4.6.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 5 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90%
MeOH/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (TFA salt, 500
MHz, MeOD) .delta. ppm 8.29 (d, J=7.9 Hz, 2H), 8.14 (d, J=8.9 Hz,
1H), 8.06-8.11 (m, 1H), 7.94-8.03 (m, 4H), 7.90-7.94 (m, 1H),
7.80-7.90 (m, 3H), 5.74 (br s, 0.2H), 5.23-5.35 (m, 1.8H), 4.32
(dd, J=8.1, 4.7 Hz, 2H), 4.16 (d, J=2.8 Hz, 2H), 3.94 (dd, J=9.5,
7.3 Hz, 6H), 3.69 (s, 6H), 3.31-3.48 (m, 4H), 2.55-2.68 (m, 2H),
2.16-2.35 (m, 6H), 1.92-2.10 (m, 2H), 1.63 (d, J=12.8 Hz, 2H),
1.34-1.54 (m, 6H).
##STR00633##
Example 34
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-
-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0942] HATU (57 mg, 0.150 mmol) was added to a stirred solution of
(1R,3S,5R)-3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-im-
idazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
exane (25 mg, 0.050 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (33 mg) in DMF (3 mL) and DIEA (0.078 mL, 0.50 mmol) and the
reaction mixture was stirred at room temperature for 2 h. The
reaction mixture was diluted with MeOH (2 mL) and water (2 mL) and
the volatiles were removed under vacuum. The residue was purified
by prep HPLC (Waters Sunfire C18 column 30.times.100 mm 5u eluted
with a gradient of 10 to 100% MeOH-Water+0.1% TFA) to yield a TFA
salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (27 mg)
as pale yellow solid. LC-MS retention time 1.280 min; m/z 897.65
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.28-8.32 (m, 2H), 8.14 (d, J=8.6 Hz, 1H), 8.07-8.11 (m, 1H),
7.95-8.02 (m, 4H), 7.84-7.93 (m, 4H), 5.12-5.23 (m, 2H), 4.62 (dd,
J=7.6, 4.0 Hz, 2H), 3.92-4.00 (m, 4H), 3.83-3.89 (m, 2H), 3.72-3.78
(m, 2H), 3.70 (s, 6H), 3.35-3.46 (masked with methanol, m, 4H),
2.68-2.77 (m, 2H), 2.47-2.57 (m, 2H), 2.00-2.15 (m, 4H), 1.42-1.65
(m, 6H), 1.06-1.17 (m, 2H), 0.84-0.93 (m, 2H).
##STR00634##
Example 35
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)car-
bonyl)-2-methylpropyl)carbamate
[0943] HATU (57 mg, 0.150 mmol) was added to a stirred solution of
(1R,3S,5R)-3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-im-
idazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
exane (25 mg) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic
acid (26 mg, 0.15 mmol) in DMF (3 mL) and DIEA (0.070 mL, 0.40
mmol) and the reaction mixture was stirred at room temperature for
2 h. The reaction mixture was diluted with MeOH (2 mL) and water (2
mL) and the volatiles were removed under vacuum. The residue was
purified by prep HPLC (Waters Sunfire C18 column 30.times.100 mm 5u
eluted with a gradient of 10 to 100% MeOH-Water+0.1% TFA) to yield
a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)--
2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate (34.8 mg) as pale yellow solid. LC-MS
retention time 1.402 min; m/z 813.69 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.30 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and Solvent B
was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.29 (d, J=7.9 Hz, 2H), 8.14
(d, J=8.9 Hz, 1H), 8.09 (d, J=8.9 Hz, 1H), 7.95-8.01 (m, 4H), 7.92
(s, 1H), 7.84-7.90 (m, 3H), 5.16 (ddd, J=11.6, 9.2, 7.0 Hz, 2H),
4.58 (dd, J=6.4, 2.1 Hz, 2H), 3.81-3.88 (m, 2H), 3.66-3.75 (m, 6H),
2.67-2.77 (m, 2H), 2.46-2.57 (m, 2H), 2.21 (dq, J=12.9, 6.5 Hz,
2H), 2.06-2.14 (m, 2H), 1.08-1.18 (m, 2H), 1.04 (d, J=6.7 Hz, 6H),
0.95 (d, J=6.7 Hz, 6H), 0.91 (br s, 2H).
##STR00635##
Example 36
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-
-5-yl)-2-quinolinyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)car-
bonyl)-2-methylpropyl)carbamate
[0944] HATU (32.7 mg, 0.086 mmol) was added to a solution of a
hydrochloride salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)quinoline (25 mg) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (15 mg, 0.086
mmol) in DMF (1 mL) and DIPEA (0.059 mL, 0.34 mmol) and the mixture
was stirred at rt for 16 h. The reaction was diluted with MeOH, and
purified by prep HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield
a TFA salt of dimethyl
(2S,2'S)-1,1'-((1R,1'R,3S,3'S,5R,5'R)-3,3'-(6,6'-(quinoline-2,6-diyl)bis(-
1H-benzo[d]imidazole-6,2-diyl))bis(2-azabicyclo[3.1.0]hexane-3,2-diyl))bis-
(3-methyl-1-oxobutane-2,1-diyl)dicarbamate (8.3 mg) as a yellow
solid. LC-MS retention time 1.88 min; m/z 838 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna3.0.times.50 mm S10. Solvent A=90% water:10%
methanol: 0.1% TFA. Solvent B=10% water:90% methanol: 0.1% TFA.
Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient Time=3
min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.64-8.80 (1H, m), 8.49-8.62 (1H, m), 7.45-8.48 (9H, m), 5.21-5.52
(2H, m), 4.50-4.68 (2H, m), 3.82-4.13 (2H, m), 3.70-3.79 (2H, m),
3.68, 3.66 (6H, s, s), 2.48-2.83 (3H, m), 2.02-2.28 (3H, m),
1.09-1.30 (2H, m), 0.83-1.08 (14H, m).
##STR00636##
Example 37
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-
-5-yl)-2-quinoxalinyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)c-
arbonyl)-2-methylpropyl)carbamate
[0945] HATU (25 mg, 0.066 mmol) was added to a solution of a TFA
salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)quinoxaline (28 mg) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (11.5 mg, 0.066
mmol) in DMF (0.5 mL) and DIPEA (0.045 mL, 0.26 mmol) and the
mixture was stirred at rt for 2 h. The reaction was diluted with
MeOH, and purified by prep HPLC (H.sub.2O-MeOH with 0.1% TFA
buffer) to yield a TFA salt of dimethyl
(2S,2'S)-1,1'-((1R,1'R,3S,3'S,5R,5'R)-3,3'-(6,6'-(quinoxaline-2,6-diyl)bi-
s(1H-benzo[d]imidazole-6,2-diyl))bis(2-azabicyclo[3.1.0]hexane-3,2-diyl))b-
is(3-methyl-1-oxobutane-2,1-diyl)dicarbamate (27 mg) as a yellow
solid. LC-MS retention time 1.94 min; m/z 839 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna3.0.times.50 mm S10. Solvent A=90% water:10%
methanol: 0.1% TFA. Solvent B=10% water:90% methanol: 0.1% TFA.
Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient Time=3
min. Wavelength=220). .sup.1H NMR (TFA salt, 400 MHz, MeOD) .delta.
ppm 9.56 (s, 1H), 8.67 (s, 1H), 8.52 (d, J=8.8 Hz, 1H), 8.40 (s,
1H), 8.26 (d, J=8.8 Hz, 1H), 8.24 (d, J=8.8 Hz, 1H), 8.17 (s, 1H),
8.05 (d, J=8.5 Hz, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.90 (d, J=8.8 Hz,
1H), 5.30 (app dd, J=9.0, 6.8 Hz, 2H), 4.60 (app d, J=6.0 Hz, 2H),
3.93-3.84 (m, 2H), 3.68 (s, 6H), 2.80-2.70 (m, 2H), 2.63-2.51 (m,
2H), 2.29-2.09 (m, 4H), 1.21-1.11 (m, 2H), 1.03 (d, J=6.8 Hz, 6H),
1.00-0.90 (m, 2H), 0.93 (d, J=6.5 Hz, 6H).
##STR00637##
Example 38
[0946] Dimethyl
(2,6-quinoxalinediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabicyclo[-
3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-ethanedi-
yl)))biscarbamate
[0947] HATU (25.9 mg, 0.068 mmol) was added to a solution of a TFA
salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)quinoxaline (29 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (14.8 mg, 0.068 mmol) in DMF (0.5 mL) and DIPEA (0.046 mL,
0.27 mmol) and the mixture was stirred at rt for 16 h. The reaction
was diluted with MeOH, and purified by prep HPLC (H.sub.2O-MeOH
with 0.1% TFA buffer) to yield a TFA salt of dimethyl
(1S,1'S)-2,2'-((1R,1'R,3S,3'S,5R,5'R)-3,3'-(6,6'-(quinoxaline-2,6-diyl)bi-
s(1H-benzo[d]imidazole-6,2-diyl))bis(2-azabicyclo[3.1.0]hexane-3,2-diyl))b-
is(2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethane-2,1-diyl)dicarbamate
(32.5 mg) as a yellow solid. LC-MS retention time 1.74 min; m/z 462
[1/2 M+H].sup.+. (Column PHENOMENEX.RTM. Luna3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220). .sup.1H NMR (TFA salt, 400
MHz, MeOD) .delta. ppm 9.53 (s, 1H), 8.70 (s, 1H), 8.55 (dd, J=8.8,
1.5 Hz, 1H), 8.44 (d, J=1.8 Hz, 1H), 8.31 (d, J=8.8 Hz, 1H), 8.27
(dd, J=8.8, 1.8 Hz, 1H), 8.19 (br s, 1H), 8.08 (dd, J=8.8, 1.5 Hz,
1H), 7.96 (d, J=8.8 Hz, 1H), 7.92 (d, J=8.8 Hz, 1H), 5.28 (app dd,
J=8.8, 6.8 Hz, 2H), 4.63 (app d, J=7.3 Hz, 2H), 3.99-3.86 (m, 6H),
3.69 (s, 6H), 3.44-3.29 (m, 4H), 2.81-2.71 (m, 2H), 2.63-2.52 (m,
2H), 2.20-2.03 (m, 4H), 1.68-1.37 (m, 8H), 1.17-1.08 (m, 2H),
0.99-0.92 (m, 2H).
##STR00638##
Example 39
Dimethyl
(2,6-quinolinediylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R)-2-azabi-
cyclo[3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,1-et-
hanediyl)))biscarbamate
[0948] HATU (34.0 mg, 0.089 mmol) was added to a solution of a
hydrochloride salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)quinoline (26 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (19.4 mg, 0.089 mmol) in DMF (0.5 mL) and DIPEA (0.061 mL,
0.350 mmol) and the mixture was stirred at rt for 16 h. The
reaction was diluted with MeOH and purified by prep HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield a TFA salt of
dimethyl
(1S,1'S)-2,2'-((1R,1'R,3S,3'S,5R,5'R)-3,3'-(6,6'-(quinoline-2,6-diyl)bis(-
1H-benzo[d]imidazole-6,2-diyl))bis(2-azabicyclo[3.1.0]hexane-3,2-diyl))bis-
(2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethane-2,1-diyl)dicarbamate (6.0
mg) as a yellow solid. LC-MS retention time 1.65 min; m/z 462 [1/2
M+H].sup.+. (Column PHENOMENEX.RTM. Luna3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.70 (1H, d, J=8.5 Hz), 8.57 (1H, s), 8.23-8.45 (5H,
m), 8.17 (1H, s), 8.01-8.12 (1H, m), 7.88-7.97 (2H, m), 5.27 (2H,
t, J=7.8 Hz), 4.33-4.68 (2H, m), 3.84-4.02 (6H, m), 3.69 (6H, s),
3.35-3.47 (4H, m), 2.67-2.81 (2H, m), 2.49-2.64 (2H, m), 1.91-2.21
(4H, m), 1.35-1.71 (8H, m), 1.07-1.17 (2H, m), 0.85-1.00 (2H,
m).
##STR00639##
Example 40
Methyl
((1S)-2-((2S)-2-(4-chloro-5-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-imidazol-
-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetra-
hydro-2H-pyran-4-yl)ethyl)carbamate
[0949] HATU (45 mg, 0.118 mmol) was added to a solution of
4-chloro-2-((S)-pyrrolidin-2-yl)-5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-imid-
azol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazole (20 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (25.6 mg, 0.118 mmol) in DMF (3 mL) and DIEA (0.070 mL, 0.40
mmol) and the reaction mixture was stirred at room temperature for
2 h. The reaction mixture was diluted with MeOH (2 mL) and water (2
mL) and the volatiles were removed under vacuum. The residue was
purified by prep HPLC (Waters Sunfire C18 column 30.times.100 mm 5u
eluted with a gradient of 10 to 100% MeOH-Water+0.1% TFA) and then
repurified by HPLC (Waters Sunfire C18 column 30.times.150 mm 5u
eluted with a gradient of 10 to 100% ACN-Water+0.1% TFA) to yield a
TFA salt of methyl
((1S)-2-((2S)-2-(4-chloro-5-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-
-2-naphthyl)phenyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)-2-oxo-1-(tetrahydro--
2H-pyran-4-yl)ethyl)carbamate (12.9 mg) as yellow solid. LC-MS
retention time 1.595 min; m/z 907.39 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.30 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and Solvent B
was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.20-8.28 (m, 2H), 8.09-8.12
(m, 1H), 8.02-8.08 (m, 1H), 7.91-8.00 (m, 3H), 7.87-7.91 (m, 2H),
7.80-7.87 (m, 2H), 5.28 (t, J=7.5 Hz, 1H), 5.06-5.14 (m, 1H),
4.25-4.35 (m, 2H), 4.11-4.19 (m, 1H), 4.00-4.07 (m, 1H), 3.85-4.00
(m, 6H), 3.63-3.69 (m, 6H), 3.32-3.43 (m, 4H), 2.57-2.64 (m, 1H),
2.34-2.40 (m, 1H), 2.26-2.32 (m, 2H), 2.12-2.25 (m, 4H), 1.95-2.09
(m, 2H), 1.34-1.64 (m, 8H).
##STR00640##
Example 41
Methyl
((1S)-1-(((2S)-2-(4-(6-(4-(4-chloro-2-((2S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)phenyl)-2-n-
aphthyl)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbama-
te
[0950] HATU (45 mg, 0.118 mmol) was added to a solution of
4-chloro-2-((S)-pyrrolidin-2-yl)-5-(4-(6-(2-((S)-pyrrolidin-2-yl)-1H-imid-
azol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazole (20 mg) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (20.7 mg, 0.118
mmol) in DMF (3 mL) and DIEA (0.070 mL, 0.40 mmol) and the reaction
mixture was stirred at room temperature for 2 h. The reaction
mixture was diluted with MeOH (2 mL) and water (2 mL) and the
volatiles were removed under vacuum. The residue was purified by
prep HPLC (Waters Sunfire C18 column 30.times.100 mm 5u eluted with
a gradient of 10 to 100% MeOH-Water+0.1% TFA) and then repurified
by HPLC (Waters Sunfire C18 column 30.times.150 mm 5u eluted with a
gradient of 10 to 100% ACN-Water+0.1% TFA) to yield a TFA salt of
methyl
((1S)-1-(((2S)-2-(4-(6-(4-(4-chloro-2-((2S)-1-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-5-yl)phenyl)-2-naphthy-
l)-1H-imidazol-2-yl)-1-pyrrolidinyl)carbonyl)-2-methylpropyl)carbamate
(3.6 mg) as yellow solid. LC-MS retention time 1.715 min; m/z
823.49 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.24-8.29 (m, 1H), 8.22 (s, 1H), 8.08-8.15 (m, 1H), 8.03-8.08
(m, 1H), 7.81-8.01 (m, 7H), 5.31 (t, J=7.5 Hz, 1H), 5.12 (app t,
J=7.5 Hz, 1H), 4.21-4.32 (m, 2H), 4.08-4.19 (m, 1H), 3.99-4.08 (m,
1H), 3.85-3.96 (m, 2H), 3.57-3.74 (m, 6H), 2.56-2.67 (m, 1H),
2.35-2.46 (m, 1H), 2.28-2.35 (m, 2H), 2.20-2.26 (m, 2H), 2.05-2.19
(m, 4H), 0.90-1.09 (m, 12H).
##STR00641##
Example 42
Methyl
((1S)-2-((1R,3S,5R)-3-(7-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl-
)-1H-imidazol-4-yl)-2-naphthyl)-1H-naphtho[1,2-d]imidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0951] HATU (103 mg, 0.271 mmol) was added to a solution of
2-((S)-pyrrolidin-2-yl)-7-(6-(2-((S)-pyrrolidin-2-yl)-1H-imidazol-5-yl)na-
phthalen-2-yl)-1H-naphtho[1,2-d]imidazole (45 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (58.8 mg, 0.271 mmol) in DMF (3 mL) and DIEA (0.16 mL, 0.90
mmol) and the reaction mixture was stirred at room temperature for
2 h. The reaction mixture was diluted with MeOH (2 mL) and water (2
mL) and the volatiles were removed under vacuum. The residue was
purified by prep HPLC (Waters Sunfire C18 column 30.times.100 mm 5u
eluted with a gradient of 10 to 100% MeOH-Water+0.1% TFA) and then
repurified by HPLC (Waters Sunfire C18 column 30.times.150 mm 5u
eluted with a gradient of 0 to 90% ACN-Water+0.1% TFA) to yield a
TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(7-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-i-
midazol-4-yl)-2-naphthyl)-1H-naphtho[1,2-d]imidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (8.7
mg) as white solid. LC-MS retention time 1.307 min; m/z 897.67
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.51-8.57 (m, 2H), 8.40 (s, 1H), 8.30 (s, 1H), 8.23 (d, J=10.1
Hz, 1H), 8.18 (d, J=8.6 Hz, 1H), 8.05-8.15 (m, 3H), 8.00 (s, 1H),
7.87 (dd, J=8.6, 1.8 Hz, 1H), 7.79-7.85 (m, 1H), 5.43 (t, J=7.3 Hz,
1H), 5.30 (t, J=7.3 Hz, 1H), 4.28-4.40 (m, 2H), 4.19 (br s, 2H),
3.88-4.07 (m, 6H), 3.70 (d, J=1.5 Hz, 6H), 3.26-3.45 (overlap with
methanol, m, 4H), 2.57-2.71 (m, 2H), 2.30-2.42 (m, 3H), 2.20-2.29
(m, 3H), 1.93-2.10 (m, 2H), 1.58-1.68 (m., 2H), 1.34-1.58 (m,
6H).
##STR00642##
Example 43
Methyl
((1S)-2-((1R,3S,5R)-3-(7-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl-
)-1H-imidazol-4-yl)-2-naphthyl)-1H-naphtho[1,2-d]imidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0952] HATU (24.5 mg, 0.065 mmol) was added to a solution of a TFA
salt of
2,6-bis(2-((S)-pyrrolidin-2-yl)-1H-benzo[d]imidazol-6-yl)quinoline
(30 mg) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid
(11.3 mg, 0.065 mmol) in DMF (1 mL) and DIPEA (0.044 mL, 0.252
mmol) and the mixture was stirred at rt for 2 h. The reaction was
diluted with MeOH and purified by prep HPLC (H.sub.2O-MeOH with
0.1% TFA buffer) to yield a TFA salt of dimethyl
(2S,2'S)-1,1'-((2S,2'S)-2,2'-(6,6'-(quinoline-2,6-diyl)bis(1H-benzo[d]imi-
dazole-6,2-diyl))bis(pyrrolidine-2,1-diyl))bis(3-methyl-1-oxobutane-2,1-di-
yl)dicarbamate (34 mg) as a yellow solid. LC-MS retention time 1.82
min; m/z 814 [M+H].sup.+. (Column PHENOMENEX.RTM. Luna3.0.times.50
mm S10. Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10%
water:90% methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0.
Final % B=100. Gradient Time=3 min. Wavelength=220). .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 8.71 (1H, d, J=8.8 Hz), 8.57 (1H, s),
8.21-8.43 (5H, m), 8.18 (1H, s), 8.04 (1H, dd, J=8.5, 1.5 Hz),
7.88-7.96 (2H, m), 5.40 (2H, t, J=7.03 Hz), 4.31 (2H, d, J=6.8 Hz),
4.09-4.21 (2H, m), 3.91-4.06 (2H, m), 3.68 (6H, s), 2.58-2.73 (2H,
m), 2.20-2.49 (6H, m), 2.02-2.19 (2H, m), 0.96 (6H, dd, J=6.8, 1.8
Hz), 0.90 (6H, dd, J=6.7, 1.6 Hz).
##STR00643##
Example 44
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-y-
l)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate
[0953] HATU (82 mg, 0.22 mmol) was added to a solution of
6-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)napht-
halen-2-yl)-2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidaz-
ole (34 mg), (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid
(37.8 mg, 0.216 mmol) and DIEA (0.13 mL, 0.72 mmol) in DMF (3 mL)
and the reaction was stirred 2 h at room temperature. The reaction
mixture was diluted with MeOH (2 ml) and water (2 mL). The
volatiles were removed under vacuum. The residue was purified by
prep HPLC (Waters Sunfire C18 column 30.times.150 mm 5u eluted with
a gradient of 10 to 80% ACN-Water+0.1% TFA) to afford a TFA salt of
methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-n-
aphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-met-
hylpropyl)carbamate (49.8 mg) as pale yellow solid. LC-MS retention
time 1.357 min; m/z 787.65 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire 5u C18 4.6.times.30 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and Solvent B
was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.29 (d, J=10.7 Hz, 2H),
8.07-8.19 (m, 3H), 7.94-8.05 (m, 3H), 7.84-7.92 (m, 2H), 5.29 (dd,
J=9.2, 6.7 Hz, 1H), 5.18 (dd, J=9.2, 7.0 Hz, 1H), 4.59 (dd, J=12.4,
6.6 Hz, 2H), 3.81-3.91 (m, 2H), 3.69 (s, 6H), 2.67-2.80 (m, 2H),
2.48-2.61 (m, 2H), 2.07-2.27 (m, 4H), 1.09-1.19 (m, 2H), 0.99-1.06
(m, 6H), 0.88-1.00 (m, 8H).
##STR00644##
Example 45
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl-
)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0954] HATU (82 mg, 0.216 mmol) was added to a solution of
6-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)napht-
halen-2-yl)-2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidaz-
ole (34 mg),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (46.9 mg, 0.216 mmol) and DIEA (0.13 mL, 0.72 mmol) in DMF (3
mL) and the reaction was stirred for 2 h at room temperature. The
reaction mixture was diluted with MeOH (2 ml) and water (2 mL). The
volatiles were removed under vacuum. The residue was purified by
prep HPLC (Waters Sunfire C18 column 30.times.150 mm 5u eluted with
a gradient of 10 to 80% ACN-Water+0.1% TFA) to afford a TFA salt of
methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-b-
enzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-y-
l)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (38 mg) as
pale yellow solid. LC-MS retention time 1.240 min; m/z 871.66
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.30 (d, J=6.1 Hz, 2H), 8.08-8.18 (m, 3H), 8.04 (dd, J=8.6, 1.5
Hz, 1H), 7.95-8.01 (m, 2H), 7.85-7.92 (m, 2H), 5.27 (dd, J=9.2, 6.7
Hz, 1H), 5.16 (dd, J=9.2, 7.0 Hz, 1H), 4.59-4.66 (m, 2H), 3.84-4.01
(m, 6H), 3.70 (s, 6H), 3.35-3.45 (m, 4H), 2.69-2.80 (m, 2H),
2.48-2.61 (m, 2H), 2.03-2.20 (m, 4H), 1.57-1.65 (m, 2H), 1.40-1.56
(m, 6H), 1.07-1.17 (m, 2H), 0.97 (br s, 1H), 0.91 (br s, 1H).
##STR00645##
Example 46
[0955] Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6'-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)-
amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl-
)-2,2'-binaphthalen-6-yl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-y-
l)carbonyl)-2-methylpropyl)carbamate
[0956] HATU (22.7 mg, 0.060 mmol) was added to a solution of a TFA
salt of
6,6'-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-
-5-yl)-2,2'-binaphthyl (22 mg),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (10.5 mg, 0.060
mmol) and DIEA (0.035 mL, 0.20 mmol) in DMF (3 mL) and the reaction
was stirred at rt for 2 h. The reaction mixture was diluted with
MeOH (2 ml) and water (2 mL) and the volatiles were removed under
vacuum. The residue was purified by prep HPLC (Waters Sunfire C18
column 30.times.100 mm 5u eluted with a gradient of 10 to 80%
MeOH-Water+0.1% TFA) to afford a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6'-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)-
amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl-
)-2,2'-binaphthalen-6-yl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-y-
l)carbonyl)-2-methylpropyl)carbamate (15.6 mg) as tan solid. LC-MS
retention time 1.765 min; m/z 482.68 (1/2 M+H+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Waters Sunfire 5u C18 4.6.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and Solvent B
was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.36 (s, 2H), 8.27 (s, 2H),
8.12-8.18 (m, 4H), 8.11 (s, 2H), 8.05 (app t, J=8.6 Hz, 4H),
7.91-7.96 (m, 2H), 7.86-7.91 (m, 2H), 5.29 (dd, J=9.2, 6.7 Hz, 2H),
4.61 (d, J=6.4 Hz, 2H), 3.85-3.91 (m, 2H), 3.70 (s, 6H), 2.72-2.80
(m, 2H), 2.53-2.62 (m, 2H), 2.12-2.26 (m, 4H), 1.13-1.21 (m, 2H),
1.04 (d, J=6.7 Hz, 6H), 0.99 (br s, 2H), 0.95 (d, J=6.7 Hz,
6H).
##STR00646##
Example 47
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-
-5-yl)-1,5-naphthyridin-2-yl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)carbonyl)-2-methylpropyl)carbamate
[0957] HATU (8.7 mg, 0.023 mmol) was added to a solution of a TFA
salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)-1,5-naphthyridine (12 mg, 9.93 .mu.mol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (4.2 mg) in DMF
(0.5 mL) and DIPEA (0.017 mL, 0.099 mmol) and the mixture was
stirred at rt for 16 h. The reaction was diluted with MeOH, and
purified by prep HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield
a TFA salt of dimethyl
(2S,2'S)-1,1'-((1R,1'R,3S,3'S,5R,5'R)-3,3'-(6,6'-(1,5-naphthyridine-2,6-d-
iyl)bis(1H-benzo[d]imidazole-6,2-diyl))bis(2-azabicyclo[3.1.0]hexane-3,2-d-
iyl))bis(3-methyl-1-oxobutane-2,1-diyl)dicarbamate (5.6 mg) as
yellow solid. LC-MS retention time 1.91 min; m/z 420 [1/2
M+H].sup.+. (Column PHENOMENEX.RTM. Luna3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.59-8.67 (4H, m), 8.48 (2H, dd, J=8.8, 1.3 Hz), 8.44
(2H, d, J=8.8 Hz), 7.92 (2H, d, J=8.5 Hz), 5.29 (2H, dd, J=9.2, 6.7
Hz), 4.60 (2H, d, J=6.5 Hz), 3.84-3.92 (2H, m), 3.68 (6H, s),
2.69-2.80 (2H, m), 2.51-2.62 (2H, m), 2.07-2.27 (4H, m), 1.10-1.21
(2H, m), 1.02 (6H, d, J=6.8 Hz), 0.95-1.00 (2H, m), 0.93 (6H, d,
J=6.8 Hz).
##STR00647##
Example 48
Dimethyl
(1,5-naphthyridine-2,6-diylbis(1H-benzimidazole-5,2-diyl(1R,3S,5R-
)-2-azabicyclo[3.1.0]hexane-3,2-diyl((1S)-2-oxo-1-(tetrahydro-2H-pyran-4-y-
l)-2,1-ethanediyl)))biscarbamate
[0958] HATU (8.68 mg, 0.023 mmol) was added to a solution of a TFA
salt of
2,6-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol--
6-yl)-1,5-naphthyridine (12 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (5.0 mg, 0.023 mmol) in DMF (0.5 mL) and DIPEA (0.016 mL,
0.089 mmol) and the mixture was stirred at rt for 3 h. The reaction
was diluted with MeOH and purified by prep HPLC (H.sub.2O-MeOH with
0.1% TFA buffer) to yield a TFA salt of dimethyl
(1S,1'S)-2,2'-((1R,1'R,3S,3'S,5R,5'R)-3,3'-(6,6'-(1,5-naphthyridine-2,6-d-
iyl)bis(1H-benzo[d]imidazole-6,2-diyl))bis(2-azabicyclo[3.1.0]hexane-3,2-d-
iyl))bis(2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethane-2,1-diyl)dicarbamate
(11 mg) as yellow solid. LC-MS retention time 1.73 min; m/z 462
[1/2 M+H].sup.+. (Column PHENOMENEX.RTM.Luna3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.61-8.68 (4H, m), 8.50 (2H, dd, J=8.8, 1.51 Hz), 8.45
(2H, d, J=8.8 Hz), 7.93 (2H, d, J=8.8 Hz), 5.28 (2H, dd, J=9.3, 6.8
Hz), 4.64 (2H, d, J=7.5 Hz), 3.84-4.02 (6H, m), 3.69 (6H, s),
3.35-3.46 (4H, m), 2.75 (2H, dd, J=13.7, 9.2 Hz), 2.58 (2H, d,
J=6.8 Hz), 1.99-2.22 (4H, m), 1.36-1.69 (8H, m), 1.07-1.19 (2H, m),
0.96 (2H, m).
##STR00648##
Example 49
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((me-
thoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imi-
dazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-
carbonyl)-2-methylpropyl)carbamate
[0959] HATU (76 mg, 0.20 mmol) was added to a solution of
2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-5-(6-(2-((1R,3S,5R)-2-azabicyc-
lo[3.1.0]hex-3-yl)-4-chloro-1H-imidazol-5-yl)-2-naphthyl)-1H-benzimidazole
(34 mg), (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (35.2
mg, 0.201 mmol) and DIEA (0.12 mL, 0.67 mmol) in DMF (3 mL) and the
resulting mixture was stirred at ambient temperature for 2 hours.
The reaction mixture was diluted with methanol (2 mL) and water (2
mL) and the volatile component was removed in vacuo. The residue
was purified by a reverse phase preparative HPLC (water/methanol,
0.1% TFA) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbon-
yl)-2-methylpropyl)carbamate (41.3 mg) as yellow solid. LC-MS
retention time 1.717 min; m/z 821.58 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire C18 4.6.times.30 mm 5 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% methanol/90% water/0.1% TFA and Solvent B
was 10% water/90% methanol/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(500 MHz, MeOD) .delta. ppm 8.24 (d, J=1.8 Hz, 2H), 8.01-8.13 (m,
4H), 7.84-7.98 (m, 3H), 5.29 (dd, J=9.2, 6.7 Hz, 1H), 5.09 (t,
J=7.2 Hz, 1H), 4.60 (t, J=7.2 Hz, 2H), 3.86-3.91 (m, 1H), 3.71-3.76
(m, 1H), 3.69 (s, 3H), 3.68 (m, 3H), 2.77 (dd, J=13.6, 9.3 Hz, 1H),
2.57 (ddd, J=13.6, 6.7, 6.6 Hz, 1H), 2.44-2.53 (m, 2H), 2.11-2.26
(m, 3H), 1.99-2.10 (m, 1H), 1.10-1.22 (m, 2H), 0.89-1.09 (m, 13H),
0.80-0.84 (m, 1H).
##STR00649##
Example 50
Methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(2-((1R,3S,5R)-2-((2S)-2-((met-
hoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0-
]hex-3-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicycl-
o[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0960] HATU (45.0 mg, 0.118 mmol) was added to a solution of
2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-5-(6-(2-((1R,3S,5R)-2-azabicyc-
lo[3.1.0]hex-3-yl)-4-chloro-1H-imidazol-5-yl)-2-naphthyl)-1H-benzimidazole
(20 mg, 0.039 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (25.7 mg) and DIEA (0.069 mL, 0.39 mmol) in DMF (3 mL) and the
resulting mixture was stirred at ambient temperature for 2 hours.
The reaction mixture was diluted with methanol (2 mL) and water (2
mL) and the volatile component was removed in vacuo. The residue
was purified by a reverse phase preparative HPLC (water/methanol,
0.1% TFA) to yield a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-
-yl)-1H-benzimidazol-5-yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.-
0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (28.1
mg) as pale yellow solid. LC-MS retention time 1.597 min; m/z
905.67 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire C18 4.6.times.30 mm 5
mm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 4
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 2 min, a hold time of 1 min,
and an analysis time of 3 min where Solvent A was 10% methanol/90%
water/0.1% TFA and Solvent B was 10% water/90% methanol/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.24 (s,
2H), 8.02-8.13 (m, 4H), 7.86-7.97 (m, 3H), 5.27 (dd, J=9.2, 7.0 Hz,
1H), 5.08 (t, J=7.2 Hz, 1H), 4.65 (t, J=7.3 Hz, 2H), 3.88-4.02 (m,
5H), 3.75-3.83 (m, 1H), 3.70 (s, 3H), 3.68 (s, 3H), 3.36-3.49 (m,
4H), 2.77 (dd, J=13.7, 9.5 Hz, 1H), 2.57 (ddd, J=13.7, 6.7, 6.4 Hz,
1H), 2.46-2.53 (m, 2H), 2.00-2.20 (m, 4H), 1.55-1.69 (m, 4H),
1.40-1.70 (m, 4H), 1.08-1.20 (m, 2H), 0.94-1.00 (m, 1H), 0.80-0.84
(m, 1H).
##STR00650##
Example 51
Methyl
((1S)-1-(((1S,3R,5S)-3-(4-(6-(3-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)car-
bonyl)-2-methylpropyl)carbamate
[0961] HATU (47.8 mg, 0.126 mmol) was added to a mixture of a TFA
salt of
(1R,3S,5R)-3-(4-(3-(6-(2-((1S,3R,5S)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imid-
azol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(40 mg), (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (22 mg,
0.13 mmol) and DIEA (0.073 mL, 0.42 mmol) in DMF (3 mL) and the
resulting mixture was stirred at ambient temperature for 2 hours.
The reaction mixture was diluted with methanol (2 mL) and water (2
mL) and the volatile component was removed in vacuo. The residue
was partially purified by a reverse phase HPLC (water/methanol,
0.1% TFA) and then repurified by reverse phase HPLC
(water/acetonitrile, 0.1% TFA) to yield a TFA salt of methyl
((1S)-1-(((1S,3R,5S)-3-(4-(6-(3-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)p-
henyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate (45 mg) as a white solid. LC-MS retention
time 1.458 min; m/z 813.30 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire C18 4.6.times.30 mm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions
employed a flow rate of 4 mL/min, a gradient of 100% Solvent A/0%
Solvent B to 0% Solvent A/100% Solvent B, a gradient time of 2 min,
a hold time of 1 min, and an analysis time of 3 min where Solvent A
was 10% methanol/90% water/10 mM ammonium acetate and Solvent B was
10% water/90% methanol/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.30 (d, J=4.6 Hz,
2H), 8.18 (s, 1H), 8.14 (d, J=8.9 Hz, 1H), 8.10 (d, J=8.9 Hz, 1H),
7.98-8.02 (m, 2H), 7.98 (s, 1H), 7.93 (d, J=8.2 Hz, 1H), 7.88 (dd,
J=8.7, 1.7 Hz, 1H), 7.77 (d, J=8.2 Hz, 1H), 7.69 (t, J=7.8 Hz, 1H),
5.13-5.20 (m, 2H), 4.57 (t, J=6.1 Hz, 2H), 3.80-3.88 (m, 2H), 3.69
(d, J=6.1 Hz, 6H), 2.67-2.77 (m, 2H), 2.51 (dq, J=13.7, 6.8 Hz,
2H), 2.15-2.25 (m, 2H), 2.10 (dq, J=13.8, 6.8 Hz, 2H), 1.08-1.18
(m, 2H), 1.03 (dd, J=6.7, 4.6 Hz, 6H), 0.95 (dd, J=6.7, 3.4 Hz,
6H), 0.92 (dd, J=5.3, 2.0 Hz, 2H).
##STR00651##
Example 52
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(2-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-
-5-yl)-6-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbo-
nyl)-2-methylpropyl)carbamate
[0962] HATU (13.16 mg, 0.035 mmol) was added to a solution of a TFA
salt of
2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6--
yl)-6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)quino-
xaline (14 mg, 0.015 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (6.1 mg) in DMF
(0.5 mL) and DIPEA (0.024 mL, 0.135 mmol) and the mixture was
stirred at rt for 16 h. The reaction was diluted with MeOH and
purified by prep HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield
a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(2-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-6-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate (11.8 mg,) as a yellow solid. LC-MS
retention time 1.77 min; m/z 789 [M+H].sup.+. (Column
PHENOMENEX.RTM. Luna3.0.times.50 mm S10. Solvent A=90% water:10%
methanol: 0.1% TFA. Solvent B=10% water:90% methanol: 0.1% TFA.
Flow Rate=4 mL/min. Start % B=0. Final % B=100. Gradient Time=3
min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.59
(s, 1H), 8.67 (s, 1H), 8.51 (d, J=8.8 Hz, 1H), 8.46 (d, J=1.8 Hz,
1H), 8.27 (d, J=8.8 Hz, 1H), 8.19 (dd, J=8.8, 2.0 Hz, 1H), 8.11 (s,
1H), 7.92 (d, J=8.5 Hz, 1H), 5.29 (dd, J=9.2, 6.7 Hz, 1H), 5.17
(dd, J=9.2, 7.2 Hz, 1H), 4.58 (dd, J=13.2, 6.7 Hz, 2H), 3.78-3.92
(m, 2H), 3.68 (s, 6H), 2.67-2.79 (m, 2H), 2.45-2.62 (m, 2H),
2.03-2.27 (m, 4H), 1.07-1.21 (m, 2H), 1.03 (d, J=6.8 Hz, 6H),
0.93-1.00 (m, 2H), 0.93 (dd, J=6.3, 5.8 Hz, 6H).
##STR00652##
Example 53
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(2-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl-
)-1H-benzimidazol-5-yl)-6-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0963] HATU (15.0 mg, 0.040 mmol) was added to a solution of a TFA
salt of
2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-yl)-
-6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)quinoxal-
ine (16 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (8.6 mg, 0.040 mmol) in DMF (0.5 mL) and DIPEA (0.027 mL, 0.16
mmol) and the mixture was stirred at rt for 16 h. The reaction was
diluted with MeOH, and purified by prep HPLC (H.sub.2O-MeOH with 10
mM NH.sub.4OAc buffer) to yield methyl
((1S)-2-((1R,3S,5R)-3-(4-(2-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-b-
enzimidazol-5-yl)-6-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (14.5 mg)
as yellow solid. LC-MS retention time 1.58 min; m/z 873
[M+H].sup.+. (Column PHENOMENEX.RTM. Luna3.0.times.50 mm S10.
Solvent A=90% water:10% methanol: 0.1% TFA. Solvent B=10% water:90%
methanol: 0.1% TFA. Flow Rate=4 mL/min. Start % B=0. Final % B=100.
Gradient Time=3 min. Wavelength=220). .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 9.34 (s, 1H), 8.36 (s, 1H), 8.28 (d, J=1.0 Hz, 1H),
8.08-8.19 (m, 2H), 8.03 (d, J=8.8 Hz, 1H), 7.66 (d, J=8.5 Hz, 1H),
7.54 (s, 1H), 5.27 (t, J=6.9 Hz, 1H), 5.18 (dd, J=8.9, 4.9 Hz, 1H),
4.69 (d, J=5.5 Hz, 2H), 3.87-4.03 (m, 4H), 3.73-3.85 (m, 2H), 3.68
(s, 6H), 3.32-3.47 (m, 4H), 2.50-2.61 (m, 3H), 2.40-2.49 (m, 1H),
1.99-2.16 (m, 4H), 1.51-1.72 (m, 6H), 1.38-1.51 (m, 2H), 1.09-1.22
(m, 2H), 0.76-0.92 (m, 2H).
##STR00653##
Example 54
Methyl
((1S)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl-
)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(-
tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0964] HATU (157 mg, 0.413 mmol) was added to a solution of methyl
(S)-1-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1-
H-benzo[d]imidazol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (130 mg),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (90 mg, 0.41 mmol) and DIEA (0.36 mL, 2.1 mmol) in DMF (4 mL)
and the reaction was stirred 2 h at room temperature. The reaction
mixture was diluted with MeOH (2 mL) and water (2 mL). The
volatiles were removed under vacuum. The material was purified on
reverse phase HPLC (water/methanol, 0.1% TFA) to afford a TFA salt
of methyl
((1S)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-na-
phthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrah-
ydro-2H-pyran-4-yl)ethyl)carbamate (194 mg) as yellow solid. LC-MS
retention time 1.307 min; m/z 829.27 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire C18 4.6.times.30 mm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions
employed a flow rate of 4 mL/min, a gradient of 100% Solvent A/0%
Solvent B to 0% Solvent A/100% Solvent B, a gradient time of 2 min,
a hold time of 1 min, and an analysis time of 3 min where Solvent A
was 10% methanol/90% water/0.1% TFA and Solvent B was 10% water/90%
methanol/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (TFA salt, 500
MHz, MeOD) .delta. ppm 8.31 (s, 1H), 8.29 (s, 1H), 8.16 (d, J=8.5
Hz, 1H), 8.11 (d, J=0.9 Hz, 1H), 8.11 (d, J=8.9 Hz, 1H), 8.02 (dd,
J=8.5, 1.5 Hz, 1H), 8.00 (s, 1H), 7.98 (dd, J=8.5, 1.5 Hz, 1H),
7.89 (d, J=8.5 Hz, 1H), 7.88 (dd, J=8.5, 1.8 Hz, 1H), 5.27 (dd,
J=9.2, 6.7 Hz, 1H), 5.18 (dd, J=9.2, 7.0 Hz, 1H), 4.62-4.67 (m,
1H), 4.58 (d, J=6.4 Hz, 1H), 3.87-4.01 (m, 3H), 3.82-3.87 (m, 1H),
3.70 (m, 6H), 3.36-3.44 (m, 2H), 2.74 (td, J=13.3, 9.5 Hz, 2H),
2.48-2.62 (m, 2H), 2.04-2.24 (m, 4H), 1.40-1.60 (m, 4H), 1.09-1.19
(m, 2H), 1.04 (d, J=6.7 Hz, 3H), 0.89-0.99 (m, 5H).
##STR00654##
Example 55
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-y-
l)-1H-imidazol-4-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]-
hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[0965] HATU (100 mg, 0.263 mmol) was added to a solution of methyl
(S)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1-
H-benzo[d]imidazol-6-yl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexan-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethylcarbamate
(118 mg), (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (46.2
mg, 0.263 mmol) and DIEA (0.31 mL, 1.8 mmol) in DMF (4 mL) and the
reaction was stirred 2 h at room temperature. The reaction mixture
was diluted with MeOH (2 mL) and water (2 mL). The volatiles were
removed under vacuum using a rotavap and the residue was purified
on reverse phase HPLC (water/methanol, 0.1% TFA) to afford the a
TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2--
yl)carbonyl)-2-methylpropyl)carbamate (55.3 mg). LC-MS retention
time 1.320 min; m/z 829.42 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire C18 4.6.times.30 mm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions
employed a flow rate of 4 mL/min, a gradient of 100% Solvent A/0%
Solvent B to 0% Solvent A/100% Solvent B, a gradient time of 2 min,
a hold time of 1 min, and an analysis time of 3 min where Solvent A
was 10% methanol/90% water/0.1% TFA and Solvent B was 10% water/90%
methanol/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (TFA salt, 500
MHz, MeOD) .delta. ppm 8.31 (s, 1H) 8.29 (s, 1H), 8.08-8.18 (m,
3H), 8.04 (dd, J=8.9, 1.5 Hz, 1H), 7.95-8.00 (m, 2H), 7.90 (d,
J=8.6 Hz, 1H), 7.87 (dd, J=8.6, 1.8 Hz, 1H), 5.29 (dd, J=9.3, 6.9
Hz, 1H), 5.17 (dd, J=9.2, 7.0 Hz, 1H), 4.62 (dd, J=9.2, 7.3 Hz,
2H), 3.96 (td, J=10.8, 2.9 Hz, 2H), 3.83-3.92 (m, 2H), 3.64-3.77
(m, 6H), 3.35-3.46 (m, 2H), 2.74 (ddd, J=19.3, 13.5, 9.3 Hz, 2H),
2.48-2.62 (m, 2H), 2.06-2.26 (m, 4H), 1.45-1.63 (m, 4H), 1.06-1.22
(m, 2H), 1.03 (d, J=6.7 Hz, 3H), 0.96-1.01 (m, 1H), 0.93 (d, J=6.7
Hz, 3H), 0.88-091 (m, 1H).
##STR00655##
Example 56
Methyl
((1S)-2-((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((met-
hoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imid-
azol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)--
2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0966] NCS (14.98 mg, 0.112 mmol) was added to a solution of methyl
((1S)-2-((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-na-
phthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrah-
ydro-2H-pyran-4-yl)ethyl)carbamate (93 mg) in DMF and the reaction
mixture was heated at 50.degree. C. (oil bath temp) for 2 h. The
crude reaction mixture was purified on reverse phase HPLC
(water/methanol, 0.1% TFA) to afford a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-5-
-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo--
1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (102.8 mg). LC-MS
retention time 1.648 min; m/z 863.32 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a Waters
Sunfire C18 4.6.times.30 mm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions
employed a flow rate of 4 mL/min, a gradient of 100% Solvent A/0%
Solvent B to 0% Solvent A/100% Solvent B, a gradient time of 2 min,
a hold time of 1 min, and an analysis time of 3 min where Solvent A
was 10% methanol/90% water/0.1% TFA and Solvent B was 10% water/90%
methanol/0.1% TFA. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (TFA salt, 500
MHz, MeOD) .delta. ppm 8.22 (br s, 2H), 8.10 (s, 1H), 8.00-8.06 (m,
3H), 7.85-7.94 (m, 3H), 5.28 (dd, J=9.2, 7.0 Hz, 1H), 5.09 (t,
J=7.2 Hz, 1H), 4.65 (d, J=7.3 Hz, 1H), 4.59 (d, J=7.0 Hz, 1H),
3.89-3.99 (m, 3H), 3.66-3.77 (m, 7H), 3.35-3.44 (m, 2H), 2.77 (dd,
J=13.6, 9.3 Hz, 1H), 2.57 (ddd, J=13.8, 6.8, 6.6 Hz, 1H), 2.47-2.52
(m, 2H), 2.02-2.22 (m, 4H), 1.44-1.63 (m, 4H), 1.13 (dt, J=8.6, 5.8
Hz, 2H), 1.05 (d, J=7.0 Hz, 3H), 0.92-1.02 (m, 4H), 0.83 (d, J=1.8
Hz, 1H).
##STR00656##
Example 57
Methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((me-
thoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.-
0]hex-3-yl)-1H-imidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[0967] NCS (10.31 mg, 0.077 mmol) was added to a solution of methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1.0]hex-2--
yl)carbonyl)-2-methylpropyl)carbamate (64 mg) in DMF (2 mL) and the
reaction mixture was heated at 50.degree. C. for 2 h. The crude
reaction mixture was purified on reverse phase HPLC
(water/methanol, 0.1% TFA) and repurified on reverse phase HPLC
(water/ACN, 0.1% TFA) to afford the TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(5-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex--
3-yl)-1H-imidazol-5-yl)-2-naphthyl)-1H-benzimidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate (44.7 mg) as tan
solid. LC-MS retention time 1.63 min; m/z 863.39 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Waters Sunfire C18 4.6.times.30 mm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% methanol/90% water/0.1% TFA and Solvent B
was 10% water/90% methanol/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.24 (br s, 2H), 8.02-8.12
(m, 4H), 7.86-7.95 (m, 3H), 5.28 (dd, J=9.2, 6.7 Hz, 1H), 5.08 (t
apparent, J=7.0 Hz, 1H), 4.65 (d, J=7.6 Hz, 1H), 4.60 (d, J=6.7 Hz,
1H), 3.93-4.01 (m, 2H), 3.86-3.91 (m, 1H), 3.75-3.81 (m, 1H),
3.65-3.74 (m, 6H), 3.36-3.46 (m, 2H), 2.77 (dd, J=13.7, 9.2 Hz,
1H), 2.57 (ddd, J=13.7, 6.7, 6.4 Hz, 1H), 2.49 (dd, J=7.8, 3.2 Hz,
2H), 2.12-2.25 (m, 2H), 2.00-2.12 (m, 2H), 1.54-1.67 (m, 3H),
1.41-1.51 (m, 1H), 1.09-1.19 (m, 2H), 0.97-1.06 (m, 4H), 0.94 (d,
J=6.7 Hz, 3H), 0.82 (br s, 1H).
##STR00657##
Example 58
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl-
)-1H-benzimidazol-5-yl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0968] HATU (5.64 mg, 0.015 mmol) was added to a solution of a TFA
salt of
6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-yl)-
-2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)quinoxal-
ine (6.0 mg) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (3.2 mg) in DMF (0.5 mL) and DIPEA (10.13 .mu.L, 0.058 mmol)
and the mixture was stirred at rt for 16 h. The solvent was removed
and the residue was purified by prep HPLC (H.sub.2O-MeOH with 0.1%
TFA buffer) to yield a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-b-
enzimidazol-5-yl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (5.5 mg)
as a yellow solid. LC-MS retention time 1.60 min; m/z 873
[M+H].sup.+. (Column PHENOMENEX.RTM. Luna3.0.times.50 mm S10.
Solvent A=95% water/5% methanol/10 mM ammonium acetate. Solvent
B=5% water/95% methanol/10 mM ammonium acetate. Flow Rate=4 mL/min.
Start % B=0. Final % B=100. Gradient Time=2 min. Wavelength=220).
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.42 (s, 1H), 8.41-8.50 (m,
2H), 8.30 (d, J=1.3 Hz, 2H), 8.19 (d, J=1.0 Hz, 1H), 8.07 (dd,
J=8.5, 1.5 Hz, 1H), 7.92 (d, J=8.5 Hz, 1H), 5.13-5.33 (m, 3H),
4.57-4.70 (m, 1H), 3.81-4.04 (m, 6H), 3.62-3.77 (m, 6H), 3.34-3.46
(m, 4H), 2.65-2.82 (m, 2H), 2.45-2.61 (m, 2H), 2.01-2.22 (m, 4H),
1.40-1.65 (m, 8H), 1.03-1.17 (m, 2H), 0.83-0.98 (m, 2H).
##STR00658##
Example 59
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-
-5-yl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbo-
nyl)-2-methylpropyl)carbamate
[0969] HATU (5.64 mg, 0.015 mmol) was added to a solution of a TFA
salt of
6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-benzo[d]imidazol-6-yl)-
-2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)quinoxal-
ine (6.0 mg) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid
(2.60 mg, 0.015 mmol) in DMF (0.5 mL) and DIPEA (10.13 .mu.L, 0.058
mmol) and the mixture was stirred at rt for 16 h. The solvent was
removed and the residue was purified by prep HPLC (H.sub.2O-MeOH
with 0.1% TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-benzimidazol-5-yl)-
-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate (3.6 mg) as yellow solid. LC-MS retention
time 1.75 min; m/z 789 [M+H].sup.+. (Column PHENOMENEX.RTM.
Luna3.0.times.50 mm S10. Solvent A=90% water:10% methanol: 0.1%
TFA. Solvent B=10% water:90% methanol: 0.1% TFA. Flow Rate=4
mL/min. Start % B=0. Final % B=100. Gradient Time=3 min.
Wavelength=220). .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.42 (s,
1H), 8.41-8.48 (m, 2H), 8.30 (d, J=1.3 Hz, 2H), 8.18 (d, J=1.0 Hz,
1H), 8.06 (dd, J=8.7, 1.6 Hz, 1H), 7.92 (d, J=8.5 Hz, 1H),
5.24-5.33 (m, 1H), 5.16-5.24 (m, 1H), 4.52-4.63 (m, 2H), 3.78-3.91
(m, 2H), 3.68 (d, J=1.0 Hz, 6H), 2.65-2.79 (m, 2H), 2.46-2.61 (m,
2H), 2.05-2.26 (m, 4H), 1.07-1.20 (m, 2H), 0.99-1.06 (m, 6H),
0.94-1.01 (m, 2H), 0.92 (dd, J=6.8, 3.3 Hz, 6H).
##STR00659##
Example 60
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2--
((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex--
3-yl)-1H-imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[-
3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
##STR00660##
[0970] Example 62
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-(-
(methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)carbonyl)-2-methylpropyl)carbamate
##STR00661##
[0971] Example 63
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-(-
(methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)carbonyl)-2-methylpropyl)carbamate
[0972] NCS (13.0 mg, 0.097 mmol) was added to a solution of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)--
2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate (79 mg) in DMF (2 mL) at room temperature
and the mixture was heated at 50.degree. C. for 16 h. The crude
product was purified by prep HPLC (TFA) using a Waters Sunfire C18
column 30.times.150 mm 5u eluted with a gradient of 30 to 90%
MeOH-Water+0.1% TFA to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)--
2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)--
1H-imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]-
hex-2-yl)carbonyl)-2-methylpropyl)carbamate (20.2 mg) as yellow
solid and a mixture of the two monochloride products which were
separated by SFC on a CHIRALPAK.RTM. IB column and then repurified
individually by prep HPLC (TFA) using a Waters Sunfire C18 column
30.times.150 mm 5u eluted with a gradient of 25 to 100%
MeOH-Water+0.1% TFA to yield a TFA of methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-
carbonyl)-2-methylpropyl)carbamate (11.1 mg) as a yellow solid and
a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-
carbonyl)-2-methylpropyl)carbamate (11.4 mg) as a yellow solid.
[0973] For Example 60: LC-MS retention time 2.133 min; m/z 883.35
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.22 (s,
2H), 8.07 (d, J=8.5 Hz, 1H), 8.03 (d, J=8.9 Hz, 1H), 7.83-7.95 (m,
6H), 5.08 (td, J=9.1, 6.0 Hz, 2H), 4.59 (dd, J=6.7, 1.8 Hz, 2H),
3.71-3.79 (m, 2H), 3.68 (s, 6H), 2.44-2.57 (m, 4H), 2.13-2.25 (m,
2H), 2.00-2.10 (m, 2H), 1.09-1.16 (m, 2H), 1.05 (dd, J=6.7, 1.5 Hz,
6H), 0.91-1.02 (m, 6H), 0.79-0.87 (m, 2H).
[0974] For Example 62: LC-MS retention time 1.745 min; m/z 847.33
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.23 (s,
2H), 8.02-8.09 (m, 2H), 7.98 (d, J=8.2 Hz, 2H), 7.89-7.94 (m, 3H),
7.86 (d, J=8.6 Hz, 2H), 5.15 (dd, J=9.2, 7.0 Hz, 1H), 5.09 (t,
J=7.0 Hz, 1H), 4.58 (t, J=6.7 Hz, 2H), 3.84 (t, J=4.7 Hz, 1H),
3.71-3.77 (m, 1H), 3.69 (d, J=3.1 Hz, 6H), 2.72 (dd, J=13.7, 9.5
Hz, 1H), 2.45-2.54 (m, 3H), 2.14-2.24 (m, 2H), 2.00-2.14 (m, 2H),
1.09-1.18 (m, 2H), 1.00-1.08 (m, 6H), 0.96 (t, J=7.6 Hz, 6H), 0.92
(br s, 1H), 0.83 (br s, 1H).
[0975] For Example 63: LC-MS retention time 1.753 min; m/z 847.32
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Waters Sunfire 5u C18 4.6.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 4 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% TFA and Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.26 (d,
J=7.0 Hz, 2H), 8.13 (d, J=8.9 Hz, 1H), 8.04-8.10 (m, 1H), 7.95-8.01
(m, 2H), 7.89-7.95 (m, 2H), 7.82-7.89 (m, 3H), 5.17 (dd, J=9.2, 7.0
Hz, 1H), 5.07 (t, J=7.2 Hz, 1H), 4.58 (d, J=6.4 Hz, 2H), 3.81-3.88
(m, 1H), 3.71-3.76 (m, 1H), 3.69 (d, J=4.6 Hz, 6H), 2.73 (dd,
J=13.4, 9.5 Hz, 1H), 2.44-2.56 (m, 3H), 2.14-2.25 (m, 2H),
2.08-2.14 (m, 1H), 1.09-1.18 (m, 2H,) 1.00-1.08 (m, 6H), 0.96 (t,
J=6.4 Hz, 6H), 0.92 (br s, 1H) 0.78-0.85 (m, 1H).
##STR00662##
Example 61
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-
-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-o-
xo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0976] The reaction of Intermediate 105 and the appropriate boronic
acid (prepared in a similar manner as Intermediate 66 using the
appropriate starting materials) via a coupling reaction similar to
the preparation of Intermediate 106 yielded a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ph-
enyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(-
tetrahydro-2H-pyran-4-yl)ethyl)carbamate (91 mg) as a pale yellow
solid. LC-MS retention time 1.362 min; m/z 855.31 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a Waters Sunfire 5u C18 4.6.times.30 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time
of 3 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1% TFA and
Solvent B was 10% H.sub.2O/90% MeOH/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.29 (s, 1H), 8.28
(s, 1H), 8.14 (d, J=8.9 Hz, 1H), 8.09 (d, J=8.9 Hz, 1H), 8.01-7.94
(m, 4H), 7.93-7.91 (m, 1H), 7.90-7.83 (m, 3H), 5.19-5.12 (m, 2H),
4.62 (d, J=7.6 Hz, 1H), 4.58 (d, J=6.4 Hz, 1H), 3.99-3.91 (m, 2H),
3.89-3.80 (m, 2H), 3.70 (s, 3H), 3.69 (s, 3H), 3.46-3.35 (m, 2H),
2.76-2.67 (m, 2H), 2.57-2.46 (m, 2H), 2.25-2.16 (m, 1H), 2.15-2.05
(m, 3H), 1.65-1.40 (m, 4H), 1.16-1.07 (m, 2H), 1.04 (d, J=6.7 Hz,
3H), 0.95 (d, J=6.7 Hz, 3H), 0.93-0.87 (m, 2H).
##STR00663##
Example 64
Methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-(-
(2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-
-yl)-1H-imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0977] N-Chlorosuccinimide (10.4 mg, 0.078 mmol) was added to a
stirred solution of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ph-
enyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(-
tetrahydro-2H-pyran-4-yl)ethyl)carbamate (Example 61) (60 mg, 0.071
mmol) in DMF (1.5 mL) and the reaction vessel was sealed and heated
at 50.degree. C. for 16 h. The reaction was cooled, partially
concentrated and then diluted with MeOH. The solution was filtered
and purified by preparative HPLC (MeOH/H.sub.2O, w 0.1% TFA) to
yield a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2--
((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex--
3-yl)-1H-imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[-
3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
(21.4 mg) as a yellow solid. LC-MS retention time 2.798 min; m/z
462.34 (1/2MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.18 (s, 2H), 8.03 (d,
J=8.8 Hz, 1H), 7.99 (d, J=8.8 Hz, 1H), 7.80-7.92 (m, 6H), 5.06
(ddd, J=9.0, 5.8, 3.5 Hz, 2H), 4.63 (d, J=7.5 Hz, 1H), 4.57 (d,
J=6.5 Hz, 1H), 3.92-3.98 (m, 2H), 3.71-3.81 (m, 2H), 3.67 (s, 6H),
3.35-3.44 (m, 2H), 2.42-2.57 (m, 4H), 1.99-2.24 (m, 4H), 1.40-1.66
(m, 4H), 1.06-1.14 (m, 2H), 1.03 (d, J=6.8 Hz, 3H), 0.95 (d, J=6.8
Hz, 3H), 0.82 (d, J=2.8 Hz, 2H).
##STR00664##
Example 65
Methyl
((1S)-1-(4,4-difluorocyclohexyl)-2-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3-
S,5R)-2-((2S)-2-(4,4-difluorocyclohexyl)-2-((methoxycarbonyl)amino)acetyl)-
-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imid-
azol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxoethyl)carbamate
[0978] HATU (67.3 mg, 0.177 mmol) was added to a stirred solution
of an HCl salt of
(1R,3S,5R)-3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-im-
idazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
exane (Intermediate 69) (38.0 mg, 0.059 mmol) and
(S)-2-(4,4-difluorocyclohexyl)-2-(methoxycarbonylamino)acetic acid
(44.5 mg, 0.177 mmol) in DMF (0.7 mL) and DIPEA (0.103 mL, 0.590
mmol). The reaction was diluted with MeOH, filtered, purified by
preparative HPLC (water/MeOH w/0.1% TFA) and repurified by
preparative HPLC (water/MeOH w/10 mM ammonium acetate) to yield
methyl
((1S)-1-(4,4-difluorocyclohexyl)-2-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)--
2-((2S)-2-(4,4-difluorocyclohexyl)-2-((methoxycarbonyl)amino)acetyl)-2-aza-
bicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-
-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxoethyl)carbamate (22.1 mg) as
a light yellow solid. LC-MS retention time 2.713 min; m/z 483.13
(1/2MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.16 (s, 1H), 8.11 (s, 1H),
7.94 (d, J=8.5 Hz, 2H), 7.77-7.85 (m, 5H), 7.47 (s, 1H), 7.39 (s,
1H), 7.08 (d, J=8.5 Hz, 1H), 5.15 (ddd, J=8.2, 6.3, 6.1 Hz, 2H),
4.68 (br. s., 2H), 3.69-3.76 (m, 2H), 3.68 (s, 6H), 2.41-2.57 (m,
4H), 1.65-2.12 (m, 16H), 1.33-1.59 (m, 4H), 1.06-1.17 (m, 2H), 0.78
(br. s., 2H).
##STR00665##
Example 66
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)butanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-n-
aphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)pro-
pyl)carbamate
[0979] HATU (89 mg, 0.24 mmol) was added to a stirred solution of a
TFA salt of
(1R,3S,5R)-3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-y-
l)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo-
[3.1.0]hexane (Intermediate 69) (80 mg, 0.084 mmol) and
(S)-2-(methoxycarbonylamino)butanoic acid (37.8 mg, 0.235 mmol) in
DMF (0.8 mL) and DIPEA (0.15 mL, 0.84 mmol). The reaction was
diluted with MeOH, filtered and purified by preparative HPLC
(water/MeOH w/0.1% TFA) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)butanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthy-
l)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)propyl)ca-
rbamate (70.1 mg) as a light yellow solid. LC-MS retention time
1.688 min; m/z 785.55 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.28 (s, 1H), 8.26 (s, 1H),
8.12 (d, J=8.8 Hz, 1H), 8.07 (d, J=8.5 Hz, 1H), 7.93-8.00 (m, 4H),
7.82-7.90 (m, 4H), 5.12-5.21 (m, 2H), 4.58-4.65 (m, 2H), 3.74-3.81
(m, 2H), 3.67 (s, 6H), 2.70 (ddd, J=13.7, 9.0, 4.9 Hz, 2H), 2.49
(dq, J=14.2, 7.1 Hz, 2H), 2.09 (br. s., 2H), 1.86-1.99 (m, 2H),
1.64-1.77 (m, 2H), 1.09-1.17 (m, 2H), 1.02 (t, J=7.4 Hz, 6H), 0.92
(br. s., 2H).
##STR00666##
Example 67
Methyl
((1S)-1-(((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methy-
lpropyl)carbamate
[0980] HATU (101 mg, 0.266 mmol) was added to a stirred solution of
a TFA salt of
2-((2S,5S)-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl--
2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 117) (102 mg, 0.106 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (46.6 mg, 0.266
mmol) in DMF (1.0 mL) and DIPEA (0.189 mL, 1.1 mmol) and the
reaction was stirred at rt for 2 h. The reaction mixture was
diluted with MeOH and purified by preparative HPLC (MeOH/H.sub.2O
w/0.1% TFA) and then repurified by preparative HPLC
(acetonitrile/water with 0.1% TFA) to yield a TFA salt of methyl
((1S)-1-(((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate (63.8 mg) as a light yellow solid. LC-MS retention time
1.900 min; m/z 817.59 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
10u C18 3.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR presents as a .about.4:1 mixture of regioisomers.
.sup.1H NMR (for major regioisomer) (400 MHz, MeOD) .delta. ppm
8.31 (s, 1H), 8.27 (s, 1H), 8.13 (d, J=8.5 Hz, 1H), 8.09 (d, J=8.5
Hz, 1H), 7.94-8.02 (m, 5H), 7.84-7.92 (m, 3H), 5.15-5.24 (m, 2H),
4.13 (dd, J=8.7, 1.4 Hz, 2H), 3.74 (d, J=2.8 Hz, 2H), 3.67 (s, 6H),
2.50-2.60 (m, 2H), 2.26-2.45 (m, 4H), 1.95-2.10 (m, 4H), 1.57 (dd,
J=6.5, 3.0 Hz, 6H), 1.32 (t, J=6.4 Hz, 2H), 0.99 (d, J=6.8 Hz, 6H),
0.89 (d, J=6.8 Hz, 6H).
##STR00667##
Example 68
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-(-
(methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3-
.1.0]hex-3-yl)-1H-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-az-
abicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate and
##STR00668##
[0981] Example 69
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-(-
(methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3-
.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-az-
abicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[0982] A mixture of two chloroimidazole regioisomers as TFA salts
(Examples 68 and 69) (21.4 mg) was isolated from the same reaction
that prepared a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-
-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (Example
64). The two regioisomeric compounds were separated in multiple
injections using SCF chromatography (2-ethylpyridine SCF column
(4.6.times.250 mm, 5 .mu.m) and 80% CO.sub.2-20% EtOH/0.1%
DEA).
[0983] The absolute regiochemistry of the separated materials was
determined by HMBC correlation. The reaction yielded methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]h-
ex-3-yl)-1H-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate (7.4 mg) and
methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]h-
ex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate (4.5 mg), each
as an off-white solid. For Example 68: LC-MS retention time 2.683
min; m/z 889.17 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode. For
Example 69: LC-MS retention time 2.695 min; m/z 889.22 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 4 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min, and an
analysis time of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10
mM ammonium acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00669##
Example 70
Methyl
((1S)-2-((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-pyrrolidinyl)-1H-im-
idazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-
-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0984] HATU (73.3 mg, 0.193 mmol) was added to a stirred solution
of a TFA salt of
2-((2S,5S)-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl--
2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 117) (73.9 mg, 0.077 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (41.9 mg, 0.193 mmol) in DMF (1.0 mL) and DIPEA (0.14 mL, 0.77
mmol) and the reaction was stirred at rt for 30 min. The reaction
mixture was diluted with MeOH, filtered and purified by preparative
HPLC (MeOH/H.sub.2O w/0.1% TFA) to yield a TFA salt of methyl
((1S)-2-((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-
-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-2-oxo-
-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (51.2 mg) as a yellow
solid. LC-MS retention time 1.897 min; m/z 901.39 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 4 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time
of 3 min where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode.
##STR00670##
Example 71
Methyl
((1S)-1-(4,4-difluorocyclohexyl)-2-((2S,5S)-2-(4-(4-(6-(2-((2S,5S)--
1-((2S)-2-(4,4-difluorocyclohexyl)-2-((methoxycarbonyl)amino)acetyl)-5-met-
hyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-
-5-methyl-1-pyrrolidinyl)-2-oxoethyl)carbamate
[0985] HATU (70.8 mg, 0.186 mmol) was added to a stirred solution
of a TFA salt of
2-((2S,5S)-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl--
2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 117) (71.4 mg, 0.074 mmol) and
(S)-2-(4,4-difluorocyclohexyl)-2-(methoxycarbonylamino)acetic acid
(46.8 mg, 0.186 mmol) in DMF (1.0 mL) and DIPEA (0.13 mL, 0.75
mmol) and the reaction was stirred at rt for 1.5 h. The reaction
was diluted with MeOH, filtered and purified in by preparative HPLC
(MeOH/H.sub.2O w/0.1% TFA) to yield the desired product a TFA salt
of methyl
((1S)-1-(4,4-difluorocyclohexyl)-2-((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S-
)-2-(4,4-difluorocyclohexyl)-2-((methoxycarbonyl)amino)acetyl)-5-methyl-2--
pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-met-
hyl-1-pyrrolidinyl)-2-oxoethyl)carbamate (43.3 mg) as a light
yellow solid. LC-MS retention time 2.828 min; m/z 485.22 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 4 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min, and an
analysis time of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10
mM ammonium acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00671##
Example 72
Methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(4-(6-(4-chloro-2-((1R,3S,5R)-2-(-
(2S)-2-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azab-
icyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2--
yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)c-
arbamate
[0986] N-Chlorosuccinimide (7.1 mg, 0.053 mmol) was added to a
stirred solution of methyl
((1S)-2-((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (Example
34) (22.7 mg, 0.025 mmol) in DMF (0.5 mL) and the reaction vessel
was sealed and the reaction was heated at 50.degree. C. for 12 h.
The reaction was cooled to rt, additional N-chlorosuccinimide
(.about.3 mg) was added and the reaction vessel was sealed and
stirred at 50.degree. C. for 2 h. The reaction was cooled to rt,
diluted with MeOH, filtered and purified by preparative HPLC
(Methanol/Water w/0.1% TFA) to yield methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(4-(6-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-
-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2--
azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbama-
te (12 mg) as a yellow solid. LC-MS retention time 2.708 min; m/z
483.25 (1/2MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 10u C18
3.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 4 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 3 min, a hold time
of 1 min, and an analysis time of 4 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.18 (s, 2H), 8.04 (d,
J=8.8 Hz, 1H), 7.99 (d, J=8.8 Hz, 1H), 7.80-7.92 (m, 6H), 5.02-5.09
(m, 2H), 4.63 (d, J=6.0
[0987] Hz, 2H), 3.91-3.99 (m, 4H), 3.73-3.80 (m, 2H), 3.67 (s, 6H),
3.34-3.45 (m, 4H), 2.43-2.54 (m, 4H), 1.98-2.13 (m, 4H), 1.38-1.67
(m, 8H), 1.06-1.15 (m, 2H), 0.81 (br. s., 2H).
##STR00672##
Example 73
Methyl
((1S,2R)-2-methoxy-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-(N-(m-
ethoxycarbonyl)-O-methyl-L-threonyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imida-
zol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl-
)carbonyl)propyl)carbamate
[0988] HATU (65.7 mg, 0.173 mmol) was added to a stirred solution
of a TFA salt of
2-((2S,5S)-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl--
2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 133) (66 mg, 0.069 mmol) and
(2S,3R)-3-methoxy-2-(methoxycarbonylamino)butanoic acid (33.0 mg,
0.173 mmol) in DMF (0.8 mL) and DIPEA (0.121 mL, 0.691 mmol) and
the reaction was stirred at rt overnight. The reaction was diluted
with MeOH, filtered and purified preparative HPLC (Methanol/water
w/10 nM ammonium acetate) to yield a TFA salt of methyl
((1S,2R)-2-methoxy-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-(N-(methoxy-
carbonyl)-O-methyl-L-threonyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4--
yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbo-
nyl)propyl)carbamate (51.9 mg, 0.048 mmol, 98% yield) as an
off-white solid. LC-MS retention time 2.383 min; m/z 845.42 (MH+).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 10u C18 3.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 4 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 3 min, a hold time of 1 min, and an
analysis time of 4 min where Solvent A was 5% MeOH/95% H.sub.2O/10
mM ammonium acetate and Solvent B was 5% H.sub.2O/95% MeOH/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (MeOD, 400 MHz)
.delta. 8.29 (br s, 1H), 8.26 (s, 1H), 8.12 (d, J=8.8 Hz, 1H), 8.07
(d, J=8.8 Hz, 1H), 8.00-7.93 (m, 4H), 7.91-7.82 (m, 4H), 5.18 (td,
J=9.0, 7.0 Hz, 2H), 4.78 (dd, J=3.9, 1.7 Hz, 2H), 3.92-3.79 (m,
4H), 3.68 (s, 6H), 3.35 (s, 3H), 3.35 (m, 3H), 2.76-2.65 (m, 2H),
2.59-2.47 (m, 2H), 2.15-2.05 (m, 2H), 1.19 (d, J=6.3 Hz, 6H),
1.16-1.07 (m, 2H), 0.94-0.84 (m, 2H).
##STR00673##
Example 74
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4--
yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carb-
onyl)-2-methylpropyl)carbamate
[0989] HATU (193 mg, 0.506 mmol) was added to a stirred solution of
an HCl salt of
(1R,3S,5R)-3-(5-(6((2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)--
1H-imidazol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hexane (Intermediate 124) (100 mg, 0.169 mmol) and
(R)-2-(methoxycarbonylamino)-3-methylbutanoic acid (89 mg, 0.51
mmol) in DMF (1.5 mL) and DIPEA (0.24 mL, 1.4 mmol) and the clear
orange solution was stirred at rt for 3 h. The reaction was diluted
with methanol, filtered and purified by preparative HPLC
(MeOH/water with an ammonium acetate buffer) to yield methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethy-
nyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate (71 mg) as a light yellow solid. LC-MS
retention time 3.706 min; m/z 761.22 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 5% MeOH/95% H.sub.2O/10 mM ammonium acetate and
Solvent B was 5% H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data
was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.15 (s,
1H), 7.96 (s, 1H), 7.80-7.88 (m, 3H), 7.51 (dd, J=8.4, 1.6 Hz, 1H),
7.46 (s, 1H), 7.27 (s, 1H), 5.18 (dd, J=9.0, 4.8 Hz, 1H), 5.13 (dd,
J=8.8, 4.3 Hz, 1H), 4.59 (dd, J=11.8, 6.8 Hz, 2H), 3.68-3.72 (m,
1H), 3.67 (s, 3H), 3.67 (s, 3H), 3.59-3.64 (m, 1H), 2.31-2.60 (m,
4H), 2.08-2.22 (m, 2H), 1.99-2.08 (m, 2H), 1.13 (ddd, J=8.6, 5.8,
5.5 Hz, 2H), 1.00 (dd, J=12.9, 6.9 Hz, 6H), 0.93 (dd, J=6.7, 2.9
Hz, 6H), 0.79 (br. s., 2H).
##STR00674##
Example 75
Mixture of Diastereomers
Methyl
(2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-(((methoxycarbonyl)amino)(-
tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-
-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0990] HATU (110 mg, 0.289 mmol) was added to a stirred solution of
an HCl salt of
(1R,3S,5R)-3-(5-(6-((2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-
-1H-imidazol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[-
3.1.0]hexane (Intermediate 124) (57 mg, 0.096 mmol) and racemic
2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid
(62.7 mg, 0.289 mmol) in DMF (1.0 mL) and DIPEA (0.134 mL, 0.770
mmol) and the clear orange solution was stirred at rt for 3 h. The
reaction was diluted with methanol, filtered and purified by
preparative HPLC (MeOH/water with an ammonium acetate buffer) to
yield methyl
(2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-(((methoxycarbonyl)amino)(tetrah-
ydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)e-
thynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-
-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (37.3 mg) as a yellow
solid and as a mixture of diastereomers. LC-MS retention time 1.522
min; m/z 845.36 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
Mixture of diastereomers. .sup.1H NMR (400 MHz, MeOD) .delta. ppm
8.15 (d, J=12.8 Hz, 1H), 7.96 (s, 1H), 7.80-7.90 (m, 3H), 7.51 (dd,
J=8.5, 1.5 Hz, 1H), 7.46 (d, J=5.5 Hz, 1H), 7.27 (d, J=5.3 Hz, 1H),
5.08-5.24 (m, 2H), 4.61-4.69 (m, 1H), 4.57 (dd, J=8.3, 4.5 Hz, 1H),
3.85-4.03 (m, 5H), 3.63-3.79 (m, 8H), 3.34-3.49 (m, 5H), 2.30-2.63
(m, 4H), 1.99-2.16 (m, 3H), 1.70-1.80 (m, 1H), 1.35-1.66 (m, 6H),
1.16-1.26 (m, 1H), 1.07-1.16 (m, 1H), 0.70-0.83 (m, 2H).
##STR00675##
Example 76
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-((4-chloro-2-((1R,3S,5R)-2-((-
2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3--
yl)-1H-imidazol-5-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[0991] N-Chlorosuccinimide (13.3 mg, 0.100 mmol) was added to a
stirred solution of
((1S)-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)-
amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)eth-
ynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)--
2-methylpropyl)carbamate (38 mg, 0.050 mmol) in DMF (1 mL) and then
the reaction vessel was flushed with nitrogen, sealed and heated at
50.degree. C. for 16 h. The reaction was cooled to rt and
additional N-chlorosuccinimide (7.4 mg) was added. The reaction was
reheated at 50.degree. C. for 3 h, cooled to rt, diluted with MeOH
(.about.0.5 mL), filtered and purified by preparative HPLC
(MeOH/water, TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-((4-chloro-2-((1R,3S,5R)-2-((2S)-2--
((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-
-imidazol-5-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)carbonyl)-2-methylpropyl)carbamate (5.0 mg) as a yellow
solid. LC-MS retention time 4.153 min; m/z 831.02 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 1 mL/min, a gradient of
100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.19 (s, 1H), 8.07 (s, 1H), 7.87-7.99 (m, 3H), 7.58
(dd, J=8.5, 1.3 Hz, 1H), 5.07 (t, J=7.2 Hz, 1H), 4.98-5.03 (m, 1H),
4.54-4.59 (m, 2H), 3.67 (s, 6H), 3.62-3.75 (m, 2H), 2.45-2.50 (m,
2H), 2.41 (dd, J=7.3, 3.5 Hz, 2H), 2.09-2.21 (m, 2H), 1.96-2.08 (m,
2H), 1.08-1.16 (m, 2H), 1.02 (t, J=6.1 Hz, 6H), 0.92-0.97 (m, 6H),
0.75-0.85 (m, 2H).
##STR00676##
Example 77
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(((2R,4r,6S)-2,6-d-
imethyltetrahydro-2H-pyran-4-yl)((methoxycarbonyl)amino)acetyl)-2-azabicyc-
lo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)--
2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[0992] HATU (25.4 mg, 0.067 mmol) was added to a stirred solution
of an HCl salt of methyl
(S)-1-((1R,3S,5R)-3-(5-(4-(6-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl-
)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[-
3.1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (Intermediate
128) (42 mg, 0.061 mmol),
2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylami-
no)acetic acid (Cap-179 stereoisomer 2) (14.9 mg, 0.061 mmol) and
DIEA (0.053 mL, 0.30 mmol) in DMF (2 mL) and the resulting
yellowish solution was stirred at rt for 2 h. The reaction was
concentrated under reduced pressure and the residue was redissolved
into methanol and purified by preparative HPLC. (Solvent A: 05%
MeCN/95% water/10 mM NH.sub.4Ac; Solvent B: 95% MeCN/5% water/10 mM
NH.sub.4Ac; Column: Sunfire Prep MS C18 30.times.100 mm S10;
Wavelength: 220 nM; Flow rate: 40 ml/min; Gradient: 0% B to 80% B
over 30 min with a 5 min. hold time) to yield methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(((2R,4r,6S)-2,6--
dimethyltetrahydro-2H-pyran-4-yl)((methoxycarbonyl)amino)acetyl)-2-azabicy-
clo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-
-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate (26
mg) as a white solid. LC-MS [M+H].sup.+=883; Rt=2.99 min is
product. Column Luna 3u C18 2.times.50 mm; start % B: 0, final % B:
100 Solvent A: 5% ACN/95% H.sub.2O+10 mM Ammonium Acetate; Solvent
B: 95% ACN/5% H.sub.2O+10 mM Ammonium Acetate; flow rate 4 ml/min.
Run time: 5 min. Purity=97%. .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 11.85 (1H, br. s.), 11.73 (1H, br. s.), 8.14-8.27 (2H,
m), 7.90-7.98 (3H, m), 7.82-7.89 (3H, m), 7.79 (2H, d, J=8.24 Hz),
7.64 (1H, s), 7.57 (1H, s), 7.19 (2H, t, J=9.31 Hz), 5.03-5.13 (2H,
m), 4.51 (1H, t, J=7.93 Hz), 4.44 (1H, t, J=7.78 Hz), 3.65 (1H, br.
s.), 3.51-3.60 (7H, m), 3.29-3.42 (7H, m), 2.35-2.47 (2H, m),
2.20-2.33 (2H, m), 2.01-2.11 (2H, m), 1.83-1.92 (3H, m), 1.63 (1H,
d, J=12.21 Hz), 1.57 (1H, d, J=11.60 Hz), 0.96-1.09 (12H, m), 0.89
(4H, d, J=6.71 Hz), 0.72 (2 H, br. s.).
##STR00677##
Example 78
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6'-(2-((1R,3S,5R)-2-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4--
yl)-2,2'-binaphthalen-6-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-
carbonyl)-2-methylpropyl)carbamate
[0993] HATU (91 mg, 0.240 mmol) was added to a stirred solution of
6,6'-bis(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-4-yl)-2-
,2'-binaphthyl (Intermediate 133) (43.9 mg, 0.080 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (42.0 mg, 0.240
mmol) in DMF (1 mL) and DIPEA (0.14 mL, 0.80 mmol) and the clear
brown solution was stirred for 2 h. The reaction mixture was
diluted with MeOH and purified by preparative HPLC (MeOH/water with
0.1% TFA) purified by to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6'-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)-
amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2,-
2'-binaphthalen-6-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbon-
yl)-2-methylpropyl)carbamate (28.9 mg) as a yellow solid. LC-MS
retention time 1.788 min; m/z 863.46 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 1 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.37 (s,
2H), 8.29 (d, J=1.0 Hz, 2H), 8.15 (d, J=8.8 Hz, 2H), 8.06-8.13 (m,
4H), 7.97 (s, 2H), 7.86 (dd, J=8.7, 1.6 Hz, 2H), 5.17 (dd, J=9.3,
7.0 Hz, 2H), 4.57 (d, J=6.8 Hz, 2H), 3.80-3.86 (m, 2H), 3.68 (s,
6H), 2.71 (dd, J=13.7, 9.4 Hz, 2H), 2.46-2.56 (m, 2H), 2.15-2.25
(m, 2H), 2.09 (d, J=8.0 Hz, 2H), 1.08-1.16 (m, 2H), 1.03 (d, J=6.8
Hz, 6H), 0.92-0.96 (m, 6H), 0.88-0.96 (m, 2H).
##STR00678##
Example 79
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,5R)-2-((2S)-2-((2R,4r,6S)-2-
,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-a-
zabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-
-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[0994] HATU (35.4 mg, 0.093 mmol) was added to a stirred solution
of a TFA salt of methyl
(2S)-1-((1R,3S,5R)-3-(5-(6-(4-(2-((1R,5R)-2-azabicyclo[3.1.0]hexan-3-yl)--
1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (47 mg, 0.072
mmol) and
(S)-2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbony-
lamino)acetic acid (23 mg, 0.093 mmol) in DMF (1 mL) and DIPEA
(0.050 mL, 0.29 mmol) and the reaction was stirred for 3 h. The
reaction mixture was diluted with MeOH and purified by preparative
HPLC (MeOH/water with 0.1% TFA) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,5R)-2-((2S)-2-((2R,4r,6S)-2,6-dim-
ethyltetrahydro-2H-pyran-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-azabicy-
clo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-
-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
(31.3 mg) as a yellow solid. LC-MS retention time 1.670 min; m/z
883.43 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.28 (d, J=1.0 Hz, 1H),
8.26 (s, 1H), 8.12 (d, J=8.8 Hz, 1H), 8.07 (d, J=8.5 Hz, 1H),
7.93-7.99 (m, 4H), 7.82-7.92 (m, 4H), 5.15 (td, J=9.6, 6.9 Hz, 2H),
4.58 (t, J=7.4 Hz, 2H), 3.79-3.86 (m, 2H), 3.68 (s, 6H), 3.43-3.53
(m, 2H), 2.65-2.75 (m, 2H), 2.50 (ddd, J=13.6, 6.7, 6.5 Hz, 2H),
2.04-2.24 (m, 4H), 1.54-1.62 (m, 1H), 1.45-1.53 (m, 1H), 0.84-1.21
(m, 18H).
##STR00679##
Example 80
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)c-
arbonyl)-2-methylpropyl)carbamate
[0995] HATU (64.1 mg, 0.169 mmol) was added to a solution of an HCl
salt of
2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-6-(4-(-
2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)quinoli-
ne (Intermediate 139) (50 mg, 0.073 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (29.5 mg, 0.169
mmol) in DMF (1 mL) and DIPEA (0.115 mL, 0.660 mmol) and the
reaction mixture was stirred at rt for 16 h. The reaction was
diluted with MeOH, filtered and purified by preparative HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)p-
henyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbony-
l)-2-methylpropyl)carbamate (55.3 mg) as a yellow solid. LC-MS
retention time 2.883 min; m/z 814.65 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 1 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00680##
Example 81
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-
-yl)-1H-imidazol-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo-
[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0996] HATU (64.1 mg, 0.169 mmol) was added to a solution of an HCl
salt of
2-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-6-(4-(-
2-((1R,3S,5R)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)phenyl)quinoli-
ne (Intermediate 139) (50 mg, 0.073 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (36.6 mg, 0.169 mmol) in DMF (1 mL) and DIPEA (0.12 mL, 0.66
mmol) and the reaction mixture was stirred at rt for 2 h. The
reaction was diluted with MeOH, filtered and purified by
preparative HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield a
TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0-
]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (64.8
mg) as a yellow solid. LC-MS retention time 2.553 min; m/z 898.70
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 10%
MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00681##
Example 82
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl-
)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0997] HATU (102 mg, 0.269 mmol) was added to a stirred solution of
an HCl salt of
(1R,3S,5R)-3-(5-(6-((2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-
-1H-imidazol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[-
3.1.0]hexane (60 mg, 0.134 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (58.4 mg, 0.269 mmol) in DMF (1.2 mL) and DIPEA (0.19 mL, 1.1
mmol) and the reaction was stirred at rt for 2 h. The reaction was
diluted with methanol and purified by preparative HPLC (MeOH/water,
with an ammonium acetate buffer) to yield methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
x-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (48.9 mg)
as a light yellow solid. LC-MS retention time 2.572 min; m/z 845.65
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.14 (s, 1H), 7.96
(s, 1H), 7.80-7.87 (m, 3H), 7.51 (dd, J=8.5, 1.5 Hz, 1H), 7.45 (s,
1H), 7.27 (s, 1H), 5.16 (dd, J=8.8, 5.0 Hz, 1H), 5.11 (dd, J=8.8,
4.5 Hz, 1H), 4.66 (d, J=7.5 Hz, 1H), 4.63 (d, J=7.8 Hz, 1H),
3.90-3.98 (m, 4H), 3.67 (s, 6H), 3.63-3.76 (m, 2H), 3.33-3.44 (m,
4H), 2.32-2.56 (m, 4H), 1.98-2.10 (m, 4H), 1.35-1.67 (m, 8H),
1.08-1.16 (m, 2H), 0.78 (br. s., 2H).
##STR00682##
Example 83
Methyl
((1S)-2-((1R,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl-
)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo--
1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[0998] HATU (27.8 mg, 0.073 mmol) was added to a solution of methyl
(2S)-1-((1R,3S,5R)-3-(5-(6-(4-(2-((1R,5R)-2-azabicyclo[3.1.0]hexan-3-yl)--
1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.-
1.0]hexan-2-yl)-3-methyl-1-oxobutan-2-ylcarbamate (Intermediate
131) (40 mg, 0.061 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (15.90 mg, 0.073 mmol) were dissolved into DMF (0.5 mL) and
DIPEA (0.053 mL, 0.31 mmol) and the reaction was stirred at rt for
2 h. The reaction was diluted with methanol and purified by
preparative HPLC (MeOH/water with an ammonium acetate buffer) to
yield methyl
((1S)-2-((1R,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tet-
rahydro-2H-pyran-4-yl)ethyl)carbamate (3.5 mg) as a light yellow
solid. LC-MS retention time 2.785 min; m/z 855.74 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.17 (s, 1H), 8.10 (s, 1H), 7.93 (dd, J=8.5, 3.0 Hz,
2H), 7.77-7.86 (m, 6H), 7.44 (s, 1H), 7.35 (s, 1H), 5.20 (dd,
J=8.9, 4.6 Hz, 1H), 5.15 (dd, J=8.8, 5.0 Hz, 1H), 4.66 (d, J=7.8
Hz, 1H), 4.61 (d, J=6.8 Hz, 1H), 3.90-3.99 (m, 2H), 3.68 (s, 6H),
3.64-3.76 (m, 3H), 3.34-3.43 (m, 3H), 2.39-2.61 (m, 4H), 2.12-2.23
(m, 1H), 1.99-2.10 (m, 2H), 1.52-1.67 (m, 2H), 1.41-1.51 (m, 1H),
1.08-1.18 (m, 2H), 1.02 (d, J=6.8 Hz, 3H), 0.94 (d, J=6.8 Hz, 3H),
0.76-0.84 (m, 2H).
##STR00683##
Example 84
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2--
((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex--
3-yl)-1H-imidazol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicycl-
o[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate and
##STR00684##
[0999] Example 85
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-(-
(methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)carbonyl)-2-methylpropyl)carbamate
[1000] N-Chlorosuccinimide (6.01 mg, 0.045 mmol) was added to a
stirred solution of a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)p-
henyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbony-
l)-2-methylpropyl)carbamate (Example 80) (26 mg, 0.022 mmol) in DMF
(0.5 mL) and the reaction mixture was stirred at 50.degree. C. for
16 h. The reaction was diluted with MeOH, filtered and purified by
preparative HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield two
products: 1) A TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R-
)-2-((2S)-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]-
hex-3-yl)-1H-imidazol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabi-
cyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate (12.4 mg)
as yellow solid. LC-MS retention time 3.871 min; m/z 882.55 (MH+).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.53 (d, J=8.8 Hz, 1H),
8.26 (d, J=8.8 Hz, 1H), 8.23 (s, 1H), 8.17 (s, 2H), 7.82-7.93 (m,
4H), 5.16 (dd, J=8.8, 5.3 Hz, 1H), 5.06 (t, J=7.2 Hz, 1H),
4.53-4.63 (m, 2H), 3.68-3.76 (m, 2H), 3.67 (s, 6H), 2.44-2.60 (m,
4H), 2.11-2.24 (m, 2H), 1.97-2.09 (m, 2H), 1.09-1.18 (m, 2H), 1.03
(dd, J=6.8, 3.3 Hz, 6H), 0.95 (dd, J=6.8, 2.4 Hz, 6H), 0.75-0.87
(m, 2H). 2) A TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-y-
l)carbonyl)-2-methylpropyl)carbamate (4.8 mg) as yellow solid.
LC-MS retention time 2.98 min; m/z 848.60 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.64 (d,
J=8.8 Hz, 1H), 8.34 (s, 1H), 8.32 (s, 1H), 8.25-8.29 (m, 2H), 8.09
(d, J=8.8 Hz, 1H), 7.86-7.96 (m, 4H), 5.18-5.25 (m, 1H), 5.03-5.09
(m, 1H), 4.53-4.62 (m, 2H), 3.64-3.83 (m, 2H), 3.64-3.69 (m, 6H),
2.42-2.70 (m, 4H), 1.97-2.24 (m, 4H), 1.08-1.17 (m, 2H), 1.02 (dd,
J=6.8, 2.4 Hz, 6H), 0.94 (t, J=6.8 Hz, 6H), 0.76-0.91 (m, 2H).
##STR00685##
Example 86
Methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-(-
(2S)-2-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azab-
icyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol--
2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl-
)carbamate
[1001] N-Chlorosuccinimide (6.68 mg, 0.050 mmol) was added to a
solution of a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0-
]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
(Example 81) (31 mg, 0.025 mmol) in DMF (0.5 mL) and the reaction
mixture was stirred at 50.degree. C. for 16 h. The reaction was
diluted with MeOH, filtered and purified by preparative HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-(4-(4-chloro-2-((1R,3S,5R)-2-((2S)-2-
-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo-
[3.1.0]hex-3-yl)-1H-imidazol-5-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)--
2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carba-
mate (12.1 mg) as yellow solid. LC-MS retention time 3.511 min; m/z
484.17 (1/2MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.60 (d, J=8.8 Hz,
1H), 8.27-8.33 (m, 2H), 8.21 (s, 2H), 7.89-7.95 (m, 2H), 7.83-7.88
(m, 2H), 5.15 (dd, J=8.7, 5.6 Hz, 1H), 5.05 (dd, J=8.3, 6.3 Hz,
1H), 4.64 (dd, J=11.3, 7.8 Hz, 2H), 3.87-4.00 (m, 4H), 3.73-3.81
(m, 2H), 3.68 (s, 6H), 3.33-3.48 (m, 4H), 2.41-2.61 (m, 4H),
1.95-2.16 (m, 4H), 1.39-1.70 (m, 8H), 1.06-1.18 (m, 2H), 0.81 (m,
2H).
##STR00686##
Example 87
Methyl
((1S)-1-(((2S,5S)-2-(4-chloro-5-(4-(6-(4-chloro-2-((2S,5S)-1-((2S)--
2-((methoxycarbonyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-i-
midazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl-
)carbonyl)-2-methylpropyl)carbamate
[1002] N-Chlorosuccinimide (18.88 mg, 0.141 mmol) was to a stirred
solution of a TFA salt of methyl
((1S)-1-(((2S,5S)-2-(4-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate (105 mg, 0.129 mmol) in DMF (3.0 mL) and then and the
reaction was flushed with nitrogen, sealed and heated at 50.degree.
C. overnight. By LC-MS a 1.3:1.6:1 of starting material:mono-Cl
(presumably both regioisomers:dichloro). The reaction was
concentrated, dissolved into MeOH, filtered and purified by
preparative HPLC (MeOH/water with a TFA buffer) to yield recovered
starting material (36.2 mg), a mixture of the two
mono-chloroimidazole regioisomers (56.6 mg) (Example 94 and Example
95) and a TFA salt of methyl
((1S)-1-(((2S,5S)-2-(4-chloro-5-(4-(6-(4-chloro-2-((2S,5S)-1-((2S)-2-((me-
thoxycarbonyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazo-
l-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbo-
nyl)-2-methylpropyl)carbamate (20.4 mg) as a yellow solid. LC-MS
retention time 2.482 min; m/z 885.84 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 1 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00687##
Example 88
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(5-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
4-yl)-1-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)car-
bonyl)-2-methylpropyl)carbamate
[1003] HATU (62.8 mg, 0.165 mmol) was added to a stirred solution
of an HCl salt of
(1R,3S,5R)-3-(5-(4-(5-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-im-
idazol-4-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
exane (Intermediate 144) (42.6 mg, 0.066 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (28.9 mg, 0.165
mmol) in DMF (0.8 mL) and DIPEA (0.092 mL, 0.53 mmol) and the
reaction was stirred at rt for 2 h. The reaction was diluted with
MeOH, filtered and purified by preparative HPLC (MeOH/water with
TFA buffer) to yield the a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(5-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)--
1-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-
-2-methylpropyl)carbamate (53.7 mg) as an off-white solid. LC-MS
retention time 2.821 min; m/z 813.88 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.06 (d,
J=8.5 Hz, 1H), 7.98 (d, J=8.5 Hz, 1H), 7.95 (s, 1H), 7.91 (d, J=8.5
Hz, 2H), 7.78 (s, 1H), 7.65-7.73 (m, 4H), 7.59-7.64 (m, 2H), 5.18
(ddd, J=12.5, 9.2, 7.0 Hz, 2H), 4.59 (dd, J=6.4, 3.4 Hz, 2H),
3.80-3.87 (m, 2H), 3.69 (s, 6H), 2.73 (dt, J=13.4, 9.2 Hz, 2H),
2.48-2.59 (m, 2H), 2.18-2.28 (m, 2H), 2.10 (br. s., 2H), 1.08-1.15
(m, 2H), 1.06 (d, J=7.0 Hz, 3H), 1.04 (d, J=7.0 Hz, 3H), 0.99 (d,
J=6.7 Hz, 3H), 0.95 (d, J=6.7 Hz, 3H), 0.89-0.94 (m, 2H).
##STR00688##
Example 89
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(4-(5-(2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-
-yl)-1H-imidazol-4-yl)-1-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1004] HATU (62.8 mg, 0.165 mmol) was added to a stirred solution
of an HCl salt of
(1R,3S,5R)-3-(5-(4-(5-(2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-im-
idazol-4-yl)naphthalen-1-yl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
exane (Intermediate 144) (42.6 mg, 0.066 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (35.9 mg, 0.165 mmol) in DMF (0.8 mL) and DIPEA (0.092 mL,
0.53 mmol) and the reaction was stirred at rt for 4 h. The reaction
was diluted with MeOH, filtered and purified by preparative HPLC
(MeOH/water with ammonium acetate buffer) to yield methyl
((1S)-2-((1R,3S,5R)-3-(4-(4-(5-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)-1-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (42.6
mg, 0.066 mmol) as an off-white solid. LC-MS retention time 1.965
min; m/z 897.59 (MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 5%
MeOH/95% H.sub.2O/10 mM ammonium acetate and Solvent B was 5%
H.sub.2O/95% MeOH/10 mM ammonium acetate. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.23 (d, J=8.2 Hz, 1H),
7.91 (d, J=8.5 Hz, 1H), 7.82 (d, J=8.2 Hz, 2H), 7.52-7.59 (m, 2H),
7.44-7.51 (m, 4H), 7.41 (s, 1H), 7.21 (s, 1H), 5.23 (dd, J=8.9, 4.6
Hz, 1H), 5.18 (dd, J=8.9, 4.9 Hz, 1H), 4.68 (dd, J=7.5, 2.6 Hz,
2H), 3.89-4.01 (m, 4H), 3.70-3.77 (m, 2H), 3.69 (s, 6H), 3.35-3.45
(m, 4H), 2.43-2.63 (m, 4H), 2.00-2.14 (m, 4H), 1.52-1.71 (m, 6H),
1.41-1.52 (m, 2H), 1.10-1.18 (m, 2H), 0.79-0.86 (m, 2H).
##STR00689##
Example 90
Methyl
((1S)-1-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((1R,3S-
,5R)-3-(4-((6-(2-((1R,3S,5R)-2-((2S)-2-((2R,4r,6S)-2,6-dimethyltetrahydro--
2H-pyran-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-azabicyclo[3.1.0]hex-3--
yl)-1H-imidazol-4-yl)-2-naphthyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3-
.1.0]hex-2-yl)-2-oxoethyl)carbamate
[1005] An HCl salt of
(1R,3S,5R)-3-(5-(6-((2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imid-
azol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
xane (Intermediate 124) (50 mg, 0.073 mmol),
2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylami-
no)acetic acid (Cap-179 stereoisomer 2) (36.9 mg, 0.151 mmol), HATU
(58.6 mg, 0.154 mmol) and DIEA (0.077 mL, 0.441 mmol) were combined
in DMF (3 mL) and the resulting yellow solution was stirred at rt
for 15 h. The volatiles were removed under reduced pressure and the
residue was redissolved in methanol and purified by preparative
HPLC (Solvent A: 10% MeOH/90% water/0.1% TFA; Solvent B: 90%
MeOH/10% water/0.1% TFA; Column Sunfire Prep MS C18 30.times.100 mm
5u; Wavelength: 220 nM; Flow rate: 40 ml/min; Gradient: 10% B to
80% B over 30 min. with a 5 min hold time) to yield a TFA salt of
methyl
((1S)-1-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((1R,3S,5R)-3-
-(4-((6-(2-((1R,3S,5R)-2-((2S)-2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyr-
an-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-
-imidazol-4-yl)-2-naphthyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]h-
ex-2-yl)-2-oxoethyl)carbamate (43 mg) as a white solid. LC-MS
[M+H].sup.+=902; Rt=1.93 min is product. Column PHENOMENEX.RTM. 10u
C18 3.0.times.50 mm; start % B: 0, final % B: 100 Solvent A: 10%
MeOH/90% H.sub.2O+1% TFA; Solvent B: 90% MeOH/10% H.sub.2O+1% TFA;
flow rate 4 mL/min. Run time: 5 min. Purity=98%. .sup.1H NMR (500
MHz, DMSO-d.sub.6) .delta. ppm 8.38 (1 H, br. s.), 8.14-8.22 (1H,
m), 8.09 (1H, d, J=8.85 Hz), 7.93-8.00 (2H, m), 7.62 (1H, d, J=8.85
Hz), 7.25 (2H, t, J=7.48 Hz), 5.02 (2H, t, J=6.87 Hz), 4.51 (1H,
dd, J=7.78, 6.56 Hz), 4.45 (1H, t, J=7.93 Hz), 3.82 (1H, br. s.),
3.67 (1H, d, J=13.73 Hz), 3.54 (6H, s), 3.31-3.42 (3H, m), 2.33
(2H, br. s.), 1.85-1.97 (2H, m), 1.43-1.52 (1H, m), 1.22-1.31 (1H,
m), 1.03-1.12 (10H, m), 0.89-1.01 (9H, m), 0.82 (1H, br. s.), 0.75
(1H, br. s.).
##STR00690##
Example 91
Methyl
((1S)-1-(((2S,3aS,6aS)-2-(4-(6-(4-(2-((2S,3aS,6aS)-1-((2S)-2-((meth-
oxycarbonyl)amino)-3-methylbutanoyl)octahydrocyclopenta[b]pyrrol-2-yl)-1H--
imidazol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)hexahydrocyclopenta[b]p-
yrrol-1(2H)-yl)carbonyl)-2-methylpropyl)carbamate
[1006] HATU (60.9 mg, 0.160 mmol) was added to a stirred solution
of an HCl salt of
(2S,3aS,6aS)-2-(4-(4-(6-(2-((2S,3aS,6aS)-octahydrocyclopenta[b]pyrrol-2-y-
l)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)octahydrocycl-
openta[b]pyrrole (51 mg, 0.073 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (Intermediate
147) (28.1 mg, 0.160 mmol) and DIPEA (0.076 mL, 0.44 mmol) in DCM
(1.5 mL) and the reaction mixture was stirred at rt for 2 h. The
crude reaction mixture was concentrated to dryness and purified by
preparative HPLC (TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((2S,3aS,6aS)-2-(4-(6-(4-(2-((2S,3aS,6aS)-1-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)octahydrocyclopenta[b]pyrrol-2-yl)-1H-imidaz-
ol-4-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)hexahydrocyclopenta[b]pyrrol--
1(2H)-yl)carbonyl)-2-methylpropyl)carbamate (23.7 mg) as a white
solid. LC-MS retention time 2.462 min; m/z 867.8 (MH-). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 1 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 5% acetonitrile/95% H.sub.2O/10 mM ammonium
acetate and Solvent B was 5% H.sub.2O/95% acetonitrile/10 mM
ammonium acetate. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode.
##STR00691##
Example 92
Methyl
((1S)-2-((2S,3aS,6aS)-2-(4-(4-(6-(2-((2S,3aS,6aS)-1-((2S)-2-((metho-
xycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)hexahydrocyclopenta[b-
]pyrrol-2(2H)-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)he-
xahydrocyclopenta[b]pyrrol-1(2H)-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)eth-
yl)carbamate
[1007] HATU (62.1 mg, 0.163 mmol) was added to a stirred solution
of an HCl salt of
(2S,3aS,6aS)-2-(4-(4-(6-(2-((2S,3aS,6aS)-octahydrocyclopenta[b]pyrrol-2-y-
l)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)octahydrocycl-
openta[b]pyrrole (51 mg, 0.073 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (Intermediate
147) (52 mg, 0.074 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (35.5 mg, 0.163 mmol) and DIPEA (0.078 mL, 0.45 mmol) in DCM
(1.5 mL) and the mixture was stirred at rt for 2 h. The crude
reaction mixture was concentrated to dryness and purified by
preparative HPLC 9TFA buffer) to yield a TFA salt of methyl
((1S)-2-((2S,3aS,6aS)-2-(4-(4-(6-(2-((2S,3aS,6aS)-1-((2S)-2-((methoxycarb-
onyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)hexahydrocyclopenta[b]pyrro-
l-2(2H)-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)hexahydr-
ocyclopenta[b]pyrrol-1(2H)-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)car-
bamate (25.8 mg) as a white solid. LC-MS retention time 2.350 min;
m/z 477.5 (1/2MH+). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and Solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00692##
Example 93
3-Chloro-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((3-chloro-5-methoxy-1-i-
soquinolinyl)carbonyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethyn-
yl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-5--
methoxyisoquinoline
[1008] HATU (48.1 mg, 0.127 mmol) was added to a solution of an HCl
salt of
(1R,3S,5R)-3-(5-(6((2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-im-
idazol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]-
hexane (Intermediate 124) (30 mg, 0.051 mmol) and
3-chloro-5-methoxyisoquinoline-1-carboxylic acid (30.1 mg, 0.127
mmol) in DMF (0.8 mL) and DIPEA (0.071 mL, 0.45 mmol) and stirred
at rt overnight. The reaction was diluted with MeOH, filtered, and
purified by preparative HPLC (MeOH/water with an NH.sub.4OAc
buffer) and repurified by preparative HPLC (MeOH/water with a TFA
buffer) to yield a TFA salt of
3-chloro-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((3-chloro-5-methoxy-1--
isoquinolinyl)carbonyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethy-
nyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-5-
-methoxyisoquinoline (40.9 mg) as a yellow solid. LC-MS retention
time 3.621 min; m/z 887.34 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00693##
Example 94
Methyl
((1S)-1-(((2S,5S)-2-(4-(4-(6-(4-chloro-2-((2S,5S)-1-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-5-
-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl-
)-2-methylpropyl)carbamate and
##STR00694##
[1009] Example 95
Methyl
((1S)-1-(((2S,5S)-2-(4-chloro-5-(4-(6-(2-((2S,5S)-1-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-
-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl-
)-2-methylpropyl)carbamate
[1010] These two compounds were prepared in the reaction that
prepared methyl
((1S)-1-(((2S,5S)-2-(4-chloro-5-(4-(6-(4-chloro-2-((2S,5S)-1-((2S)-
-2-((methoxycarbonyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H--
imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidiny-
l)carbonyl)-2-methylpropyl)carbamate (Example 87) and they were
isolated as a mixture of mono-chloroimidazole regioisomers. The two
regioisomers were separated by SCFC on a CHIRALCEL.RTM. OJ-H column
(30.times.250 mm, 5 .mu.m) using Solvents: 75% CO.sub.2-25%
EtOH/0.1% DEA. The absolute regiochemistry of the two
mono-chloroimidazole regioisomers was not determined so each was
arbitrarily assigned. Example 94: Methyl
((1S)-1-(((2S,5S)-2-(4-(4-(6-(4-chloro-2-((2S,5S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-5-yl)-2-
-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate (18.6 mg) was isolated as a yellow solid.
LC-MS retention time 3.756 min; m/z 851.62 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. Example 95: Methyl
((1S)-1-(((2S,5S)-2-(4-chloro-5-(4-(6-(2-((2S,5S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-
-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate (24.1 mg) was isolated as a yellow solid.
LC-MS retention time 3.770 min; m/z 851.64 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00695##
Example 96
Methyl
((1S)-2-((1R,3S,5R)-3-(4-((6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-y-
l)-2-naphthyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-ox-
o-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1011] 4M HCl (0.370 mL, 1.48 mmol) in dioxane was added to a
solution of (1R,3S,5R)-tert-butyl
3-(5-((6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)ethynyl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(Intermediate 150) (69 mg, 0.074 mmol) in dioxane (2 mL) and slurry
was stirred for 1.5 h. The reaction mixture was then concentrated
under a stream of nitrogen and the intermediate product was treated
with
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (20.9 mg, 0.096 mmol), DMF (1.5 mL), DIPEA (0.078 mL, 0.44
mmol) and finally HATU (36.6 mg, 0.096 mmol). The reaction mixture
was stirred at rt overnight, diluted with MeOH, filtered and
purified by preparative HPLC (MeOH/water with 0.1% TFA) to yield a
TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-((6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-n-
aphthyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(t-
etrahydro-2H-pyran-4-yl)ethyl)carbamate (35.3 mg) as a yellow
solid. LC-MS retention time 2.726 min; m/z 803.74 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.27 (s, 1H), 8.18 (s, 1H), 8.06 (d, J=8.8 Hz, 1H),
7.98-8.02 (m, 2H), 7.88 (dd, J=8.7, 1.6 Hz, 1H), 7.74 (s, 1H), 7.67
(dd, J=8.5, 1.5 Hz, 1H), 5.15 (dd, J=9.3, 7.0 Hz, 1H), 5.09 (dd,
J=9.0, 6.3 Hz, 1H), 4.59 (d, J=7.8 Hz, 1H), 4.56 (d, J=6.5 Hz, 1H),
3.91-4.00 (m, 2H), 3.74-3.84 (m, 2H), 3.67 (s, 6H), 3.34-3.45 (m,
2H), 2.70 (dd, J=13.6, 9.5 Hz, 1H), 2.61 (dd, J=13.8, 8.8 Hz, 1H),
2.40-2.54 (m, 2H), 2.14-2.23 (m, 1H), 2.01-2.13 (m, 3H), 1.56-1.64
(m, 1H), 1.38-1.55 (m, 3H), 1.05-1.14 (m, 2H), 1.01 (d, J=6.8 Hz,
3H), 0.92 (d, J=6.8 Hz, 3H), 0.82-0.92 (m, 2H).
##STR00696##
Example 97
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((2R,4r,6S)--
2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2--
azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidaz-
ol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[1012] 4M HCl (0.34 mL, 1.3 mmol) in dioxane was added to a
solution of (1R,3S,5R)-tert-butyl
3-(5-((6-(2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)naphthalen-2-yl)ethynyl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(Intermediate 150) (62.7 mg, 0.067 mmol) in dioxane (2 mL) the
slurry was stirred for 1.5 h. The reaction was then concentrated
under a stream of nitrogen to dryness and then treated with
2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylami-
no)acetic acid (Cap-179 stereoisomer 2) (21.45 mg, 0.087 mmol), DMF
(1.5 mL), DIPEA (0.071 mL, 0.40 mmol) and finally HATU (33.3 mg,
0.087 mmol). The reaction mixture was stirred at rt overnight
diluted with MeOH, filtered and purified by preparative HPLC
(MeOH/water with 0.1% TFA) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((2R,4r,6S)-2,6-di-
methyltetrahydro-2H-pyran-4-yl)-2-((methoxycarbonyl)amino)acetyl)-2-azabic-
yclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-y-
l)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
(28 mg) as a yellow solid. LC-MS retention time 2.898 min; m/z
831.80 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.27 (s, 1H), 8.17
(s, 1H), 8.05 (d, J=8.8 Hz, 1H), 7.98-8.02 (m, 2H), 7.88 (dd,
J=8.5, 1.8 Hz, 1H), 7.74 (s, 1H), 7.66 (dd, J=8.5, 1.5 Hz, 1H),
5.15 (dd, J=9.2, 6.9 Hz, 1H), 5.10 (dd, J=9.0, 6.3 Hz, 1H), 4.56
(t, J=6.0 Hz, 2H), 3.73-3.85 (m, 2H), 3.67 (s, 6H), 3.43-3.54 (m,
2H), 2.70 (dd, J=13.7, 9.4 Hz, 1H), 2.61 (dd, J=13.6, 9.0 Hz, 1H),
2.40-2.54 (m, 2H), 2.01-2.23 (m, 4H), 1.55-1.64 (m, 1H), 1.46 (d,
J=12.0 Hz, 1H), 1.17 (dd, J=6.0, 4.3 Hz, 6H), 1.04-1.13 (m, 3H),
1.01 (d, J=6.8 Hz, 3H), 0.96-1.03 (m, 1H), 0.92 (d, J=6.8 Hz, 3H),
0.88-0.93 (m, 1H), 0.82-0.87 (m, 1H).
##STR00697##
Example 98
Methyl
((1S)-2-((2S,5S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)--
1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-5-methyl-1-pyrroli-
dinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1013] HATU (43.2 mg, 0.114 mmol) was added an HCl salt of
(1R,3S,5R)-3-(4-((6-(2-((2S,5S)-5-methylpyrrolidin-2-yl)-1H-imidazol-4-yl-
)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(Intermediate 154) (27 mg, 0.045 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (24.7 mg, 0.114 mmol) in DMF (1 mL) and DIPEA (0.056 mL, 0.32
mmol) and the reaction mixture was stirred at rt for 2 h. The
reaction was concentrated under a stream of nitrogen, dissolved
into MeOH, filtered and purified by preparative HPLC (MeOH/water
with 0.1% TFA buffer) to yield a TFA salt of methyl
((1S)-2-((2S,5S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imi-
dazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-
-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (31.4 mg) as a
yellow solid. LC-MS retention time 2.658 min; m/z 847.80 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode. The
.sup.1H NMR presents as a .about.1:3.5 mixture of rotamers. The
.sup.1H NMR for the major rotamer is: .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.29 (s, 1H), 8.18 (s, 1H), 8.06 (d, J=8.5 Hz, 1H),
8.01 (d, J=8.8 Hz, 1H), 7.99 (s, 1H), 7.89 (dd, J=8.7, 1.6 Hz, 1H),
7.74 (s, 1H), 7.67 (dd, J=8.4, 1.4 Hz, 1H), 5.19 (dd, J=10.5, 7.0
Hz, 1H), 5.09 (dd, J=8.9, 6.4 Hz, 1H), 4.59 (d, J=7.5 Hz, 1H), 4.22
(d, J=9.3 Hz, 1H), 3.86-4.00 (m, 4H), 3.72-3.82 (m, 2H), 3.67 (s,
6H), 3.23-3.45 (m, 4H), 2.50-2.70 (m, 2H), 2.24-2.50 (m, 3H),
1.90-2.13 (m, 4H), 1.70-1.85 (m, 1H), 1.57 (d, J=6.5 Hz, 3H),
1.30-1.64 (m, 6H), 1.23 (d, J=11.5 Hz, 1H), 1.04-1.12 (m, 1H), 0.85
(br. s., 1H).
##STR00698##
Example 99
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-((6-(2-((2S,5S)-1-((2S)-2-((methoxycarbon-
yl)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-n-
aphthyl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-
-methylpropyl)carbamate
[1014] HATU (43.2 mg, 0.114 mmol) was added to a stirred solution
of an HCl salt of
(1R,3S,5R)-3-(4-((6-(2-((2S,5S)-5-methylpyrrolidin-2-yl)-1H-imidazol-4-yl-
)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(Intermediate 154) (10.1 mg, 0.017 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (7.44 mg, 0.042
mmol) in DMF (0.5 mL) and DIPEA (0.021 mL, 0.119 mmol) and the
reaction mixture was stirred at rt for 2 h. The reaction was
concentrated, dissolved into MeOH, filtered and purified by
preparative HPLC (MeOH/water with 0.1% TFA buffer) to yield a TFA
salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-((6-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthy-
l)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbonyl)-2-methy-
lpropyl)carbamate (9.3 mg) as a yellow solid. LC-MS retention time
2.930 min; m/z 763.75 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR presents as a .about.1:3 mixture of rotamers, the
data for the major rotamer is: .sup.1H NMR (400 MHz, MeOD) .delta.
ppm 8.30 (s, 1H), 8.19 (s, 1H), 8.07 (d, J=8.8 Hz, 1H), 7.99-8.03
(m, 2H), 7.90 (dd, J=8.5, 1.5 Hz, 1H), 7.77 (s, 1H), 7.65-7.71 (m,
1H), 5.19 (dd, J=10.2, 6.9 Hz, 1H), 5.10 (dd, J=9.2, 6.4 Hz, 1H),
4.55 (d, J=6.5 Hz, 1H), 4.13 (d, J=8.8 Hz, 1H), 3.67 (s, 6H),
3.65-3.79 (m, 2H), 1.95-2.68 (m, 8H), 1.57 (d, J=6.5 Hz, 3H), 1.31
(d, J=6.3 Hz, 1H), 1.01 (d, J=7.0 Hz, 3H), 0.98 (d, J=7.0 Hz, 3H),
0.93 (d, J=6.8 Hz, 3H), 0.88 (d, J=6.8 Hz, 3H), 0.84-1.14 (m,
2H).
##STR00699##
Example 100
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl-
)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-
-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1015] 4M HCl (0.32 mL, 1.3 mmol) in dioxane was added to mixture
of a TFA salt of (1R,3S,5R)-tert-butyl
3-(5-(6-((2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)ethynyl)naphthalen-2-yl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(Intermediate 158) (60 mg, 0.064 mmol) in dioxane (0.5 mL) and the
reaction was stirred vigorously for 4 h. The reaction was
concentrated to dryness. Then
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (18.2 mg, 0.084 mmol), DMF (0.5 mL), DIPEA (0.067 mL, 0.39
mmol) and finally HATU (31.8 mg) were added to the crude material
and the reaction was stirred at rt for 1 h. The reaction was
partially concentrated with a stream on nitrogen, diluted with
MeOH, filtered and purified preparative HPLC (MeOH/water with a TFA
buffer) and then repurified preparative HPLC (MeOH/water with an
ammonium acetate buffer). The material was purified a third time by
preparative HPLC (MeOH/water with a TFA buffer) to yield a TFA salt
of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethy-
nyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(t-
etrahydro-2H-pyran-4-yl)ethyl)carbamate (27.7 mg) as a white solid.
LC-MS retention time 3.291 min; m/z 803.67 (MH+). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.27 (s,
1H), 8.19 (s, 1H), 8.06 (d, J=8.8 Hz, 1H), 7.99-8.03 (m, 2H), 7.88
(dd, J=8.5, 1.8 Hz, 1H), 7.78 (s, 1H), 7.68 (dd, J=8.5, 1.5 Hz,
1H), 5.15 (dd, J=9.0, 7.0 Hz, 1H), 5.09 (dd, J=9.3, 6.5 Hz, 1H),
4.80 (d, 2H), 4.54 (d, J=6.5 Hz, 1H), 3.87-3.93 (m, 1H), 3.70-3.79
(m, 3H), 3.68 (s, 3H), 3.67 (s, 3H), 3.54-3.63 (m, 1H), 3.34-3.40
(m, 1H), 2.59-2.74 (m, 2H), 2.40-2.55 (m, 2H), 2.00-2.22 (m, 4H),
1.71-1.81 (m, 2H), 1.51-1.63 (m, 2H), 1.05-1.14 (m, 2H), 1.01 (d,
J=6.8 Hz, 3H), 0.93 (d, J=6.8 Hz, 3H), 0.88 (br. s., 2H).
##STR00700##
Example 101
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbo-
nyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl-
)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-
-1-((3S)-tetrahydro-2H-pyran-3-yl)ethyl)carbamate
[1016] 4M HCl (0.3 mL, 1.3 mmol) in dioxane was added to mixture of
a TFA salt of (1R,3S,5R)-tert-butyl
3-(5-(6-((2-((1R,3S,5R)-2-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-
-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imidazol-5-yl)ethynyl)naphthalen-2-yl)--
1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane-2-carboxylate
(Intermediate 158) (60 mg, 0.064 mmol) in dioxane (0.5 mL) and the
reaction was stirred vigorously for 4 h. The reaction was
concentrated to dryness. Then
(S)-2-(methoxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)acetic
acid (Cap-177a) (18.2 mg, 0.084 mmol), DMF (0.5 mL), DIPEA (0.067
mL, 0.39 mmol) and finally HATU (32 mg, 0.084 mmol) was added to
the crude material and the reaction was stirred at rt for 1 h. The
reaction was partially concentrated, diluted with MeOH, filtered
and purified by preparative HPLC (MeOH/water with a TFA buffer) and
repurified in one injection by Prep HPLC (MeOH/water with an
ammonium acetate buffer) to yield methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)a-
mino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethy-
nyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-((-
3S)-tetrahydro-2H-pyran-3-yl)ethyl)carbamate (30.5 mg) as a white
solid. LC-MS retention time 3.376 min; m/z 803.66 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.14 (s, 1H), 7.94 (s, 1H), 7.78-7.86 (m, 3H), 7.49
(dd, J=8.5, 1.5 Hz, 1H), 7.45 (s, 1H), 7.26 (s, 1H), 5.15 (ddd,
J=12.9, 8.7, 4.5 Hz, 2H), 4.57 (d, J=6.8 Hz, 1H), 3.70-3.86 (m,
3H), 3.67 (br. s., 3H), 3.66 (br. s., 3H), 3.58-3.64 (m, 1H), 3.53
(t, J=8.3 Hz, 1H), 3.45 (dd, J=11.4, 8.2 Hz, 1H), 2.31-2.57 (m,
4H), 1.93-2.17 (m, 5H), 1.67-1.84 (m, 2H), 1.50-1.63 (m, 2H),
1.08-1.17 (m, 2H), 0.98 (d, J=6.8 Hz, 3H), 0.92 (d, J=6.8 Hz, 3H),
0.78 (br. s., 2H).
##STR00701##
Example 102
Methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)pheny-
l)-2-quinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-met-
hylpropyl)carbamate
[1017] HATU (20.99 mg, 0.055 mmol) was added to a solution of an
HCl salt of
2-(2-((2S,4S)-4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)-6-(4-(2-((2S,-
4S)-4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)quinoline
(Intermediate 164) (16.5 mg, 0.024 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (9.7 mg, 0.055
mmol) in DMF (0.5 mL) and DIPEA (0.038 mL, 0.22 mmol) and the
reaction mixture was stirred at rt for 16 h. The reaction was
diluted with MeOH, filtered and purified by preparative HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-2-q-
uinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methylpro-
pyl)carbamate (8.3 mg, 6.94 .mu.mol, 28.9% yield) as a yellow
solid. LC-MS retention time 3.555 min; m/z 818.78 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.60 (d, J=8.5 Hz, 1H), 8.34 (d, J=1.8 Hz, 1H), 8.32
(s, 1H), 8.26-8.31 (m, 1H), 8.21-8.26 (m, 1H), 8.06 (d, J=8.5 Hz,
1H), 7.97-8.03 (m, 1H), 8.00 (d, J=8.5 Hz, 1H), 7.93 (s, 1H),
7.87-7.92 (m, 2H), 5.19-5.36 (m, 2H), 4.29-4.41 (m, 2H), 4.24 (t,
J=6.9 Hz, 2H), 3.67 (s, 6H), 3.44 (q, J=11.0 Hz, 2H), 2.62-2.74 (m,
2H), 2.46-2.61 (m, 2H), 1.97-2.12 (m, 2H), 1.82-1.97 (m, 2H), 1.26
(dd, J=6.3, 3.3 Hz, 6H), 0.91-0.96 (m, 6H), 0.89 (dd, J=6.7, 5.1
Hz, 6H).
##STR00702##
Example 103
Methyl
((1S)-1-(((2S,5S)-2-(4-(6-(4-(2-((2S,5S)-1-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)pheny-
l)-2-quinolinyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-met-
hylpropyl)carbamate
[1018] HATU (15.74 mg, 0.041 mmol) was added to a solution of a
crude HCl salt of
2-(2-((2S,5S)-5-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)-6-(4-(2--
((2S,5S)-5-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)quinoline
(Intermediate 167) (12.35 mg, 0.018 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (7.25 mg, 0.041
mmol) in DMF (0.5 mL) and DIPEA (0.028 mL, 0.16 mmol) and the
reaction mixture was stirred at rt for 16 h. The reaction was
diluted with MeOH, filtered and purified by preparative HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((2S,5S)-2-(4-(6-(4-(2-((2S,5S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-2-q-
uinolinyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methylpro-
pyl)carbamate (3.9 mg) as a yellow solid. LC-MS retention time
3.616 min; m/z 818.73 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.68 (d, J=8.8 Hz,
1H), 8.39 (d, J=1.3 Hz, 1H), 8.30-8.35 (m, 2H), 8.26-8.31 (m, 1H),
8.16 (d, J=8.8 Hz, 1H), 7.99-8.06 (m, 3H), 7.89-7.94 (m, 2H),
5.14-5.31 (m, 2H), 4.14 (dd, J=8.8, 5.0 Hz, 2H), 3.68 (s, 6H),
2.47-2.59 (m, 2H), 2.23-2.43 (m, 4H), 1.95-2.10 (m, 4H), 1.56 (dd,
J=6.7, 3.6 Hz, 6H), 1.01-1.07 (m, 2H), 0.99 (dd, J=6.7, 3.4 Hz,
6H), 0.89 (dd, J=6.8, 1.8 Hz, 6H).
##STR00703##
Example 104
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
4-yl)phenyl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl-
)carbonyl)-2-methylpropyl)carbamate
[1019] HATU (50.7 mg, 0.133 mmol) was added to a solution of an HCl
salt of methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)p-
henyl)-2-quinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-
-methylpropyl)carbamate (Intermediate 173) (41.7 mg, 0.058 mmol)
and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (23.4 mg,
0.133 mmol) in DMF (1 mL) and DIPEA (0.09 mL, 0.5 mmol) and the
reaction mixture was stirred at rt for 16 h. The reaction was
diluted with MeOH, filtered and purified by preparative HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)p-
henyl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)carbo-
nyl)-2-methylpropyl)carbamate (22.3 mg) as a yellow solid. LC-MS
retention time 3.451 min; m/z 815.72 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.41 (s,
1H), 8.43 (d, J=8.5 Hz, 1H), 8.41-8.47 (m, 1H), 8.28 (d, J=1.3 Hz,
2H), 8.03 (d, J=8.5 Hz, 1H), 8.00-8.07 (m, 1H), 7.87-7.95 (m, 3H),
5.10-5.26 (m, 2H), 4.57 (t, J=6.7 Hz, 2H), 3.76-3.89 (m, 2H), 3.68
(s, 6H), 2.70 (dd, J=13.7, 9.2 Hz, 2H), 2.44-2.56 (m, 2H),
2.14-2.26 (m, 2H), 2.04-2.14 (m, 2H), 1.07-1.16 (m, 2H), 1.02 (dd,
J=6.8, 2.0 Hz, 6H), 0.93 (dd, J=6.8, 2.0 Hz, 6H), 0.83-0.99 (m,
2H).
##STR00704##
Example 105
Methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxyca-
rbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-
-yl)-1H-imidazol-4-yl)phenyl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyc-
lo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1020] HATU (12.24 mg, 0.032 mmol) was added to a solution of an
HCl salt of methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycar-
bonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)p-
henyl)-2-quinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-
-methylpropyl)carbamate (Intermediate 173) (10.1 mg, 0.014 mmol)
and (S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (7.0 mg, 0.032 mmol) in DMF (0.5 mL) and DIPEA (0.022 mL, 0.13
mmol) and the reaction mixture was stirred at rt for 2 h. The
reaction was diluted with MeOH, filtered and purified by
preparative HPLC (H.sub.2O-MeOH with 0.1% TFA buffer) to yield a
TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6-(4-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1-
H-imidazol-4-yl)phenyl)-2-quinoxalinyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1-
.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (7.3
mg) as a yellow solid. LC-MS retention time 3.251 min; m/z 899.79
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 9.41 (s, 1H),
8.41-8.47 (m, 2H), 8.27-8.31 (m, 2H), 8.03 (d, J=8.5 Hz, 1H),
8.01-8.06 (m, 1H), 7.87-7.95 (m, 3H), 5.09-5.26 (m, 2H), 4.61 (t,
J=7.2 Hz, 2H), 3.90-4.01 (m, 4H), 3.80-3.90 (m, 2H), 3.68 (s, 6H),
3.34-3.47 (m, 4H), 2.70 (dd, J=13.6, 9.3 Hz, 2H), 2.45-2.57 (m,
2H), 1.99-2.19 (m, 4H), 1.36-1.68 (m, 8H), 1.02-1.16 (m, 2H),
0.79-0.96 (m, 2H).
##STR00705##
Example 106
Methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-((4-chloro-2-((1R,3S,5R)-2-((2-
S)-2-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabic-
yclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-y-
l)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)ca-
rbamate
[1021] N-Chlorosuccinimide (6.97 mg, 0.052 mmol) was added to a
stirred solution of methyl
((1S)-2-((1R,3S,5R)-3-(4-(6((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)am-
ino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-i-
midazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-
-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (Example
82) (21.0 mg, 0.025 mmol) in DMF and then the reaction vessel was
flushed with a stream of nitrogen, sealed and heated at 50.degree.
C. for 3 h. Additional N-chlorosuccinimide (5.0 mg) was added and
the reaction vessel was flushed with nitrogen, sealed and heated at
50.degree. C. for 3 h, then the reaction was cooled and
concentrated under a stream on nitrogen. The remnants were
dissolved into MeOH, filtered, and purified by preparative HPLC
(MeOH/water with TFA buffer) to yield a TFA salt of methyl
((1S)-2-((1R,3S,5R)-3-(4-chloro-5-(6-((4-chloro-2-((1R,3S,5R)-2-((-
2S)-2-((methoxycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabi-
cyclo[3.1.0]hex-3-yl)-1H-imidazol-5-yl)ethynyl)-2-naphthyl)-1H-imidazol-2--
yl)-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)c-
arbamate (8.1 mg) as a yellow solid. LC-MS retention time 4.276
min; m/z 457.47 (1/2MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.18 (s, 1H), 8.07
(s, 1H), 7.98 (d, J=8.8 Hz, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.88 (dd,
J=8.8, 1.8 Hz, 1H), 7.59 (dd, J=8.5, 1.5 Hz, 1H), 5.05 (dd, J=8.5,
6.3 Hz, 1H), 4.99 (t, J=6.9 Hz, 1H), 4.62 (d, J=7.5 Hz, 2H),
3.91-3.98 (m, 4H), 3.74-3.81 (m, 1H), 3.68-3.72 (m, 1H), 3.67 (s,
6H), 3.34-3.45 (m, 4H), 2.45-2.56 (m, 2H), 2.39-2.44 (m, 2H),
1.96-2.11 (m, 4H), 1.37-1.66 (m, 8H), 1.05-1.14 (m, 2H), 0.74-0.84
(m, 2H).
##STR00706##
Example 107
Methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)amino)-2-methylpropyl)-1H-imidazol-4-yl)-2-naphthyl)p-
henyl)-1H-imidazol-2-yl)-2-methylpropyl)carbamoyl)-2-methylpropyl)carbamat-
e
[1022] HATU (77 mg, 0.20 mmol) was added to a stirred solution of
an HCl salt of
(S)-1-(5-(4-(6-(2-((S)-1-amino-2-methylpropyl)-1H-imidazol-4-yl)n-
aphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-methylpropan-1-amine
(Intermediate 178) (57.4 mg, 0.092 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (35.4 mg, 0.202
mmol) and DIPEA (0.096 mL, 0.55 mmol) in DCM (1.5 mL) and the
reaction mixture was stirred at rt for 2 h. The crude reaction
mixture was concentrated to dryness and purified by preparative
HPLC (TFA buffer) to yield a TFA salt of methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl-
)amino)-3-methylbutanoyl)amino)-2-methylpropyl)-1H-imidazol-4-yl)-2-naphth-
yl)phenyl)-1H-imidazol-2-yl)-2-methylpropyl)carbamoyl)-2-methylpropyl)carb-
amate (60.4 mg) as a white solid. LC-MS retention time 2.253 min;
m/z 759.6 (MH-). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and Solvent B was
5% H.sub.2O/95% acetonitrile/10 mM ammonium acetate. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode.
##STR00707##
Example 108
3-Methyl-N-((1S)-2-methyl-1-(4-(4-(6-(2-((1S)-2-methyl-1-((3-methylbutanoy-
l)amino)propyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)prop-
yl)butanamide
[1023] HATU (119 mg, 0.313 mmol) was added to a stirred solution of
an HCl salt of
(S)-1-(5-(4-(6-(2-((S)-1-amino-2-methylpropyl)-1H-imidazol-4-yl)n-
aphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-methylpropan-1-amine
(Intermediate 178) (89 mg, 0.142 mmol), 3-methylbutanoic acid (32
mg, 0.31 mmol) and DIPEA (0.149 mL, 0.855 mmol) in DCM (2 mL) and
the reaction mixture was stirred at rt for 2 h. The crude reaction
mixture was concentrated to dryness and purified twice by
preparative HPLC (TFA buffer) to yield a TFA salt of
3-methyl-N-((1S)-2-methyl-1-(4-(4-(6-(2-((1S)-2-methyl-1-((3-methylbutano-
yl)amino)propyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)pro-
pyl)butanamide (54.6 mg) as a beige solid. LC-MS retention time
2.375 min; m/z 645.6 (MH-). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.30 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 1 mL/min, a gradient of 100% Solvent A/0% Solvent B to
0% Solvent A/100% Solvent B, a gradient time of 2 min, a hold time
of 1 min, and an analysis time of 3 min where Solvent A was 5%
acetonitrile/95% H.sub.2O/10 mM ammonium acetate and
[1024] Solvent B was 5% H.sub.2O/95% acetonitrile/10 mM ammonium
acetate. MS data was determined using a MICROMASS.RTM. Platform for
LC in electrospray mode.
##STR00708##
Example 109
2-((2S,5S)-1-Acetyl-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-1-acetyl--
5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
[1025] HATU (63.2 mg, 0.166 mmol) was added to a solution of
2-((2S,5S)-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 117) (contaminated with ammonium acetate) (38 mg,
0.076 mmol) and
(S)-2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbony-
lamino)acetic acid (40.8 mg, 0.166 mmol) in DMF (1.0 mL) and DIPEA
(0.053 mL, 0.30 mmol) and the reaction was at rt overnight. The
reaction mixture was diluted with MeOH, filtered and purified by
preparative HPLC (MeOH/water with a TFA buffer) to yield a TFA salt
of
2-((2S,5S)-1-acetyl-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-1-acetyl-
-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
e (20 mg) as a yellow solid. LC-MS retention time 3.123 min; m/z
587.62 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.30 (s, 1H), 8.27
(s, 1H), 8.13 (d, J=8.8 Hz, 1H), 8.09 (d, J=8.8 Hz, 1H), 7.93-8.01
(m, 4H), 7.84-7.91 (m, 4H), 5.25 (q, J=8.4 Hz, 2H), 4.31-4.40 (m,
2H), 2.53-2.66 (m, 2H), 2.28-2.40 (m, 4H), 2.25 (s, 3H), 2.25 (s,
3H), 1.88-1.97 (m, 2H), 1.47 (dd, J=6.5, 3.5 Hz, 6H), 1.14-1.19 (m,
1H).
##STR00709##
Example 110
Methyl
(1-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((2S,5S)-2-(-
4-(4-(6-(2-((2S,5S)-1-(((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)((-
methoxycarbonyl)amino)acetyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-
-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-2-oxoethyl)ca-
rbamate
[1026] HATU (61.9 mg, 0.163 mmol) was added to a solution of an HCl
salt of
2-((2S,5S)-5-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,5S)-5-methyl-2-pyr-
rolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 117) (48 mg, 0.074 mmol) and
2-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylami-
no)acetic acid (Cap-179 stereoisomer 2) (39.9 mg, 0.163 mmol) in
DMF (0.8 mL) and DIPEA (0.09 mL, 0.52 mmol) and the reaction was
stirred rt for 2 h. The reaction was diluted with MeOH, filtered
and purified in two injections by prep HPLC (MeOH/water with TFA
buffer) to yield a TFA salt of methyl
(1-((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-((2S,5S)-
-2-(4-(4-(6-(2-((2S,5S)-1-(((2R,4r,6S)-2,6-dimethyltetrahydro-2H-pyran-4-y-
l)((methoxycarbonyl)amino)acetyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-y-
l)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-2-oxoethy-
l)carbamate (55.5 mg) as a light yellow solid. LC-MS retention time
3.506 min; m/z 479.54 (1/2MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. The .sup.1H NMR presented as a complex mixture of
rotamers.
##STR00710##
Example 111
Methyl
((1S)-2-((2S,4S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)--
1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrroli-
dinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1027] HATU (23.99 mg, 0.063 mmol) was added to a solution of an
HCl salt of
(1R,3S,5R)-3-(4-((6-(2-((2S,4S)-4-methylpyrrolidin-2-yl)-1H-imidazol-4-
-yl)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(Intermediate 175) (15 mg, 0.025 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (13.70 mg, 0.063 mmol) in DMF (0.4 mL) and DIPEA (0.03 mL, 0.2
mmol) and the reaction was stirred at rt for 1.5 h. The reaction
was diluted with MeOH, filtered and purified in two injections by
preparative HPLC (MeOH/water with TFA buffer) to a TFA salt of
methyl
((1S)-2-((2S,4S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imi-
dazol-4-yl)ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-
-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (12.0 mg) as a
yellow solid. LC-MS retention time 3.040 min; m/z 847.76 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.28 (s, 1H), 8.18 (s, 1H),
8.05 (d, J=8.8 Hz, 1H), 7.97-8.02 (m, 2H), 7.88 (dd, J=8.8, 1.8 Hz,
1H), 7.76 (s, 1H), 7.67 (dd, J=8.4, 1.4 Hz, 1H), 5.23 (dd, J=10.9,
6.9 Hz, 1H), 5.09 (dd, J=9.0, 6.5 Hz, 1H), 4.59 (d, J=7.8 Hz, 1H),
4.34-4.41 (m, 1H), 4.28 (d, J=8.0 Hz, 1H), 3.86-4.00 (m, 4H),
3.75-3.81 (m, 1H), 3.67 (s, 3H), 3.67 (s, 3H), 3.32-3.50 (m, 5H),
2.39-2.74 (m, 4H), 1.87-2.13 (m, 4H), 1.27-1.65 (m, 8H), 1.25 (d,
J=6.3 Hz, 3H), 1.04-1.14 (m, 1H), 0.86 (br. s., 1H).
##STR00711##
Example 112
Methyl
((1S)-1-(((2S,4S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbon-
yl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-
ethynyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-
-methylpropyl)carbamate
[1028] HATU (60.8 mg, 0.160 mmol) was added to a solution of an HCl
salt of
(1R,3S,5R)-3-(4-((6-(2-((2S,4S)-4-methylpyrrolidin-2-yl)-1H-imidazol-4-
-yl)naphthalen-2-yl)ethynyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(Intermediate 175) (38 mg, 0.064 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (28.0 mg, 0.160
mmol) in DMF (0.7 mL) and DIPEA (0.08 mL, 0.4 mmol) and stirred at
rt for 1.5 h. The reaction was diluted with MeOH, filtered and
purified twice by preparative HPLC (MeOH/water with TFA buffer) to
yield a TFA salt methyl
((1S)-1-(((2S,4S)-2-(4-(6-((2-((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethyny-
l)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methy-
lpropyl)carbamate (19.1 mg) as an off-white solid. LC-MS retention
3.288 time min; m/z 763.74 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of
100%
[1029] Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.28 (s, 1H), 8.18 (s, 1H), 8.05 (d, J=8.8 Hz, 1H),
7.98-8.03 (m, 2H), 7.88 (dd, J=8.7, 1.6 Hz, 1H), 7.76 (s, 1H), 7.67
(dd, J=8.5, 1.5 Hz, 1H), 5.24 (dd, J=10.9, 7.2 Hz, 1H), 5.10 (dd,
J=9.0, 6.5 Hz, 1H), 4.55 (d, J=6.5 Hz, 1H), 4.30-4.37 (m, 1H), 4.23
(d, J=7.3 Hz, 1H), 3.72-3.79 (m, 1H), 3.67 (s, 3H), 3.66 (s, 3H),
3.43 (t, J=10.4 Hz, 1H), 2.49-2.74 (m, 3H), 2.44 (ddd, J=13.6, 6.7,
6.4 Hz, 1H), 2.11-2.22 (m, 1H), 1.98-2.11 (m, 2H), 1.89 (q, J=12.0
Hz, 1H), 1.25 (d, J=6.3 Hz, 3H), 1.06-1.15 (m, 1H), 1.00 (d, J=6.8
Hz, 3H), 0.95-0.98 (m, 1H), 0.94 (d, J=4.0 Hz, 3H), 0.92 (d, J=3.8
Hz, 3H), 0.88 (d, J=6.8 Hz, 3H).
##STR00712##
Example 113
2-((2S,4S)-4-Methyl-1-(3-methylbutanoyl)-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4-
S)-4-methyl-1-(3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-napht-
hyl)phenyl)-1H-imidazole
[1030] HATU (76 mg, 0.200 mmol) was added to a solution of an HCl
salt of
2-((2S,4S)-4-methylpyrrolidin-2-yl)-5-(4-(6-(2-((2S,4S)-4-methylpyrrolidi-
n-2-yl)-1H-imidazol-5-yl)naphthalen-2-yl)phenyl)-1H-imidazole
(Intermediate 196) (64.8 mg, 0.1 mmol), 3-methylbutanoic acid (22.5
mg, 0.220 mmol) and DIPEA (0.122 mL, 0.700 mmol) in DCM (2 mL) and
the mixture was stirred rt overnight. The reaction mixture was
evaporated to dryness and then purified by preparative HPLC to
afford a TFA salt of
2-((2S,4S)-4-methyl-1-(3-methylbutanoyl)-2-pyrrolidinyl)-4-(4-(6-(2-((2S,-
4S)-4-methyl-1-(3-methylbutanoyl)-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naph-
thyl)phenyl)-1H-imidazole (61.7 mg) as an off-white solid. LC-MS
retention time 1.853 min; m/z 671.64 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 1 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00713##
Example 114
Methyl
((1S)-2-((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-im-
idazol-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidiny-
l)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1031] HATU (12.75 mg, 0.034 mmol) was added to a solution of
2-(2-((2S,4S)-4-methylpyrrolidin-2-yl)-1H-imidazol-4-yl)-6-(4-(2-((2S,4S)-
-4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)quinoline
(Intermediate 164) (10 mg, 0.015 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (7.28 mg, 0.034 mmol) in DMF (0.5 mL) and DIPEA (0.023 mL,
0.13 mmol) and the mixture was stirred at rt for 16 h. The reaction
was concentrated and the residue was purified by preparative HPLC
(H.sub.2O-MeOH with 0.1% TFA buffer) to yield a TFA salt of methyl
((1S)-2-((2S,4S)-2-(4-(6-(4-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-
-4-yl)phenyl)-2-quinolinyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-o-
xo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (8.6 mg) as a yellow
solid. LC-MS retention time 3.243 min; m/z 902.90 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped
with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min, and an analysis time
of 5 min where Solvent A was 10% MeOH/90% H.sub.2O/0.1%
trifluoroacetic acid and Solvent B was 10% H.sub.2O/90% MeOH/0.1%
trifluoroacetic acid. MS data was determined using a MICROMASS.RTM.
Platform for LC in electrospray mode. .sup.1H NMR (400 MHz, MeOD)
.delta. ppm 8.61 (d, J=8.8 Hz, 1H), 8.35 (d, J=1.8 Hz, 1H),
8.22-8.33 (m, 3H), 8.07 (d, J=8.8 Hz, 1H), 8.00 (d, J=8.5 Hz, 1H),
7.97-8.04 (m, 1H), 7.89 (d, J=8.5 Hz, 1H), 7.85-7.95 (m, 2H),
5.17-5.33 (m, 2H), 4.32-4.45 (m, 2H), 4.28 (dd, J=8.2, 5.4 Hz, 2H),
3.83-3.98 (m, 4H), 3.67 (s, 6H), 3.22-3.54 (m, 6H), 2.63-2.75 (m,
2H), 2.46-2.61 (m, 2H), 1.83-2.04 (m, 4H), 1.52-1.64 (m, 2H),
1.30-1.53 (m, 6H), 1.26 (dd, J=6.3, 4.0 Hz, 6H).
##STR00714##
Example 115
Methyl
((1S)-1-(((3R)-3-(4-(4-(6-(2-((3R)-4-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-3-morpholinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1-
H-imidazol-2-yl)-4-morpholinyl)carbonyl)-2-methylpropyl)carbamate
[1032] HATU (96 mg, 0.252 mmol) was added to a solution of an HCl
salt of
(R)-3-(5-(4-(6-(2-((R)-morpholin-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)p-
henyl)-1H-imidazol-2-yl)morpholine (Intermediate 181) (71.6 mg,
0.110 mmol) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid
(44.2 mg, 0.252 mmol) in DMF (1.0 mL) and DIPEA (0.13 mL, 0.77
mmol) and the reaction mixture was flushed with nitrogen, sealed
and stirred at rt for 3 h. The reaction was concentrated under a
stream of nitrogen overnight, diluted with MeOH (.about.5 mL),
filtered and purified by preparative HPLC (MeOH/water with a TFA
buffer) to yield a TFA salt of methyl
((1S)-1-(((3R)-3-(4-(4-(6-(2-((3R)-4-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-3-morpholinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imid-
azol-2-yl)-4-morpholinyl)carbonyl)-2-methylpropyl)carbamate (75.5
mg) as a off-white solid LC-MS retention time 3.351 min; m/z 822.07
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR presents as a mixture of rotamers. .sup.1H NMR
(400 MHz, MeOD) .delta. ppm 8.47 (s, 0.5H), 8.33 (s, 0.5H), 8.27
(s, 1H), 7.85-8.17 (m, 10H), 5.95 (d, J=4.5 Hz, 2H), 4.71 (dd,
J=12.5, 7.8 Hz, 1H), 4.35-4.58 (m, 4H), 3.94-4.20 (m, 5H), 3.79 (s,
3H), 3.68 (s, 3H), 3.53-3.78 (m, 3H), 2.76-2.91 (m, 1H), 2.12 (br.
s., 2H), 1.05-1.19 (m, 6H), 0.81-1.00 (m, 6H).
##STR00715##
Example 116
Methyl
((1S)-2-((3R)-3-(4-(4-(6-(2-((3R)-4-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-3-morpholinyl)-1H-imidazol-4-yl)-2-n-
aphthyl)phenyl)-1H-imidazol-2-yl)-4-morpholinyl)-2-oxo-1-(tetrahydro-2H-py-
ran-4-yl)ethyl)carbamate
[1033] HATU (88 mg, 0.231 mmol) was added to a solution of an HCl
salt of
(R)-3-(5-(4-(6-(2-((R)-morpholin-3-yl)-1H-imidazol-4-yl)naphthalen-2-yl)p-
henyl)-1H-imidazol-2-yl)morpholine (Intermediate 181) (65.5 mg,
0.100 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (50.2 mg, 0.231 mmol) in DMF (1.0 mL) and DIPEA (0.12 mL, 0.70
mmol) and the reaction mixture was flushed with nitrogen, sealed
and stirred at rt for 3 h. The reaction was concentrated under a
stream of nitrogen overnight, diluted with MeOH (.about.5 mL),
filtered and purified by preparative HPLC (MeOH/water with a TFA
buffer) to yield a TFA salt of methyl
((1S)-2-((3R)-3-(4-(4-(6-(2-((3R)-4-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-3-morpholinyl)-1H-imidazol-4-yl)-2--
naphthyl)phenyl)-1H-imidazol-2-yl)-4-morpholinyl)-2-oxo-1-(tetrahydro-2H-p-
yran-4-yl)ethyl)carbamate (75 mg) as an off-white solid. LC-MS
retention time 3.120 min; m/z 906.15 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode.
##STR00716##
Example 117
(1R,3S,5R)-2-(3-Methylbutanoyl)-3-(4-(6-((2-((1R,3S,5R)-2-(3-methylbutanoy-
l)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H-i-
midazol-2-yl)-2-azabicyclo[3.1.0]hexane
[1034] HATU (74.8 mg, 0.197 mmol) was added to a stirred solution
of an HCl salt of
(1R,3S,5R)-3-(5-(6-((2-((1R,3S,5R)-2-azabicyclo[3.1.0]hexan-3-yl)-1H-imid-
azol-4-yl)ethynyl)naphthalen-2-yl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]he-
xane (Intermediate 124) (64.7 mg, 0.098 mmol), 3-methylbutanoic
acid (22.09 mg, 0.216 mmol) and DIPEA (0.103 mL, 0.590 mmol) in DCM
(1.5 mL) and the reaction mixture was stirred at rt for 2-3 h. The
reaction was concentrated to dryness and purified by preparative
HPLC (TFA buffer) to yield a TFA salt of
(1R,3S,5R)-2-(3-methylbutanoyl)-3-(4-(6-((2-((1R,3S,5R)-2-(3-methylbutano-
yl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)ethynyl)-2-naphthyl)-1H--
imidazol-2-yl)-2-azabicyclo[3.1.0]hexane (51.5 mg) as a beige
solidified foam. LC-MS retention time 1.775 min; m/z 615.41 (MH+).
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.30 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 1 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min, and an
analysis time of 3 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
##STR00717##
Example 118
Methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-2-piperidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1-
H-imidazol-2-yl)-1-piperidinyl)carbonyl)-2-methylpropyl)carbamate
[1035] HATU (93 mg, 0.245 mmol) was added to a solution of an HCl
salt of
(S)-2-(5-(4-(6-(2-((S)-piperidin-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)p-
henyl)-1H-imidazol-2-yl)piperidine (Intermediate 184) (69.0 mg,
0.106 mmol) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid
(42.9 mg, 0.245 mmol) in DMF (1.0 mL) and DIPEA (0.13 mL, 0.75
mmol) and the reaction was flushed with nitrogen, sealed and
stirred at rt for 1 h. The reaction was concentrated under a stream
of nitrogen overnight, diluted with MeOH (.about.5 mL), filtered
and purified by preparative HPLC (MeOH/water with a TFA buffer) to
yield a TFA salt of methyl
((1S)-1-(((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-3-m-
ethylbutanoyl)-2-piperidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imid-
azol-2-yl)-1-piperidinyl)carbonyl)-2-methylpropyl)carbamate (88.9
mg) as a light yellow solid. LC-MS retention time 3.530 min; m/z
818.08 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0%
[1036] Solvent B to 0% Solvent A/100% Solvent B, a gradient time of
4 min, a hold time of 1 min, and an analysis time of 5 min where
Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and
Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR presents as a mixture of rotamers.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.46 (s, 0.65H), 8.33 (br.
s., 0.35H), 8.27 (s, 1H), 7.84-8.15 (m, 10H), 6.05 (br. s., 1.3H),
5.80-5.90 (m, 0.7H), 4.67-4.78 (m, 1.3H), 4.42-4.50 (m, 2H),
3.99-4.10 (m, 0.7H), 3.78 (br. s., 3.5H), 3.69 (br. s., 2.5H),
3.40-3.52 (m, 0.7H), 2.61-2.73 (m, 1.3H), 2.46-2.59 (m, 1.3H), 2.37
(br. s., 0.7H), 2.03-2.24 (m, 4H), 1.91-2.02 (m, 1.3H), 1.63-1.88
(m, 5.4H), 1.45-1.62 (m, 1.4.H), 1.11 (t, J=6.8 Hz, 7.3H),
0.93-1.03 (m, 4.7H).
##STR00718##
Example 119
Methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-2-piperidinyl)-1H-imidazol-4-yl)-2-n-
aphthyl)phenyl)-1H-imidazol-2-yl)-1-piperidinyl)-2-oxo-1-(tetrahydro-2H-py-
ran-4-yl)ethyl)carbamate
[1037] HATU (84 mg, 0.222 mmol) was added to a solution of an HCl
salt of
(S)-2-(5-(4-(6-(2-((S)-piperidin-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)p-
henyl)-1H-imidazol-2-yl)piperidine (Intermediate 184) (62.5 mg,
0.096 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (48.2 mg, 0.222 mmol) in DMF (1.0 mL) and DIPEA (0.12 mL, 0.68
mmol) and the reaction was flushed with nitrogen, sealed and
stirred at rt for 1 h. The reaction was concentrated under a stream
of nitrogen overnight, diluted with MeOH (.about.5 mL), filtered
and purified by preparative HPLC (MeOH/water with a TFA buffer) to
yield a TFA salt of methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino)-2-(t-
etrahydro-2H-pyran-4-yl)acetyl)-2-piperidinyl)-1H-imidazol-4-yl)-2-naphthy-
l)phenyl)-1H-imidazol-2-yl)-1-piperidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4--
yl)ethyl)carbamate (71.2 mg) as a light yellow solid. LC-MS
retention time 3.348 min; m/z 902.10 (MH+). LC data was recorded on
a Shimadzu LC-10AS liquid chromatograph equipped with a
PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column using a SPD-10AV
UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 0.8 mL/min, a gradient of 100%
Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a gradient
time of 4 min, a hold time of 1 min, and an analysis time of 5 min
where Solvent A was 10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid
and Solvent B was 10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid.
MS data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR presents as a mixture of rotamers.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.45 (s, 0.6H), 8.32 (br.
s., 0.4H), 8.27 (s, 1H), 8.02-8.15 (m, 4.4H), 7.85-8.02 (m, 5.6H),
6.08 (br. s., 1.2H), 5.90 (br. s., 0.8H), 4.70 (br. s., 1.2H),
4.45-4.59 (m, 2H), 3.90-4.10 (m, 4.8H), 3.77 (s, 3.6H), 3.69 (br.
s., 2.4H), 3.36-3.50 (m, 5H), 2.35-2.73 (m, 3H), 1.36-2.22 (m,
20H).
##STR00719##
Example 120
Methyl
((1S)-1-(((2S,5R)-2-(4-(4-(6-(2-((2S,5R)-1-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methy-
lpropyl)carbamate
[1038] HATU (62.0 mg, 0.163 mmol) was added to a solution of an HCl
salt of
2-((2S,5R)-5-methylpyrrolidin-2-yl)-5-(4-(6-(2-((2S,5R)-5-methylpyrrol-
idin-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazole
(Intermediate 187) (46 mg, 0.071 mmol) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (28.6 mg, 0.163
mmol) DMF (0.7 mL) and DIPEA (0.09 mL, 0.5 mmol) and the reaction
vessel was flushed with nitrogen, sealed and stirred at rt for 1 h.
The reaction was concentrated under a stream of nitrogen overnight,
diluted with MeOH (.about.3 mL), filtered and purified by
preparative HPLC (MeOH/water with a TFA buffer) to yield a TFA salt
of methyl
((1S)-1-(((2S,5R)-2-(4-(4-(6-(2-((2S,5R)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-5-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate (53.3 mg) as a light yellow solid. LC-MS retention time
3.255 min; m/z 818.08 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a PHENOMENEX.RTM. Luna
3u C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR presents as a complex mixture of rotamers.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.31 (s, 0.5H), 8.26-8.30
(m, 1.5H), 8.13 (d, J=8.8 Hz, 1H), 8.08 (d, J=8.5 Hz, 1H),
7.93-8.00 (m, 4H), 7.83-7.93 (m, 4H), 5.67 (t, J=7.8 Hz, 0.7H),
5.38-5.45 (m, 1.3H), 4.44-4.68 (m, 3.3H), 3.92 (dd, J=8.2, 4.4 Hz,
0.7H), 3.66 (s, 3.5H), 3.41 (s, 1H), 3.35 (s, 1.5H), 2.53-2.92 (m,
3.3H), 1.96-2.30 (m, 6H), 1.78-1.87 (m, 0.7H), 1.43-1.53 (m, 3.5H),
1.33 (dd, J=6.3, 2.5 Hz, 2.5H), 0.98-1.06 (m, 6H), 0.96 (d, J=6.8
Hz, 2.4H), 0.84 (d, J=6.8 Hz, 3.6H).
##STR00720##
Example 121
Methyl
((1S)-2-((2S,5R)-2-(4-(4-(6-(2-((2S,5R)-1-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-pyrrolidinyl)-1H-im-
idazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl)-
-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1039] HATU (55.8 mg, 0.147 mmol) was added to a solution of vial
an HCl salt of
2-((2S,5R)-5-methylpyrrolidin-2-yl)-5-(4-(6-(2-((2S,5R)-5-methylp-
yrrolidin-2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazole
(Intermediate 187) (41.4 mg, 0.064 mmol) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (31.9 mg, 0.147 mmol) in DMF (0.7 mL) and DIPEA (0.08 mL, 0.4
mmol) and the reaction vessel was flushed with nitrogen, sealed and
stirred at rt for 1 h. The reaction was concentrated under a stream
of nitrogen overnight, diluted with MeOH (.about.3 mL), filtered
and purified by preparative HPLC (MeOH/water with a TFA buffer) to
yield a TFA salt of methyl
((1S)-2-((2S,5R)-2-(4-(4-(6-(2-((2S,5R)-1-((2S)-2-((methoxycarbony-
l)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-pyrrolidinyl)-1H-i-
midazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-1-pyrrolidinyl-
)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (27.6 mg) as a
light yellow solid LC-MS retention time 3.135 min; m/z 902.15
(MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a PHENOMENEX.RTM. Luna 3u C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min, and an analysis time of 5 min where Solvent A was
10% MeOH/90% H.sub.2O/0.1% trifluoroacetic acid and Solvent B was
10% H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. Mixture of rotamers by .sup.1H NMR. .sup.1H NMR (400 MHz,
MeOD) .delta. ppm 8.31 (s, 0.5H), 8.25-8.29 (m, 1.5H), 8.13 (d,
J=8.8 Hz, 1H), 8.08 (dd, J=8.8, 2.5 Hz, 1H), 7.93-8.00 (m, 4H),
7.82-7.92 (m, 4H), 5.71 (t, J=7.9 Hz, 0.8H), 5.37-5.45 (m, 1.2H),
4.47-4.69 (m, 3H), 3.89-4.07 (m, 5H), 3.67 (s, 3H), 3.24-3.43 (m,
7H), 2.57-2.92 (m, 3H), 1.96-2.29 (m, 6H), 1.79-1.87 (m, 1H),
1.35-1.67 (m, 11H), 1.33 (dd, J=6.5, 2.5
[1040] Hz, 3H).
##STR00721##
Example 122
Methyl
((1S)-1-(((2S)-2-(4-(6-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-4-methylene-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)--
2-naphthyl)-1H-imidazol-2-yl)-4-methylene-1-pyrrolidinyl)carbonyl)-2-methy-
lpropyl)carbamate
[1041] HATU (102 mg, 0.269 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((S)-4-methylenepyrrolidin-2-yl)-4-(4-(6-(2-((S)-4-methylenepyrrolidin--
2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazole
(Intermediate 189) (86 mg, 0.148 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (47.1 mg, 0.269
mmol), and Hunig's base (0.234 mL, 1.34 mmol) in DMF (6 mL). The
reaction was stirred 4 h at room temperature and purified by
preparative HPLC (MeOH/water with 0.1% TFA) to afford a bis TFA
salt of methyl
((1S)-1-(((2S)-2-(4-(6-(4-(2-((2S)-1-((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)-4-methylene-2-pyrrolidinyl)-1H-imidazol-4-yl)phenyl)-
-2-naphthyl)-1H-imidazol-2-yl)-4-methylene-1-pyrrolidinyl)carbonyl)-2-meth-
ylpropyl)carbamate (65 mg, 46%). LC-MS retention time 3.22 min;
calcd. for C.sub.46H.sub.53N.sub.8O.sub.6: 813.40 Found m/z 813.7
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeCN/95%
water/10 mM NH.sub.4OAc and Solvent B was 95% MeCN/5% water/10 mM
NH.sub.4OAc. MS data was determined using a MICROMASS.RTM. Platform
for LC in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD)
.delta. ppm 8.29 (s, 1H), 8.28 (s, 1H), 8.15 (d, J=8.9 Hz, 1H),
8.09 (d, J=8.9 Hz, 1H), 8.0-7.85 (m, 8H), 5.50-5.45 (m, 2H), 5.32
(s, 2H), 5.28 (s, 2H), 4.72 (d, J=11.6 Hz, 2H), 4.60 (dd, J=11.3,
3.4 Hz, 2H), 4.21 (dd, J=7.6, 2.8 Hz, 2H), 3.68 (s, 6H), 2.98-2.92
(m, 2H), 2.10-2.06 (m, 2H), 1.06-1.02 (m, 2H), 0.97-0.94 (m,
12H).
##STR00722##
Example 123
Methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methylene-2-pyrrolidinyl)-1H-imida-
zol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methylene-1-pyrrolidinyl)-
-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1042] HATU (102 mg, 0.269 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((S)-4-methylenepyrrolidin-2-yl)-4-(4-(6-(2-((S)-4-methylenepyrrolidin--
2-yl)-1H-imidazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazole
(Intermediate 189) (86 mg, 0.148 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (58.4 mg, 0.269 mmol), and Hunig's base (0.234 mL, 1.34 mmol)
in DMF (6 mL). The reaction was stirred 5 h at room temperature and
purified by preparative HPLC (MeCN/water with 0.1% TFA) to afford a
bis TFA salt of methyl
((1S)-2-((2S)-2-(4-(4-(6-(2-((2S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methylene-2-pyrrolidinyl)-1H-imid-
azol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methylene-1-pyrrolidinyl-
)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate. LC-MS
retention time 2.81 min; calcd. for C.sub.50H.sub.57N.sub.8O.sub.8:
897.43 Found m/z 897.41 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped a PHENOMENEX.RTM.
Luna C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.29 (s, 2H), 8.4 (d, J=8.8 Hz, 1H), 8.09 (d, J=9.0 Hz, 1H),
8.01-7.85 (m, 8H), 5.50-5.44 (m, 2H), 5.32 (s, 2H), 5.28 (s, 2H),
4.75 (d, J=14.6 Hz, 2H), 4.63 (dd, J=13.3, 4.5 Hz, 2H), 4.27 (dd,
J=8.3, 3.5 Hz, 2H), 3.99-3.91 (m, 4H) 3.69 (s, 6H), 3.41-3.37 (m
4H), 3.0-2.94 (m, 2H), 2.02-1.98 (m, 2H), 1.68-1.65 (m, 2H),
1.59-1.32 (m, 8H).
##STR00723##
Example 124
Methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)amino)ethyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H--
imidazol-2-yl)ethyl)carbamoyl)-2-methylpropyl)carbamate
[1043] HATU (149 mg, 0.393 mmol) was added to a stirred solution of
the tetra HCl salt of
(S)-1-(4-(4-(6-(2-((S)-1-aminoethyl)-1H-imidazol-4-yl)naphthalen-2-yl)phe-
nyl)-1H-imidazol-2-yl)ethanamine (Intermediate 191) (111 mg, 0.148
mmol), (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (47.1 mg,
0.196 mmol), and Hunig's base (0.343 mL, 1.96 mmol) in DMF (8 mL).
The reaction was stirred 4 h at room temperature and purified by
preparative HPLC (MeCN/water with 0.1% TFA) to afford a bis TFA
salt methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-3--
methylbutanoyl)amino)ethyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidaz-
ol-2-yl)ethyl)carbamoyl)-2-methylpropyl)carbamate (91 mg, 47%).
LC-MS retention time 2.96 min; calcd. for
C.sub.40H.sub.49N.sub.8O.sub.6: 737.38 Found m/z 737.45
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeOH/95%
water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.35 (s, 1H), 8.27 (s, 1H), 8.13 (d, J=8.9 Hz, 1H), 8.09 (d,
J=8.9 Hz, 1H), 7.99-7.88 (m, 8H), 5.30-5.27 (m, 2H), 3.99-3.97 (m,
2H), 3.70 (s, 6H), 2.14 (br. s, 2H), 1.78-1.75 (m, 6H), 0.99-0.95
(m, 12H).
##STR00724##
Example 125
Methyl
((1S)-2-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)ami-
no)-2-(tetrahydro-2H-pyran-4-yl)acetyl)amino)ethyl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)ethyl)amino)-2-oxo-1-(tetrahydro-2H-pyran--
4-yl)ethyl)carbamate
[1044] HATU (149 mg, 0.393 mmol) was added to a stirred solution of
the tetra HCl salt of
(S)-1-(4-(4-(6-(2-((S)-1-Aminoethyl)-1H-imidazol-4-yl)naphthalen-2-yl)phe-
nyl)-1H-imidazol-2-yl)ethanamine (Intermediate 191) (83 mg, 0.196
mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (85 mg, 0.393 mmol), and Hunig's base (0.343 mL, 1.96 mmol) in
DMF (8 mL). The reaction was stirred 5 h at room temperature and
purified by preparative HPLC (MeCN/water with 0.1% TFA) to afford a
bis TFA salt of methyl
((1S)-2-(((1S)-1-(4-(4-(6-(2-(1S)-1-(((2S)-2-((methoxycarbonyl)ami-
no)-2-(tetrahydro-2H-pyran-4-yl)acetyl)amino)ethyl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)ethyl)amino)-2-oxo-1-(tetrahydro-2H-pyran--
4-yl)ethyl)carbamate (89 mg, 42%). LC-MS retention time 2.64 min;
calcd. for C.sub.44H.sub.53N.sub.8O.sub.8: 821.40 Found m/z 821.80
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% AcCN/95%
water/10 mM NH.sub.4OAc and Solvent B was 95% AcCN/5% water/10 mM
NH.sub.4OAc. MS data was determined using a MICROMASS.RTM. Platform
for LC in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD)
.delta. ppm 8.34 (s, 1H), 8.2 (s, 1H), 8.13 (d, J=8.6 Hz, 1H), 8.09
(d, J=8.6 Hz, 1H), 7.99-7.87 (m, 8H), 5.29-5.26 (m, 2H), 4.02-4.00
(m, 2H), 3.98-3.93 (m, 4H) 3.71 (s, 6H), 3.41-3.37 (m 4H), 2.02
(br. s, 2H), 1.79-1.76 (m, 6H), 1.62 (d, J=13.0 Hz, 2H), 1.49-1.39
(m, 6H).
##STR00725##
Example 126
Methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)(methyl)amino)ethyl)-1H-imidazol-4-yl)-2-naphthyl)phe-
nyl)-1H-imidazol-2-yl)ethyl)(methyl)carbamoyl)-2-methylpropyl)carbamate
[1045] HATU (84 mg, 0.222 mmol) was added to a stirred solution of
the tetra HCl salt of
(S)--N-Methyl-1-(4-(4-(6-(2-((S)-1-(methylamino)ethyl)-1H-imidazol-4-yl)n-
aphthalen-2-yl)phenyl)-1H-imidazol-2-yl)ethanamine (Intermediate
193) (66 mg, 0.111 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (38.9 mg, 0.222
mmol), and Hunig's base (0.194 mL, 1.11 mmol) in DMF (5 mL). The
reaction was stirred 5 h at room temperature and purified by
preparative HPLC (MeCN/water with 0.1% TFA) to afford a bis TFA
salt of methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)am-
ino)-3-methylbutanoyl)(methyl)amino)ethyl)-1H-imidazol-4-yl)-2-naphthyl)ph-
enyl)-1H-imidazol-2-yl)ethyl)(methyl)carbamoyl)-2-methylpropyl)carbamate.
LC-MS retention time 1.78 min; calcd. for
C.sub.42H.sub.53N.sub.8O.sub.6: 765.41 Found m/z 765.41
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 1 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min and an
analysis time of 3 min where Solvent A was 5% MeOH/95% water/0.1%
TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (TFA salt; rotamers, 500 MHz, MeOD) .delta. ppm
8.44/8.34 (s, 1H), 8.27 (s, 1H), 8.13 (d, J=8.6 Hz, 1H), 8.10-7.88
(m, 9H), 6.17-6.14/5.33-5.30 (m, 2H), 4.64-4.6/4.41-4.39 (m, 2H),
3.84/3.66 (s, 3H), 3.83/3.59 (s, 3H), 3.37 (s, 6H), 2.10-2.05 (m,
2H), 1.90-1.87/1.85-1.82 (m, 6H), 1.15-0.90 (m, 12H).
##STR00726##
Example 127
Methyl
((1S)-2-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)ami-
no)-2-(tetrahydro-2H-pyran-4-yl)acetyl)(methyl)amino)ethyl)-1H-imidazol-4--
yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)(methyl)amino)-2-oxo-1-(tetr-
ahydro-2H-pyran-4-yl)ethyl)carbamate
[1046] HATU (84 mg, 0.222 mmol) was added to a stirred solution of
the tetra HCl salt of
(S)--N-Methyl-1-(4-(4-(6-(2-((S)-1-(methylamino)ethyl)-1H-imidazol-4-yl)n-
aphthalen-2-yl)phenyl)-1H-imidazol-2-yl)ethanamine (Intermediate
193) (66 mg, 0.111 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (48 mg, 0.222 mmol), and Hunig's base (0.194 mL, 1.11 mmol) in
DMF (5 mL). The reaction was stirred 5 h at room temperature and
purified by preparative HPLC (MeCN/water with 0.1% TFA) to afford a
bis TFA salt of Methyl methyl
((1S)-2-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-2--
(tetrahydro-2H-pyran-4-yl)acetyl)(methyl)amino)ethyl)-1H-imidazol-4-yl)-2--
naphthyl)phenyl)-1H-imidazol-2-yl)ethyl)(methyl)amino)-2-oxo-1-(tetrahydro-
-2H-pyran-4-yl)ethyl)carbamate. LC-MS retention time 1.58 min;
calcd. for C.sub.46H.sub.57N.sub.8O.sub.8: 849.43 Found m/z 849.46
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.30 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 1 mL/min,
a gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 2 min, a hold time of 1 min and an
analysis time of 3 min where Solvent A was 5% MeOH/95% water/0.1%
TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (TFA salt, rotamers 500 MHz, MeOD) .delta. ppm
8.45/8.34 (s, 1H), 8.28 (s, 1H), 8.14 (d, J=8.6 Hz, 1H), 8.09 (d,
J=8.3 Hz, 1H), 8.06-7.88 (m, 8H), 6.21-6.18/5.38-5.34 (m, 2H),
4.73-4.70/4.47-4.45 (m, 2H), 4.02-3.94 (m, 4H) 3.84/3.63 (s, 3H),
3.83/3.60 (s, 3H), 3.46-3.38 (m 4H), 3.37/3.35 (s, 6H), 2.02 (br.
s, 2H), 1.91-1.88/1.84-1.81 (m, 6H), 1.68 (d, J=13.0 Hz, 2H),
1.53-1.42 (m, 6H).
##STR00727##
Example 128
Methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1((2S)-2-((methoxycarbonyl-
)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-nap-
hthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methyl-
propyl)carbamate
[1047] HATU (32 mg, 0.085 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((2S,4S)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 196) (25 mg, 0.039 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (14.9 mg, 0.085
mmol), and Hunig's base (0.054 mL, 0.308 mmol) in DMF (1 mL). The
reaction was stirred 1 h at room temperature and partially
concentrated by purge of nitrogen gas. The residue was taken up in
MeOH and purified by preparative HPLC (MeCN/water with 0.1% TFA) to
afford a bis TFA salt of methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1((2S)-2-((methoxycarbonyl)amino-
)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)-
phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropyl-
)carbamate (24.5 mg, 57.8%). LC-MS retention time 3.05 min; calcd.
for C.sub.46H.sub.57N.sub.8O.sub.6: 817.44 Found m/z 817.36
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeOH/95%
water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.29 (s, 1H), 8.27 (s, 1H), 8.13 (d, J=8.6 Hz, 1H), 8.08 (d,
J=8.6 Hz, 1H), 7.99-7.85 (m, 8H), 5.27-5.22 (m, 2H), 4.37-4.33 (m,
2H), 4.25-4.23 (m, 2H), 3.68 (s, 6H), 3.47-3.41 (m, 2H), 2.72-2.68
(m 2H), 2.55 (br. s, 2H), 2.07-2.03 (m, 2H), 1.92-1.87 (m, 2H),
1.27-1.26 (m, 6H), 0.95-0.9 (m, 12H).
##STR00728##
Example 129
Methyl
(2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(((methoxycarbonyl)amino)(tetr-
ahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2--
naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxo-1-(tetra-
hydro-2H-pyran-4-yl)ethyl)carbamate
[1048] HATU (64.5 mg, 0.170 mmol) was added to a stirred solution
of the tetra HCl salt of
2-((2S,4S)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 196) (50 mg, 0.077 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (36.8 mg, 0.170 mmol), and Hunig's base (0.11 mL, 0.617 mmol)
in DMF (1 mL). The reaction was stirred 1 h at room temperature and
partially concentrated by purge of nitrogen gas. The residue was
taken up in MeOH and purified by preparative HPLC (MeCN/water with
0.1% TFA) to afford a bis TFA salt of methyl
(2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(((methoxycarbonyl)amino)(tetrahydro-
-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphth-
yl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxo-1-(tetrahydro--
2H-pyran-4-yl)ethyl)carbamate. LC-MS retention time 2.84 min;
calcd. for C.sub.50H.sub.61N.sub.8O.sub.8: 901.46 Found m/z 901.42
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeOH/95%
water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.29 (s, 1H), 8.28 (s, 1H), 8.14 (d, J=8.6 Hz, 1H), 8.09 (d,
J=8.6 Hz, 1H), 8.00-7.85 (m, 8H), 5.25-5.21 (m, 2H), 4.42-4.39 (m,
2H), 4.30-4.27 (m, 2H), 3.96-3.92 (m, 4H), 3.68 (s, 6H), 3.49-3.35
(m 6H), 2.71-2.38 (m, 2H), 2.58-2.54 (m, 2H), 1.97-1.89 (m, 4H),
1.60 (d, J=13.0 Hz, 2H), 1.48-1.26 (m, 12H).
##STR00729##
Example 130
Methyl
(1-cyclopropyl-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(cyclopropyl((me-
thoxycarbonyl)amino)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-n-
aphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxoethyl)carb-
amate
[1049] HATU (39 mg, 0.102 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((2S,4S)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 196) (30 mg, 0.085 mmol),
(S)-2-cyclopropyl-2-(methoxycarbonylamino)acetic acid (17.62 mg,
0.102 mmol), and Hunig's base (0.065 mL, 0.37 mmol) in DMF (1 mL).
The reaction was stirred 1 h at room temperature and partially
concentrated by purge of nitrogen gas. The residue was taken up in
MeOH and purified by preparative HPLC (MeCN/water with 0.1% TFA) to
afford a bis TFA salt of methyl
(1-cyclopropyl-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(cyclopropyl((m-
ethoxycarbonyl)amino)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2--
naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxoethyl)car-
bamate (24.5 mg, 57.8%). LC-MS retention time 3.08 min; calcd. for
C.sub.46H.sub.53N.sub.8O.sub.6: 813.41 Found m/z 813.45
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeOH/95%
water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.32 (s, 1H), 8.27 (s, 1H), 8.13 (d, J=8.6 Hz, 1H), 8.08 (d,
J=8.6 Hz, 1H), 7.99-7.87 (m, 8H), 5.29-5.24 (m, 2H), 4.30 (br. s,
2H), 3.82-3.80 (m, 2H), 3.68 (s, 6H), 3.43-3.37 (m, 2H), 2.72-2.68
(m 2H), 2.55 (br. s, 2H), 1.91-1.86 (m, 2H), 1.25-1.24 (m, 6H),
1.11 (br. s, 2H), 0.60-0.58 (m, 4H), 0.53-0.51 (m, 2H), 0.04-0.38
(m, 2H).
##STR00730##
Example 131
Methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1((2S)-2-((methoxycarbonyl-
)amino)butanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phe-
nyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)propyl)carbamate
[1050] HATU (45 mg, 0.119 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((2S,4S)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 196) (35 mg, 0.054 mmol),
(S)-2-(methoxycarbonylamino)butanoic acid (19.14 mg, 0.119 mmol),
and Hunig's base (0.075 mL, 0.432 mmol) in DMF (1 mL). The reaction
was stirred 1 h at room temperature and partially concentrated by
purge of nitrogen gas. The residue was taken up in MeOH and
purified by preparative HPLC (MeCN/water with 0.1% TFA) to afford a
bis TFA salt of methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1((2S)-2-((methoxycarbonyl)amino-
)butanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1-
H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)propyl)carbamate
(36.4 mg, 61.7%). LC-MS retention time 2.91 min; calcd. for
C.sub.44H.sub.53N.sub.8O.sub.6: 789.41 Found m/z 789.41
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeOH/95%
water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.30 (s, 1H), 8.28 (s, 1H), 8.14 (d, J=8.6 Hz, 1H), 8.09 (d,
J=8.6 Hz, 1H), 8.00-7.86 (m, 8H), 5.27-5.22 (m, 2H), 4.37 (br. s,
2H), 4.24-4.23 (m, 2H), 3.67 (s, 6H), 3.44-3.39 (m, 2H), 2.72-2.68
(m 2H), 2.58 (br. s, 2H), 1.88-1.84 (m, 2H), 1.47-1.25 (m, 6H),
0.98-0.92 (m, 10H).
##STR00731##
Example 132
Methyl
((1S)-1-cyclobutyl-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-cycl-
obutyl-2-((methoxycarbonyl)amino)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imida-
zol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2--
oxoethyl)carbamate
[1051] HATU (45 mg, 0.119 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((2S,4S)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 196) (35 mg, 0.054 mmol),
(S)-2-cyclobutyl-2-(methoxycarbonylamino)acetic acid (22.23 mg,
0.119 mmol) (prepared in a similar manner as Cap-51 using
commercially available
(S)-2-(tert-butoxycarbonylamino)-2-cyclobutylacetic acid as a
starting material) and Hunig's base (0.075 mL, 0.432 mmol) in DMF
(1 mL). The reaction was stirred 1 h at room temperature and
partially concentrated by purge of nitrogen gas. The residue was
taken up in MeOH and purified by preparative HPLC (MeCN/water with
0.1% TFA) to afford a bis TFA salt of methyl
((1S)-1-cyclobutyl-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1((2S)-2-c-
yclobutyl-2-((methoxycarbonyl)amino)acetyl)-4-methyl-2-pyrrolidinyl)-1H-im-
idazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-
-2-oxoethyl)carbamate (25.6 mg, 43.6%). LC-MS retention time 3.13
min; calcd. for C.sub.48H.sub.57N.sub.8O.sub.6: 841.44 Found m/z
841.48 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped a PHENOMENEX.RTM. Luna C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.30 (s, 1H), 8.28 (s, 1H), 8.14 (d, J=8.9 Hz, 1H), 8.09 (d,
J=8.6 Hz, 1H), 8.00-7.86 (m, 8H), 5.26-5.21 (m, 2H), 4.41-4.37 (m,
4H), 3.68 (s, 6H), 3.48-3.43 (m, 2H), 2.71-2.66 (m 4H), 2.56 (br.
s, 2H), 1.96-1.87 (m, 14H), 1.27-1.26 (m, 6H).
##STR00732##
Example 133
Methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl-
)amino)-2-((2R,4S)-2-methyltetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyr-
rolidinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-
-1-pyrrolidinyl)-1-((2S,4R)-2-methyltetrahydro-2H-pyran-4-yl)-2-oxoethyl)c-
arbamate
[1052] HATU (32 mg, 0.085 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((2S,4S)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 196) (25 mg, 0.039 mmol),
2-(methoxycarbonylamino)-2-((2R,4S)-2-methyltetrahydro-2H-pyran-4-yl)acet-
ic acid (Cap-178, stereoisomer 1) (19.61 mg, 0.085 mmol) and
Hunig's base (0.054 mL, 0.308 mmol) in DMF (1 mL). The reaction was
stirred 1 h at room temperature and partially concentrated by purge
of nitrogen gas. The residue was taken up in MeOH and purified by
preparative HPLC (MeCN/water with 0.1% TFA) to afford a bis TFA
salt of methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amino-
)-2-((2R,4S)-2-methyltetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidi-
nyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyr-
rolidinyl)-1-((2S,4R)-2-methyltetrahydro-2H-pyran-4-yl)-2-oxoethyl)carbama-
te (21.3 mg, 45.2%). LC-MS retention time 2.94 min; calcd. for
C.sub.52H.sub.64N.sub.8O.sub.8: 929.49 Found m/z 929.64
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeOH/95%
water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.32 (s, 1H), 8.28 (s, 1H), 8.15-8.12 (m, 1H), 8.09-8.06 (m,
1H), 8.02-7.94 (m, 5H), 7.90-7.86 (m, 3H), 5.28-5.23 (m, 2H),
4.50-4.48 (m, 2H), 4.44 (br. s, 2H), 3.95-3.91 (m, 2H), 3.71-3.68
(s, 10H), 3.46-3.41 (m, 2H), 2.72-2.67 (m, 2H), 2.55-2.52 (m, 2H),
2.19 (br. s, 2H), 1.97-1.92 (m, 2H), 1.60 (br. s, 2H), 1.48-1.46
(m, 2H), 1.39 (br. s, 2H), 1.30-1.26 (m, 8H), 1.14-1.10 (m,
6H).
##STR00733##
Example 134
Methyl
((1S)-1-(((2S,4R)-2-(4-(4-(6-(2-((2S,4R)-1-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methy-
lpropyl)carbamate
[1053] HATU (65 mg, 0.171 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((2S,4R)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4R)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 201) (50 mg, 0.077 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (30 mg, 0.170
mmol), and Hunig's base (0.108 mL, 0.308 mmol) in DMF (1 mL). The
reaction was stirred 1 h at room temperature and partially
concentrated by purge of nitrogen gas. The residue was taken up in
MeOH and purified by preparative HPLC (MeCN/water with 0.1% TFA) to
afford a bis TFA salt of methyl
((1S)-1-(((2S,4R)-2-(4-(4-(6-(2-((2S,4R)-1-((2S)-2-((methoxycarbonyl)amin-
o)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-naphthyl-
)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-methylpropy-
l)carbamate (60.7 mg, 71%). LC-MS retention time 3.12 min; calcd.
for C.sub.46H.sub.57N.sub.8O.sub.6: 817.44 Found m/z 817.46
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. Luna C18 2.0.times.50 mm
column using a SPD-10AV UV-Vis detector at a detector wave length
of 220 nM. The elution conditions employed a flow rate of 0.8
mL/min, a gradient of 100% Solvent A/0% Solvent B to 0% Solvent
A/100% Solvent B, a gradient time of 4 min, a hold time of 1 min
and an analysis time of 5 min where Solvent A was 5% MeOH/95%
water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS
data was determined using a MICROMASS.RTM. Platform for LC in
electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.30 (s, 1H), 8.28 (s, 1H), 8.14 (d, J=8.6 Hz, 1H), 8.09 (d,
J=8.6 Hz, 1H), 8.00-7.86 (m, 8H), 5.43-5.38 (m, 2H), 4.26-4.24 (m,
2H), 4.09-4.05 (m, 2H), 3.69 (s, 6H), 3.64 (br. s, 2H), 2.76-2.73
(m 2H), 235-2.32 (m, 2H), 2.25-2.22 (m, 2H), 2.11-2.09 (m, 2H),
1.22-1.20 (m, 6H), 0.98-0.93 (m, 12H).
##STR00734##
Example 135
Methyl
((1S)-1-(((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol--
4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)car-
bonyl)-2-methylpropyl)carbamate
[1054] HATU (65 mg, 0.171 mmol) was added to a stirred solution of
the tetra HCl salt of
(1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imid-
azol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hexane
(Intermediate 203) (50 mg, 0.078 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (30 mg, 0.111
mmol), and Hunig's base (0.108 mL, 0.621 mmol) in DMF (1 mL). The
reaction was stirred 1 h at room temperature and partially
concentrated by purge of nitrogen gas. The residue was taken up in
MeOH and purified by preparative HPLC (MeCN/water with 0.1% TFA) to
afford a bis TFA salt of methyl
((1S)-1-(((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxy-
carbonyl)amino)-3-methylbutanoyl)-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidazol-
-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabicyclo[3.1.0]hex-2-yl)ca-
rbonyl)-2-methylpropyl)carbamate (54 mg, 63%). LC-MS retention time
3.15 min; calcd. for C.sub.46H.sub.53N.sub.8O.sub.6: 813.41 Found
m/z 813.45 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped a PHENOMENEX.RTM. Luna C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.33 (s, 1H), 8.26 (s, 1H), 8.12 (d, J=8.6 Hz, 1H), 8.07 (d,
J=8.6 Hz, 1H), 7.98-7.86 (m, 8H), 5.80-5.74 (m, 2H), 4.36-4.34 (m,
2H), 4.02-3.99 (m, 2H), 3.73 (s, 6H), 2.99-2.95 (m, 2H), 2.28-2.25
(m 2H), 2.16 (br. s, 2H), 2.04 (br.s, 2H), 1.24-1.20 (m, 2H), 1.08
(br. s, 2H), 1.02-1.00 (m, 12H).
##STR00735##
Example 136
Methyl
((1S)-1-(((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo[-
3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[1055] HATU (62 mg, 0.164 mmol) was added to a stirred solution of
the tetra HCl salt of
(1S,3S,5S)-5-methyl-3-(4-(4-(6-(2-((1S,3S,5S)-5-methyl-2-azabicyclo[3.1.0-
]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabic-
yclo [3.1.0]hexane (Intermediate 209) (50 mg, 0.074 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (28.7 mg, 0.164
mmol), and Hunig's base (0.104 mL, 0.595 mmol) in DMF (1 mL). The
reaction was stirred 1 h at room temperature and partially
concentrated by purge of nitrogen gas. The residue was taken up in
MeOH and purified by preparative HPLC (MeCN/water with 0.1% TFA) to
afford a bis TFA salt of Methyl
((1S)-1-(((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxy-
carbonyl)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H-
-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo-
[3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate (45 mg, 54%).
LC-MS retention time 3.15 min; calcd. for
C.sub.48H.sub.57N.sub.8O.sub.6: 841.44 Found m/z 841.50
[M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min and an
analysis time of 5 min where Solvent A was 5% MeOH/95% water/0.1%
TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.33 (s,
1H), 8.25 (s, 1H), 8.11 (d, J=8.5 Hz, 1H), 8.06 (d, J=8.7 Hz, 1H),
7.97-7.85 (m, 8H), 5.82-5.75 (m, 2H), 4.31-4.28 (m, 2H), 3.73-3.71
(m, 8H), 2.75-2.69 (m, 2H), 2.44-2.37 (m 2H), 2.17-2.12 (m, 2H),
1.40 (s, 6H), 1.1-1.11 (m, 4H), 1.01-0.99 (m, 12H).
##STR00736##
Example 137
Methyl
((1S)-2-((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxyca-
rbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-azabicyclo[3.-
1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-met-
hyl-2-azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)c-
arbamate
[1056] HATU (62 mg, 0.164 mmol) was added to a stirred solution of
the tetra HCl salt of
(1S,3S,5S)-5-methyl-3-(4-(4-(6-(2-((1S,3S,5S)-5-methyl-2-azabicyclo[3.1.0-
]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-azabic-
yclo [3.1.0]hexane (Intermediate 209) (50 mg, 0.074 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (35.5 mg, 0.164 mmol), and Hunig's base (0.104 mL, 0.595 mmol)
in DMF (1 mL). The reaction was stirred 1 h at room temperature and
partially concentrated by purge of nitrogen gas. The residue was
taken up in MeOH and purified by preparative HPLC (MeCN/water with
0.1% TFA) to afford a bis TFA salt of methyl
((1S)-2-((1S,3S,5S)-3-(4-(4-(6-(2-((1S,3S,5S)-2-((2S)-2-((methoxycarbonyl-
)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-5-methyl-2-azabicyclo[3.1.0]he-
x-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2--
azabicyclo[3.1.0]hex-2-yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbama-
te (46.3 mg, 51%). LC-MS retention time 3.15 min; calcd. for
C.sub.52H.sub.61N.sub.8O.sub.6: 925.46 Found m/z 463.36
[M/2+H].sup.+. LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped a PHENOMENEX.RTM. C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min and an
analysis time of 5 min where Solvent A was 5% MeOH/95% water/0.1%
TFA and Solvent B was 95% MeOH/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.31 (s,
1H), 8.25 (s, 1H), 8.10 (d, J=8.8 Hz, 1H), 8.06 (d, J=8.8 Hz, 1H),
7.97-7.85 (m, 8H), 5.80-5.74 (m, 2H), 4.36-4.35 (m, 2H), 3.99-3.95
(m, 4H), 3.76-3.74 (m, 8H), 3.43-3.35 (m, 4H), 2.75-2.68 (m, 2H),
2.44-2.37 (m 2H), 2.06-2.04 (m, 2H), 1.74 (app. d, J=13 Hz, 2H),
1.52-1.35 (m, 12H), 1.22-1.19 (m, 2H), 1.14-1.12 (m, 2H).
##STR00737##
Example 138
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo[-
3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[1057] HATU (31 mg, 0.082 mmol) was added to a stirred solution of
the tetra HCl salt of
(1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-5-methyl-2-a-
zabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate
(Intermediate 211) (25 mg, 0.037 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (14 mg, 0.082
mmol), and Hunig's base (0.052 mL, 0.297 mmol) in DMF (1 mL). The
reaction was stirred 1 h at room temperature and partially
concentrated by purge of nitrogen gas. The residue was taken up in
MeOH and purified by preparative HPLC (MeCN/water with 0.1% TFA) to
afford a bis TFA salt of Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo[3.1.0]-
hex-2-yl)carbonyl)-2-methylpropyl)carbamate (17 mg, 41%). LC-MS
retention time 3.10 min; calcd. for C.sub.48H.sub.57N.sub.8O.sub.6:
841.44 Found m/z 841.542 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped a PHENOMENEX.RTM.
C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.27 (s, 1H), 8.26 (s, 1H), 8.12 (d, J=8.5 Hz, 1H), 8.07 (d,
J=8.7 Hz, 1H), 7.98-7.93 (m, 4H), 7.88-7.85 (m, 4H), 5.09-5.04 (m,
2H), 4.55-4.53 (m, 2H), 3.38 (s, 6H), 3.57 (br.s, 2H), 2.81-2.75
(m, 2H), 2.33-2.27 (m 2H), 2.22-2.16 (m, 2H), 1.44 (s, 3H), 1.43
(s, 3H), 1.05-0.93 (m, 16H).
##STR00738##
Example 139
Methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxyc-
arbonyl)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H--
imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo[-
3.1.0]hex-2-yl)carbonyl)-2-methylpropyl)carbamate
[1058] HATU (34.8 mg, 0.092 mmol) was added to a stirred solution
of the tetra HCl salt of
(1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-(tert-butoxycarbonyl)-5-methyl-2-a-
zabicyclo[3.1.0]hex-3-yl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-
-2-yl)-5-methyl-2-azabicyclo[3.1.0]hexane-2-carboxylate
(Intermediate 211) (28 mg, 0.042 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (20 mg, 0.092 mmol), and Hunig's base (0.085 mL, 0.333 mmol)
in DMF (2 mL). The reaction was stirred 1 h at room temperature and
partially concentrated by purge of nitrogen gas. The residue was
taken up in MeOH and purified by preparative HPLC (MeCN/water with
0.1% TFA) to afford a bis TFA salt of methyl
((1S)-1-(((1R,3S,5R)-3-(4-(4-(6-(2-((1R,3S,5R)-2-((2S)-2-((methoxycarbony-
l)amino)-3-methylbutanoyl)-5-methyl-2-azabicyclo[3.1.0]hex-3-yl)-1H-imidaz-
ol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-5-methyl-2-azabicyclo[3.1.0]-
hex-2-yl)carbonyl)-2-methylpropyl)carbamate (13 mg, 25%). LC-MS
retention time 2.92 min; calcd. for C.sub.52H.sub.61N.sub.8O.sub.6:
925.46 Found m/z 92.53 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped a PHENOMENEX.RTM.
C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.28 (s, 2H), 8.14 (d, J=8.8 Hz, 1H), 8.07 (d, J=8.6 Hz, 1H),
8.00-7.96 (m, 4H), 7.91-7.84 (m, 4H), 5.08-5.02 (m, 2H), 4.59-4.56
(m, 2H), 4.00-3.94 (m, 4H), 3.69 (s, 6H), 3.62 (br. s, 2H),
3.44-3.38 (m, 4H), 2.81-2.78 (m, 2H), 2.31-2.29 (m, 2H), 2.09 (br.
s, 2H), 1.60-1.47 (m, 8H), 1.43 (s, 3H), 1.44 (s, 3H), 1.04 (br. s,
2H), 0.98-0.97 (m, 2H).
##STR00739##
Example 140
Methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbony-
l)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-i-
midazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl-
)carbonyl)-2-methylpropyl)carbamate
[1059] HATU (97 mg, 0.254 mmol) was added to a stirred solution of
the tetra HCl salt of
2-((2S,4S)-4-methyl-2-pyrrolidinyl)-4-(4-(6-(2-((2S,4S)-4-methyl-2-pyrrol-
idinyl)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazole
(Intermediate 196) (150 mg, 0.231 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (50 mg, 0.231 mmol), and Hunig's base (0.162 mL, 0.925 mmol)
in DMF (1 mL). The reaction was stirred 1 h at room temperature
before addition of (S)-2-(methoxycarbonylamino)-3-methylbutanoic
acid (40.5 mg, 0.231 mmol) and add'l Hunig's base (0.162 mL, 0.925
mmol) and HATU (97 mg, 0.254 mmol). The mixture was stirred further
at rt for 1 h and partially concentrated by purge of nitrogen gas.
The residue was taken up in MeOH and purified by preparative HPLC
(MeCN/water with 0.1% TFA). A second method of chromatography was
applied to separate the isomers: SFC Diol-HL column, 20% methanol
with 0.1% DEA; 4.6.times.250 mm, 5 .mu.m Mobile Phase: 80%
CO.sub.2-20% methanol with 0.1% DEA; Temp: 35.degree. C.; Pressure:
150 bar; Flow rate: 2 ml/min; UV monitored at 328 nm. The isolated
sample was taken up in methanol (3 mL) and neat TFA (50 mL) was
added at rt. The mixture was concentrated down to dryness and
placed on high vacuum to afford a bis TFA salt of methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazo-
l-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbo-
nyl)-2-methylpropyl)carbamate (78.2 mg, 29.5% yield). LC-MS
retention time 2.95 min; calcd. for C.sub.48H.sub.59N.sub.8O.sub.2:
859.45 Found m/z 859.50 [M+H].sup.+. LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped a PHENOMENEX.RTM.
Luna C18 2.0.times.50 mm column using a SPD-10AV UV-Vis detector at
a detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.30 (s, 1H), 8.28 (s, 1H), 8.15 (d, J=8.6 Hz, 1H), 8.09 (d,
J=8.6 Hz, 1H), 8.01-7.85 (m, 8H), 5.26-5.22 (m, 2H), 4.42-4.36 (m,
2H), 4.30-4.27 (m, 1H), 4.24-4.23 (m, 1H), 3.95-3.91 (m, 2H), 3.68
(s, 6H), 3.47-3.36 (m 2H), 3.08-3.04 (m, 1H), 2.70 (br. s, 2H),
2.55 (br. s, 2H), 2.08-2.03 (m, 1H), 1.97-1.87 (m, 3H), 1.59 (d,
J=13.0 Hz, 1H), 1.48-1.26 (m, 10H) 0.95-0.90 (m, 6H).
##STR00740##
Example 141
Methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbonyl-
)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-nap-
hthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxo-1-(tetrahyd-
ro-2H-pyran-4-yl)ethyl)carbamate
[1060] The other isomer as reported in Example 140. LC-MS retention
time 2.98 min; calcd. for C.sub.48H.sub.59N.sub.8O.sub.7: 859.45
Found m/z 859.58 [M+H].sup.+. LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped a PHENOMENEX.RTM. Luna C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.29 (s, 1H), 8.28 (s, 1H), 8.13 (d, J=8.6 Hz, 1H), 8.09 (d,
J=8.6 Hz, 1H), 7.99-7.86 (m, 4H), 7.91-7.85 (m, 4H), 5.26-5.21 (m,
2H), 4.41-4.36 (m, 4H), 4.28 (d, J=8.24 Hz, 1H), 4.24 (d, J=7.32
Hz, 1H), 3.95-3.91 (m, 2H), 3.68 (s, 6H), 3.47-3.36 (m 2H),
2.73-2.66 (m, 2H), 2.59-2.53 (m, 2H), 2.07-2.03 (m, 1H), 1.97-1.87
(m, 3H), 1.59 (d, J=13.0 Hz, 1H), 1.51-1.25 (m, 11H) 0.95-0.90 (m,
6H).
##STR00741##
Example 142
Methyl
((1S)-1-(((2S,4S)-2-(4-chloro-5-(6-(4-(4-chloro-2-((2S,4S)-1-((2S)--
2-((methoxycarbonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-i-
midazol-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl-
)carbonyl)-2-methylpropyl)carbamate
[1061] The NCS (18.0 mg, 0.135 mmol) was added to a nitrogen purged
solution of Example 128 (55 mg, 0.067 mmol) in DMF (1 mL) and the
reaction mixture was stirred for 18 h at 50.degree. C. The solvent
was removed by concentrate, and the crude product was taken up in
CH.sub.2Cl.sub.2 and charged to a 4 g Thompson silica gel
cartridge. Gradient elution was performed from 5-100% B over 1 L.
A/B (A=CH.sub.2Cl.sub.2; B=10% MeOH/EtOAc). A sample of the
purified product was taken up in MeOH and subject to preparative
HPLC (MeCN/water with 0.1% TFA) to afford a bis TFA salt of methyl
((1S)-1-(((2S,4S)-2-(4-chloro-5-(6-(4-(4-chloro-2-((2S,4S)-1-((2S)-2-((me-
thoxycarbonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazo-
l-5-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbo-
nyl)-2-methylpropyl)carbamate. LC-MS retention time 4.48 min;
calcd. for C.sub.46H.sub.55Cl.sub.2N.sub.8O.sub.6: 885.36 Found m/z
885.34 [M+H].sup.+. LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped a PHENOMENEX.RTM. Luna C18
2.0.times.50 mm column using a SPD-10AV UV-Vis detector at a
detector wave length of 220 nM. The elution conditions employed a
flow rate of 0.8 mL/min, a gradient of 100% Solvent A/0% Solvent B
to 0% Solvent A/100% Solvent B, a gradient time of 4 min, a hold
time of 1 min and an analysis time of 5 min where Solvent A was 5%
MeOH/95% water/0.1% TFA and Solvent B was 95% MeOH/5% water/0.1%
TFA. MS data was determined using a MICROMASS.RTM. Platform for LC
in electrospray mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta.
ppm 8.20 (s, 1H), 8.18 (s, 1H), 8.03 (d, J=8.6 Hz, 1H), 8.00 (d,
J=8.6 Hz, 1H), 7.90-7.83 (m, 6H), 5.09-5.04 (m, 2H), 4.30-4.7 (m,
2H), 4.24-4.23 (m, 2H), 3.68 (s, 6H), 3.43-3.99 (m, 2H), 2.58-2.56
(m 2H), 2.43 (br. s, 2H), 2.05-2.04 (m, 2H), 1.93-1.87 (m, 2H),
1.24-1.22 (m, 6H), 0.97-0.91 (m, 12H).
##STR00742##
Example 143
Methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(4-chloro-2-((2S,4S)-1-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-5-
-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl-
)-2-methylpropyl)carbamate
[1062] HATU (34 mg, 0.090 mmol) was added to a stirred solution of
an HCl salt of
4-chloro-2-((2S,4S)-4-methylpyrrolidin-2-yl)-5-(6-(4-(2-((2S,4S)--
4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-1H-imida-
zole (30.0 mg, 0.041 mmol),
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (15.7 mg, 0.090
mmol), and Hunig's base (0.057 mL, 0.33 mmol) in DMF (1 mL). The
reaction was stirred 1 h at room temperature and partially
concentrated by purge of nitrogen gas. The residue was taken up in
MeOH and purified by preparative HPLC (MeCN/water with 0.1% TFA) to
afford a TFA salt of methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(4-chloro-2-((2S,4S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-5-yl)-2-
-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate (24.5 mg) as an off-white solid. LC-MS
retention time 3.77 min; calcd. for
C.sub.46H.sub.56ClN.sub.8O.sub.6: 851.40 Found m/z 851.42 [M+H]. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped a PHENOMENEX.RTM. C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% MeOH/95% water/0.1% TFA and Solvent
B was 95% MeOH/5% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.23 (s, 1H), 8.22 (s, 1H),
8.06 (d, J=8.9 Hz, 1H), 8.02 (d, J=8.9 Hz, 1H), 7.98 (d, J=8.2 Hz,
2H), 7.93-7.89 (m, 3H), 7.85 (d, J=8.9 Hz, 2H), 5.26-5.22 (m, 1H),
5.07-5.04 (m, 1H), 4.36 (t, J=9.1 Hz 1H), 4.29-4.23 (m, 3H), 3.68
(s, 3H), 3.67 (s, 3H), 3.41 (dt, J=10.7, 4.3 Hz, 2H), 2.72-2.67 (m,
1H), 2.58-2.53 (m 2H), 2.45-2.40 (m, 1H), 2.06-2.02 (m, 2H),
1.91-1.83 (m, 2H), 1.26 (d, J=6.1 Hz, 3H), 1.22 (d, J=8.9 Hz, 3H),
0.97-0.91 (m, 12H).
##STR00743##
Example 144
Methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(4-chloro-2-((2S,4S)-1-((2S)-2-((methox-
ycarbonyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidin-
yl)-1H-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrr-
olidinyl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1063] HATU (51 mg, 0.134 mmol) was added to a stirred solution of
an HCl salt of
4-chloro-2-((2S,4S)-4-methylpyrrolidin-2-yl)-5-(6-(4-(2-((2S,4S)--
4-methylpyrrolidin-2-yl)-1H-imidazol-5-yl)phenyl)naphthalen-2-yl)-1H-imida-
zole (32.7 mg, 0.061 mmol),
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (29.2 mg, 0.134 mmol), and Hunig's base (0.085 mL, 0.49 mmol)
in DMF (1 mL). The reaction was stirred 1 h at room temperature and
partially concentrated by purge of nitrogen gas. The residue was
taken up in MeOH and purified by preparative HPLC (MeCN/water with
0.1% TFA) to afford a TFA salt of methyl
((1S)-2-((2S,4S)-2-(4-(4-(6-(4-chloro-2-((2S,4S)-1-((2S)-2-((methoxycarbo-
nyl)amino)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-
-imidazol-5-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidin-
yl)-2-oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (22 mg) as a
light yellow solid. LC-MS retention time 3.55 min; calcd. for
C.sub.50H.sub.60ClN.sub.8O.sub.8: 935.42 Found m/z 935.52 [M+H]. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped a PHENOMENEX.RTM. C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% MeOH/95% water/0.1% TFA and Solvent
B was 95% MeOH/5% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.21 (s, 2H), 8.05 (d, J=8.9
Hz, 1H), 8.02 (d, J=8.9 Hz, 1H), 7.92 (d, J=8.6 Hz, 2H), 7.93-7.89
(m, 3H), 7.85 (d, J=8.2 Hz, 2H), 5.25-5.22 (m, 1H), 5.06-5.02 (m,
1H), 4.40 (t, J=8.9 Hz 1H), 4.31-4.27 (m, 3H), 3.95-3.92 (m, 4H),
3.68 (s, 3H), 3.67 (s, 3H), 3.46-3.36 (m, 6H), 2.72-2.67 (m, 1H),
2.57-2.52 (m 2H), 2.45-2.40 (m, 1H), 1.98-1.86 (m, 4H), 1.61-1.59
(m, 2H), 1.56-131. (m, 6H), 1.27 (d, J=6.4 Hz, 3H), 1.22 (d, J=6.4
Hz, 3H).
##STR00744##
Example 145
Methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(4-chloro-2-((2S,4S)-1-((2S)-2-((metho-
xycarbonyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-5-
-yl)phenyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl-
)-2-methylpropyl)carbamate
[1064] NCS (108 mg, 0.808 mmol) was added to a solution of a TFA
salt of methyl
((1S)-1-(((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-((2S)-2-((methoxycarbon-
yl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2-n-
aphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-meth-
ylpropyl)carbamate (550 mg, 1.385 mmol) in dry DMF (10 mL) and the
mixture was heated at 50.degree. C. for 18 h. The solvent was
removed by rotary evaporation under high vacuum and the residue was
purified utilizing a Thompson 25 g silica gel cartridge (gradient
elution 5%-100% B over 1 L; Solvent B=10% MeOH in ethyl acetate and
Solvent A=dichloromethane). The partially purified product was
subjected to prep HPLC (CH.sub.3CN/H.sub.2O/TFA) on a
Waters-Sunfire column (30.times.100 mm S5) 0-80% B over 35 min and
the mono chloro adducts (116 mg, 20% as a 7:1 mixture favoring
Example 145) were separated from dichloro adduct (see Example 142).
The mono chloro adducts were further purified by chiral SFC
chromatography; CHIRALCEL.RTM. OJ-H column (85% CO.sub.2; 15% EtOH
with 0.1% DEA) to yield methyl
((1S)-1-(((2S,4S)-2-(4-(6-(4-(4-chloro-2-((2S,4S)-1-((2S)-2-((methoxycarb-
onyl)amino)-3-methylbutanoyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-5-yl)ph-
enyl)-2-naphthyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)carbonyl)-2-me-
thylpropyl)carbamate (10 mg) as a yellowish residue. LC-MS
retention time 3.75 min; calcd. for
C.sub.46H.sub.56ClN.sub.8O.sub.6: 851.40 Found m/z 851.39 [M+H]. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% methanol/95% water/0.1% TFA and
Solvent B was 95% methanol/5% water/0.1% TFA. MS data was
determined using a MICROMASS.RTM. Platform for LC in electrospray
mode. .sup.1H NMR (TFA salt, 500 MHz, MeOD) .delta. ppm 8.20 (s,
1H), 8.15 (s, 1H), 8.01 (d, J=8.2 Hz, 1H), 7.98 (d, J=8.6 Hz, 1H),
7.90 (d, J=8.6 Hz, 1H), 7.86 (d, J=7.0 Hz, 1H), 7.83 (d, J=7.3 Hz,
2H), 7.79 (d, J=8.2 Hz, 2H), 7.44 (s, 1H), 5.14-5.10 (m, 1H),
5.06-5.02 (m, 1H), 4.27-4.22 (m, 4H), 3.67 (s, 6H), 3.46-3.47 (m,
2H), 2.57-2.51 (m, 2H), 2.45-2.39 (m 2H), 2.06-2.01 (m, 2H),
1.95-1.90 (m, 2H), 1.23-1.22 (m, 6H), 0.97-0.91 (m, 12H).
##STR00745##
Example 146
Methyl
(2-((2S,4S)-2-(4-(4-(6-(2-((2S,4S)-1-(((methoxycarbonyl)amino)(tetr-
ahydro-2H-pyran-4-yl)acetyl)-4-methyl-2-pyrrolidinyl)-1H-imidazol-4-yl)-2--
naphthyl)phenyl)-1H-imidazol-2-yl)-4-methyl-1-pyrrolidinyl)-2-oxo-1-(tetra-
hydro-2H-pyran-4-yl)ethyl)carbamate
[1065] Example 146 (22 mg, 13%) was obtained and isolated from the
same reaction that produced Example 129 and as a diastereomer
thereof. LC-MS retention time2.86 min; calcd. for
C.sub.50H.sub.61N.sub.8O.sub.8: 901.46 Found m/z 901.42 [M+H]. LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped a PHENOMENEX.RTM. C18 2.0.times.50 mm column using a
SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The
elution conditions employed a flow rate of 0.8 mL/min, a gradient
of 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B, a
gradient time of 4 min, a hold time of 1 min and an analysis time
of 5 min where Solvent A was 5% MeOH/95% water/0.1% TFA and Solvent
B was 95% MeOH/5% water/0.1% TFA. MS data was determined using a
MICROMASS.RTM. Platform for LC in electrospray mode. .sup.1H NMR
(TFA salt, 500 MHz, MeOD) .delta. ppm 8.34/8.29 (s, 1H), 8.27 (s,
1H), 8.13 (d, J=7.63 Hz, 1H), 8.08 (d, J=8.6 Hz, 1H), 7.99-7.95 (m,
4H), 7.92-7.87 (m, 4H), 5.29-5.21 (m, 2H), 4.38 (t, J=7.3 Hz, 1H),
4.31-4.28 (m, 2H), 4.16-4.13 (m, 1H), 3.99-3.91 (m, 4H), 3.68 (s,
3H), 3.65/3.61 (s, 3H), 3.56-3.36 (m, 6H), 2.76-2.62 (m, 2H), 2.56
(br. s, 2H), 2.00-1.83 (m, 4H), 1.71 (d, J=12.8 Hz, 1H), 1.31-131.
(m, 7H), 1.26-1.2 (m, 6H).
##STR00746##
Example 147
Methyl
((1S)-2-((5S)-5-(4-(4-(6-(2-((3S)-2-((2S)-2-((methoxycarbonyl)amino-
)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-1-methyl-3-pyrazolidinyl)-1H-imidazo-
l-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-methyl-1-pyrazolidinyl)-2-o-
xo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
[1066] (S)-tert-Butyl
5-(4-(4-(6-(2-((S)-2-(tert-butoxycarbonyl)-1-methylpyrazolidin-3-yl)-1H-i-
midazol-4-yl)naphthalen-2-yl)phenyl)-1H-imidazol-2-yl)-2-methylpyrazolidin-
e-1-carboxylate (48.4 mg, 0.069 mmol) was dissolved into dioxane
(1.2 mL) and then 4M HCl (0.687 mL, 2.75 mmol) in dioxane was added
and the resulting slurry was stirred for 4 h. The reaction was
concentrated and the residue and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic
acid (37.3 mg, 0.172 mmol) were dissolved into DMF (1.0 mL) and
DIPEA (0.072 mL, 0.41 mmol). The reaction mixture was treated with
HATU (65 mg, 0.17 mmol) and the reaction was stirred at rt for 1 h.
The reaction was concentrated to dryness with a stream of nitrogen,
dissolved into MeOH, filtered and purified by prep HPLC (MeOH/water
with TFA buffer) to yield methyl
((1S)-2-((5S)-5-(4-(4-(6-(2-((3S)-2-((2S)-2-((methoxycarbonyl)amin-
o)-2-(tetrahydro-2H-pyran-4-yl)acetyl)-1-methyl-3-pyrazolidinyl)-1H-imidaz-
ol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2-methyl-1-pyrazolidinyl)-2--
oxo-1-(tetrahydro-2H-pyran-4-yl)ethyl)carbamate (9.2 mg) as a
yellow solid. LC-MS retention time 3.261 min; m/z 903.36 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph
equipped with a PHENOMENEX.RTM. Luna 3u C18 2.0.times.50 mm column
using a SPD-10AV UV-Vis detector at a detector wave length of 220
nM. The elution conditions employed a flow rate of 0.8 mL/min, a
gradient of 100% Solvent A/0% Solvent B to 0% Solvent A/100%
Solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time of 5 min where Solvent A was 10% MeOH/90%
H.sub.2O/0.1% trifluoroacetic acid and Solvent B was 10%
H.sub.2O/90% MeOH/0.1% trifluoroacetic acid. MS data was determined
using a MICROMASS.RTM. Platform for LC in electrospray mode.
.sup.1H NMR (400 MHz, MeOD) .delta. ppm 8.32 (s, 1H), 8.27 (s, 1H),
8.13 (d, J=8.5 Hz, 1H), 8.09 (d, J=8.8 Hz, 1H), 7.94-8.00 (m, 4H),
7.85-7.93 (m, 4H), 5.47 (q, J=8.1 Hz, 2H), 3.93-4.01 (m, 4H), 3.66
(s, 6H), 3.34-3.49 (m, 6H), 3.20-3.34 (m, 4H), 2.77 (s, 3H), 2.77
(s, 3H), 2.65-2.96 (m, 4H), 2.08 (br. s., 2H), 1.38-1.73 (m,
8H).
##STR00747##
Example 148
Methyl
((1R)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2R)-2-((methoxycarbonyl)ami-
no)-3-methylbutanoyl)amino)-2,2-dimethylpropyl)-1H-imidazol-4-yl)-2-naphth-
yl)phenyl)-1H-imidazol-2-yl)-2,2-dimethylpropyl)carbamoyl)-2-methylpropyl)-
carbamate
[1067] To a mixture of an HCl salt of
(S)-1-(5-(4-(6-(2-((S)-1-amino-2,2-dimethylpropyl)-1H-imidazol-5-yl)napht-
halen-2-yl)phenyl)-1H-imidazol-2-yl)-2,2-dimethylpropan-1-amine
(0.04 g, 0.061 mmol), (R)-2-(methoxycarbonylamino)-3-methylbutanoic
acid (0.021 g, 0.123 mmol) and HATU (0.051 g, 0.135 mmol) in DMF
was added DIPEA (0.064 mL, 0.368 mmol). The reaction mixture was
stirred at rt for 1 h. The reaction mixture was purified by reverse
phase HPLC to yield a TFA salt of methyl
((1R)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2R)-2-((methoxycarbonyl-
)amino)-3-methylbutanoyl)amino)-2,2-dimethylpropyl)-1H-imidazol-4-yl)-2-na-
phthyl)phenyl)-1H-imidazol-2-yl)-2,2-dimethylpropyl)carbamoyl)-2-methylpro-
pyl)carbamate (0.020 g) as a white solid. LC-MS (Cond. 1; shown
below): [M+H].sup.+821.55, Rt=3.316 min; .sup.1H NMR (400 MHz,
MeOD) ppm 8.34 (1H, s), 8.27 (1H, d, J=1.25 Hz), 8.11 (2H, dd,
J=15.94, 8.91 Hz), 7.84-8.02 (8H, m), 4.94 (2H, s), 4.07 (2H, dd,
J=7.78, 3.76 Hz), 3.60 (6H, d, J=9.29 Hz), 1.94-2.14 (2H, m), 1.16
(18H, d, J=5.27 Hz), 0.95-1.07 (12H, m).
[1068] Cond. 1: Column .dbd.PHENOMENEX.RTM., 2.0.times.50 mm, 3
.mu.m; Start % B=0; Final % B=100; Gradient time=4 min; Stop time=5
min; Flow Rate=0.8 mL/min; Wavelength=220 nm; Solvent A=0.1% TFA in
10% methanol/90% water; Solvent B=0.1% TFA in 90% methanol/10%
water; Oven temp.=40.degree. C.)
Example 149 and Example 150
[1069] Example 149 (a TFA salt of methyl
((1S)-1-(((1S)-1-(4-(4-(6-(2-((1S)-1-(((2S)-2-((methoxycarbonyl)amino)-3--
methylbutanoyl)amino)-2,2-dimethylpropyl)-1H-imidazol-4-yl)-2-naphthyl)phe-
nyl)-1H-imidazol-2-yl)-2,2-dimethylpropyl)carbamoyl)-2-methylpropyl)carbam-
ate) and Example 150 (a TFA salt of
N-((1S)-1-(4-(4-(6-(2-(1S)-2,2-dimethyl-1-((3-methylbutanoyl)amino)propyl-
)-1H-imidazol-4-yl)-2-naphthyl)phenyl)-1H-imidazol-2-yl)-2,2-dimethylpropy-
l)-3-methylbutanamide) were prepared from
2-bromo-1-(4-(6-(2-bromoacetyl)naphthalen-2-yl)phenyl)ethanone and
the appropriate starting materials, obtained from commercial
sources, by employing the procedures described for the synthesis of
Example 148. Cond. 2: Column=Sunfire, C18, 3.0.times.150 mm, 3.5
.mu.m; Start % B=0; Final % B=100; Gradient time=15 min; Stop
time=18 min; Flow Rate=1 mL/min; Wavelength 1=220 nm; Wavelength
2=254 nm; Solvent A=0.1% TFA in 5% MeCN/95% water; Solvent B=0.1%
TFA in 95% MeCN/5% water
TABLE-US-00011 ##STR00748## Analytical Example R conditions 149
##STR00749## LC (Cond. 2): >95% homogeneity index. LC-MS (Cond.
1): [M + H].sup.+ 821.55, R.sub.t = 3.348 min. 150 ##STR00750## LC
(Cond. 2): >95% homogeneity index. LC-MS (Cond. 1): [M +
H].sup.+ 675.51, R.sub.t = 3.488 min.
Example 154
##STR00751##
[1071] In a 500-mL reactor Example 151 (commercially available,
17.5 g, 1.00 equiv) was dissolved in THF (87.5 mL). The resulting
solution was cooled to -75.degree. C. and 1.5M DIBAL-H in toluene
(61.3 mL, 1.5 equiv) was charged while maintaining the temperature
below -70.degree. C. The resulting solution was stirred at
-70.degree. C. for 1 hour. Trifluoroacetic acid (2.3 mL, 0.5 equiv)
was charged over 10 minutes maintaining the internal temperature
below -70.degree. C. Triethylamine (51.3 mL, 6 equiv) was then
charged over 15 minutes maintaining the internal temperature below
-70.degree. C. Trifluoroacetic anhydride (11.2 mL, 1.3 equiv) was
charged over 10 minutes maintaining the internal temperature below
-70.degree. C. The reaction was then allowed to warm to room
temperature over 90 minutes and quenched via inverse addition to a
solution of 20 wt % aqueous citric acid monohydrate (96.6 g, 1.5
equiv) while maintaining a temperature below 15.degree. C. The
resulting mixture was stirred at room temperature for 2 hours then
the lower aqueous layer was discarded. The product rich organic
layer was washed twice with 70 mL saturated aqueous sodium
bicarbonate. Solid sodium bicarbonate (1.7 g, 0.1 g/g Example 146)
was charged and the solution was solvent exchanged into pure
toluene under vacuum to provide Example 152 as a solution in 2 L/kg
toluene.
[1072] A solution of Example 152 (16.5 g theoretical from Example
151) in 33 mL toluene was polish filtered into a 250 mL reactor.
Trifluorotoluene (50 mL) and chloroiodomethane (43.2 g, 4.0 equiv)
were then charged and the resulting solution cooled to -20.degree.
C. 1.1M Diethylzinc in toluene (111 mL, 2.0 equiv) was charged
while maintaining the internal temperature<-8.degree. C. The
resulting solution was stirred at -15 to -20.degree. C. for 14
hours. The reaction mixture was warmed to 0.degree. C. then
quenched via inverse addition to a solution of 20 wt % aqueous
citric acid (135.7 g, 2.3 equiv). The reactor was rinsed with
toluene (82 mL) and the rinse added to the quench solution. The
resulting biphasic mixture was stirred for 20 minutes then the
lower aqueous layer was split and discarded. The rich organic was
washed twice with 60 mL 13 wt % aqueous NaCl followed by 60 mL
saturated NaHCO.sub.3. The resulting solution was solvent exchanged
into pure IPA under vacuum to provide Example 153 as a solution in
10 L/kg IPA.
[1073] A 250 mL reactor was charged with a solution of Example 153
(147 mL, 14.7 g theoretical from Example 151) in IPA. The solution
was warmed to 35.degree. C. and solid sodium hydroxide (6.2 g, 3.0
equiv) was added. The resulting mixture was stirred at 35.degree.
C. overnight. Water (44 mL) was added and the organic solvents
removed under vacuum. MTBE (145 ml) was added and the pH adjusted
to 3.0 with 6N aqueous HCl. The aqueous layer was split and
discarded. The product rich organic was washed with 60 mL water
then azeotropically dried under vacuum via constant volume addition
of MTBE. The solution was concentrated to 55 mL and stirred at
50.degree. C. for 30 minutes. The solution was cooled to room
temperature over 1 hour during which time a slurry formed. Heptane
(90 mL) was charged over 90 min and the resulting slurry aged for 1
h. The solids were collected on a medium glass frit and washed with
22.5 mL 3:1 heptane:MTBE followed by 22.5 mL heptane. The tan solid
was dried in a 50.degree. C. vacuum oven to provide 5.48 g (46%)
Example 154 with 94.9 LCAP purity.
[1074] The crude Example 154 was dissolved in 55 mL MTBE at
50.degree. C. The resulting solution was concentrated to 20 mL and
cooled to room temperature over 1 hour. Heptane (33 mL) was then
added over 90 minutes. The resulting solids were collected on a
medium glass frit, washed with heptane (15 mL), and dried in a
50.degree. C. vacuum oven to provide 4.45 g (98.8 AP, 98.8% chiral
purity, 37% from Example 151) of the desired product as a tan
powder.
[1075] It is expected that the above reaction sequence (for the
formation of Example 154) would be general enough to tolerate other
functionality as well. For example, the tert-butoxycarbonyl group
on the C2 carbon of Example 147 could potentially be replaced with
an amide or another ester. Likewise, the tert-butoxycarbonyl
protecting group on the nitrogen could be replaced by other
protecting groups (such as other carbamates, amides, alkyl, aryl)
or could be absent entirely (replaced with a hydrogen).
Biological Activity
[1076] An HCV Replicon assay was utilized in the present
disclosure, and was prepared, conducted and validated as described
in commonly owned PCT/US2006/022197 and in O'Boyle et al.,
Antimicrob. Agents Chemother., 49(4):1346-1353 (April 2005). Assay
methods incorporating luciferase reporters have also been used as
described (Apath.com).
[1077] HCV-neo replicon cells and replicon cells containing
resistance substitutions in the NS5A region were used to test the
currently described family of compounds. The compounds were
determined to have differing degrees of reduced inhibitory activity
on cells containing mutations vs. the corresponding inhibitory
potency against wild-type cells. Thus, the compounds of the present
disclosure can be effective in inhibiting the function of the HCV
NS5A protein and are understood to be as effective in combinations
as previously described in application PCT/US2006/022197 and
commonly owned WO 04/014852. It should be understood that the
compounds of the present disclosure can inhibit multiple genotypes
of HCV. Table 2 shows the EC.sub.50 (Effective 50% inhibitory
concentration) values of representative compounds of the present
disclosure against the HCV 1b genotype. In one embodiment,
compounds of the present disclosure are inhibitory versus 1a, 1b,
2a, 2b, 3a, 4a, and 5a genotypes. EC.sub.50 values against HCV 1b
are as follows: A=0.4 pM-10 pM; B=10.1 pM-1.00 nM; and C=1.01 nM-1
.mu.M.
[1078] The compounds of the present disclosure may inhibit HCV by
mechanisms in addition to or other than NS5A inhibition. In one
embodiment the compounds of the present disclosure inhibit HCV
replicon and in another embodiment the compounds of the present
disclosure inhibit NS5A.
TABLE-US-00012 TABLE 2 Example Number (unless noted otherwise) EC50
(.mu.M) Range Intermediate 52 C Intermediate 70 C Intermediate 114
C 1 C 2 2.18E-06 A 3 B 4 C 5 B 6 A 7 B 8 B 9 A 10 A 11 1.82E-05 B
12 C 13 C 14 C 15 B 16 A 17 C 18 A 19 B 20 A 21 A 22 B 23 8.88E-04
B 24 B .sup. 25A C .sup. 25B C 26 B 27 4.77E-06 A 28 B 29 A 30 A 31
B 32 B 33 A 34 A 35 A 36 A 37 A 38 4.21E-05 B 39 B 40 A 41 A 42 B
43 A 44 A 45 B 46 0.17 C 47 A 48 B 49 A 50 B 51 B 52 5.90E-06 A 53
1.81E-03 C 54 A 55 A 56 A 57 A 58 8.49E-04 B 59 A 60 A 61 A 62 A 63
A 64 A 65 9.97E-07 A 66 A 67 A 68 A 69 A 70 A 71 2.11E-06 A 72 A 73
A 74 75 1.23E-04 C 76 77 A 78 A 79 A 80 1.08E-06 A 81 A 82 4.22E-05
B 83 A 84 A 85 A 86 A 87 A 88 A 89 A 90 A 91 A 92 A 93 0.04 C 94 A
95 A 96 B 97 A 98 B 99 A 100 A 101 A 102 A 103 A 104 2.01E-06 A 105
A 106 A 107 A 108 109 0.18 C 110 A 111 A 112 A 113 114 B 115 A 116
B 117 118 3.74E-05 B 119 A 120 A 121 A 122 A 123 A 124 B 125
1.36E-04 B 126 6.54E-04 B 127 A 128 A 129 B 130 A 131 -- 132 A 133
A 134 A 135 B 136 A 137 A 138 A 139 A 140 A 141 A 142 A 143
1.05E-06 A 144 1.77E-05 B 145 A 146 A 147 A 148 8.43E-03 C 149 B
150 C
[1079] It will be evident to one skilled in the art that the
present disclosure is not limited to the foregoing illustrative
examples, and that it can be embodied in other specific forms
without departing from the essential attributes thereof. It is
therefore desired that the examples be considered in all respects
as illustrative and not restrictive, reference being made to the
appended claims, rather than to the foregoing examples, and all
changes which come within the meaning and range of equivalency of
the claims are therefore intended to be embraced therein.
* * * * *