U.S. patent application number 10/621979 was filed with the patent office on 2004-05-13 for amide compounds and pharmaceutical compositions for inhibiting protein kinases, and methods for their use.
Invention is credited to Bender, Steven Lee, Bhumaralkar, Dilip, Collins, Michael Raymond, Cripps, Stephen James, Deal, Judith Gail, Jia, Lei, Nambu, Mitchell David, Palmer, Cynthia Louise, Peng, Zhengwei, Varney, Michael David.
Application Number | 20040092747 10/621979 |
Document ID | / |
Family ID | 22647011 |
Filed Date | 2004-05-13 |
United States Patent
Application |
20040092747 |
Kind Code |
A1 |
Bender, Steven Lee ; et
al. |
May 13, 2004 |
Amide compounds and pharmaceutical compositions for inhibiting
protein kinases, and methods for their use
Abstract
Amide compounds that modulate and/or inhibit the activity of
certain protein kinases are described. These compounds and
pharmaceutical compositions containing them are capable of
mediating tyrosine kinase signal transduction in order to modulate
and/or inhibit unwanted cell proliferation. The invention is also
directed to the therapeutic or prophylactic use of pharmaceutical
compositions containing such compounds, and to methods of treating
cancer as well as other disease states associated with unwanted
angiogenesis and/or cellular proliferation, such as diabetic
retinopathy, neovascular glaucoma, rheumatoid arthritis, and
psoriasis, by administering effective amounts of such
compounds.
Inventors: |
Bender, Steven Lee;
(Oceanside, CA) ; Bhumaralkar, Dilip; (San Diego,
CA) ; Collins, Michael Raymond; (San Diego, CA)
; Cripps, Stephen James; (San Diego, CA) ; Deal,
Judith Gail; (Wildomar, CA) ; Jia, Lei; (San
Diego, CA) ; Nambu, Mitchell David; (San Diego,
CA) ; Palmer, Cynthia Louise; (La Mesa, CA) ;
Peng, Zhengwei; (San Diego, CA) ; Varney, Michael
David; (Solana Beach, CA) |
Correspondence
Address: |
AGOURON PHARMACEUTICALS, INC.
10350 NORTH TORREY PINES ROAD
LA JOLLA
CA
92037
US
|
Family ID: |
22647011 |
Appl. No.: |
10/621979 |
Filed: |
July 17, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10621979 |
Jul 17, 2003 |
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09764306 |
Jan 19, 2001 |
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6635641 |
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60177059 |
Jan 21, 2000 |
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Current U.S.
Class: |
548/131 ;
548/136; 548/262.2 |
Current CPC
Class: |
C07D 241/12 20130101;
C07D 213/56 20130101; C07D 231/38 20130101; C07D 217/14 20130101;
C07D 217/16 20130101; C07D 401/12 20130101; A61P 35/00 20180101;
C07D 249/14 20130101; C07D 217/22 20130101; C07D 241/18 20130101;
C07D 233/56 20130101; C07D 471/04 20130101; C07D 401/06 20130101;
A61P 43/00 20180101; C07D 231/12 20130101; C07D 487/04 20130101;
C07D 249/08 20130101; C07D 405/12 20130101; C07D 213/30 20130101;
C07D 405/04 20130101; C07D 237/28 20130101; C07D 401/14
20130101 |
Class at
Publication: |
548/131 ;
548/136; 548/262.2 |
International
Class: |
C07D 271/12; C07D
285/14; C07D 249/08 |
Claims
What is claimed:
1. A compound represented by the Formula I: 352wherein: R.sup.1 is
a moiety represented by the formula 353 where Z is selected from
the group consisting of CH and NH, and Q is a moiety such that
R.sup.1 is a substituted or unsubstituted monocyclic or bicyclic
heteroaryl which has at least two carbon atoms in the heteroaryl
ring system; X is selected from the group consisting of CH.sub.2,
O, S, and NH; Y is selected from the group consisting of CH.sub.2,
O, and S, provided that at least one of X and Y is CH.sub.2, or X
and Y together with the bond there-between form a cyclopropyl;
R.sup.2 and R.sup.3 are independently selected from the group
consisting of hydrogen, methyl, halogen, trifluoromethyl, and
cyano; and R.sup.4 is selected from the group consisting of 354
where R.sup.5 is selected from the group consisting of substituted
and unsubstituted aryl, heteroaryl, cycloalkyl, heterocycloalkyl,
O--R.sup.7, NR.sup.8R.sup.9, C.sub.1-C.sub.8 alkyl, and monocyclic
heterocycloalkyl, R.sup.6 is selected from the group consisting of
substituted and unsubstituted aryl, heteroaryl, cycloalkyl,
heterocycloalkyl, alkenyl, O--R.sup.7, C(O)R.sup.7,
NR.sup.8R.sup.9, C.sub.2-C.sub.8 alkyl, and monocyclic
heterocycloalkyl, where R.sup.7 is selected from the group
consisting of substituted and unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, R.sup.8 is selected from
the group consisting of hydrogen, and substituted and unsubstituted
alkyl, and R.sup.9 is selected from the group consisting of
substituted and unsubstituted alkyl, aryl, heteroaryl, cycloalkyl,
and heterocycloalkyl; or a pharmaceutically acceptable prodrug,
pharmaceutically active metabolite, or pharmaceutically acceptable
salt thereof.
2. A compound according to claim 1, wherein R.sup.1 is a
substituted or unsubstituted heteroaryl group selected from the
group consisting of: 355X is selected from the group consisting of
CH.sub.2, O, and S; Y is selected from the group consisting of
CH.sub.2 and S, provided that at least one of X and Y is CH.sub.2;
R.sup.2 and R.sup.3 are independently selected from the group
consisting of hydrogen, methyl, fluorine, and chlorine, and R.sup.4
is selected from the group consisting of 356 where R.sup.5 is
selected from the group consisting of substituted and unsubstituted
aryl, heteroaryl, cycloalkyl, heterocycloalkyl, O--R.sup.7,
NR.sup.8R.sup.9, C.sub.1-C.sub.8 alkyl and monocyclic
heterocycloalkyl, R.sup.6 is selected from the group consisting of
substituted and unsubstituted aryl, heteroaryl, cycloalkyl,
heterocycloalkyl, alkenyl, O--R.sup.7, C(O)R.sup.7,
NR.sup.8R.sup.9, C.sub.2-C.sub.8 alkyl, and monocyclic
heterocycloalkyl, where R.sup.7 is selected from the group
consisting of substituted and unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, R.sup.8 is selected from
the group consisting of hydrogen and substituted and unsubstituted
alkyl, and R.sup.9 is selected from the group consisting of
substituted and unsubstituted alkyl, aryl, heteroaryl, cycloalkyl,
and heterocycloalkyl; or a pharmaceutically acceptable prodrug,
pharmaceutically active metabolite, or pharmaceutically acceptable
salt thereof.
3. A compound represented by the Formula II: 357wherein: X is
selected from the group consisting of CH.sub.2, O, and S; Y is is
selected from the group consisting of CH.sub.2 and S, provided that
at least one of X and Y is CH.sub.2; R.sup.2 and R.sup.3 are
independently selected from the group consisting of hydrogen,
methyl, fluorine, and chlorine; R.sup.4 is selected from the group
consisting of 358 where R.sup.5 and R.sup.6 are each independently
selected from the group consisting of substituted and unsubstituted
aryl and heteroaryl; and R.sup.10 is selected from the group
consisting of substituted and unsubstituted alkenyl, aryl,
heteroaryl, and HNR.sup.9, where R.sup.9 is selected from the group
consisting of substituted and unsubstituted alkyl, aryl,
heteroaryl, cycloalkyl, and heterocycloalkyl; or a pharmaceutically
acceptable prodrug, pharmaceutically active metabolite, or
pharmaceutically acceptable salt thereof.
4. A compound according to claim 3, wherein R.sup.5 and R.sup.6 are
each independently selected from the group consisting of
substituted and unsubstituted aryl; or a pharmaceutically
acceptable salt, a pharmaceutically acceptable prodrug, or a
pharmaceutically active metabolite thereof, or a pharmaceutically
acceptable salt of said metabolite.
5. A compound according to claim 3, wherein R.sup.5 and R.sup.6 are
each independently selected from the group consisting of
substituted and unsubstituted heteroaryl; or a pharmaceutically
acceptable salt, a pharmaceutically acceptable prodrug, or a
pharmaceutically active metabolite thereof, or a pharmaceutically
acceptable salt of said metabolite.
6. A compound represented by the Formula III: 359wherein: X is
selected from the group consisting of CH.sub.2, O, S, and NH; Y is
selected from the group consisting of CH.sub.2, O, and S, provided
that at least one of X and Y is CH.sub.2, or X and Y together with
the bond there-between form a cyclopropyl; R.sup.2 and R.sup.3 are
independently selected from the group consisting of hydrogen,
methyl, halogen, trifluoromethyl, and cyano; and R.sup.4 is
selected from the group consisting of 360 where R.sup.5 is selected
from the group consisting of substituted and unsubstituted aryl,
heteroaryl, cycloalkyl, heterocycloalkyl, O--R.sup.7,
NR.sup.8R.sup.9, C.sub.1-C.sub.8 alkyl, and monocyclic
heterocycloalkyl, R.sup.6 is selected from the group consisting of
substituted and unsubstituted aryl, heteroaryl, cycloalkyl,
heterocycloalkyl, alkenyl, O--R.sup.7, C(O)R.sup.7,
NR.sup.8R.sup.9, C.sub.2-C.sub.8 alkyl, and monocyclic
heterocycloalkyl, where R.sup.7 is selected from the group
consisting of substituted and unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, R.sup.8 is selected from
the group consisting of hydrogen and substituted and unsubstituted
alkyl, and R.sup.9 is selected from the group consisting of
substituted and unsubstituted alkyl, aryl, heteroaryl, cycloalkyl,
and heterocycloalkyl; or a pharmaceutically acceptable prodrug,
pharmaceutically active metabolite, or pharmaceutically acceptable
salt thereof.
7. A compound according to claim 6, wherein: X is selected from the
group consisting of CH.sub.2, O, and S; Y is selected from the
group consisting of CH.sub.2 and S, provided that at least one of X
and Y is CH.sub.2; R.sup.2 and R.sup.3 are independently selected
from the group consisting of hydrogen, methyl, fluorine, and
chlorine; and R.sup.4 is selected from the group consisting of 361
where R.sup.5 and R.sup.6 are each independently selected from the
group consisting of substituted and unsubstituted aryl and
heteroaryl; or a pharmaceutically acceptable salt, a
pharmaceutically acceptable prodrug, or a pharmaceutically active
metabolite thereof, or a pharmaceutically acceptable salt of said
metabolite.
8. A compound according to claim 7, wherein R.sup.5 and R.sup.6 are
each independently selected from the group consisting of
substituted and unsubstituted aryl; or a pharmaceutically
acceptable salt, a pharmaceutically acceptable prodrug, or a
pharmaceutically active metabolite thereof, or a pharmaceutically
acceptable salt of said metabolite.
9. A compound according to claim 7, wherein R.sup.5 and R.sup.6 are
each independently selected from the group consisting of
substituted and unsubstituted heteroaryl; or a pharmaceutically
acceptable salt, a pharmaceutically acceptable prodrug, or a
pharmaceutically active metabolite thereof, or a pharmaceutically
acceptable salt of said metabolite.
10. A compound according to claim 7, wherein: X is CH.sub.2; Y is
S; R.sup.2 and R.sup.3 are independently selected from the group
consisting of hydrogen, methyl, fluorine, and chlorine; and R.sup.4
is selected from the group consisting of 362 where R.sup.5 and
R.sup.6 are each independently selected from the group consisting
of substituted and unsubstituted aryl and heteroaryl; or a
pharmaceutically acceptable salt, a pharmaceutically acceptable
prodrug, or a pharmaceutically active metabolite thereof, or a
pharmaceutically acceptable salt of said metabolite.
11. A compound according to claim 10, wherein R.sup.5 and R.sup.6
are each independently selected from the group consisting of
substituted and unsubstituted aryl; or a pharmaceutically
acceptable salt, a pharmaceutically acceptable prodrug, or a
pharmaceutically active metabolite thereof, or a pharmaceutically
acceptable salt of said metabolite.
12. A compound selected from the group consisting of
N-(3,4,5-Trimethoxyphenyl)-3-[(pyrazin-2-yl)sulfanylmethyl]benzamide;
N-(3,4,5-Trimethoxyphenyl)-3-[(5-amino-2H-[1,2,4]triazol-3-yl)sulfanylmet-
hyl]benzamide;
N-(4-Isopropyl-3-methylphenyl)-3-[(pyrazin-2-yl)sulfanylmet-
hyl]benzamide;
N-(4-Isopropyl-3-methylphenyl)-3-[(5-amino-2H-[1,2,4]triazo-
l-3-yl)sulfanylmethyl]benzamide;
N-(4-Isopropyl-3-methylphenyl)-3-[(1H-pyr-
azolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]benzamide;
N-(2-Methylquinolin-6-yl)-3-[(pyrazin-2-yl)sulfanylmethyl]benzamide;
N-(3-Isopropylphenyl)-3-[(pyrazin-2-yl)sulfanylmethyl]benzamide;
N-(3,5-Dibromo-4-methylphenyl)-3-[(pyrazin-2-yl)sulfanylmethyl]benzamide;
N-(3,4,5-Trimethoxyphenyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylm-
ethyl]benzamide;
N-(3,4,5-Trimethoxyphenyl)-3-[(1H-pyrazolo[3,4-d]pyrimidi-
n-4-yl)sulfanylmethyl]benzamide;
N-(Quinolin-6-yl)-3-[(1H-pyrazolo[3,4-d]p-
yrimidin-4-yl)sulfanyl-methyl]benzamide;
N-(5-Methylisoxazol-3-yl)-3-[(1H--
pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]benzamide;
N-(Pyridin-4-yl)methyl-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethy-
l]benzamide;
N-(1,3-Benzodioxyl-5-ylmethyl)-3-[(1H-pyrazolo[3,4-d]pyrimidi-
n-4-yl)-sulfanylmethyl]benzamide;
N-(2-Methoxybenzyl)-3-[(1H-pyrazolo[3,4--
d]pyrimidin-4-yl)-sulfanylmethyl]benzamide;
N-(2-Phenylethyl)-3-[(1H-pyraz-
olo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]benzamide;
N-(2-Methoxyphenyl)-3-[-
(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-sulfanylmethyl]benzamide;
N-[3-(N-Methyl-N-phenylamino)propyl]-3-[(5-methyl-1H-1,2,4-triazol-3-yl)
sulfanylmethyl]benzamide;
N-(1,3-Benzodioxyl-5-ylmethyl)-3-[(5-methyl-1H--
1,2,4-triazol-3-yl)sulfanylmethyl]benzamide;
N-[4-cyano-3-(trifluoromethyl-
)phenyl]-3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl-sulfanyl)
methyl]benzamide;
N-(3,3-Diphenylpropyl)-3-{[(5-methyl-1H-1,2,4-triazol-3-yl)-sulfanyl]meth-
yl}benzamide;
3-[(5-Methyl-1H-1,2,4-triazol-3-yl)-sulfonyl]methyl}-N-phene-
thylbenzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-is-
opropylphenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmet-
hyl]-N(3-trifluoromethyl-5-methoxyphenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3,5-bis-trifluorom-
ethylphenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethy-
l]-N(3-t-butylphenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulf-
anylmethyl]-N(4-isopropylphenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidi-
n-4-yl)sulfanylmethyl]-N(4-trifluoromethoxyphenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3,5-dimethylphenyl-
)-benzamide:
3[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-(2-hy-
droxyethyl)phenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfany-
lmethyl]-N(4-dimethylaminophenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimid-
in-4-yl)sulfanylmethyl]-N(3-trifluoromethylsulfonyl
phenyl)-benzamide;
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-dimethylaminophe-
nyl)-benzamide;
3-[(5-Cyanoamino-2H-[1,2,4]triazol-3-yl)sulfanylmethyl]-N--
(3,4,5-trimethoxyphenyl) benzamide;
3-[(5-(Methoxycarbonylamino)-2H-[1,2,4-
]triazol-3-yl)sulfanylmethyl]-N-(3,4,5-trimethoxyphenyl)benzamide;
N-(3,4,5-Trimethoxyphenyl)-3-[(5-acetylamino-2H-[1,2,4]triazol-3-yl)sulfa-
nylmethyl]benzamide;
N-(4-Isopropyl-3-methylphenyl)-3-[(pyrazin-2-yl)methy-
lsulfanyl]benzamide;
N-(2-Methylquinolin-6-yl)-3-[(pyrazin-2-yl)methylsulf-
anyl]benzamide;
N-(2-Methyl-quinolin-6-yl)-3-(pyridin-3-ylmethylsulfanyl)--
benzamide dihydrochloride;
N-(2-methyl-quinolin-6-yl)-3-[{5-(phenylamino)--
2-H-pyrazol-3-yl}methylsulfanyl]benzamide;
N-(3,4,5-trimethoxyphenyl)-3-[2-
-(5-phenylamino-2H-pyrazol-3-yl)ethyl]benzamide;
3-[{5-((E)-2-(4-Hydroxy-3-
-methoxyphenyl)ethenyl)-2H-pyrazol-3-yl}-methylsulfanyl]-N-(2-methylquinol-
in-6-yl)benzamide;
3-[5-(2-(3,4-Dimethoxyphenyl)ethenyl)-2H-pyrazol-3-yl)m-
ethylsulfanyl]-N-(2-methylquinolin-6-yl)benzamide;
3-(2-{5-[(E)-2-(3,4-Dim-
ethoxyphenyl)ethenyl]-2H-pyrazol-3-yl}-ethyl)-N-(3-methyl-4-isopropylpheny-
l)-benzamide;
4-Fluoro-3-[{5-((E)-1-propenyl)-2H-pyrazol-3-yl}methoxy]-N-[-
4-(pyrrolidin-1-yl)-3-trifluoromethylphenyl]benzamide;
3-(2-{5-[(E)-2-(3,4-Dimethoxyphenyl)ethenyl]-2H-pyrazol-3-yl}-ethyl)-N-(3-
-methyl-4-isopropylphenyl)-benzamide;
N-(4-Isopropyl-3-methyl-phenyl)-3-{2-
-[5-(4-(methylsulfamoyl)-phenylamino)-2H-pyrazol-3-yl]-ethyl}-benzamide;
N-(2-Methylquinolin-6-yl)-3-[2-(5-phenylamino-2H-pyrazol-3-yl)ethyl]benza-
mide;
N-(4-isopropyl-3-methylphenyl)-3-[2-(5-phenylamino-2H-pyrazol-3-yl)e-
thyl]benzamide;
N-(4-Isopropyl-3-methyl-phenyl)-3-{2-[5-(6-methoxypyridin--
3-yl)amino-2H-pyrazol-3-yl]-ethyl}-benzamide;
N-(4-Dimethylamino-3-trifluo-
romethylphenyl)-3-{2-[5-(6-methoxypyridin-3-yl)amino-2H-pyrazol-3-yl]ethyl-
}-benzamide;
N-(6-Dimethylamino-5-trifluoromethylpyridin-3-yl)-3-{2-[5-(6--
methoxypyridin-3-yl)amino-2H-pyrazol-3-yl]ethyl}-benzamide;
N-(3,5-Dichloro-4-dimethylaminophenyl)-3-{2-[5-(6-methoxy-pyridin-3-yl)am-
ino-2H-pyrazol-3-yl]ethyl}benzamide;
3-{2-[5-(6-Methoxypyridin-3-yl)amino--
2H-pyrazol-3-yl]-ethyl}-N-(4-pyrrolidin-1-yl-3-trifluoromethylphenyl)benza-
mide;
3-{2-[5-(6-Methoxypyridin-3-yl)amino-2H-pyrazol-3-yl]-ethyl}-N-[4-(4-
-t-butoxycarbonylpiperazin-1-yl)-3-trifluoromethylphenyl]benzamide;
3-{2-[5-(6-Methoxypyridin-3-yl)amino)-2H-pyrazol-3-yl]ethyl}-N-(4-piperaz-
in-1-yl-3-trifluoromethylphenyl)benzamide;
4-Fluoro-3-[{5-(pyridin-3-yl)am-
ino-2H-pyrazol-3-yl}methoxy]-N-[((4-pyrrolidin-1-yl)-3-trifluoromethylphen-
yl)benzamide;
N-(4-Isopropyl-3-methyl-phenyl)-3-[2-(5-phenylamino-2-H-pyra-
zol-3-yl)-cyclopropyl]-benzamide;
3-[({3-[(E)-2-(4-hydroxy-3-methoxyphenyl-
)ethenyl]-1H-pyrazol-5-yl}methyl)amino]-N-(3-methyl-4-isopropylphenyl)benz-
amide;
3-[({5-[(E)-2-(4-hydroxy-3-methoxyphenyl)ethenyl]-1H-pyrazol-3-yl}m-
ethyl)amino]-N-phenyl)benzamide;
4-Fluoro-N-[4-(imidazol-1-yl)-3-trifluoro-
methylphenyl]-3-[5-(6-methoxypyridin-3-yl)amino-2H-pyrazol-3-ylmethoxy]-be-
nzamide;
4-Fluoro-3-[5-(6-methoxy-pyridin-3-yl)amino-2H-pyrazol-3-yl]metho-
xy-N-(4-pyrrolidin-1-yl-3-trifluoromethyl-phenyl)-benzamide;
4-Fluoro-3-[5-(6-methoxypyridin-3-yl)amino-2H-pyrazol-3-yl]methoxy-N-(3-m-
ethoxy-5-trifluoromethyl-phenyl)-benzamide;
N-(4-Isopropyl-3-methyl-phenyl-
)-3-(Isoquinolin-4-yl)methoxy-benzamide;
3-(Isoquinolin-4-yl)methoxy-N-(3,- 4,5-trimethoxyphenyl)benzamide
hydrochloride; 3-(Isoquinolin-4-yl)methoxy--
N-(2-methyl-quinolin-6-yl)-benzamide hydrochloride;
3-(Isoquinolin-4-yl)methoxy-N-(2-methyl-4-methylsulfanyl-quinolin-6-yl)-b-
enzamide hydrochloride;
3-(Pyridin-3-yl)methoxy-N-(3,4,5-trimethoxyphenyl)- benzamide;
N-(Naphthalen-2-yl)-3-(pyridin-3-yl)methoxybenzamide;
N-(1-Allyl-1H-indol-5-yl)-3-(pyridin-3-yl)methoxy-benzamide;
3-(Pyridin-3-yl)methoxy-N-quinolin-6-yl-benzamide;
N-(2-Methyl-quinolin-6-yl)-3-(pyridin-3-yl)methoxy-benzamide;
N-(4-Isopropyl-3-methyl-phenyl)-4-fluoro-3-(Isoquinolin-4-yl)methoxy-benz-
amide;
N-(4-Isopropyl-3-methyl-phenyl)-4-methyl-3-(Isoquinolin-4-yl)methox-
y-benzamide;
N-(4-Isopropyl-3-methyl-phenyl)-4-chloro-3-(Isoquinolin-4-yl)-
methoxy-benzamide;
3-(6-Aminopyridin-3-yl)methoxy-N-(4-Isopropyl-3-methyl--
phenyl)benzamide;
3-(6-Aminopyridin-3-yl)methoxy-N-(2-methyl-quinolin-6-yl-
)-benzamide;
3-(6-Acetylaminopyridin-3-yl)methoxy-N-(2-methyl-quinolin-6-y-
l)-benzamide;
3-(6-Acetylaminopyridin-3-yl)methoxy-N-(4-isopropyl-3-methyl-
-phenyl)-benzamide;
4-Fluoro-N-(1,2,3,4-tetrahydroquinolin-6-yl)-3-(isoqui-
nolin-4-yl-methoxy)-benzamide bistrifluoroacetic acid salt;
N-(2,2-difluorobenzo[1,3]dioxol-4-yl-ethyl)-benzamide
trifluoroacetic acid salt;
4-Fluoro-N-(2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-3-(isoqu-
inolin-4-yl-methoxy)-benzamide bistrifluoroacetic acid salt;
N'-{4-[3-(4-Isopropyl-3-methyl-phenylcarbamoyl)-phenoxy;
N-(4-Isopropyl-3-methyl-phenyl)-3-{1-[N'-(3-methoxy-benzylidene)-hydrazin-
o]-isoquinolin-4-ylmethoxy}-benzamide;
N-(3,5-Diallyl-4-methyl-phenyl)-3-(-
isoquinolin-4-ylmethoxy)-benzamide;
N-(3,5-Dibromo-4-methyl-phenyl)-3-(iso-
quinolin-4-ylmethoxy)-benzamide;
3-(Isoquinolin-4-ylmethoxy)-N-(5,5,8,8-te-
tramethyl-5,6,7,8-tetrahydro-naphthalen-2-yl)-benzamide;
3-(Isoquinolin-4-ylmethoxy)-N-(3-trifluoromethoxy-phenyl)-benzamide;
N-(2,4-Dimethylquinolin-6-yl)-3-(isoquinolin-4-ylmethoxy)-benzamide;
3-(Isoquinolin-4-ylmethoxy)-benzoic acid
N'-(4-trifluoromethyl-phenyl)-hy- drazide;
N-Benzyloxy-3-(isoquinolin-4-ylmethoxy)-benzamide;
3-(Isoquinolin-4-ylmethoxy)-benzoic acid N'-phenyl-hydrazide;
N-(5,7-dimethyl
1,8]naphthydrin-2-yl)-3-(isoquinolin-4-ylmethoxy)-benzami- de;
3-(Isoquinolin-4-ylmethoxy)-N-(1,1,3,3-tetramethyl-1,3-dihydroisobenzo-
furan-5-yl)-benzamide;
N-(3,5-Dichloro-4-pyrrolidin-1-yl-phenyl)-4-fluoro--
3-(pyridin-3-ylmethoxy)-benzamide;
4-Fluoro-N-(4-morpholin-4-yl-3-trifluor-
omethyl-phenyl)-3-(pyridin-3-ylmethoxy)-benzamide;
4-Fluoro-N-[4-(piperazi-
n-1-yl)-3-trifluoromethylphenyl-3-3-(pyridin-3-yl)methoxybenzamide;
4-Fluoro-N-(4-morpholin-4-yl-3-trifluoromethyl-phenyl)-3-(isoquinolin-4-y-
lmethoxy)-benzamide;
4-Fluoro-N-(4-piperazin-1-yl-3-trifluoromethyl-phenyl-
)-3-(isoquinolin-4-ylmethoxy)-benzamide;
4-Fluoro-N-(4-morpholin-4-yl-3-tr-
ifluoromethyl-phenyl)-3-(quinolin-3-ylmethoxy)-benzamide;
4-Fluoro-N-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-3-(quinolin-3-ylme-
thoxy)-benzamide;
N-(3,5-Dichloro-4-morpholin-4-yl-phenyl)-4-fluoro-3-(pyr-
idin-3-ylmethoxy)-benzamide;
N-(3,5-Dichloro-4-piperazin-1-yl-phenyl)-4-fl-
uoro-3-(pyridin-3-ylmethoxy)-benzamide;
4-Fluoro-N-[4-(piperazin-1-yl)-3-t-
rifluoromethylphenyl]-3-(pyridin-3-yl)methoxybenzamide;
4-Fluoro-N-(4-(imidazol-1-yl-3-trifluoromethylphenyl]-3-(pyridin-3-yl)met-
hoxybenzamide;
4-Fluoro-N-(4-pyrazol-1-yl-3-trifluoromethyl-phenyl)-3-(pyr-
idin-3-ylmethoxy)-benzamide;
4-Fluoro-3-(pyridin-3-ylmethoxy)-N-(4-[1,2,4]-
triazol-1-yl-3-trifluoromethyl-phenyl)-benzamide;
N-(3,5-Dichloro-4-imidaz-
ol-1-yl-phenyl)-4-fluoro-3-(pyridin-3-ylmethoxy)-benzamide;
3-(5-Bromo-pyridin-3-ylmethoxy)-4-fluoro-N-(4-piperazin-1-yl-3-trifluorom-
ethyl-phenyl)-benzamide;
3-(2-Isoquinolin-4-yl-ethyl)-N-phenyl-benzamide;
3-(2-Isoquinolin-4-yl-ethyl)-N-(3,3,5-trimethyl-cyclohexyl)-benzamide;
N-(4-Isopropyl-3-methyl-phenyl)-3-(2-isoquinolin-4-yl-ethyl)-benzamide;
3-(2-Isoquinolin-4-yl-ethyl)-N-(2-methyl-quinolin-6-yl)-benzamide;
N-(3,5-Dibromo-4-methyl-phenyl)-3-(2-isoquinolin-4-yl-ethyl)-benzamide;
N-(4,6-Dimethyl-pyridin-2-yl)-3-(2-isoquinolin-4-yl-ethyl)-benzamide;
2-Chloro-4-fluoro-N-(4-isopropyl-3-methyl-phenyl)-5-(2-isoquinolin-4-yl-e-
thyl)-benzamide;
2,4-Difluoro-N-(4-isopropyl-3-methyl-phenyl)-5-(2-isoquin-
olin-4-yl-ethyl)-benzamide;
2-Fluoro-N-(4-isopropyl-3-methyl-phenyl)-5-(2--
isoquinolin-4-yl-ethyl)-benzamide;
N-(2-Methyl-quinolin-6-yl)-3-(2-pyridin- -3-yl-ethyl)-benzamide
hydrochloride; N-(4-Isopropyl-3-methyl-phenyl)-3-(2-
-pyridin-3-yl-ethyl)-benzamide;
N-(3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-ylsu-
lfanylmethyl]phenyl}-(3-bromo-4-methyl) benzamide;
N-{3-[(1H-pyrazolo[3,4--
d]-pyrimidin-4-ylsulfanylmethyl]phenyl}-3,5-bis(trifluoromethyl)
benzamide;
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl-
}-(4-hydroxy-3-methoxy) benzamide;
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-y-
l)sulfanylmethyl]phenyl}-(4-hydroxy-3-t-butyl) benzamide;
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-4-t-butyl-
benzamide;
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-
-(4-phenoxy)benzamide;
N-{3-[(1H-pyrazolo[3,4-d]pyrimidin-4-ylsulfanyl)met-
hyl]phenyl}-N'-[3,5-bis-(trifluoromethyl)phenyl]urea;
N-{3-[(1H-pyrazolo[3,4-d]pyrimidin-4-ylsulfanyl)methyl]phenyl}-N'-(pyridi-
n-3-yl)urea;
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]pheny-
l}-(3,5-di-t-butyl) benzamide;
3-Bromo-4-hydroxy-N-{3-[(1H-pyrazolo[3,4-d]-
-pyrimidin-4-yl)sulfanylmethyl]phenyl}-benzamide;
N-{3-[(1H-pyrazolo[3,4-d-
]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-quinoline-6-carboxamide;
5-Fluoro-N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}--
indole-2-carboxamide;
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmet-
hyl]phenyl}-indole-6-carboxamide;
(R/S)-2-(2-methylphenyl)-N-{3-[(1H-pyraz-
olo[3,4-d]pyrimidin-4-ylsulfanyl)methyl)-methyl]phenyl}butanamide;
3-t-Butyl-4-hydroxy-N-{3-[5-(6-methoxy-pyridin-3-ylamino)-2H-pyrazol-3-yl-
methyl sulfanyl]-phenyl}-benzamide;
3-t-Butyl-4-hydroxy-N-[3-(pyridin-3-yl-
methylsulfanyl)-phenyl}-benzamide;
3-t-Butyl-4-hydroxy-N-[3-(isoquinolin-4-
-ylmethylsulfanyl)-phenyl}-benzamide;
[3-(5-Bromo-pyridin-3-ylmethoxy)-phe-
nyl]-3-t-butyl-4-hydroxy-benzamide;
4-Acetoxy-3-t-butyl-N-[3-(pyridin-3-yl- methoxy)phenyl]-benzamide;
4-Acetoxy-3-t-butyl-N-[3-(isoquinolin-4-ylmetho-
xy)phenyl]-benzamide;
3-t-Butyl-4-hydroxy-N-[3-(pyridin-3-ylmethoxy)-pheny- l]-benzamide;
3-t-Butyl-4-hydroxy-N-[3-(isoquinolin-4-ylmethoxy)-phenyl]-b-
enzamide; 1-[3-(pyridin-3-ylmethoxy)phenylcarbamoyl]pyrrolidine;
4-[3-(pyridin-3-ylmethoxy)phenylcarbamoyl]morpholine;
3-[{6-Methoxy-7-(2-methoxyethoxy)cinnolin-4-yl}sulfanylmethyl]-N-phenyl-b-
enzamide;
3-[2-(6-Acetylamino-pyridin-3-yl)-ethyl]-N-(4-piperazin-1-yl-3-t-
rifluoro-methylphenyl)-benzamide dihydrochloride;
3-[2-(6-Amino-pyridin-3--
yl)-ethyl]-N-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-benzamide
dihydrochloride;
3-[2-(3H-Imidazo[4,5-b]pyridin-6-yl)-ethyl]-N-(4-piperaz-
in-1-yl-3-trifluoromethyl-phenyl)-benzamide dihydrochloride;
5-{2-[3-(Piperazin-1-yl-trifluoromethyl-phenylcarbamoyl)-phenyl]-ethyl}-n-
icotinamide dihydrochloride;
5-{2-[3-(Piperazin-1-yl-trifluoromethyl-pheny-
lcarbamoyl)-phenyl]-ethyl}-nicotinic acid methyl ester
dihydrochloride;
4-Fluoro-3-[2-(3H-imidazo[4,5-b]pyridin-6-yl)-ethyl]-N-(4-piperazin-1-yl--
3-trifluoromethyl-phenyl)-benzamide dihydrochloride; and
4-Fluoro-3-(5-furan-2-yl-pyridin-3-ylmethoxy)-N-(4-piperazin-1-yl-3-trifl-
uoromethyl-phenyl)-benzamide dihydrochloride; or a pharmaceutically
acceptable salt, pharmaceutically acceptable prodrug, or
pharmaceutically active metabolite thereof or a pharmaceutically
acceptable salt of said metabolite.
13. A compound selected from the group consisting of the compounds
corresponding to Example B-27 (Compounds 1-244), Example V-6d
(Compounds 1-176), Example V-7b (Compounds 1-43) and Example V-14
(Compounds 1-88), or a pharmaceutically acceptable salt,
pharmaceutically acceptable prodrug, or pharmaceutically active
metabolite thereof or a pharmaceutically acceptable salt of said
metabolite.
14. A pharmaceutically acceptable salt of a pharmaceutically active
metabolite of a compound according to claim 1.
15. A pharmaceutical composition for modulating or inhibiting the
activity of a protein kinase receptor comprising: (a) a
therapeutically effective amount of an agent selected from the
group consisting of a compound according to claim 1, a
pharmaceutically acceptable prodrug thereof, a pharmaceutically
active metabolite thereof, and a pharmaceutically acceptable salt
thereof; and (b) a pharmaceutically acceptable carrier, diluent, or
vehicle therefor.
16. A pharmaceutical composition for modulating or inhibiting the
activity of a protein kinase receptor comprising: (a) a
therapeutically effective amount of a pharmaceutically acceptable
salt of a pharmaceutically active metabolite of a compound
according to claim 1; (b) a pharmaceutically acceptable carrier,
diluent, or vehicle therefor.
17. A method of treating a mammalian disease condition mediated by
protein kinase activity, comprising administering to a mammal in
need thereof a therapeutically effective amount of an agent
selected from the group consisting of a compound according to claim
1, a pharmaceutically acceptable prodrug thereof, a
pharmaceutically active metabolite thereof, and a pharmaceutically
acceptable salt thereof.
18. A method according to claim 17, wherein the mammalian disease
condition is associated with tumor growth, cell proliferation, or
angiogenesis.
19. A method of modulating or inhibiting the activity of a protein
kinase receptor, comprising contacting the kinase receptor with an
effective amount of an agent selected from the group consisting of
a compound according to claim 1, a pharmaceutically acceptable
prodrug thereof, a pharmaceutically active metabolite thereof, and
a pharmaceutically acceptable salt thereof.
20. A method according to claim 19, wherein the protein kinase
receptor is a VEGF receptor.
Description
[0001] This applications claims the benefit of U.S. Provisional
Application Serial No. 60/177,059, filed Jan. 21, 2000, the
contents of which are hereby incorporated by reference herein.
FIELD OF THE INVENTION
[0002] This invention is directed to amide compounds that mediate
and/or inhibit the activity of certain protein kinases, and to
pharmaceutical compositions containing such compounds. The
invention is also directed to the therapeutic or prophylactic use
of such compounds and compositions, and to methods of treating
cancer as well as other disease states associated with unwanted
angiogenesis and/or cellular proliferation, by administering
effective amounts of such compounds.
BACKGROUND OF THE INVENTION
[0003] Protein kinases are a family of enzymes that catalyze
phosphorylation of the hydroxyl group of specific tyrosine, serine,
or threonine residues in proteins. Typically, such phosphorylation
dramatically perturbs the function of the protein, and thus protein
kinases are pivotal in the regulation of a wide variety of cellular
processes, including metabolisim, cell proliferation, cell
differentiation, and cell survival. Of the many different cellular
functions in which the activity of protein kinases is known to be
required, some processes represent attractive targets for
therapeutic intervention for certain disease states. Two examples
are angiogenesis and cell-cycle control, in which protein kinases
play a pivotal role; these processes are essential for the growth
of solid tumors as well as for other diseases.
[0004] Angiogenesis is the mechanism by which new capillaries are
formed from existing vessels. When required, the vascular system
has the potential to generate new capillary networks in order to
maintain the proper functioning of tissues and organs. In the
adult, however, angiogenesis is fairly limited, occurring only in
the process of wound healing and neovascularization of the
endometrium during menstruation. See Merenmies, J., Parada, L. F.,
Henkemeyer, M., Cell Growth & Differentiation, 8, 3-10 (1997).
On the other hand, unwanted angiogenesis is a hallmark of several
diseases, such as retinopathies, psoriasis, rheumatoid arthritis,
age-related macular degeneneration, and cancer (solid tumors).
Folkman, Nature Med., 1, 27-31 (1995). Protein kinases which have
been shown to be involved in the angiogenic process include three
members of the growth factor receptor tyrosine kinase family:
VEGF-R2 (vascular endothelial growth factor receptor 2, also known
as KDR (kinase insert domain receptor) and as FLK-1); FGF-R
(fibroblast growth factor receptor); and TEK (also known as
Tie-2).
[0005] VEGF-R2, which is selectively expressed on endothelial
cells, binds the potent angiogenic growth factor VEGF and mediates
the subsequent signal transduction through activation of its
intracellular kinase activity. Thus, it is expected that direct
inhibition of the kinase activity of VEGF-R2 will result in the
reduction of angiogenesis even in the presence of exogenous VEGF
(see Strawn et al., Cancer Research, 56, 3540-3545 (1996)), as has
been shown with mutants of VEGF-R2 which fail to mediate signal
transduction. Millauer et al., Cancer Research, 56, 1615-1620
(1996). Furthermore, VEGF-R2 appears to have no function in the
adult beyond that of mediating the angiogenic activity of VEGF.
Therefore, a selective inhibitor of the kinase activity of VEGF-R2
would be expected to exhibit little toxicity.
[0006] Similarly, FGF-R binds the angiogenic growth factors aFGF
and bFGF and mediates subsequent intracellular signal transduction.
Recently, it has been suggested that growth factors such as bFGF
may play a critical role in inducing angiogenesis in solid tumors
that have reached a certain size. Yoshiji et al., Cancer Research,
57, 3924-3928 (1997). Unlike VEGF-R2, however, FGF-R is expressed
in a number of different cell types throughout the body and may or
may not play important roles in other normal physiological
processes in the adult. Nonetheless, systemic administration of a
small molecule inhibitor of the kinase activity of FGF-R has been
reported to block bFGF-induced angiogenesis in mice without
apparent toxicity. Mohammad et al., EMBO Journal, 17, 5996-5904
(1998).
[0007] TEK (also known as Tie-2) is another receptor tyrosine
kinase selectively expressed on endothelial cells which has been
shown to play a role in angiogenesis. The binding of the factor
angiopoietin-1 results in autophosphorylation of the kinase domain
of TEK and results in a signal transduction process which appears
to mediate the interaction of endothelial cells with
peri-endothelial support cells, thereby facilitating the maturation
of newly formed blood vessels. The factor angiopoietin-2, on the
other hand, appears to antagonize the action of angiopoietin-1 on
TEK and disrupts angiogenesis. Maisonpierre et al., Science, 277,
55-60 (1997).
[0008] As a result of the above-described developments, it has been
proposed to treat angiogenesis by the use of compounds inhibiting
the kinase activity of VEGF-R2, FGF-R, and/or TEK. For example,
WIPO International Publication No. WO 97/34876 discloses certain
cinnoline derivatives that are inhibitors of VEGF-R2, which may be
used for the treatment of disease states associated with abnormal
angiogenesis and/or increased vascular permeability such as cancer,
diabetes, psoriosis, rheumatoid arthritis, Kaposi's sarcoma,
haemangioma, acute and chronic nephropathies, atheroma, arterial
restinosis, autoimmune diseases, acute inflammation and ocular
diseases with retinal vessel proliferation. Two documents described
hereinafter disclose certain amide derivatives but do not disclose
or teach that any of the compounds may be used for modulating or
inhibiting the activity of protein kinases: WIPO International
Publication No. WO 97/03967; and WIPO International Publication No.
WO 96/23783.
[0009] In addition to its role in angiogenesis, protein kinases
also play a crucial role in cell-cycle control. Uncontrolled cell
proliferation is the insignia of cancer. Cell proliferation in
response to various stimuli is manifested by a de-regulation of the
cell division cycle, the process by which cells multiply and
divide. Tumor cells typically have damage to the genes that
directly or indirectly regulate progression through the cell
division cycle.
[0010] Cyclin-dependent kinases (CDKs) are serine-threonine protein
kinases that play critical roles in regulating the transitions
between different phases of the cell-cycle, such as the progression
from a quiescent stage in G.sub.1 (the gap between mitosis and the
onset of DNA replication for a new round of cell division) to S
(the period of active DNA synthesis), or the progression from
G.sub.2 to M phase, in which active mitosis and cell-division
occurs. See, e.g., the articles compiled in Science, 274, 1643-1677
(1996). CDK complexes are formed through association of a
regulatory cyclin subunit (e.g., cyclin A, B 1, B2, D1, D2, D3, and
E) and a catalytic kinase subunit (e.g., cdc2 (CDK1), CDK2, CDK4,
CDK5, and CDK6). As the name implies, the CDKs display an absolute
dependence on the cyclin subunit in order to phosphorylate their
target substrates, and different kinase/cyclin pairs function to
regulate progression through specific phases of the cell-cycle.
[0011] It is CDK4 complexed to the D cyclins that plays a critical
part in initiating the cell-division cycle from a resting or
quiescent stage to one in which cells become committed to cell
division. This progression is subject to a variety of growth
regulatory mechanisms, both negative and positive. Aberrations in
this control system, particularly those that affect the function of
CDK4, have been implicated in the advancement of cells to the
highly proliferative state characteristic of malignancies,
particularly familial melanomas, esophageal carcinomas, and
pancreatic cancers. See, e.g., Hall et al., Adv. Cancer Res., 68,
67-108 (1996); Kamb, Trends in Genetics, 11, 136-140 (1995); Kamb
et al., Science, 264, 436-440 (1994).
[0012] A large number of small molecule ATP-site antagonists have
been identified as CDK inhibitors. (See, Webster, Exp. Opin.
Invest. Drugs, 7, 865-887 (1998), Stover, Et al., Curr. Opin. Drug
Disc. Dev., 2, 274-285(1999), Gray et al., Curr. Med. Chem., 6,
859-875 (1999), Sielecki, et al., J. Med. Chem., 43, 1-18 (2000),
Crews, et al., Curr. Opin. Chem. Biol., 4, 47-53 (2000),
Buolamwini, Curr. Pharm. Des., 6, 379-392 (2000), and Rosania, et
al., Exp. Opin. Ther. Pat., 10, 215-230 (2000)). Moreover, the use
of compounds as anti-proliferative therapeutic agents that inhibit
CDKs is the subject of several patents and publications. For
example, U.S. Pat. No. 5,621,082 to Xiong et al., discloses nucleic
acid encoding an inhibitor of CDK6 and European Patent Publication
No. 0 666 270 A2 describes peptides and peptidemimetics that act as
inhibitors of CDK1 and CDK2. WIPO International Publication No. WO
97/16447 discloses certain analogs of chromones that are inhibitors
of cyclin-dependent kinases, in particular of CDK/cyclin complexes
such as CDK4/cyclin D1, which may be used for inhibiting excessive
or abnormal cell proliferation, and therefore for treating cancer.
WIPO International Publication No. WO 99/21845 describes
4-aminothiazole derivatives that are useful as CDK inhibitors.
[0013] There is still a need, however, for small-molecule compounds
that may be readily synthesized and are effective in inhibiting one
or more CDKs or CDK/cyclin complexes. Because CDK4 may serve as a
general activator of cell division in most cells, and complexes of
CDK4 and D-type cyclins govern the early G.sub.1 phase of the
cell-cycle, there is a need for effective inhibitors of CDK4, and
D-type cyclin complexes thereof, for treating one or more types of
tumors. Also, the pivotal roles of cyclin E/CDK2 and cyclin B/CDK1
kinases in the G1/S phase and G2/M transitions, respectively offer
additional targets for therapeutic intervention in suppressing
deregulated cell-cycle progression in cancer.
[0014] Another protein kinase, CHK-1, plays an important role as a
checkpoint in cell-cycle progression. Checkpoints are control
systems that coordinate cell-cycle progression by influencing the
formation, activation and subsequent inactivation of the
cyclin-dependent kinases. Checkpoints prevent cell-cycle
progression at inappropriate times, maintain the metabolic balance
of cells while the cell is arrested, and in some instances can
induce apoptosis (programmed cell death) when the requirements of
the checkpoint have not been met. See, e.g., O'Connor, Cancer
Surveys, 29, 151-182 (1997); Nurse, Cell, 91, 865-867 (1997);
Hartwell et al., Science, 266, 1821-1828 (1994); Hartwell et al.,
Science, 246, 629-634 (1989).
[0015] One series of checkpoints monitors the integrity of the
genome and, upon sensing DNA damage, these "DNA damage checkpoints"
block cell-cycle progression in G.sub.1 & G.sub.2 phases, and
slow progression through S phase. O'Connor, Cancer Surveys, 29,
151-182 (1997); Hartwell et al., Science, 266, 1821-1828 (1994).
This action enables DNA repair processes to complete their tasks
before replication of the genome and subsequent separation of this
genetic material into new daughter cells takes place. Importantly,
the most commonly mutated gene in human cancer, the p53 tumor
suppressor gene, produces a DNA damage checkpoint protein that
blocks cell-cycle progression in G.sub.1 phase and/or induces
apoptosis (programmed cell death) following DNA damage. Hartwell et
al., Science, 266, 1821-1828 (1994). The p53 tumor suppressor has
also been shown to strengthen the action of a DNA damage checkpoint
in G.sub.2 phase of the cell-cycle. See, e.g., Bunz et al.,
Science, 28, 1497-1501 (1998); Winters et al., Oncogene, 17,
673-684 (1998); Thompson, Oncogene, 15, 3025-3035 (1997).
[0016] Given the pivotal nature of the p53 tumor suppressor pathway
in human cancer, therapeutic interventions that exploit
vulnerabilities in p53-defective cancer have been actively sought.
One emerging vulnerability lies in the operation of the G.sub.2
checkpoint in p53 defective cancer cells. Cancer cells, because
they lack G.sub.1 checkpoint control, are particularly vulnerable
to abrogation of the last remaining barrier protecting them from
the cancer killing effects of DNA-damaging agents: the G.sub.2
checkpoint. The G.sub.2 checkpoint is regulated by a control system
that has been conserved from yeast to humans. Important in this
conserved system is a kinase, CHK-1, which transduces signals from
the DNA-damage sensory complex to inhibit activation of the cyclin
B/Cdc2 kinase, which promotes mitotic entry. See, e.g., Peng et
al., Science, 277, 1501-1505 (1997); Sanchez et al., Science, 277,
1497-1501 (1997). Inactivation of CHK-1 has been shown to both
abrogate G.sub.2 arrest induced by DNA damage inflicted by either
anticancer agents or endogenous DNA damage, as well as result in
preferential killing of the resulting checkpoint defective cells.
See, e.g., Nurse, Cell, 91, 865-867 (1997); Weinert, Science, 277,
1450-1451 (1997); Walworth et al., Nature, 363, 368-371 (1993); and
Al-Khodairy et al., Molec. Biol. Cell, 5, 147-160 (1994).
[0017] Selective manipulation of checkpoint control in cancer cells
could afford broad utilization in cancer chemotherapeutic and
radiotherapy regimens and may, in addition, offer a common hallmark
of human cancer "genomic instability" to be exploited as the
selective basis for the destruction of cancer cells. A number of
factors place CHK-1 as a pivotal target in DNA-damage checkpoint
control. The elucidation of inhibitors of this and functionally
related kinases such as Cds1/CHK-2, a kinase recently discovered to
cooperate with CHK-1 in regulating S phase progression (see Zeng et
al., Nature, 395, 507-510 (1998); Matsuoka, Science, 282, 1893-1897
(1998)), could provide valuable new therapeutic entities for the
treatment of cancer.
[0018] Tyrosine kinases can be of the receptor type (having
extracellular, transmembrane and intracellular domains) or the
non-receptor type (being wholly intracellular). At least one of the
non-receptor protein tyrosine kinases, namely, LCK, is believed to
mediate the transduction in T-cells of a signal from the
interaction of a cell-surface protein (Cd4) with a cross-linked
anti-Cd4 antibody. A more detailed discussion of non-receptor
tyrosine kinases is provided in Bolen, Oncogene, 8, 2025-2031
(1993), which is incorporated herein by reference.
[0019] In addition to the protein kinases identified above, many
other protein kinases have been considered to be therapeutic
targets, and numerous publications disclose inhibitors of kinase
activity, as reviewed in the following: McMahon et al., Current
Opinion in Drug Discovery & Development, 1, 131-146 (1998);
Strawn et al., Exp. Opin. Invest. Drugs, 7, 553-573 (1998); Adams
et al., Curr. Opin. Drug Disc. Dev., 2, 96-109 (1999), Stover et
al., Curr. Opin. Drug Disc. Dev., 2, 274-285 (1999), Toledo et al.,
Curr. Med. Chem., 6, 775-805 (1999), and Garca-Echeverra, et al.,
Med. Res. Rev., 20, 28-57 (2000).
[0020] There is still a need, however, for effective inhibitors of
protein kinases. Moreover, as is well understood by those skilled
in the art, it is desirable for kinase inhibitors to possess both
high affinity for the target kinase as well as high selectivity
versus other protein kinases.
SUMMARY OF THE INVENTION
[0021] Thus, an objective of the invention is to provide potent
inhibitors of protein kinases. Another objective of the invention
is to provide effective kinase inhibitors having a strong and
selective affinity for a particular kinase.
[0022] These and other objectives of the invention, which will
become apparent from the following description, have been achieved
by the discovery of amide compounds, pharmaceutically acceptable
prodrugs, pharmaceutically active metabolites, and pharmaceutically
acceptable salts thereof (such compounds, prodrugs, metabolites and
salts are collectively referred to as "agents") described below,
which modulate and/or inhibit the activity of protein kinases.
[0023] Pharmaceutical compositions containing such agents are
useful in treating various diseases and disorders associated with
uncontrolled or unwanted angiogenesis and/or cellular
proliferation, such as cancer, autoimmune diseases, viral diseases,
fungal diseases, neurodegenerative disorders and cardiovascular
diseases. Thus, pharmaceutical compositions containing such agents
are useful in the treatment of diabetic retinopathy, neovascular
glaucoma, rheumatoid arthritis, and psoriasis.
[0024] Further, the agents have advantageous properties relating to
modulation and/or inhibition of the kinase activity associated with
VEGF-R, FGF-R, CDK complexes (e.g., CDK1, CDK2, CDK4 and CDK6),
CHK-1, TEK, and LCK. Thus, pharmaceutical compositions containing
such agents are useful in the treatment of diseases and disorders
mediated by kinase activity, such as cancer.
[0025] In a general aspect, the invention relates to compounds
represented by the Formula I: 1
[0026] wherein:
[0027] R.sup.1 is a moiety of the formula 2
[0028] where
[0029] Z is CH or NH, and Q is a moiety such that R.sup.1 is a
substituted or unsubstituted monocyclic or bicyclic heteroaryl
which has at least two carbon atoms in the heteroaryl ring
system;
[0030] X is selected from CH.sub.2, O, S, and NH;
[0031] Y is selected from CH.sub.2, O, and S, provided that at
least one of X and Y is CH.sub.2, or X and Y together with the bond
there-between form a cyclopropyl;
[0032] R.sup.2 and R.sup.3 are independently selected from
hydrogen, methyl, halogen, trifluoromethyl, and cyano; and
[0033] R.sup.4 is 3
[0034] where R.sup.5 is a substituted or unsubstituted aryl,
heteroaryl, cycloalkyl, heterocycloalkyl, O--R.sup.7,
NR.sup.8R.sup.9, C.sub.1-C.sub.8 alkyl, or monocyclic
heterocycloalkyl group, R.sup.6 is a substituted or unsubstituted
aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkenyl,
O--R.sup.7, C(O)R.sup.7, NR.sup.8R.sup.9, C.sub.2-C.sub.8 alkyl, or
monocyclic heterocycloalkyl group, where R.sup.7 is a substituted
or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl, R.sup.8 is hydrogen or a substituted or unsubstituted
alkyl, and R.sup.9 is a substituted or unsubstituted alkyl, aryl,
heteroaryl, cycloalkyl, or heterocycloalkyl.
[0035] The invention is also directed to pharmaceutically
acceptable prodrugs, pharmaceutically active metabolites, and
pharmaceutically acceptable salts of the compounds of Formula I.
Pharmaceutically acceptable salts of such active metabolites are
also provided. Advantageous methods of making the compounds of the
Formula I are also described.
[0036] In a preferred general embodiment, the invention relates to
compounds of the Formula I wherein R.sup.1 is a substituted or
unsubstituted heteroaryl group selected from: 4
[0037] where
[0038] X is selected from CH.sub.2, O, and S;
[0039] Y is selected from CH.sub.2 and S, provided that at least
one of X and Y is CH.sub.2;
[0040] R.sup.2 and R.sup.3 are independently selected from
hydrogen, methyl, fluorine, and chlorine; and
[0041] R.sup.4 is 5
[0042] where R.sup.5 is a substituted or unsubstituted aryl,
heteroaryl, cycloalkyl, heterocycloalkyl, O--R.sup.7,
NR.sup.8R.sup.9, C.sub.1-C.sub.8 alkyl, or monocyclic
heterocycloalkyl group, R.sup.6 is a substituted or unsubstituted
aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkenyl,
O--R.sup.7, C(O)R.sup.7, NR.sup.8R.sup.9, C.sub.2-C.sub.8 alkyl, or
monocyclic heterocycloalkyl group, where R.sup.7 is a substituted
or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl, R.sup.8 is hydrogen or a substituted or unsubstituted
alkyl, and R.sup.9 is a substituted or unsubstituted alkyl, aryl,
heteroaryl, cycloalkyl, or heterocycloalkyl.
[0043] Especially preferred are compounds represented by the
Formula II: 6
[0044] wherein:
[0045] X is selected from CH.sub.2, O, and S;
[0046] Y is selected from CH.sub.2 and S, provided that at least
one of X and Y is CH.sub.2;
[0047] R.sup.2 and R.sup.3 are independently selected from
hydrogen, methyl, fluorine, and chlorine;
[0048] R.sup.4 is 7
[0049] where R.sup.5 and R.sup.6 are each independently a
substituted or unsubstituted aryl or heteroaryl; and R.sup.10 is a
substituted or unsubstituted alkenyl, aryl, heteroaryl, or
NHR.sup.9, where R.sup.9 is a substituted or unsubstituted alkyl,
aryl, heteroaryl, cycloalkyl, or heterocycloalkyl.
[0050] In another embodiment, the present invention is directed to
compounds represented by the Formula III: 8
[0051] wherein:
[0052] X is selected from CH.sub.2, O, S, and NH;
[0053] Y is selected from CH.sub.2, O, and S, provided that at
least one of X and Y is CH.sub.2, or X and Y together with the bond
there-between form a cyclopropyl;
[0054] R.sup.2 and R.sup.3 are independently selected from
hydrogen, methyl, halogen, trifluoromethyl, and cyano; and
[0055] R.sup.4 is 9
[0056] where R.sup.5 is a substituted or unsubstituted aryl,
heteroaryl, cycloalkyl, heterocycloalkyl, O--R.sup.7,
NR.sup.8R.sup.9, C.sub.1-C.sub.8 alkyl, or monocyclic
heterocycloalkyl group, R.sup.6 is a substituted or unsubstituted
aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkenyl,
O--R.sup.7, C(O)R.sup.7, NR.sup.8R.sup.9, C.sub.2-C.sub.8 alkyl, or
monocyclic heterocycloalkyl group, where R.sup.7 is a substituted
or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl, R.sup.8 is hydrogen or a substituted or unsubstituted
alkyl, and R.sup.9 is a substituted or unsubstituted alkyl, aryl,
heteroaryl, cycloalkyl, or heterocycloalkyl; and pharmaceutically
acceptable salts thereof and pharmaceutically acceptable prodrugs
thereof.
[0057] In preferred embodiments of compounds of Formula III:
[0058] X is selected from CH.sub.2, O, and S;
[0059] Y is selected from CH.sub.2 and S, provided that at least
one of X and Y is CH.sub.2;
[0060] R.sup.2 and R.sup.3 are independently selected from
hydrogen, methyl, fluorine, and chlorine; and
[0061] R.sup.4 is 10
[0062] where R.sup.5 and R.sup.6 are each independently a
substituted or unsubstituted aryl or heteroaryl.
[0063] Especially preferred are compounds represented by the
Formula III, wherein:
[0064] X is CH.sub.2;
[0065] Y is S;
[0066] R.sup.2 and R.sup.3 are independently selected from
hydrogen, methyl, fluorine, and chlorine; and
[0067] R.sup.4 is 11
[0068] where R.sup.5 and R.sup.6 are each independently a
substituted or unsubstituted aryl or heteroaryl.
[0069] The invention also relates to a method of modulating and/or
inhibiting the kinase activity of VEGF-R, FGF-R, a CDK complex,
CHK-1, TEK, and/or LCK by administering a compound of the Formula
I, II, or III, or a pharmaceutically acceptable prodrug,
pharmaceutically active metabolites, or pharmaceutically acceptable
salt thereof. Preferably, compounds of the present invention have
selective kinase activity--i.e., they possess significant activity
against one specific kinase while possessing less or minimal
activity against a different kinase. In one preferred embodiment of
the invention, compounds of the present invention are those of
Formula I, II, or III, possessing substantially higher potency
against VEGF receptor tyrosine kinase than against LCK receptor
tyrosine kinase. The invention is also directed to methods of
modulating VEGF receptor tyrosine kinase activity without
significantly modulating LCK receptor tyrosine kinase activity.
[0070] The invention also relates to pharmaceutical compositions
each comprising: an effective amount of an agent selected from
compounds of the Formula I, II, and III, and pharmaceutically
acceptable salts, pharmaceutically active metabolites, and
pharmaceutically acceptable prodrugs thereof; and a
pharmaceutically acceptable carrier or vehicle for such agent.
[0071] The invention further provides methods of treating cancer as
well as other disease states associated with unwanted angiogenesis
and/or cellular proliferation, comprising administering effective
amounts of such agents to a patient in need of such treatment.
DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS
[0072] The inventive compounds of the Formula I, II, and III are
useful for mediating the activity of protein kinases. More
particularly, the compounds are useful as anti-angiogenesis agents
and as agents for modulating and/or inhibiting the activity of
protein kinases, thus providing treatments for cancer or other
diseases associated with cellular proliferation mediated by protein
kinases.
[0073] The term "alkyl" as used herein refers to straight- and
branched-chain alkyl groups having one to twelve carbon atoms.
Exemplary alkyl groups include methyl (Me), ethyl, n-propyl,
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (tBu), pentyl,
isopentyl, tert-pentyl, hexyl, isohexyl, and the like. The term
"alkenyl" refers to straight- and branched-chain alkenyl groups
having from two to twelve carbon atoms. Illustrative alkenyl groups
include prop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl,
hex-2-enyl, and the like.
[0074] The term "cycloalkyl" refers to saturated or unsaturated
carbocycles having from three to twelve carbon atoms, including
bicyclic and tricyclic cycloalkyl structures. Suitable cycloalkyls
include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, and the like.
[0075] A "heterocycloalkyl" group is intended to mean a saturated
or unsaturated monocyclic radical containing carbon atoms,
preferably 4 or 5 ring carbon atoms, and at least one heteroatom
selected from nitrogen, oxygen and sulfur.
[0076] The terms "aryl" (Ar) and "heteroaryl" refer to monocyclic
and polycyclic unsaturated or aromatic ring structures, with "aryl"
referring to those that are carbocycles and "heteroaryl" referring
to those that are heterocycles. Examples of aromatic ring
structures include phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl,
furyl, thienyl, pyrrolyl, pyridyl, pyridinyl, pyrazolyl,
imidazolyl, pyrazinyl, pyridazinyl, 1,2,3-triazinyl,
1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1-H-tetrazol-5-yl, indolyl,
quinolinyl, benzofuranyl, benzothiophenyl (thianaphthenyl), and the
like. Such moieties may be optionally substituted by one or more
suitable substituents, for example, a substituent selected from a
halogen (F, Cl, Br or I); lower alkyl; OH; NO.sub.2; CN; CO.sub.2H;
O-lower alkyl; aryl; aryl-lower alkyl; CO.sub.2CH.sub.3;
CONH.sub.2; OCH.sub.2CONH.sub.2; NH.sub.2; SO.sub.2NH.sub.2;
OCHF.sub.2; CF.sub.3; OCF.sub.3; and the like. Such moieties may
also be optionally substituted by a fused-ring structure or bridge,
for example OCH.sub.2--O.
[0077] The term "alkoxy" is intended to mean the radical --O-alkyl.
Illustrative examples include methoxy, ethoxy, propoxy, and the
like.
[0078] The term "halogen" represents chlorine, fluorine, bromine or
iodine. The term "halo" represents chloro, fluoro, bromo or
iodo.
[0079] In general, the various moieties or functional groups for
variables in the formulae may be optionally substituted by one or
more suitable substituents. Exemplary substituents include a
halogen (F, Cl, Br, or I), lower alkyl, --OH, --NO.sub.2, --CN,
--CO.sub.2H, --O-lower alkyl, -aryl, -aryl-lower alkyl,
--CO.sub.2CH.sub.3, --CONH.sub.2, --OCH.sub.2CONH.sub.2,
--NH.sub.2, --SO.sub.2NH.sub.2, haloalkyl (e.g., --CF.sub.3,
--CH.sub.2CF.sub.3), --O-haloalkyl (e.g., --OCF.sub.3,
--OCHF.sub.2), and the like.
[0080] It is understood that compounds of Formula I, II, and III
may exhibit the phenomenon of tautomerism and that the formula
drawings within this specification can represent only one of the
possible tautomeric forms. It is to be understood that the
invention encompasses any tautomeric form which modulates and/or
inhibits kinase activity and is not to be limited merely to any one
tautomeric form utilized within the formula drawings.
[0081] Some of the inventive compounds may exist as single
stereoisomers (i.e., essentially free of other stereoisomers),
racemates, and/or mixtures of enantiomers and/or diastereomers. All
such single stereoisomers, racemates and mixtures thereof are
intended to be within the scope of the present invention.
Preferably, the inventive compounds that are optically active are
used in optically pure form.
[0082] As generally understood by those skilled in the art, an
optically pure compound having one chiral center (i.e., one
asymmetric carbon atom) is one that consists essentially of one of
the two possible enantiomers (i.e., is enantiomerically pure), and
an optically pure compound having more than one chiral center is
one that is both diastereomerically pure and enantiomerically pure.
Preferably, the compounds of the present invention are used in a
form that is at least 90% optically pure, that is, a form that
contains at least 90% of a single isomer (80% enantiomeric excess
("e.e.") or diastereomeric excess ("d.e.")), more preferably at
least 95% (90% e.e. or d.e.), even more preferably at least 97.5%
(95% e.e. or d.e.), and most preferably at least 99% (98% e.e. or
d.e.).
[0083] Additionally, the formulas are intended to cover solvated as
well as unsolvated forms of the identified structures. For example,
Formula I includes compounds of the indicated structure in both
hydrated and non-hydrated forms. Other examples of solvates include
the structures in combination with isopropanol, ethanol, methanol,
DMSO, ethyl acetate, acetic acid, or ethanolamine.
[0084] In addition to compounds of the Formula I, II, and III, the
invention includes pharmaceutically acceptable prodrugs,
pharmaceutically active metabolites, and pharmaceutically
acceptable salts of such compounds. Pharmaceutically active salts
of such active metabolites are also included.
[0085] The term "pharmaceutically acceptable" means
pharmacologically acceptable and substantially non-toxic to the
subject being administered the cell-cycle control agent.
[0086] "A pharmaceutically acceptable prodrug" is a compound that
may be converted under physiological conditions or by solvolysis to
the specified compound or to a pharmaceutically acceptable salt of
such compound.
[0087] "A pharmaceutically active metabolite" is intended to mean a
pharmacologically active product produced through metabolism in the
body of a specified compound or salt thereof. Metabolites of a
compound may be identified using routine techniques known in the
art and their activities determined using tests such as those
described herein.
[0088] Prodrugs and active metabolites of a compound may be
identified using routine techniques known in the art. See, e.g.,
Bertolini et al., J. Med. Chem., 40, 2011-2016 (1997); Shan, et
al., J. Pharm. Sci., 86 (7), 765-767; Bagshawe, Drug Dev. Res., 34,
220-230 (1995); Bodor, Advances in Drug Res., 13, 224-331 (1984);
Bundgaard, Design of Prodrugs (Elsevier Press 1985); and Larsen,
Design and Application of Prodrugs, Drug Design and Development
(Krogsgaard-Larsen et al., eds., Harwood Academic Publishers,
1991).
[0089] "A pharmaceutically acceptable salt" is intended to mean a
salt that retains the biological effectiveness of the free acids
and bases of the specified compound and that is not biologically or
otherwise undesirable. A compound of the invention may possess a
sufficiently acidic, a sufficiently basic, or both functional
groups, and accordingly react with any of a number of inorganic or
organic bases, and inorganic and organic acids, to form a
pharmaceutically acceptable salt. Exemplary pharmaceutically
acceptable salts include those salts prepared by reaction of the
compounds of the present invention with a mineral or organic acid
or an inorganic base, such as salts including sulfates,
pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,
monohydrogenphosphates, dihydrogenphosphates, metaphosphates,
pyrophosphates, chlorides, bromides, iodides, acetates,
propionates, decanoates, caprylates, acrylates, formates,
isobutyrates, caproates, heptanoates, propiolates, oxalates,
malonates, succinates, suberates, sebacates, fumarates, maleates,
butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates,
methylbenzoates, dinitrobenzoates, hydroxybenzoates,
methoxybenzoates, phthalates, sulfonates, xylenesulfonates,
phenylacetates, phenylpropionates, phenylbutyrates, citrates,
lactates, .gamma.-hydroxybutyrates, glycollates, tartrates,
methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,
naphthalene-2-sulfonates, and mandelates.
[0090] If the inventive compound is a base, the desired
pharmaceutically acceptable salt may be prepared by any suitable
method available in the art, for example, treatment of the free
base with an inorganic acid, such as hydrochloric acid, hydrobromic
acid, sulfuric acid, nitric acid, phosphoric acid and the like, or
with an organic acid, such as acetic acid, maleic acid, succinic
acid, mandelic acid, fumaric acid, malonic acid, pyrovic acid,
oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid,
such as glucuronic acid or galacturonic acid, an alpha-hydrozy
acid, such as citric acid or tartaric acid, an amino acid, such as
aspartic acid or glutamic acid, an aromatic acid, such as benzoic
acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic
acid or ethanesulfonic acid, or the like.
[0091] If the inventive compound is an acid, the desired
pharmaceutically acceptable salt may be prepared by any suitable
method, for example, treatment of the free acid with an inorganic
or organic base, such as an amine (primary, secondary or tertiary),
an alkali metal hydroxide or alkaline earth metal hydroxide, or the
like. Illustrative examples of suitable salts include organic salts
derived from amino acids, such as glycine and arginine, ammonia,
primary, secondary, and tertiary amines, and cyclic amines, such as
piperidine, morpholine and piperazine, and inorganic salts derived
from sodium, calcium, potassium, magnesium, manganese, iron,
copper, zinc, aluminum and lithium.
[0092] In the case of agents that are solids, it is understood by
those skilled in the art that the inventive compounds and salts may
exist in different crystal or polymorphic forms, all of which are
intended to be within the scope of the present invention and
specified formulas.
[0093] Therapeutically effective amounts of the agents of the
invention may be used to treat diseases mediated by modulation or
regulation of protein kinases. An "effective amount" is intended to
mean that amount of an agent that, when administered to a mammal in
need of such treatment, is sufficient to effect treatment for a
disease mediated by the activity of one or more protein kinases,
such as tyrosine kinases. Thus, e.g., a therapeutically effective
amount of a compound of the Formula I, salt, active metabolite or
prodrug thereof is a quantity sufficient to modulate, regulate, or
inhibit the activity of one or more protein kinases such that a
disease condition which is mediated by that activity is reduced or
alleviated.
[0094] The amount of a given agent that will correspond to such an
amount will vary depending upon factors such as the particular
compound, disease condition and its severity, the identity (e.g.,
weight) of the mammal in need of treatment, but can nevertheless be
routinely determined by one skilled in the art. "Treating" is
intended to mean at least the mitigation of a disease condition in
a mammal, such as a human, that is affected, at least in part, by
the activity of one or more protein kinases, such as tyrosine
kinases, and includes: preventing the disease condition from
occurring in a mammal, particularly when the mammal is found to be
predisposed to having the disease condition but has not yet been
diagnosed as having it; modulating and/or inhibiting the disease
condition; and/or alleviating the disease condition.
[0095] The inventive agents may be prepared using the reaction
routes and synthesis schemes as described below, employing the
techniques available in the art using starting materials that are
readily available.
[0096] Compounds of formula I where R.sup.4 is CONHR.sup.5 may be
prepared as shown in Scheme 1. 12
[0097] As shown in Scheme 1, carboxylic acids of formula IV are
coupled to amines of formula V to give compounds of formula I
(R.sup.4.dbd.CONHR.sup.5). The coupling may be carried out
employing various peptide coupling reagents, for example
1-ethyl-3-(3'-dimethylamin- opropyl)carbodiimide HCl (EDC),
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetra- methyluronium
hexafluorophosphate (HATU), or benzotriazole-1-yl-oxy-tris-p-
yrrolidino-phosphonium hexafluorophosphate (pyBop), in polar
aprotic solvents, such as N, N-dimethylforamide (DMF) or
dichloromethane. Alternatively, the acid IV may be first converted
to an acid chloride by treatment with, for example, oxalyl chloride
or thionyl chloride, and then, without purification, reacted with
amines of formula V to give compounds of formula I
(R.sup.4.dbd.CONHR.sup.5).
[0098] Compounds of formula I where R.sup.4 is NHCOR.sup.6 may be
prepared as shown in Scheme 2. 13
[0099] Carboxylic acids of formula VII are coupled to amines of
formula VI to give compounds of formula I
(R.sup.4.dbd.NHCOR.sup.6). The coupling may be carried out
employing with various peptide coupling reagents, for example EDC,
HATU, or pyBOP, in polar aprotic solvents, such as DMF or
dichloromethane. Alternatively, the acid VII may be first converted
to an acid chloride by treatment with, for example, oxalyl chloride
or thionyl chloride, and then, without purification, reacted with
amines of formula VI to give compounds of formula I
(R.sup.4.dbd.NHCOR.sup.6).
[0100] Compounds of formula I-a may be prepared by the reaction
shown in Scheme 3. 14
[0101] Compounds of formula IX (R.dbd.R.sup.4) are treated with
compounds of formula VIII, where Lg is a suitable leaving group
such as chloride, bromide, or mesylate, in a dipolar aprotic
solvent such as acetone, DMF, or DMSO, in the presence of a
suitable base, such as potassium carbonate, cesium carbonate,
sodium hydride, and the like, to provide, after extractive workup
and conventional purification, compounds of formula I-a.
Alternatively, this reaction may be carried out in the same manner
with compounds of formula XI (R.dbd.CO.sub.2R.sup.11, where
R.sup.11 is hydrogen or a suitable carboxylic acid protecting
group, such as methyl, ethyl, or benzyl) to give compounds of
formula IV-a. In addition, the reaction carried out with compounds
of formula IX (R.dbd.NH.sub.2) provides compounds of formula
VI-a.
[0102] Compounds of formula I-b may be prepared by the reaction
shown in Scheme 4. 15
[0103] Compounds of formula XI (R.dbd.R.sup.4), where Lg is a
suitable leaving group such as chloride, bromide, or mesylate, are
treated with compounds of formula X in a dipolar aprotic solvent
such as acetone, DMF, or DMSO, in the presence of a suitable base,
such as potassium carbonate, cesium carbonate, sodium hydride, and
the like, to provide, after extractive workup and conventional
purification, compounds of formula I-b. Alternatively, this
reaction may be carried out in the same manner with compounds of
formula XI (R.dbd.CO.sub.2R.sup.11, where R.sup.11 is hydrogen or a
suitable carboxylic acid protecting group, such as methyl, ethyl,
or benzyl) to give compounds of formula IV-b. In addition, the
reaction carried out with compounds of formula XI (R.dbd.NH.sub.2)
provides compounds of formula VI-b.
[0104] Compounds of formula I-c may be prepared by the reaction
shown in Scheme 5. 16
[0105] Compounds of formula XII, where W is a suitable group that
can participate in a palladium-catalyzed coupling reaction such as
bromide, iodide, or triflate, are allowed to react with acetylenes
of formula XIII in the presence of a suitable palladium catalyst,
such as dichlorobis(triphenylphosphine)palladium, and a copper
catalyst, such as cuprous iodide, in the presence of a suitable
base, such as piperidine, triethylaamine or diisopropylethylamine,
in an aprotic solvent, such as THF or DMF, at a temperature between
25.degree. C. and 125.degree. C., for 1 to 24 hours. After
extractive work-up and conventional purification, removal of the
silyl protecting group is effected with, for example, either
tetrabutylammonium fluoride in THF or sodium hydroxide in methanol,
to provide compounds of formula XIV.
[0106] Under similar catalyzed coupling conditions as those
described above, compounds of formula XV can be reacted with those
of formula XIV to yield compounds of formula XVI. Catalytic
hydrogenation of alkynes of formula XVI provides compounds of
formula I-c after filtration and convention purification. Typical
catalytic conditions include catalysts such as palladium, rhodium,
preferably palladium-on-carbon, in a suitable solvent such as
C.sub.1-C.sub.4 alcohols, preferably ethanol.
[0107] Compounds of formula II-a may be prepared as shown in Scheme
6. 17
[0108] Thioamides of formula XVII are treated with two molar
equivalents of a suitable strong base, such as n-butyllithium or
lithium diisopropylamide, in a suitable solvent, such as THF, at
-78.degree. C. to 0.degree. C., to give a solution of thioamide
dianion, which is further treated with less than or equal to 0.5
molar equivalents of compounds of formula XVIII. Conventional
aqueous work-up and purification then provides compounds of formula
XIX, which upon treatment with hydrazine, preferably in the
presence of acetic acid, in ethanol at 0.degree. C. to 50.degree.
C., preferably at room temperature, provides compounds of formula
II-a.
[0109] Compounds of formula II-b may be prepared as shown in Scheme
7. 18
[0110] Ketones of formula XX, where R.sup.12 is substituted or
unsubstituted alkenyl, aryl, or heteroaryl, are converted to
hydrazones of formula XXI by treatment with t-butyl carbazate and
acetic acid in ethanol. Treatment of hydrazones of formula XXI with
two molar equivalents of a suitable strong base, such as
n-butyllithium or lithium diisopropylamide, in a suitable solvent,
such as THF, at -78.degree. C. to 0.degree. C., generates a
solution of hydrazone dianion, which is further treated with less
than or equal to 0.5 molar equivalents of compounds of formula
XVIII. Conventional aqueous work-up and purification then provides
compounds of formula XXII, which upon further treatment with a
suitable acid, such as trifluoroacetic acid, provides compounds of
formula II-b.
[0111] Compounds of formula XVIII are prepared as shown in Scheme
8. 19
[0112] Thus, compounds of formula XVIII-a and XVIII-b are prepared
by alkylation of compounds of formula IX with N-methoxy-N-methyl
chloroacetamide in a like manner to that shown in Scheme 3 above.
Compounds of formula XVIII-c may be prepared by (1) reaction of
aldehydes of formula XXIII with the anion derived from
N-methoxy-N-methyl triethylphosphonoacetamide to give unsaturated
amides of formula XXIV, and (2) reduction of compounds of formula
XXIV with, for example, hydrogen in the presence of palladium on
carbon to provide compounds of formula XVIII-c.
[0113] Other compounds of Formula I, II, and III may be prepared in
manners analogous to the general procedures described above or the
detailed procedures described in the examples herein.
[0114] The affinity of the compounds of the invention for a
receptor may be enhanced by providing multiple copies of the ligand
in close proximity, preferably using a scaffolding provided by a
carrier moiety. It has been shown that provision of such multiple
valence compounds with optimal spacing between the moieties
dramatically improves binding to a receptor. See e.g., Lee et al.,
Biochem, 23, 4255 (1984). The multivalency and spacing can be
controlled by selection of a suitable carrier moiety or linker
units. Such moieties include molecular supports which contain a
multiplicity of functional groups that can be reacted with
functional groups associated with the compounds of the invention.
Of course, a variety of carriers can be used, including proteins
such as BSA or HAS, a multiplicity of peptides including, for
example, pentapeptides, decapeptides, pentadecapeptides, and the
like. The peptides or proteins can contain the desired number of
amino acid residues having free amino groups in their side chains;
however, other functional groups, such as mercapto (--SH) groups or
hydroxyl (--OH) groups, can also be used to obtain stable
linkages.
[0115] Compounds that potently regulate, modulate, or inhibit the
protein kinase activity associated with receptors VEGF, FGF, CDK
complexes, TEK, CHK-1, and LCK, among others, and which inhibit
angiogenesis and/or cellular profileration is desirable and is one
preferred embodiment of the present invention. The present
invention is further directed to methods of modulating or
inhibiting protein kinase activity, for example in mammalian
tissue, by administering an inventive agent. The activity of the
inventive compounds as modulators of protein kinase activity, such
as the activity of kinases, may be measured by any of the methods
available to those skilled in the art, including in vivo and/or in
vitro assays. Examples of suitable assays for activity measurements
include those described in Parast C. et al., BioChemistry, 37,
16788-16801 (1998); Jeffrey et al., Nature, 376, 313-320 (1995);
WIPO International Publication No. WO 97/34876; and WIPO
International Publication No. WO 96/14843. These properties may be
assessed, for example, by using one or more of the biological
testing procedures set out in the examples below.
[0116] The active agents of the invention may be formulated into
pharmaceutical compositions as described below. Pharmaceutical
compositions of this invention comprise an effective modulating,
regulating, or inhibiting amount of a compound of Formula I, II, or
III and an inert, pharmaceutically acceptable carrier or diluent.
In one embodiment of the pharmaceutical compositions, efficacious
levels of the inventive agents are provided so as to provide
therapeutic benefits involving modulation of protein kinases. By
"efficacious levels" is meant levels in which the effects of
protein kinases are, at a minimum, regulated. These compositions
are prepared in unit-dosage form appropriate for the mode of
administration, e.g., parenteral or oral administration.
[0117] An inventive agent is administered in conventional dosage
form prepared by combining a therapeutically effective amount of an
agent (e.g., a compound of Formula I) as an active ingredient with
appropriate pharmaceutical carriers or diluents according to
conventional procedures. These procedures may involve mixing,
granulating and compressing or dissolving the ingredients as
appropriate to the desired preparation.
[0118] The pharmaceutical carrier employed may be either a solid or
liquid. Exemplary of solid carriers are lactose, sucrose, talc,
gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and
the like. Exemplary of liquid carriers are syrup, peanut oil, olive
oil, water and the like. Similarly, the carrier or diluent may
include time-delay or time-release material known in the art, such
as glyceryl monostearate or glyceryl distearate alone or with a
wax, ethylcellulose, hydroxypropylmethylcellulose,
methylmethacrylate and the like.
[0119] A variety of pharmaceutical forms can be employed. Thus, if
a solid carrier is used, the preparation can be tableted, placed in
a hard gelatin capsule in powder or pellet form or in the form of a
troche or lozenge. The amount of solid carrier may vary, but
generally will be from about 25 mg to about 1 g. If a liquid
carrier is used, the preparation will be in the form of syrup,
emulsion, soft gelatin capsule, sterile injectable solution or
suspension in an ampule or vial or non-aqueous liquid
suspension.
[0120] To obtain a stable water-soluble dose form, a
pharmaceutically acceptable salt of an inventive agent is dissolved
in an aqueous solution of an organic or inorganic acid, such as
0.3M solution of succinic acid or citric acid. If a soluble salt
form is not available, the agent may be dissolved in a suitable
cosolvent or combinations of cosolvents. Examples of suitable
cosolvents include, but are not limited to, alcohol, propylene
glycol, polyethylene glycol 300, polysorbate 80, gylcerin and the
like in concentrations ranging from 0-60% of the total volume. In
an exemplary embodiment, a compound of Formula I is dissolved in
DMSO and diluted with water. The composition may also be in the
form of a solution of a salt form of the active ingredient in an
appropriate aqueous vehicle such as water or isotonic saline or
dextrose solution.
[0121] It will be appreciated that the actual dosages of the agents
used in the compositions of this invention will vary according to
the particular complex being used, the particular composition
formulated, the mode of administration and the particular site,
host and disease being treated. Optimal dosages for a given set of
conditions can be ascertained by those skilled in the art using
conventional dosage-determination tests in view of the experimental
data for an agent. For oral administration, an exemplary daily dose
generally employed is from about 0.001 to about 1000 mg/kg of body
weight, more preferably from about 0.001 to about 50 mg/kg body
weight, with courses of treatment repeated at appropriate
intervals. Administration of prodrugs are typically dosed at weight
levels which are chemically equivalent to the weight levels of the
fully active form.
[0122] The compositions of the invention may be manufactured in
manners generally known for preparing pharmaceutical compositions,
e.g., using conventional techniques such as mixing, dissolving,
granulating, dragee-making, levigating, emulsifying, encapsulating,
entrapping or lyophilizing. Pharmaceutical compositions may be
formulated in a conventional manner using one or more
physiologically acceptable carriers, which may be selected from
excipients and auxiliaries that facilitate processing of the active
compounds into preparations which can be used pharmaceutically.
[0123] Proper formulation is dependent upon the route of
administration chosen. For injection, the agents of the invention
may be formulated into aqueous solutions, preferably in
physiologically compatible buffers such as Hanks's solution,
Ringer's solution, or physiological saline buffer. For transmucosal
administration, penetrants appropriate to the barrier to be
permeated are used in the formulation. Such penetrants are
generally known in the art.
[0124] For oral administration, the compounds can be formulated
readily by combining the active compounds with pharmaceutically
acceptable carriers known in the art. Such carriers enable the
compounds of the invention to be formulated as tablets, pills,
dragees, capsules, liquids, gels, syrups, slurries, suspensions and
the like, for oral ingestion by a patient to be treated.
Pharmaceutical preparations for oral use can be obtained using a
solid excipient in admixture with the active ingredient (agent),
optionally grinding the resulting mixture, and processing the
mixture of granules after adding suitable auxiliaries, if desired,
to obtain tablets or dragee cores. Suitable excipients include:
fillers such as sugars, including lactose, sucrose, mannitol, or
sorbitol; and cellulose preparations, for example, maize starch,
wheat starch, rice starch, potato starch, gelatin, gum, methyl
cellulose, hydroxypropylmethyl-cellulose, sodium
carboxymethylcellulose, or polyvinylpyrrolidone (PVP). If desired,
disintegrating agents may be added, such as crosslinked polyvinyl
pyrrolidone, agar, or alginic acid or a salt thereof such as sodium
alginate.
[0125] Dragee cores are provided with suitable coatings. For this
purpose, concentrated sugar solutions may be used, which may
optionally contain gum arabic, polyvinyl pyrrolidone, Carbopol gel,
polyethylene glycol, and/or titanium dioxide, lacquer solutions,
and suitable organic solvents or solvent mixtures. Dyestuffs or
pigments may be added to the tablets or dragee coatings for
identification or to characterize different combinations of active
agents.
[0126] Pharmaceutical preparations which can be used orally include
push-fit capsules made of gelatin, as well as soft, sealed capsules
made of gelatin and a plasticizer, such as glycerol or sorbitol.
The push-fit capsules can contain the active ingredients in
admixture with fillers such as lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate, and,
optionally, stabilizers. In soft capsules, the active agents may be
dissolved or suspended in suitable liquids, such as fatty oils,
liquid paraffin, or liquid polyethylene glycols. In addition,
stabilizers may be added. All formulations for oral administration
should be in dosages suitable for such administration. For buccal
administration, the compositions may take the form of tablets or
lozenges formulated in conventional manner.
[0127] For administration intranasally or by inhalation, the
compounds for use according to the present invention are
conveniently delivered in the form of an aerosol spray presentation
from pressurized packs or a nebuliser, with the use of a suitable
propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethan- e, carbon dioxide or other suitable gas.
In the case of a pressurized aerosol the dosage unit may be
determined by providing a valve to deliver a metered amount.
Capsules and cartridges of gelatin for use in an inhaler or
insufflator and the like may be formulated containing a powder mix
of the compound and a suitable powder base such as lactose or
starch.
[0128] The compounds may be formulated for parenteral
administration by injection, e.g., by bolus injection or continuous
infusion. Formulations for injection may be presented in
unit-dosage form, e.g., in ampoules or in multi-dose containers,
with an added preservative. The compositions may take such forms as
suspensions, solutions or emulsions in oily or aqueous vehicles,
and may contain formulatory agents such as suspending, stabilizing
and/or dispersing agents.
[0129] Pharmaceutical formulations for parenteral administration
include aqueous solutions of the active compounds in water-soluble
form. Additionally, suspensions of the active agents may be
prepared as appropriate oily injection suspensions. Suitable
lipophilic solvents or vehicles include fatty oils such as sesame
oil, or synthetic fatty acid esters, such as ethyl oleate or
triglycerides, or liposomes. Aqueous injection suspensions may
contain substances which increase the viscosity of the suspension,
such as sodium carboxymethyl cellulose, sorbitol, or dextran.
Optionally, the suspension may also contain suitable stabilizers or
agents which increase the solubility of the compounds to allow for
the preparation of highly concentrated solutions.
[0130] For administration to the eye, a compound of the formula I,
II, or m is delivered in a pharmaceutically acceptable ophthalmic
vehicle such that the compound is maintained in contact with the
ocular surface for a sufficient time period to allow the compound
to penetrate the corneal and internal regions of the eye,
including, for example, the anterior chamber, posterior chamber,
vitreous body, aqueous humor, vitreous humor, cornea, iris/cilary,
lens, choroid/retina and selera. The pharmaceutically acceptable
ophthalmic vehicle may be an ointment, vegetable oil, or an
encapsulating material. A compound of the invention may also be
injected directly into the vitreous and aqueous humor.
[0131] Alternatively, the active ingredient may be in powder form
for constitution with a suitable vehicle, e.g., sterile
pyrogen-free water, before use. The compounds may also be
formulated in rectal compositions such as suppositories or
retention enemas, e.g, containing conventional suppository bases
such as cocoa butter or other glycerides.
[0132] In addition to the formulations described above, the
compounds may also be formulated as a depot preparation. Such
long-acting formulations may be administered by implantation (for
example, subcutaneously or intramuscularly) or by intramuscular
injection. Thus, for example, the compounds may be formulated with
suitable polymeric or hydrophobic materials (for example, as an
emulsion in an acceptable oil) or ion-exchange resins, or as
sparingly soluble derivatives, for example, as a sparingly soluble
salt.
[0133] A pharmaceutical carrier for hydrophobic compounds is a
cosolvent system comprising benzyl alcohol, a nonpolar surfactant,
a water-miscible organic polymer, and an aqueous phase. The
cosolvent system may be a VPD co-solvent system. VPD is a solution
of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant
polysorbate 80, and 65% w/v polyethylene glycol 300, made up to
volume in absolute ethanol. The VPD co-solvent system (VPD:5W)
contains VPD diluted 1:1 with a 5% dextrose in water solution. This
co-solvent system dissolves hydrophobic compounds well, and itself
produces low toxicity upon systemic administration. Naturally, the
proportions of a co-solvent system may be varied considerably
without destroying its solubility and toxicity characteristics.
Furthermore, the identity of the co-solvent components may be
varied: for example, other low-toxicity nonpolar surfactants may be
used instead of polysorbate 80; the fraction size of polyethylene
glycol may be varied; other biocompatible polymers may replace
polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars
or polysaccharides may be substituted for dextrose.
[0134] Alternatively, other delivery systems for hydrophobic
pharmaceutical compounds may be employed. Liposomes and emulsions
are known examples of delivery vehicles or carriers for hydrophobic
drugs. Certain organic solvents such as dimethylsulfoxide also may
be employed, although usually at the cost of greater toxicity.
Additionally, the compounds may be delivered using a
sustained-release system, such as semipermeable matrices of solid
hydrophobic polymers containing the therapeutic agent. Various
sustained-release materials have been established and are known by
those skilled in the art. Sustained-release capsules may, depending
on their chemical nature, release the compounds for a few weeks up
to over 100 days. Depending on the chemical nature and the
biological stability of the therapeutic reagent, additional
strategies for protein stabilization may be employed.
[0135] The pharmaceutical compositions also may comprise suitable
solid- or gel-phase carriers or excipients. Examples of such
carriers or excipients include calcium carbonate, calcium
phosphate, sugars, starches, cellulose derivatives, gelatin, and
polymers such as polyethylene glycols.
[0136] Some of the compounds of the invention may be provided as
salts with pharmaceutically compatible counter ions.
Pharmaceutically compatible salts may be formed with many acids,
including hydrochloric, sulfuric, acetic, lactic, tartaric, malic,
succinic, etc. Salts tend to be more soluble in aqueous or other
protonic solvents than are the corresponding free-base forms.
[0137] The preparation of preferred compounds of the present
invention is described in detail in the following examples, but the
artisan will recognize that the chemical reactions described may be
readily adapted to prepare a number of other protein kinase
inhibitors of the invention. For example, the synthesis of
non-exemplified compounds according to the invention may be
successfully performed by modifications apparent to those skilled
in the art, e.g., by appropriately protecting interfering groups,
by changing to other suitable reagents known in the art, or by
making routine modifications of reaction conditions. Alternatively,
other reactions disclosed herein or known in the art will be
recognized as having applicability for preparing other compounds of
the invention.
EXAMPLES
[0138] In the examples described below, unless otherwise indicated
all temperatures are set forth in degrees Celsius and all parts and
percentages are by weight. Reagents were purchased from commercial
suppliers such as Aldrich Chemical Company or Lancaster Synthesis
Ltd. and were used without further purification unless otherwise
indicated. Tetrahydrofuran (THF) and N,N-dimethylforamide (DMF)
were purchased from Aldrich in Sure seal bottles and used as
received. All solvents were purified using standard methods readily
known to those skilled in the art, unless otherwise indicated.
[0139] The reactions set forth below were done generally under a
positive pressure of argon or nitrogen or with a drying tube, at
ambient temperature (unless otherwise stated), in anhydrous
solvents, and the reaction flasks were fitted with rubber septa for
the introduction of substrates and reagents via syringe. Glassware
was oven dried and/or heat dried. Analytical thin layer
chromatography (TLC) was performed on glass-backed silica gel 60 F
254 plates and eluted with the appropriate solvent ratios (v/v),
and are denoted where appropriate. The reactions were assayed by
TLC and terminated as judged by the consumption of starting
material.
[0140] Visualization of the TLC plates was generally done by
ultraviolet visualization. Work-ups were typically done by doubling
the reaction volume with the reaction solvent or extraction solvent
and then washing with the indicated aqueous solutions using 25% by
volume of the extraction volume unless otherwise indicated. Product
solutions were dried over anhydrous Na.sub.2SO.sub.4 prior to
filtration and evaporation of the solvents under reduced pressure
on a rotary evaporator and noted as solvents removed in vacuo.
Products were purified by employing radial chromatography or flash
column chromatography (Still et al., J. Org. Chem., 43, 2923
(1978)), the latter using Merck grade flash silica gel (47-61
.mu.m) and a silica gel: crude material ratio of about 20:1 to
100:1 unless otherwise stated. Hydrogenolysis was done at the
pressure indicated in the examples or at ambient pressure.
[0141] .sup.1H-NMR spectra were recorded on an instrument operating
at 300 or 500 MHz, and .sup.13C-NMR spectra were recorded operating
at 75 MHz. NMR spectra were obtained as CDCl.sub.3 solutions
(reported in ppm), using chloroform as the reference standard (7.25
ppm and 77.00 ppm) or CD.sub.3OD (3.4 and 4.8 ppm and 49.3 ppm), or
internally tetramethylsilane (0.00 ppm) when appropriate. Other NMR
solvents were used as needed. When peak multiplicities are
reported, the following abbreviations are used: s (singlet), d
(doublet), t (triplet), m (multiplet), q (quartet), br (broadened),
dd (doublet of doublets), dt (doublet of triplets). Coupling
constants, when given, are reported in Hertz (Hz).
[0142] Infrared (IR) spectra were recorded on a Perkin-Elmer FT-IR
Spectrometer as neat oils, as KBr pellets, or as CDCl.sub.3
solutions, and when given are reported in wave numbers (cm.sup.-1).
All melting points (mp) are uncorrected.
[0143] Unless otherwise stated, the HPLC conditions are the
following: Hewlett Packard ODS Hypersil (5 .mu.m, 125.times.4 mm),
10% acetonitrile/0.1 M ammonium acetate from 0-2 minutes to 90%
acetonitrile/0.1 M ammonium acetate at 22 minutes, 1.0 mL/minute,
detection at 254 nm.
[0144] Abbreviations for reagents, equipment, and techniques are
defined as follows: MTBE (methyl t-butyl ether); DMSO
(dimethylsulfoxide); DIEA (diisopropylethylamine); TEA
(triethylamine); AcOH (acetic acid); DMAP
(4-(dimethylamino)pyridine); EDC
(1-ethyl-3-(3'-dimethylaminopropyl)carbo- diimide HCl); HATU
(O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluroniu- m
hexafluorophosphate); HOBt (N-hydroxybenzotriazole); PyBop
(benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium
hexafluorophosphate); MS (ESI) (Electrospray ionization mass
spectrometry); MS (FAB) (fast atom bombardment mass spectrometry);
HRMS (FAB) (high resolution fast atom bombardment mass
spectrometry); HRMS (MALDI) (high resolution matrix-assisted laser
desorption/ionization mass spectrometry); and APCIMS (atmospheric
pressure chemical ionization mass spectrometry).
EXAMPLE A-1
N-(3,4,5-Trimethoxyphenyl)-3-[(pyrazin-2-yl)sulfanylmethyl]benzamide
[0145] 20
[0146] (a) To a solution of 3,4,5-trimethoxyaniline (3.0 g, 16.4
mmol) and triethylamine (2.5 mL, 18 mmol) in 100 mL dichloromethane
at room temperature was added, via pipette,
3-(chloro-methyl)benzoyl chloride (2.4 mL, 16.9 mmol). After
stirring at room temperature for 4 h, the mixture was partitioned
between chloroform (100 mL) and water (200 mL). The aqueous layer
was extracted twice with chloroform (100 mL) and the combined
organics were washed with brine (100 mL), dried over
Na.sub.2SO.sub.4, and concentrated to dryness. The crude residue
was triturated with MTBE to obtain 5.22 g (95%) of
N-(3,4,5-trimethoxyphenyl)- -3-(chloromethyl)benzamide, A-1a, as an
off-white solid which was collected by filtration: mp
138-145.degree. C.; .sup.1H NMR (DMSO-d.sub.6) .delta. 7.98 (s,
1H), 7.89 (d, 1H, J=7.8 Hz), 7.63 (d, 1H, J=7.8 Hz), 7.52 (t, 1H,
J=7.7 Hz), 7.20 (s, 2H), 4.83 (s, 2H), 3.75 (s, 6H), 3.62 (s, 3H).
Anal. calc'd for C.sub.17H.sub.18NO.sub.4Cl.0.2H.sub.2- O: C,
60.16; H, 5.47; N, 4.13; Cl, 10.45. Found: C, 60.18; H, 5.38; N,
4.17; Cl, 10.68.
[0147] (b) To a solution of 0.112 g (1 mmol) of 2-pyrazinethiol
(Specs) and N-(3,4,5-trimethoxyphenyl)-3-(chloromethyl)benzamide,
A-1a, (0.335 g, 1 mmol) in 5 mL anhydrous DMF under an argon purge
was added cesium carbonate (0.814 g, 2.5 mmol). The resulting
suspension was stirred at .about.65.degree. C. for 17 hr. The
mixture was allowed to cool to room temperature and then was
partitioned between ethyl acetate (50 mL) and water (75 mL). The
aqueous layer was extracted twice with ethyl acetate (50 mL) and
the combined organics were washed with brine (25 mL), dried over
Na.sub.2SO.sub.4, and concentrated to dryness. The crude residue
was purified on silica gel using a gradient of 0% to 6% methanol in
1:1 ethyl acetate:hexane as eluent to obtain
N-(3,4,5-trimethoxyphenyl)-3-[(pyrazin-
-2-yl)sulfanyl-methyl]benzamide as a pale yellow oil (0.18 g, 43%)
which crystallized upon standing: mp 112-119.degree. C.; .sup.1H
NMR (DMSO-d.sub.6) .delta. 10.13 (s, 1H), 8.63 (d, 1H, J=1.55 Hz),
8.53 (dd, 1H, J=2.60, 1.58 Hz), 8.36 (d, 1H, J=2.64 Hz), 8.00 (s,
1H), 7.83 (d, 1H, J=7.81 Hz), 7.64 (d, 1H, J=7.72 Hz), 7.47 (t, 1H,
J=7.7 Hz), 7.21 (s, 2H), 4.56 (s, 2H), 3.77 (s, 6H), 3.65 (s, 3H).
Anal. calc'd for C.sub.21H.sub.21N.sub.3O.sub.4S.0.2 MTBE: C,
61.58; H, 5.50; N, 9.79; S, 7.47. Found: C, 61.34; H, 5.43; N,
9.69; S, 7.34.
EXAMPLE A-2
N-(3,4,5-Trimethoxyphenyl)-3-[(5-amino-2H-[1,2,4]triazol-3-yl)
sulfanylmethyl]benzamide
[0148] 21
[0149] Example A-2 was prepared in a similar manner to that
described for A-1, except that 3-amino-5-mercapto-1,2,4-triazole
(Aldrich) was used in place of 2-pyrazinethiol in step (b): .sup.1H
NMR (DMSO-d.sub.6) .delta. 11.96 (br s, 1H), 10.12 (s, 1H), 7.95
(s, 1H), 7.82 (d, 1H, J=7.8 Hz), 7.59 (d, 1H, J=7.6 Hz), 7.46 (t,
1H, J=7.69 Hz), 7.23 (s, 2H), 6.05 (br s, 2H), 4.32 (s, 2H), 3.78
(s, 6H), 3.65 (s, 3H); HR MS (FAB): Calculated for
C.sub.19H.sub.22N.sub.5O.sub.4S (M+H.sup.+): 416.1393. Found:
416.1408. Anal. calc'd for C.sub.19H.sub.21N.sub.5O.sub.4S.0.3
EtOAc: C, 54.90; H, 5.34; N, 15.85; S, 7.26. Found: C, 54.87; H,
5.50; N, 15.71; S, 7.03.
EXAMPLE A-3
N-(4-Isopropyl-3-methylphenyl)-3-[(pyrazin-2-yl)sulfanylmethyl]-Benzamide
[0150] 22
[0151] Example A-3 was prepared in a similar manner to that
described for A-1, except that 3-methyl-4-isopropylaniline
hydrochloride (Maybridge) was used in place of
3,4,5-trimethoxyaniline in step (a): mp 69-73.degree. C.; .sup.1H
NMR (DMSO-d.sub.6) .delta. 10.07 (s, 1H), 8.62 (d, 1H, J=1.6 Hz),
8.52 (dd, 1H, J=2.6, 1.6 Hz), 8.35 (d, 1H, J=2.6 Hz), 7.99 (s, 1H),
7.83 (d, 1H, J=7.8 Hz), 7.62 (d, 1H, J=7.7 Hz), 7.56-7.51 (m, 2H),
7.46 (t, 1H, J=7.7 Hz), 7.20 (d, 1H, J=8.3 Hz), 4.55 (s, 2H),
3.10-3.05 (m, 1H), 2.69 (s, 3H), 1.17 (d, 6H, J=6.9 Hz). Anal.
calc'd for C.sub.22H.sub.23N.sub.3OS.0.2 MTBE: C, 69.91; H, 6.48;
N, 10.64; S, 8.12. Found: C, 70.03; H, 6.40; N, 10.41; S, 7.81.
EXAMPLE A-4
N-(4-Isopropyl-3-methylphenyl)-3-[(5-amino-2H-[1,2,4]triazol-3-yl)sulfanyl-
methyl]Benzamide
[0152] 23
[0153] Example A-4 was prepared in a similar manner to that
described for A-1, except that 3-methyl-4-isopropylaniline was used
in place of 3,4,5-trimethoxyaniline in step (a), and
3-amino-5-mercapto-1,2,4-triazol- e was used in place of
2-pyrazinethiol in step (b): .sup.1H NMR (DMSO-d.sub.6) .delta.
11.93 (br s, 1H), 10.05 (s, 1H), 7.92 (s, 1H), 7.80 (d, 1H, J=7.8
Hz), 7.57-7.51., (m, 3H), 7.43 (t, 1H, J=7.7 Hz), 7.19 (d, 1H,
J=8.3 Hz), 6.02 (br s, 2H), 4.30 (s, 2H), 3.09-3.04 (m, 1H), 2.29
(s, 3H), 1.17 (d, 6H, J=6.9 Hz). Anal. calc'd for
C.sub.20H.sub.23N.sub.5- OS.0.1 MTBE: C, 63.08; H, 6.25; N, 17.94;
S, 8.22. Found: C, 62.78; H, 6.26; N, 17.78; S, 8.00.
EXAMPLE A-5
N-(4-Isopropyl-3-methylphenyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfan-
ylmethyl]Benzamide
[0154] 24
[0155] Example A-5 was prepared in a similar manner to that
described for A-1, except that 3-methyl-4-isopropylaniline was used
in place of 3,4,5-trimethoxyaniline in step (a), and
4-mercapto-1H-pyrazolo[3,4-d]pyr- imidine was used in place of
2-pyrazinethiol in step (b): mp 187-189.degree. C.; .sup.1H NMR
(DMSO-d.sub.6) .delta. 10.09 (s, 1H), 8.81 (s, 1H), 8.31 (s, 1H),
8.05 (s, 1H), 7.85 (d, 1H, J=7.9 Hz), 7.69 (d, 1H, J=7.7 Hz),
7.55-7.47 (m, 3H), 7.20 (d, 1H, J=8.4 Hz), 4.78 (s, 2H), 3.09-3.06
(m, 1H), 2.29 (s, 3H), 1.18 (d, 6H, J=6.9 Hz). Anal. calc'd for
C.sub.23H.sub.23N.sub.5OS: C, 66.16; H, 5.55; N, 16.77; S, 7.68.
Found: C, 65.90; H, 5.51; N, 16.98; S, 7.40.
EXAMPLE A-6
N-(2-Methylquinolin-6-yl)-3-[(pyrazin-2-yl)sulfanylmethyl]Benzamide
[0156] 25
[0157] Example A-6 was prepared in a similar manner to that
described for A-1, except that 6-amino-2-methylquinoline
(Lancaster) was used in place of 3,4,5-trimethoxyaniline in step
(a): mp 133-135.degree. C.; .sup.1H NMR (DMSO-d.sub.6) .delta.
10.51 (s, 1H), 8.63 (s, 1H), 8.53 (t, 1H, J=1.9 Hz), 8.44 (t, 1H,
J=3.0 Hz), 8.35 (d, 1H, J=2.6 Hz), 8.20 (d, 1H, J=2.4 Hz), 8.04 (s,
1H), 7.96 (d, 1H, J=9.0 Hz), 7.89 (d, 2H, J=9.1 Hz), 7.66 (d, 1H,
J=7.7 Hz), 7.50 (t, 1H, J=7.7 Hz), 7.38 (d, 1H, J=8.4 Hz), 4.57 (s,
2H), 2.63 (s, 3H). Anal. calc'd for C.sub.22H.sub.18N.sub.4OS: C,
68.37; H, 4.69; N, 14.50; S, 8.30. Found: C, 68.41; H, 4.72; N,
14.52; S, 8.30.
EXAMPLE A-7
N-(3-Isopropylphenyl)-3-[(pyrazin-2-yl)sulfanylmethyl]Benzamide
[0158] 26
[0159] Example A-7 was prepared in a similar manner to that
described for A-1, except that 3-isopropylaniline (Maybridge) was
used in place of 3,4,5-trimethoxyaniline in step (a): .sup.1H NMR
(DMSO-d.sub.6) .delta. 10.02 (s, 1H), 8.63 (d, 1H, J=1.6 Hz), 8.52
(dd, 1H, J=2.42, 1.5 Hz), 8.34 (d, 1H, J=2.6 Hz), 7.99 (s, 1H),
7.83 (d, 1H, J=7.7 Hz), 7.63-7.60 (m, 3H), 7.46 (t, 1H, J=7.7 Hz),
7.25 (t, 1H, J=7.8 Hz), 6.97 (d, 1H, J=7.6 Hz), 4.55 (s, 2H),
2.88-2.85 (m, 1H), 1.21 (d, 6H, J=6.9 Hz). Anal. calc'd for
C.sub.21H.sub.21N.sub.3OS.0.3 MTBE: C, 69.31; H, 6.36; N, 10.78; S,
8.22. Found: C, 69.34; H, 6.15; N, 10.54; S, 7.96.
EXAMPLE A-8
N-(3,5-Dibromo-4-methylphenyl)-3-[(pyrazin-2-yl)sulfanyl-methyl]Benzamide
[0160] 27
[0161] Example A-8 was prepared in a similar manner to that
described for A-1, except that 3,5-dibomo-4-methylaniline
(Lancaster) was used in place of 3,4,5-trimethoxyaniline in step
(a): mp 119-127.degree. C.; .sup.1H NMR (DMSO-d.sub.6) .delta.
10.39 (s, 1H), 8.61 (d, 1H, J=1.30 Hz), 8.51 (t, 1H, J=2.1 Hz),
8.34 (d, 1H, J=2.61 Hz), 8.12 (s, 2H), 7.99 (s, 1H), 7.82 (d, 1H,
J=7.96 Hz), 7.65 (d, 1H, J=7.73 Hz), 7.48 (t, 1H, J=7.74 Hz), 4.54
(s, 2H), 2.47 (s, 3H). Anal. calc'd for C.sub.19H.sub.15N.sub.3-
4S.0.25 EtOAc: C, 46.62; H, 3.33; N, 8.16; S, 6.22. Found: C,
46.33; H, 3.24; N, 7.90; S, 5.83.
EXAMPLE B-1
N-(3,4,5-Trimethoxyphenyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylme-
thyl]Benzamide
[0162] 28
[0163] To a solution of 3,4,5-trimethoxyaniline (400 mg, 2.18 mmol)
and triethylamine (0.30 mL, 2.18 mmol) in dichloromethane was added
0.31 mL (2.18 mmol) of 3-chloromethylbenzoyl chloride (Aldrich).
After 10 min, the solvent was removed and the residual crude
N-(3,4,5-trimethoxyphenyl)- -3-(chloromethyl)benzamide, A-1a, was
dissolved in DMF (10 mL) under argon. To the resulting solution was
added 4-mercapto-1H-pyrazolo[3,4-d]p- yrimidine (332 mg, 2.18 mmol)
followed by triethylamine (0.30 mL, 2.18 mmol). The resulting
solution was heated at 70.degree. C. for 2 h, then cooled and
poured into water. The solid was collected by filtration and washed
with water. After air-drying, the solid was sequentially triturated
with ethyl acetate/hexane and with dichloromethane, and the solid
collected by filtration to provide 360 mg (37%) of
N-(3,4,5-trimethoxyphenyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylm-
ethyl)benzamide, B-1: .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
14.12 (s, 1H), 10.15 (s, 1H), 8.79 (s, 1H), 8.03 (s, 1H), 7.83 (d,
1H, J=7.7 Hz), 7.68 (d, 1H, J=7.7 Hz), 7.48 (t, 1H, J=7.7 Hz), 7.20
(s, 2H), 4.77 (s, 2H), 3.75 (s, 6H), 3.62 (s, 3H). Anal. calc'd for
C.sub.22H.sub.21N.sub.5O.sub.4S.0.7H.sub.2O: C, 56.93; H, 4.87; N,
15.09; S, 6.91. Found: C, 56.89; H, 4.76; N, 14.85; S, 6.91.
EXAMPLE B-2
N-(3,4,5-Trimethoxyphenyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylme-
thyl]Benzamide
[0164] 29
[0165] Example B-2 was prepared in a similar manner to that
described for B-1, except that 5-amino-2-methoxypyridine was used
in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.10 (s, 1H), 10.20 (s, 1H), 8.75 (s, 1H),
8.43 (d, J=2.5 Hz, 1H), 8.25 (s, 1H), 7.95-8.00 (m, 2H), 7.81 (d,
1H, J=7.8 Hz), 7.65 (d, 1H, J=7.7 Hz), 7.41-7.46. (dd, 1H), 6.78
(d, 1H, J=8.8 Hz), 4.70 (s, 2H), 3.80 (s, 3H); APCIMS m/z 393
[M+H].sup.+.
EXAMPLE B-3
N-(Quinolin-6-yl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanyl-methyl]Ben-
zamide
[0166] 30
[0167] Example B-3 was prepared in a similar manner to that
described for B-1, except that 6-aminoquinoline was used in place
of 3,4,5-trimethoxyaniline: mp 236-240.degree. C. (dec); .sup.1H
NMR (DMSO-d.sub.6) .delta. 14.15 (s, 1H), 10.60 (s, 1H), 8.80 (s,
2H), 8.52 (s, 1H), 8.34-8.29 (m, 2H), 8.10 (s, 1H), 8.05-7.98 (m,
2H), 7.91 (d, 1H, J=7.7 Hz), 7.72 (d, 1H, J=7.4 Hz), 7.54-7.48 (m,
2H), 4.79 (s, 2H). Anal. calc'd for
C.sub.22H.sub.16N.sub.6OS.0.7H.sub.2O: C, 62.16; H, 4.13; N, 19.77;
S, 7.54. Found: C, 62.34; H, 3.83; N, 19.48; S, 7.61.
EXAMPLE B-4
N-(5-Methylisoxazol-3-yl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmet-
hyl]Benzamide
[0168] 31
[0169] Example B-4 was prepared in a similar manner to that
described for B-1, except that 3-amino-5-methylisoxazole was used
in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 13.99 (s, 1H), 11.34 (s, 1H), 8.79 (s, 1H),
8.31 (s, 1H), 8.11 (s, 1H), 7.90 (d, 1H, J=8.0 Hz), 7.72 (d, 1H,
J=8.0 Hz), 7.47 (dd, 1H, J=7.5, 7.6 Hz), 6.74 (s, 1H), 4.76 (s,
2H), 2.41 (s, 3H); APCIMS m/z 367 [M+H].sup.+.
EXAMPLE B-5
N-(Pyridin-4-yl)methyl-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-sulfanylmethy-
l]Benzamide
[0170] 32
[0171] Example B-5 was prepared in a similar manner to that
described for B-1, except that 4-picolylamine was used in place of
3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 14.05 (s, 1H), 9.08-9.12 (t, 1H, J=5.8 Hz), 8.74 (s, 1H),
8.44 (d, 1H, J=5.7 Hz), 8.25 (s, 1H), 7.96 (s, 1H), 7.76 (d, 1H,
J=7.9 Hz), 7.62 (d, 1H, J=7.7 Hz), 7.40 (dd, 1H, J=7.7, 7.9 Hz),
7.24 (d, 1H, J=5.7 Hz), 4.70 (s, 2H), 4.43 (d, 2H, J=5.9 Hz);
APCIMS m/z 377 [M+H].sup.+.
EXAMPLE B-6
N-(1,3-Benzodioxyl-5-ylmethyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-sulfa-
nylmethyl]Benzamide
[0172] 33
[0173] Example B-6 was prepared in a similar manner to that
described for B-1, except that 3,4-(methylenedioxy)benzylamine was
used in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.05 (s, 1H), 8.98 (t, 1H, J=6.0 Hz), 8.78
(s, 1H), 8.30 (s, 1H), 7.99 (s, 1H), 7.77 (d, 1H, J=7.9 Hz), 7.64
(d, 1H, J=7.5 Hz), 7.42 (dd, 1H, J=7.6, 7.9 Hz), 6.84-6.87 (m, 2H),
6.78 (d, 1H, J=7.9 Hz), 5.97 (s, 2H), 4.74 (s, 2H), 4.36 (d, 2H,
J=6.1 Hz); APCIMS m/z 420 [M+H].sup.+.
EXAMPLE B-7
N-(2-Methoxybenzyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-sulfanylmethyl]B-
enzamide
[0174] 34
[0175] Example B-7 was prepared in a similar manner to that
described for B-1, except that 2-methoxybenzylamine was used in
place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.12 (s, 1H), 8.86 (t, 1H, J=6.0 Hz), 8.79
(s, 1H), 8.30 (s, 1H), 8.02 (s, 1H), 7.81 (d, 1H, J=7.9 Hz), 7.64
(d, 1H, J=7.6 Hz), 7.44 (dd, 1H, J=7.6, 7.9 Hz), 7.23 (m, 1H), 7.16
(d, 1H, J=7.2 Hz), 6.98 (d, 1H, J=7.9 Hz), 6.89 (dd, 1H, J=7.5, 6.8
Hz), 4.75 (s, 2H), 4.43 (d, 2H, J=6.1 Hz), 3.82 (s, 3H); APCIMS m/z
406 [M+H].sup.+.
EXAMPLE B-8
N-(2-Phenylethyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]Benz-
amide
[0176] 35
[0177] Example B-8 was prepared in a similar manner to that
described for B-1, except that phenethylamine was used in place of
3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 14.13 (s, 1H), 8.79 (s, 1H), 8.61 (t, 1H J=5.7 Hz), 8.30
(s, 1H), 7.93 (s, 1H), 7.70 (d, 1H, J=7.9 Hz), 7.62 (d, 1H, J=7.9
Hz), 7.41 (dd, 1H, J=7.6, 7.9 Hz), 7.16-7.31 (m, 5H), 4.73 (s, 2H),
3.43-3.50 (m, 2H), 2.83 (dd, 2H, J=7.2, 7.9 Hz); APCIMS m/z 390
[M+H].sup.+.
EXAMPLE B-9
N-(2-Methoxyphenyl)-3-[(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-sulfanylmethyl]B-
enzamide
[0178] 36
[0179] Example B-9 was prepared in a similar manner to that
described for B-1, except that 2-methoxyaniline was used in place
of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 14.12 (s, 1H), 9.43 (s, 1H), 8.81 (s, 1H), 8.32 (s, 1H),
8.08 (s, 1H), 7.87 (d, 1H, J=7.9 Hz), 7.77 (d, 1H, J=7.5 Hz), 7.70
(d, 1H, J=7.9 Hz), 7.48 (dd, 1H, J=7.5, 7.6 Hz), 7.19 (m, 1H), 7.09
(d, 1H, J=7.2 Hz), 6.97 (dd, 1H, J=7.1, 8.0 Hz), 4.79 (s, 2H), 3.83
(s, 3H); APCIMS in/z 392 [M+H].sup.+.
EXAMPLE B-10
N-[3-(N-Methyl-N-phenylamino)propyl]-3-[(5-methyl-1H-1,2,4-triazol-3-yl)su-
lfanylmethyl]Benzamide
[0180] 37
[0181] Example B-10 was prepared in a similar manner to that
described for B-1, except that N-(3-aminopropyl)-N-methylaniline
was used in place of 3,4,5-trimethoxyaniline, and
3-mercapto-5-methyl-1H-1,2,4-triazole was used in place of
4-mercapto-1H-pyrazolo[3,4-d]pyrimidine: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.80 (s, 1H), 7.68 (d, 1H, J=7.9 Hz), 7.52 (d,
1H, J=7.5 Hz), 7.38 (dd, 1H, J=7.5, 8.0 Hz), 7.15 (m, 2H),
6.74-6.77 (m, 2H), 6.64 (dd, 1H, J=7.1, 7.1 Hz).4.36 (s, 2H),
3.41-3.45 (m, 4H), 2.93 (s, 3H), 2.38 (s, 3H), 1.85-1.96 (m, 2H);
APCIMS m/z 396 [M+H].sup.+.
EXAMPLE B-11
N-(1,3-Benzodioxyl-5-ylmethyl)-3-[(5-methyl-1H-1,2,4-triazol-3-yl)sulfanyl-
methyl]Benzamide
[0182] 38
[0183] Example B-11 was prepared in a similar manner to that
described for B-1, except that (3,4-methylenedioxy)benzylamine was
used in place of 3,4,5-trimethoxyaniline, and
3-mercapto-5-methyl-1H-1,2,4-triazole was used in place of
4-mercapto-1H-pyrazolo[3,4-d]pyrimidine: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.83 (s, 1H), 7.71 (d, 1H, J=7.5 Hz), 7.52 (d,
1H, J=7.2 Hz), 7.38 (dd, 1H, J=7.50, 7.6 Hz), 6.76-6.86 (m, 3H),
5.93 (s, 2H), 4.47 (s, 2H), 4.35 (s, 2H), 2.38 (s, 3H); APCIMS m/z
383 [M+H].sup.+.
EXAMPLE B-12
N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-y-
l-sulfanyl)methyl]Benzamide
[0184] 39
[0185] Example B-12 was prepared in a similar manner to that
described for B-1, except that 4-cyano-3-trifluoromethylaniline was
used in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.07 (s, 1H), 10.98 (s, 1H), 8.80 (s, 1H),
8.44 (s, 1H), 8.26-8.31 (m, 2H), 8.15 (d, 1H, J=7.9 Hz), 8.08 (s,
1H), 7.89 (d, 1H, J=7.5 Hz), 7.76 (d, 1H, J=7.9 Hz), 7.53 (dd, 1H
J=7.50, 7.9 Hz), 4.79 (s, 2H); APCIMS m/z 455 [M+H].sup.+.
EXAMPLE B-13
N-(3,3-Diphenylpropyl)-3-{[(5-methyl-1H-1,2,4-triazol-3-yl)-sulfanyl]Methy-
l}Benzamide
[0186] 40
[0187] Example B-13 was prepared in a similar manner to that
described for B-1, except that 3,3-diphenyl-1-propylamine was used
in place of 3,4,5-trimethoxyaniline, and
3-mercapto-5-methyl-1H-1,2,4-triazole was used in place of
4-mercapto-1H-pyrazolo[3,4-d]pyrimidine: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.74 (s, 1H), 7.62 (d, 1H, J=7.6 Hz), 7.50 (d,
1H, J=7.6 Hz), 7.25-7.38 (m, 9H), 7.13-7.18 (m, 2H), 4.35 (s, 2H),
4.05 (dd, 1H, J=7.6, 7.9 Hz), 3.34-3.37 (m, 2H), 2.38 (m, 5H);
APCIMS m/z 443 [M+H].sup.+.
EXAMPLE B-14
3-{[(5-Methyl-1H-1,2,4-triazol-3-yl)-sulfonyl]methyl}-N-phenethylbenzamide
[0188] 41
[0189] Example B-14 was prepared in a similar manner to that
described for B-1, except that 2-phenylethylamine was used in place
of 3,4,5-trimethoxyaniline, and
3-mercapto-5-methyl-1H-1,2,4-triazole was used in place of
4-mercapto-1H-pyrazolo[3,4-d]pyrimidine: .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.78 (s, 1H), 7.64 (d, 1H, J=7.9 Hz), 7.51 (d,
1H, J=7.5 Hz), 7.38 (dd, 1H, J=7.5, 7.9 Hz), 7.18-7.33 (m, 5H),
4.35 (s, 2H), 3.57-3.62 (m, 2H), 2.90-2.93 (m, 2H), 2.40 (s, 3H);
APCIMS m/z 353 [M+H].sup.+.
EXAMPLE B-15
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-isopropylphenyl)--
benzamide
[0190] 42
[0191] Example B-15 was prepared in a similar manner to that
described for B-1, except that 3-isopropylaniline was used in place
of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 14.13 (s, 1H), 10.19 (s, 1H), 8.80 (s, 1H), 8.31 (s, 1H),
8.05 (s, 1H), 7.85 (d, 1H, J=7.9 Hz), 7.69 (d, 1H, J=7.9 Hz), 7.61
(m, 2H), 7.48 (dd, 1H, J=7.6, 7.6 Hz), 7.25 (dd, 1H, J=7.5, 8.0
Hz), 6.99 (d, 1H, J=7.9 Hz), 4.78 (s, 2H), 2.83-2.91 (m, 1H), 1.21
(d, 6H, J=6.0 Hz); APCIMS m/z 404 [M+H].sup.+.
EXAMPLE B-16
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-trifluoromethyl-5-
-methoxyphenyl)-benzamide
[0192] 43
[0193] Example B-16 was prepared in a similar manner to that
described for B-1, except that
3-trifluoromethyl-5-trifluoromethoxyaniline was used in place of
3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 14.13 (s, 1H), 10.51 (s, 1H), 8.80 (s, 1H), 8.30 (s, 1H),
8.06 (s, 1H), 7.86 (d, 1H, J=7.9 Hz), 7.81 (s, 1H), 7.73 (m, 2H),
7.50 (dd, 1H, J=7.50, 8.0 Hz), 6.98 (s, 2H), 6.46-6.50 (m, 1H),
4.78 (s, 2H), 3.83 (s, 3H); APCIMS m/z 460 [M+H].sup.+.
EXAMPLE B-17
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3,5-bis-trifluorome-
thylphenyl)-benzamide
[0194] 44
[0195] Example B-17 was prepared in a similar manner to that
described for B-1, except that 3,5-bis(trifluoromethyl)aniline was
used in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.20 (s, 1H), 10.92 (s, 1H), 8.87 (s, 1H),
8.57 (s, 2H), 8.37 (s, 1H), 8.17 (s, 1H), 7.97 (d, 1H, J=7.9 Hz),
7.89 (s, 1H), 7.83 (d, 1H, J=7.6 Hz), 7.60 (dd, 1H, J=7.50, 8.0
Hz), 4.86 (s, 2H); APCIMS m/z 498 [M+H].sup.+.
EXAMPLE B-18
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-t-butylphenyl)-be-
nzamide
[0196] 45
[0197] Example B-18 was prepared in a similar manner to that
described for B-1, except that 3-(tert-butyl)aniline was used in
place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.13 (s, 1H), 10.19 (s, 1H), 8.81 (s, 1H),
8.30 (s, 2H), 8.05 (s, 1H), 7.85-7.88 (d, 1H, J=7.6 Hz), 7.75 (s,
1H), 7.65-7.71 (m, 1H), 7.48 (dd, 1H, J=7.6, 7.9 Hz), 7.26 (dd, 1H,
J=7.5, 8.0 Hz), 7.13 (d, 1H, J=7.9 Hz), 4.78 (s, 2H), 1.29 (s, 9H);
APCIMS t/z 418 [M+H].sup.+.
EXAMPLE B-19
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(4-isopropylphenyl)--
benzamide
[0198] 46
[0199] Example B-19 was prepared in a similar manner to that
described for B-1, except that 4-isopropylaniline was used in place
of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 14.13 (s, 1H), 10.19 (s, 1H), 8.80 (s, 1H), 8.30 (s, 1H),
8.04 (s, 1H), 7.84 (d, 1H, J=7.6 Hz), 7.64-7.70 (m, 3H), 7.47 (dd,
1H, J=7.5, 8.0 Hz), 7.21 (d, 2H, J=8.3 Hz), 4.78 (s, 2H), 2.81-2.90
(m, 1H), 1.19 (d, 6H, J=6.0 Hz); APCIMS m/z 404 [M+H].sup.+.
EXAMPLE B-20
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(4-trifluoromethoxyp-
henyl)-benzamide
[0200] 47
[0201] Example B-20 was prepared in a similar manner to that
described for B-1, except that 3-trifluoromethoxyaniline was used
in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.13 (s, 1H), 10.54 (s, 1H), 8.80 (s, 1H),
8.30 (s, 1H), 8.06 (s, 1H), 7.93 (s, 1H), 7.86 (d, 1H, J=7.6 Hz),
7.71-7.78 (m, 2H), 7.46-7.53 (m, 2H), 7.09 (d, 1H, J=8.3 Hz), 4.79
(s, 2H); APCIMS m/z 446 [M+H].sup.+.
EXAMPLE B-21
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3,5-dimethylphenyl)-
-benzamide
[0202] 48
[0203] Example B-21 was prepared in a similar manner to that
described for B-1, except that 3,5-dimethylaniline was used in
place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.13 (s, 1H), 10.10 (s, 1H), 8.80 (s, 1H),
8.30 (s, 1H), 8.04 (s, 1H), 7.83-7.85 (d, 1H, J=7.5 Hz), 7.68-7.71
(d, 1H, J=7.6 Hz), 7.45-7.50 (dd, 1H, J=7.50, 7.6 Hz), 7.39 (s,
2H), 6.74 (s, 1H), 4.78 (s, 2H), 2.26 (s, 6H); APCIMS m/z 390
[M+H].sup.+.
EXAMPLE B-22
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-(2-hydroxyethyl)p-
henyl)-benzamide
[0204] 49
[0205] Example B-22 was prepared in a similar manner to that
described for B-1, except that 3-(1-hydroxyethyl)aniline was used
in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.13 (s, 1H), 10.23 (s, 1H), 8.80 (s, 1H),
8.30 (s, 1H), 8.06 (m, 1H), 7.86 (d, 1H, J=7.9 Hz), 7.73 (s, 1H),
7.74-7.84 (m, 2H), 7.48 (dd, 1H, J=7.6, 7.9 Hz), 7.27 (dd, 1H,
J=7.9, 8.0 Hz), 7.06 (d, 1H, J=7.9 Hz), 5.18 (d, 1H, J=3.0), 4.78
(s, 2H), 4.68 (q, 1H), 1.32 (d, 3H, J=9.0 Hz); APCIMS n/z 406
[M+H].sup.+.
EXAMPLE B-23
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(4-dimethylaminophen-
yl)-benzamide
[0206] 50
[0207] Example B-23 was prepared in a similar manner to that
described for B-1, except that 4-dimethylaminoaniline was used in
place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) 13.76 (s, 1H), 9.63 (s, 1H), 8.44 (s, 1H), 7.94 (s,
1H), 7.66 (s, 1H), 7.46 (d, 1H, J=7.9 Hz), 7.301 (d, 1H, J=7.5 Hz),
7.18 (d, 1H, J=9.0 Hz), 7.10 (dd, 1H, J=7.5, 7.6 Hz), 6.35 (d, 1H,
J=9.0 Hz), 4.56 (s, 2H), 2.13 (s, 6H); APCIMS m/z 405
[M+H].sup.+.
EXAMPLE B-24
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-trifluoromethylsu-
lfonylphenyl)-benzamide
[0208] 51
[0209] Example B-24 was prepared in a similar manner to that
described for B-1, except that 3-(trifluoromethylsulfonyl)aniline
was used in place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.13 (s, 1H), 10.83 (s, 1H), 8.80 (s, 1H),
8.66 (s, 1H), 8.35-8.39 (m, 1H), 8.30 (s, 1H), 8.09 (s, 1H), 7.90
(d, 1H, J=7.9 Hz), 7.85 (m, 2H), 7.75 (d, 1H, J=7.60), 7.53 (dd,
1H, J=7.5, 7.6 Hz), 4.79 (s, 2H); APCIMS m/z 494 [M+H].sup.+.
EXAMPLE B-25
3-[(1H-Pyrazolo[3,4-d]pyrimidin-4-yl)sulfanylmethyl]-N(3-dimethylaminophen-
yl)-benzamide
[0210] 52
[0211] Example B-25 was prepared in a similar manner to that
described for B-1, except that 3-dimethylaminoaniline was used in
place of 3,4,5-trimethoxyaniline: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) 14.14 (s, 1H), 10.06 (s, 1H), 8.80 (s, 1H), 8.30 (s,
1H), 8.04 (s, 1H), 7.84 (d, 1H, J=8.0 Hz), 7.680 (d, 1H, J=7.5 Hz),
7.46 (dd, 1H, J=7.5, 7.6 Hz), 7.20 (s, 1H), 7.13 (m, 2H), 6.46-6.50
(m, 1H), 4.78 (s, 2H), 2.89 (s, 6H); APCIMS m/z 405
[M+H].sup.+.
EXAMPLE B-26
[0212] (a) To an array of 40 .mu.L of 0.25 M solution of different
amines (0.01 mmol) in acetonitrile distributed in the eleven
columns of a 96-well plate was added 40 .mu.L of 0.25 M solution of
triethylamine (0.01 mmol) and the array of reactions was agitated
briefly. To each of the wells was added 40 .mu.L of a 0.25 M
solution of 3-(chloromethyl) benzoylchloride (0.01 mmol) in
acetonitrile and the plate was agitated in a shaker at room
temperature for 2 h.
[0213] (b) An 0.25 M solution of different mercapto compounds was
prepared in DMF and 40 .mu.L (0.01 mmol) and was added in eight
different rows to the appropriate intermediate from step (a) above.
To each reaction mixture was add approximately 8-15 mg of cesium
carbonate and the reactions were heated at 60.degree. C. on a
Vortex heater for 16 h. The solvents were removed using the
SpeedVac.TM. apparatus and the crude reaction mixtures were
redissolved in DMSO and transferred using a liquid handler to a 1
mL 96-well plate to give a final theoretical concentration of 10
mM.
EXAMPLE B-27
[0214] Using the general procedure described above in Example B-26,
the following compounds were made (wherein for convenience, and as
understood in the art, not all hydrogen atoms have been expressly
indicated as bonding to each carbon and/or nitrogen atom).
53545556575859606162636465-
66676869707172737475767778798081828384858687888990919293949596979899100101-
10210310410510610710810911011111211311411511611711811912012112212312412512-
6127128129130131132133
EXAMPLE C-1
3-[(5-Cyanoamino-2H-[1,2,4]triazol-3-yl)sulfanylmethyl]-N-(3,4,5-trimethox-
yphenyl)Benzamide
[0215] 134
[0216] To a suspension of 0.300 g (0.73 mmol) of
N-(3,4,5-trimethoxyphenyl-
)-3-[(5-amino-2H-[1,2,4]triazol-3-yl)sulfanylmethyl]benzamide, A-2,
in 10 mL THF was added ethanol until the mixture became
homogeneous. The mixture was cooled to 0.degree. C. and
4-methylmorpholine (0.095 ml, 0.86 mmol) was added followed by
cyanogen bromide (0.115 g, 1.08 mmol) in one portion. After
stirring at 0 to 20.degree. C. over 2 h, the mixture was
partitioned between ethyl acetate (50 mL) and brine (50 mL). The
aqueous layer was extracted twice with ethyl acetate (50 mL) and
the combined organics were washed with brine (25 mL), dried over
Na.sub.2SO.sub.4, and concentrated to dryness. The crude residue
was triturated with a mixture of MTBE, ethyl acetate, and hexanes
to yield the desired product as a pale yellow solid (0.27 g, 85%)
that was collected by filtration: mp>165.degree. C. (dec);
.sup.1H NMR (DMSO-d.sub.6) .delta. 10.14 (s, 1H), 8.19 (s, 1H),
7.96 (s, 1H), 7.84 (d, 1H, J=7.8 Hz), 7.62 (d, 1H, J=7.7 Hz), 7.49
(t, 1H, J=7.7 Hz), 7.18 (s, 2H), 4.39 (s, 2H), 3.77 (s, 6H), 3.69
(s, 3H). Anal. calc'd for C.sub.20H.sub.20N.sub.6O.sub.4S.0.3
EtOAc: C, 54.53; H, 4.84; N, 18.00; S, 6.87. Found: C, 54.86; H,
4.83; N, 17.91; S, 6.64.
EXAMPLE C-2
3-[(5-(Methoxycarbonylamino)-2H-[1,2,4]triazol-3-yl)sulfanylmethyl]-N-(3,4-
,5-Trimethoxyphenyl)Benzamide
[0217] 135
[0218] To a suspension of 0.154 g (0.37 mmol) of
N-(3,4,5-trimethoxyphenyl-
)-3-[(5-amino-2H-[1,2,4]triazol-3-yl)sulfanylmethyl]benzamide, A-2,
in dichloromethane (5 mL) at room temperature was added DMF until
the mixture became homogeneous. To the mixture was added
N-methylmorpholine (0.075 mL, 0.7 mmol) followed by methyl
chloroformate (0.050 mL, 0.65 mmol). After stirring room
temperature for 2 h, the mixture was partitioned between MTBE (50
mL) and brine (50 mL). The aqueous layer was extracted with 1:1
MTBE/ethyl acetate (2.times.50 mL) and the combined organics were
washed with brine (25 mL), dried over Na.sub.2SO.sub.4, and
concentrated to dryness. The crude residue was triturated with MTBE
and filtered. The solid was triturated a second time with
MTBE/ethyl acetate to yield
N-(3,4,5-trimethoxyphenyl)-3-[(5-methylcarbamoyl-2H-[1,2,4]triaz-
ol-3-yl) sulfanylmethyl]benzamide, C-2, as a white solid (0.13 g,
74%): mp>150.degree. C. (dec); .sup.1H NMR (DMSO-d.sub.6)
.delta. 10.10 (s, 1H), 7.97 (s, 1H), 7.84 (d, 1H, J=7.7 Hz), 7.61
(d, 1H, J=7.7 Hz), 7.48-7.45 (m, 2H), 7.21 (s, 2H), 4.38 (s, 2H),
3.90 (s, 3H), 3.77 (s, 6H), 3.64 (s, 3H). Anal. calc'd for
C.sub.21H.sub.23N.sub.5O.sub.6S.0.75H- .sub.2O: C, 51.79; H, 5.07;
N, 14.38; S, 6.58. Found: C, 52.13; H, 5.29; N, 14.11; S, 6.17.
EXAMPLE C-3
N-(3,4,5-Trimethoxyphenyl)-3-[(5-acetylamino-2H-[1,2,4]triazol-3-yl)sulfan-
ylmethyl]Benzamide
[0219] 136
[0220] To a solution of 0.15 g (0.37 mmol) of
N-(3,4,5-trimethoxyphenyl)-3-
-[(5-amino-2H-[1,2,4]triazol-3-yl)sulfanylmethyl]benzamide, A-2, in
acetic acid (5 mL) at room temperature was added acetic anhydride
(0.200 mL, 2.1 mmol). After stirring at room temperature for 1.5
hr, the mixture was added dropwise to a cold solution of phosphate
buffer (1M, pH 7, 60 mL). The resulting precipitate was collected
by filtration, washed with water, and dried under vacuum. The dried
solid was triturated with MTBE/ethyl acetate and filtered to yield
N-(3,4,5-trimethoxyphenyl)-3-[(5-acetylamin-
o-2H-[1,2,4]triazol-3-yl) sulfanylmethyl]benzamide, C-3, as a white
solid (0.12 g, 71%): mp 196-201.degree. C.; .sup.1H NMR
(DMSO-d.sub.6) .delta. 10.14 (s, 1H), 8.02 (s, 1H), 7.85 (d, 1H,
J=7.8 Hz), 7.65 (d, 1H, J=7.8 Hz), 7.49 (t, 1H, J=7.7 Hz), 7.22 (s,
2H), 4.40 (s, 2H), 3.78 (s, 6H), 3.65 (s, 3H), 2.50 (s, 3H,
obscured by DMSO). Anal. calc'd for
C.sub.21H.sub.23N.sub.5O.sub.5S.0.9H.sub.2O: C, 53.24; H, 5.28; N,
14.78; S, 6.77. Found: C, 53.28; H, 4.98; N, 14.48; S, 6.68.
EXAMPLE D-1
N-(4-Isopropyl-3-methylphenyl)-3-[(pyrazin-2-yl)methyl-sulfanyl]Benzamide
[0221] 137
[0222] (a) A suspension of methylpyrazine (5 g, 53 mmol) and
N-bromosuccinimide (9.45 g, 53 mmol) in carbon tetrachloride (200
mL) was heated at reflux while exposed to a 100 watt light source.
After 4 hr, the dark mixture was allowed to cool to room
temperature and was decanted. The supernatant was filtered and the
filtrate reduced to .about.25 mL volume then passed through silica
gel using a gradient of 0% to 5% ethyl acetate in CHCl.sub.3.
Decomposition was evident upon concentration of chromatographed
product. The residue was taken up in CH.sub.2Cl.sub.2, washed with
water, dried over sodium sulfate, and concentrated cold. Unstable
oily 2-(bromomethyl)pyrazine, D-1a, (2 g, 22%) was used quickly in
next reaction: .sup.1H NMR (CDCl.sub.3) .delta. 8.72 (s, 1H), 8.55
(d, 1H, J=1.8 Hz), 8.51 (d, 1H, J=2.5 Hz), 4.56 (s, 2H).
[0223] (b) 3-[(Pyrazin-2-yl)methylsulfanyl]benzoic acid, D-1b, was
prepared in a manner similar to that described in example A-1, step
(b): mp 131-135.degree. C.
[0224] (c) To a solution of 3-[(pyrazin-2-yl)methylsulfanyl]benzoic
acid, D-1b, (0.15 g, 0.61 mmol), 4-isopropyl-3-methylaniline
hydrochloride (0.113 g, 0.61 mmol), and triethylamine (0.09 mL,
0.65 mmol) in 2 mL DMF at room temperature was added EDC (0.116 g,
0.61 mmol). After stirring at room temperature for 24 hr, the
mixture was partitioned between ethyl acetate (30 mL) and brine (30
mL). The aqueous layer was extracted twice with ethyl acetate (30
mL) and the combined organics were washed twice with water (20 mL),
once with brine(25 mL), dried over sodium sulfate, and concentrated
to dryness. The residue was filtered through silica gel using 10%
methanol in chloroform and then purified by radial chromatography
with a 2 mm rotor using a gradient of 0% to 50% ethyl acetate in
hexanes as eluent to yield N-(4-isopropyl-3-methylphenyl)-3-[(-
pyrazin-2-yl)methylsulfanyl]benzamide, D-1, as a pale amber oil
(0.09 g, 35%): .sup.1H NMR (DMSO-d.sub.6) .delta. 10.10 (s, 1H),
8.68 (s, 1H), 8.57 (d, 1H, J=1.6 Hz), 8.51 (d, 1H, J=2.4 Hz), 7.92
(s, 1H), 7.76 (d, 1H, J=7.7 Hz), 7.59 (d, 1H, J=7.9 Hz), 7.55 (d,
1H, J=8.3 Hz), 7.52 (d, 1H, J=1.9 Hz), 7.46 (t, 1H, J=7.8 Hz), 7.21
(d, 1H, J=8.4 Hz), 4.50 (s, 2H), 3.11-3.05 (m, 1H), 2.30 (s, 3H),
1.18 (d, 6H, J=6.8 Hz). Anal. calc'd for
C.sub.22H.sub.23N.sub.3OS.0.4H.sub.2O: C, 68.68; H, 6.24; N, 10.92;
S, 8.33. Found: C, 68.86; H. 6.11; N, 10.70; S, 8.23
EXAMPLE D-2
N-(2-Methylquinolin-6-yl)-3-[(pyrazin-2-yl)methylsulfanyl]Benzamide
[0225] 138
[0226] Example D-2 was prepared in a similar manner to that
described for D-1, except that 6-amino-2-methylquinoline was used
in place of 4-isopropyl-3-methylaniline in step (c): mp
102-105.degree. C.; .sup.1H NMR (DMSO-d.sub.6) .delta. 10.53 (s,
1H), 8.69 (s, 1H), 8.57 (d, 1H, J=1.3 Hz), 8.52 (d, 1H, J=2.4 Hz),
8.46 (s, 1H), 8.22 (d, 1H, J=8.4 Hz), 7.80-7.97 (m, 2H), 7.91 (d,
1H, J=9.1 Hz), 7.82 (d, 1H, J=7.8 Hz), 7.63 (d, 1H, J=7.8 Hz), 7.51
(d, 1H, J=7.8 Hz), 7.40 (d, 1H, J=8.5 Hz), 4.52 (s, 2H), 2.65 (s,
3H). Anal. calc'd for C.sub.22H.sub.18N.sub.4OS.0.3H.su- b.2O.0.2
EtOAc: C, 66.87; H, 4.97; N, 13.68; S, 7.83. Found: C, 66.77; H,
5.18; N, 13.40; S, 7.61.
EXAMPLE D-3
N-(2-Methyl-quinolin-6-yl)-3-(pyridin-3-ylmethylsulfanyl)-benzamide
Dihydrochloride
[0227] 139
[0228] Example D-3 was prepared in a similar manner to that
described for D-1, except that 3-picolyl chloride was used in place
of 2-(bromomethyl)pyrazine in step (b), and
6-amino-2-methylquinoline was used in place of
4-isopropyl-3-methylaniline in step (c): HPLC R.sub.t=12.2 min.;
TLC R.sub.f=0.4 (5% methanol/chloroform); .sup.1H NMR (500 MHz,
DMSO-d.sub.6 w/ D.sub.2O) .delta. 8.95 (d, 1H, J=8.6 Hz), 8.77-8.73
(m, 2H), 8.66 (dd, 1H, J=1.1, 5.5 Hz), 8.41-8.33 (m, 2H), 8.20 (d,
1H, J=9.2 Hz), 7.97-7.83 (m, 4H), 7.66-7.53 (m, 2H), 4.51 (s, 2H),
2.94 (s, 3H); .sup.13C NMR (75 MHz, DMSO-d.sub.6 w/ D.sub.2O)
.delta. 167.8, 158.4, 146.8, 146.4, 144.5, 143.8, 140.5, 139.5,
136.7, 136.6, 136.4, 134.9, 131.7, 130.5, 130.4, 129.2, 128.4,
128.2, 126.0, 122.7, 119.0, 35.2, 22.3; MS (ESI) m/z 386
[M+H].sup.+. Anal. calc'd for C.sub.23H.sub.19N.sub.3O.sub.2S.2HCl
0.3 H.sub.2O: C, 59.56; H, 4.69; N, 9.06; S, 6.91. Found: C, 59.56;
H, 4.66; N, 9.00; S, 6.82.
EXAMPLE E-1
N-(2-methyl-quinolin-6-yl)-3-[{5-(phenylamino)-2-H-pyrazol-3-yl}methylsulf-
anyl]Benzamide
[0229] 140
[0230] (a) To a solution of 3-thiobenzoic acid (5.0 g, 32.4 mmol)
in 150 mL of acetone was added cesium carbonate (22.2 g, 68.1 mmol)
and 2-chloro-N-methoxy-N-methylacetamide (4.9 g, 35.7 mmol). After
stirring for 1 h, the reaction was quenched with dropwise addition
of 1N HCl. The reaction mixture was partitioned between 100 mL of
ethyl acetate and 50 m]L of 1N HCl, dried over sodium sulfate, and
evaporated. The residue was chromatographed on silica gel using 33%
hexane/66% ethyl acetate/1% acetic acid to afford
2-[(3-carboxyphenyl)sulfanyl]-N-methoxy-N-methylace- tamide, E-1a,
as a white solid 8.5 g (96%). .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 8.15 (t, 1H, J=1.5 Hz), 8.06 (br s, 1H), 7.92 (dt, 1H,
J=7.8, 1.2 Hz), 7.68 (dq, 1H, J=6.0, 1.2 Hz), 3.88 (s, 2H), 3.74
(s, 3H), 3.22 (s, 3H).
[0231] (b) To a solution of
2-[(3-carboxyphenyl)sulfanyl]-N-methoxy-N-meth- ylacetamide, E-1a,
(0.84 g, 3.1 mmol) and EDC (0.66 g, 3.4 mmol) in 10 mL of THF was
added 6-amino-2-methylquinoline (0.54 g, 3.4 mmol). After 3 h, the
solution was concentrated and the residue was purified by column
chromatography with 1:2 hexane/ethyl acetate to afford
3-[(N-methoxy-N-methylcarbamoyl)methylsulfanyl]-N-(2-methylquinolin-6-yl)-
benzamide, E-1b, as a white crystalline solid 0.89 g (72%): .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 8.50 (br s, 1H), 8.45 (d, 1H,
J=1.2 Hz), 8.05-7.97 (m, 3H), 7.73 (dd, 1H, J=7.8, 1.5 Hz), 7.57
(dt, 1H, J=13.4, 4.8 Hz), 7.56 (d, 1H, 4.5 Hz), 7.39 (t, 1H, J=4.5
Hz), 7.27 (d, 1H, J=4.8 Hz), 3.88 (s, 2H), 3.80 (s, 3H), 3.24 (s,
3H), 2.71 (s, 3H); MS (ESI): Calculated for
C.sub.21H.sub.21N.sub.3O.sub.3S (M+H.sup.+): 395. Found: 395. Anal.
calc'd for C.sub.20H.sub.24N.sub.2O.sub.6S.0.2H.sub.2O: C, 63.20;
H, 5.41; N, 10.53; S, 8.03. Found: C, 63.03; H, 5.32; N, 10.35; S,
7.92.
[0232] (c) To a solution of thioacetanilide (0.30 g, 1.95 mmol) in
10 mL of anhydrous THF at -78.degree. C. was added n-BuLi (1.56 mL,
3.89 mmol, 2.5 M in hexane) over a 5 min period. The mixture was
warmed to 0.degree. C. for 1 h, then recooled to -78.degree. C. To
this solution was added a solution of
3-[(N-methoxy-N-methylcarbamoyl)methylsulfanyl]-N-(2-methylqu-
inolin-6-yl)benzamide, E-1b, (0.35 g, 0.89 mmol) in 5 mL in THF,
and the resulting solution was warmed to 0.degree. C. After 1 h, a
solution of 1:1 methanol:acetic acid (1.0 mL) was added dropwise
over 1 min. The reaction solution was partitioned between 30 mL of
MTBE and extracted with 1N hydrochloric acid (2.times.20 mL) and
saturated brine (1.times.30 mL), and the organic layer was dried
over sodium sulfate and concentrated to give a yellow oil.
Purification using column chromatography with 3:1 hexane/ethyl
acetate afforded N-(2-methyl-quinolin-6-yl)-[2-oxo-3-phenylt-
hiocarbamoyl-propylsulfanyl]benzamide, E-1c, as a pale yellow foam
0.25 g (56%): .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.66 (br s,
1H), 8.26 (br s, 1H), 7.94-7.85 (3H, m), 7.81 (br s, 1H), 7.71-7.66
(m, 2H), 7.51 (br s, 1H), 7.45-7.41 (m, 1H), 7.26-7.21 (m 4H),
7.13-7.10 (m, 1H), 3.50 (br s, 2H), 2.67 (s, 3H), 2.03 (s, 2H); MS
(ESI): Calculated for C.sub.27H.sub.23N.sub.3O.sub.2S.sub.2
(M+H.sup.+): 486 Found: 486.
[0233] (d) To a solution containing
N-(2-methyl-quinolin-6-yl)-[2-oxo-3-ph-
enylthiocarbamoyl-propylsulfanyl]benzamide, E-1c, in 4 mL of
ethanol was added acetic acid (0.038 mL, 0.67 mmol) followed by
hydrazine monohydrate (0.032 mL, 0.63 mmol). The solution was
stirred for 2 h, then concentrated to give the crude product as an
amber oil. Purification by radial chromatography (1 mm silica
plate) with 90% ethyl acetate/10% methanol as eluant afforded a tan
solid. Precipitation of product from dichloromethane by dropwise
addition of hexane gave 0.12 g (58%) of
N-(2-methyl-quinolin-6-yl)-3-[{5-(phenylamino)-2-H-pyrazol-3-yl}methylsul-
fanyl]benzamide, E-1, as a white solid: mp 172-174.degree. C. HPLC
Rt=13.51 min.; .sup.1H NMR (500 MHz, Acetone-d.sub.6) .delta.
8.40-8.38 (m, 1H), 8.01 (d, 1H, J=8.5 Hz), 7.91 (s, 1H), 7.86 (dd,
1H, J=8.0, 1.2 Hz), 7.76 (t, 2H, J=9.5 Hz), 7.47 (d, 1H, J=8.0 Hz),
7.35 (t, 1H, J=7.5 Hz), 7.24 (d, 1H, J=8.5 Hz), 7.16 (br s, 2H),
7.02 (t, 2H, J=7.0 Hz), 6.59 (t, 1H, J=7.0 Hz), 5.73 (s, 1H), 4.18
(s, 2H), 2.52 (s, 3H); HRMS (FAB): Calculated for
C.sub.27H.sub.23N.sub.5OS (M+H.sup.+): 466.1702. Found: 466.1715.
Anal. calc'd for C.sub.27H.sub.23N.sub.5OS.0.5 CH.sub.2Cl.sub.2: C,
65.05; H, 4.76; N, 13.79; S, 6.31. Found C, 64.94; H, 4.72; N,
13.47; S, 6.51.
EXAMPLE E-2
N-(3,4,5-trimethoxyphenyl)-3-[2-(5-phenylamino-2H-pyrazol-3-yl)ethyl]Benza-
mide
[0234] 141
[0235] Example E-2 was prepared in a similar manner to that
described for E-1, except that 3,4,5-trimethoxyaniline was used in
place of 6-amino-2-methylquinoline in step (b): mp 67-69.degree. C.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.20 (br s, 1H), 7.75 (br
s, 1H), 7.64 (d, 1H, J=7.8 Hz), 7.42 (d, 1H, J=7.2 Hz), 7.29 (t,
1H, J=7.5 Hz), 6.96-6.91 (m, 3H), 6.83 (t, 1H, J=7.2 Hz), 5.89 (s,
1H), 4.05 (s, 2H), 3.80 (s, 3H), 3.75 (s, 6H); HRMS (FAB):
Calculated for C.sub.26H.sub.26N.sub.4O.su- b.4S (M+H.sup.+):
491.1753 Found: 491.1737. Anal. calc'd for
C.sub.26H.sub.26N.sub.4O.sub.4S.0.4 Et.sub.2O: C, 63.72; H, 5.81;
N, 10.77; S, 6.16. Found: C, 63.47; H, 5.88; N, 10.52; S, 6.34.
EXAMPLE F-1
3-[{5-((E)-2-(4-Hydroxy-3-methoxyphenyl)ethenyl)-2H-pyrazol-3-yl}-methylsu-
lfanyl]-N-(2-methylquinolin-6-yl)Benzamide
[0236] 142
[0237] (a) To 5 mL of ethanol was added
(E)-4-(4-hydroxy-3-methoxy-phenyl)- -but-3-en-2-one (0.50 g, 2.60
mmol), 1 g of 4 .ANG. molecular sieves, acetic acid (0.16 mL, 2.60
mmol), and hydrazine carboxylic acid t-butyl ester (0.34 g, 2.60
mmol). After stirring for 18 h, the mixture was filtered and the
filtrate was concentrated to give a crude product as a tan solid,
which was chromatographed on silica with 2:1 hexane/ethyl acetate
to furnish N-[3-(4-hydroxy-3-methoxy-phenyl)-1-methyl-(E)-2-prope-
nylidene]hydrazinecarboxylic acid t-butyl ester, F-1a, 0.68 g (86%)
as a white solid. TLC Rf=0.45 (40% hexane/60% ethyl acetate).
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.87 (br s, 1H),
7.11-7.08 (m, 2H), 6.98-6.52 (m, 2H), 6.89 (d, 1H, J=8.0 Hz), 5.82
(br s, 1H), 3.93 (s, 3H), 2.13 (s, 3H), 1.53 (s, 9H); LRFAB:
Calculated for C.sub.16H.sub.22N.sub.2O.sub.4 (M+H.sup.+): 307.
Found: 307.
[0238] (b) To a solution of
N-[3-(4-hydroxy-3-methoxy-phenyl)-1-methyl-(E)-
-2-propenylidene]-hydrazinecarboxylic acid t-butyl ester, F-1a,
(0.50 g, 1.63) in 5 mL of dichloromethane was added
diisopropylethylamine (64 mL, 3.92 mmol) and
chlorotriisopropylsilane (0.77 mL, 3.59 mmol). After 20 h, the
mixture was concentrated and the residue was partitioned between 30
mL of MTBE and saturated sodium bicarbonate (2.times.30 mL). The
organic layer was filter through 10 g of silica and concentrated to
afford a yellow oil. Purification was accomplished using
chromatotron with a 2 mm rotor eluting with 80% hexane/20% ethyl
acetate to give 0.48 g (65%) of
N-{3-[4-methoxy-3-(triisopropyl-silanyloxy)-phenyl]-1-methyl-(E)-2-propen-
ylidene]hydrazinecarboxylic acid t-butyl ester, F-1b, as a white
solid: TLC Rf=0.82 (60% hexane/40% ethyl acetate); .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.47 (br s, 1H), 7.02 (d, 1H, J=1.5 Hz),
6.94 (d, 1H, J=16.5 Hz), 6.87 (dd, 1H, J=8.3 Hz, 1.5 Hz), 6.82 (d,
1H, J=8.0 Hz), 6.77 (d, 1H, J=16.5 Hz), 3.81 (s, 3H), 2.02 (s, 3H),
1.50 (s, 9H), 1.29-1.23 (m, 3H), 1.09 (d, 18H, J=7.0 Hz); LRFAB:
Calculated for C.sub.25H.sub.42N.sub.2O.sub.4Si (M+H.sup.+): 463;
Found: 463.
[0239] (c) To a -78.degree. C. solution of
N-[3-[4-methoxy-3-(triisopropyl-
silanyloxy)-phenyl]-1-methyl-(E)-2-propenylidene]hydrazinecarboxylic
acid t-butyl ester, F-1b, (0.33 g, 0.72 mmol) in 10 mL of anhydrous
THF was added n-BuLi (0.61 mL, 3.89 mmol, 2.5 M in hexane) over 5
minutes. After the addition was complete, the mixture was warmed to
0.degree. C. for 1 h and then recooled to -78.degree. C. To the
resulting mixture was added a solution of
3-[(N-methoxy-N-methylcarbamoyl)methylsulfanyl]-N-(2-methylqu-
inolin-6-yl)benzamide, E-1b, (0.14 g, 0.36 mmol) in 5 mL in THF,
and the resulting mixture was warmed to 0.degree. C. After 1 h, the
reaction was cooled to -78.degree. C. and quenched by adding 1:1
acetic acid:methanol (1 mL). The reaction mixture was then
partitioned between 30 mL of MTBE and 1 N aq. HCl (2.times.20 mL).
The organic layer was dried over sodium sulfate and concentrated to
give a yellow oil. Purification by silica gel chromatography, with
2:1 hexane/ethyl acetate as eluant, afforded 0.12 g (43%) of
N.sup.2-[1-{4-methoxy-3-(triisopropylsilanyloxy)-phenyl)-6-{3-(2-
-methyl-quinolin-6-ylcarbamoyl)phenylsulfanyl}-5-oxo-(E)-hex-1-en-3-yliden-
e]-hydrazinecarboxylic acid t-butyl ester, F-1c, as a pale yellow
foam: TLC R.sub.f=0.50 (60% ethyl acetate/hexane); .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 8.64 (s, 1H), 8.12-8.10 (m, 2H), 8.03 (br
s, 1H), 7.90 (d, 1H, J=8.0 Hz), 7.77 (d, 2H, J=7.0 Hz), 7.46 (t,
1H, J=8.0 Hz), 7.31 (d, 1H, J=5.0 Hz), 6.91-6.78 (m, 5H), 3.92-3.60
(m, 4H), 2.73 (s, 3H), 1.54-1.43 (m, 3H), 1.20 (s, 9H), 1.07 (d,
18H, J=9.5 Hz).
[0240] (d) To a solution of 0.10 g (0.14 mmol) of
N.sup.2-[1-{4-methoxy-3--
(triisopropylsilanyloxy)-phenyl}-6-{3-(2-methyl-quinolin-6-ylcarbamoyl)phe-
nylsulfanyl}-5-oxo-(E)-hex-1-en-3-ylidene]-hydrazinecarboxylic acid
t-butyl ester, F-1c, in 5 mL of dichloromethane was added 5 mL of
trifluoroacetic acid. After 1 h, the reaction mixture was
concentrated, 5 mL of toluene added, and the solvent removed again
to give an amber oil. The residue was purified by silica gel
chromatography with 1:1:1 hexane/dichloromethane/ethyl acetate as
eluant to give 0.089 g (92%) of
3-[{5-{(E)-2-(3-methoxy-4-triisopropylsilanyloxyphenyl)ethenyl}-2H-pyrazo-
l-3-yl}methylsulfanyl]-N-(2-methylquinolin-6-yl)benzamide, F-1d, as
tan solid: TLC Rf=0.18 (50% ethyl acetate/hexane); .sup.1H NMR (300
MHz, Acetone-d.sub.6) .delta. 10.21 (br s, 1H), 8.95 (d, 1H, J=1.5
Hz), 8.71 (d, 1H, J=8.4 Hz), 8.43-8.31 (m, 3H), 8.10 (d, 1H, J=7.5
Hz), 7.88-7.82 (m, 2H), 7.85 (t, 1H, J=7.8 Hz), 7.41-7.19 (m, 4H),
7.11 (d, 1H, J=8.1 Hz), 6.69 (s, 1H), 4.57 (s, 2H), 4.10 (s, 3H),
3.05 (s, 3H), 1.57-1.47 (m, 3H), 1.34 (d, 18H, J=8.5 Hz); MS (ESI):
Calculated for C.sub.39H.sub.46N.sub.3OSi (M+H.sup.+): 678; Found:
678.
[0241] (e) To a solution of
3-[{5-{(E)-2-(3-methoxy-4-triisopropylsilanylo-
xyphenyl)ethenyl}-2H-pyrazol-3-yl}methylsulfanyl]-N-(2-methylquinolin-6-yl-
)benzamide, F-1d, (0.057 g, 0.084 mmol) in 5 mL THF was added
tetrabutylammonium fluoride (1M) in tetrahydrofuran (0.093 mL,
06092 mmol). After 3 h, the solution was concentrated and the
residue was partitioned between 20 mL of ethyl acetate and 20 mL of
1M phosphate buffer at 7.0 pH. The organic layer was filtered
through 10 g of silica with 50 mL dichloromethane and concentrated
to give the crude product as an amber oil. The residue was further
purified by radial chromatography on a 1 mm plate with 3:1
hexane/ethyl acetate as eluant. The purified product was dissolved
in 1 mL of dichloromethane and hexane was added dropwise to
precipitate 0.34 g (77%) of 3-[{5-((E)-2-(4-hydroxy-3-methoxy-
phenyl)ethenyl)-2H-pyrazol-3-yl
}methylsulfanyl]-N-(2-methylquinolin-6-yl)- benzamide, F-1, as a
white solid: HPLC Rt=13.12 min.; TLC Rf=0.30 (50% ethyl
acetate/hexane); .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 9.67 (br
s, 1H), 8.41 (br s, 1H), 8.03 (d, 1H, J=8.5 Hz), 7.89-7.87 (m, 2H),
8.79 (d, 1H, J=9.0 Hz), 7.71 (d, 1H, J=7.5 Hz), 7.33 (t, 1H, J=8.0
Hz), 7.25 (d, 1H, J=8.5 Hz), 7.01 (s, 1H), 6.78 (d, 1H, J=16.5 Hz),
6.83-6.78 (m, 2H), 6.67 (d, 1H, J=8 Hz), 6.29 (s, 1H), 4.19 (s,
2H), 3.74 (s, 3H), 2.53 (s, 3H). HRMS (FAB): Calculated for
C.sub.30H.sub.26N.sub.4O.sub.3S (M+H.sup.+): 655.0780 Found:
655.0804. Anal. calc'd for C.sub.30H.sub.26N.sub.4O.sub.3S.0.8
EtOAc: C, 67.23; H, 5.51; N, 9.45; S, 5.41. Found: C, 67.08; H,
5.60; N, 9.73; S, 5.45.
EXAMPLE F-2
3-[5-(2-(3,4-Dimethoxyphenyl)ethenyl)-2H-pyrazol-3-yl)methylsulfanyl]-N-(2-
-methylquinolin-6-yl)Benzamide
[0242] 143
[0243] Example F-2 was prepared in a similar manner to that
described for F-1, except that acetophenone was used in place of
4-(4-hydroxy-3-methoxy-phenyl)-but-3-en-2-one in step (a), and the
protection/deprotection steps (b) and (e) were not needed: mp
99-101.degree. C.; TLC Rf=0.50 (75% dichloromethane/25% ethyl
acetate); HPLC Rt=14.04 min.; .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 8.33 (br s, 1H), 8.21 (d, 1H, J=2.0 Hz) 7.95 (br s, 1H),
7.80 (d, 1H, J=8.5 Hz), 7.75 (d, 1H, J=9.1 Hz), 7.70 (d, 1H, J=7.5
Hz), 7.61 (d, 2H, J=7.5 Hz), 7.51 (dd, 1H, J=9.3, 2.5 Hz), 7.43 (d,
1H, J=8.0 Hz), 7.32-7.25 (m, 5H), 7.15 (d, 8.5 Hz), 6.57 (s, 1H),
4.26 (s, 2H), 2.61 (s, 3H). HRMS (FAB): Calculated for
C.sub.27H.sub.22N.sub.4OS (M+H.sup.+): 451.1593. Found:
451.1580.
[0244] Anal. calc'd for C.sub.27H.sub.22N.sub.4OS.0.8 EtOAc: C,
70.42; H, 5.30; N, 11.33; S, 6.48. EtOAc Found: C, 70.39; H, 5.34;
N, 11.29; S, 6.48.
EXAMPLE F-3
3-(2-{5-[(E)-2-(3,4-Dimethoxyphenyl)ethenyl]-2H-pyrazol-3-yl}-ethyl)-N-(3--
methyl-4-isopropylphenyl)-benzamide
[0245] 144
[0246] Example F-3 was prepared in a similar manner to that
described for F-1, except that
(E)-4-(3,4-dimethoxyphenyl)-but-3-en-2-one was used in place of
(E)-4-(4-hydroxy-3-methoxyphenyl)-but-3-en-2-one in step (a),
N-(4-isopropyl-3-methyl-phenyl)-3-[2-(N-methoxy-N-methylcarbamoyl)-ethyl]-
benzamide, G-1f, (from Example G-1, step (f)) was used in place of
3-[(N-methoxy-N-methylcarbamoyl)methylsulfanyl]-N-(2-methylquinolin-6-yl)-
benzamide, E-1b, in step (c) and the protection/deprotection steps
(b) and (e) were not needed: HPLC Rt=16.37 min.; .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 7.71 (br s, 1H), 7.69-7.66 (m, 2H),
7.44-7.17 (m, 5H), 7.01-6.81 (m, 5H), 6.25 (s, 1H), 3.89 (d, 6H,
J=1.5 Hz), 3.12-3.01 (m, 5H), 2.32 (s, 3H), 1.20 (d, 6H, J=5.1 Hz);
HRMS (FAB): Calculated for m/z C.sub.32H.sub.35N.sub.3O.sub.3
(M+Cs.sup.+): 642.1733, Found: 642.1712. Anal. calc'd for
C.sub.32H.sub.35N.sub.3O.sub.3: C, 75.42; H, 6.92; N, 8.24. Found
C, 75.45; H, 7.08; N, 8.16.
EXAMPLE F-4
4-Fluoro-3-[{5-((E)-1-propenyl)-2H-pyrazol-3-yl}methoxy]-N-[4-(pyrrolidin--
1-yl)-3-trifluoromethylphenyl]Benzamide
[0247] 145
[0248] Example F-4 was prepared in a similar manner to that
described for F-1, except that (E)-3-penten-2-one was used in place
of (E)-4-(4-hydroxy-3-methoxyphenyl)-but-3-en-2-one in step (a),
4-fluoro-N-[4-(pyrrolidin-1-yl)-3-trifluoromethylphenyl]-3-[2-(N-methoxy--
N-methylcarbamoyl)methoxy]benzamide, (prepared in a similar manner
as described for
4-fluoro-N-[4-(imidazol-1-yl)-3-trifluoromethyl-phenyl]-3-[-
(N-methoxy-N-methylcarbamoyl)methoxy]benzamide, J-1d, in Example
J-1) was used in place of
3-[(N-methoxy-N-methylcarbamoyl)methylsulfanyl]-N-(2-met-
hylquinolin-6-yl)benzamide, E-1b, in step (c) and the
protection/deprotection steps (b) and (e) were not needed: HPLC
Rt=16.27 min. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.75 (m,
4H), 7.45 (d, 1H, J=8.7 Hz), 7.42-7.37 (m, 1H), 7.35-7.28 (m, 2H),
7.15 (t, 1H, J=8.4 Hz), 7.00 (d, 1H, J=8.7 Hz), 6.56 (d, 1H, J=8.9
Hz), 6.37 (s, 1H), 6.27-6.15 (m, 2H), 5.60 (s, 1H), 5.23 (s, 1H),
3.30-3.29 (m, 4H), 1.98-1.95 (m, 4H), 1.92 (d, 3H, J=6.3 Hz); MS
(ESI): m/z Calculated for C.sub.21H.sub.24F.sub.4N.sub.4O.sub.2
(M+H.sup.+): 489, Found:489.
[0249] Anal. calc'd for C.sub.25H.sub.24F.sub.4N.sub.4O.sub.2: C,
61.47; H, 4.95; N, 11.47. Found C, 61.32; H, 5.06; N 11.33.
EXAMPLE F-5
3-(2-{5-[(E)-2-(3,4-Dimethoxyphenyl)ethenyl]-2H-pyrazol-3-yl}-ethyl)-N-(3--
methyl-4-isopropylphenyl)-benzamide
[0250] 146
[0251] Example F-5 was prepared in a similar manner to that
described for F-3, except that (E)-4-(pyridin-3-yl)-3-buten-2-one
was used in place of (E)-4-(3,4-dimethoxyphenyl)-but-3-en-2-one in
step (a), and
4-[4-(t-butoxycarbonyl)piperazin-1-yl]-3-trifluoromethylaniline,
prepared according to the procedure described in WO 99/21845 (p.
58) for the preparation of 1-methyl-4-(4-nitrophenyl)piperazine,
was used in place of 4-isopropyl-3-methylaniline in step (d) of
Example G-1, and the final deprotection step (e) was carried out in
a manner similar to that described in Example G-10: HPLC Rt=13.53
min.; .sup.1H NMR (300 MHz, DSMO-d.sub.6) .delta. 8.69 (s, 1H),
8.44 (d, 1H, J=3.3 Hz), 8.14 (s, 1H), 8.07-7.82 (m, 2H), 7.86 (s,
1H), 7.81-7.79 (m, 1H), 7.54-7.43 (m, 3H), 7.39-7.37 (m, 1H),
7.25-7.04 (m, 2H), 310-2.98 (m, 4H), 2.83-2.78 (m, 8H); HRMS (FAB):
Calculated for C.sub.30H.sub.29F.sub.3N.sub.6O (M+H.sup.+):
547.2433 Found: 547.2445. Anal. calc'd for
C.sub.28H.sub.30N.sub.4O.0.5H.sub.2O.0.2 CH.sub.2Cl.sub.2: C,
63.35; H 5.35; N, 14.68. Found C, 63.26; H, 5.38; N, 14.25.
EXAMPLE G-1
N-(4-Isopropyl-3-methyl-phenyl)-3-{2-[5-(4-(methylsulfamoyl)-phenylamino)--
2H-pyrazol-3-yl]-ethyl}-benzamide
[0252] 147148
[0253] (a) To a solution of 4-acetylamino-benzenesulfonyl chloride
(2.00 g, 8.56 mmol) in 25 mL of DMF was added
4-dimethylaminopyridine (0.11 g, 0.86 mmol) and
diphenylaminomethane (1.79 mL, 10.27 mmol). After 4 h, the reaction
mixture was added to 125 mL of water, and the precipitate was
collected by filtration, washed with diethyl ether (2.times.20 mL),
then dried under high vacuum for 4 h to afford 1.98 g (61%) of
N-(diphenylmethyl)-4-(acetylamino)benzenesulfonamide, G-1a: HPLC
Rt=12.90 min.; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 10.20 (br
s, 1H), (8.82 (d, 1H, J=2.0 Hz), 7.57 (s, 4H), 7.24-7.20 (m, 8H),
7.18-7.13 (m, 2H), 5.58-7.52 (m, 1H), 3.38 (s, 3H); LRFAB:
Calculated for C.sub.21H.sub.20N.sub.2O.sub.3S (M+H.sup.+): 381
Found:381.
[0254] (b) To DMF (15 mL) was added
N-(diphenylmethyl)-4-(acetylamino)benz- enesulfonamide, G-1a, (1.50
g, 3.94 mmol), potassium carbonate (1.37 g, 9.86 mmol), and
iodomethane (0.37 mL, 5.93 mmol). The reaction mixture was stirred
for 3 h at 60.degree. C., cooled to 25.degree. C., and partitioned
between 50 mL of MTBE and 1N HCl (2.times.50 mL). The organic layer
was dried over sodium sulfate and concentrated to give an amber
oil, which was purified by chromatography on silica gel (3:1
hexane:ethyl acetate) to give 1.50 g (98%) of
N-(diphenylmethyl)-N-methyl-4-(acetylami- no)benzenesulfonamide,
G-1b: TLC Rf=0.62 40% hexane/ethyl acetate; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.67-7.64 (m, 1H), 7.58-7.52 (m, 2H), 7.28-7.23
(m, 8H), 7.21-7.08 (m, 3H), 6.47 (s, 1H), 2.68 (s, 3H), 2.20 (s,
3H).
[0255] (c) To a solution of 2.00 g, (5.30 mmol) of
N-(diphenylmethyl)-N-me- thyl-4-(acetylamino)-benzenesulfonamide,
G-1b, in toluene (30 mL) was added
2,4-bis(4-methoxyphenyl)-1,3-dithia-2,4-diphosphetane-2,4-disulfide
(1.07 g, 2.64 mmol). The solution was warmed to 100.degree. C. for
2 h then cooled to 25.degree. C. The reaction mixture was filtered
through a silica gel plug using 50 mL of MBTE and concentrated. The
residue was purified by chromatography on silica gel (5:1
hexane/ethyl acetate) to afforded 1.62 g (77%) of
4-[N-(diphenylmethyl)-N-methylsulfamoyl]-thioace- tanilide, G-1c,
as a yellow oil: HPLC Rt=18.67 min.; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.84 (d, 1H, J=8.7 Hz), 7.72 (d, 1H, J=8.5 Hz),
7.28-7.25 (m, 8H), 7.09-7.07 (m, 2H), 6.46 (s, 1H), 2.77 (s, 3H),
2.56 (s, 3H).
[0256] (d) To a solution of 5.00 g (33.26 mmol) 3-formylbenzoic
acid in dichloromethane at 0.degree. C. was added oxalyl chloride
(3.48 mL, 39.92 mmol) and DMF (0.01 mL). The reaction was stirred
for 3 h at 25.degree. C., and then concentrated to dryness. The
residue (2.1 g, 12.45 mmol) was dissolved in dichloromethane (30
mL) and 4-isopropyl-3-methylaniline.HCl (2.55 g, 13.70 g) added,
followed by diisopropylethylamine (4.48 mL). After stirring for 2
h, the solution was washed with sat. sodium bicarbonate (2.times.20
mL) and 1N HCl (2.times.20 mL) and the organic layer dried over
sodium sulfate and concentrated to dryness. The residue was
purified on silica gel (3:1 hexane:ethyl acetate) to provide 2.99 g
(90%) of 3-formyl-N-(4-isopropyl-3-methylphenyl)benzamide, G-1d, as
a off-white solid: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 10.12
(s, 1H), 8.60-8.58 (m, 1H), 8.38-8.36 (m, 1H), 8.21-8.15 (m, 2H),
8.07-8.04 (m, 1H), 7.93 (br s, 1H), 7.71-7.67 (m, 1H), 7.44 (br s,
1H), 7.27-7.24 (s, 1H), 3.15-3.10 (m, 1H), 2.35 (s, 3H), 1.23 (d,
6H, J=6.7 Hz).
[0257] (e) To a solution of 2-diethyl (N-methoxy-N
methyl-carbamoylmethyl)- phosphonate (1.70 mL, 7.82 mmol) in 15 ml
of THF at -78.degree. C. was added sodium bis(trimethylsilyl)amide
(9.24 mL, 1M in THF) dropwise over 2 min. After the addition was
complete, the reaction was warmed to 0.degree. C. for 1 h. To this
solution was added a solution of 2.00 g (7.11 mmol) of
3-formyl-N-(4-isopropyl-3-methyl-phenyl)-benzamide, G-1d, in 20 mL
of THF. The solution was stirred for 1 h, then quenched with 1:1
MeOH/AcOH (1 mL). The mixture was partitioned between 50 mL of
ethyl acetate and 1N HCl (2.times.20 mL), and the organic layer was
dried over sodium sulfate and concentrated. The residue was
purified on silica gel (2:1 hexane:ethyl acetate) and to afford
1.33 g (51%) of
N-(4-isopropyl-3-methylphenyl)-3-[(E)-2-(N-methoxy-N-methylcarbamoyl)ethe-
nyl]-benzamide, G-1e, as a white solid foam: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.10 (br s, 1H), 7.83-7.79 (m, 2H), 7.73-7.68
(m, 2H), 7.52-7.45 (m, 3H), 7.28-7.24 (m, 1H), 7.12 (d, 1H, J=15.9
Hz), 3.78 (s, 3H), 3.31 (s, 3H), 3.15-3.10 (m, 1H), 2.36 (s, 3H),
1.23 (d, 6H, J=6.0 Hz).
[0258] (f) A mixture of 0.90 g (2.46 mmol)
N-(4-isopropyl-3-methylphenyl)--
3-[(E)-2-(N-methoxy-N-methylcarbamoyl)ethenyl]-benzamide, G-1e, and
0.1 g of 10% palladium on carbon in 20 ml of 1:1 MeOH:EtOAc was
stirred under 1 atm H.sub.2 for 18 h. The reaction was filtered
though a 0.5 uM teflon filter and concentrated to give 0.90 g
(100%) of N-(4-isopropyl-3-methylp-
henyl)-3-[2-(N-methoxy-N-methylcarbamoyl)ethyl]-benzamide, G-1f:
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.74 (s, 2H), 7.68 (d,
1H, J=4.2 Hz), 7.44-7.40 (m, 4H), 7.25-7.23 (m, 1H), 3.63 (s, 3H),
3.18 (s, 3H), 3.14-3.11 (m, 1H), 3.06-3.03 (m, 2H), 2.79 (t, 2H,
J=4.5 Hz), 2.36 (s, 3H), 1.22 (d, 6H, J=4.2 Hz).
[0259] (g) To a solution of 0.45 g (1.34 mmol) of
4-[N-(diphenylmethyl)-N-- methylsulfamoyl]thioacetanilide, G-1c,
and 0.16 mL (1.34 mmol) of N,N'-dimethylpropyleneurea (DMPU) in 15
mL of THF at -78.degree. C. was added 1.07 mL (2.68 mmol) of 2.5 M
n-BuLi in hexane. After 0.25 h, the reaction was warmed to
0.degree. C. for 0.5 h, then recooled to -78.degree. C. To the
reaction mixture was added a solution of 0.23 g (0.62 mmol) of
N-(4-isopropyl-3-methylphenyl)-3-[2-(N-methoxy-N-methylcar-
bamoyl)ethyl]-benzamide, G-1f, in 5 mL of THF. The reaction was
warmed to 0.degree. C. for 1 hour and then quenched by dropwise
addition of 0.5 mL of 1:1 MeOH/AcOH. The mixture was partitioned
between 30 mL of ethyl acetate and sat. sodium bicarbonate
(2.times.10 mL). The organic layer was dried over sodium sulfate
and concentrated. The residue was purified by chromatography on
silica gel (1:2 ethyl acetate:hexane) to afford 0.24 g (54%) of
3-[4-{4-(N-(diphenylmethyl)-N-methylsulfamoyl)phenylthiocarbam-
oyl}-3-oxo-butyl]-N-(4-isopropyl-3-methylphenyl)benzamide, G-1 g,
as an amber oil: .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.91-7.83 (m, 2H), 7.74-7.64 (m, 3H), 7.59-7.47 (m, 3H), 7.41-7.23
(m, 8H) 7.12-7.04 m, 6H), 6.43 (s, 1H), 7.25-7.23, (m, 1H), 3.63
(s, 3H), 3.18 (s, 3H), 3.18-3.13 (m, 1H), 3.11-3.01 (m, 2H), 2.83
(s, 2H), 2.78 (s, 3H), 2.37-2.32 (m, 2H), 1.21 (d, 6H, 1.5 Hz).
[0260] (h) To a solution of 0.36 g (0.51 mmol) of
3-[4-{4-(N-(diphenylmeth-
yl)-N-methyl-sulfamoyl)phenylthiocarbamoyl}-3-oxo-butyl]-N-(4-isopropyl-3--
methylphenyl)benzamide, G-1 g, in 5 ml of ethanol were added acetic
acid (0.20 mL, 0.34 mmol) and hydrazine monohydrate (0.016 mL, 0.34
mmol). After 1 h, the solution was partitioned between ethyl
acetate (30 mL) and 1N HCL (2.times.20 mL). The organic layer was
washed with sat. sodium bicarbonate (2.times.20 mL), dried over
sodium sulfate, and concentrated to give a yellow oil. Purification
by chromatography on silica gel (25% ethyl acteate:hexane) afforded
0.28 g (79%) of 3-{2-[5-(4-(N-(diphenylmet-
hyl)-N-methylsulfamoyl)phenylamino}-2H-pyrazol-3-yl]ethyl}-N-(4-isopropyl--
3-methyl-phenyl)-benzamide, G-1 h, as yellow solid: .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.09 (s, 1H), 7.67 (br s, 1H), 7.63
(d, 1H, J=7.5 Hz), 7.48-7.42 (m, 3H), 7.60-7.22 (m, 10H), 7.01-7.04
(m, 3H), 7.01 (d, 2H, J=8.4 Hz), 6.43 (s, 1H), 5.78 (s, 1 h),
3.13-3.04 (m, 1H), 2.93-2.84 (m, 4H), 2.61 (s, 3H), 2.28 (s, 3H),
1.19 (d, 6H, J=6.6 Hz); MS (ESI): m/z Calculated for
C.sub.42H.sub.43N.sub.5O.sub.3S (M-H.sup.-): 696 Found 696.
[0261] (i) To a solution of
3-{2-[5-{4-(N-(diphenylmethyl)-N-methylsulfamo-
yl)phenylamino}-2H-pyrazol-3-yl]ethyl}-N-(4-isopropyl-3-methyl-phenyl)-ben-
zamide, G-1 h, in 0.025 mL of formic acid and 1.2 mL of acetic acid
was added 0.08 goof 10% palladium on carbon. The mixture was
stirred at 80.degree. C. for 96 h, then cooled to room temperature
and filtered though a 0.5 .mu.M Teflon filter. The filtrate was
concentrated and the residue was purified by chromatography on
silica gel (2:1 ethyl acetate:hexane) to afford
N-(4-isopropyl-3-methyl-phenyl)-3-{2-[5-(4-(N-m-
ethylsulfamoyl)phenylamino)-2H-pyrazol-3-yl]ethyl}benzamide, G-1,
(0.053 g, 87%): HPLC Rt=14.65 min; .sup.1H NMR (Acetone-d.sub.6)
.delta. 791 (br s, 1H), 7.59 (br s, 1H), 7.69 (d, 1H, J=6.0 Hz),
7.52-7.25 (m, 7H), 7.08 (d, 1H, J=8.1 Hz), 3.51-2.89 (m, 5H), 2.40
(s, 3H), 2.19 (s, 3H), 1.08 (d, 6H, J=6.9 Hz); HRMS (FAB):
Calculated for C.sub.29H.sub.33N.sub.5O.su- b.3S (M+H.sup.+):
532.2382, Found: 532.2891
EXAMPLE G-2
N-(2-Methylquinolin-6-yl)-3-[2-(5-phenylamino-2H-pyrazol-3-yl)ethyl]Benzam-
ide
[0262] 149
[0263] Example G-2 was made in a similar manner to that described
for G-1, except that 6-amino-2-methylquinoline was used in place of
4-isopropyl-3-methylaniline in step (d), and thioacetanilide was
used instead of
4-[N-(diphenylmethyl)-N-methylsulfamoyl]thioacetanilide, G-1c, in
step (g), and the final deprotection step (i) was not needed: mp
98-102.degree. C. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.79
(d, 1H, J=2 Hz), 8.32 (br s, 1H), 7.95 (d, 1H, J=8.5 Hz), 7.92 (d,
1H, J=9.0 Hz), 7.22 (d, 1H, J=7.5 Hz), 6.74 (dd, 1H, J=9.0, 2.0
Hz), 7.37 (t, 1H, J=7.5 Hz), 7.31-7.20 (m, 6H), 7.06 (d, 2H, J=8.0
Hz), 6.85 (t. 1H, J=7.5 Hz), 5.81 (s, 1H), 2.96 (t, 2H, J=6.0 Hz),
2.91 (t, 2H, J=6.0 Hz) 2.65 (s, 3H); HRMS (FAB): Calculated for
C.sub.28H.sub.25N.sub.5O (M+H.sup.+): 448.2137 Found:448.2129.
Anal. calc'd for C.sub.28H.sub.25N.sub.5O.0.3 EtOAc: C, 73.99; H,
5.83; N, 14.78. Found C, 73.72; H, 5.88; N, 14.78.
EXAMPLE G-3
N-(4-isopropyl-3-methylphenyl)-3-[2-(5-phenylamino-2H-pyrazol-3-yl)ethyl]B-
enzamide
[0264] 150
[0265] Example G-3 was made in a similar manner to that described
for G-1, except thioacetanilide was used instead of
4-[N-(diphenylmethyl)-N-methyl- sulfamoyl]thioacetanilide, G-1c, in
step (g), and the final deprotection step (i) was not needed: mp
150-151.degree. C.; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.84
(s, 1H), 7.65 (d, 1H, J=7.5 Hz), 7.62 (s, 1H), 7.42-7.35 (m, 3H),
7.36 (t, 1H, J=7.5 Hz), 7.28-7.17 (m, 4H), 7.05 (d, 2H, J=8.5 Hz),
5.80 (s, 1H), 3.12-3.07 (m, 1H), 2.97 (t, 2H, J=7.5 Hz), 2.90 (t,
2H, J=7.0 Hz), 2.30 (s, 3H); HRMS (FAB): Calculated for
C.sub.28H.sub.30N.sub.4O (M+H.sup.+): 439.2498 Found: 439.2488.
Anal. calc'd for C.sub.28H.sub.30N.sub.4O.0.1CH.sub.2Cl.sub.2
Found: C, 75.49; H 6.81; N, 12.53. Found C, 75.44; H, 6.81; N,
12.53.
EXAMPLE G-4
N-(4-Isopropyl-3-methyl-phenyl)-3-{2-[5-(6-methoxypyridin-3-yl)Amino-2H-py-
razol-3-yl]-ethyl}-benzamide
[0266] 151
[0267] Example G-4 was made in a similar manner to that described
for G-1, except that N-(6-methoxy-pyridin-3-yl)acetamide was used
instead of
N-(diphenylmethyl)-N-methyl-4-(acetylamino)-benzenesulfonamide,
G-1b, in step (c). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.97
(d, 1H, J=2.7 Hz), 7.84 (br s, 1H), 7.67-7.62 (m, 2H), 7.49 (dd,
1H, J=8.9, 3.0 Hz), 7.42-7.40 (m, 3H), 7.30-7.61 (m, 2H), 7.19 (d,
1H, J=9.0 Hz), 5.66 (s, 1H), 3.88 (s, 3H), 3.10 (q, 1H, J=6.6 Hz),
2.97-2.90 (m, 4H), 2.17 (s, 3H), 1.23 (d, 6H, J=4.2 Hz); HRMS
(FAB): Calculated for C.sub.28H.sub.31N.sub.5O.sub.2 (M+H.sup.+):
470.2556, Found: 470.2563. Anal. calc'd for
C.sub.28H.sub.33N.sub.5O.sub.2Cl.sub.2: C, 61.99; H, 6.13; N,
12.91. Found C, 61.83; H, 6.39; N, 12.83.
EXAMPLE G-5
N-(4-Dimethylamino-3-trifluoromethylphenyl)-3-{2-[5-(6-methoxypyridin-3-yl-
)amino-2H-pyrazol-3-yl]ethyl}-benzamide
[0268] 152
[0269] Example G-5 was made in a similar manner to that described
for G-1, except that 4-(dimethylamino)-3-trifluoromethylaniline,
prepared according to the procedure described in WO 99/21845 (p.
58) for the preparation of 1-methyl-4-(4-nitrophenyl)piperazine,
was used in place of 4-isopropyl-3-methylaniline in step (d), and
N-(6-methoxy-pyridin-3-yl)ac- etamide was used instead of
N-(diphenylmethyl)-N-methyl-4-(acetylamino)-be- nzenesulfonamide,
G-1b, in step (c): .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.27
(br s, 1H), 7.93 (d, 1H, J=1.5 Hz), 7.84 (dd, 1H, J=4.5, 1.5 Hz),
7.88 (d, 1H, J=2.7 Hz), 7.64 (d, 1H, J=5.7 Hz), 7.60 (s, 1H), 7.37
(dd, 1H, J=8.7, 3.0 Hz), 7.66 (d, 1H, 4.5 Hz), 7.58 (s, 1H), 7.43
(dd, 1H, J=5.40, 1.5 Hz), 7.33 (t, 1H, J=4.8 Hz), 7.29-7.26 (m,
2H), 6.63 (d, 1H, J=5.4 Hz), 5.62 (s, 1H) 3.86 (s, 3H), 2.94-2.86
(m, 4H), 2.69 (s, 6H); HRMS (FAB): Calculated for
C.sub.27H.sub.27F.sub.3N.sub.6O.sub.2 (M+H.sup.+): 525.2226, Found:
525.2208. Anal. calc'd for
C.sub.27H.sub.27F.sub.3N.sub.6O.0.5H.sub.2O: C, 60.78; H, 5.29; N,
15.75. Found C, 61.15; H, 5.25; N, 15.7
EXAMPLE G-6
N-(6-Dimethylamino-5-trifluoromethylpyridin-3-yl)-3-{2-[5-(6-methoxypyridi-
n-3-yl)amino-2H-pyrazol-3-yl]ethyl}-benzamide
[0270] 153
[0271] Example G-6 was made in a similar manner to that described
for G-1, except that
5-amino-2-(dimethylamino)-3-trifluoromethylpyridine, prepared
according to the procedure described in WO 99/21845 (p. 58) for the
preparation of 1-methyl-4-(4-nitrophenyl)piperazine, was used in
place of 4-isopropyl-3-methylaniline in step (d), and
N-(6-methoxy-pyridin-3-yl)ac- etamide was used instead of
N-(diphenylmethyl)-N-methyl-4-(acetylamino)-be- nzenesulfonamide,
G-1b, in step (c): HPLC Rt=14.04 min; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.73 (br s, 1H), 8.47 (d, 1H, J=2.4 Hz), 8.18
(d, 1H, J=2.0 Hz), 7.88 (d, 1H, J=2.7 Hz), 7.64 (d, 1H, J=5.7 Hz),
7.60 (s, 1H), 7.37 (dd, 1H, J=8.7, 3.0 Hz), 7.29-7.21 (m, 2H), 6.58
(d, 1H, J=8.7 Hz), 5.78 (s, 1H), 3.83 (s, 3H), 2.94 (s, 6H),
2.86-2.76 (m, 4H); HRMS (FAB): Calculated for
C.sub.26H.sub.26F.sub.3N.sub.7O.sub.3 (M+H.sup.+): 526.2178, Found:
526.2194. Anal. calc'd for
C.sub.26H.sub.26F.sub.3N.sub.7O.sub.3.0.4 Et.sub.2O: C, 59.71; H,
5.45; N, 17.66. Found C, 59.58; H, 5.44; N, 17.53.
EXAMPLE G-7
N-(3,5-Dichloro-4-dimethylaminophenyl)-3-{2-[5-(6-methoxy-pyridin-3-yl)ami-
no-2H-pyrazol-3-yl]ethyl}Benzamide
[0272] 154
[0273] Example G-7 was made in a similar manner to that described
for G-1, except that 3,5-dichloro-4-(pyrrolidino)aniline, prepared
according to the procedure described in WO 99/21845 (p. 58) for the
preparation of 1-methyl-4-(4-nitrophenyl)piperazine, was used in
place of 4-isopropyl-3-methylaniline in step (d), and
N-(6-methoxy-pyridin-3-yl)ac- etamide was used instead of
N-(diphenylmethyl)-N-methyl-4-(acetylamino)-be- nzenesulfonamide,
G-1b, in step (c): HPLC Rt=16.07 min; .sup.1H NMR (300 MHz,
Acetone-d.sub.6) .delta. 8.25 (d, 1H, J=2.7 Hz), 7.91 (br s, 2H),
7.87 (s, 1H), 7.82-7.78 (m, 2H), 7.45-7.38 (m, 3H), 6.61 (d, 1H,
J=9.3H), 5.62 9 s, 1H), 3.78 (s, 3H), 3.31-2.97 (m, 4H), 2.84 (m,
6H); HRMS (FAB): Calculated for C.sub.26H.sub.26N.sub.6O.sub.2
(M+H.sup.+): 525.1573, Found: 525.1559. Anal. calc'd for
C.sub.26H.sub.26N.sub.6O.sub.23.0.3 Et.sub.2O: C, 59.65; H, 5.34;
N, 15.35. Found C, 59.35; H, 5.25; N, 17.35.
EXAMPLE G-8
3-{2-[5-(6-Methoxypyridin-3-yl)amino-2H-pyrazol-3-yl]-ethyl}-N-(4-pyrrolid-
in-1-yl-3-trifluoromethylphenyl)Benzamide
[0274] 155
[0275] Example G-8 was made in a similar manner to that described
for G-1, except that 4-(pyrrolidin-1-yl)-3-trifluoromethylaniline,
prepared according to the procedure described in WO 99/21845 (p.
58) for the preparation of 1-methyl-4-(4-nitrophenyl)piperazine,
was used in place of 4-isopropyl-3-methylaniline in step (d), and
N-(6-methoxy-pyridin-3-yl)ac- etamide was used instead of
N-(diphenylmethyl)-N-methyl-4-(acetylamino)-be- nzenesulfonamide,
G-1b, in step (c): HPLC Rt=15.66 min; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.35 (br s, 1H), 7.91 (d, 1H, J=1.5 Hz), 7.70
(d, 1H, J=1.2 Hz), 7.64 (dd, 2H, J=12.5, 4.5 Hz), 7.57 (s, 1H),
7.38 (dd, 1H, J=5.3, 1.5 Hz), 7.27-7.20 (m, 1H), 6.90 (d, 1H, J=5.4
Hz), 6.59 (d, 1H, J=5.1 Hz), 5.60 (s, 1H), 3.84 (s, 3H), 3.25 (t,
4H, 3.6 Hz), 2.88-2.76 (m, 4H), 1.94-1.90 (m, 4H); HRMS (FAB):
Calculated for C.sub.29H.sub.29F.sub.3N.sub.6O.sub.2 (M+H.sup.+):
551.2382, Found: 551.2389. Anal. calc'd for
C.sub.29H.sub.29F.sub.3N.sub.6O.sub.2.0.2CH.su- b.2Cl.sub.2: C,
61.09; H, 5.18; N, 14.59. Found C, 61.36; H, 5.18; N, 14.59.
EXAMPLE G-9
3-{2-[5-(6-Methoxypyridin-3-yl)amino-2H-pyrazol-3-yl]-ethyl}-N-[4-(4-t-but-
oxycarbonylpiperazin-1-yl)-3-trifluoromethylphenyl]Benzamide
[0276] 156
[0277] Example G-9 was made in a similar manner to that described
for G-1, except that
4-[4-(t-butoxycarbonyl)piperazin-1-yl]-3-trifluoromethylanili- ne;
prepared according to the procedure described in WO 99/21845 (p.
58) for the preparation of 1-methyl-4-(4-nitrophenyl)piperazine,
was used in place of 4-isopropyl-3-methylaniline in step (d), and
N-(6-methoxy-pyridin-3-yl)acetamide was used instead of
N-(diphenylmethyl)-N-methyl-4-(acetylamino)-benzenesulfonamide,
G-1b, in step (c): .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.27
(br s, 1H), 7.95-7.87 (m, 3H), 7.70 (d, 1H, J=7.5 Hz), 7.59 (br s,
1H), 7.57 (s, 1H), 7.47 (dd, 1H, J=8.7, 3.0 Hz), 7.43-7.32 (m, 2H),
7.29-7.21 (m, 2H), 7.28-7.26 (m, 2H), 6.66 (d, 1H, J=8.7 Hz), 5.60
(s, 1H), 3.88 (s, 3H), 3.56-3.51 (m, 4H), 3.26-3.24 (m, 4H),
3.00-2.93 (m, 4H), 2.83-2.82 (m, 4H), 1.49 (s, 9H); HRMS (FAB):
Calculated for C.sub.34H.sub.38F.sub.3N.su- b.7O.sub.4
(M+Na.sup.+): 688.2835, Found: 688.2856.
[0278] Anal. calc'd for
C.sub.34H.sub.38F.sub.3N.sub.7O.sub.4.0.4H.sub.2O: C, 60.68; H,
5.81; N, 14.57. Found C, 60.90; H, 5.88; N, 14.57.
EXAMPLE G-10
3-{2-[5-(6-Methoxypyridin-3-yl)amino)-2H-pyrazol-3-yl]ethyl}-N-(4-piperazi-
n-1-yl-3-trifluoromethylphenyl)Benzamide
[0279] 157
[0280] To a solution of 0.075 g (0.113 mmol) of
3-{2-[5-(6-Methoxypyridin--
3-yl)amino-2H-pyrazol-3-yl]-ethyl}-N-[4-(4-t-butoxycarbonyl)piperazin-1-yl-
-3-trifluoromethylphenyl]benzamide, G-9, in 5 mL of dichloromethane
was added 5 mL of trifluoroacetic acid. After 2 h, the reaction
mixture was concentrated. The residue was dissolved in 20 mL
chloroform/isopropanol (10:1) and washed with sat. aq. sodium
bicarbonate (2.times.10 mL). The organic layer was filtered though
a silica plug using ethanol to elute product and concentrated to
afford 3-{2-[5-(6-methoxypyridin-3-yl)amino)--
2H-pyrazol-3-yl]ethyl}-N-(4-piperazin-1-yl-3-trifluoromethylphenyl)benzami-
de, G-10, as off-white solid (0.056 g, 88%): HPLC Rt=12.95 min.;
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 11.38 (br s, 1H), 8.51
(br s, 1H), 7.95 (d, 1H, J=2.1 Hz), 8.20-8.10 (m, 1H), 8.01 (d, 1H,
J=8.7), 7.87 (s, 1H), 7.75 (d, 1H, J=6.6 Hz), 7.64 (dd, 1H, J=6.5,
2.7 Hz), 7.45 (d, 1H, J=8.7 Hz), 7.35-7.28 (m, 2H), 6.56 (d, 1H,
J=8.9 Hz), 5.60 (s, 1H), 3.85 (s, 3H), 2.99-2.80 (m, 12H); HRMS
(FAB): Calculated for C.sub.29H.sub.30F.sub.3N.sub.7O.sub.2
(M+H.sup.+): .delta. 566.2491, Found: 566.2511.
EXAMPLE G-11
4-Fluoro-3-[{5-(pyridin-3-yl)amino-2H-pyrazol-3-yl}methoxy]-N-[((4-pyrroli-
din-1-yl)-3-trifluoromethylphenyl)Benzamide
[0281] 158
[0282] Example G-11 was made in a similar manner to that described
for G-1, except that 4-(pyrrolidin-1-yl)-3-trifluoromethylaniline,
prepared according to the procedure described in WO 99/21845 (p.
58) for the preparation of 1-methyl-4-(4-nitrophenyl)piperazine,
was used in place of 4-isopropyl-3-methylaniline in step (d), and
N-(pyridin-3-yl)acetamide was used instead of
N-(diphenylmethyl)-N-methyl-4-(acetylamino)-benzenesu- lfonamide,
G-1b, in step (c): HPLC Rt=14.78 min; .sup.1H NMR (300 MHz,
Acetone-d.sub.6) .delta. 9.53 (br s, 1H), 8.55 (d, 1H, J=2.1H),
8.10 (d, 1H, J=2.7 Hz), 7.93-7.76 (m, 6H), 7.45-7.37 (m, 2H), 7.20
(d, 1H, J=9.0 Hz), 7.14-7.10 (m, 1H), 3.28-3.21 (m, 4H), 3.09-2.96
(m, 4H, 1.96-1.92 (m, 4H); HRMS (FAB): Calculated for
C.sub.28H.sub.27F.sub.3N.sub.6O (M+H.sup.+): 521.2277, Found:
521.266. Anal. calc'd for
C.sub.28H.sub.27F.sub.3N.sub.6O.0.9H.sub.2O: C, 62.65; H, 5.41; N,
15.66. Found C, 62.84; H, 5.33; N, 15.66.
EXAMPLE H-1
N-(4-Isopropyl-3-methyl-phenyl)-3-[2-(5-phenylamino-2-H-pyrazol-3-yl)-cycl-
opropyl]-benzamide
[0283] 159
[0284] Example H-1 was made in a similar manner to that described
for E-1, except that
N-(4-isopropyl-3-methylphenyl)-3-[2-(N-methoxy-N-methylcarbam-
oyl)cyclopropyl]benzamide, prepared as described below, was used
instead of in step (c): HPLC R.sub.t=16.47 min.; .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.13 (br s, 1H), 7.59-7.50 (m, 1H),
7.44-7.40 (m, 2H), 7.31-7.25 (m, 3H), 7.22-7.17 (m, 3H), 7.02 (d,
2H, J=7.8 Hz), 6.83 (t, 1H, J=7.5 Hz), 5.71 (s, 1H), 3.14-3.53 (m,
1H), 2.18-2.16 (m, 1H), 2.08-2.04 (m, 1H), 1.38-1.35 (m, 2H), 1.23
(d, 6H, J=6.9 Hz); HRMS (FAB): Calculated for
C.sub.29H.sub.30N.sub.4O (M+H.sup.+): 451.2498, Found: 451.2510.
Calculated for C.sub.29H.sub.30N.sub.4O.0.5H.sub.2O: C, 75.79; H,
6.80; N, 12.19. Found C, 75.83; H, 6.81; N, 12.19.
[0285]
N-(4-Isopropyl-3-methyl-phenyl)-3-[2-(N-methoxy-N-methyl-carbamoyl)-
-cyclopropyl]-benzamide was prepared as follows: To a solution of
trimethylsulfoxonium iodide (0.46 g, 2.01 mmol) in 10 mL of DMSO
was added sodium hydride (0.08 g, 2.01 mmol, 60% oil dispersion) at
-10.degree. C. After 30 min, to the reaction solution was added
dropwise a solution of 0.35 g (0.96 mmol) of
N-(4-isopropyl-3-methylphenyl)-3-[2-(-
N-methoxy-N-methylcarbamoyl)ethenyl]-benzamide, G-1e, in 5 ml of
DMSO. The reaction was allowed to warm to 25.degree. C. After 2 h,
the reaction was quenched with dropwise addition of 1N HCl. The
reaction mixture was partitioned between 30 mL dichloromethane and
30 mL of saturated NaHCO.sub.3 and the organic layer was dried over
sodium sulfate and evaporated to a give a crude yellow oil. The oil
was purified by silica gel chromatography using 75% hexane/25%
ethyl acetate to afford 0.11 g (29%) of
N-(4-isopropyl-3-methylphenyl)-3-[2-(N-methoxy-N-methylcarbamoyl-
)cyclopropyl]benzamide as a clear oil: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.18 (br s, 1H), 7.67-7.62 (m, 2H), 7.46-7.44
(m, 2 h), 7.36-7.26 (m, 3H), 7.20 (d, 1H, J=9.0 Hz), 3.67 (s, 3H),
3.21 (S, 3H), 2.55-2.45 (m, 2H), 2.31 (s, 3H), 1.65-1.59 (m, 1H),
1.37-1.31 (m, 1H), 1.21 (d, 6H, J=6.9 Hz); MS (ESI): Calculated for
C.sub.23H.sub.28N.sub.2O.sub.3 (M-H): 379, Found: 379.
EXAMPLE I-1
3-[({3-[(E)-2-(4-hydroxy-3-methoxyphenyl)ethenyl]-1H-pyrazol-5-yl}methyl)a-
mino]-N-(3-methyl-4-isopropylphenyl)Benzamide
[0286] 160
[0287] (a) Vanillin (9.12 g, 0.06 mole) was dissolved in 36 mL of
acetone and 12.5 mL of 50% aq. NaOH solution was added dropwise
with vigorous stirring. To the resulting solid was added 25 mL of
water and the dark red solution was refluxed for 5 min. This
reaction mixture was kept at room temperature for 24 h and
acidified with acetic acid. The reaction mixture was concentrated
to provide a yellow solid, which was filtered, washed with water
and dried to yield 10.2 g (88%) of
(E)-1-(3-methoxy-4-hydroxyphenyl)-1-butene-3-one, I-1a: .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 7.43 (d, 1H, J=16.3 Hz), 7.04-7.09
(m, 2H), 6.91 (d, 1H, J=7.9 Hz), 6.57 (d, 1H, J=16.3 Hz), 6.02 (s,
1H), 3.92 (s, 3H), 2.35 (s, 3H).
[0288] (b) Sodium metal (3.7 g, 0.16 mole) was dissolved in 100 mL
of absolute ethanol under an inert atmosphere. An solution of 16.2
g (953 mmol) of E-1-(3-methoxy-4-hydroxyphenyl)-1-butene-3-one,
I-1a, of ethanol was then added slowly over 30 min to the sodium
ethoxide solution. After the addition the reaction mixture was
stirred at for 15 min followed by the addition of diethyl oxalate
(7.7 g, 0.53 mole) and the dark red solution was stirred at room
temperature for 5 h. Concentrated HCl was added to the reaction
mixture until acidic. A dark yellow solid separated out which was
stirred in an ice bath for 1 h. The solid was filtered, washed and
dried to afford 8.4 g (55%) of the desired ethyl
(E)-6-(4-hydroxy-3-methoxyphenyl)-2,4-dioxo-5-hexenoate, I-1b:
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 14.86 (br s, 1H), 7.54
(d, 1H, J=15.9 Hz), 7.00 (d, 1H, J=8.3 Hz), 6.93 (s, 1H), 6.81 (d,
1H, J=8.3 Hz), 6.36-6.41 (m, 2H), 5.84 (s, 1H), 4.23 (q, 2H, J=7.2
Hz), 3.81 (s, 3H), 1.25 (t, 3H, J=7.2 Hz).
[0289] (c) To a solution of 8.1 g (27.7 mmol) of ethyl
6-(3'-methoxy-4'-hydroxyphenyl)-2,4-dioxo-5-hexenoate, I-1b, in 125
mL of acetic acid was added 1 mL of hydrazine (30.5 mmol) and the
reaction was stirred at 65.degree. C. or 90 min. The reaction was
cooled to room temperature and added slowly to 500 mL of ice cold
water upon which a white solid separated out. The solid was
filtered, washed and dried to afford 7.25 g (91%) of ethyl
(E)-3-[(2)-(4-hydroxy-3-methoxyphenyl)etheny-
l]-1H-pyrazole-5-carboxylate, I-1c: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 13.61 (s, 1H), 9.34 (s, 1H), 7.23 (d, 1H,
J=16.6 Hz), 7.19 (s, 1H), 6.92-7.01 (m, 2H), 6.84 (d, 1H, J=7.9
Hz), 4.32 (q, 2H, J=7.2 Hz) (s, 1H), 3.88 (s, 3H), 1.36 (t, 3H,
J=7.2 Hz); APCIMS m/z 289 [M+H].sup.+.
[0290] (d) To 40 ml of 1 M diisobutylauminum hydride in THF was
added dropwise 2.5 g (9.8 mmol) of ethyl
(E)-3-[2-(3-methoxy-4-hydroxyphenyl)et-
henyl]-1H-pyrazole-5-carboxylate, I-1c, in 25 mL of THF and the
reaction was stirred at room temperature. The reaction was
monitored by TLC and quenched after 6 h with water and extracted
with 3.times.150 mL of ethyl actate. The combined organic layers
were concentrated and the residue was purified using silica gel
column chromatography to afford 1.2 g (53%) of
4-{(E)-2-[5-(hydroxymethyl)-1H-pyrazol-3-yl]ethenyl}-2-methoxyphenol,
I-1d: .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 12.58 (br s, 1H),
9.13 (s, 1H), 7.12 (s, 1H), 6.98 (d, 1H, J=16.6 Hz), 6.88-6.92 (m,
2H), 6.74 (d, 1H, J=8.4 Hz), 6.34 (s, 1H), 5.10 (brs, 1H), 4.42 (d,
2H, J=5.6 Hz), 3.81 (s, 3H); APCIMS m/z 247 [M+H].sup.+.
[0291] (e) To 72 mg (0.3 mmol) of
4-{(E)-2-[5-(hydroxymethyl)-1H-pyrazol-3-
-yl]ethenyl}-2-methoxyphenol, I-1d, was added 1 mL of thionyl
chloride. After 10 min, the reaction was quenched by adding 10 mL
of ice cold water slowly and the mixture was extracted with
2.times.10 mL of ethyl acetate. The extracts were combined,
concentrated, and filtered through a silica gel plug. All the
washings were collected, the solvent removed under vacuo to obtain
32 mg (76%) of 4-{(E)-2-[5-(chloromethyl)-1H-pyrazol-3-yl-
]ethenyl}-2-methoxyphenol, I-1e: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 12.89 (brs, 1H), 9.20 (s, 1H), 7.11 (s, 1H),
7.04 (d, 1H, J=16.7 Hz), 6.74-6.90 (m, 2H), 6.75 (d, 1H, J=8.0 Hz),
6.45 (s, 1H), 4.69 (s, 2H), 3.81 (s, 3H); APCIMS m/z 265
[M+H].sup.+.
[0292] (f) To 132 mg (0.5 mmol) of
4-{(E)-2-[5-(chloromethyl)-1H-pyrazol-3-
-yl]ethenyl}-2-methoxyphenol, I-1e, in 2 mL of DMF was added
3-aminobenzoic acid (75 mg, 0.55 mmol) and an excess of NaHCO.sub.3
and the reaction mixture stirred at room temperature for 16 h. The
crude reaction mixture was filtered through a plug of silica gel to
remove NaHCO.sub.3 and washed with ethyl acetate. The filtrates
were collected and the solvents removed in vacuo to yield 60 mg of
crude
3-[({3-[(E)-2-(4-hydroxy-3-methoxyphenyl)ethenyl]-1H-pyrazol-5-yl}methyl)-
amino]benzoic acid, I-1f, which was redissolved in 2 mL of DMF. To
the resulting solution were added HATU (95 mg, 0.25 mmol) and
diisopropylethylamine (0.04 mL, 0.23 mmol) followed by
3-methyl-4-isopropylaniline (90 mg, 0.6 mmol) and the reaction
mixture stirred at room temperature for 16 h. After conventional
aqueous work-up, 16 mg of
3-[({3-[(E)-2-(4-hydroxy-3-methoxyphenyl)ethenyl]-1H-pyrazol-5-y-
l}methyl)amino]-N-(3-methyl-4-isopropylphenyl)benzamide, I-1, was
isolated using HPLC; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
9.91 (s, 1H), 7.51-7.54 (m, 2H), 7.03-7.20 (m, 7H), 6.98 (d, 1H,
J=15.5 Hz), 6.82-6.90 (m, 4H), 6.74 (d, 1H, J=9.0 Hz), 6.35 (s,
1H), 4.28 (s, 2H), 3.80 (s, 3H), 3.01-3.08 (m, 1H), 2.27 (s, 3H),
1.16 (d, 6H, J=6.0 Hz); APCIMS m/z 497 [M+H].sup.+.
EXAMPLE I-2
3-[({5-[(E)-2-(4-hydroxy-3-methoxyphenyl)ethenyl]-1H-pyrazol-3-yl}methyl)a-
mino]-N-phenylbenzamide
[0293] 161
[0294] Example I-2 was prepared in a similar manner to that
described for I-1, except that aniline was used in place of
3-methyl-4-isopropylaniline in step (f): HPLC R.sub.t=6.38 min.;
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.55 (d, 2H, J=9.0 Hz),
7.21-7.26 (m, 2H), 7.11-7.16 (m, 2H), 7.02-7.08 (m, 2H), 6.97-6.98
(m, 1H), 6.90 (d, 1H, J=15.0 Hz), 6.78-6.86 (m, 2H), 6.74 (d, 1H,
J=15.0 Hz), 6.65-6.67 (m, 1H), 6.33 (s, 1H), 4.27 (s, 2H), 3.77 (s,
3H); APCIMS m/z 441 [M+H].sup.+.
EXAMPLE J-1
4-Fluoro-N-[4-(imidazol-1-yl)-3-trifluoromethylphenyl]-3-[5-(6-methoxypyri-
din-3-yl)amino-2H-pyrazol-3-ylmethoxy]-benzamide
[0295] 162
[0296] (a) To a solution of 2-bromo-5-nitrobenzotrifluoride (1.50
g, 5.54 mmol) in toluene (11 mL) under argon purge was added
imidazole (0.57 g, 8.31 mmol), trans,trans-dibenzylidene acetone
(0.13 g, 0.56 mmol), 1,10-phenanthroline (1.00 g, 5.54 mmol),
cesium carbonate (1.99 g, 6.10 mmol) and
copper(II)triflate.cndot.benzene (0.015 g, 0.028 mmol). The slurry
was heated at 90.degree. .degree. C. for 18 h, and then cooled to
25.degree. C. The mixture was filtered through a silica gel plug
with 30 mL of ethyl acetate and the filtrate was concentrated. The
residue was purified by chromatography on silica gel (hexane/ethyl
acetate) to afford 0.83 g (58.2%) of
1-(4-Nitro-2trifluoromethylphenyl)-1H-imidazole, J-1a, as an amber
solid: HPLC Rt=10.45 min.; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 8.72 (d, 1H, J=2.1 Hz), 8.58 (dd, 1H, J=8.7, 2.7 Hz),
7.70-7.67 (m, 2H), 7.26 (s, 1H), 7.21 (s, 1H); MS (ESI): Calculated
for C.sub.10H.sub.6F.sub.3N.sub.3O.sub.2 (M+H.sup.+): 257, Found:
257.
[0297] (b) To a solution of 0.60 g (2.33 mmol) of
1-(4-nitro-2-trifluorome- thylphenyl)-1H-imidazole, J-1a, in 10 mL
of methanol was added 0.10 g of 10% Pd/C. The mixture was stirred
under a hydrogen atmosphere (1 atm) for 18 h, and then filtered
through a 0.22 .mu.M teflon filter membrane. The filtrate was
concentrated to afford 0.51 g (100%) of
1-(4-amino-2-trifluomethylphenyl)imidazole, J-1b, as a yellow
solid: HPLC Rt=8.89 min.; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
7.62 (s, 1H), 7.13-7.02 (m, 4H), 6.86 (d, 1H, J=8.4, 2.4 Hz). MS
(ESI): Calculated for C.sub.10H.sub.8F.sub.3N.sub.3 (M+H.sup.+):
228, Found: 228.
[0298] (c) To a solution of 3-acetoxy-4-fluorobenzoic acid (0.49 g,
2.48 mmol) in dichloromethane (5 mL) was added oxayl chloride (0.26
ml, 2.97 mmol) and DMF (0.1 mL). After 1 h, the mixture was
concentrated and then re-dissolved in 5 mL of dichloromethane. To
this solution was added 0.56 g (2.47 mmol) of
1-(4-amino-2-trifluomethylphenyl)imidazole, J-1b, and
diisopropylethylamine (0.39 mL, 2.48 mmol). After 1 h the solution
was partitioned between 30 mL of ethyl acetate and sat. aq. sodium
bicarbonate (2.times.20 mL). The organic layer was dried over
sodium sulfate and concentrated. The residue was purified by
chromatography on silica gel (1:1 hexane:ethyl acetate) to provide
0.49 g (57%) of
3-acetoxy-4-fluoro-N-[4-(imidazol-1-yl)-3-trifluoromethylphenyl]-benzamid-
e, J-1c, as clear oil: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.12-8.02 (m, 3H), 7.92-7.89 (m, 1H), 7.63 (s, 1H), 7.37-7.12 (m,
5H), 2.36 (s, 1H); MS (ESI): Calculated for
C.sub.19H.sub.13F.sub.4N.sub.3O.sub.3 (M+H.sup.+): 408, Found:
408.
[0299] (d) To a solution of 0.33 g (0.83 mmol)
3-acetoxy-4-fluoro-N-[4-(im-
idazol-1-yl)-3-trifluoromethylphenyl]-benzamide, J-1c, in 5 mL
acetone and 0.5 mL of methanol was added cesium carbonate (0.54 g,
1.65 mmol) and 2-chloro-N-methoxy-N-methyl-acetamide (0.15 g, 1.07
mmol) and the resulting mixture was stirred for 6 h at 45.degree.
C. After cooling to room temperature, the mixture was partitioned
between ethyl acetate and sat. brine (2.times.20 mL). The organic
layer was filtered though a silica gel plug and concentrated. The
residue was purified by titurated with diethyl ether (2.times.20
mL) to give 0.35 g (92%) of
4-fluoro-N-[4-(imidazol-1-yl)-3-trifluoromethylphenyl]-3-[(N-methoxy-N-me-
thylcarbamoyl)methoxy]benzamide, J-1d, as a white solid: Rt=11.95
min.; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.34 (d, 1H, J=2.4
Hz), 8.11 (dd, 1 h, J=8.7 Hz), 7.64-7.54 (m, 3H), 7.42 (d, 1H,
J=8.7 Hz), 7.22-7.15 (m, 2H), 6.97 (s, 1H), 502 (s, 2H), 3.72 (s,
3H), 3.06 (s, 3H); MS (ESI): Calculated for
C.sub.21H.sub.18F.sub.4N.sub.4O.sub.4 (M+H.sup.+): 467, Found:
467.
[0300] (e) To a -78.degree. C. solution of 0.13 g (0.71 mmol)
N-(6-methoxy-pyridin-3-yl)-thioacetamide in 5 mL of dry THF was
added dropwise 0.71 mL (1.42 mmol) of LDA (2.0 M in THF). The
solution was stirred for 15 min at -78.degree. C., warmed to
0.degree. C. for 1 h, and then cooled to -78.degree. C. To the
resulting solution was added, over a 5 min period, a solution of
0.15 g (0.32 mmol) 4fluoro-N-[4-(imidazol-1-y-
l)-3-trifluoromethyl-phenyl]-3-[(N-methoxy-N-methylcarbamoyl)methoxy]benza-
mide, J-1d, in 5 mL of THF. After stirring for 1 h at 0.degree. C.,
the reaction was quenched with 1 mL of 1:1 methanol:acetic acid.
The mixture was partitioned between ethyl acetate and sat. sodium
carbonate (2.times.20 mL) and the organic layer was dried over
sodium sulfate and concentrated. The residual yellow oil was
purified by radial chromatography (2 mm plate eluting with 1:2:1
hexane:ethyl acetate:dichloromethane) to give 0.091 g (49%) of
4-fluoro-N-[4-(imidazol-
-1-yl)-3-trifluoromethylphenyl]-3-[3-(6-methoxypyridin-3-yl)thiocarbamoyl--
2-oxo-propoxy]-benzamide, J-1e, as a yellow solid: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.74 (br s, 1H), 8.19 (d, 2H, J=2.7 Hz),
8.05 (d, 1H, J=2.7 Hz), 7.96 (dd, 2H, J=10.2, 3.0 Hz), 7.41 (dd,
1H, J=8.4, 2.7 Hz), 7.28 (s, 1H), 6.82-6.77 (m, 4H), 2.74 (s, 2H);
MS (ESI): Calculated for C.sub.27H.sub.21F.sub.4N.sub.5O.sub.4S
(M+H.sup.+): 588, Found: 588.
[0301] (e) To a solution of 0.061 g (0.10 mmol) of
4-fluoro-N-[4-(imidazol-
-1-yl)-3-trifluoromethyl-phenyl]-3-[3-(6-methoxypyridin-3-yl)thiocarbamoyl-
-2-oxo-propoxy]-benzamide, J-1e, in ethanol (2 mL) was added
hydrazine monohydrate (0.080 mL, 0.16 mmol) and acetic acid (0.09
mL, 0.16 mmol). After 2 h the solution was concentrated and the
residue was purified by chromatography on silica gel (1:2
hexane:ethyl acetate) to afford 0.031 g (54%) of
4-fluoro-N-(4-(imidazol-1-yl)-3-trifluoromethyl-phenyl)-3-[5-(6--
methoxy-pyridin-3-yl)amino-2H-pyrazol-3-ylmethoxy]benzamide, J-1,
as a off white solid: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.34 (s, 1H), 8.12-8.10 (m, 2H), 7.85 (t, 1H, 3.0 Hz), 7.69-7.57
(m, 2H), 7.41 (d, 1H, J=8.7 Hz), 7.22-7.13 (m, 2H), 6.97 (d, 1H,
J=6.0 Hz), 6.52 (d, 1H, J=9.0 Hz), 5.88 (s, 1H), 5.19 (s, 2H), 3.68
(s, 3H); LCESI: Calculated for
C.sub.27H.sub.21F.sub.4N.sub.7O.sub.3 (M+H.sup.+): 568, Found: 568.
Anal. calc'd for
C.sub.27H.sub.21F.sub.4N.sub.7O.sub.3.1.2CH.sub.2Cl.sub.2 Found: C,
50.59; H 3.52; N, 14.65. Found C, 50.69; H, 3.78; N, 14.79.
EXAMPLE J-2
4-Fluoro-3-[5-(6-methoxy-pyridin-3-yl)amino-2H-pyrazol-3-yl]methoxy-N-(4-p-
yrrolidin-1-yl-3-trifluoromethyl-phenyl)-benzamide
[0302] 163
[0303] Example J-2, was prepared in a similar manner to that
described for J-1, except that
4-(pyrrolidino)-3-trifluoromethylaniline (see example G-8) was used
in place of 1-(4-amino-2-trifluomethylphenyl)imidazole, J-1b, in
step (c): HPLC Rt=15.55 min.; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.98 (d, 1H, J=2.7 Hz), 7.85 (br s, 1H), 7.68-7.63 (m, 3H),
7.49 (dd, 1H, J=8.7, 3.0 Hz), 7.18-7.11 (m, 1H), 6.95 (d, 1H, J=8.7
Hz), 6.67 (d, 1H, J=8.7 Hz), 5.93 (s, 1H), 5.87 (s, 1H), 5.17 (s,
2H), 3.88 (s, 3H), 3.31-3.27 (m, 4H), 2.05-1.96 (m, 4H); HRMS
(MALDI): Anal. Calculated for C.sub.28H.sub.26F.sub.4N.sub.6O.sub.3
(M+Na.sup.+): 593.1900, Found: 593.1873. Anal. calc'd for
C.sub.28H.sub.26F.sub.4N.sub.6O.sub.3 C, 58.95; H, 4.95; N, 14.73.
Found C, 58.87; H, 4.91; N, 14.87.
EXAMPLE J-3
4-Fluoro-3-[5-(6-methoxypyridin-3-yl)amino-2H-pyrazol-3-yl]methoxy-N-(3-me-
thoxy-5-trifluoromethyl-phenyl)-benzamide
[0304] 164
[0305] Example J-3, was prepared in a similar manner to that
described for J-1, except that 5-methoxy-3-trifluoromethylaniline
(Aldrich) was used in place of
1-(4-amino-2-trifluomethylphenyl)imidazole, J-1b, in step (c):
Rt=14.50 min.; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.05 (d,
1H, J=3.0 Hz), 7.88 (s, 1H), 7.68 (dd, 1H, J=9.5, 2.1 Hz),
7.56-7.53 (m, 2H), 7.42-7.39 (m, 1H), 7.34 (s, 1H), 7.26-7.17 (m,
1H), 6.94 (s, 1H), 7.70 (d, 1H, J=9.0 Hz), 5.95 (s, 1H), 5.76 (br
s, 1H), 5.21 (s, 2H), 3.90 (s, 3H), 3.87 (s, 3H); HRMS (FAB):
Calculated for C.sub.25H.sub.21F.sub.4N.su- b.5O.sub.4
(M+Na.sup.+): 554.1427, Found: 554.1423. Anal. calc'd for
C.sub.25H.sub.21F.sub.4N.sub.5O.sub.4.0.2hexane: C, 57.35; H, 4.37;
N, 12.76. Found C, 57.00; H, 4.60; N, 13.00.
EXAMPLE K-1
N-(4-Isopropyl-3-methyl-phenyl)-3-(Isoquinolin-4-yl)methoxy-benzamide
[0306] 165
[0307] (a) A 2.6 M solution of n-BuLi in hexanes (7.4 mL, 19.2
mmol, 2.0 eq) was added to a solution of THF (40 mL) and ether (40
mL) and cooled to -78.degree. C. 4-Bromoisoquinoline (Aldrich, 2.0
g, 9.6 mmol, 1.0 eq) was added to the anion in one portion and the
dark orange solution was aged at -78.degree. C. for 30 minutes to
give a brown slurry. The mixture was treated with DMF (1.7 mL, 24.0
mmol, 2.5 eq) to give a brown solution. After 15 min, the reaction
was quenched with ethanol (40 mL). The resultant pale yellow
solution was treated with a saturated solution of ammonium chloride
(200 mL) and extracted with MTBE (3.times.200 mL). The combined
organic extracts were washed with brine (200 mL), dried over
magnesium sulfate, filtered and concentrated under reduced pressure
to give an orange solid (1.8 g). The crude product was purified by
flash chromatography over silica gel using 50% ethyl
acetate/cyclohexane (R.sub.f 0.5) to give
isoquinoline-4-carbaldehyde, K-1a, as a yellow solid (1.3 g, 84%):
HPLC R.sub.t=8.8 min.; TLC R.sub.f=0.2 (5% ethyl acetate/35%
cyclohexane/dichloromethane); .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 10.41 (s, 1H), 9.46 (s, 1H), 9.23 (d, 1H, J=8.5 Hz), 8.95
(s, 1H), 8.12 (d, 1H, J=8.1 Hz), 7.97 (t, 1H, J=7.6 Hz), 7.78 (t,
1H, J=7.3 Hz); MS (ESI) m/z 158 [M+H].sup.+.
[0308] (b) To a solution of 1.4 g (8.9 mmol) of
isoquinoline-4-carbaldehyd- e, K-1a, in ethanol (50 mL) at
0.degree. C. was added with sodium borohydride (372 mg, 9.8 mmol,
1.1 eq). The mixture was stirred at 0.degree. C. for 1.5 hours and
at room temperature for 1.0 h. The reaction was quenched with 25%
ammonium acetate (500 mL). The ethanol was removed under reduced
pressure and the resultant mixture was extracted with ethyl acetate
(3.times.500 mL). The combined organic extracts were washed with
brine (200 mL), dried over magnesium sulfate, filtered and
concentrated under reduced pressure to give a yellow oil (1.5 g).
The crude product was purified by radial chromatography over silica
gel using 5-10% methanol/dichloromethane to give, after
concentration from 50% ethyl acetate/cyclohexane,
4-(hydroxymethyl)isoquinoline, K-1b, as a tan solid (0.95 g, 68%):
HPLC R.sub.t=6.7 min.; TLC R.sub.f=0.2 (50% ethyl
acetate/cyclohexane); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
9.24 (s, 1H), 8.49 (s, 1H), 8.14 (d, 1H, J=8.4 Hz), 7.82 (t, 1H,
J=7.6 Hz), 7.70 (t, 1H, J=7.6 Hz), 5.36 (t, 1H, J=5.4 Hz), 4.95 (d,
2H, J=5.2 Hz); MS (FAB) m/z 160 [M+H].sup.+.
[0309] (c) A solution of 4-(hydroxymethyl)isoquinoline, K-1b, (900
mg, 5.7 mmol, 1.0 eq) in ethanol (10 mL) was treated with
concentrated hydrochloric acid (1.0 mL, 12.6 mmol, 2.1 eq). The
mixture was stirred for 30 min and then the solvent was removed
under reduced pressure. The tan residue was repeatedly evaporated
from toluene (3.times.10 mL) to give a tan solid (1.1 g, 100%). The
resulting unpurified 4-(hydroxymethyl)isoquinoline hydrochloride
was then treated with thionyl chloride (10 mL) and the resultant
mixture was heated to 70.degree. C. After 1.5 hours, the solvent
was removed under reduced pressure to give
4-(chloromethyl)isoquinoline hydrochloride, K-1c, as a tan solid
(1.1 g, 95%): HPLC R.sub.t=11.2 min.; .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 9.81 (s, 1H), 8.83 (s, 1H), 8.52 (d, 1H,
J=8.1 Hz), 8.42 (d, 1H, J=8.6 Hz), 8.23 (t, 1H, J=7.8 Hz), 8.01 (t,
1H, J=7.8 Hz), 5.41 (s, 2H); .sup.13C NMR (75 MHz, DMSO-d.sub.6)
.delta. 149.6, 136.2, 135.9, 134.1, 131.9, 131.4, 130.6, 127.8,
124.2; MS (FAB) m/z 177/179 [M.sup.-]. Anal. calc'd for
C.sub.10H,ClN.HCl: C, 56.10; H, 4.24; Cl, 33.12; N, 6.54. Found: C,
56.15; H, 4.32; Cl, 33.23; N, 6.37.
[0310] (d) A suspension of 3-methyl-4-isopropyl aniline
hydrochloride (6.7 g, 36.2 mmol, 1.0 eq) in THF (240 mL) was
treated with triethylamine (5.0 mL, 36.2 mmol, 1.0 eq). The gray
mixture was then treated with 3-hydroxybenzoic acid (Aldrich, 5.0
g, 36.2 mmol, 1.0 eq), followed by
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (7.6 g,
39.8 mmol, 1.1 eq). The resultant slurry was stirred at room
temperature for 18 h. The solvent was removed under reduced
pressure and the residue was extracted with water (200 mL) and
ethyl acetate (3.times.200 mL). The combined organic extracts were
washed with 5% KHSO.sub.4 (2.times.200 mL), water, (200 mL), brine
(200 mL), dried over magnesium sulfate, filtered and concentrated
under reduced pressure to give a brown solid (8.0 g). The crude
product was purified by flash chromatography on silica gel using
3-5% methanol/dichloromethane to give 3-hydroxy-N-(4-isopropyl--
3-methyl-phenyl)-benzamide, K-1d, as an off-white solid (1.1 g,
11%): HPLC R.sup.t=13.9 min.; TLC R.sub.f=0.4 (3%
methanol/dichloromethane); .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. 9.99 (s, 1H), 9.71 (s, 1H), 7.55-7.29 (m, 5H), 7.19 (d, 1H,
J=8.1 Hz), 7.97-7.95 (m, 1H), 3.09-3.06 (m, 1H), 2.29 (s, 3H), 1.18
(d, 6H, J=6.9 Hz); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
165.6, 157.7, 142.0, 136.9, 134.9, 129.7, 125.0, 122.4, 118.8,
118.7, 118.4, 114.8, 28.7, 23.5, 19.5; MS (ESI) m/z 268
[M-H].sup.-. Anal. calc'd for C.sub.17H.sub.19NO.sub.2: C, 75.81;
H, 7.11; N, 5.20. Found: C, 75.35; H, 7.23; N, 5.12.
[0311] (e) To a clear solution of
3-hydroxy-N-(4-isopropyl-3-methyl-phenyl- )-benzamide, K-1d, (0.538
g, 2.0 mmol, 1.0 eq) in acetone (40 mL) was added cesium carbonate
(5.2 g, 16.0 mmol, 8.0 eq). The resultant mixture was stirred at
room temperature for 30 minutes, treated with
4-chloromethyl-isoquinoline hydrochloride, K-1c, (0.469 g, 2.2
mmol, 1.1 eq) and warmed to 50.degree. C. After 18 hours, the
reaction was diluted with water (200 mL) and extracted with ethyl
acetate (3.times.200 mL). The combined organic extracts were washed
with brine (300 mL), dried over magnesium sulfate, filtered and
concentrated under reduced pressure to give a yellow oil (1.7 g).
The oil was purified by radial chromatography over silica gel
eluting with 50% ethyl acetate/cyclohexane (R.sub.f 0.3) to give,
after concentration from MTBE, N-(4-isopropyl-3-methyl-phenyl)-3-
-(Isoquinolin-4-yl)methoxy-benzamide, K-1, (0.69 g, 83%) as a light
yellow foam: HPLC R.sub.t=16.9 min.; TLC R.sub.f=0.6 (3%
methanol/chloroform); .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
10.06 (s, 1H), 9.37 (s, 1H), 8.69 (s, 1H), 8.22 (d, 1H, J=8.1 Hz),
8.17 (d, 1H, J=8.5 Hz), 7.90 (t, 1H1J=7.7 Hz), 7.77 (t, 1H, J=7.5
Hz), 7.69 (s, 1H), 7.60-7.34 (m, 5H), 7.21 (d, 1H, J=8.4 Hz), 5.66
(s, 2H), 3.09-3.07 (m, 1H), 2.30 (s, 3H), 1.18 (d, 6H, J=6.8 Hz);
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. 165.1, 158.6, 153.9,
143.5, 142.2, 136.8, 136.7, 135.0, 134.1, 131.5, 130.0, 128.6,
128.3, 128.0, 126.0, 125.0, 123.6, 122.5, 120.7, 118.8, 118.3,
114.3, 66.2, 28.7, 23.5, 19.4; MS (ESI) m/z 411 [M+H].sup.+. Anal.
calc'd for C.sub.27H.sub.26N.sub.2O.sub.2.0.3H120: C, 77.97; H,
6.45; N, 6.74. Found: C, 77.76; H, 6.64; N, 6.48.
EXAMPLE K-2
3-(Isoquinolin-4-yl)methoxy-N-(3,4,5-trimethoxyphenyl)Benzamide
Hydrochloride
[0312] 166
[0313] Example K-2 was prepared in a similar manner to that
described for K-1, except that 3,4,5-trimethoxyaniline (Aldrich)
was used in place of 3-methyl-4-isopropylaniline in step (d). The
product was isolated as the hydrochloride salt as follows. A
solution of 3-(isoquinolin-4-yl)methoxy--
N-(3,4,5-trimethoxyphenyl)benzamide (116 mg, 0.26 mmol) in ethanol
was treated with concentrated hydrochloric acid (0.1 mL, 1.2 mmol).
After several minutes, the solvent was removed under reduced
pressure to give
3-(isoquinolin-4-yl)methoxy-N-(3,4,5-trimethoxyphenyl)benzamide
hydrochloride as a white solid (126 mg, 100%): HPLC R.sub.t=13.4
min.; TLC R.sub.f=0.5 (5% methanol/chloroform); .sup.1H NMR (300
MHz, DMSO-d.sub.6) .delta. 10.10 (s, 1H), 9.70 (s, 1H), 8.72 (s,
1H), 8.41 (d, 1H, J=8.1 Hz), 8.30 (d, 1H, J=8.5 Hz), 8.10 (t, 1H,
J=7.8 Hz), 7.90 (t, 1H, J=7.6 Hz), 7.67 (s, 1H), 7.53 (d, 1H, J=7.7
Hz), 7.41 (t, 1H, J=7.9 Hz), 7.29 (dd, 1H, J=8.2, 1.6 Hz), 7.16 (s,
2H), 5.70 (s, 2H), 3.66 (s, 6H), 3.53 (s, 3H); .sup.13 C NMR (75
MHz, DMSO-d.sub.6) .delta. 165.1, 158.2, 152.9, 149.5, 136.7,
136.1, 135.8, 135.6, 134.2, 133.5, 131.0, 130.9, 130.3, 130.0,
127.6, 124.4, 121.0, 118.6, 114.5, 98.6, 65.4, 60.5, 56.1; MS (ESI)
m/z 445 [M+H].sup.+. Anal. calc'd for
C.sub.25H.sub.22N.sub.4O.sub.3.HCl.0.5H.sub.2O: C, 63.73; H, 5.35;
N, 5.72. Found: C, 63.52; H, 5.51; N, 5.50.
EXAMPLE K-3
3-(Isoquinolin-4-yl)methoxy-N-(2-methyl-quinolin-6-yl)-benzamide
Hydrochloride
[0314] 167
[0315] Example K-3, which was isolated as a dihydrochloride salt as
described in Example K-2, was prepared in a similar manner to that
described for K-1, except that 6-amino-2-methylquinoline
(Maybridge) was used in place of 3-methyl-4-isopropylaniline in
step (d): HPLC R.sub.t=13.2 min.; TLC R.sub.f=0.5 (5%
methanol/chloroform); .sup.1H NMR (300 MHz, DMSO-d.sub.6 w/
D.sub.2O) .delta. 9.78 (s, 1H), 9.0 (d, 1H, J=8.7 Hz), 8.84 (d, 2H,
J=1.9 Hz), 8.55 (d, 1H, J=8.2 Hz), 8.45-8.40 (m, 2H) 8.27-8.22 (m,
2H), 8.05 (t, 1H, J=7.5 Hz), 7.92 (d, 1H, J=8.7 Hz), 7.83 (t, 1H,
J=1.8 Hz), 7.72 (d, 1H, J=7.7 Hz), 7.60 (t, 1H, J=8.0 Hz), 7.48
(dd, 1H, J=8.2, 1.8 Hz), 5.84 (s, 2H), 2.95 (s, 3H); .sup.13C NMR
(75 MHz, DMSO-d.sub.6 w/ D.sub.2O) .delta. 167.9, 159.8, 158.2,
151.2, 146.7, 140.7, 137.6, 137.4, 136.3, 135.4, 132.6, 132.2,
132.1, 132.0, 130.3, 129.2, 129.1, 125.9, 125.7, 122.9, 122.7,
120.8, 118.7, 116.1, 66.9, 22.0; MS (FAB) m/z 420 [M+H].sup.+.
Anal. calc'd for C.sub.27H.sub.21N.sub.3O.sub.2.2HCl. 0.3H.sub.2O:
C, 65.14; H, 4.78; N, 8.44. Found: C, 65.18; H, 4.84; N, 8.38.
EXAMPLE K-4
3-(Isoquinolin-4-yl)methoxy-N-(2-methyl-4-methylsulfanyl-quinolin-6-yl)-be-
nzamide Hydrochloride
[0316] 168
[0317] Example K-4, which was isolated as a dihydrochloride salt as
described in Example K-2, was prepared in a similar manner to that
described for K-1, except that
2-methyl-4-methylsulfanyl-quinolin-6-ylami- ne, K-4d (vide infra),
was used in place of 3-methyl-4-isopropylaniline in step (d): HPLC
R.sub.t=14.4 min.; TLC R.sub.f=0.5 (5% methanol/chloroform);
.sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 11.10 (s, 1H), 9.80 (s,
1H), 9.07 (s, 1H), 8.89 (s, 1H), 8.54 (t, 2H, J=7.6 Hz), 8.46 (d,
1H, J=8.5 Hz), 8.33 (d, 1H, J=9.2 Hz), 8.22 (t, 1H, J=7.5 Hz), 8.04
(t, 2H, J=7.6 Hz), 7.95 (s, 1H), 7.79-7.78 (m, 2H), 7.63 (t, 1H,
J=7.9 Hz), 7.52 (d, 1H, J=6.6 Hz), 5.88 (s, 2H), 2.94 (s, 3H), 2.92
(s, 3H); HRMS (FAB) calcd for C.sub.28H.sub.23N.sub.3O.sub.2S
[M+H].sup.+ 466.1589, found 466.1577.
[0318] The intermediate
2-methyl-4-methylsulfanyl-quinolin-6-ylamine, K-4d, was prepared as
follows: 169
[0319] (a) A solution of 2-methyl-quinolin-4-ol (Aldrich, 9.2 g,
57.9 mmol. 1.0 eq) in concentrated sulfuric acid (60 mL) was cooled
to 0.degree. C. and treated with fuming nitric acid (3.9 mL, 57.9
mmol, 1.0 eq). The dark orange solution was stirred at 0.degree. C.
for 15 minutes and then poured into ice water (1000 mL) to give a
yellow precipitate. After standing for 18 hours, the mixture was
filtered and the yellow precipitate was washed with ice water to
give 2-methyl-6-nitro-quinolin-4- -ol, K-4a, as a yellow solid (6.9
g, 58%): HPLC R.sub.t=6.8 min.; .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. 8.10 (s, 1H), 8.42 (dd, 1H, J=9.2, 2.6 Hz), 7.70 (d, 1H,
J=9.1 Hz), 6.15 (s, 1H), 2.42 (s, 3H); MS (ESI) m/z 203
[M-H].sup.-.
[0320] (b) A solution of 2-methyl-6-nitro-quinolin-4-ol, K-4a, (6.9
g, 33.8 mmol) in phosphorus oxychloride (70 mL) was heated to
95.degree. C. After 1 hour, the reaction mixture gives no starting
material by HPLC analysis. The black solution was cooled to room
temperature and poured into ice water (500 mL). The aqueous layer
was made basic by the addition of concentrated ammonium hydroxide,
and extracted with chloroform (3.times.500 mL). The combined
organic extracts were washed with 5.0 N ammonium hydroxide (500
mL), water (500 mL), brine (500 mL), dried over magnesium sulfate,
filtered and concentrated under reduced pressure. The crude product
was passed through a silica gel plug, eluting with 20% ethyl
acetate/chloroform, to give a brown solid (6.4 g). The product was
crystallized from hot ethanol to give
4-chloro-2-methyl-6-nitro-quinoline- , K-4b, as tan needles (3.6 g,
48%): mp 142-144.degree. C.; HPLC R.sub.t=13.5 min.; TLC
R.sub.f=0.3 (20% ethyl acetate/cyclohexane); .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. 8.92, (d, 1H, J=2.5 Hz), 8.52 (dd, 1H, J=9.2,
2.6 Hz), 8.20 (d, 1H, J=9.2 Hz), 7.94 (s, 1H), 2.73 (s, 3H); MS
(ESI) m/z 223 [M+H].sup.+.
[0321] (c) A solution of 4-chloro-2-methyl-6-nitro-quinoline, K-4b,
(372 mg, 1.7 mmol, 1.0 eq) in anhydrous ethanol (70 mL) was treated
with sodium thiomethoxide (619 mg, 8.8 mmol, 5.2 eq), and the
resultant green slurry was heated at 50.degree. C. After 2.5 hours,
the solvent was removed under reduced pressure and the residue was
partitioned between water (100 mL) and chloroflorm,(3.times.100
mL). The combined organic extracts were washed with brine (100 mL),
dried over magnesium sulfate, filtered and concentrated under
reduced pressure to give a yellow solid (430 mg). The crude product
was purified by radial chromatography over silica gel using 5%
ethyl acetate/chloroform, followed by crystallization from hot
ethanol, to give 2-methyl-4-methylsulfanyl-6-nitro-quinoline, K-4c,
as yellow needles (250 mg, 46%): HPLC R.sub.t=13.1 min.; TLC
R.sub.f=0.5 (10% ethyl acetate/chloroform); .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. 8.70 (d, 1H, J=2.4 Hz), 8.32 (dd, 1H, J=9.2,
2.5 Hz), 7.97 (d, 1H, J=9.2 Hz), 7.35 (s, 1H), 2.61 (s, 3H), 2.58
(s, 3H); MS (ESI) m/z 235 [M+H].sup.+.
[0322] (d) A mixture of
2-methyl-4-methylsulfanyl-6-nitro-quinoline, K-4c, (200 mg, 0.85
mmol, 1.0 eq), and tin(II) chloride dihydrate (965 mg, 4.27 mmol,
5.0 eq) in anhydrous ethanol (50 mL) was heated to 75.degree. C. to
give an orange solution. After 30 minutes, the reaction mixture was
concentrated under reduced pressure to about 10 mL. The resultant
mixture was poured into ice water (100 mL), and the aqueous layer
was adjusted to pH 10 using concentrated ammonium hydroxide. The
aqueous layer was extracted with ethyl acetate (3.times.100 mL),
the combined organic layers were washed with brine (100 mL), dried
over magnesium sulfate, filtered and concentrated under reduced
pressure to give a yellow solid (250 mg). The crude product was
purified by radial chromatography over silica gel using 30% ethyl
acetate/dichloromethane w/ 0.5% methanol to give
2-methyl-4-methylsulfanyl-quinolin-6-ylamine, K-4d, as a yellow
solid (70 mg, 40%): HPLC R.sub.t=9.0 min.; TLC R.sub.f=0.6 (5%
methanol/chloroform); .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
7.58 (d, 1H, J=8.9 Hz), 7.08 (dd, 1H, J=8.9, 2.4 Hz), 7.02 (s, 1H),
6.90 (d, 1H, J=2.4 Hz), 5.58 (s, 2H), 2.58 (s, 3H), 2.51 (s, 3H);
MS (ESI) m/z 205 [M+H].sup.+.
EXAMPLE K-5
3-(Pyridin-3-yl)methoxy-N-(3,4,5-trimethoxyphenyl)Benzamide
[0323] 170
[0324] Example K-5, which was obtained as a white solid in a 62%
yield, was prepared in a similar manner to that described for K-1,
except that 3,4,5-trimethoxyaniline was used in place of
3-methyl-4-isopropylaniline in step (d), and 3-picolyl chloride
hydrochloride was used in place of 4-(chloromethyl)isoquinoline
hydrochloride, K-1c, in step (e): HPLC R.sub.t=18.1 min.; TLC
R.sub.f=0.5 (5% methanol/chloroform); .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.73 (s, 1H), 8.60 (d, 1H, J=2.9 Hz), 8.22 (s,
1H), 7.91 (d, 1H, J=7.4 Hz), 7.61 (s, 1H), 7.52-7.41 (m, 3H), 7.16
(d, 1H, J=7.6 Hz), 7.04 (s, 2H), 5.19 (s, 2H), 3.87 (s, 6H), 3.85
(s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. 165.2, 158.5,
153.3, 147.9, 147.4, 136.6, 136.4, 134.9, 134.0, 133.0, 130.0,
124.0, 119.7, 118.8, 113.3, 98.0, 67.4, 61.0, 56.1; MS (FAB) m/z
395 [M+H].sup.+. Anal. calc'd for C.sub.22H.sub.22N.sub.2O.sub.5:
C, 66.99; H, 5.62; N, 7.10. Found: C, 67.00; H, 5.65; N, 7.09.
EXAMPLE K-6
N-(Naphthalen-2-yl)-3-(pyridin-3-yl)Methoxybenzamide
[0325] 171
[0326] Example K-6, which was obtained as a white solid in a 19%
yield, was prepared in a similar manner to that described for K-1,
except 2-aminonaphthalene (Aldrich) was used in place of
3-methyl-4-isopropylani- line in step (d), and 3-picolyl chloride
hydrochloride was used in place of 4-(chloromethyl)isoquinoline
hydrochloride, K-1c, in step (e): HPLC R.sub.t=22.5 min.; TLC
R.sub.f=0.4 (5% methanol/chloroform); .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 10.41 (s, 1H), 8.72 (d, 1H, J=1.8 Hz), 8.58
(dd, 1H, J=4.8, 1.6 Hz), 8.44 (d, 1H, J=1.5 Hz), 7.94-7.81 (m, 5H),
7.68-7.62 (m, 2H), 7.53-5.28 (m, 5H), 5.27 (s, 2H); .sup.13C NMR
(125 MHz, DMSO-d.sub.6) .delta. 165.6, 158.5, 149.6, 149.5, 137.0,
136.7, 136.1, 133.7, 132.8, 130.4, 130.0, 128.5, 127.8, 127.7,
126.7, 125.2, 124.0, 121.3, 120.7, 118.4, 117.0, 114.4, 67.6; MS
(FAB) m/z 355 [M+H].sup.+. Anal. calcd for
C.sub.23H.sub.18N.sub.2O.sub.2: C, 77.95; H, 5.12; N, 7.70. Found:
C, 77.41; H, 5.22; N, 7.79.
EXAMPLE K-7
N-(1-Allyl-1H-indol-5-yl)-3-(pyridin-3-yl)methoxy-benzamide
[0327] 172
[0328] Example K-7, which was obtained as a light yellow solid in a
25% yield, was prepared in a similar manner to that described for
K-1, except 1-allyl-1H-indol-5-ylamine, K-7b (vide infra), was used
in place of 3-methyl-4-isopropylaniline in step (d), and 3-picolyl
chloride hydrochloride was used in place of
4-(chloromethyl)isoquinoline hydrochloride, K-1c, in step (e): HPLC
R.sub.t=21.6 min.; TLC R.sub.f=0.7 (5% methanol/chloroform);
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.14 (s, 1H), 8.77 (d,
1H, J=1.5 Hz), 8.62 (d, 1H, J=5.1 Hz), 8.05 (s, 1H), 7.98 (d, 1H,
J=8.1 Hz), 7.68-7.66 (m, 2H), 7.53-7.28 (m, 7H), 6.49 (d, 1H, J=2.9
Hz), 6.10-6.01 (m, 1H), 5.30 (s, 2H), 5.20 (d, 1H, J=10.3 Hz), 5.05
(d, 1H, J=17.3 Hz), 4.86 (d, 2H, J=5.2 Hz); HRMS (FAB) calcd for
C.sub.24H.sub.21N.sub.3O.sub.2 [M+H].sup.+ 384.1712, found
384.1708. The intermediate 1-allyl-1H-indol-5-ylamine, K-7b, was
prepared as follows: 173
[0329] (a) A solution of 5-nitro-1H-indole (Acros, 2.0 g, 13.5
mmol, 1.0 eq) in DMF (125 mL) was cooled to 0.degree. C., treated
with sodium hydride (60% in mineral oil, 600 mg, 14.9 mmol, 1.1 eq)
and stirred for 2.0 hours. The resulting red solution was treated
with allyl bromide (1.3 mL, 14.9 mmol, 1.1 eq) and stirred for 1.0
hour at room temperature. The reaction was diluted with water (1.0
L) and extracted with MTBE (3.times.500 mL). The combined organic
extracts were washed with water (500 mL), brine (200 mL), dried
over magnesium sulfate, filtered and concentrated under reduced
pressure to give a brown oil (2.7 g). The crude product was
purified by radial chromatography over silica gel using 5-30% ethyl
acetate/cyclohexane to give 1-allyl-5-nitro-1H-indole, K-7a, as a
light yellow oil (2.5 g, 90%): TLC R.sub.f=0.3 (5% ethyl
acetate/cyclohexane); .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
6.60 (d, J=2.2 Hz, 1H), 8.11 (dd, 1H, J=9.2, 2.2 Hz), 7.34 (d, 1H,
J=9.2 Hz), 7.26-7.24 (m, 1H), 6.70 (d, 1H, J=3.3 Hz), 6.06-5.94 (m,
1H), 5.26 (dd, 2H, J=10.3, 0.7 Hz), 5.08 (dd, 1H, J=17.3, 0.7 Hz),
4.79 (d, 2H, J=5.5 Hz); MS (FAB) m/z 203 [M+H].sup.+.
[0330] (b) A mixture of water (20 mL), methanol (20 mL), zinc (4.2
g, 64.2 mmol, 5.6 eq) and 1-allyl-5-nitro-1H-indole, K-7a, (2.3 g,
11.4 mmol, 1.0 eq) was treated with concentrated sulfuric acid (6
mL, 333 mmol, 29 eq) at 0.degree. C. The mixture was gradually
warmed to room temperature. After 18 hours, the mixture was
filtered, and the pH of the resulting brown solution was adjusted
to 9 with a saturated sodium bicarbonate solution. The resulting
slurry was filtered and extracted with chloroform (3.times.500 mL).
The combined organic extracts were washed with brine (200 mL),
dried over magnesium sulfate, filtered and concentrated under
reduced pressure to give a black oil (0.94 g). The crude product
was purified by radial chromatography over silica gel using 40%
ethyl acetate/cyclohexane with 0.5% methanol to give
1-allyl-1H-indol-5-ylamine- , K-7b, as a dark brown oil (233 mg,
5%): TLC R.sub.f=0.3 (30% ethyl acetate/cyclohexane); .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 7.16 (d, 1H, J=8.4 Hz), 7.07-7.03 (m,
2H), 6.77 (dd, 1H, J=8.6, 2.0 Hz), 6.36 (d, 1H, J=2.9 Hz),
6.04-5.91 (m, 1H), 5.18 (dd, 1H, J=10.3, 1.1 Hz), 5.05 (dd, 1H,
J=16.9, 1.1 Hz), 4.67 (d, 2H, J=5.5 Hz), 3.8 (br s, 2H); MS (FAB)
m/z 171 [M-H].sup.-.
EXAMPLE K-8
3-(Pyridin-3-yl)methoxy-N-quinolin-6-yl-benzamide
[0331] 174
[0332] Example K-8, which was obtained as a white solid in a 20%
yield, was prepared in a similar manner to that described for K-1,
except 6-aminoquinoline (Fluka) was used in place of
3-methyl-4-isopropylaniline in step (d), and 3-picolyl chloride
hydrochloride was used in place of 4-(chloromethyl)isoquinoline
hydrochloride, K-1c, in step (e): HPLC R.sub.t=17.0 min.; TLC
R.sub.f=0.6 (5% methanol/chloroform); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.60 (s, 1H), 8.86 (d, 1H, J=3.3 Hz), 8.78
(s, 1H), 8.63-8.60 (m, 2H), 8.39 (d, 1H, J=8.5 Hz), 8.12-8.04 (m,
2H), 7.98 (d, 1H, J=8.1 Hz), 7.74-7.70 (m, 2H), 7.67-7.49 (m, 3H),
7.37-7.34 (m, 1H), 5.32 (s, 2H); HRMS (FAB) calcd for
C.sub.22H.sub.17N.sub.3O.sub.2 [M+H].sup.+ 356.1399, found
384.1406.
EXAMPLE K-9
N-(2-Methyl-quinolin-6-yl)-3-(pyridin-3-yl)methoxy-benzamide
[0333] 175
[0334] Example K-9, which was obtained as a white solid in a 16%
yield, was prepared in a similar manner to that described for K-1,
except 6-amino-2-methylquinoline (Maybridge) was used in place of
3-methyl-4-isopropylaniline in step (d), and 3-picolyl chloride
hydrochloride was used in place of 4-(chloromethyl)isoquinoline
hydrochloride, K-1c, in step (e): HPLC R.sub.t=19.9 min.; TLC
R.sub.f=0.2 (3% methanol/chloroform); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.54 (s, 1H), 8.77 (d, 1H, J=0.7 Hz), 8.62
(dd, 1H, J=4.8, 1.4 Hz), 8.52 (d, 1H, J=1.9 Hz), 8.26 (d, 1H, J=8.8
Hz), 8.05-7.93 (m, 3H), 7.71-7.69 (m, 2H), 7.66-7.43 (m, 3H),
7.36-7.33 (m, 1H), 5.32 (s, 2H), 2.69 (s, 3H); .sup.13C NMR (125
MHz, DMSO-d.sub.6) .delta. 165.4, 158.1, 157.4, 149.3, 149.2,
144.5, 136.3, 135.8, 132.4, 129.7, 128.5, 126.3, 124.1, 123.6,
122.4, 120.4, 118.1, 116.4, 114.1, 67.3, 24.7; HRMS (FAB) calcd for
C.sub.23H.sub.19N.sub.3O.sub.2 [M+H].sup.+ 370.1556, found
370.1549.
EXAMPLE K-10
N-(4-Isopropyl-3-methyl-phenyl)-4-fluoro-3-(Isoquinolin-4-yl)methoxy-benza-
mide
[0335] 176
[0336] Example K-10 was prepared in a similar manner to that
described for K-1, except 4-fluoro-3-hydroxybenzoic acid was used
in place of 3-hydroxybenzoic acid in step (d): .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 9.28 (s, 1H), 8.63 (s, 1H), 8.11 (d, 1H,
J=8.23 Hz) 8.05 (d, 1H, J=8.11 Hz), 7.79 (m, 2H), 7.68 (m, 2H),
7.40 (m, 3H), 7.22 (m, 2H), 5.60 (s, 2H), 3.12 (br, 1H), 2.35 (s,
3H), 1.22 (d, 6H, J=6.9 Hz). MS (FAB) m/z 429 [M+H].sup.+. Anal.
calcd for C.sub.27H.sub.25FN.sub.2O.sub.2. 0.8H.sub.2O: C, 73.22;
H, 6.05; N, 6.33. Found: C, 73.21; H, 5.73; N, 6.19.
EXAMPLE K-11
N-(4-Isopropyl-3-methyl-phenyl)-4-methyl-3-(Isoquinolin-4-yl)methoxy-benza-
mide
[0337] 177
[0338] Example K-11 was prepared in a similar manner to that
described for K-1, except 4-methyl-3-hydroxybenzoic acid was used
in place of 3-hydroxybenzoic acid in step (d): .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 9.29 (s, 1H), 8.67 (s, 1H), 8.07 (s, 1H,)
8.04 (s, 1H), 7.80-7.64 (m, 4H), 7.46-7.30 (m, 4H), 7.23 (m, 1H),
5.54 (s, 2H), 3.19-3.06 (m, 1H), 2.36 (s, 3H), 2.25 (s, 3H), 1.23
(d, 6H, J=6.84 Hz). MS (ESI) m/z 425 [M+H].sup.+. Anal. calc'd for
C.sub.28H.sub.28N.sub.2O.sub.2: C, 79.22; H, 6.65; N, 6.60. Found:
C, 79.27; H, 6.74; N, 6.60.
EXAMPLE K-12
N-(4-Isopropyl-3-methyl-phenyl)-4-chloro-3-(Isoquinolin-4-yl)methoxy-benza-
mide
[0339] 178
[0340] Example K-12 was prepared in a similar manner to that
described for K-1, except 4-chloro-3-hydroxybenzoic acid was used
in place of 3-hydroxybenzoic acid in step (d): .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 9.30 (br, s, 1H), 8.70 (br, s, 1H),
8.14-8.04 (m, 2H), 7.82-7.65 (m, 4H), 7.50-7.32 (m, 4H), 7.23 (m,
1H), 5.61 (s, 2H), 3.13 (m, 1H), 2.36 (s, 3H), 1.23 (d, 6H, J=6.89
Hz). EIMS m/z 444 [M.sup.+]. Anal. calc'd for
C.sub.27H.sub.25ClN.sub.2O.sub.2: C, 72.88; H, 5.66; N, 6.30.
Found: C, 72.86; H, 5.71; N, 6.24.
EXAMPLE L-1
3-(6-Aminopyridin-3-yl)methoxy-N-(4-Isopropyl-3-methyl-phenyl)-benzamide
[0341] 179
[0342] (a) A solution of 6-chloronicotinic acid (Aldrich, 10.0 g,
63.7 mmol, 1.0 eq) in dioxane (300 mL) was treated with a 1.0 M
solution of borane in THF (320 mL, 329 mmol, 5.0 eq) at room
temperature. The resultant orange solution was stirred for one hour
and then heated to 75.degree. C. After 2.5 h, the reaction was
quenched with ethanol (100 mL), and the solvent was removed under
reduced pressure. The crude product was stirred in 1.2 M HCl (350
mL) and the pH was subsequently adjusted to 9 with solid NaOH. The
aqueous layer was extracted with ethyl acetate (3.times.200 mL) and
the combined organic extracts were washed with water (200 mL),
brine (200 mL), dried over magnesium sulfate, filtered and
concentrated under reduced pressure to give a yellow oil (23.4 g).
The crude product was purified by flash chromatography over silica
gel using 30-60% ethyl acetate/cyclohexane to give
(6-chloropyridin-3-yl)methanol, L-1a, as a white crystalline solid
(5.0 g, 54%): HPLC R.sub.t=2.9 min.; TLC R.sub.f=0.5 (5%
methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 8.35 (d, 1H, J=1.9 Hz), 7.79 (dd, 1H, J=8.2, 2.4 Hz), 7.48
(d, 1H, J=8.2 Hz), 5.42 (t, 1H, J=5.6 Hz), 4.53 (d, 2H, J=5.7 Hz);
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. 148.9, 148.4, 138.5,
137.6, 124.2, 60.2; MS (ESI) m/z 144 [M+H].sup.+.
[0343] (b) A solution of the (6-chloropyridin-3-yl)-methanol, L-1a,
(186 mg, 1.3 mmol, 1.0 eq),
3-hydroxy-N-(4-isopropyl-3-methyl-phenyl)-benzamid- e, K-1d, (350
mg, 1.3 mmol, 1.0 eq) and triphenylphosphine (1.0 g, 3.9 mmol, 3.0
eq) in THF (15 mL) was protected from light and treated with
diethyl azodicarboxylate (0.62 mL, 3.9 mmol, 3.0 eq). After 18 h,
the resultant light yellow solution was poured into 50% brine (200
mL) and extracted with ethyl acetate (3.times.100 mL). The combined
organic extracts were washed with brine (200 mL), dried over
magnesium sulfate, filtered through a silica gel plug, and
concentrated under reduced pressure to give a yellow oil (2.6 g).
The crude product was purified by radial chromatography over silica
gel using 5-10% ethyl acetate/45% cyclohexane/dichloromethane to
give 3-(6-chloropylidin-3-yl)methoxy-N-(4--
isopropyl-3-methyl-phenyl)-benzamide, L-1b, as a white solid (380
mg, 74%): HPLC R.sub.t=16.2 min.; TLC R.sub.f=0.4 (30% ethyl
acetate/cyclohexane); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
10.07 (s, 1H), 8.56 (d, 1H, J=2.3 Hz), 7.99 (dd, 1H, J=8.2, 2.4
Hz), 7.60-7.44 (m, 6H), 7.27-7.19 (m, 2H), 5.25 (s, 2H), 3.31-3.03
(m, 1H), 2.29 (s, 3H), 1.18 (d, 6H, J=6.9 Hz); .sup.13C NMR (75
MHz, DMSO-d.sub.6) .delta. 165.1, 158.2, 150.2, 149.6, 142.2,
139.8, 136.8, 136.7, 135.0, 132.4, 130.0, 125.0, 124.6, 122.5,
120.7, 118.8, 118.2, 114.2, 66.6, 28.7, 23.5, 19.5; MS (ESI) m/z
393 [M-H].sup.-.
[0344] (c) A solution of
3-(6-chloropyridin-3-yl)methoxy-N-(4-isopropyl-3--
methyl-phenyl)-benzamide, L-1b, (300 mg, 0.76 mmol, 1.0 eq),
4-methoxybenzylamine (Aldrich, 0.12 mL, 0.91 mmol, 1.2 eq),
palladium acetate (Strem, 6.8 mg, 0.03 mmol, 4 mol %), and CyMAP1
(J. Am. Chem. Soc. 1998, 120, 9722-23, 11.8 mg, 0.03 mmol, 4 mol %)
in dioxane (10 mL) was treated with sodium-tert-butoxide (169 mg,
1.82 mmol, 1.4 eq). The resultant orange solution was warmed to
80.degree. C. for 18 hours. The reaction mixture was poured into
50% brine (100 mL) and extracted with ethyl acetate (3.times.75
mL). The combined organic extracts were washed with brine (200 mL),
dried over magnesium sulfate, filtered through a silica gel plug,
and concentrated under reduced pressure to give a yellow oil (330
mg). The crude product was purified by radial chromatography over
silica gel using 45% ethyl acetate/cyclohexane to give, from MTBE,
N-(4-isopropyl-3-methyl-phenyl)-3-[6-{(4-methoxybenzyl)amino}pyridin-3-yl-
methoxy]-benzamide, L-1c, as a white solid (189 mg, 50%): HPLC
R.sub.t=14.7 min.; TLC R.sub.f=0.4 (4% methanol/dichloromethane);
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 9.43 (s, 1H), 7.74 (s,
1H), 7.27-7.18 (m, 3H), 7.02-6.82 (m, 6H), 6.70-6.62 (m, 4H), 6.41
(d, 1H, J=8.5 Hz), 4.79 (s, 2H), 4.32 (d, 2H, J=5.5 Hz), 3.69 (s,
3H), 2.97-2.90 (m, 1H), 2.13 (s, 3H), 1.05 (d, 6H, J=6.7 Hz);
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. 169.9, 158.4, 158.3,
157.0, 147.4, 144.8, 140.4, 138.0, 137.2, 135.7, 132.7, 129.0,
128.8, 125.8, 125.3, 120.8, 119.2, 116.7, 115.5, 113.9, 108.2,
55.4, 50.7, 44.0, 28.7, 23.3, 19.2; MS (ESI) m/z 494
[M-H].sup.-.
[0345] (d) A solution of
N-(4-isopropyl-3-methyl-phenyl)-3-[6-{(4-methoxyb-
enzyl)amino}pyridin-3-ylmethoxy]-benzamide, L-1c, (120 mg, 0.24
mmol) in trifluoroacetic acid (6 mL) was stirred at room
temperature. After 18 hours, the resultant cherry red solution was
concentrated under reduced pressure and extracted with 50% sodium
bicarbonate (25 mL) and ethyl acetate (3.times.25 mL). The combined
organic extracts were washed with 5% sodium bicarbonate (50 mL),
brine (50 mL), dried over magnesium sulfate, and concentrated under
reduced pressure to give a clear oil (132 mg). The crude product
was purified by radial chromatography over silica gel using 5%
methanol/chloroform with 0.1% ammonium hydroxide to give,
3-(6-amino-pyridin-3-yl)methoxy-N-(4-isopropyl-3-methyl-phenyl)-benzamide-
, L-1, as a white solid (75 mg, 82%): mp 78-81.degree. C.; HPLC
R.sub.t=12.1 min.; TLC R.sub.f=0.4 (8% methanol/chloroform);
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 9.32 (s, 1H), 7.54 (d,
1H, J=2.0 Hz), 7.13 (dd, 1H, J=8.4, 2.4 Hz), 6.90-6.71 (m, 2H),
6.58 (d, 1H, J=1.5 Hz), 6.54-6.47 (m, 4H), 6.23 (d, 1H, J=8.5 Hz),
5.70 (s, 2H), 4.66 (s, 2H), 2.94-2.78 (m, 1H), 2.02 (s, 3H), 0.94
(d, 6H, J=6.8 Hz); MS (ESI) m/z 494 [M-H].sup.-. Anal. calcd for
C.sub.23H.sub.25N.sub.3O.sub.2.0.4 hexane: C, 74.42; H, 7.52; N,
10.25. Found: C, 74.09; H, 7.49; N, 10.00.
EXAMPLE M-1
3-(6-Aminopyridin-3-yl)methoxy-N-(2-methyl-quinolin-6-yl)-benzamide
[0346] 180
[0347] (a) A mixture of 6-chloronicotinic acid (Aldrich, 5.0 g,
31.6 mmol, 1.0 eq), water (10 mL) and ethanol (35 mL) was treated
with sodium azide (2.15 g, 33.8 mmol, 1.1 eq). The resulting orange
solution was heated to 75.degree. C. After 18 h, the solvent was
removed under reduced pressure and the resultant white slurry was
repeatedly evaporated with toluene to give the crude
6-azido-nicotinic acid/NaCl mixture, M-1a, as a white solid (6.9 g,
98%): TLC R.sub.f 0.2 (6% methanol/dichloromethane w/ 0.1% Acetic
acid); .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 9.41 (s, 1H),
7.98 (s, 1H), 7.97 (s, 1H).
[0348] (b) A mixture of 6-azidonicotinic acid/NaCl, M-1a, (6.9 g,
31.1 mmol), and thionyl chloride (100 mL) was heated to 75.degree.
C. After 2 hours, the crude acid chloride hydrochloride salt was
obtained by removing the thionyl chloride under pressure. The
resultant slurry was evaporated from toluene and then added to a
slurry of isopropanol (100 mL) and sodium borohydride at
-10.degree. C. The resultant yellow slurry was warmed to room
temperature over several hours and stirred at room temperature for
12 h. The reaction mixture was poured into water (500 mL) and
extracted with ethyl acetate (3.times.500 mL). The combined organic
extracts were washed with brine (200 mL), dried over magnesium
sulfate, filtered and concentrated under reduced pressure to give a
yellow solid (2.3 g). The crude product was purified by radial
chromatography over silica gel using 1-3% methanol/dichloromethane
to give (6-azidopyridin-3-yl)-methanol, M-1b, as a white solid (850
mg, 18%): TLC R.sub.f=0.4 (5% methanol/chloroform); .sup.1H NMR
(500 MHz, DMSO-d.sub.6) .delta. 9.14 (s, 1H), 8.19 (d, 1H, J=9.0
Hz), 7.84 (dd, 1H, J=9.4, 1.0 Hz), 5.67 (t, 1H, J=5.6 Hz), 4.66 (d,
2H, J=5.6 Hz).
[0349] (c) A solution of (6-azidopyridin-3-yl)-methanol, M-1b, (350
mg, 2.3 mmol, 1.0 eq) in THF (25 mL) was cooled to -78.degree. C.
and treated with mesyl chloride (265 .mu.L, 3.4 mmol, 1.5 eq).
After 60 minutes at -78.degree. C., the clear solution was treated
with triethylamine (0.70 mL, 5.1 mmol, 2.2 eq), and the reaction
mixture was warmed to -20.degree. C. over 2.5 hours. The resultant
cloudy reaction mixture was monitored by TLC (3%
methanol/dicbloromethane), which gave only product (R.sub.f 0.4)
and no starting material (R.sub.f 0.2). To obtain an analytical
sample, an aliquot of the reaction mixture was diluted in 25%
ammonium acetate and extracted with ethyl acetate. The organic
extract was washed with water, brine, dried over magnesium sulfate,
filtered and concentrated under reduced pressure to give
(6-azidopyridin-3-yl)methyl methanesulfonate, M-1c, as a tan solid:
TLC R.sub.f=0.6 (5% methanol/chloroform); .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. 9.5 (s, 1H), 8.30 (d, 1H, J=9.4 Hz), 7.94 (d,
1H, J=9.6 Hz), 5.46 (s, 2H), 3.35 (s, 3H).
[0350] (d) A solution of 3-acetoxybenzoic acid (Aldrich, 1.8 g, 9.9
mmol, 1.0 eq) and 6-amino-2-methylquinoline (Avocado, 1.6 g, 9.9
mmol, 1.0 eq) in ethyl acetate (50 mL) was treated with a solution
of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.3
g, 11.8 mmol, 1.2 eq) in dichloromethane (50 mL). After 18 h, the
resultant tan slurry was poured into 5% sodium bicarbonate (200 mL)
and extracted with 10% isopropyl alcohol/chloroform (3.times.150
mL). The combined organic extracts were washed with brine (200 mL),
dried over magnesium sulfate, filtered through a silica gel plug
and concentrated under reduced pressure to give a yellow solid (2.8
g). The crude product was purified by radial chromatography over
silica gel using 1-2% methanol/dichloromethane to give an unpure
product as a yellow solid (2.6 g). The unpure product was washed
with MTBE and the solids were collected to give
3-acetoxy-N-(2-methyl-quinolin-6-yl)benzamide, M-1d, as a light
yellow solid (2.3 g, 84%): HPLC R.sub.t=11.2 min.; TLC R.sub.f=0.3
(3% methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.40 (s, 1H), 8.29 (d, 1H, J=2.2 Hz), 8.04 (d, 1H, J=8.5
Hz), 7.83-7.72 (m, 3H), 7.59 (t, 1H, J=1.8 Hz), 7.45 (t, 1H, J=7.9
Hz), 7.24 (d, 1H, J=8.4 Hz), 2.48 (s, 3H), 2.16 (s, 3H); MS (ESI)
m/z 321 [M+H].sup.+.
[0351] (e) A solution of
3-acetoxy-N-(2-methyl-quinolin-6-yl)benzamide, M-1d, (2.2 g, 6.9
mmol, 1.0 eq) in methanol (65 mL) and THF (60 mL) was treated with
a solution of potassium carbonate (4.8 g, 34.5 mmol, 5.0 eq) in
water (45 mL). After 3.0 h, the cloudy reaction mixture was
concentrated under reduced pressure and extracted with water (100
mL) and ethyl acetate (3.times.100 mL). The combined organic
extracts were washed with brine (200 mL), dried over magnesium
sulfate and concentrated under reduced pressure to give a light
yellow solid (2.0 g). The crude product was washed with
dichloromethane and the solids were collected to give
3-hydroxy-N-(2-methyl-quinolin-6-yl)-benzamide, M-1e, as a light
yellow solid (1.6 g, 83%): HPLC R.sub.t=9.5 nm.; TLC R.sub.f=0.3
(5% methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.43 (s, 1H), 9.78 (s, 1H), 8.47 (d, 1H, J=2.2 Hz), 8.20
(d, 1H, J=8.4 Hz), 8.00-7.87 (m, 2H), 7.44-7.32 (m, 4H), 7.01-6.98
(m, 1H), 2.64 (s, 3H); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
166.2, 157.7, 157.6, 144.8, 136.8, 136.6, 136.1, 129.8, 128.8,
126.7, 124.4, 122.8, 119.0, 118.6, 116.6, 114.9, 25.0; MS (ESI) m/z
279 [M+H].sup.+.
[0352] (f) A mixture of
3-hydroxy-N-(2-methyl-quinolin-6-yl)-benzamide, M-1e, (0.64 g, 2.3
mmol, 1.0 eq) and cesium carbonate (3.0 g, 9.2 mmol, 4.0 eq) in
acetone (45 mL) was treated with unpurified
(6-azidopyridin-3-yl)methyl methanesulfonate, M-1c, (524 mg, 2.3
mmol, 1.0 eq). After 18 h at 50.degree. C., the resultant pink
slurry was poured into 5% sodium bicarbonate (400 mL) and extracted
with 5% isopropyl alcohol/chloroform (3.times.300 mL). The combined
organic extracts were washed with brine (300 mL), dried over
magnesium sulfate and concentrated under reduced pressure to give a
light yellow solid (0.99 g). The crude product was washed with
diethyl ether and the solids were collected to give
3-(6-azidopyridin-3-yl)methoxy-N-(2-methyl-quinoli-
n-6-yl)-benzamide, M-1f, as a light yellow solid (0.87 g, 92%):
HPLC R.sub.t=11.7 min.; TLC R.sub.f=0.5 (5%
methanol/dichloromethane); .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. 10.51 (s, 1H), 9.53 (s, 1H), 8.46 (d, 1H, J=2.0 Hz), 8.30
(d, 1H, J=9.2 Hz), 8.21 (d, 1H, J=9.0 Hz), 8.02-7.98 (m, 2H), 7.91
(d, 1H, J=9.1 Hz), 7.72 (s, 1H), 7.66 (d, 1H, J=7.8 Hz), 7.54 (t,
1H, J=8.0 Hz), 7.40 (d, 1H, J=8.4 Hz), 7.36 (dd, 1H, J=8.2, 2.0
Hz), 5.40 (s, 2H), 2.51 (s, 3H); MS (ESI) m/z 411 [M+H].sup.+.
[0353] (g) A solution of
3-(6-azidopyridin-3-yl)methoxy-N-(2-methyl-quinol-
in-6-yl)-benzamide, M-1f, (58 mg, 0.14 mmol, 1.0 eq) in anhydrous
ethanol (20 mL) was treated with tin(II) chloride dihydrate (158
mg, 0.70 mmol, 5.0 eq) and warmed to 70.degree. C. After 18 h, the
light yellow solution was concentrated under reduced pressure and
treated with a saturated sodium bicarbonate solution (100 mL). The
aqueous layer was extracted with ethyl acetate (3.times.100 mL) and
the combined organic extracts were washed with brine (200 mL),
dried over magnesium sulfate, filtered and concentrated under
reduced pressure to give a white foam (60 mg). The crude product
was purified by radial chromatography over silica gel using 5%
methanol/chloroform to give
3-(6-aminopyridin-3-yl)methoxy-N-(2-methyl-
-quinolin-6-yl)-benzamide, M-1, as a white solid (23 mg, 42%): mp
189-192.degree. C.; HPLC R.sub.t=11.4 min.; TLC R.sub.f=0.3 (5%
methanol/chloroform); .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
10.48 (s, 1H), 8.49 (d, 1H, J=2.0 Hz), 8.22 (d, 1H, J=8.5 Hz), 8.05
(d, 1H, J=1.8 Hz), 8.00 (dd, 1H, J=9.1, 2.2 Hz), 7.92 (d, 1H, J=9.1
Hz), 7.64-7.41 (m, 4H), 7.41 (d, 1H, J=8.4 Hz), 7.26 (dd, 1H,
J=8.2, 1.9 Hz), 6.49 (d, 1H, J=8.4 Hz), 6.05 (s, 2H), 5.01 (s, 2H),
2.66 (s, 3H); MS (ESI) m/z 385 [M+H].sup.+.
[0354] Anal. calcd for C.sub.23H.sub.20N.sub.4O.sub.2.0.5H.sub.2O:
C, 70.21; H, 5.38; N, 14.24. Found: C, 70.42; H, 5.34; N,
13.85.
EXAMPLE N-1
3-(6-Acetylaminopyridin-3-yl)methoxy-N-(2-methyl-quinolin-6-yl)-benzamide
[0355] 181
[0356] A cloudy suspension of
3-(6-amino-pyridin-3-yl)methoxy-N-(2-methyl--
quinolin-6-yl)-benzamide, M-1, (40 mg, 0.10 mmol) in acetic
anhydride (5 mL), THF (5 mL) and dichloromethane (3 mL) was stirred
at room temperature. After 18 h, the resultant yellow solution was
concentrated under reduced pressure and chased with toluene to give
a yellow solid (56 mg). The crude product was purified by radial
chromatography over silica gel using 3-10% methanol/dichloromethane
to give 3-[6-(acetylaminopyridin-
-3-yl)methoxy-N-(2-methyl-quinolin-6-yl)-benzamide, N-1, as a white
solid (40 mg, 91%): HPLC R.sub.t=11.5 min.; TLC R.sub.f=0.5 (8%
methanol/chloroform); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
10.63 (s, 1H), 10.56 (s, 1H), 8.54 (s, 1H), 8.50 (s, 1H), 8.29 (d,
1H, J=8.4 Hz), 8.18 (d, 1H, J=8.6 Hz), 8.07-7.95 (m, 3H), 7.71-7.67
(m, 2H), 7.56 (t, 1H, J=8.0 Hz), 7.47 (d, 1H, J=8.1 Hz), 7.35 (d,
1H, J=7.2 Hz), 5.25 (s, 2H), 2.71 (s, 3H), 2.71 (s, 3H); MS (ESI)
m/z 427 [M+H].sup.+. Anal. calc'd for
C.sub.25H.sub.22N.sub.4O.sub.3.0.1H.sub.2O: C, 70.11; H, 5.23; N,
13.08. Found: C, 69.71; H, 5.33; N, 12.82.
EXAMPLE N-2
3-(6-Acetylaminopyridin-3-yl)methoxy-N-(4-Isopropyl-3-methyl-phenyl)-benza-
mide
[0357] 182
[0358] A clear solution of
3-(6-amino-pyridin-3-yl)methoxy-N-(4-isopropyl--
3-methyl-phenyl)-benzamide, L-1, (30 mg, 0.08 mmol) in acetic
anhydride (1.0 mL) was stirred at room temperature for 18 h. The
resultant clear solution was concentrated under reduced pressure
and chased with toluene to give a clear oil. The crude product was
extracted with 5% sodium bicarbonate (25 mL) and ethyl acetate
(3.times.25 mL). The combined ethyl acetate extracts were dried
using brine (25 mL) and magnesium sulfate to give a clear oil (30
mg), which was identified as 3-(6-diacetylamino-pyri-
din-3-yl)methoxy-N-(4-isopropyl-3-methyl-phenyl)-benzamide by MS
(ESI) (m/z 460). After purification by radial chromatography over
silica gel, the diacetylated compound (24 mg, 0.05 mmol, 1.0 eq)
was dissolved in methanol (0.5 mL) and treated with calcium
carbonate (10 mg, 0.1 mmol, 2.1 eq) and water (0.5 mL). The
resultant white slurry was heated at 60.degree. C. for 18 hours.
The reaction mixture was poured into water (25 mL) and extracted
with ethyl acetate (3.times.25 mL). The combined organic extracts
were washed with brine (50 mL), dried over magnesium sulfate, and
concentrated under reduced pressure to give a clear oil (40 mg).
The crude product was purified by radial chromatography over silica
gel using 2-5% methanol/dichloromethane to give
3-(6-acetylaminopyridin-3-
-yl)methoxy-N-(4-isopropyl-3-methyl-phenyl)-benzamide, N-2, as a
white solid (20 mg, 61%): mp 98-101.degree. C.; HPLC R.sub.t=12.4
min.; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.51 (s, 1H),
9.54 (s, 1H), 8.19 (s, 1H), 8.06 (d, 1H, J=8.2 Hz), 7.72 (d, 1H,
J=8.3 Hz), 7.11-6.97 (m, 3H), 6.82-6.71 (m, 4H), 5.04 (s, 2H),
3.08-3.01 (m, 1H), 2.22 (s, 3H), 2.13 (s, 3H), 1.13 (d, 6H, J=6.7
Hz); MS (ESI) m/z 418 [M+H].sup.+. Anal. calc'd for
C.sub.25H.sub.27N.sub.3O.sub.3.0.4 hexane .cndot.0.5H.sub.2O: C,
71.39; H, 7.35; N, 9.12. Found: C, 71.11; H, 7.48; N, 8.70.
EXAMPLE O-1
4-Fluoro-N-(1,2,3,4-tetrahydroquinolin-6-yl)-3-(isoquinolin-4-yl-methoxy)--
benzamide Bistrifluoroacetic Acid Salt
[0359] 183
[0360] (a) To a stirred mixture of 4-fluoro-3-hydroxy-benzoic acid
(0.73 g, 4.7 mmol) and Cs.sub.2CO.sub.3 (4.58 g, 14.1 mmol) in
dioxane/H.sub.2O (1:1, 20 ml) was added 1.0 g (4.7 mmol) of
4-chloromethyl-isoquinoline.cn- dot.HCl, K-1c. After heating at
65.degree. C. for 24 hrs, the solvent was removed, water was added,
the pH was adjusted to 6, and the solution was extracted with ethyl
acetate (30 ml.times.3). The combined extracts were washed with 1N
HCl. A precipitate formed and was filtered and dried to provide
0.41 g (29%) of 4-fluoro-3-(isoquinolin-4-yl)methoxy-benzoic acid
hydrochloride, O-1a, as a solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 9.84 (s, 1H), 8.79 (s, 1H), 8.54 (d, J=8.1
Hz, 1H), 8.44 (d, J=8.36 Hz, 1H), 8.21 (m, 1H), 8.02 (m, 2H), 7.65
(m, 1H), 7.40 (dd, J=11.00, 8.46 Hz, 1H), 5.865 (s, 2H).
[0361] (b) To an ice cooled solution of 0.45 g (1.85 mmol) of
6-(2,2,2-trifluoroacetylamino)-1,2,3,4-tetrahydroquinoline (Forbes,
et al., J. Med. Chem., 38, 2524 (1995)) in THF (20 ml) was added
di-t-butyl-dicarbonate (0.89 g, 4.08 mmol). After refluxing for 24
h, the solvent was removed and the residue was purified on silica
gel using a gradient of 0% to 2% ethyl acetate in dichloromethane
as eluant to obtain 0.42 g (66%) of
1-(tert-butoxycarbonyl)-6-(2,2,2-trifluoroacetylamino)-1,-
2,3,4-tetrahydroquinoline, O-1b, as a solid: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.78 (br s, 1H), 7.71 (d, J=8.9 Hz, 1H), 7.43
(d, J=2.6 Hz, 1H), 7.19 (dd, J=8.9, 2.6 Hz, 1H), 3.70 (m, 2H), 2.77
(t, J=6.5 Hz, 2H), 1.90 (t, J=6.4 Hz, 2H), 1.52 (s, 9H).
[0362] (c) To stirred solution of 0.41 g (1.19 mmol) of
1-(tert-butoxycarbonyl)-6-(2,2,2-trifluoro-acetylamino)-1,2,3,4-tetrahydr-
oquinoline, O-1b, in methanol (20 ml) was added K.sub.2CO.sub.3
(0.25 g, 1.79 mmol). After refluxing for 24 h, the methanol was
removed, followed by addition of water and ethyl acetate. The
layers were separated, and the aqueous layer was extracted with
ethyl acetate. The combined extracts were washed with brine, dried
over MgSO.sub.4, and concentrated to leave 0.228 g (77%) of
6-amino-1-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroquinol- ine,
O-1c, as an oil: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.38 (d,
J=8.5 Hz, 1H), 6.47 (dd, J=8.7, 2.7 Hz, 1H), 6.39 (m, 1H), 3.64 (m,
2H), 3.40 (brs, 1H), 2.65 (t, J=6.6 Hz, 2H), 1.86 (t, J=6.2 Hz,
2H), 1.50, (s, 9H).
[0363] (d) A solution of 0.35 g (1.17 mmol) of
4-fluoro-3-(isoquinolin-4-y- l)methoxy-benzoic acid hydrochloride,
O-1a, in SOCl.sub.2 (5 ml) was stirred and heated at 60.degree. C.
for 3 h. The thionyl chloride was removed under reduced pressure to
provide crude acid chloride O-1d, which was dissolved in
dichloromethane (15 ml) under argon. The solution was cooled to
0.degree. C. followed by addition of 0.260 g (1.07 mmol) of
6-amino-1-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroquinoline, O-1c,
and diisopropylethylamine (0.3 g, 2.34 mmol). After stirring for 24
hrs, dichloromethane (15 ml) was added, and the solution was washed
with sat. NaHCO.sub.3, and dried over sodium sulfate and
concentrated. The residue was purified on silica gel using a
gradient of 0% to 5% ethyl acetate in dichloromethane as eluant to
obtain 0.190 g (30%) of
4-fluoro-N-{1-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroquinolin-6-yl}-3-(i-
soquinolin-4-yl-methoxy)-benzamide, O-1e.
[0364] (e) A solution of O-1e (0.185 g, 0.35 mmol) in 4N HCl in
dioxane (5 ml) was stirred for 6 hrs at 0.degree. C. The solvent
was removed under reduced pressure to give 0.123 g (82%) of
4-Fluoro-N-(1,2,3,4-tetrahydroq-
uinolin-6-yl)-3-(isoquinolin-4-yl-methoxy)-benzamide hydrochloride
as a solid. This product was further purified on semi-preparative
C18-reverse phase HPLC eluting with 5 to 95% acetonitrile/water
containing 0.1% trifluoroacetic acid to provide
4-fluoro-N-(1,2,3,4-tetrahydroquinolin-6--
yl)-3-(isoquinolin-4-yl-methoxy)-benzamide bistrifluoroacetic acid
salt, O-1: .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.59 (s,
1H), 9.81 (s, 1H), 8.83 (s, 1H), 8.52 (d, 1H, J=8.37 Hz) 8.45 (d,
1H, J=8.59 Hz), 8.24-8.17 (m, 2H), 8.01 (dd, 1H, J=7.35 Hz, J=7.6
Hz), 7.79-7.70 (m, 3H), 7.43 (dd, 1H, J=10.87 Hz, J=10.7 Hz), 7.25
(d, 1H, J=8.46 Hz), 5.93 (s, 2H), 3.73-3.64 (m, 1H), 3.52-3.45 (m,
1H), 3.37-3.35 (m, 1H), 2.87-2.83 (m, 2H), 2.02-1.94 (m, 2H). MS
(ESI) m/z 428 [M].sup.+. Anal. calc'd for
C.sub.26H.sub.22FN.sub.3O.sub.2.2 CF.sub.3CO.sub.2H.0.8H.sub.2O: C,
53.78; H, 3.85; N, 6.27. Found: C, 53.58; H, 4.00; N, 6.20.
EXAMPLE O-2
N-(2,2-Difluoro-benzo[1,3]dioxol-4-yl)-3-(2-isoquinolin-4-yl-ethyl)-benzam-
ide Trifluoro-Acetic Acid Salt
[0365] 184
[0366] Example O-2 was prepared in a similar manner to that
described for O-1, except that
4-amino(2,2-difluoro-benzo[1,3]dioxole was used in place of
6-amino-1-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroquinoline, O-1c,
and 3-{2-(isoquinolin-4-yl)ethyl}benzoic acid, S-1e (from example
S-1 below), was used in place of
4-fluoro-3-(isoquinolin-4-yl)methoxy-benzoic acid, O-1a, in step
(d), and the final deprotection step was not needed: .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 10.70 (s, 1H), 9.88 (s, 1H),
8.63-8.51 (m, 3H), 8.26 (dd, 1H, J=8.3 Hz, J=8.5 Hz), 8.05 (m, 2H),
7.87 (m, 1H), 7.62-7.43 (m, 2H), 7.31-7.25 (m, 3H), 3.62-3.51 (m,
2H), 3.17-3.09 (m, 2H). MS (ESI) m/z 433 [M].sup.+. Anal. calc'd
for C.sub.25H.sub.18 F.sub.2N.sub.2O.sub.3.C.sub.2F.sub.3OOH: C,
59.35; H, 3.50; N, 5.31. Found: C, 59.35; H, 3.60; N, 5.12.
EXAMPLE O-3
4-Fluoro-N-(2-methyl-1,2,3,4-tetrahydroquinolin-6-yl)-3-(isoquinolin-4-yl--
methoxy)-benzamide Bistrifluoroacetic Acid Salt
[0367] 185
[0368] Example O-3 was prepared in a similar manner to that
described for O-1, except that
6-amino-1-(tert-butoxycarbonyl)-2-methyl-1,2,3,4-tetrahy-
droquinoline was used in place of
6-amino-1-(tert-butoxycarbonyl)-1,2,3,4-- tetrahydroquinoline in
step (d): .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.15 (s,
1H), 9.54 (s, 1H), 8.73 (s, 1H), 8.35-8.26 (m, 2H), 8.04-7.98 (m,
2H), 7.86 (dd, 1H, J=7.12 Hz, J=7.08 Hz), 7.67-7.62 (m, 1H),
7.52-7.49 (m, 2H), 7.41 (dd, 1H, J=10.96 Hz, J=10.99 Hz), 6.90 (d,
1H, J=8.37 Hz), 5.78 (s, 2H), 3.46 (m, 1H), 2.87-2.68 (m, 2H),
2.01-1.97 (m, 1H), 1.69-1.63 (m, 1H), 1.27 (d, 3H, J=6.41 Hz). MS
(ESI) m/z 442 [M].sup.+. Anal. calc'd for
C.sub.27H.sub.24FN.sub.3O.sub.2.2 CF.sub.3CO.sub.2H: C, 55.61; H,
3.91; N, 6.28. Found: C, 55.51; H, 3.88; N, 6.22.
EXAMPLE P-1
N'-{4-[3-(4-Isopropyl-3-methyl-phenylcarbamoyl)-phenoxy
[0369] 186
[0370] (a) A solution of 4-bromoisoquinoline (Aldrich, 7.7 g, 37.0
mmol, 1.0 eq) and perrhenic acid (70% in H.sub.2O, 33 .mu.L, 0.2
mmol, 0.5 mol %) in dichloromethane (20 mL) was treated with
bis(trimethylsilyl)peroxid- e (Gelest, 9.9 g, 55.5 mmol, 1.5 eq).
After 18 hours, the resultant yellow suspension was cooled to
0.degree. C. and diluted with cyclohexane (30 mL). The solids were
collected and washed with cold cyclohexane to give
4-bromoisoquinoline N-oxide, P-1a, as a yellow solid (7.1 g, 86%):
HPLC R.sub.t=7.7 min.; TLC R.sub.f=0.5(5%
methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 8.97 (s, 1H), 8.58 (d, 1H, J=1.7 Hz), 7.97-7.87 (m, 2H),
7.74-7.67 (m, 2H); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
138.6, 135.5, 130.6, 130.2, 129.6, 127.4, 126.0, 125.7, 120.1; MS
(ESI) m/z 224/226 [M+H].sup.+.
[0371] (b) A yellow suspension of 4-bromoisoquinoline N-oxide,
P-1a, (6.9 g, 30.8 mmol. 1.0 eq) in 1,2-dichloroethane (60 mL) was
treated with phosphorus oxychloride (Aldrich, 9.0 mL, 96.4 mmol,
1.8 eq) and warmed to 80.degree. C. After 1.5 hours, the resultant
green suspension was carefully poured into a cold solution of 50%
saturated sodium bicarbonate (500 mL) and the aqueous layer was
extracted with diethyl ether (3.times.300 mL). The combined organic
extracts were washed with water (200 mL), brine (200 mL), dried
over magnesium sulfate, and concentrated under reduced pressure to
give a tan solid (6.8 g). The crude product was dissolved in a
minimal amount of dichloromethane and purified by flash
chromatography over silica gel using 5% ether/cyclohexane to give
4-bromo-1-chloro-isoquinoline, P-1b, as a white solid (5.7 g, 77%):
HPLC R.sub.t=15.4 min.; TLC R.sub.f=0.4 (5% ether/cyclohexane);
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 8.68 (s, 1H), 8.42 (d,
1H, J=8.4 Hz), 8.26 (d, 1H, J=8.3 Hz), 8.15 (t, 1H, J=7.6 Hz), 8.02
(t, 1H, J=7.6 Hz); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
150.4, 143.0, 135.8, 133.8, 130.8, 127.3, 126.8, 126.5, 119.0; MS
(ESI) m/z 242/244 [M+H].sup.+.
[0372] (c) 1-Chloro-isoquinoline-4-carbaldehyde, P-1c, which was
obtained as a white solid in a 95% yield, was prepared in a similar
manner to that described for isoquinoline-4-carbaldehyde, K-1a, in
example K-1, except that 4-bromo-1-chloro-isoquinoline, P-1b, was
used in place of 4-bromoisoquinoline: HPLC R.sub.t=11.9 min.; TLC
R.sub.f=0.6 (20% ethyl acetate/cyclohexane); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.40 (s, 1H), 9.15 (d, 1H, J=8.5 Hz), 8.93
(s, 1H), 8.46 (d, 1H, J=8.5 Hz), 8.12 (t, 1H, J=7.1 Hz), 8.00 (t,
1H, J=7.8 Hz); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. 193.6,
156.5, 151.4, 134.8, 133.9, 130.4, 126.9, 126.0, 124.9, 124.6; MS
(ESI) m/z 192 [M+H].sup.+.
[0373] (d) (1-Chloro-isoquinolin-4-yl)-methanol, P-1d, which was
obtained as a white solid in a 96% yield, was prepared in a similar
manner to that described for isoquinolin-4-yl-methanol, K-1b,
except that 1-chloro-isoquinoline-4-carbaldehyde, P-1c, was used in
place isoquinoline-4-carbaldehyde, K-1a: HPLC R.sub.t=9.0 min.; TLC
R.sub.f=0.2 (2% methanol/dichloromethane); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 8.52 (d, 1H, J=8.5 Hz), 8.50 (s, 1H), 8.42
(d, 1H, J=8.3 Hz), 8.15 (t, 1H, J=7.6 Hz), 8.05 (t, 1H, J=7.6 Hz),
5.71 (br s, 1H), 5.15 (s, 2H); .sup.13C NMR (75 MHz, DMSO-d.sub.6)
.delta. 150.1, 140.0, 136.0, 131.9, 131.8, 129.3, 126.3, 125.8,
124.6, 59.0; MS (ESI) m/z 194 [M+H].sup.+.
[0374] (e)
3-(1-Chloro-isoquinolin-4-ylmethoxy)-N-(4-isopropyl-3-methyl-ph-
enyl)-benzamide, P-1e, which was obtained as a white solid in a 64%
yield, was prepared in a similar manner to that described for
3-(6-chloropyridin-3-yl)methoxy-N-(4-isopropyl-3-methyl-phenyl)-benzamide-
, L-1b, except that (1-chloro-isoquinolin-4-yl)-methanol, P-1d, was
used in place of (6-chloropyridin-3-yl)-methanol, L-1a: HPLC
R.sub.t=18.0 min.; TLC R.sub.f=0.5 (5% ethyl acetate/45%
dichloromethane/cyclohexane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.06 (s, 1H), 8.52 (s, 1H), 8.38 (d, 1H, J=8.5 Hz), 8.25
(d, 1H, J=8.3 Hz), 8.01 (t, 1H, J=7.6 Hz), 7.90 (t, 1H, J=8.2 Hz),
7.68 (s, 1H), 7.61-7.46 (m, 4H), 7.35 (d, 1H, J=7.9 Hz), 7.20 (d,
1H, J=8.4 Hz), 5.65 (s, 2H), 3.12-3.05 (m, 1H), 2.29 (s, 3H), 1.17
(d, 6H, J=6.9 Hz); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
165.1, 158.4, 151.5, 142.2, 141.9, 136.8, 136.7, 136.3, 135.0,
132.6, 130.0, 129.8, 127.1, 126.6, 126.1, 125.0, 124.7, 122.5,
120.8, 118.9, 118.3, 114.3, 65.6, 28.7, 27.2 MS (ESI) m/z 443
[M-H].sup.-.
[0375] (f) A mixture of
3-(1-chloro-isoquinolin-4-ylmethoxy)-N-(4-isopropy-
l-3-methyl-phenyl)-benzamide, P-1e, (50 mg, 0.11 mmol, 1.0 eq),
tert-butyl carbazate (75 mg, 0.56 mmol, 5.0 eq) and
para-toluenesulfonic acid monohydrate (31 mg, 0.16 mmol, 1.5 eq) in
isopropyl alcohol (1.5 mL) was warmed to 60.degree. C. After 4.5 h
the resultant white suspension was poured into 5% sodium
bicarbonate (25 mL) and extracted with ethyl acetate (3.times.25
mL). The combined organic extracts were washed sequentially with
water (25 mL), sodium citrate (0.5 M, pH 4.5, 25 mL), water (25 mL)
and brine (25 mL). The organic extracts were dried over magnesium
sulfate, filtered, and concentrated under reduced pressure to give
a clear oil (90 mg). The crude product was purified by radial
chromatography over silica gel using 2-3% methanol/dichloromethane
to give, from methyl tert-butyl ether,
N'-{4-[3-(4-isopropyl-3-methyl-phenyl-
carbamoyl)-phenoxymethyl]-isoquinolin-1-yl}-hydrazinecarboxylic
acid tert-butyl ester, P-1, as a white solid (55 mg, 93%): mp
125-129.degree. C.; HPLC R.sub.t=16.5 min.; TLC R.sub.f=0.5 (4%
methanol/dichloromethane)- ; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.04 (s, 1H), 9.35 (s, 1H), 8.78 (s, 1H), 8.33 (d, 1H,
J=8.3 Hz), 8.10 (s, 1H), 7.96 (d, 1H, J=8.3 Hz), 7.76 (t, 1H, J=7.6
Hz), 7.64-7.27 (m, 7H), 7.20 (d, 1H, J=8.4 Hz), 5.41 (s, 2H),
3.09-3.05 (m, 1H), 2.29 (s, 3H), 1.40 (s, 9H), 1.17 (d, 6H, J=6.8
Hz); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. 165.8, 159.4,
157.2, 155.6, 143.8, 141.5, 137.1, 136.3, 136.1, 135.6, 131.2,
130.2, 127.1, 125.7, 124.2, 122.4, 122.3, 119.7, 119.0, 118.6,
118.5, 117.8, 113.9, 81.9, 67.3, 29.3, 28.6, 27.3, 23.6, 19.8; MS
(ESI) m/z 541 [M+H].sup.+. Anal. calc'd for
C.sub.32H.sub.36N.sub.4O.sub.4.0.25 hexane: C, 71.57; H, 7.08; N,
9.97. Found: C, 71.38; H, 7.18; N, 9.66.
EXAMPLE Q-1
N-(4-Isopropyl-3-methyl-phenyl)-3-{1-[N'-(3-methoxy-benzylidene)-hydrazino-
]-isoquinolin-4-ylmethoxy}-benzamide
[0376] 187
[0377] A solution of
N'-{4-[3-(4-isopropyl-3-methyl-phenylcarbamoyl)-pheno-
xymethyl]-isoquinolin-1-yl}-hydrazinecarboxylic acid tert-butyl
ester, P-1, (94 mg, 0.21 mmol, 1.0 eq) in dichloromethane (2.0 mL)
was treated with trifluoroacetic acid (0.5 mL). After 1.5 hours,
the resultant yellow solution was carefully poured into 5% sodium
bicarbonate (50 mL) and extracted with ethyl acetate (3.times.25
mL). The combined organic extracts were washed with brine (25 mL),
dried over magnesium sulfate, and concentrated under reduced
pressure to give a yellow solid (97 mg). The crude product was
dissolved in ethyl alcohol (4 mL) and treated with acetic acid (3
drops), and 3-methoxybenzaldehyde (Aldrich, 40 .mu.L, 0.32 mmol,
1.5 eq). After 18 hours, the resultant yellow suspension was poured
into 5% sodium bicarbonate (25 mL). The aqueous layer was diluted
with brine (15 mL) and extracted with ethyl acetate (3.times.25
mL). The combined organic extracts were washed with brine (50 mL),
dried over magnesium sulfate, and concentrated under reduced
pressure to give a yellow residue (128 mg). The crude product was
purified by radial chromatography over silica gel using 1-2%
methanol/dichloromethane to give
N-(4-isopropyl-3-methylphenyl)-3-{1-[N'-(3-methoxybenzylidene)hydraz-
ino]isoquinolin-4-yl}methoxy-benzamide, Q-1, as a yellow solid (25
mg, 21%): mp 125-129.degree. C.; HPLC R.sub.t=18.3 min.; TLC
R.sub.f=0.5 (4% methanol/dichloromethane); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.94 (s, 1H), 8.43 (br s, 2H), 7.65-7.22 (m,
13H), 7.20 (d, 1H, J=8.4 Hz), 6.99 (d, 1H, J=8.2 Hz), 5.24 (s, 2H),
3.84 (s, 3H), 3.10-3.05 (m, 1H), 2.29 (s, 3H), 1.17 (d, 6H, J=6.8
Hz); MS (ESI) m/z 559 [M+H].sup.+. Anal. calc'd for
C.sub.35H.sub.34N.sub.4O.sub.3.0.5 hexane: C, 75.84; H, 6.87; N,
9.31. Found: C, 75.81; H, 6.89; N, 9.09.
EXAMPLE R-1
N-(3,5-Diallyl-4-methyl-phenyl)-3-(isoquinolin-4-ylmethoxy)-benzamide
[0378] 188
[0379] (a) To a solution of 1.0 g (4.7 mmol) of
4-(chloromethyl)isoquinoli- ne hydrochloride, K-1c, in DMF (10 mL)
and acetone (25 mL) was added methyl 3-hydroxybenzoate (800 mg, 5.3
mmol) and cesium carbonate (3.8 g, 11.7 mmol). After refluxing for
20 h the solution was cooled, concentrated and diluted with
dichloromethane. The solution was washed with sodium bicarbonate
and brine. After drying over the sodium sulfate the product was
precipitated from diethyl ether/hexanes yielding 1.15 grams (84%
yield) of methyl 3-(isoquinolin-4-yl)methoxy-benzoate, R-1a, as a
white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 9.35 (s,
1H), 8.67 (s, 1H), 8.20 (d, J=8.0 Hz, 1H), 8.15 (d, J=8.6 Hz, 1H),
7.88 (ddd, J=7.0, 7.0, 1.3 Hz, 1H), 7.75 (ddd, J=7.5, 7.5, 1.0 Hz,
1H), 7.65 (m, 1H), 7.60 (m, 1H), 7.49 (dd, J=8.1 Hz, 1H), 7.41 (m,
1H), 5.64 (s, 2H), 3.86 (s, 3H).
[0380] (b) To methyl 3-(isoquinolin-4-yl)methoxy-benzoate, R-1a,
(2.24 g, 7.64 mmol) in 95% EtOH (50 mL) was added 1 N NaOH (8 ml).
After stirring for two hours at 70.degree. C. the solution was
cooled and concentrated to about 10 mL. With ice cooling the
solution was acidified with 1N HCl. The resulting white precipitate
was filtered yielding 1.3 g of 3-(isoquinolin-4-yl)methoxy-benzoic
acid hydrochloride, R-1b, as a white solid. Extraction of the
mother liquors with dichloromethane three times resulted in 480 mg
more product. (74% overall yield). .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 9.77 (s, 1H), 8.81 (s, 1H), 8.50 (d, J=8.2
Hz, 1H), 8.39 (d, J=8.5 Hz, 1H), 8.18 (dd, J=8.4, 8.4 Hz, 1H), 7.99
(dd, J=7.5, 7.5 Hz, 1H), 7.69 (m, 1H), 7.61 (m, 1H), 7.48 (dd,
J=8.1, 8.1 Hz, 1H), 7.41 (m, 1H), 5.76 (s, 2H).
[0381] (c) To a solution of 72 mg (0.38 mmol)
3,5-diallyl-4-methylaniline, prepared as described in steps (d)
through (f) below, in DMF (6 mL) was added 120 mg (0.380 mmol) of
3-(isoquinolin-4-ylmethoxy)-benzoic acid hydrochloride, R-1b, 160
.mu.L (0.92 mmol) of diisopropylethylamine and PyBop,(240 mg, 0.46
mmol). After stirring for 2.5 h, DMAP (3 mg) was added. At five
hours all but 3 mL of solvent was removed, and the remaining
solution was diluted with dichloromethane (40 ml) and washed with
water, and brine. After drying over sodium sulfate, the solution
was concentrated and chromatographed with silica gel eluting with
20 to 40% ethyl acetate/dichloromethane yielding 130 mg (75% yield)
of
N-(3,5-diallyl-4-methyl-phenyl)-3-(3soquinolin-4-yl)methoxy-benzamide,
R-1, as a white foam. .sup.1H NMR.(300 MHz, CDCl.sub.3) .delta.
9.29 (s, 1H), 8.63 (s, 1H), 8.06 (m, 2H), 7.79 (m, 1H), 7.62-7.82
(m, 3H), 7.44 (m, 2H), 7.35 (s, 2H), 7.23 (m, 1H), 5.97 (m, 2H),
5.53 (s, 2H), 5.03 (m, 4H), 3.41 (d, 4H, J=6.2 Hz), 2.18 (s, 3H).
MS (FAB) m/z 449 [M+H].sup.+. Anal. calcd for
C.sub.30H.sub.28N.sub.2O.sub.2.0.35H.sub.2O: C, 79.22; H, 6.36; N,
6.16. Found: C, 79.14; H, 6.24; N, 6.43.
[0382] (d) To 3,5-dibromoaniline (2.0 g, 6.5 mmol) in
CH.sub.2Cl.sub.2 (40 mL) was added diisopropylethylamine (2.27 mL,
13 mmol). The solution was cooled to 0.degree. C. followed by
addition of trifluoroacetic anhydride (1.37 mL, 9.8 mmol) in
CH.sub.2Cl.sub.2 (5 mL) over three minutes. After addition the
cooling bath was removed, and DMAP (approximately 3 mgs) was added.
At one hour the solution was diluted with CH.sub.2Cl.sub.2 (40 mL)
and washed with water and brine and dried over sodium sulfate. The
solution was concentrated and chromatographed though a short plug
of silica gel eluting with 30% ethyl acetate/hexanes which resulted
in 2.68 g, (quantitative yield), of
N-(3,5-dibromo-4-methyl-phenyl)-2,2,2-trifluo- ro-acetamide an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.80 (s,
3H, including NH), 2.56 (s, 3H).
[0383] (e) To dry, degassed toluene (25 mL) was added
N-(3,5-dibromo-4-methyl-phenyl)-2,2,2-trifluoroacetamide (1.0 g,
2.78 mmol), allyltributyltin (2.6 mL, 8.4 mmol) and
Pd(PPh.sub.3).sub.4 (200 mgs, 0.160 mmol). After refluxing for 14
hours, most of the solvent was removed and the solution was diluted
with diethyl ether. Approximately 100 mL of water was added
followed by DBU (1.4 mL, 0.92 mmol) which resulted in a gummy
precipitate. This heterogeneous solution was filtered through a
short plug of silica gel eluting with diethyl ether. After
concentration the residue was chromotographed twice with 10 to 20%
ethyl acetate/hexanes resulting in 315 mg (40% yield) of
N-(3,5-diallyl-4-methyl-phenyl)-2,2,2-trifluoroacetamide as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.72 (br s, 1H),
7.25 (s, 2H), 5.94 (m, 2H), 5.09 (dd, J=10.2, 1.5 Hz, 2H), 4.98
(dd, J=15.4, 1.8 Hz), 3.40 (d, J=6.2 Hz), 2.18 (s, 3H).
[0384] (f) To
N-(3,5-diallyl-4-methyl-phenyl)-2,2,2-trifluoroacetamide (255 mg,
0.90 mmol) in 95% ethanol (10 mL) was added 1N NaOH.sub.(aq) (2
mL). After heating to 80.degree. C. for 16 h, the solution was
cooled to room temperature. After concentration the residue was
diluted with ethyl acetate (30 mL) and washed with water and brine
and dried over sodium sulfate. Removal of the solvent led to 160 mg
(95% yield) of 3,5-diallyl-4-methylaniline as an orange oil.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.42 (s, 2H), 5.91 (m,
2H), 5.04 (dd, J=10.6, 1.7 Hz, 2H), 4.98 (dd, J=17.2. 1.8 Hz), 3.31
(d, J=6.3 Hz), 2.08 (s, 3H).
EXAMPLE R-2
N-(3,5-Dibromo-4-methyl-phenyl)-3-(isoquinolin-4-ylmethoxy)-benzamide
[0385] 189
[0386] Example R-2 was prepared in a similar manner to that
described for R-1, except that 3,5-dibromo-4-methylaniline was used
in place of 3,5-diallyl-4-bromoaniline in step (c): .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 10.36 (s, 1H), 9.36 (s, 1H), 8.69
(s, 1H), 8.20 (m, 1H), 8.14 (s, 2H), 7.89 (m, 1H), 7.76 (m, 1H),
7.68 (m, 1H), 7.58 (m, 1H), 7.51 (m, 1H), 7.38 (m, 1H), 5.65 (s,
2H), 2.48 (s, 3H). Anal. calc'd for
C.sub.24H.sub.18N.sub.2O.sub.2Br.sub.2.0.4H.sub.2O: C, 54.04; H,
3.55; N, 5.25. Found: C, 53.96; H, 3.50; N, 5.08.
EXAMPLE R-3
3-(Isoquinolin-4-ylmethoxy)-N-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-naph-
thalen-2-yl)-benzamide
[0387] 190
[0388] Example R-3 was prepared in a similar manner to that
described for R-1, except that
2-amino-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-naphthale- ne was
used in place of 3,5-diallyl-4-bromoaniline in step (c): .sup.1H
NMR (300 MHz, DMSO-d.sub.6) .delta. 10.12 (s, 1H), 9.77 (s, 1H),
8.23 (s, 1H), 8.49 (d, 1H, J=8.2 Hz), 8.39 (d, 1H, J=8.4 Hz), 8.18
(dd, 1H, J=7.8 Hz, J=7.7 Hz), 7.99 (dd, 1H, J=7.4 Hz, J=7.4 Hz),
7.76 (s, 1H), 7.70 (d, 1H, J=2.2 Hz), 7.63 (d, 1H, J=7.7 Hz), 7.58
(dd, 1H, J=8.6 Hz, J=2.2 Hz), 7.51 (dd, 1H, J=7.7 Hz, J=7.7 Hz),
7.38 (dd, 1H, J=8.2 Hz, J=1.9 Hz), 7.28, (d, 1H, 8.6 Hz), 5.79 (s,
2H), 1.64 (s, 4H), 1.25 (s, 6H), 1.24 (s, 6H). MS (ESI) m/z 465
[M+H].sup.+. Anal. calc'd for C.sub.31H.sub.32N.sub.2O.sub.2.1.0
HCl 0.8H.sub.2O: C, 72.23; H, 6.77; N, 5.43. Found: C, 72.07; H,
6.63; N, 5.43.
[0389] The intermediate
2-amino-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-nap- hthalene was
prepared by reduction of 2-nitro-5,5,8,8-tetramethyl-5,6,7,8--
tetrahydro-naphthalene (Kagechika, H. et al., J. Med. Chem., 31,
2182-2192 (1998)) in a similar manner to that described in step (b)
of Example J-1.
EXAMPLE R-4
3-(Isoquinolin-4-ylmethoxy)-N-(3-trifluoromethoxy-phenyl)-benzamide
[0390] 191
[0391] Example R-4 was prepared in a similar manner to that
described for R-1, except that 3-(trifluoromethoxy)aniline was used
in place of 3,5-diallyl-4-bromoaniline in step (c): .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.22 (s, 1H), 8.55 (s, 1H), 7.99 (m,
2H), 7.73 (m, 2H), 7.65 (m, 1H), 7.60 (m, 1H), 7.54 (m, 1H), 7.47
(m, 1H), 7.27-7.40 (m, 2H), 7.18 (m, 1H), 6.99 (m, 1H), 5.41 (s,
2H). MS (ESI) m/z 439 [M+H].sup.+. Anal. calc'd for
C.sub.24H.sub.17F.sub.3 N.sub.2O.sub.3: C, 65.75; H, 3.91; N, 6.39.
Found: C, 65.58; H, 4.02; N, 6.37.
EXAMPLE R-5
N-(2,4-Dimethylquinolin-6-yl)-3-(isoquinolin-4-ylmethoxy)-benzamide
[0392] 192
[0393] Example R-5 was prepared in a similar manner to that
described for R-1, except that 6-amino-2,4-dimethylquinoline was
used in place of 3,5-diallyl-4-bromoaniline in step (c): .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 10.50 (s, 1H), 9.36 (s, 1H), 8.70
(s, 1H), 8.56 (d, 1H, J=2.2 Hz), 8.20 (m, 2H), 8.06 (m, 1H), 7.9
(m, 2H), 7.76 (m, 2H), 7.66 (m, 1H), 7.53 (dd, 1H, J=7.8, 7.8 Hz),
7.39 (m, 1H), 7.27 (s, 1H), 5.67 (s, 2H), 2.61 (s, 3H), 2.58 (s,
3H). MS (FAB) m/z 435 [M+H].sup.+. Anal. calcd for
C.sub.27H.sub.22N.sub.4O.sub.2.0.1H.sub.2O: C, 74.33; H, 5.13; N,
12.84. Found: C, 79.13; H, 4.97; N, 12.74.
[0394] The intermediate 6-amino-2,4-dimethylquinoline was prepared
by reduction of 6-nitro-2,4-dimethylquinoline (Price, C. et al., J.
Org. Chem., 12, 203 (1947)) in a similar manner to that described
in step (b) of Example J-1.
EXAMPLE R-6
3-(Isoquinolin-4-ylmethoxy)-benzoic acid
N'-(4-trifluoromethyl-phenyl)-hyd- razide
[0395] 193
[0396] Example R-6 was prepared in a similar manner to that
described for R-1, except that 4-trifluoromethylphenylhydrazine was
used in place of 3,5-diallyl-4-bromoaniline in step (c): .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 9.28 (s, 1H), 8.62 (s, 1H), 8.05
(m, 3H), 7.79(dd, 1H, J=8.36 Hz, J=8.27 Hz), 7.67 (dd, 1H, J=7.98
Hz, J=8.14 Hz), 7.59 (m, 1H), 7.51-7.42 (m, 4H), 7.27 (m, 1H), 6.98
(s, 1H), 6.95 (s, 1H), 6.47 (br, 1H), 5.51 (s, 2H). MS (ESI) m/z
438 [M+H].sup.+. Anal. calc'd for
C.sub.24H.sub.18F.sub.3N.sub.3O.sub.2: C, 65.90; H, 4.15; N, 9.61.
Found: C, 65.75; H, 4.20; N, 9.51.
EXAMPLE R-7
N-Benzyloxy-3-(isoquinolin-4-ylmethoxy)-benzamide
[0397] 194
[0398] Example R-2 was prepared in a similar manner to that
described for R-1, except that O-benzylhydroxylamine was used in
place of 3,5-diallyl-4-bromoaniline in step (c): .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 9.26 (s, 1H), 8.59 (s, 1H), 8.57 (s, 1H),
8.05 (m, 2H), 7.78 (dd, 1H, J=8.23 Hz, J=8.44 Hz), 7.67 (dd, 1H,
J=7.89 Hz, J=8.16 Hz), 7.47-7.43 (m, 3H), 7.42-7.32 (m, 3H),
7.24-7.16 (m, 3H), 5.47 (s, 2H), 5.05 (s, 2H). MS (ESI) r/z 385
[M+H].sup.+. Anal. calc'd for C.sub.24H.sub.20N.sub.2O.sub.3: C,
74.98; H, 5.24; N, 7.29. Found: C, 74.85; H, 5.31; N, 7.18.
EXAMPLE R-8
3-(Isoquinolin-4-ylmethoxy)-Benzoic Acid N'-phenyl-hydrazide
[0399] 195
[0400] Example R-2 was prepared in a similar manner to that
described for R-1, except that phenyl-hydrazine was used in place
of 3,5-diallyl-4-bromoaniline in step (c): .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 9.28 (s, 1H), 8.62 (s, 1H), 8.06 (s, 1H), 8.04
(s, 1H), 7.94 (m, 1H), 7.78 (m, 1H), 7.67 (m, 1H), 7.59 (m, 1H),
7.44 (m, 2H), 7.25 (m, 3H), 6.93 (m, 3H), 6.38 (m, 1H), 5.50 (s,
2H). MS (ESI) m/z 370 [M+H].sup.+. Anal. calcd for C.sub.23H.sub.19
N.sub.3O.sub.2.0.35H.sub.2O- : C, 73.52; H, 5.29; N, 11.18. Found:
C, 73.72; H, 5.36; N, 10.90.
EXAMPLE R-9
N-(5,7-dimethyl[1,8]naphthydrin-2-yl)-3-(isoquinolin-4-ylmethoxy)-benzamid-
e
[0401] 196
[0402] Example R-9 was prepared in a similar manner to that
described for R-1, except that
2-amino-5,7-dimethyl[1,8]naphthydrine was used in place of
3,5-diallyl-4-bromoaniline in step (c): .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 9.30 (s, 1H), 8.96 (s, 1H), 8.64 (s, 2H), 8.40
(d, 1H, J=9.0 Hz), 8.08 (m, 2H), 7.81 (m, 1H), 7.73 (m, 1H), 7.69
(m, 1H), 7.62 (m, 1H), 7.48 (dd, 1H, J=7.8, 7.8 Hz), 7.29 (m, 1H),
7.15 (d, J=0.7 Hz), 5.55 (s, 2H), 2.72 (s, 3H), 2.69 (d, 3H, J=0.7
Hz). MS (FAB) m/z 435 [M+H].sup.+. Anal. calc'd for
C.sub.27H.sub.22N.sub.4O.sub.2.0.1H.sub.2O: C, 74.33; H, 5.13; N,
12.84. Found: C, 79.13; H, 4.97; N, 12.74
EXAMPLE R-10
3-(Isoquinolin-4-ylmethoxy)-N-(1,1,3,3-tetramethyl-1,3-dihydroisobenzofura-
n-5-yl)-benzamide
[0403] 197
[0404] Example R-10 was prepared in a similar manner to that
described for R-1, except that
5-amino-1,1,3,3-tetramethyl-1,3-dihydroisobenzofuran was used in
place of 3,5-diallyl-4-bromoaniline in step (c): .delta. .sup.1H
NMR (300 MHz, CDCl.sub.3 9.29 (s, 1H), 8.63 (s, 1H), 8.06 (m, 2H),
7.90 (m, 1H), 7.79 (dd, 1H, J=8.38 Hz, J=7.72 Hz), 7.68 (dd, 1H,
J=8.05 Hz, J=8.08 Hz), 7.62 (m, 1H), 7.55 (d, 1H, J=1.8 Hz), 7.45
(m, 2H), 7.40 (m, 1H), 7.22 (m, 1H), 7.08 (d, 1H, J=8.12 Hz), 5.52
(s, 2H), 1.54 (s, 6H), 1.51 (s, 6H). MS (ESI) m/z 453 [M+H].sup.+.
Anal. calcd for C.sub.29H.sub.28N.sub.2O.sub.3.times.0.4H.sub.2O:
C, 75.76; H, 6.31; N, 6.09. Found: C, 75.72; H, 6.31; N, 5.94.
EXAMPLE R-11
N-(3,5-Dichloro-4-pyrrolidin-1-yl-phenyl)-4-fluoro-3-(pyridin-3-ylmethoxy)-
-benzamide
[0405] 198
[0406] Example R-11 was prepared in a similar manner to that
described for R-1, except that ethyl 4-fluoro-3-hydroxybenzoate,
prepared by conventional Fischer esterification of
4-fluoro-3-hydroxybenzoic acid, was used in place of methyl
3-hydroxybenzoate and 3-picolyl chloride hydrochloride was used in
place of 4-(chloromethyl)isoquinoline hydrochloride, K-1c, in step
(a), and 1-(4-amino-2,6-dichlorophenyl)pyrro- lidine was used in
place of 3,5-diallyl-4-bromoaniline in step (c): mp 163-167.degree.
C.; HPLC R.sub.t=17.7 min.; TLC R.sub.f=0.3 (40% ethyl
acetate/cyclohexane); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
10.40 (s, 1H), 8.72 (s, 1H), 8.60 (d, 1H, J=3.9 Hz), 7.94-7.84 (m,
4H), 7.66-7.62 (m, 1H), 7.49-7.42 (m, 2H), 5.33 (s, 2H), 3.24-3.20
(m, 4H), 1.98-1.96 (m, 4H); MS (ESI) m/z 460 [M+H].sup.+. Anal.
calc'd for C.sub.23H.sub.20Cl.sub.2FN.sub.3O.sub.2: C, 60.01; H,
4.38; N, 9.13. Found: C, 60.08; H, 4.49; N, 9.02.
EXAMPLE R-12
4-Fluoro-N-(4-morpholin-4-yl-3-trifluoromethyl-phenyl)-3-(pyridin-3-ylmeth-
oxy)-benzamide
[0407] 199
[0408] Example R-12 was prepared in a similar manner to that
described for R-1, except that ethyl 4-fluoro-3-hydroxybenzoate,
prepared by conventional Fischer esterification of
4-fluoro-3-hydroxybenzoic acid, was used in place of methyl
3-hydroxybenzoate and 3-picolyl chloride hydrochloride was used in
place of 4-(chloromethyl)isoquinoline hydrochloride, K-1c, in step
(a), and 1-(4-amino-2-trifluoromethylphenyl)- morpholine was used
in place of 3,5-diallyl-4-bromoaniline in step (c): mp
160-161.degree. C.; HPLC R.sub.t=14.4 min.; TLC R.sub.f=0.2 (50%
ethyl acetate/cyclohexane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.46 (s, 1H), 8.72 (s, 1H), 8.60 (d, 1H, J=4.1 Hz), 8.14
(d, 1H, J=2.4 Hz), 8.06 (dd, 1H, J=8.8, 2.4 Hz), 7.94-7.86 (m, 2H),
7.69-7.61 (m, 2H), 7.50-7.41 (m, 2H), 5.34 (s, 2H), 3.71 (t, 4H,
J=4.4 Hz), 2.85 (t, 4H, J=4.4 Hz); MS (ESI) m/z 476 [M+H].sup.+.
Anal. calc'd for C.sub.24H.sub.21F.sub.4N.sub.3O.sub.3: C, 60.63;
H, 4.45; N, 8.84. Found: C, 60.84; H, 4.57; N, 8.81.
EXAMPLE R-13
4-Fluoro-N-[4-(piperazin-1-yl)-3-trifluoromethylphenyl]-3-(pyridin-3-yl)Me-
thoxybenzamide
[0409] 200
[0410] (a)
4-Fluoro-N-[4-{4-(t-butoxycarbonyl)piperazin-1-yl}-3-trifluorom-
ethylphenyl]-3-(pyridin-3-yl)methoxybenzamide, R-13a, was prepared
in a similar manner to that described for Example R-1, except that
ethyl 4-fluoro-3-hydroxybenzoate, prepared by conventional Fischer
esterification of 4-fluoro-3-hydroxybenzoic acid, was used in place
of methyl 3-hydroxybenzoate and 3-picolyl chloride hydrochloride
was used in place of 4-(chloromethyl)isoquinoline hydrochloride,
K-.sub.1c, in step (a), and
1-(4-amino-2-trifluoromethylphenyl)-4-(t-butoxycarbonyl)piperazi-
ne was used in place of 3,5-diallyl-4-bromoaniline in step (c).
[0411] (b) A solution of
4-fluoro-N-[4-{4-(t-butoxycarbonyl)piperazin-1-yl-
}-3-trifluoromethylphenyl]-3-(pyridin-3-yl)methoxybenzamide, R-13a,
(65 mg, 0.11 mmol) in methylene chloride (4 mL) was treated with
trifluoroacetic acid (1 mL). After 18 hours, the solution was
concentrated under reduced pressure. The resultant residue was
treated with 5% sodium bicarbonate (25 mL) and extracted with ethyl
acetate (3.times.25 mL). The combined organic extracts were washed
with water (50 mL), brine (50 mL), dried over magnesium sulfate,
filtered, and concentrated under reduced pressure to give a tan
solid (48 mg). The crude product was purified by radial
chromatography over silica gel using 5-15% methanol/chloroform with
0.1% ammonium hydroxide to give
4-fluoro-N-[4-(piperazin-1-yl)-3-trifluoromethylphenyl]-3-(pyridin-3-yl)m-
ethoxybenzamide, R-13, as a white solid (34 mg, 63%): mp
123-131.degree. C.; HPLC R.sub.t=13.2 min.; TLC R.sub.f=0.3 (6%
methanol/chloroform w/ 0.1% NH.sub.4OH); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.43 (s, 1H), 8.72 (d, 1H, J=1.6 Hz), 8.60
(dd, 1H, J=4.8, 1.5 Hz), 8.12 (d, 1H, J=2.3 Hz), 8.05-8.02 (m, 1H),
7.94-7.86 (m, 2H), 7.69-7.64 (m, 1H), 7.54 (d, 1H, J=8.8 Hz),
7.49-7.41 (m, 2H), 5.33 (s, 2H), 2.82-2.79 (m, 4H), 2.78-2.75 (m,
4H); MS (ESDI) m/z 475 [M+H].sup.+. Anal. calc'd for
C.sub.24H.sub.22F.sub.4N.sub.4O.sub.2: C, 60.76; H, 4.67; N, 11.81.
Found: C, 60.66; H, 4.98; N, 11.38.
EXAMPLE R-14
4-Fluoro-N-(4-morpholin-4-yl-3-trifluoromethyl-phenyl)-3-(isoquinolin-4-yl-
methoxy)-benzamide
[0412] 201
[0413] Example R-14 was prepared in a similar manner to that
described for R-1, except that ethyl 4-fluoro-3-hydroxybenzoate,
prepared by conventional Fischer esterification of
4-fluoro-3-hydroxybenzoic acid, was used in place of methyl
3-hydroxybenzoate in step (a), and
1-(4-amino-2-trifluoromethylphenyl)morpholine was used in place of
3,5-diallyl-4-bromoaniline in step (c): mp 175-180.degree. C.; HPLC
R.sub.t=15.3 min.; TLC R.sub.f=0.3 (1% methanol/methylene
chloride); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.67 (s,
1H), 9.80 (s, 1H), 8.83 (s, 1H), 8.52 (d, 1H, J=8.2 Hz), 8.44 (d,
1H, J=8.5 Hz), 8.23-8.17 (m, 3H), 8.12 (dd, 1H, J=8.8, 2.3 Hz),
8.00 (t, 1H, J=7.4 Hz), 7.74-7.70 (m, 1H), 7.62 (d, 1H, J=8.7 Hz),
7.46 (dd, 1H, J=11.0, 8.4 Hz), 5.91 (s, 2H), 3.71 (t, 4H, J=4.2
Hz), 2.84 (t, 4H, J=4.2 Hz); MS (ESI) m/z 526 [M+H].sup.+.
[0414] Anal. calcd for C.sub.28H.sub.23F.sub.4N.sub.3O.sub.3.1.3
HCl: C, 58.70; H, 4.28; N, 7.33; Cl, 8.05. Found: C, 59.05; H,
4.59; N, 7.28; Cl, 8.01.
EXAMPLE R-15
4-Fluoro-N-(4-piperazin-1-yl-3-trifuoromethyl-phenyl)-3-(isoquinolin-4-ylm-
ethoxy)-benzamide
[0415] 202
[0416] Example R-15 was prepared in a similar manner to that
described for R-13, except that 4-(chloromethyl)isoquinoline
hydrochloride, K-1c, was used in place of 3-picolyl chloride
hydrochloride in step (a): mp 103-107.degree. C.; HPLC R.sub.t=14.7
min.; TLC R.sub.f=0.3 (5% methanol/chloroform with 0.1%
NH.sub.4OH); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.45 (s,
1H), 9.37 (s, 1H), 8.68 (s, 1H), 8.23-8.17 (m, 2H), 8.14-8.13 (m,
1H), 8.06-8.02 (m, 2H), 7.92-7.87 (m, 1H), 7.79-7.74 (M, 1H),
7.69-7.65 (m, 1H), 7.55 (d, 1H, J=8.5 Hz), 7.42 (dd, 1H, J=11.0,
8.6 Hz), 5.72 (s, 2H), 2.80 (br. s, 4H), 2.77 (br. s, 4H); MS (ESI)
m/z 525 [M+H].sup.+. Anal. calcd for
C.sub.28H.sub.24F.sub.4N.sub.4O.0.1 hexanes (MW 533.1 g/mol): C,
64.43; H, 4.80; N, 10.51. Found: C, 64.68; H, 5.07; N, 10.16.
EXAMPLE R-16
4-Fluoro-N-(4-morpholin-4-yl-3-trifluoromethyl-phenyl)-3-(quinolin-3-ylmet-
hoxy)-benzamide
[0417] 203
[0418] Example R-16 was prepared in a similar manner to that
described for R-1, except that ethyl 4-fluoro-3-hydroxybenzoate,
prepared by conventional Fischer esterification of
4-fluoro-3-hydroxybenzoic acid, was used in place of methyl
3-hydroxybenzoate and 3-chloromethylquinoline hydrochloride was
used in place of 4-(chloromethyl)isoquinoline hydrochloride, K-1c,
in step (a), and 1-(4-amino-2-trifluoromethylphenyl)- morpholine
was used in place of 3,5-diallyl-4-bromoaniline in step (c): mp
81-84.degree. C.; HPLC R.sub.t=15.4 min.; TLC R.sub.f=0.5 (1%
methanol/methylene chloride); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.46 (s, 1H), 9.04 (d, 1H, J=2.2 Hz), 8.49 (s, 1H), 8.14
(d, 1H, J=2.4 Hz), 8.08-8.04 (m, 3H), 7.93 (dd, 1H, J=8.2, 1.8 Hz),
7.83-7.78 (m, 1H), 7.70-7.60 (m, 3H), 7.46 (dd, 1H, J=11.0, 8.5
Hz), 5.52 (s, 2H), 3.70 (t, 4H, J=4.3 Hz), 2.84 (t, 4H, J=4.3 Hz);
MS (ESI) m/z 526 [M+H].sup.+. Anal. calc'd for
C.sub.28H.sub.23F.sub.4N.sub.3O.sub.3: C, 64.00; H, 4.41; N, 8.00.
Found: C, 64.15; H, 4.53; N, 7.97.
EXAMPLE R-17
4-Fluoro-N-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-3-(quinolin-3-ylmet-
hoxy)-benzamide
[0419] 204
[0420] Example R-17 was prepared in a similar manner to that
described for R-13, except that 3-(chloromethyl)quinoline
hydrochloride was used in place of 3-picolyl chloride hydrochloride
in step (a): mp 76-78.degree. C.; HPLC R.sub.t=14.7 min.; TLC
R.sub.f=0.3 (3% methanol/chloroform w/ 0.1% NH.sub.4DH); .sup.1H
NMR (300 MHz, DMSO-d.sub.6) .delta. 10.44 (s, 1H), 9.04 (d, 1H,
J=2.0 Hz), 8.48 (s, 1H), 8.12 (d, 1H, J=2.1 Hz), 8.08-8.02 (m, 3H),
7.93-7.91 (1,1H), 7.83-7.78 (m, 1H), 7.68-7.63 (m, 2H), 7.54 (d;
1H, J=8.9 Hz), 7.46 (dd, 1H, J=11.0, 8.5 Hz), 5.52 (s, 2H), 2.81
(br. s, 4H), 2.78 (br. s, 4H); MS (ESI) m/z 525 [M+H].sup.+. Anal.
calc'd for C.sub.28H.sub.24F.sub.4N.sub.4O.sub.2: C, 64.12; H,
4.61; N, 10.68. Found: C, 64.53; H, 4.99; N, 10.25.
EXAMPLE R-18
N-(3,5-Dichloro-4-morpholin-4-yl-phenyl)-4-fluoro-3-(pyridin-3-ylmethoxy)--
benzamide
[0421] 205
[0422] Example R-18 was prepared in a similar manner to that
described for R-1, except that ethyl 4-fluoro-3-hydroxybenzoate,
prepared by conventional Fischer esterification of
4-fluoro-3-hydroxybenzoic acid, was used in place of methyl
3-hydroxybenzoate and 3-picolyl chloride hydrochloride was used in
place of 4-(chloromethyl)isoquinoline hydrochloride, K-1c, in step
(a), and 1-(4-amino-2,6-dichlorophenyl)morph- oline was used in
place of 3,5-diallyl-4-bromoaniline in step (c): mp 209-211.degree.
C.; HPLC R.sub.t=15.0 min.; TLC R.sub.f=0.3 (40% ethyl
acetate/cyclohexane); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
10.41 (s, 1H), 8.71 (s, 1H), 8.58 (d, 1H, J=4.7 Hz), 7.93-7.82 (m,
4H), 7.63-7.60 (m, 1H), 7.49-7.41 (m, 2H), 5.32 (s, 2H), 3.70 (br.
s, 4H), 3.12 (br. s, 4H); .sup.13C NMR (75 MHz, DMSO-d.sub.6)
.delta. 164.4, 154.0 (d, J.sub.CF=250 Hz), 149.4, 149.2, 146.0 (d,
J.sub.CF=11 Hz), 139.5, 137.4, 136.0, 134.2, 131.8, 130.9 (d,
J.sub.CF=3 Hz), 123.8, 121.6 (d, J.sub.CF=8 Hz), 120.4, 116.1 (d,
J.sub.CF=19 Hz), 115.0 (d, J.sub.CF=2 Hz), 68.3, 67.0, 49.4; MS
(ESI) m/z 476 [M+H].sup.+. Anal. calc'd for
C.sub.23H.sub.20Cl.sub.2FN.sub.3O.sub.3: C, 58.00; H, 4.23; Cl,
8.82; N, 14.89. Found: C, 57.89; H, 4.24; Cl, 14.88; N, 8.69.
EXAMPLE R-19
N-(3,5-Dichloro-4-piperazin-1-yl-phenyl)-4-fluoro-3-(pyridin-3-ylmethoxy)--
benzamide
[0423] 206
[0424] Example R-19 was prepared in a similar manner to that
described for R-13, except that
1-(4-amino-2,6-dichlorophenyl)-4-(t-butoxycarbonyl)pipe- razine was
used in place of 1-(4-amino-2-trifluoromethylphenyl)-4-(t-butox-
ycarbonyl)piperazine in step (a) mp 81-85.degree. C.; HPLC
R.sub.t=11.7 min.; TLC R.sub.f=0.4 (5% methanol/chloroform w/ 0.1%
NH.sub.4OH); .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.38 (s,
1H), 8.71 (d, 1H, J=1.8 Hz), 8.58 (dd, 1H, J=4.7, 1.3 Hz),
7.93-7.82 (m, 4H), 7.65-7.60 (m, 1H), 7.49-7.41 (m, 2H), 5.32 (s,
2H), 3.05-3.02 (m, 4H), 2.88-2.79 (m, 4H); .sup.3C NMR (75 MHz,
DMSO-d.sub.6) .delta. 164.7, 152.2 (d, J.sub.CF=250 Hz), 149.8,
149.5, 146.2 (d, J.sub.CF=11 Hz), 140.7, 137.3, 136.2, 134.4,
132.1, 131.2 (d, J.sub.CF=3 Hz), 124.0, 121.8 (d, J.sub.CF=8 Hz),
120.7, 116.4 (d, J.sub.CF=19 Hz), 115.3, 68.6, 50.8, 46.6; MS (ESI)
m/z 475 [M+H].sup.+. Anal. calc'd for
C.sub.23H.sub.21Cl.sub.2FN.sub.4O.sub.2: C, 58.12; H, 4.45; Cl,
14.92; N, 11.79. Found: C, 57.99; H, 4.60; Cl, 14.59; N, 11.38.
EXAMPLE R-20
4-Fluoro-N-[4-(piperazin-1-yl)-3-trifluoromethylphenyl]-3-(pyridin-3-yl)Me-
thoxybenzamide
[0425] 207
[0426] Example R-20 was prepared in a similar manner to that
described for R-1, except that ethyl 4-fluoro-3-hydroxybenzoate,
prepared by conventional Fischer esterification of
4-fluoro-3-hydroxybenzoic acid, was used in place of methyl
3-hydroxybenzoate and 3-picolyl chloride hydrochloride was used in
place of 4-(chloromethyl)isoquinoline hydrochloride, K-1c, in step
(a), and 1-(4-amino-2-trifluoromethylphenyl)- -4-methylpiperazine
was used in place of 3,5-diallyl-4-bromoaniline in step (c): mp
61-66.degree. C.; HPLC R.sub.t=13.0 min.; TLC R.sub.f=0.5 (5%
methanol/chloroform w/ 0.1% NH.sub.4OH); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.46 (s, 1H), 8.72 (s, 1H), 8.59 (d, 1H,
J=4.4 Hz), 8.14 (d, 1H, J=2.0 Hz), 8.05 (dd, 1H, J=8.6, 1.6 Hz),
7.94-7.86 (m, 2H), 7.69-7.65 (m, 1H), 7.60 (d, 1H, J=8.8 Hz),
7.49-7.41 (m, 2H), 5.33 (s, 2H), 2.92 (s, 4H), 2.88 (br. s, 4H),
2.42 (s, 3H); MS (ESI) m/z 489 [M+H].sup.+. Anal. calc'd for
C.sub.25H.sub.24F.sub.4N.sub.4O.sub.2 x 1.3H.sub.2O: C, 58.66; H,
5.24; N, 10.95. Found: C, 58.17; H, 4.80; N, 10.52.
EXAMPLE R-21
4-Fluoro-N-(4-imidazol-1-yl-3-trifluoromethyl-phenyl)-3-(pyridin-3-ylmetho-
xy)-benzamide
[0427] 208
[0428] Example R-21 was prepared in a similar manner to that
described for R-1, except that
imidazol-1-yl-trifluoromethyl-phenylamine, J-1b, was used in place
of 3,5-diallyl-4-bromoaniline and 4-fluoro-3-(pyridin-3-ylm-
ethoxy)-benzoic acid, which was prepared as described in R-11, was
used in place of 3-(isoquinolin-4-ylmethoxy)-benzoic acid, R-1b:
HPLC R.sub.t=12.8 min.; TLC R.sub.f=0.3 (5% methanol-chloroform w/
0.1% ammonium hydroxide); .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 10.73 (s, 1H), 8.73 (s, 1H), 8.59 (s, 1H), 8.39 (d, 1H,
J=2.2 Hz), 8.21 (dd, 1H, J=8.6, 2.1 Hz), 7.94-7.83 (m, 3H),
7.73-7.68 (m, 1H), 7.60 (d, 1H, J=8.7 Hz), 7.51-7.40 (m, 3H), 7.10
(s, 1H), 5.35 (s, 2H); MS (ESI) m/z 457 (M+H).sup.+. Anal. calcd
for C.sub.23H.sub.16F.sub.4N.sub.4O.sub.2: C, 60.53; H, 3.53; N,
12.28. Found: C, 60.37; H, 3.62; N, 12.21.
EXAMPLE R-22
4-Fluoro-N-(4-pyrazol-1-yl-3-trifluoromethyl-phenyl)-3-(pyridin-3-ylmethox-
y)-benzamide
[0429] 209
[0430] Example R-22 was prepared in a similar manner to that
described for R-1, except that
pyrazol-1-yl-trifluoromethyl-phenylamine was used in place of
3,5-diallyl-4-bromoaniline and 4-fluoro-3-(pyridin-3-ylmethoxy)--
benzoic acid, which was prepared as described in R-11, was used in
place of 3-(isoquinolin-4-ylmethoxy)-benzoic acid, R-1b: HPLC
R.sub.t 13.9 min.; TLC R.sub.f 0.2 (2% methanol-chloroform w/ 0.11%
ammonium hydroxide); .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta.
10.72 (s, 1H), 8.72 (d, 1H, J=1.8 Hz), 8.60 (dd, 1H, J=4.6, 1.2
Hz), 8.38 (d, 1H, J=2.3 Hz), 8.21 (dd, 1H, J=8.7, 2.3 Hz), 8.02 (d,
1H, J=2.2 Hz), 7.95-7.89 (m, 2H), 7.74-7.69 (m, 2H), 7.68 (d, 1H,
J=8.7 Hz), 7.51-7.44 (m, 2H), 6.52-6.51 (m, 1H), 5.35 (s, 2H); MS
(ESI) m/z 457 (M+H).sup.+. Anal. calcd for
C.sub.23H.sub.16F.sub.4N.sub.4O.sub.2: C, 60.53; H, 3.53; N, 12.28.
Found: C, 60.39; H, 3.64; N, 12.19.
EXAMPLE R-23
4-Fluoro-3-(pyridin-3-ylmethoxy)-N-(4-[1,2,4]triazol-1-yl-3-trifluoromethy-
l-phenyl)-benzamide
[0431] 210
[0432] Example R-23 was prepared in a similar manner to that
described for R-1, except that
[1,2,4]triazol-1-yl-trifluoromethyl-phenylamine was used in place
of 3,5-diallyl-4-bromoaniline and 4-fluoro-3-(pyridin-3-ylmethox-
y)-benzoic acid, which was prepared as described in R-11, was used
in place of 3-(isoquinolin-4-ylmethoxy)-benzoic acid, R-1b: HPLC
R.sub.t 12.4 min.; TLC R.sub.f 0.3 (5% methanol-chloroform w/ 0.1%
ammonium hydroxide); .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta.
10.79 (s, 1H), 8.89 (s, 1H), 8.73 (d, 1H, J=1.7 Hz), 8.60 (dd, 1H,
J=4.8, 1.8 Hz), 8.44 (d, 1H, J=2.3 Hz), 8.28-8.24 (m, 2H),
7.96-7.90 (m, 2H), 7.74-7.69 (m, 2H), 7.52-7.45 (m, 2H), 5.35 (s,
2H); MS (ESI) m/z 458 (M+H).sup.+. Anal. calcd for
C.sub.22H.sub.15F.sub.4N.sub.5O.sub.2: C, 56.65; H, 3.46; N, 15.02.
Found: C, 56.53; H, 3.44; N, 14.96.
EXAMPLE R-24
N-(3,5-Dichloro-4-imidazol-1-yl-phenyl)-4-fluoro-3-(pyridin-3-ylmethoxy)-b-
enzamide
[0433] 211
[0434] Example R-24 was prepared in a similar manner to that
described for R-1, except that
3,5-dichloro-4-imidazol-1-yl-phenylamine was used in place of
3,5-diallyl-4-bromoaniline and 4-fluoro-3-(pyridin-3-ylmethoxy)--
benzoic acid, which was prepared as described in R-11, was used in
place of 3-(isoquinolin-4-ylmethoxy)-benzoic acid, R-1b: HPLC
R.sub.t 13.0 min.; TLC R.sub.f 0.7 (5% methanol-dichloromethane);
.sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.65 (s, 1H), 8.89 (s,
1H), 8.72 (d, 1H, J=1.9 Hz), 8.60 (dd, 1H, J=4.8, 1.6 Hz), 8.11 (s,
2H), 7.95-7.91 (m, 1H), 7.88 (dd, 1H, J=8.2, 2.0 Hz), 7.81 (s, 1H),
7.71-7.66 (m, 1H), 7.51-7.44 (m, 2H), 7.33 (s, 1H), 7.13 (s, 1H),
5.34 (s, 2H); .sup.13C NMR (DMSO-d.sub.6, 75 MHz) .delta. 165.2,
154.6 (d, J.sub.CF=251.1 Hz), 149.9, 149.6, 146.4(d, J.sub.CF=11.1
Hz), 141.2, 138.4, 136.3, 133.1, 132.2, 130.9 (d, J.sub.CF=3.5 Hz),
129.2, 128.3, 124.1, 122.0 (d, J.sub.CF=7.6 Hz), 121.3, 119.9,
116.6 (d, J.sub.CF=18.7 Hz), 115.6 (d, J.sub.CF=1.7 Hz), 68.8; MS
(ESI) m/z 457 (M+H).sup.+. Anal. calcd for
C.sub.22H.sub.15Cl.sub.2FN.sub.4O.sub.2: C, 57.78; H, 3.31; N,
12.25; Cl, 15.51. Found: C, 57.38; H, 3.52; N, 11.90; Cl,
16.40.
EXAMPLE R-25
3-(5-Bromo-pyridin-3-ylmethoxy)-4-fluoro-N-(4-piperazin-1-yl-3-trifluorome-
thyl-phenyl)-benzamide
[0435] 212
[0436] (a) (5-Bromo-pyridin-3-yl)-methanol hydrochloride, R-25a,
was prepared according to the procedure described in J. Med. Chem.,
1997, 40, 2866-2875: HPLC R.sub.t 3.9 min.; TLC R.sub.f 0.2 (free
base; 40% ethyl acetate-cyclohexane); .sup.1H NMR (DMSO-d.sub.6,
300 MHz) .delta. 8.70 (d, 1H, J=2.2 Hz), 8.58-8.57 (m, 1H), 8.11
(t, 1H, J=2.0 Hz), 4.57 (s, 2H); .sup.13C NMR (DMSO-d.sub.6, 75
MHz) .delta. 144.1, 141.9, 140.2, 120.8, 59.9; MS (ESI) m/z 188/190
(M+H).sup.+.
[0437] (b) To a solution of (5-bromo-pyridin-3-yl)-methanol
hydrochloride, R-25a, (1.9 g, 8.5 mmol, 1.0 eq) was added thionyl
chloride (6 mL, 85 mmol, 10 eq). The amber solution was warmed to
70.degree. C. for 2 h. The crude product was cooled to room
temperature, diluted with toluene (50 mL) and concentrated under
reduced pressure to give 3-bromo-5-chloromethyl-pyridine
hydrochloride, R-25b, as a tan solid: HPLC R.sub.t 10.1 min.;
.sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 8.72 (d, 1H, J=2.2 Hz),
8.68 (d, 1H, J=1.7 Hz), 8.21 (t, 1H, J=2.0 Hz), 4.43 (s, 2H);
.sup.13C NMR (DMSO-d.sub.6, 75 MHz) .delta. 149.8, 148.0, 140.2,
136.4, 120.5, 42.3; MS (ESI) m/z 206/208 (M+H).sup.+.
[0438] (c)
4-[({1-[3-(5-Bromo-pyridin-3-ylmethoxy)-4-fluoro-phenyl]-methan-
oyl}-amino)-trifluoromethyl-phenyl]-piperazine-1-carboxylic acid
tert-butyl ester, R-25c, was prepared in a similar manner to that
described for R-1a, except that 3-bromo-5-chloromethyl-pyridine
hydrochloride, R-25b, was used in place of
4-(chloromethyl)isoquinoline hydrochloride, K-1c, and
4-({[1-(4-fluoro-3-hydroxy-phenyl)-methanoyl]-am-
ino}-trifluoromethyl-phenyl)-piperazine-1-carboxylic acid
tert-butyl ester, DD-1a, was used in place of 3-hydroxybenzoate:
HPLC R.sub.t 19.2 min.; TLC R.sub.f 0.3 (2%
methanol-dichloromethane); .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 10.44 (s, 1H), 8.74 (d, 1H, J=2.2 Hz), 8.72 (d, 1H, J=1.7
Hz), 8.21 (t, 1H, J=2.0 Hz), 8.14 (d, 1H, J=2.4 Hz), 8.04 (dd, 1H,
J=8.6, 2.4 Hz), 7.86 (dd, 1H, J=8.2, 2.0 Hz), 7.71-7.66 (m, 1H),
7.60 (d, 1H, J=8.8 Hz), 7.46 (dd, 1H, J=11.0, 8.5 Hz), 5.34 (s,
2H), 3.44 (br. s, 4H), 2.80 (t, 4H, J=4.7 Hz), 1.44 (s, 9H); MS
(ESI) m/z 653/655 (M+H).sup.+.
[0439] (d)
3-(5-Bromo-pyridin-3-ylmethoxy)-4-fluoro-N-(4-piperazin-1-yl-3--
trifluoromethyl-phenyl)-benzamide dihydrochloride, R-25, was
prepared in the manner similar to that described in example AA-1,
step (i), except
4-[({1-[3-(5-bromo-pyridin-3-ylmethoxy)-4-fluoro-phenyl]-methanoyl}-amino-
)-trifluoromethyl-phenyl]-piperazine-1-carboxylic acid tert-butyl
ester, R-25c, was used in place of
4-{[(1-{3-[2-(6-acetylamino-pyridin-3-yl)-eth-
yl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxy-
lic acid tert-butyl ester, AA-1 h: HPLC R.sub.t 13.1 min.; .sup.1H
NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.59 (s, 1H), 9.05 (br. s,
2H), 8.74-8.72 (m, 2H), 8.20 (d, 2H, J=1.9 Hz), 8.12 (d, 1H, J=8.7
Hz), 7.92 (d, 1H, 3=7.9 Hz), 7.71-7.68 (m, 1H), 7.56 (d, 1H, J=8.7
Hz), 7.44 (dd, 1H, J=11.0, 8.6 Hz), 5.36 (s, 2H), 3.17 (br. s, 4H),
3.07 (br. s, 4H); MS (ESI) m/z 553/555 (M+H).sup.+. Anal. calcd for
C.sub.24H.sub.21BrF.sub.4N- .sub.4O.sub.2x 2.0 HClx0.6H.sub.2O: C,
45.24; H, 3.83; N, 8.79; Br, 12.54; Cl, 11.13. Found: C, 45.20; H,
3.94; N, 8.50; Br, 12.18; Cl, 10.81.
EXAMPLE S-1
3-(2-Isoquinolin-4-yl-ethyl)-N-phenyl-benzamide
[0440] 213
[0441] (a) 4-Bromoisoquinoline (1.0 g, 4.8 mmol),
dichlorobis(triphenylpho- sphine)palladium (20.4 mg, 0.029 mmol),
copper iodide (1.5 mg, 0.008 mmol), trimethylsilylacetylene (707
mg, 7.2 mmol) and triethylamine (20 ml) were heated in a sealed
tube at 65.degree. C. for 16 h. After concentration the residue was
diluted with ethyl acetate, washed with brine, dried over sodium
sulfate, filtered and concentrated to dryness. The crude residue
was purified on silica gel using a gradient of 10% to 20% ethyl
acetate in hexanes as eluant to obtain 1.01 g (93%) of
4-(trimethyl-silanylethynyl)-isoquinoline, S-1a, as a yellow
liquid: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.18 (s, 1H),
8.70 (s, 1H), 8.34 (m, 1H), 8.14 (m, 1H), 7.78 (m, 1H), 7.65 (m,
1H), 0.35 (s, 9H).
[0442] (b) To a solution of NaOH (0.23 g, 5.78 mmol) in methanol
was added 1.0 g (4.44 mmol)
4-(trimethyl-silanylethynyl)-isoquinoline, S-1a. After stirring for
2 h at room temperature the methanol was removed followed by
addition of ethyl acetate. The organic solution was washed with
water, brine and dried over sodium sulfate. Removal of solvent led
to 0.65 g (96%) of 4-ethynyl-isoquinotine, S-1b: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 9.22 (s, 1H), 8.74 (s, 1H), 8.27 (m, 1H),
7.99 (m, 1H), 7.80 (m, 1H), 7.67 (m, 1H), 3.55 (s, 1H).
[0443] (c) 4-Ethynyl-isoquinoline, S-1b, (0.64 g, 4.18 mmol), ethyl
3-iodobenzoate (1.15 g, 4.18 mmol),
dichlorobis(triphenylphosphine)pallad- ium (7.02 mg, 0.010 mmol)
and copper iodide (0.4 mg, 0.002 mmol) in triethylamine (20 ml)
were stirred at room temperature for 14 h. The mixture was filtered
through celite, and the filtrate was concentrated to remove
triethylamine. To this residue was added ethyl acetate, and this
solution was washed with brine, dried over sodium sulfate, and
concentrated to dryness. The crude residue was purified on silica
gel using a gradient 0% to 5% ethyl acetate in dichloromethane as
eluant to obtain 1.1 g (87%) of ethyl
3-(isoquinolin-4-ylethynyl)benzoate, S-1c, as a semisolid: .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 9.23 (brs, 1H), 8.80 (br s, 1H),
8.32 (m, 2H), 8.07 (m, 1H), 8.03 (m, 1H), 7.84 (m, 2H), 7.69 (m,
2H), 7.50 (dd, 1H, J=7.4 Hz), 4.43 (q, 2H, J=7.1 Hz), 1.44 (t, 3H,
J=7.1 Hz).
[0444] (d) Ethyl 3-(isoquinolin-4-ylethynyl)benzoate, S-1c, (1.1 g,
3.64 mmol) and 10% Pd/C (0.5 g) in ethanol (20 ml) were stirred
under one atm of hydrogen at room temperature for 16 h. The
solution was filtered through celite, and the filtrate was
concentrated leaving 1.02 g (91.6%) of ethyl
3-(isoquinolin-4-ylethyl)benzoate, S-1d, as a liquid: .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.14 (s, 1H), 8.32 (s, 1H), 8.01 (m,
2H), 7.91 (m, 2H), 7.75 (m, 1H), 7.62 (m, 1H), 7.34 (m, 2H), 4.39
(q, 2H J=7.1 Hz), 3.35 (m, 2H), 3.10 (m, 2H), 1.40 (t, 3H, J=7.1
Hz).
[0445] (e) To a stirred solution of ethyl
3-(isoquinolin-4-ylethyl)benzoat- e, S-1d, (1.02 g, 3.3 mmol) in
methanol (20 ml) was added 1N NaOH (3.63 ml, 3.63 mmol). After
refluxing for 4 h, the methanol was removed. The solution was
diluted with water and upon acidification to pH 3 a white
precipitate formed which was subsequently filtered and dried under
high vacuum to obtain 0.92 g (100%) of
3-(2-isoquinolin-4-yl-ethyl)-benzoic acid, S-1e: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 9.75 (s, 1H), 8.54 (m, 3H), 8.21 (m, 1H),
8.02 (m, 1H), 7.89 (br s, 1H), 7.80 (m, 1H), 7.57 (m, 1H), 7.43
(dd, 1H, J=7.7 Hz), 3.52 (m, 2H), 3.11 (m, 2H).
[0446] (f) To a stirred solution of
3-(2-isoquinolin-4-yl-ethyl)-benzoic acid, S-1e, (0.2 g, 0.64
mmol), N-hydroxybenzotriazole (0.11 g, 0.70 mmol), and aniline
(0.06 g, 0.65 mmol) in DMF (10 ml) was added
1-ethyl-3-(3'-dimethylaminopropyl)carbodiimideHCl (0.148 g, 0.77
mmol) at 0.degree. C. After stirring for 16 hrs. the DMF was
removed, and ethyl acetate was added. This solution was washed with
sat. NaHCO.sub.3, brine and dried over sodium sulfate. The crude
residue was purified on silica gel using a gradient of 0% to 30%
ethyl acetate in dichloromethane as eluant to obtain 0.18 g (80%)
of 3-(2-isoquinolin-4-yl-ethyl)-N-phenyl-be- nzamide, S-1, as a
solid: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.16 (s, 1H), 8.30
(s, 1H), 8.04-7.99 (m, 2H), 7.76 (dd, 1H, J=8.34 Hz, J=8.37 Hz),
7.71-7.69 (m, 1H), 7.67-7.60 (m, 5H), 7.44-7.35 (m, 4H), 7.19-7.12
(m, 1H), 3.39-3.34 (m, 2H), 3.16-3.11 (m, 2H). MS (ESI) m/z 353
[M+H].sup.+. Anal. calc'd for C.sub.24H.sub.20N.sub.2O.0.2H.sub.2O:
C, 80.96; H, 5.78; N, 7.87. Found: C, 80.88; H, 5.85; N, 8.03.
EXAMPLE S-2
3-(2-Isoquinolin-4-yl-ethyl)-N-(3,3,5-trimethyl-cyclohexyl)-benzamide
[0447] 214
[0448] Example S-2 was prepared in a similar manner to that
described for S-1, except that
(.+-.)-cis/trans-3,3,5-trimethylcyclohexylamine was used in place
of aniline in step (f): .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
9.14 (s, 1H), 8.31 (s, 1H), 8.01 (m, 2H), 7.75 (dd, 1H, J=8.4 Hz,
J=8.4 Hz), 7.63 (dd, 1H, J=8.0 Hz, J=8.0 Hz), 7.56 (m, 1H), 7.52
(m, 1H), 7.36-7.28 (m, 2H), 5.72 (d, 1H, J=7.9 Hz), 4.16 (m, 1H),
3.37 (m, 2H), 3.10 (m, 2H), 2.90 (m, 1H), 1.78 (m, 2H), 1.38 (m,
1H), 1.03 (s, 3H), 0.97 (s, 3H), 0.92 (d, 3H, J=6.5 Hz), 0.84-0.64
(m, 3H). MS (ESI) m/z 401 [M+H].sup.+. Anal. calc'd for
C.sub.27H.sub.32N.sub.2O.0.1H.sub.2O: C, 80.60; H, 8.07; N, 6.96.
Found: C, 80.40; H, 8.20; N, 6.85.
EXAMPLE S-3
N-(4-Isopropyl-3-methyl-phenyl)-3-(2-isoquinolin-4-yl-ethyl)-benzamide
[0449] 215
[0450] Example S-3 was prepared in a similar manner to that
described for S-1, except that 4-isopropyl-3-methylaniline was used
in place of aniline in step (f): .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 9.16 (s, 1H), 8.31 (s, 1H), 8.04-7.99 (m, 2H), 7.76 (dd,
1H, J=8.3 Hz, J=8.4 Hz), 7.68-7.61 (m, 4H), 7.45-7.34 (m, 4H), 7.23
(m, 1H), 3.39-3.34 (m, 2H), 3.15-3.10 (m, 3H), 2.36 (s, 3H), 1.23
(d, 6H, J=6.9 Hz). MS (ESI) m/z 409 [M+H].sup.+. Anal. calc'd for
C.sub.28H.sub.28N.sub.2O.0.2H.sub.2O: C, 81.60; H, 6.95; N, 6.80.
Found: C, 81.51; H, 6.99; N, 6.85.
EXAMPLE S-4
3-(2-Isoquinolin-4-yl-ethyl)-N-(2-methyl-quinolin-6-yl)-benzamide
[0451] 216
[0452] Example S-4 was prepared in a similar manner to that
described for S-1, except that 6-amino-2-methylquinoline was used
in place of aniline in step (f): .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 9.16 (s, 1H), 8.44 (d, 1H, J=2.4 Hz), 8.29 (s, 1H),
8.07-7.97 (m, 5H), 7.79-7.71 (m, 2H), 7.68-7.61 (m, 3H), 7.46-7.38
(m, 2H), 7.29 (d, 1H, J=8.4 Hz), 3.40-3.35 (m, 2H), 3.17-3.10 (m,
2H), 2.74 (s, 3H). MS (ESI) m/z 418 [M+H].sup.+. Anal. calc'd for
C.sub.28H.sub.23N.sub.3O.0.4H.sub.2O: C, 79.18; H, 5.65; N, 9.89.
Found: C, 79.01; H, 5.86; N, 9.67.
EXAMPLE S-5
N-(3,5-Dibromo-4-methyl-phenyl)-3-(2-isoquinolin-4-yl-ethyl)-benzamide
[0453] 217
[0454] Example S-5 was prepared in a similar manner to that
described for S-1, except that 3,5-dibromo-4-methylaniline was used
in place of aniline in step (f): .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 9.16 (s, 1H), 8.25 (s, 1H), 8.01 (m, 2H), 7.88 (s, 2H),
7.76 (dd, 1H, J=8.16 Hz, J=8.51 Hz), 7.68-7.61 (m, 3H), 7.52 (m,
1H), 7.43-7.35 (m, 2H), 3.41-3.32 (m, 2H), 3.15-3.09 (m, 2H), 2.54
(s, 3H). MS (ESI) m/z 525 [M+H].sup.+. Anal. calcd for
C.sub.25H.sub.20N.sub.2OBr.sub.2* C.sub.2F.sub.3 OOH: C, 50.80; H,
3.32; N, 4.39. Found: C, 50.84; H, 3.40; N, 4.51.
EXAMPLE S-6
N-(4,6-Dimethyl-pyridin-2yl)-3(2
isoquinolin-4-yl-ethyl)-benzamide
[0455] 218
[0456] Example S-6 was prepared in a similar manner to that
described for S-1, except that 2-amino-4,6-dimethylpyridine was
used in place of aniline in step (f): .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 9.16 (s, 1H), 8.40 (m, 1H), 8.35 (s, 1H), 8.03
(m, 3H), 7.80-7.73 (m, 3H), 7.63 (dd, 1H, J=8.04 Hz, J=7.9 Hz),
7.44-7.36 (m, 2H), 6.79 (s, 1H), 3.39-3.34 (m, 2H), 3.16-3.10 (m,
2H), 2.45 (s, 3H), 2.37 (s, 3H). MS (ESI) m/z 382 [M+H].sup.+.
Anal. calcd for C.sub.25H.sub.23N.sub.3O.2C.sub.2F.sub.3
OOH.0.5H.sub.2O: C, 56.31; H, 4.24; N, 6.79. Found: C, 56.16; H,
4.17; N, 6.75.
EXAMPLE S-7
2-Chloro-4-fluoro-N.(4-isopropyl-3-methyl-phenyl)-5-(2-isoquinolin-4-yl-et-
hyl)-benzamide
[0457] 219
[0458] Example S-7 was prepared in a similar manner to that
described for S-1, except that ethyl
2-chloro-4-fluoro-5-bromobenzoate was used in place of ethyl
3-iodobenzoate in step (c), and 4-isopropyl-3-methylanilin- e was
used in place of aniline in step (f): .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 9.15 (s, 1H), 8.27 (s, 1H), 8.03(m, 2H),
7.81-7.76 (m, 2H), 7.64 (dd, 1H, J=7.17 Hz, J=7.11 Hz), 7.33 (d,
1H, J=7.91 Hz), 7.44-7.41 (m, 2H), 7.21-7.16 (m, 2H), 3.34-3.32 (m,
2H), 3.19-3.05 (m, 3H), 2.36 (s, 3H), 1.23(d, 6H, J=6.85 Hz). MS
(ESI) m/z 461 [M].sup.+. Anal. calcd for
C.sub.28H.sub.26ClFN.sub.2O: C, 72.95; H, 5.69; N, 6.08. Found: C,
72.70; H, 5.76; N, 6.03.
EXAMPLE S-8
2,4-Difluoro-N-(4-isopropyl-3-methyl-phenyl)-5-(2-isoquinolin-4-yl-ethyl)--
benzamide
[0459] 220
[0460] Example S-8 was prepared in a similar manner to that
described for S-1, except that methyl 2,4-difluoro-5-bromobenzoate
was used in place of ethyl 3-iodobenzoate in step (c), and
4-isopropyl-3-methylaniline was used in place of aniline in step
(f): .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.15 (s, 1H), 8.32
(s, 1H), 8.25 (m, 1H), 8.10-7.99 (m, 3H), 7.78 (dd, 1H, J=7.3 Hz,
J=8.4 Hz), 7.64 (dd, 1H, J=8.4 Hz, J=8.0 Hz), 7.47-7.40 (m, 2H),
7.24 (m, 1H), 6.92 (dd, 1H, J=11.8 Hz, J=11.8 Hz), 3.35-3.30 (m,
2H), 3.17-3.07 (m, 3H), 2.36 (s, 3H), 1.24 (d, 6H, J=6.9 Hz). MS
(ESI) m/z 445 [M].sup.+. Anal. calc'd for C.sub.28H.sub.26F.sub.3-
N.sub.2O: C, 75.66; H, 5.90; N, 6.30. Found: C, 75.42; H, 5.92; N,
6.22.
EXAMPLE T-1
2-Fluoro-N-(4-isopropyl-3-methyl-phenyl)-5-(2-isoquinolin-4-yl-ethyl)-benz-
amide
[0461] 221
[0462] (a)
5-Bromo-2-fluoro-N-(4-isopropyl-3-methyl-phenyl)-benzamide, T-1a,
was prepared from 4-isopropyl-3-methyl aniline and
4-bromo-2-fluorobenzoic acid in a manner similar to that described
for 3-(2-isoquinolin-4-yl-ethyl)-N-phenyl-benzamide in Example S-1,
step (f), except that
benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium
hexafluorophosphate was used in place of
1-ethyl-3-(3'-dimethylaminopropy- l)carbodiimideHCl and
N-hydroxybenzotriazole: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.29 (dd, J=10.5, 7.9 Hz, 1H), 8.24 (br s, 1H), 7.60 (m, 1H), 7.42
(m, 2H), 7.24 (d, J=8.4 Hz, 1H), 7.08 (dd, J=11.4, 8.7 Hz, 1H),
3.12 (septet, J=6.8 Hz, 1H), 2.36 (s, 3H), 1.27 (d, J=6.8 Hz,
6H).
[0463] (b)
2-Fluoro-N-(4-isopropyl-3-methyl-phenyl)-5-isoquinolin-4-ylethy-
nyl-benzamide, T-1b, was prepared in the manner similar to that
described in Example S-1, step (c) for ethyl
3-isoquinolin-4-ylethynyl-benzoate, S-1c, except that
5-bromo-2-fluoro-N-(4-isopropyl-3-methyl-phenyl)benzami- de, T-1a,
was used in place of ethyl 3-iodobenzoate. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 9.23 (s, 1H), 9.08 (s, 1H), 8.47 (dd, J=7.5,
2.2 Hz, 1H), 8.34 (m, 2H), 8.02 (d, J=8.1 Hz, 1H), 7.84 (m, 1H),
7.78 (m, 1H), 7.69 (m, 1H), 7.48 (m, 2H), 7.25 (m, 2H), 3.14
(septet, J=6.8 Hz, 1H), 2.38 (s, 3H), 1.24 (d, J=6.8 Hz, 6H).
[0464] (c)
2-Fluoro-N-(4-isopropyl-3-methyl-phenyl)-5-(2-isoquinolin-4-yl--
ethyl)-benzamide, T-1, was prepared in a manner similar to that
described in Example S-- l, step (d) for
3-(2-isoquinolin-4-yl-ethyl)-benzoic acid ethyl ester, S-1d:
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.14 (s, 1H), 8.37 (m,
1H), 8.29 (s, 1H) 8.05-7.99 (m, 3H), 7.77 (dd, 1H, J=8.3 Hz), 7.63
(dd, 1H, J=8.1 Hz), 7.49-7.45 (m, 2H), 7.23 (m, 2H), 7.07 (dd, 1H,
J=12 Hz), 3.37-3.32 (m, 2H), 3.18-3.07 (m, 3H), 2.37 (s, 3H), 1.23
(d, 6H, J=6.9 Hz). MS (ESI) m/z 427 [M+H].sup.+. Anal. calc'd for
C.sub.28H.sub.27FN.sub.2O: C, 78.85; H, 6.38; N, 6.57. Found: C,
78.91; H, 6.35; N, 6.39.
EXAMPLE U-1
N-(2-Methyl-quinolin-6yl)-3-(2-pyridin-3-yl-ethyl)Benzamide
Hydrochloride
[0465] 222
[0466] (a) A solution of 2-methyl-quinolin-6-ylamine (Maybridge,
836 mg, 5.29 mmol, 1.0 eq) and triethylamine (0.74 mL, 5.29 mmol,
1.0 eq) in dichloromethane (100 mL) was cooled to 0.degree. C. and
treated with 3-chloromethyl-benzoyl chloride (Aldrich, 0.75 mL,
5.29 mmol, 1.0 eq). A tan slurry forms within 30 minutes. After 1.0
hour, TLC (4% methanol/chloroform) gave no
2-methyl-quinolin-6-ylamine (R.sub.f 0.4), and a new spot with
R.sub.f 0.6. A 5% sodium bicarbonate solution (100 mL) was added to
the reaction mixture, and the aqueous layer was extracted with 10%
isopropanol/chloroform (3.times.100 mL) to give a yellow solid
(1.31 g). The product was washed with diethyl ether to give
3-chloromethyl-N-(2-methyl-quinolin-6-yl)-benzamide, U-1a, as a
yellow solid (1.28 g, 78%): HPLC R.sub.t=12.8 min.; TLC R.sub.f=0.6
(4% methanol/chloroform); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.56 (s, 1H), 8.46 (d, 1H, J=2.2 Hz), 8.22 (d, 1H, J=8.4
Hz), 8.07-7.90 (m, 4H), 7.70-7.69 (m, 1H), 7.58 (t, 1H, J=7.6 Hz),
7.40 (d, 1H, J=8.4 Hz), 4.88 (s, 2H), 2.65 (s, 3H); .sup.13C NMR
(75 MHz, DMSO-d.sub.6) .quadrature. 165.8, 158.0, 144.2, 138.4,
136.6, 136.5, 135.4, 132.5, 129.3, 128.4, 127.9, 126.7, 124.6,
123.0, 117.0, 116.8, 46.0, 24.7; MS (ESI) m/z 311 [M+H].sup.+.
[0467] (b) A solution of
3-chloromethyl-N-(2-methyl-quinolin-6-yl)-benzami- de, U-1a, (500
mg, 1.6 mmol, 1.0 eq) in acetone (250 mL) was heated to 55.degree.
C. and treated with sodium iodide (3.2 g, 150 mmol, 13 eq). After
2.0 h, no starting material was detected by HPLC, and the resulting
yellow solution was concentrated under reduced pressure. The
resulting residue was treated with water (100 mL) and the aqueous
layer was extracted with chloroform (3.times.100 mL). The combined
organic extracts were washed with Na.sub.2S.sub.2O.sub.3 (100 mL),
brine (100 mL), dried over magnesium sulfate, filtered and
concentrated under reduced pressure to give
3-iodomethyl-N-(2-methyl-quinolin-6-yl)-benzamide, U-1b, as a
yellow solid (615 mg, 96%): HPLC R.sub.t=13.6 min.; TLC R.sub.f=0.5
(5% methanol/chloroform); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.52 (s, 1H), 8.44 (d, 1H, J=2.1 Hz), 8.22 (d, 1H, J=8.5
Hz), 8.12-7.88 (m, 4H), 7.68-7.66 (m, 1H), 7.51 (t, 1H, J=7.7 Hz),
7.40 (d, 1H, J=8.5 Hz), 4.72 (s, 2H), 2.65 (s, 3H); MS (ESI) m/z
403 [M+H].sup.+.
[0468] (c) A solution of
3-iodomethyl-N-(2-methyl-quinolin-6-yl)-benzamide- , U-1b, (565 mg,
1.4 mg, 1.0 eq) in acetone (140 mL) was treated with
triphenylphosphine (1.80 g, 7.0 mmol, 5.0 eq) and heated to
55.degree. C. to give a yellow solution. After 18 hours, the
resulting slurry gave no starting material (R.sub.f 0.5) by TLC (5%
methanol/chloroform), only product with R.sub.f 0.0-0.3. The
solvent was removed under reduced pressure and the resulting solid
was washed with methyl-tert-butyl ether (4.times.25 mL) to give
N-(2-methyl-quinolin-6-yl)-3-[(triphenylphosphany-
l)-methyl]-benzamide iodide, U-1c, as a yellow solid (809 mg, 87%):
HPLC R.sub.t=14.4 min.; TLC R.sub.f=0.0-0.3 (5%
methanol/chloroform); .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta.
10.24 (s, 1H), 8.18 (d, 1H, J=1.1 Hz), 8.02 (d, 1H, J=8.5 Hz), 7.80
(d, 1H, J=7.8 Hz), 7.75-7.71 (m, 5H), 7.60-7.45 (m, 13H), 7.27-7.21
(m, 2H), 7.00 (d, 1H, J=7.7 Hz), 5.10 (d, 2H, J=15.7 Hz), 2.46 (s,
3H); MS (ESI) m/z 537 [M+H).sup.+.
[0469] (d) A solution of
N-(2-methyl-quinolin-6-yl)-3-[(triphenylphosphany-
l)-methyl]-benzamide iodide, U-1c, (600 mg, 0.90 mmol, 1.0 eq) in
THF (20 mL) was cooled to -78.degree. C. and treated with a 1.0 M
solution of sodium bis(trimethylsilyl)amide in THF (1.9 mL, 1.9
mmol, 2.1 eq) to give a dark orange solution. The solution was aged
for 30 min at -78.degree. C., then treated with
pyridine-3-carbaldehyde (93 .mu.L, 0.99 mmol, 1.1 eq). The orange
mixture was allowed to gradually warm to -30.degree. C. over 2 h,
removed from the cold bath and stirred at room temperature for 1.0
hour. The reaction was quenched with water (100 mL) and the aqueous
layer was extracted with chloroform (3.times.100 mL). The combined
organic extracts were washed with brine (100 mL), dried over
magnesium sulfate, filtered and concentrated under reduced pressure
to give a black oil (700 mg). The crude product was purified by
radial chromatography over silica gel using 3-4%
methanol/chloroform to give
trans-N-(2-methyl-quinolin-6-yl)-3-(2-pyridin-3-yl-vinyl)-benzamide,
U-1d, as a white solid (59 mg, 18%): HPLC R.sub.t=12.3 min.; TLC
R.sub.f=0.4 (5% methanol/chloroform); .sup.1H NMR (500 MHz,
DMSO-d.sub.6) .delta. 10.50 (s, 1H), 8.45-8.41 (m, 3H), 8.20 (d,
1H, J=8.4 Hz), 7.96 (dd, 1H, J=2.3, 9.1 Hz), 7.91-7.88 (m, 3H),
7.63-7.60 (m, 1H), 7.47 (t, 1H, J=7.5 Hz), 7.41-7.39 (m, 2H), 7.31
(dd, 1H, J=4.8, 8.0 Hz), 6.92 (d, 1H, J=12.3 Hz), 6.78 (d, 1H,
J=12.3 Hz), 2.64 (s, 3H); MS (ESI) m/z 366 [M+H].sup.+. Anal.
calc'd for C.sub.24H.sub.19N.sub.3O.0.2H.sub.2O: C, 78.11; H, 5.30;
N, 11.39. Found: C, 78.08; H, 5.38; N, 11.25.
[0470] (d) A solution of
trans-N-(2-methyl-quinolin-6-yl)-3-(2-pyridin-3-y-
l-vinyl)-benzamide, U-1d, (44 mg, 0.12 mmol) in methanol (3 mL) was
shaken vigorously under hydrogen (40 psi) with 5% palladium on
carbon for 24 h. The catalyst was filtered off, and the filtrate
was concentrated under reduced pressure to give a clear oil (50
mg). The crude product was purified by radial chromatography over
silica gel using 2-4% methanol/dichloromethane to give a clear oil.
The oil was dissolved in ethanol and treated with 0.1 mL
concentrated hydrochloric acid. The solvent was removed under
reduced pressure to give
N-(2-methyl-quinolin-6-yl)-3-(2-pyridin-3-yl-ethyl)-benzamide
hydrochloride, U-1, as a white solid (35 mg, 66%): HPLC R.sub.t
12.4 min.; .sup.1H NMR (500 MHz, DMSO-d.sub.6 w/ D.sub.2O) .delta.
(s, 1H), 8.96 (d, 1H, J=8.7 Hz), 8.82-8.81 (m, 2H), 8.74 (d, 1H,
J=5.4 Hz), 8.46 (d, 1H, J=8.0 Hz), 8.38 (dd, 1H, J=1.8, 9.2 Hz),
8.23 (d, 1H, J=9.2 Hz), 7.98-7.89 (m, 4H), 7.53-7.52 (m, 2H), 3.21
(t, 2H, J=7.8 Hz), 3.11 (t, 2H, J=7.7 Hz), 2.93 (s, 3H); .sup.13C
NMR (75 MHz, DMSO-d.sub.6 w/ D.sub.2O) .delta. 166.9, 156.6, 146.1,
144.8, 141.8, 141.3, 141.1, 140.2, 139.1, 135.0, 144.4, 132.8,
129.1, 128.5, 128.2, 127.6, 127.0, 126.1, 124.3, 121.4, 117, 35.7,
33.5, 20.9; MS (ESI) m/z 368 [M+H].sup.+. Anal. calc'd for
C.sub.24H.sub.21N.sub.3O.2HCl 0.3H.sub.2O: C, 64.66; H, 5.34; N,
9.43; Cl, 15.91. Found: C, 64.63; H, 5.40; N, 9.10; Cl, 15.62.
EXAMPLE U-2
N-(4-Isopropyl-3-methyl-phenyl)-3-(2-pyridin-3-yl-ethyl)-benzamide
[0471] 223
[0472] Example U-2 was prepared in a similar manner to that
described for U-1, except that 3-methyl-4-isopropylaniline was used
in place of 6-amino-2-methylquinoline in step (a): mp
143-144.degree. C.; HPLC R.sub.t=15.7 min.; TLC R.sub.f=0.4 (2%
methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.04 (s, 1H), 8.45 (s, 1H), 8.40 (d, 1H, J=4.5 Hz),
7.82-7.76 (m, 2H), 7.67 (d, 1H, J=7.5 Hz), 7.55-7.41 (m, 4H),
7.32-7.28 (m, 1H), 7.20 (d, 1H, J=8.3 Hz), 3.12-3.03 (m, 1H), 2.98
(s, 4H), 2.29 (s, 3H), 1.18 (d, 6H, J=6.8 Hz); HRMS (FAB) calcd for
C.sub.24H.sub.26N.sub.2O [M+H]+ 359.2123, found 359.2117.
EXAMPLE V-1
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-ylsulfanylmethyl]phenyl}-(3-bromo-4--
methyl)Benzamide
[0473] 224
[0474] (a) To a solution of 3-amino benzyl alcohol (123 mg, 1
mmol), 3-bromo-4-methylbenzoic acid (215 mg, 1 mmol) and
O-(7-azabenzotriazol-1-- yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HATU, 380 mg, 1 mmol) in 5 mL DMF was added
0.14 mL of triethylamine (1 mmol) and the reaction mixture stirred
at 50.degree. C. for 1 h. The solvent was removed in vacuo and the
residue was purified by chromatography to obtain
3-bromo-N-[3-hydroxymethyl)phenyl]-4-methylbenzamide, V-1a, (310
mg, 96%): .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.03 (s,
1H), 7.97 (s, 1H), 7.67 (d, 1H, J=7.5 Hz), 7.52 (s, 1H), 7.43 (d,
1H, J=7.6 Hz), 7.28 (d, 1H, J=6.9 Hz), 7.06 (m, 1H), 6.81-6.84 (d,
1H, J=7.1 Hz), 5.01 (br s, 1H), 4.27 (d, 2H, J=4.5 Hz), 2.20 (s,
3H); APCIMS m/z 338 [M+H].sup.+.
[0475] (b) To 3-bromo-N-[3-hydroxymethyl)phenyl]-4-methylbenzamide,
V-1a, (310 mg, 0.96 mmol) was added 5 mL of thionyl chloride and
the reaction mixture stirred for 15 min. Thionyl chloride was
removed in vacuo and the crude dissolved in ethyl acetate and
filtered through a plug of silica gel. The filtrate and washings
were combined and the solvent removed to obtain
3-bromo-N-[3-chloromethyl)phenyl]-4-methylbenzamide, V-1b (217 mg,
66%): .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.45 (s, 1H),
8.30 (s, 1H), 7.97-8.01 (m, 2H), 7.83 (d, 1H, J=8.3 Hz), 7.62 (d,
1H, J=8.3 Hz), 7.42-7.49 (m, 1H), 7.27 (d, 1H, J=7.5 Hz), 4.87 (s,
2H), 2.52 (s, 3H).
[0476] (c) To a solution of 97 mg (0.64 mmol) of
4-mercapto-1H-pyrazolo-[3- ,4-d]pyrimidine in 2 mL of DMF was added
NaHCO.sub.3 (80 mg) and 217 mg (0.64 mmol) of
3-bromo-N-[3-chloromethyl)phenyl]-4-methylbenzamide, V-1b. The
reaction mixture stirred at 50.degree. C. for 2 h. The solvent was
removed and water was added to the residue. The resulting solid was
filtered, washed with water and dried. The desired
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-ylsulfanylmethyl]phenyl}-(3-bromo-4-
-methy)benzamide, V-1, was obtained by silica gel column
chromatography purification (85 mg, 30%): .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.14 (s, 1H), 10.31 (s, 1H), 8.82 (s, 1H),
8.33 (s, 1H), 8.20 (s, 1H), 7.9-7.93 (m, 2H), 7.71 (d, 1H, J=6.8
Hz), 7.54 (d, 1H, J=7.9 Hz), 7.34 (dd, 1H, J=7.50, 7.9 Hz), 7.25
(d, 1H, J=6.8 Hz), 4.73 (s, 2H), 2.54 (s, 3H): APCIMS m/z 454
[M+H].sup.+.
EXAMPLE V-2
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-ylsulfanylmethyl]phenyl}-3,5-bis(Tri-
fluoromethyl)Benzamide
[0477] 225
[0478] Example V-2 was prepared in a similar manner to that
described for V-1, except that 3,5-bis(trifluoromethyl)benzoic acid
was used in place of 3-bromo-4-methylbenzoic acid in step (a):
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 14.05 (s, 1H), 10.60
(s, 1H), 8.73 (s, 1H), 8.52 (s, 2H), 8.31 (s, 1H), 8.23 (s, 1H),
7.82 (s, 1H), 7.63 (d, 1H, J=7.5 Hz), 7.20-7.32 (m, 2H), 4.66 (s,
2H); APCIMS m/z 498 [M+H].sup.+.
EXAMPLE V-3
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-(4-hydroxy-
-3-methoxy)Benzamide
[0479] 226
[0480] Example V-3 was prepared in a similar manner to that
described for V-1, except that 4-hydroxy-3-methoxybenzoic acid was
used in place of 3-bromo-4-methylbenzoic acid in step (a):.sup.1H
NMR (300 MHz, DMSO-d.sub.6) .delta. 14.05 (s, 1H), 9.93 (s, 1H),
9.63 (s, 1H), 8.72 (s, 1H), 8.23 (s, 1H), 7.81 (s, 1H), 7.58 (m,
1H), 7.43 (s, 2H), 7.13-7.25 (m, 2H), 6.78-6.80 (m, 1H), 4.66 (s,
2H), 3.27 (s, 3H); APCIMS rt/z 408 [M+H].sup.+.
EXAMPLE V-4
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-(4-hydroxy-
-3-t-butyl)Benzamide
[0481] 227
[0482] Example V-4 was prepared in a similar manner to that
described for V-1, except that 3-t-butyl-4-hydroxybenzoic acid was
used in place of 3-bromo-4-methylbenzoic acid in step (a): .sup.1H
NMR (300 MHz, DMSO-d.sub.6) .delta. 13.96 (s, 1H), 9.91 (s, 1H),
9.83 (s, 1H), 8.63 (s, 1H), 8.14 (s, 1H), 7.71 (s, 1H), 7.57 (s,
1H), 7.47-7.53 (m, 2H), 7.11 (dd, 1H, J=7.9, 8.0 Hz), 7.01 (d, 1H,
J=7.2 Hz), 6.69 (d, 1H, J=8.3 Hz), 4.52 (s, 2H), 1.22 (s, 9H);
APCIMS m/z 434 [M+H].sup.+.
EXAMPLE V-5
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-4-t-butylb-
enzamide
[0483] 228
[0484] Example V-5 was prepared in a similar manner to that
described for V-1, except that 4-t-butylbenzoic acid was used in
place of 3-bromo-4-methylbenzoic acid in step (a): .sup.1H NMR (300
MHz, DMSO-d.sub.6) .delta. 14.05 (s, 1H), 10.02 (s, 1H), 8.73 (s,
1H), 8.23 (s, 1H), 7.85 (s, 1H), 7.79 (d, 2H, J=8.3 Hz), 7.61 (d,
1H, J=7.6 Hz), 7.46 (d, 2H, J=8.3 Hz), 7.23 (dd, 1H, J=7.50, 7.9
Hz), 7.13 (d, 1H, J=7.6 Hz), 4.68 (s, 2H), 1.25 (s, 9H); APCIMS m/z
418 [M+H].sup.+.
EXAMPLE V-6
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-(4-phenoxy-
)Benzamide
[0485] 229
[0486] (a) To 3-aminobenzyl alcohol (1.23 g, 10 mmol) was added 20
mL of thionyl chloride and the reaction was stirred at room
temperature when a yellow solid separated out within five min. The
reaction was monitored by TLC for completion and excess thionyl
chloride was removed in vacuo to obtain the hydrochloride salt of
3-aminobenzylchloride (V-6a). To
4-mercapto-1H-pyrazolo-[3,4-d]pyrimidine (1.5 g, 10 mmol) in 5 mL
of DMF was added 4.6 mL of diisopropylethylamine (25 mmol) followed
by the addition of the hydrochloride salt of 3-aminobenzylchloride
(V-6a) and the reaction mixture stirred at 50.degree. C. for 1 h.
The solvent was removed in vacuo and the product
3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)su- lfanylmethyl]aniline,
V-6b, crystallized (1.1 g, 42%): .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.13 (s, 1H), 8.79 (s, 1H), 8.31 (s, 1H),
7.18 (dd, 1H, J=8.0, 8.3 Hz), 6.80 (d, 1H, J=8.3 Hz), 4.64 (s, 2H);
APCIMS m/z 258 [M+H].sup.+.
[0487] (b) To a solution of 64.5 mg (0.25 mmol) of
3-[(1H-pyrazolo[3,4-d]-- pyrimidin-4-yl)sulfanylmethyl]aniline,
V-6b, and 53.5 mg (0.25 mmol) of 4-phenoxybenzoic acid in 2 mL DMF
was added O-(7-azabenzotriazol-1-yl)-N,- N,N',N'-tetramethyluronium
hexafluorophosphate (HATU, 95 mg, 0.25 mmol) and triethylamine
(0.03 mL, 0.25 mmol). The reaction was stirred at 50.degree. C. for
1 h. After a conventional aqueous work-up,
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-(4-phenox-
y)benzamide, V-6, was obtained by chromatography on silica gel:
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.14.10 (s, 1H), 10.19 (s,
1H), 8.79 (s, 1), 8.30 (s, 1H), 7.97 (d, 2H, J=8.7 Hz), 7.90 (s,
1H), 7.67 (d, 1H, J=8.3 Hz), 7.46 (dd, 2H, J=7.60, 8.3 Hz), 7.31
(d, H, J=7.6 Hz), 7.267 (d, 1H, J=8.3 Hz), 7.19-7.22 (m, 2H),
7.07-7.12 (m, 3H), 4.70 (s, 2H); APCIMS m/z 454 [M+H].sup.+.
EXAMPLE V-6c
[0488] 0.1 M solutions of different acids, an amine template, HATU,
and triethylamine were prepared in anhydrous DMF. To each tube in
an array of 8.times.11 culture tubes (10.times.75 mm) was added 105
.mu.L (0.0105) of a different acid. To this was added 100 .mu.L
(0.01 mmol) of the amine solution, 105 .mu.L (0.0105 mmol) of the
triethylamine solution followed by 105 .mu.L (0.0105 mmol) of the
o-(7-azabenzotriazol-1-yl)-N,N,N',N'-te- tra-methyluronium
hexafluorophosphate solution. The reactions were stirred in a
heating block at 50.degree. C. for 3 h. The reaction mixtures were
transferred to a 1 mL 96-well-plates using a liquid handler. The
solvents were removed using the SpeedVac.TM. apparatus and the
crude reaction mixtures were redissolved in DMSO to give a final
theoretical concentration of 10 mM.
EXAMPLE V-6d
[0489] Using the general procedure described in Example V-6c above,
the following compounds were made:
23023123223323423523623723823924024124224-
32442452462472482492502512522532542552562572582592602612622632642652662672-
68269270271272273274275276277278279280281282283284
EXAMPLE V-7
N-{3-[(1H-pyrazolo[3,4-d]pyrimidin-4-ylsulfanyl)methyl]phenyl}-N'-[3,5-bis-
-(trifluoromethyl)Phenyl]Urea
[0490] 285
[0491] To a solution of 64.5 mg (0.25 mmol) of
3-[(1H-pyrazolo[3,4-d]-pyri- midin-4-yl)sulfanyl-methyl]aniline,
V-6b, and 3,5-bis(trifluromethyl)pheny- l isocyanate (61.2 mg, 0.25
mmol) in 2 mL DMF was added 20 mg of NaHCO.sub.3 and the reaction
stirred at 50.degree. C. for 1 h. After conventional aqueous
work-up, purification by silica gel chromatography provided
N-{3-[(1H-pyrazolo[3,4-d]pyrimidin-4-ylsulfanyl)methyl]phenyl}-N-
-[3,5-bis-(trifluoromethyl)phenyl]urea, V-7: .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 14.09 (s, 1H), 9.34 (s, 1H), 9.01 (s, 1H),
8.78 (s, 1H), 8.29 (s, 1H), 8.12 (s, 2H), 7.63 (s, 2H), 7.37 (d,
1H, J=7.5 Hz), 7.25 (dd, 1H, J=7.6, 8.3 Hz), 7.12 (d, 1H, J=7.6
Hz), 4.68 (s, 2H); APCIMS m/z 513 [M+H].sup.+.
EXAMPLE V-7a
[0492] 0.1 M solutions of the amine template, HATU, and isocyanate
were prepared in anhydrous DMF. To each tube in an array of
8.times.11 culture tubes (10.times.75 mm) was added 100 .mu.L (0.01
mmol) of the amine solution. To this was added 100 .mu.L (0.01
mmol) of a different isocyanate solution followed by the addition
of 10 mg of sodium bicarbonate. The reactions were stirred at
50.degree. C. for 2 h. The reaction mixtures were transferred to a
1 mL 96-well plate using a liquid handler. The solvents were
removed using the SpeedVac.TM. apparatus and the crude reaction
mixtures were redissolved in DMSO to give a final theoretical
concentration of 10 mM.
EXAMPLE V-7b
[0493] Using the general procedure described in Example V-7a above,
the following compounds were made:
28628728828929029129229329429529629729829- 9300
EXAMPLE V-8
N-{3-[(1H-pyrazolo[3,4-d]pyrimidin-4-ylsulfanyl)methyl]phenyl}-N'-(pyridin-
-3-yl)Urea
[0494] 301
[0495] Example V-8 was prepared in a manner similar to that
described in Example V-7, except that 3,5-bis(trifluromethyl)phenyl
isocyanate was replaced by 3-pyridyl isocyanate: HPLC R.sub.t=7.12
min.; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 14.12 (s, 1H),
8.79-8.84 (m, 3H), 8.59 (s, 1H), 8.30 (s, 1H), 8.17 (s, 1H), 792
(d, 1H, J=8.3 Hz), 7.59 (s, 1H) 7.21-7.38 (m, 3H), 7.09 (d, 1H,
J=7.2 Hz), 4.67 (s, 2H); APCIMS m/z 377 [M+H].sup.+.
EXAMPLE V-9
N-{3-[(1H-pyrazolo[3,4d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-(3,5-di-t-b-
utyl)Benzamide
[0496] 302
[0497] Example V-9 was prepared in a manner similar to that
described in Example V-6, except that 3,5-di-(t-butyl)benzoic acid
was used in place of 4-phenoxybenzoic acid in step (b): HPLC
R.sub.t=5.16 min.; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
14.12 (s, 1H), 10.19 (s, 1H), 8.80 (s, 1H), 8.30 (s, 1H), 7.87 (s,
1H), 7.72 (s, 2H), 7.66 (d, 2H, J=7.8 Hz), 7.60 (s, 1H), 7.31 (dd,
1H, J=7.60, 7.9 Hz), 7.22 (d, 1H, J=7.5 Hz), 4.70 (s, 2H), 1.33 (s,
18H); APCIMS m/z 474 [M+H].sup.+.
EXAMPLE V-10
3-Bromo-4-hydroxy-N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]-
phenyl}-benzamide
[0498] 303
[0499] Example V-10 was prepared in a manner similar to that
described in Example V-6, except that 3-bromo-4-hydroxybenzoic acid
was used in place of 4-phenoxybenzoic acid in step (b): .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 10.07 (s, 1H), 8.79 (s, 1H), 8.30
(s, 1H), 8.14 (d, 1H, J=2.1 Hz), 7.88 (s, 1H), 7.80 (dd, 1H, J=2.1,
8.1 Hz), 7.66 (d, 1H, J=8.6 Hz), 7.29 (dd, 1H. J=7.7, 7.8 Hz), 7.18
(d, 1H, J=7.6 Hz), 7.01 (d, 1H, J=8.5 Hz), 4.69 (s, 2H); APCIMS m/z
456 [M+H].sup.+.
EXAMPLE V-11
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl)-quinoline--
6-carboxamide
[0500] 304
[0501] Example V-11 was prepared in a manner similar to that
described in Example V-6, except that quinoline-6-carboxylic acid
was used in place of 4-phenoxybenzoic acid in step (b): .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 14.12 (s, 1H), 10.53 (s, 1H), 9.00
(dd, 1H, J=1.5, 4.2 Hz), 8.80 (s, 1H), 8.61 (d, 1H, J=1.9 Hz), 8.52
(d, 1H, J=8.0 Hz), 8.30 (s, 1H), 8.23 (dd, 1H, J=1.9, 8.7 Hz), 8.12
(d, 1H, J=8.7 Hz), 7.96 (s, 1H), 7.73 (d, 1H, J=7.9 Hz), 7.64 (dd,
1H, J=4.5, 8.4 Hz), 7.34 (dd, 1H, J=7.9, 8.0 Hz), 7.24 (d, 1H,
J=7.5 Hz), 7.05-7.12 (m, 1H), 4.72 (s, 2H); APCIMS m/z 413
[M+H].sup.+.
EXAMPLE V-12
5-Fluoro-N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]-phenyl}--
indole-2-carboxamide
[0502] 305
[0503] Example V-12 was prepared in a manner similar to that
described in Example V-6, except that 5-fluoroindole-2-carboxylic
acid was used in place of 4-phenoxybenzoic acid in step (b):
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 14.12 (s, 1H), 11.83
(s, 1H), 10.27 (s, 1H), 8.80 (s, 1H), 8.30 (s, 1H), 7.91 (s, 1H),
7.73 (d, 1H, J=8.3 Hz), 7.47 (s, 1H), 7.44 (dd, 1H, J=4.1, 4.60
Hz), 7.40 (d, 1H, J=1.9 Hz), 7.32 (dd, 1H, J=7.5, 8.0 Hz), 7.21 (d,
1H, J=7.1 Hz), 7.05-7.12 (m, 1H), 4.71 (s, 2H); APCIMS m/z 419
[M+H].sup.+.
EXAMPLE V-13
N-{3-[(1H-pyrazolo[3,4-d]-pyrimidin-4-yl)sulfanylmethyl]phenyl}-indole-6-c-
arboxamide
[0504] 306
[0505] Example V-13 was prepared in a manner similar to that
described in Example V-6, except that indole-6-carboxylic acid was
used in place of 4-phenoxybenzoic acid in step (b): .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 14.12 (s, 1H), 11.45 (s, 1H), 10.18
(s, 1H), 8.80 (s, 1H), 8.31 (s, 1H), 8.04 (s, 1H), 7.96 (s, 1H),
7.70 (d, 1H, J=7.9 Hz), 7.62 (s, 2H), 7.54 (s, 1H), 7.31 (dd, 1H,
J=7.9, 8.0 Hz), 7.18 (d, 1H, J=7.5 Hz), 7.30-6.52 (s, 1H), 4.70 (s,
2H); APCIMS m/z 401 [M+H].sup.+.
EXAMPLE V-14
[0506] The following compounds were made using the general
procedure described above in Example V-6c, except for the use of
different acids and amines, which yielded the products indicated
below (wherein for convenience, and as understood in the art, not
all hydrogen atoms have been expressly indicated for each carbon
and/or nitrogen atom).
3073083093103113123133143153163173183193203213223233243253263273283293303-
31332
EXAMPLE V-15
(R/S)-2-(2-methylphenyl)-N-{3-[(1H-pyrazolo[3,4-d]pyrimidin-4-ylsulfanyl)m-
ethyl)methyl}phenyl}butanamide
[0507] 333
[0508] Example V15 was prepared in a manner similar to that
described in Example V-6, except that 3-(o-tolyl)-butyric acid was
used in place of 4-phenoxybenzoic acid in step (b): .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta. 14.08 (s, 1H), 9.95 (s, 1H), 8.76
(s, 1H), 8.26 (s, 1H), 7.71 (s, 1H), 7.50-7.48 (d, 1H, J=8.31 Hz),
7.40-7.37 (dd, 1H, J=2.08, 8.31 Hz), 7.25-7.20 (d, 1H, J=7.74, 7.93
Hz), 7.16-7.09 (m, 4H), 4.64 (s, 2H), 3.76-3.72 (dd, 1H, J=5.48,
5.67 Hz), 2.38 (s, 3H), 1.99-1.92 (m, 1H), 1.63-1.60 (m, 1H),
0.90-0.80 (t, 3H, J=7.18, 7.37 Hz); APCIMS m/z 418 [M+H].sup.+.
EXAMPLE W-1
3-t-Butyl-4-hydroxy-N-{3-[5-(6-methoxy-pyridin-3-ylamino)-2H-pyrazol-3-ylm-
ethylsulfanyl]-phenyl}-benzamide
[0509] 334
[0510] (a) A solution of 3-t-butyl-4-hydroxy-benzoic acid (1.0 g,
5.2 mmol, 1.0 eq) in acetic anhydride (5.0 mL) was treated with
concentrated sulfuric acid (0.03 mL, 0.5 mmol, 0.1 eq). The clear
reaction mixture was warmed to 90.degree. C. After 18 h, the
resultant black solution was concentrated under reduced pressure
and treated with 1.0 M hydrochloric acid (100 mL). The aqueous
layer was extracted with ethyl acetate (3.times.50 mL) and the
combined organic extracts were washed with water (50 mL), brine (50
mL), dried over magnesium sulfate, filtered, and concentrated under
reduced pressure to give a brown solid (1.2 g). The crude product
was purified by radial chromatography over silica gel using 10-20%
ethyl acetate/cyclohexane with 0.1% acetic acid to give
4-acetoxy-3-t-butyl-benzoic acid, W-1a, as a white solid (309 mg,
26%): mp 123-125.degree. C.; TLC Rf=0.6 (20% ethyl
acetate/cyclohexane with 0.1% acetic acid); IR (KBr) 3418 (br),
1789, 1766 cm-1; 1H NMR (300 MHz, DMSO-d.sub.6) .delta. 8.12 (d,
1H, J=2.1 Hz), 8.06 (dd, 1H, J=8.4, 2.1 Hz), 7.36 (d, 1H, J=8.4
Hz), 2.39 (s, 3H), 1.36 (S, 9H).
[0511] (b). A slurry of 3-mercaptoaniline (1.00 mL, 9.42 mmol),
2-chloro-N-methoxy-N-methyl-acetamide and cesium carbonate (6.12 g,
18.84 mmol) in 5 mL of acetone was stirred for 18 h. The reaction
mixture was partitioned between ethyl acetate (30 mL) and sat.
sodium carbonate (2.times.50 mL) and the organic layer concentrated
to dryness to give an amber oil. Purification by chromatography on
silica gel using hexane/ethyl acetate (1:1) afforded 1.57 g (56%)
of 2-(3-aminophenylsulfanyl)-N-methoxy-N-methyl-acetamide, W-1b, as
a clear oil. HPLC R.sub.t=5.85 min; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.26 (s, 1H), 7.08 (t, 1H, J=8.1 Hz), 6.84-6.82
(m, 2H), 6.54 (d, 1H, J=8.7 Hz), 3.82 (s, 2H), 3.75 (s, 3H), 3.22
(s, 3H).
[0512] (c) To a solution of 0.24 g (0.81 mmol) of
2-(3-amino-phenylsulfany- l)-N-methoxy-N-methyl-acetamide, W-1b, in
dichloromethane (3 mL) was added 0.18 g (0.89 mmol) of
3-t-butyl-4-acetoxy-benzoic acid, W-1b, and EDC (0.18 g, 0.97
mmol). After 18 h, the reaction was partitioned between ethyl
acetate (30 mL) and sat. sodium bicarbonate (2.times.20 mL) and
washed with 1N HCl (2.times.20 mL). The organic layer was dried
over sodium sulfate and concentrated to a give yellow oil, which
was purified by chromatography on silica gel using hexane/ethyl
acetate (1:1) as eluant to afford
4-acetoxy-3-t-butyl-N-{3-[(methoxy-methyl-carbamoyl)-met-
hylsulfanyl]-phenyl}benzamide, W-1c, as a colorless oil, 0.31 g
(86%). HPLC R.sub.t=13.65 min.; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 8.41 (br s, 1H), 7.94 (d, 1H, J=2.1 Hz), 7.75 (t, 1H, J=1.8
Hz), 7.63 (dd, 1H, J=8.4, 2.1 Hz), 7.57-7.53 (m, 1H), 7.28-7.15 (m,
2H), 7.03 (d, 1H, J=8.4 Hz), 5.30 (s, 1H), 3.83 (s, 2H), 3.72 (s,
3H), 3.18 (s, 3H), 2.36 (s, 3 h), 1.36 (s, 9H).
[0513] LCESI: Calculated for C.sub.23H.sub.28N.sub.2O.sub.5S
(M+H.sup.+): 445, Found: 445.
[0514] (d). To a solution of 0.30 g (0.68 mmol) of
4-acetoxy-3-t-Butyl-N-{-
3-[(methoxy-methyl-carbamoyl)-methylsulfanyl]-phenyl}benzamide,
W-1c, in 7 mL of methanol/acetone/water (1:5:1) was added was
potassium carbonate (0.55 g, 1.35 mmol). The reaction mixture was
stirred for 1 h at room temperature and then partitioned between 1N
HCl (2.times.20 mL) and ethyl acetate (30 mL). The organic dried
over sodium sulfate and concentrated to give a yellow oil.
Trituration with diethyl ether (2.times.5 mL) gave 0.22 g (81%) of
3-t-butyl-4-hydroxy-N-{3-[(methoxy-methyl-carbamoyl)-meth-
ylsulfanyl]-phenyl}benzamide, W-1d, as a white solid: HPLC Rt=13.12
min.; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.04 (d, 1H, J=2.1
Hz), 8.00 (br s, 1H), 7.85 (dd, 1H, J=8.4, 2.4 Hz), 7.74 (d, 1H,
7.8 Hz), 7.49 (t, 1H, J=7.8 Hz), 7.39 (d, 1H, J=7.5 Hz), 7.02 (d,
1H, J=8.4 Hz), 4.13 (s, 2H), 3.95 (s, 3H), 3.41 (s, 3H), 1.64 (s,
9H). LCESI: Calculated for C.sub.21H.sub.26N.sub.2O.sub.4S
(M+H.sup.+): 403, Found: 403.
[0515] (e). To a solution of 0.24 g (1.31 mmol) of
N-(6-methoxy-pyridin-3-- yl)-thioacetamide in anhydrous THF (5 mL)
at -78.degree. C. was added dropwise 1.32 mL (2.64 mmol) of LDA
(2.0 M in THF). The reaction mixture was stirred for 0.25 h at
-78.degree. C., warmed to 0.degree. C. for 1 h, and then recooled
to -78.degree. C. To the resulting solution was added dropwise over
a 5 min period a solution of 0.17 g (0.41 mmol) of
3-t-butyl-4-hydroxy-N-{3-[(methoxy-methyl-carbamoyl)-methylsulfanyl]-phen-
yl}benzamide, W-1d, in 5 mL of THF. After 1 h at 0.degree. C., the
reaction was quenched with methanol/acetic acid (0.5 mL:0.5 mL) and
then partitioned between 30 mL of ethyl acetate and sat. sodium
carbonate (2.times.20 mL). The organic layer was concentrated to
give a yellow oil, which was purified by chromatography on silica
gel (1:1 hexane/ethyl acetate) to afford 0.22 g (96%) of
3-t-Butyl-4-hydroxy-N-{3-[3-(6-methoxy-
-pyridin-3-ylthiocarbamoyl)-2-oxo-propylsulfanyl]-phenyl}benzamide,
W-1e, as a pale yellow foam: HPLC Rt=13.12 min.; .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.23-8.18 (m, 1H), 8.08-7.98 (m, 2H),
7.86-7.70 (m, 2H), 7.58-7.40 (m, 2H), 7.28-7.15 (m, 2H), 6.82-6.68
(m, 2H). 3.94 (s, 3H), 3.89 (s, 2H), 2.74 (s, 2H). 1.36 (s, 9H).
LCESI: Calculated for C.sub.27H.sub.29N.sub.3O.sub.4S.sub.2
(M+H.sup.+): 524, Found: 524.
[0516] (f) To a solution of 0.18 g (0.33 mm ol) of
3-t-butyl-4-hydroxy-N-{-
3-[3-(6-methoxy-pyridin-3-ylthiocarbamoyl)-2-oxo-propylsulfanyl]-phenyl}-b-
enzamide, W-1e, in ethanol (2 mL) was added hydrazine mono-hydrate
(0.25 mL, 0.50 mmol) and acetic acid 0.025 mL, 0.28 mL). After 2 h,
the reaction solution was concentrated and the residue was purified
by chromatography (4:1 hexane:ethyl acetate) to afford 0.11 g (62%)
of
3-t-Butyl-4-hydroxy-N-{3-[5-(6-methoxy-pyridin-3-ylamino)-2H-pyrazol-3-yl-
methylsulfanyl]-phenyl}-benzamide, W-1: HPLC Rt=13.65 min.; .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.88 (br s, 1H), 7.73 (d, 1H,
J=2.4 Hz), 7.71 (s, 1H), 7.53 (dd, 1H, J=8.4, 2.4 Hz), 7.44 (br s,
1H), 7.17 (t, 1H, J=7.8 Hz), 7.02 (d, 1H, J=7.8 Hz), 7.71 (d, 1H,
J=8.4 Hz), 7.55 (d, 1H J=8.7 Hz), 5.64 (s, 1H), 5.39 (s, 1H), 4.03
(s, 2H), 3.71 (s, 3H), 1.33 (s, 9H); LCESI: Calculated for
C.sub.27H.sub.29N.sub.5O.sub.3S (M+H.sup.+): 504, Found: 504. Anal.
calc'd for C.sub.27H.sub.29N.sub.5O.s- ub.3S0.3 CH.sub.2Cl.sub.2:
C, 61.97; H, 5.64; N, 13.24. Found C, 61.78; H, 5.67; N, 13.16.
EXAMPLE W-2
3-t-Butyl-4-hydroxy-N-[3-(pyridin-3-ylmethylsulfanyl)-phenyl]-benzamide
[0517] 335
[0518] Example W-2 was prepared in a similar manner to that
described for the preparation of the intermediate
3-t-butyl-4-hydroxy-N-{3-[(methoxy-me-
thyl-carbamoyl)-methylsulfanyl]-phenyl}benzamide, W-1d, in example
W-1, except that 3-picolyl chloride hydrochloride was used in place
of 2-chloro-N-methoxy-N-methyl-acetamide in step (b): mp
95-100.degree. C.; HPLC R.sub.t=14.0 min.; TLC R.sub.f=0.5 (5%
methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.15 (s, 1H), 10.04 (s, 1H), 8.60 (s, 1H), 8.48 (d, 1H,
J=3.6 Hz), 7.88 (s, 1H), 7.84-7.72 (m, 3H), 7.64 (d, 1H, J=8.1 Hz),
7.40-7.29 (m, 2H), 7.10 (d, 1H, J=8.0 Hz), 6.92 (d, 1H, J=8.4 Hz),
4.32 (s, 2H), 1.46 (s, 9H); MS (ESI) m/z 393 [M+H].sup.+. Anal.
calc'd for C.sub.23H.sub.24N.sub.2O.sub.2S.0.5 MTBE: C, 72.63; H,
7.16; N, 6.22; S, 7.12. Found: C, 70.40; H, 6.86; N, 6.44; S,
7.17.
EXAMPLE W-3
3-t-Butyl-4-hydroxy-N-[3-(isoquinolin-4-ylmethylsulfnyl)-phenyl]-benzamide
[0519] 336
[0520] Example W-3, isolated as the hydrochloride salt as described
in Example K-2, was prepared in a similar manner to that described
for the preparation of the intermediate
3-t-butyl-4-hydroxy-N-{3-[(methoxy-methyl-
-carbamoyl)-methylsulfanyl]-phenyl}benzamide, W-1d, in example W-1,
except that 4-chloromethylisoquinoline hydrochloride, K-1c, was
used in place of 2-chloro-N-methoxy-N-methyl-acetamide in step (b):
mp 205-210.degree. C.; HPLC R.sub.t=15.2 min.; TLC R.sub.f=0.4 (5%
methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.28 (s, 1H), 10.15 (s, 1H), 9.81 (s, 1H), 8.66-8.59 (m,
3H), 8.32 (t, 1H, J=7.8 Hz), 8.11 (t, 1H, J=7.6 Hz), 7.95 (s, 1H),
7.88 (d, 1H, J=2.0 Hz), 7.83 (dd, 1H, J=8.0, 2.0 Hz), 7.75 (d, 1H,
J=9.0 Hz), 7.41 (t, 1H, J=8.0 Hz), 7.22 (d, 1H, J=7.7 Hz), 7.02 (d,
1H, J=8.4 Hz), 4.95 (s, 2H), 1.52 (s, 9H); MS (ESI) m/z 443
[M+H].sup.+. Anal. calc'd for C.sub.27H.sub.26N.sub.2O.sub.2S.HCl
0.2H.sub.2O: C, 66.45; H, 5.78; N, 5.743; S, 6.54. Found: C, 66.20;
H, 6.23; N, 5.37; S, 6.13.
EXAMPLE X-1
N-[3-(5-Bromo-pyridin-3-ylmethoxy)-phenyl]-3-1-butyl-4-hydroxy-benzamide
[0521] 337
[0522] (a) To a solution of 156 mg (1.43 mmol) of 3-aminophenol in
DMF at 0.degree. C. was added 115 mg (2.86 mmol) of 60% sodium
hydride dispersion in mineral oil. After 45 min, a slurry of 380 mg
(1.57 mmol) of 5-bromo-3-chloromethylpyridine hydrochloride, X-1a,
which was prepared from 5-bromo-3-(hydroxymethyl)pyridine (Hamel,
P. et al., J. Med. Chem., 40, 2866-2875 (1997)) according to the
procedure described in Example K-1, step (c), in DMF was added. The
reaction was allowed to slowly warm to room temperature over 3 h
and then partitioned between MTBE and sat. aq. ammonium chloride.
The organic layer was washed with brine, dried over MgSO.sub.4, and
concentrated. The residue was purified by radial chromatography
with a gradient of 0 to 1% methanol in 40% ethyl
acetate/cyclohexane to give 318 mg of
3-(5-bromo-pyridin-3-ylmethoxy)anil- ine, X-1b, as a clear oil:
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 8.68 (d, 1H, J=2.3 Hz),
8.64 (d, 1H, J=1.7 Hz), 8.10 (t, 1H, J=2.0 Hz), 6.91 (t, 1H, J=8.0
Hz), 6.17-6.22 (m, 3H), 5.10 (br s, 2H), 5.06 (s, 2H); MS (ESI) m/z
279/281 [M+H].sup.+.
[0523] (b) To a solution of 280 mg (1.0 mmol)
3-(5-bromo-pyridin-3-ylmetho- xy)aniline, X-1b, in dichloromethane
(10 mL) was sequentially added 326 mg (1.0 mmol) of
3-t-butyl-4-(t-butyl-dimethylsilanyloxy)-benzoyl chloride, X-1c,
(Trova, M. P. et al., J. Med. Chem., 36, 580-590 (1993)) and
triethylamine (0.15 mL, 1.1 mmol). The resultant clear solution was
stirred for 18 h at room temperature and then the solvent was
removed under reduced pressure. The reaction mixture was treated
with 5% sodium bicarbonate (100 mL) and extracted with 10%
isopropanol/chloroform (3.times.50 mL). The combined organic
extracts were washed with brine (50 mL), dried over magnesium
sulfate, filtered, and concentrated under reduced pressure to give
an off-white solid (569 mg). The crude product was purified by
radial chromatography over silica gel using 1-2%
methanol/dichloromethane to give
N-[3-(5-bromo-pyridin-3-ylmethoxy)-pheny-
l]-3-t-butyl-4-(t-butyl-dimethyl-silanyloxy)-benzamide, X-1d, as a
white solid (456 mg, 80%): HPLC R.sub.t=20.6 min; TLC R.sub.f=0.8
(4% methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 9.91 (s, 1H), 8.53-8.51 (m, 2H), 8.00 (t, 1H, J=2.0 Hz),
7.64 (d, 1H, J=2.3 Hz), 7.56 (dd, 1H, J=8.5, 2.2 Hz), 7.41 (t, 1H,
J=2.0 Hz), 7.19-7.16 (m, 1H), 7.09 (t, 1H, J=8.1 Hz), 6.78 (d, 1H,
J=8.4 Hz), 6.60 (dd, 1H, J=8.0, 1.6 Hz), 5.00 (s, 2H), 1.23 (s,
9H), 0.86 (s, 9H), 0.20 (s, 6H); MS (ESI) m/z 569/571
[M+H].sup.+.
[0524] (c) To a solution of 100 mg (0.18 mmol)
N-[3-(5-bromo-pyridin-3-ylm-
ethoxy)-phenyl]-3-t-butyl-4-(t-butyl-dimethyl-silanyloxy)-benzamide,
X-1d, in THF (7 mL) at 0.degree. C. was added a 1.0 M solution of
tetrabutylammonium fluoride in THF (0.27 mL, 0.27 mmol). The
slightly yellow reaction mixture was warmed to room temperature
over several hours and stirred an additional 15 h. The cloudy
reaction mixture was concentrated under reduced pressure to give a
clear oil. The crude product was purified by radial chromatography
over silica gel using 2-4% methanol/dichloromethane to give
N-[3-(5-bromo-pyridin-3-ylmethoxy)-pheny-
l]-3-t-butyl-4-hydroxy-benzamide, X-1, as a white solid (75 mg,
91%): mp 105-111.degree. C.; HPLC R.sub.t=14.7 min.; TLC
R.sub.f=0.4 (4% methanol/dichloromethane); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.11 (s, 1H), 10.01 (s, 1H), 8.74-8.72 (m,
2H), 8.21 (t, 1H, J=2.0 Hz), 7.78 (d, 1H, J=2.1 Hz), 7.72 (dd, 1H,
J=8.3, 2.2 Hz), 7.61 (t, 1H, J=2.0 Hz), 7.40-7.37 (m, 1H), 7.29 (t,
1H, J=8.1 Hz), 6.91 (d, 1H, J=8.4 Hz), 6.79 (dd, 1H, J=7.8, 2.1
Hz), 5.20 (s, 2H), 1.43 (s, 9H); .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. 166.1, 159.5, 158.4, 150.1, 147.9, 141.2,
138.3, 135.4, 135.3, 129.7, 127.2, 127.1, 125.4, 120.5, 115.9,
113.5, 109.9, 107.2, 66.3, 34.8, 29.5; MS (ESI) m/z 455/457
[M+H].sup.+. Anal. calc'd for C.sub.23H.sub.23BrN.sub.2O.sub.3: C,
60.67; H, 5.09; Br, 17.55; N, 6.15. Found: C, 60.63; H, 5.24; Br,
17.69; N, 6.01.
EXAMPLE X-2
4-Acetoxy-3-t-butyl-N-[3-(pyridin-3-ylmethoxy)phenyl]-benzamide
[0525] 338
[0526] Example X-2 was prepared from
3-(pyridin-3-ylmethoxy)aniline, X-2a, prepared from
3-picolylchloride hydrochloride and 3-hydroxyaniline as described
in Example X-1, step (a), and 4-acetoxy-3-t-butylbenzoic acid,
W-1a, according to the procedure described in Example W-1, step
(c): mp 58-62.degree. C.; HPLC R.sub.t=14.4 min.; TLC R.sub.f=0.3
(4%. methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.25 (s, 1H), 8.70 (s, 1H), 8.56 (d, 1H, J=4.3 Hz),
7.91-7.80 (m, 3H), 7.58-7.56 (m, 1H), 7.46-7.21 (m, 4H), 6.81-6.78
(m, 1H), 5.16 (s, 2H), 2.37 (s, 3H), 1.36 (s, 9H); MS (ESI) m/z 419
[M+H].sup.+. Anal. calc'd for
C.sub.25H.sub.26N.sub.2O.sub.4.0.4H.sub.2O: C, 70.53; H, 6.35; N,
6.58. Found: C, 70.87; H, 6.28; N, 6.59.
EXAMPLE X-3
4-Acetoxy-3-t-butyl-N-[3-(isoquinolin-4-ylmethoxy)phenyl]-benzamide
[0527] 339
[0528] Example X-3 was prepared from
3-(isoquinolin-4-ylmethoxy)aniline, prepared from
4-chloromethylisoquinoline hydrochloride, K-1c, and
3-hydroxyaniline as described in Example X-1, step (a), and
4-acetoxy-3-t-butylbenzoic acid, W-1a, according to the procedure
described in Example W-1, step (c): mp 83-86.degree. C.; HPLC
R.sub.t=15.6 min.; TLC R.sub.f=0.3 (1% methanol/dichloromethane);
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.26 (s, 1H), 9.37 (s,
1H), 8.67 (s, 1H), 8.21 (d, 1H, J=7.9 Hz), 8.14 (d, 1H, J=8.8 Hz),
7.91-7.73 (m, 4H), 6.60 (t, 1H, J=2.2 Hz), 7.40-7.37 (m, 1H), 7.30
(t, 1H, J=8.1 Hz), 7.22 (d, 1H, J=8.3 Hz), 6.90 (dd, 1H, J=7.6, 2.2
Hz), 5.56 (s, 2H), 2.36 (s, 3H), 1.36 (s, 9H); MS (ESI) m/z 469
[M+H].sup.+. Anal. calc'd for
C.sub.29H.sub.28N.sub.2O.sub.4.0.2H.sub.2O: C, 73.77; H, 6.06; N,
5.93. Found: C, 73.46; H, 6.38; N, 5.82.
EXAMPLE X-4
3-t-Butyl-4-hydroxy-N-[3-(pyridin-3-ylmethoxy)-phenyl]-benzamide
[0529] 340
[0530] Example X-4 was prepared from
4-acetoxy-3-t-butyl-N-[3-(pyridin-3-y- lmethoxy)phenyl]-benzamide,
X-2, in a manner similar to that described in Example W-1, step
(d): mp 104-107.degree. C.; HPLC R.sub.t=13.6 min; TLC R.sub.f=0.5
(5% methanol/dichloromethane); .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta. 10.07 (s, 1H), 9.96 (s, 1H), 8.69 (s, 1H), 8.56 (d, 1H,
J=.4.0 Hz), 7.90 (d, 1H, J=7.9 Hz), 7.74-7.67 (m, 2H), 7.57 (s,
1H), 7.47-7.42 (m, 1H), 7.35-7.33 (m, 1H), 7.24 (t, 1H, J=8.0 Hz),
6.87 (d, 1H, J=8.3 Hz), 6.77-6.74 (m, 1H), 5.15 (s, 2H), 1.40 (s,
9H); .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. 166.1, 159.5,
158.6, 149.5, 149.4, 141.2, 136.0, 135.4, 133.0, 130.0, 127.2,
127.1, 125.4, 124.0, 115.9, 113.3, 109.9, 107.2, 67.2, 34.5, 29.5;
MS (ESI) m/z 375 [M-H].sup.-.
[0531] Anal. calc'd for C.sub.23H.sub.24N.sub.2O.sub.3.0.5 MTBE: C,
72.83; H, 7.19; N, 6.66. Found: C, 72.61; H, 7.12; N, 6.66.
EXAMPLE X-5
3-t-Butyl-4-hydroxy-N-[3-(isoquinolin-4-ylmethoxy)-phenyl]-benzamide
[0532] 341
[0533] Example X-5 was prepared from
4-acetoxy-3-t-butyl-N-[3-(isoquinolin-
-4-ylmethoxy)phenyl]-benzamide, X-3, in a manner similar to that
described in Example W-1, step (d): HPLC R.sub.t=14.8 min; TLC
R.sub.f=0.4 (4% methanol/dichloromethane); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 10.09 (s, 1H), 9.97 (s, 1H), 9.34 (s, 1H),
8.65 (s, 1H), 8.20 (d, 1H, J=8.2 Hz), 8.13 (d, 1H, J=7.9 Hz),
7.90-8.86 (m, 1H), 7.78-7.59 (m, 4H), 7.38-7.35 (m, 1H), 7.26 (t,
1H, J=8.0 Hz), 6.85 (d, 2H, J=8.6 Hz), 5.54 (s, 2H), 1.38 (s, 9H);
HRMS (FAB) calcd for C.sub.27H.sub.26N.sub.2O.sub.- 3 [M+H]+
427.2022, found 427.2020.
EXAMPLE Y-1
1-[3-(pyridin-3-ylmethoxy)phenylcarbamoyl]pyrrolidine
[0534] 342
[0535] A solution of 100 mg (0.50 mmol) of
3-(pyridin-3-ylmethoxy)aniline, X-2a, and 0.076 mL (0.55 mmol) of
triethylamine in 2 mL of dichloromethane was added dropwise to a
solution of triphosgene (54 mg, 0.18 mmol) in 2 mL of
dichloromethane. After 20 min, a solution of pyrrolidine (0.042 mL)
and triethylamine (0.076 mL) in 2 mL of dichloromethane was added
to the reaction mixture. After 2 h, the reaction was partitioned
between dichloromethane and 5% aq. sodium bicarbonate. The organic
layer was washed with water and brine, dried over MgSO.sub.4 and
concentrated. The residue was purified by radial chromatography
with a gradient of 0 to 2% methanol in 1:1 ethyl
acetate:cyclohexane to give 81 mg (55%) of
1-[3-(pyridin-3-ylmethoxy)phen- ylcarbamoyl]pyrrolidine, Y-1, as a
white solid: mp 138-140.degree. C.; HPLC R.sub.t=9.7 min.; TLC
R.sub.f=0.3 (5% methanol/dichloromethane); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 8.66 (d, 1H, J=1.7 Hz), 8.54 (dd, 1H, J=4.7,
1.5 Hz), 8.08 (s, 1H), 7.88-7.85 (m, 1H), 7.45-7.41 (m, 1H),
7.35-7.34 (m, 1H), 7.16-7.09 (m, 2H), 6.61-6.58 (m, 1H), 5.09 (s,
2H), 3.40-3.31 (m, 4H), 1.87-1.83 (m, 4H); .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. 158.5, 154.1, 149.4, 149.3, 142.3, 135.9,
133.1, 129.4, 123.9, 112.4, 108.0, 106.3, 67.1, 46.0, 25.4; MS
(ESI) m/z 298 [M+H].sup.+. Anal. calc'd for
C.sub.17H.sub.19N.sub.3O.sub.2: C, 68.67; H, 6.44; N, 14.13. Found:
C, 68.41; H, 6.50; N, 13.89.
EXAMPLE Y-2
4-[3-(pyridin-3-ylmethoxy)phenylcarbamoyl]morpholine
[0536] 343
[0537] Example Y-2 was prepared in a manner similar to that
described in Example Y-1, except that morpholine was used in place
of pyrrolidine: mp 58-62.degree. C.; HPLC R.sub.t=8.7 min.; TLC
R.sub.f=0.3 (5% methanol/methylene chloride); .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 8.96 (s, 1H), 8.84 (d, 1H, J=5.3 Hz), 8.63
(s, 1H), 8.50 (d, 1H, J=7.8 Hz), 8.00-7.95 (m, 1H), 7.36 (s, 1H),
7.16 (t, 1H, J=8.1 Hz), 7.07 (d, 1H, J=7.9 Hz), 6.65-6.63 (m, 1H),
5.26 (s, 2H), 3.60 (t, 4H, J=4.5 Hz), 3.42 (t, 4H, J=4.5 Hz);
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. 157.7, 155.0, 143.8,
141.9, 141.1, 136.9, 129.2, 126.8, 112.6, 108.0, 106.2, 66.0, 65.4,
44.2; MS (ESI) m/z 314 [M+H].sup.+. Anal. calc'd for
C.sub.17H.sub.19N.sub.3O.sub.3 HCl-0.3H.sub.2O: C, 57.48; H, 5.85;
Cl, 9.98; N, 11.83. Found: C, 57.05; H, 5.83; Cl, 9.99; N,
11.50.
EXAMPLE Z-1
3-[{6-Methoxy-7-(2-methoxyethoxy)Cinnolin-4-yl}sulfanylmethyl]-N-phenyl-be-
nzamide
[0538] 344
[0539] (a) To a solution of 0.30 g (1.33 mmol) of
3-(chloromethyl)-N-pheny- lbenzamide, Z-1a, which prepared in a
manner similar to that described in Example A-1, step (a), in 6 mL
of ethanol was added 0.20 g (2.65 mmol) of thiourea, and the
mixture was heated to 80.degree. C. After 3 h, 1.0 mL (3 mmol) of
2N aq. sodium hydroxide was added, and heating at 80.degree. C. was
continued. After an additional 2.5 h, the reaction was cooled to
room temperature and 10 mL of water was added. The mixture was
extracted with ethyl acetate, and the aqueous layer was neutralized
with 1N aq. HCl and extracted again with ethyl acetate. The
combined organic layers were washed with water and with brine,
dried over sodium sulfate, and concentrated. The residue was
purified by chromatography on silica gel, eluting with a gradient
of 15% to 30% ethyl acetate in hexane, to provide 174 mg of
3-(mercaptomethyl)-N-phenylbenzamide, Z-1b: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.85 (s, 2H), 7.75 (m, 1H), 7.67 (m, 2H), 7.53
(m, 1H), 7.40, (m, 3H), 7.17 (m, 1H), 3.80 (d, 2H), 1.83 (t,
1H).
[0540] (b) To a solution of 50 mg (0.186 mmol) of
4-chloro-6-methoxy-7-(2-- methoxyethoxy)cinnoline, Z-1c, (PCT
application WO 97/34876, p.51) and 42.5 mg (0.189 mmol) of
3-(mercaptomethyl)-N-phenylbenzamide, Z-1b, in 1.2 mL of
isopropanol was added 12.3 mg of potassium hydroxide in 1.2 mL of
ethanol. The mixture was heated to 40.degree. C. for 45 min, then
cooled to room temperature. The precipitate was collected by
filtration and air-dried to give 44.7 mg (47%) of
3-[{6-methoxy-7-(2-methoxyethoxy)c-
innolin-4-yl}sulfanylmethyl]-N-phenyl-benzamide, Z-1: .sup.1H NMR
(300 MHz, DMSO-d.sub.6) 810.29 (s, 1H), 9.19 (s, 1H), 8.10 (s, 1H),
7.89 (m, 1H), 7.76 (m, 4H), 7.54 (m, 1H), 7.38 (m, 2H), 7.09 (m,
2H), 4.72 (s, 2H), 4.38 (m, 2H), 3.98 (s, 3H), 3.77 (m, 2H), 3.32
(s, 3H). MSESI.sup.(+): M+H.sub.+ 476, M+Na.sub.+ 498, M+K.sub.+
514.
EXAMPLE AA-1
3-[2-(6-Acetylamino-pyridin-3-yl)-ethyl]-N-(4-piperazin-1-yl-3-trifluorome-
thylphenyl)-benzamide dihydrochloride
[0541] 345
[0542] (a) To a solution of 5-bromo-pyridin-2-ylamine (Aldrich, 2.0
g, 11.6 mmol, 1.0 eq) in tetrahydrofuran (100 mL) was added acetic
anhydride (3.0 mL, 31.8 mmol, 2.7 eq) and triethylamine (1.8 mL,
12.8 mmol, 1.1 eq). After 3 days, the solvent was removed and the
crude reaction mixture was dissolved in ethyl acetate and
sequentially washed with aqueous 5% sodium bicarbonate and brine.
The crude product was dried over magnesium sulfate to give
N-(5-bromo-pyridin-2-yl)-acetamide, AA-1a, as a white solid (2.5 g,
100%): HPLC R.sub.t 8.4 min.; TLC R.sub.f 0.7 (2%
methanol-dichloromethane); .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 10.62 (s, 1H), 8.42 (dd, 1H, J=2.5, 0.7 Hz), 8.06 (d, 1H,
J=9.0 Hz), 7.98 (dd, 1H, J=9.0, 2.4 Hz), 2.09 (s, 3H); .sup.13C NMR
(DMSO-d.sub.6, 75 MHz) .delta. 169.8, 151.4, 148.8, 140.8, 115.3,
113.5, 24.2; MS m/z 215/217 (M+H).sup.+.
[0543] (b) To a solution of N-(5-bromo-pyridin-2-yl)-acetamide,
AA-1a, (2.2 g, 10.2 mmol, 1.0 eq) in degassed N,N-dimethylformamide
(100 mL) was added diisopropylethylamine (3.6 mL, 20.4 mmol, 2.0
eq), copper(I) iodide (155 mg, 0.82 mmol, 0.08 eq),
dichlorobis(triphenylphosphine)palladium(II- ) (286 mg, 0.41 mmol,
0.04 eq) and triethyl-ethynyl-silane (Aldrich, 3.7 mL, 20.4 mmol,
2.0 eq). The resultant red solution was warmed to 115.degree. C.
for 2 days. The crude reaction mixture was poured into water and
extracted with methyl-tert-butyl ether/ethyl acetate. The organic
layer was washed with brine, dried over magnesium sulfate and
purified over silica, which was eluted with 10% ethyl
acetate-cyclohexane, to give
N-(5-triethylsilanylethynyl-pyridin-2-yl)-ac- etamide, AA-1b, as a
beige solid (1.2 g, 44%): HPLC R.sub.t 17.8 min.; TLC R.sub.f 0.5
(20% ethyl acetate-cyclohexane); .sup.1H NMR (DMSO-d.sub.6, 300
MHz) .delta. 10.69 (s, 1H), 8.40 (d, 1H, J=1.7 Hz), 8.08 (d, 1H,
J=8.6 Hz), 7.84 (dd, 1H, J=8.7, 2.3 Hz), 2.10 (s, 3H), 1.01 (t, 6H,
J=7.8 Hz), 0.68 (q, 9H, J=7.8 Hz); MS (ESI) m/z 275
(M+H).sup.+.
[0544] (c) To a solution of
N-(5-triethylsilanylethynyl-pyridin-2-yl)-acet- amide, AA-1b, (1.0
g, 3.6 mmol, 1.0 eq) in tetrahydrofuran (40 mL) was added a
solution of tetrabutylammonium fluoride (4.0 mL, 1.0 M, 4.0 mmol,
1.1 eq). The resultant amber solution was stirred at room
temperature for 18 h. The solvent was removed under reduced
pressure and the crude reaction mixture (1.1 g, tan solid) was
dissolved in ethyl acetate and passed through a silica plug, which
was eluted with ethyl acetate. The resultant yellow solid (1.0 g)
was washed with 10% MTBE-cyclohexane to give
N-(5-ethynyl-pyridin-2-yl)-acetamide, AA-1c, as a tan solid (491
mg, 84%): HPLC R.sub.t 7.1 min.; TLC R.sub.f 0.5 (30% ethyl
acetate-cyclohexane); .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta.
10.65 (s, 1H), 8.41 (d, 1H, J=1.6 Hz), 8.08 (d, 1H, J=8.7 Hz), 7.86
(dd, 1H, J=8.6, 2.2 Hz), 4.29 (s, 1H), 2.10 (s, 3H); .sup.13C NMR
(DMSO-d.sub.6, 75 MHz) .delta. 169.5, 151.6, 150.8, 141.0, 113.6,
112.7, 82.6, 80.5, 23.9; MS (ESI) m/z 161 (M+H).sup.+.
[0545] (d) To a solution of 2-chloro-5-nitrobenzotrifluoride
(Lancaster, 16.5 mL, 112 mmol, 1.0 eq) and piperazine-1-carboxylic
acid tert-butyl ester (Aldrich, 2.5 g, 134 mmol, 1.2 eq) in
N,N-dimethylformamide (225 mL) was added potassium carbonate (46.3
g, 336 mmol, 3.0 eq). The resultant red solution was warmed to
90.degree. C. for 24 h. The crude product was poured into ice water
(1.7 L) and extracted with ethyl acetate (3.times.300 mL). The
combined organic extracts were diluted with methyl-tert-butyl ether
(1.0 L) and sequentially washed with water and brine. The organic
layer was dried over magnesium sulfate, filtered and concentrated
under reduced pressure to give 4-(nitro-trifluoromethyl-phen-
yl)-piperazine-1-carboxylic acid tert-butyl ester, AA-1d, as an
orange solid (43.6 g, 104%): HPLC R.sub.t 17.0 min.; TLC R.sub.f
0.5 (20% ethyl acetate-cyclohexane); .sup.1H NMR (DMSO-d.sub.6, 300
MHz) .delta. 8.46-8.40 (m, 2H), 7.62 (d, 1H, J=8.9 Hz), 3.48 (t,
4H, J=4.8 Hz), 3.06 (t, 4H, J=5.0 Hz), 1.43 (s, 9H); MS (ESI) m/z
276 (M+H-BOC).sup.+.
[0546] (e) To a solution of
4-(nitro-trifluoromethyl-phenyl)-piperazine-1-- carboxylic acid
tert-butyl ester, AA-1d, (43.6 g, 116 mmol) and 10% palladium on
carbon (4.3 g) in ethyl acetate (1.2 L) was added hydrogen (1 atm).
The resultant slurry was stirred for 18 h, filtered through celite
and concentrated under reduced pressure to give
4-[4-(t-butoxycarbonyl)piperazin-1yl]-3-trifluoromethylaniline,
AA-1e, as a yellow solid (40 g, 92%): HPLC R.sub.t 14.8 min.; TLC
R.sub.f 0.1 (10% ethyl acetate-cyclohexane); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 7.22 (d, 1H, J=8.5 Hz), 6.82 (d,
1H, J=2.6 Hz), 6.75 (dd, 1H, J=8.5, 2.5 Hz), 5.37 (s, 2H), 3.37
(br. s, 4H), 2.66 (t, 4H, J=4.8 Hz), 1.41 (s, 9H); MS (ESI) m/z 346
(M+H).sup.+.
[0547] (f) To a solution of
4-[4-(t-butoxycarbonyl)piperazin-1yl]-3-triflu- oromethylaniline,
AA-1e, (10.0 g, 29.0 mmol, 1.0 eq) and 3-iodobenzoic acid (Aldrich,
8.6 g, 34.8 mmol, 1.2 eq) in ethyl acetate (300 mL) and
dichloromethane (300 mL) was added
1-(3-dimethylaminopropyl)-3-ethylcarbo- diimide hydrochloride (6.7
g, 34.8 mmol, 1.2 eq). The resultant brown solution was stirred for
18 h. The solvent was removed under reduced pressure and the crude
product was purified over silica, which was eluted with 10-20%
ethyl acetate-cyclohexane, to give 4-({[1-(3-iodo-phenyl)-met-
hanoyl]-amino}-trifluoromethyl-phenyl)-piperazine-1-carboxylic acid
tert-butyl ester, AA-1f, as a beige solid (10.3 g, 62%): HPLC
R.sub.t 19.7 min.; TLC R.sub.f 0.5 (30% ethyl acetate-cyclohexane);
.sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.53 (s, 1H), 8.32 (s,
1H), 8.14 (d, 1H, J=2.3 Hz), 8.04 (dd, 1H, J=8.7, 2.3 Hz),
7.99-7.96 (m, 2H), 7.60 (d, 1H, J=8.8 Hz), 7.36 (t, 1H, J=7.8 Hz),
3.44 (br. s, 4H), 2.51 (t, 4H, J=4.6 Hz), 1.43 (s, 9H); MS (ESI)
m/z 576 (M+H).sup.+.
[0548] (g)
4-[({1-[3-(6-Acetylamino-pyridin-3-ylethynyl)-phenyl]-methanoyl-
}-amino)-trifluoromethyl-phenyl]-piperazine-1-carboxylic acid
tert-butyl ester, AA-1g, was prepared in the manner similar to that
described in example S-1, step (c) for ethyl
3-isoquinolin-4-ylethynyl-benzoate, S-1c, except that
4-({[1-(3-iodo-phenyl)-methanoyl]-amino}-trifluoromethyl-phen-
yl)-piperazine-1-carboxylic acid tert-butyl ester, AA-1f, was used
in place of 3-iodobenzoate and
N-(5-ethynyl-pyridin-2-yl)-acetamide, AA-1c, was used in place of
4-ethynyl-isoquinoline, S-1b: HPLC R.sub.t 18.1 min.; TLC R.sub.f
0.5 (2% methanol-dichloromethane); .sup.1H NMR (DMSO-d.sub.6, 300
MHz) .delta. 10.71 (s, 1H), 10.55 (s, 1H), 8.17 (s, 2H), 8.14 (s,
1H), 8.07 (dd, 1H, J=8.6, 2.4 Hz), 8.01-7.96 (m, 2H), 7.80-7.78 (m,
1H), 7.65-7.58 (m, 2H), 3.44 (br. s, 4H), 2.80 (t, 4H, J=4.8 Hz),
2.13 (s, 3H), 1.44 (s, 9H); MS (ESI) m/z 608 (M+H).sup.+.
[0549] (h) 4-{[(1-{3-[2-(6-Acetylamino-pyridin-3-yl)-ethyl]-phenyl
1-methanoyl)-amino]-trifluoro-methyl-phenyl}-piperazine-1-carboxylic
acid tert-butyl ester, AA-1 h, was prepared in a manner similar to
that described in example S-1, step (d) for
3-(2-isoquinolin-4yl-ethyl)-benzoi- c acid ethyl ester, S-1d,
except 4-[({1-[3-(6-acetylamino-pyridin-3-ylethy-
nyl)-phenyl]-methanoyl}-amino)-trifluoromethyl-phenyl]-piperazine-1-carbox-
ylic acid tert-butyl ester, AA-1 g, was used instead of ethyl
3-(isoquinolin-4-ylethynyl)benzoate and the reduction was done at
45 psi of hydrogen in acetic acid-methanol-tetrahydrofuran: HPLC R,
17.1 min.; TLC R.sub.f 0.6 (4% methanol-dichloromethane); .sup.1H
NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.40 (s, 1H), 10.35 (s, 1H),
8.16-8.14 (m, 2H), 8.05 (dd, 1H, J=8.8, 2.4 Hz), 7.98 (d, 1H, J=8.3
Hz), 7.84 (s, 1H), 7.81-7.77 (m, 1H), 7.64 (dd, 1H, J=8.6, 2.4 Hz),
7.58 (d, 1H, J=8.8 Hz), 7.44 (d, 1H, J=4.8 Hz), 3.44 (br. s, 4H),
2.98-2.91 (m, 4H), 2.80 (t, 4H, J=4.6 Hz), 2.06 (s, 3H), 1.43 (s,
9H); MS (ESI) m/z 612 (M+H).sup.+.
[0550] (i)
3-[2-(6-Acetylamino-pyridin-3-yl)-ethyl]-N-(4-piperazin-1-yl-3--
trifluoromethyl-phenyl)-benzamide dihydrochloride, AA-1, was
prepared in a manner similar to that described in example R-13,
step (b) for
4-fluoro-N-[4-(piperazin-1-yl)-3-trifluoromethylphenyl]-3-(pyridin-3-yl)m-
ethoxybenzamide, R-13, except 4-{[(1
{3-[2-(6-acetylamino-pyridin-3-yl)-et-
hyl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carbox-
ylic acid tert-butyl ester, AA-1h, was used in place of
4-fluoro-N-[4-{t-butoxycarbonyl)piperazin-1-yl}-3-trifluoromethylphenyl]--
3-(pyridin-3-yl)methoxybenzamide, R-13a, and the deprotection was
done with HCl in ethanol instead of trifluoroacetic acid in
methylene chloride: HPLC R.sub.1 11.7 min.; TLC R.sub.f 0.4 (15%
methanol-chloroform w/ 0.1% ammonium hydroxide); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 10.97 (s, 1H), 10.52 (s, 1H), 9.23
(m, 2H), 8.22-8.12 (m, 3H), 7.91-7.82 (m, 4H), 7.56 (d, 1H, J=8.8
Hz), 7.44 (d, 2H, J=4.7 Hz), 3.17 (br. s, 4H), 3.07 (br. s, 4H),
2.97 (s, 4H), 2.12 (s, 3H); MS (ESI) m/z 512 (M+H).sup.+. Anal.
calcd for C.sub.27H.sub.28F.sub.3N.sub.5O.sub.2x
2.0HClx1.2H.sub.2O: C, 53.50; H, 5.39; N, 11.56; Cl, 11.70. Found:
C, 53.44; H, 5.54; N, 11.19; Cl, 11.62.
EXAMPLE AA-2
3-[2-(6-Amino-pyridin-3-yl)-ethyl]-N-(4-piperazin-1-yl-3-trifluoromethyl-p-
henyl)-benzamide Dihydrochloride
[0551] 346
[0552] (a) To a solution of
4-{[(1-{3-[2-(6-acetylamino-pyridin-3-yl)-ethy-
l]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxyl-
ic acid tert-butyl ester, AA-1h, (100 mg, 0.16 mmol, 1.0 eq) in
ethanol (3 mL) was added aqueous sodium hydroxide (2.8 mL, 1.0 M,
2.8 mmol, 1.8 eq). The clear solution was heated to 55.degree. C.
for 18 hours and poured into a mixture of aqueous 50% saturated
sodium bicarbonate and ethyl acetate. The organic layer was washed
with brine, dried over magnesium sulfate, filtered and purified by
radial chromatography over silica, which was eluted with 3%
methanol-chloroform with 0.1% ammonium hydroxide, to give
4-{[(1-{3-[2-(6-amino-pyridin-3-yl)-ethyl]-phenyl}-met-
hanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic acid
tert-butyl ester, AA-2a, as a white solid (54 mg, 59%): HPLC
R.sub.t 17.0 min.; TLC R.sub.f 0.5 (5% methanol-chloroform w/ 0.1%
ammonium hydroxide); .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta.
10.39 (s, 1H), 8.16 (d, 1H, J=2.4 Hz), 8.05 (dd, 1H, J=8.6, 2.2
Hz), 7.82-7.76 (m, 2H), 7.74 (dd, 1H, J=2.0 Hz), 7.57 (d, 1H, J=8.8
Hz), 7.44-7.42 (m, 2H), 7.27 (dd, 1H, J=8.4, 2.4 Hz), 6.38 (d, 1H,
J=8.4 Hz), 5.63 (s, 2H), 3.44 (br. s, 4H), 2.92-2.72 (s, 8H), 1.43
(s, 9H); MS (ESI) m/z 570 (M+H).sup.+.
[0553] (b)
3-[2-(6-Amino-pyridin-3-yl)-ethyl]-N-(4-piperazin-1-yl-3-triflu-
oromethyl-phenyl)-benzamide dihydrochloride, AA-2, was prepared in
a manner similar to that described in example AA-1, step (i),
except
4-{[(1-{3-[2-(6-amino-pyridin-3-yl)-ethyl]-phenyl}-methanoyl)-amino]-trif-
luoromethyl-phenyl}-piperazine-1-carboxylic acid tert-butyl ester,
AA-2a, was used in place of
4-{[(1-{3[2-(6-acetylamino-pyridin-3-yl)-ethyl]-phen-
yl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic
acid tert-butyl ester, AA-1h: HPLC R.sub.t 11.2 min.; .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 13.89 (s, 1H), 10.59 (s, 1H), 9.33
(s, 2H), 8.23 (d, 1H, J=2.2 Hz), 8.15-8.13 (m, 1H), 7.95-7.79 (m,
6H), 7.54 (d, 1H, J=8.7 Hz), 7.45-7.43 (m, 2H), 6.96 (d, 1H, J=9.0
Hz), 3.17 (s, 4H), 3.08 (s, 4H), 2.94-2.93 (m, 2H), 2.88-2.87 (m,
2H); MS (ESI) m/z 470 (M+H).sup.+. Anal. calcd for
C.sub.25H.sub.26F.sub.3N.sub.5Ox2.0 HCl: C, 55.36; H, 5.20; N,
12.91; Cl, 13.07. Found: C, 55.18; H, 5.16; N, 12.65; Cl,
13.28.
EXAMPLE BB-1
3-[2-(3H-Imidazo[4,5-b]pyridin-6-yl)-ethyl]-N-(4-piperazin-1-yl-3-trifluor-
omethyl-phenyl)-benzamide Dihydrochloride
[0554] 347
[0555] (a) To a solution of 5-bromo-3-nitro-pyridin-2-ylamine,
BB-1a, (Lancaster, 8.9 g, 40.8 mmol, 1.0 eq) in ethanol (450 mL)
was added tin(II) chloride dihydrate (32.3 g, 143 mmol, 3.5 eq).
The resultant yellow slurry was warmed to 60.degree. C. for 4 h.
The solvent was removed under reduce pressure and the crude
reaction mixture was treated with 10% ammonium hydroxide and
extracted with ethyl acetate. The combined organic extracts were
dried over magnesium sulfate, filtered, and concentrated under
reduced pressure to give a black solid (7.5 g). The crude product
was purified over silica (flash), which was eluted with 2-7%
methanol-ethyl acetate, to give 5-bromo-pyridine-2,3-diamine,
BB-1b, as a grey solid (7.0 g, 91%): HPLC R.sub.t 5.5 min.; TLC
R.sub.f 0.5 (1% methanol-ethyl acetate); .sup.1H NMR (DMSO-d.sub.6,
300 MHz) .delta. 7.28 (d, 1H, J=2.0 Hz), 6.80 (d, 1H, J=2.0 Hz),
5.60 (s, 2H), 4.99 (s, 2H); .sup.13C NMR (DMSO-d.sub.6, 75 MHz)
.delta. 147.4, 134.0, 131.9, 119.1, 106.4; MS im/z 188/190
(M+H).sup.+.
[0556] (b) A round bottom flask was charged with
5-bromo-pyridine-2,3-diam- ine, BB-1b, (4.6 g, 24.5 mmol) and
triethylformate (50 mL). The resultant purple slurry was warmed to
130.degree. C. for 18 h. The solvent was removed under reduced
pressure at 85.degree. C. and the resultant brown oil was dissolved
in ethanol (70 mL) and treated with concentrated HCl (30 mL). The
resultant brown slurry was warmed to 90.degree. C. for 1 hour,
concentrated under reduced pressure and treated with water (100
mL). The pH was adjusted to 9 with concentrated ammonium hydroxide
and the aqueous layer was extracted with 10% isopropyl
alcohol-ethyl acetate (5.times.100 mL). The combined organic layers
were washed with aqueous 5% sodium bicarbonate, brine, dried over
magnesium sulfate and filtered to give
6-bromo-3H-imidazo[4,5-b]pyridine, BB-1c, as a brown solid (4.5 g,
94%): HPLC R.sub.t 6.1 min.; .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 13.35, 12.93 (2 br. s, 1H), 8.56 (s, 1H), 8.50 (d, 1H,
J=1.6 Hz), 8.37 (br. s, 1H); MS (ESI) m/z 196/198 (M-H).sup.-.
Anal. calcd for C.sub.6H.sub.4BrN.sub.3: C, 36.39; H, 2.04; N,
21.22; Br, 40.35. Found: C, 36.21; H, 2.09; N, 21.11; Br,
40.28.
[0557] (c) 6-Triethylsilanylethynyl-3H-imidazo[4,5-b]pyridine,
BB-1d, was prepared in the manner similar to that described in
example AA-1, step (b), except 6-bromo-3H-imidazo[4,5-b]pyridine,
BB-1c, was used in place of N-(5-bromo-pyridin-2-yl)-acetamide,
AA-1a: HPLC R.sub.t 14.8 min.; TLC R.sub.f 0.4 (5%
methanol-methylene chloride); .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 13.34, 12.88 (2 br. s, 1H), 8.53 (s, 1H), 8.44 (s, 1H),
8.11 (br. s, 1H), 1.03 (t, 9H, J=7.8 Hz), 0.70 (q, 6H, J=7.8 Hz);
MS (ESI) m/z 258 (M+H).sup.+.
[0558] (d) To a solution of
6-triethylsilanylethynyl-3H-imidazo[4,5-b]pyri- dine, BB-1d, (1.4
g, 5.4 mmol, 1.0 eq) in methanol (30 mL) was added aqueous sodium
hydroxide (2.2 mL, 10 M, 22 mmol, 4.0 eq). The resultant brown
solution was warmed to 40.degree. C. for 24 h. The reaction mixture
was poured into aqueous 5% sodium bicarbonate (200 mL) and the
aqueous layer was extracted with 10% isopropyl alcohol-chloroform.
The organic layer was washed with brine, dried over magnesium
sulfate, filtered and concentrated under reduced pressure to give a
yellow solid (1.3 g). The crude product was washed with
methyl-tert-butyl ether/cyclohexane (2:1) to give
6-ethynyl-3H-imidazo[4,5-b]pyridine, BB-1e, as a yellow solid (750
mg, 97%): HPLC R.sub.t 5.1 min.; TLC R.sub.f 0.5 (5%
methanol-chloroform w/ 0.1% ammonium hydroxide); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 13.16 (br. s, 1H), 8.52 (s, 1H),
8.46 (d, 1H, J.=1.7 Hz), 8.14 (s, 1H), 4.26 (s, 1H); MS (ESI) m/z
144 (M+H).sup.+.
[0559] (e)
4-[({1-[3-(3H-Imidazo[4,5-b]pyridin-6-ylethynyl)-phenyl]-methan-
oyl}-amino)-trifluoro-methyl-phenyl]-piperazine-1-carboxylic acid
tert-butyl ester, BB-1f, was prepared in the manner similar to that
described in example AA-1, step (g), except
6-ethynyl-3H-[4,5-b]pyridine, BB-1e, was used instead of
N-(5-ethynyl-pyridin-2-yl)-acetamide, AA-1c: HPLC R.sub.t 16.1
min.; TLC R.sub.f 0.5 (5% methanol-chloroform w/ 0.1% ammonium
hydroxide); .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 13.20 (br.
s, 1H), 10.57 (s, 1H), 8.60 (d, 1H, J=1.5 Hz), 8.56 (s, 1H),
8.26-8.18 (m, 3H), 8.08 (dd, 1H, J=8.8, 2.2 Hz), 7.83 (d, 1H, J=7.7
Hz), 7.80 (d, 1H, J=8.0 Hz), 7.66-7.59 (m, 2H), 3.44 (br. s, 4H),
2.81 (t, 4H, J=4.6 Hz), 1.44 (s, 9H); MS (ESI) m/z 591
(M+H).sup.+.
[0560] (f)
4-{[(1-{3-[2-(3H-Imidazo[4,5-b]pyridin-6-yl)-ethyl]-phenyl}-met-
hanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic acid
tert-butyl ester, BB-1g, was prepared in the manner similar to that
described in example AA-1, step (h), except
4-[({1-[3-(3H-[imidazo[4,5-b]-
pyridin-6-ylethynyl)-phenyl]-methanoyl}-amino)-trifluoromethyl-phenyl]-pip-
erazine-1-carboxylic acid tert-butyl ester, BB-1f, was used instead
of
4-{[(1-{3-[2-(6-acetylamino-pyridin-3-yl)-ethyl]-phenyl}-methanoyl)-amino-
]-trifluoromethyl-phenyl}-piperazine-1-carboxylic acid tert-butyl
ester, AA-1 g: HPLC R, 15.4 min.; .sup.1H NMR (DMSO-d.sub.6, 300
MHz) .delta. 12.93, 12.56 (2br. s, 1H), 10.40 (s, 1H), 8.35 (s,
1H), 8.31 (br. s, 1H), 8.16 (d, 1H, J=2.4 Hz), 8.05 (dd, 1H, J=8.8,
2.2 Hz), 7.87 (s, 1H), 7.80-7.78 (m, 2H), 7.58 (d, 1H, J=8.8 Hz),
7.46-7.41 (m, 2H), 3.44 (br. s, 4H), 3.11-3.03 (m, 4H), 2.80 (t,
4H, J=4.6 Hz), 1.43 (s, 9H); MS (ESI) m/z 595 (M+H).sup.+.
[0561] (g)
3-[2-(3H-Imidazo[4,5-b]pyridin-6-yl)-ethyl]-N-(4-piperazin-1-yl-
-3-trifluoromethyl-phenyl)-benzamide, BB-1, was prepared in the
manner similar to that described in example AA-1, step (i), except
4-{[(1-{3-[2-(3H-imidazo[4,5-b]pyridin-6-yl)-ethyl]-phenyl}-methanoyl)-am-
ino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic acid
tert-butyl ester, BB-1g, was used in place of
4-{[(1-{3-[2-(6-acetylamino-pyridin-3--
yl)-ethyl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1--
carboxylic acid tert-butyl ester, AA-1h: HPLC R.sub.t 10.0 min.;
.sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.57 (s, 1H), 9.28
(br. s, 3H), 8.54 (s, 1H), 8.28-8.22 (m, 2H), 8.14 (d, 1H, J=8.8
Hz), 7.95 (s, 1H), 7.83 (d, 1H, J=6.9 Hz), 7.54 (d, 1H, J=8.9 Hz),
7.47-7.44 (m, 2H), 3.18 (br. s, 6H), 3.07 (br. s, 6H); MS (ESI) m/z
495 (M+H).sup.+. Anal. calcd for
C.sub.26H.sub.25SF.sub.3N.sub.6Ox2.0 HClx1.0H.sub.2Ox0.3
CH.sub.2Cl.sub.2: C, 51.70; H, 4.88; N, 13.76; Cl, 15.09. Found: C,
51.86; H, 4.95; N, 13.32; Cl, 14.98.
EXAMPLE CC-1
5-{2-[3-(Piperazin-1-yl-trifluoromethyl-phenylcarbamoyl)-phenyl]-ethyl}-ni-
cotinamide Dihydrochloride
[0562] 348
[0563] (a) 5-Triethylsilanylethynyl-nicotinic acid methyl ester,
CC-1a, was prepared in the manner similar to that described in
example AA-1, step (b), except 5-bromo-nicotinic acid methyl ester
was used in place of N-(5-bromo-pyridin-2-yl)-acetamide, AA-1a:
HPLC R.sub.t 20.1 min.; TLC R.sub.f 0.4 (20% ethyl
acetate-cyclohexane); .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta.
9.04 (d, 1H, J=2.0 Hz), 8.88 (d, 1H, J=2.0 Hz), 8.25 (t, 1H, J=2.1
Hz), 3.90 (s, 3H), 1.02 (t, 9H, J=8.1 Hz), 0.70 (q, 6H, J=7.6
Hz).
[0564] (b) 5-Ethynyl-nicotinic acid methyl ester, CC-1b, was
prepared in the manner similar to that described in example AA-1,
step (c), except 5-triethylsilanylethynyl-nicotinic acid methyl
ester, CC-1a, was used in place of
N-(5-triethylsilanylethynyl-pyridin-2-yl)-acetamide, AA-1b: HPLC
R.sub.t 9.3 min.; TLC R.sub.f 0.2 (20% ethyl acetate-cyclohexane);
.sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 9.06 (s, 1H), 8.91 (s
1H), 8.30 (s, 1H), 4.58 (s, 1H), 3.90 (s, 3H).
[0565] (c)
4-({2-[3-(5-Methoxycarbonyl-pyridin-3-ylethynyl)-phenyl]-2-oxo--
ethyl}-trifluoromethyl-phenyl)-piperazine-1-carboxylic acid
tert-butyl ester, CC-1c, was prepared in the manner similar to that
described in example AA-1, step (g), except 5-ethynyl-nicotinic
acid methyl ester, CC-1b, was used instead of
N-(5-ethynyl-pyridin-2-yl)-acetamide, AA-1c: HPLC R.sub.t 19.5
min.; TLC R.sub.f 0.3 (30% ethyl acetate-cyclohexane); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 10.58 (s, 1H), 9.10 (d, 1H, J=2.0
Hz), 9.04 (d, 1H, J=2.0 Hz), 8.45 (t, 1H, J=2.1 Hz), 8.24 (s, 1H),
8.18 (d, 1H, J=2.3 Hz), 8.09-8.03 (m, 2H), 7.86 (d, 1H, J=7.8 Hz),
7.68-7.59 (m, 2H), 3.93 (s, 3H), 3.44 (br. s, 4H), 2.82-2.79 (m,
4H), 1.41 (s, 9H).
[0566] (d) To a solution of
4-({2-[3-(5-methoxycarbonyl-pyridin-3-ylethyny-
l)-phenyl]-2-oxo-ethyl}-trifluoromethyl-phenyl)-piperazine-1-carboxylic
acid tert-butyl ester, CC-1c, (846 mg, 1.4 mmol, 1.0 eq) in
isopropyl alcohol (90 mL) was added aqueous sodium hydroxide (4.2
mL, 1.0 M, 4.2 mmol, 3.0 eq). The resultant clear solution was
warmed to 50.degree. C. for 5 h. The solvent was removed under
reduced pressure and the crude product was dissolved in ethyl
acetate, which was sequentially washed with aqueous sodium citrate
(0.5 M, pH 4.5) and brine. The organic layer was dried over
magnesium sulfate, filtered and concentrated under reduced pressure
to give 4-({2-[3-(5-carboxy-pyridin-3-ylethynyl)-phenyl]-2-oxo-e-
thyl}-trifluoromethyl-phenyl)-piperazine-1-carboxylic acid
tert-butyl ester, CC-1d, as a white solid (800 mg, 96%): .sup.1H
NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.59 (s, 1H), 9.08 (d, 1H,
J=2.0 Hz), 9.00 (d, 1H, J=2.0 Hz), 8.42 (t, 1H, J=2.1 Hz), 8.24 (s,
1H), 8.18 (d, 1H, J=2.4 Hz), 8.09-8.02 (m, 2H), 7.88-7.85 (m, 1H),
7.68-7.59 (m, 2H), 3.44 (br. s, 4H), 2.81-2.78 (m, 4H), 1.43 (s,
9H).
[0567] (e)
4-[(2-{3-[2-(5-Carboxy-pyridin-3-yl)-ethyl]-phenyl}-2-oxo-ethyl-
)-trifluoromethyl-phenyl]-piperazine-1-carboxylic acid tert-butyl
ester, CC-1e, was prepared in the manner similar to that described
in example AA-1, step (h), except
4-({2-[3-(5-carboxy-pyridin-3-ylethynyl)-phenyl]-2-
-oxo-ethyl}-trifluoromethyl-phenyl)-piperazine-1-carboxylic acid
tert-butyl ester, CC-1d, was used instead of
4-{[(1-{3-[2-(6-acetylamino--
pyridin-3-yl)-ethyl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-pip-
erazine-1-carboxylic acid tert-butyl ester, AA-1g: HPLC (TFA
buffered method) R.sub.t 16.4 min.; TLC R.sub.f 0.3 (3%
methanol-dichloromethane w/ 0.1% acetic acid); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 10.47 (s, 1H), 8.89 (s, 1H), 8.57
(s, 1H), 8.17-8.15 (m, 2H), 8.08-8.05 (m, 1H), 7.88 (s, 1H),
7.81-7.79 (m, 1H), 7.58 (d, 1H, J=8.8 Hz), 7.46-7.44 (m, 2H), 3.01
(s, 5H), 2.79 (m, 5H), 1.43 (s, 9H); MS (ESI) m/z 597
(M-H).sup.-.
[0568] (f) To a solution of
4-[(2-{3-[2-(5-carboxy-pyridin-3-yl)-ethyl]-ph-
enyl}-2-oxo-ethyl)-trifluoromethyl-phenyl]-piperazine-1-carboxylic
acid tert-butyl ester, CC-1e, (250 mg, 0.42 mmol, 1.0 eq) in
dioxane (9 mL) was added a solution of 1,3-dicyclohexylcarbodiimide
(87 mg, 0.42 mmol, 1.0 eq) and N-hydroxysuccinimide (48 mg, 0.42
mmol, 1.0 eq) in dioxane (1 mL). The cloudy reaction mixture was
sequentially stirred for 18 h, passed through a teflon filter (0.45
micron pore size), and the solvent removed under reduced pressure
to give a white semi-solid (346 mg). To the crude product in
dioxane (8 mL) was added triethylamine (0.23 mL, 1.68 mmol, 4.0 eq)
and 1,2-diaminomethane dihydrochloride (50 mg, 0.42 mmol, 1.0 eq)
in a minimal amount of water. The white slurry was stirred for 4 h
at room temperature. The solvent was removed under reduced pressure
and the crude product dissolved in ethyl acetate, which was
sequentially washed with water and brine. The organic layer was
dried over magnesium sulfate, filtered and concentrated under
reduced pressure to give a yellow solid (360 mg). The crude product
was purified by radial chromatography over silica gel, which was
eluted with 3-9% methanol-dichloromethane, to give
4-[(2-{3-[2-(5-carbamoyl-pyridin-3-yl)--
ethyl]-phenyl}-2-oxo-ethyl)-trifluoromethyl-phenyl]-piperazine-1-carboxyli-
c acid tert-butyl ester, CC-1f, as a white solid (150 mg, 60%):
HPLC R.sub.t 15.4 min.; TLC R.sub.f 0.5 (4%
methanol-dichloromethane); .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 10.45 (s, 1H), 8.68 (d, 1H, J=1.8 Hz), 8.56 (d, 1H, J=1.8
Hz), 8.17-8.04 (m, 3H), 7.87-7.79 (m, 1H), 7.61-7.58 (m, 2H),
7.47-7.45 (m, 2H), 5.76 (s, 2H), 3.44 (s, 4H), 3.02 (m, 4H), 2.80
(s, 4H), 1.43 (s, 9H); MS (ESI) m/z 598 (M+H).sup.+.
[0569] (g)
5-(2-{3-[2-(Piperazin-1-yl-trifluoromethyl-phenyl)-ethanoyl]-ph-
enyl}-ethyl)-nicotinamide dihydrochloride, CC-1, was prepared in
the manner similar to that described in example AA-1, step (i),
except
4-[(2-{3-[2-(5-carbamoyl-pyridin-3-yl)-ethyl]-phenyl}-2-oxo-ethyl)-triflu-
oromethyl-phenyl]-piperazine-1-carboxylic acid tert-butyl ester,
CC-1f, was used in place of
4-{[(1-{3-[2-(6-acetylamino-pyridin-3-yl)-ethyl]-phe-
nyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic
acid tert-butyl ester, AA-1h: HPLC R.sub.t 9.7 min.; .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 10.56 (s, 1H), 9.13 (br. s, 1H),
9.00 (d, 1H, J=2.0 Hz), 8.75 (d, 1H, J=1.9 Hz), 8.49 (s, 1H), 8.33
(s, 1H), 8.22 (d, 1H, J=2.5 Hz), 8.13 (dd, 1H, J=8.8, 2.2 Hz), 7.91
(s, 1H), 7.83-7.79 (m, 2H), 7.56 (d, 1H, J=8.7 Hz), 7.48-7.45 (m,
2H), 3.13 (s, 4H), 3.07 (m, 8H); MS (ESI) m/z 498 (M+H).sup.+.
Anal. calcd for C.sub.26H.sub.26F.sub.3N.sub.5O.sub.2x 2.0
HClx0.5H.sub.2O: C, 53.89; H, 5.04; N, 12.09; Cl, 12.24. Found: C,
53.99; H, 5.19; N, 11.46; Cl, 11.79.
EXAMPLE CC-2
5-{2-[3-(Piperazin-1-yl-trifluoromethyl-phenylcarbamoyl)-phenyl]-ethyl}-Ni-
cotinic Acid Methyl Ester Dihydrochloride
[0570] 349
[0571] (a) To a solution of
4-[(2-{3-[2-(5-carboxy-pyridin-3-yl)-ethyl]-ph-
enyl}-2-oxo-ethyl)-trifluoromethyl-phenyl]-piperazine-1-carboxylic
acid tert-butyl ester, CC-1e, (100 mg, 0.17 mmol, 1.0 eq.) in
methanol (4 mL) and toluene (4 mL) was added a solution of
(trimethylsilyl)diazomethane (0.45 mL, 2.0 M, 0.90 mmol, 5.3 eq).
The light yellow solution was stirred for 18 h. The solvent was
removed under reduced pressure and the crude product was dissolved
in ethyl acetate, which was sequentially washed with aqueous 5%
sodium bicarbonate and brine. The organic layer was dried over
magnesium sulfate, filtered and concentrated under reduced pressure
to give a clear residue (105 mg). The crude product was purified by
radial chromatography over silica gel, which was eluted with 2%
methanol-dichloromethane, to give
4-{[(1-{3-[2-(5-methoxycarbonyl-pyridin-
-3-yl)-ethyl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-
-1-carboxylic acid tert-butyl ester, CC-2a, as a white solid (74
mg, 71%): HPLC R.sub.t 18.1 min.; TLC R.sub.f 0.6 (3%
methanol-dichloromethane w/ 0.1% acetic acid); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 10.42 (s, 1H), 8.92 (d, 1H, J=1.8
Hz), 8.69 (d, 1H, J=2.0 Hz), 8.21 (s, 1H), 8.16 (d, 1H, J=2.2 Hz),
8.07-8.04 (m, 1H), 7.86 (s, 1H), 7.81-7.79 (m, 1H), 7.60 (d, 1H,
J=8.2 Hz), 7.48-7.45 (m, 2H), 3.88 (s, 3H), 3.44 (br. s, 4H), 3.04
(s, 4H), 2.80 (t, 4H, J=4.3 Hz), 1.43 (s, 9H); MS (ESI) m/z 613
(M+H).sup.+.
[0572] (b)
5-{2-[3-(Piperazin-1-yl-trifluoromethyl-phenylcarbamoyl)-phenyl-
]-ethyl}-nicotinic acid methyl ester dihydrochloride, CC-2, was
prepared in the manner similar to that described in example AA-1,
step (i), except,
4-{[(1-{3-[2-(5-methoxycarbonyl-pyridin-3-yl)-ethyl]-phenyl}-meth-
anoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic acid
tert-butyl ester, CC-2a, was used in place of
4-{[(1-{3-[2-(6-acetylamino-
-pyridin-3-yl)-ethyl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-pi-
perazine-1-carboxylic acid tert-butyl ester, AA-1h: HPLC R.sub.t
12.7 min.; .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.58 (s,
1H), 9.33 (br. s, 2H), 9.00 (s, 1H), 8.80 (s, 1H), 8.38 (s, 1H),
8.22 (s, 1H), 8.14 (d, 1H, J=8.4 Hz), 7.92 (s, 1H), 7.84-7.82 (m,
1H), 7.56 (d, 1H, J=8.6 Hz), 7.48-7.42 (m, 2H), 3.90 (s, 3H), 3.16
(s, 4H), 3.08 (s, 8H); MS (ESI) m/z 513 (M+H).sup.+. Anal. calcd
for C.sub.27H.sub.27F.sub.3N.sub.4- O.sub.3x2.0HClx0.5H.sub.2Ox 0.5
ethyl acetate: C, 54.55; H, 5.37; N, 8.77. Found: C, 54.47; H,
5.45; N, 8.71.
EXAMPLE DD-1
4-Fluoro-3-[2-(3H-imidazo[4,5-b]pyridin-6-yl)-ethyl]-N-(4-piperazin-1-yl-3-
-trifluoromethyl-phenyl)-benzamide Dihydrochloride
[0573] 350
[0574] (a)
4-({[1-(4-Fluoro-3-hydroxy-phenyl)-methanoyl]-amino}-trifluorom-
ethyl-phenyl)-piperazine-1-carboxylic acid tert-butyl ester, DD-1a,
was prepared in a similar manner to that described in example M-1,
step (e), except
4-({[1-(3-acetoxy-4-fluoro-phenyl)-methanoyl]-amino}-trifluorometh-
yl-phenyl)-piperazine-1-carboxylic acid tert-butyl ester was used
instead of acetic acid 3-(2-methyl-quinolin-6-ylcarbamoyl)-phenyl
ester, M-1d: HPLC R.sub.t 16.2 min.; TLC R.sub.f 0.3 (30% ethyl
acetate-cyclohexane); .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta.
10.37 (s, 1H), 10.22 (s, 1H), 8.14 (d, 2H, J=2.4 Hz), 8.00 (dd, 1H,
J=8.6, 2.0 Hz), 7.58-7.53 (m, 3H), 7.47-7.42 (m, 1H), 7.30 (dd, 1H,
J=11.0, 8.6 Hz), 3.43 (br. s, 4H), 2.79 (t, 4H, J=4.8 Hz), 1.42 (s,
9H); MS (ESI) m/z 484 (M+H).sup.+.
[0575] (b) To a solution of
4-({[1-(4-fluoro-3-hydroxy-phenyl)-methanoyl]--
amino}-trifluoromethyl-phenyl)-piperazine-1-carboxylic acid
tert-butyl ester, DD-1a, (900 mg, 1.86 mmol, 1.0 eq), in dioxane
(40 mL) was added triethylamine (0.90 mL, 6.5 mmol, 3.0 eq) and
1,1,1-trifluoro-N-phenyl-N--
[(trifluoromethyl)sulfonyl]methanesulfonamide (1.1 g, 3.0 mmol, 1.4
eq). The clear solution was stirred for 18 h. The solvent was
removed under reduced pressure and the resultant oil was passed
through a silica plug, which was eluted with ethyl acetate to give
an amber oil (2.0 g). The crude product was purified by radial
chromatography over silica gel, which was eluted with 15-25% ethyl
acetate-cyclohexane, to give
4-({[1-(4-fluoro-3-trifluoromethanesulfonyloxy-phenyl)-methanoyl]-amino}--
trifluoromethyl-phenyl)-piperazine-1-carboxylic acid tert-butyl
ester, DD-1b, as a white solid (1.0 g, 91%): HPLC R.sub.t 19.9
min.; TLC R.sub.f 0.5 (30% ethyl acetate-cyclohexane); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 10.61 (s, 1H), 8.25-8.22 (m, 1H),
8.12 (d, 1H, J=2.4 Hz), 8.02 (dd, 1H, J=8.8, 2.2 Hz), 7.86-7.80 (m,
1H), 7.60 (d, 1H, J=8.7 Hz), 7.30-7.25 (m, 1H), 3.44 (br. s, 4H),
2.80 (t, 4H, J=4.7 Hz), 1.43 (s, 9H); MS (ESI) m/z 638
(M+Na).sup.+.
[0576] (c)
4-[({1-[4-Fluoro-3-(3H-imidazo[4,5-b]pyridin-6-ylethynyl)-pheny-
l]-methanoyl}-amino)-trifluoromethyl-phenyl]-piperazine-1-carboxylic
acid tert-butyl ester, DD-1c, was prepared in the manner similar to
that described in example AA-1, step (g), except
6-ethynyl-3H-[4,5-b]pyridine, BB-1e, was used instead of
N-(5-ethynyl-pyridin-2-yl)-acetamide, AA-1 c, and
4-({[1-(4-fluoro-3-trifluoromethanesulfonyloxy-phenyl)-methanoyl]-ami-
no}-trifluoromethyl-phenyl)-piperazine-1-carboxylic acid tert-butyl
ester, DD-1b, was used instead of
4-({[1-(3-iodo-phenyl)-methanoyl]-amino}-trifl-
uoromethyl-phenyl)-piperazine-1-carboxylic acid tert-butyl ester,
AA-1f: HPLC R.sub.t 16.3 min.; TLC R.sub.f 0.3 (4%
methanol-chloroform w/ 0.1% ammonium hydroxide); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 13.39, 12.93 (2 br. s, 1H), 10.56
(s, 1H), 8.58-8.55 (m, 3H), 8.33 (s, 1H), 8.16 (s, 1H), 8.07-8.05
(m, 2H), 7.61-7.52 (m, 2H), 3.44 (br. s, 4H), 2.80 (br. s, 4H),
1.43 (s, 9H); MS (ESI) m/z 609 (M+H).sup.+.
[0577] (d)
4-{[(1-{4-Fluoro-3-[2-(3H-imidazo[4,5-b]pyridin-6-yl)-ethyl]-ph-
enyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic
acid tert-butyl ester, DD-1d, was prepared in the manner similar to
that described in example AA-1, step (h), except
4-[({1-[4-fluoro-3-(3H-imidaz-
o[4,5-b]pyridin-6-ylethynyl)-phenyl]-methanoyl}-amino)-trifluoromethyl-phe-
nyl]-piperazine-1-carboxylic acid tert-butyl ester, DD-1c, was used
instead of
4-{[(1-{3-[2-(6-acetylamino-pyridin-3-yl)-ethyl]-phenyl}-metha-
noyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic acid
tert-butyl ester, AA-1 g: HPLC R.sub.t 15.7 min.; TLC R.sub.f 0.4
(6% methanol-chloroform w/ 0.1% ammonium hydroxide); .sup.1H NMR
(DMSO-d.sub.6, 300 MHz) .delta. 10.43 (s, 1H), 8.37 (s, 1H), 8.21
(s, 1H), 8.13 (d, 1H, J=8.8 Hz), 8.05-7.97 (m, 2H), 7.91-7.86 (m,
2H), 7.60 (d, 1H, J=8.8 Hz), 7.34-7.28 (m, 1H), 3.44 (br. s, 4H),
3.08-3.06 (m, 4H), 2.80 (t, 4H, J=4.6 Hz), 1.43 (s, 9H); MS (ESI)
m/z 613 (M+H).sup.+.
[0578] (e)
4-Fluoro-3-[2-(3H-imidazo[4,5-b]pyridin-6-yl)-ethyl]-N-(4-piper-
azin-1-yl-3-trifluoromethyl-phenyl)-benzamide dihydrochloride,
DD-1, was prepared in the manner similar to that described in
example AA-1, step (i), except
4-{[(1-{4-fluoro-3-[2-(3H-[4,5-b]pyridin-6-yl)-ethyl]-phenyl}-
-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1-carboxylic
acid tert-butyl ester, DD-1d, was used in place of
4-{[(1-{3-[2-(6-acetylamino-
-pyridin-3-yl)-ethyl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-pi-
perazine-1-carboxylic acid tert-butyl ester, AA-1h: HPLC R, 10.5
min.; .sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.68 (s, 1H),
9.36 (br. s, 3H), 8.55 (s, 1H), 8.26 (d, 2H, J=9.8 Hz), 8.12 (t,
2H, J=8.2 Hz), 7.93 (br. s, 1H), 7.56 (d, 1H, J=8.7 Hz), 7.30 (t,
1H, J=9.1 Hz), 3.16 (br. s, 6H), 3.08 (m, 6H); MS (ESI) m/z 513
(M+H).sup.+. Anal. calcd for C.sub.26H.sub.24F.sub.4N.sub.6Ox2.0
HCl: C, 52.14; H, 4.63; N, 14.03; Cl, 11.84. Found: C, 52.54; H,
4.73; N, 13.41; Cl, 11.44.
EXAMPLE EE-1
4-Fluoro-3-(5-furan-2-yl-pyridin-3-ylmethoxy)-N-(4-piperazin-1-yl-3-triflu-
oromethyl-phenyl)-benzamide Dihydrochloride
[0579] 351
[0580] (a) To a solution of
4-[({1-[3-(5-bromo-pyridin-3-ylmethoxy)-4-fluo-
ro-phenyl]-methanoyl}-amino)-trifluoromethyl-phenyl]-piperazine-1-carboxyl-
ic acid tert-butyl ester, R-25c, (80 mg, 0.12 mmol, 1.0 eq) in
dioxane (3 mL) was added tributyl-furan-2-yl-stannane (0.05 mL,
0.14 mmol, 1.2 eq) and tetrakis(triphenylphosphine)palladium(0) (14
mg, 0.01 mmol, 10 mol %). The light yellow solution was warmed to
95.degree. C. for 18 h. The solvent was removed under reduced
pressure and the crude product was dissolved in ethyl acetate,
which was sequentially washed with aqueous 10% potassium fluoride,
water and brine. The organic layer was dried over magnesium
sulfate, filtered and concentrated under reduced pressure to give a
clear oil (127 mg). The crude product was purified by radial
chromatography over silica gel, which was eluted with 1-3%
methanol-dichloromethane, to give
4-[({1-[4-fluoro-3-(5-furan-2-yl-pyridi-
n-3-ylmethoxy)-phenyl]-methanoyl}-amino)-trifluoromethyl-phenyl]-piperazin-
e-1-carboxylic acid tert-butyl ester, EE-1a, as a white solid (66
mg, 86%): HPLC R.sub.t 18.9 min.; TLC R.sub.f 0.4 (2%
methanol-dichloromethan- e); .sup.1H NMR (DMSO-d.sub.6, 300 MHz)
.delta. 10.44 (s, 1H), 8.96 (d, 1H, J=2.1 Hz), 8.62 (d, 1H, J=1.9
Hz), 8.19 (t, 1H, J=2.0 Hz), 8.14 (d, 1H, J=2.3 Hz), 8.03 (dd, 1H,
J=8.6, 2.2 Hz), 7.89 (dd, 1H, J=8.3, 1.9 Hz), 7.86 (d, 1H, J=1.6
Hz), 7.69-7.65 (m, 1H), 7.60 (d, 1H, J=8.8 Hz), 7.44 (dd, 1H,
J=11.0, 8.6 Hz), 7.16 (d, 1H, J=3.3 Hz), 6.67 (dd, 1H, J=3.4, 1.8
Hz), 5.38 (s, 2H), 3.44 (br. s, 4H), 2.80 (t, 4H, J=4.7 Hz), 1.43
(s, 9H); MS (ESI) m/z 641 (M+H).sup.+.
[0581] (b)
4-Fluoro-3-(5-furan-2-yl-pyridin-3-ylmethoxy)-N-(4-piperazin-1--
yl-3-trifluoromethyl-phenyl)-benzamide dihydrochloride, EE-1, was
prepared in the manner similar to that described in example AA-1,
step (i), except
4-[({1-[4-fluoro-3-(5-furan-2-yl-pyridin-3-ylmethoxy)-phenyl]-methanoyl}--
amino)-trifluoromethyl-phenyl]-piperazine-1-carboxylic acid
tert-butyl ester, EE-1a, was used in place of
4-{[(1-{3-[2-(6-acetylamino-pyridin-3--
yl)-ethyl]-phenyl}-methanoyl)-amino]-trifluoromethyl-phenyl}-piperazine-1--
carboxylic acid tert-butyl ester, AA-1 h: HPLC R.sub.t 14.2 min.;
.sup.1H NMR (DMSO-d.sub.6, 300 MHz) .delta. 10.66 (s, 1H), 9.16
(br. s, 2H), 9.03 (s, 1H), 8.70 (s, 1H), 8.37 (s, 1H), 8.21 (d, 1H,
J=2.0 Hz), 8.14 (d, 1H, J=8.9 Hz), 7.99 (d, 1H, J=7.0 Hz), 7.89 (s,
1H), 7.72-7.69 (m, 1H), 7.56 (d, 1H, J=8.5 Hz), 7.44 (dd, 1H,
J=10.8, 8.7 Hz), 7.24 (d, 1H, J=3.3 Hz), 6.69 (d, 1H, J=1.5 Hz),
5.45 (s, 2H), 3.17 (s, 4H), 3.07 (s, 4H); MS (ESI) m/z 541
(M+H).sup.+. Anal. calcd for C.sub.28H.sub.24F.sub.4N.sub.4-
O.sub.3x2.0HClx1.0H.sub.2O: C, 53.26; H, 4.47; N, 8.87; Cl, 11.23.
Found: C, 53.27; H, 4.25; N, 8.56; Cl, 11.48.
[0582] The exemplary compounds described above may be tested for
their activity as described below.
[0583] Biological Testing: Enzyme Assays
[0584] The stimulation of cell proliferation by growth factors such
as VEFG, FGF, and others is dependent upon their induction of
autophosphorylation of each of their respective receptor's tyrosine
kinases. Therefore, the ability of a protein kinase inhibitor to
block cellular proliferation induced by these growth factors is
directly correlated with its ability to block receptor
autophosphorylation. To measure the protein kinase inhibition
activity of the compounds, the following constructs were
devised.
[0585] VEGF-R2 Construct for Assay: A construct (VEGF-R2.DELTA.50)
of the cytosolic domain of human vascular endothelial growth factor
receptor 2 (VEGF-R2) lacking the 50 central residues of the 68
residues of the kinase insert domain was expressed in a
baculovirus/insect cell system. Of the 1356 residues of full-length
VEGF-R2, VEGF-R2.DELTA.50 contains residues 806-939 and 990-1171,
and also one point mutation (E990V) within the kinase insert domain
relative to wild-type VEGF-R2. See commonly assigned, co-pending
U.S. patent application Ser. No. 09/390,326, filed Sep. 7, 1999,
incorporated by reference herein, for discussion of VEGF constructs
and expression systems. Autophosphorylation of the purified
construct was performed by incubation of the enzyme at a
concentration of 4 .mu.M in the presence of 3 mM ATP and 40 mM
MgCl.sub.2 in 100 mM Hepes, pH 7.5, containing 5% glycerol and 5 mM
DTT, at 4.degree. C. for 2 h. After autophosphorylation, this
construct has been shown to possess catalytic activity essentially
equivalent to the wild-type autophosphorylated kinase domain
construct. See Parast et al., Biochemistry, 37, 16788-16801
(1998).
[0586] FGF-R1 Construct for Assay: The intracellular kinase domain
of human FGF-R1 was expressed using the baculovirus vector
expression system starting from the endogenous methionine residue
456 to glutamate 766, according to the residue numbering system of
Mohammadi et al., Mol. Cell. Biol., 16, 977-989 (1996). In
addition, the construct also has the following 3 amino acid
substitutions: MA57V, C488A, and C584S.
[0587] LCK Construct for Assay: The LCK tyrosine kinase was
expressed in insect cells as an N-terminal deletion starting from
amino acid residue 223 to the end of the protein at residue 509,
with the following two amino acid substitutions at the N-terminus:
P233M and C224D.
[0588] CHK-1 Construct for Assay: C-terminally His-tagged
full-length human CHK-1 (FL-CHK-1) was expressed using the
baculovirus/insect cell system. It contains 6 histidine residues
(6xHis-tag) at the C-terminus of the 476 amino acid human CHK-1.
The protein was purified by conventional chromatographic
techniques.
[0589] Catalytically active truncations of CHK-1 may be exchanged
for the full length CHK-1 protein. A preferred truncation comprises
the kinase domain of CHK-1, which begins between amino acid
residues 1 and 16 and terminates between amino acid residues 265
and 291. See commonly assigned, co-pending U.S. patent application
Ser. No. 09/460,421, filed Dec. 14, 1999, incorporated by reference
herein, for discussion of such alternate CHK-1 constructs and
expression systems.
[0590] CDK2/Cyclin A Construct for Assay: CDK2 was purified using
published methodology (Rosenblatt et al., J. Mol. Biol., 230,
1317-1319 (1993)) from insect cells that had been infected with a
baculovirus expression vector. Cyclin A was purified from E. coli
cells expressing full-length recombinant cyclin A, and a truncated
cyclin A construct was generated by limited proteolysis and
purified as described previously (Jeffrey et al., Nature, 376,
313-320 (1995)).
[0591] CDK4/Cyclin D Construct for Assay: A complex of human CDK4
and cyclin D3, or a complex of cyclin D1 and a fusion protein of
human CDK4 and glutathione-S-transferase (GST-CDK4), was purified
using traditional biochemical chromatographic techniques from
insect cells that had been co-infected with the corresponding
baculovirus expression vectors.
[0592] TEK Construct for Assay: The intracellular kinase domain
(residues 775 to 1124, with methionine added at the N-terminus) of
human TEK/Tie-2 was expressed using the baculovirus vector
expression system. For assay purposes, the enzyme was
autophosphorylated prior to use by incubation overnight at
4.degree. C., at 10 .mu.M enzyme concentration, with 4 mM ATP, 40
mM MgCl.sub.2, and 5 mM DTT in 200 mM Hepes buffer at ph 7.5, in
90:10 water:glycerol.
[0593] VEGF-R2 Assay
[0594] Coupled Spectrophotometric (FLVK-P) Assay
[0595] The production of ADP from ATP that accompanies phosphoryl
transfer was coupled to oxidation of NADH using phosphoenolpyruvate
(PEP) and a system having pyruvate kinase (PK) and lactic
dehydrogenase (LDH). The oxidation of NADH was monitored by
following the decrease of absorbance at 340 nm (e.sub.340=6.22
cm.sup.-1 mM.sup.-1) using a Beckman DU 650 spectrophotometer.
Assay conditions for phosphorylated VEGF-R2.DELTA.50 (indicated as
FLVK-P in the tables below) were the following: 1 mM PEP; 250 .mu.M
NADH; 50 units of LDH/mL; 20 units of PK/mL; 5 mM DTT; 5.1 mM
poly(E.sub.4Y.sub.1); 1 mM ATP; and 25 mM MgCl.sub.2 in 200 mM
Hepes, pH 7.5. Assay conditions for unphosphorylated
VEGF-R2.DELTA.50 (indicated as FLVK in the tables) were the
following: 1 mM PEP; 250 uM NADH; 50 units of LDH/mL; 20 units of
PK/mL; 5 mM DTT; 20 mM poly(E.sub.4Y.sub.1); 3 mM ATP; and 60 mM
MgCl.sub.2 and 2 mM MnCl.sub.2 in 200 mM Hepes, pH 7.5. Assays were
initiated with 5 to 40 nM of enzyme. K.sub.i values were determined
by measuring enzyme activity in the presence of varying
concentrations of test compounds. The data were analyzed using
Enzyme Kinetic and Kaleidagraph software.
[0596] ELISA Assay
[0597] Formation of phosphogastrin was monitored using biotinylated
gastrin peptide (1-17) as substrate. Biotinylated phosphogastrin
was immobilized using streptavidin coated 96-well microtiter plates
followed by detection using anti-phosphotyrosine-antibody
conjugated to horseradish peroxidase. The activity of horseradish
peroxidase was monitored using 2,2'-[3-ethylbenzathiazoline
sulfonate(6)] diammonium salt (ABTS). Typical assay solutions
contained: 2 .mu.M biotinylated gastrin peptide; 5 mM DTT; 20 .mu.M
ATP; 26 mM MgCl.sub.2; and 2 mM MnCl.sub.2 in 200 mM Hepes, pH 7.5.
The assay was initiated with 0.8 nM of phosphorylated
VEGF-R2.DELTA.50. Horseradish peroxidase activity was assayed using
ABTS, 10 mM. The horseradish peroxidase reaction was quenched by
addition of acid (H.sub.2SO.sub.4), followed by absorbance reading
at 405 nm. K.sub.i values were determined by measuring enzyme
activity in the presence of varying concentrations of test
compounds. The data were analyzed using Enzyme Kinetic and
Kaleidagraph software.
[0598] FGF-R Assay
[0599] The spectrophotometric assay was carried out as described
above for VEGF-R2, except for the following changes in
concentration: FGF-R=50 nM, ATP=2 mM, and poly(E4Y1)=15 mM.
[0600] LCK Assay
[0601] The spectrophotometric assay was carried out as described
above for VEGF-R2, except for the following changes in
concentration: LCK=60 nM, MgCl.sub.2=40 nM, poly(E4Y1)=20 mM.
[0602] CHK-1 Assay
[0603] The production of ADP from ATP that accompanies phosphoryl
transfer to the synthetic substrate peptide Syntide-2
(PLARTLSVAGLPGKK) was coupled to oxidation of NADH using
phosphoenolpyruvate (PEP) through the actions of pyruvate kinase
(PK) and lactic dehydrogenase (LDH). The oxidation of NADH was
monitored by following the decrease of absorbance at 340 nm
(.di-elect cons.340=6.22 cm.sup.-1 mM.sup.-1) using a HP8452
spectrophotometer. Typical reaction solutions contained: 4 mN PEP;
0.15 mM NADH; 28 units of LDH/ml; 16 units of PK/ml; 3 mM DTT;
0.125 mM Syntide-2; 0.15 mM ATP; 25 mM MgCl.sub.2 in 50 mM TRIS, pH
7.5; and 400 mM NaCl. Assays were initiated with 10 nM of FL-CHK-1.
K.sub.i values were determined by measuring initial enzyme activity
in the presence of varying concentrations of test compounds. The
data were analyzed using Enzyme Kinetic and Kaleidagraph
software.
[0604] CDK2/Cyclin A and CDK4/Cyclin D Assays
[0605] Cyclin-dependent kinase activity was measured by quantifying
the enzyme-catalyzed, time-dependent incorporation of radioactive
phosphate from [.sup.32P]ATP into a recombinant fragment of the
retinoblastoma protein. Unless noted otherwise, assays were
performed in 96-well plates in a total volume of 50 .mu.L, in the
presence of 10 mM HEPES
(N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid]) (pH 7.4),
10 mM MgCl.sub.2, 25 .mu.M adenosine triphosphate (ATP), 1 mg/mL
ovalbumin, 5 .mu.g/.mu.L leupeptin, 1 mM dithiothreitol, 10 mM
.beta.-glycerophosphate- , 0.1 mM sodium vanadate, 1 mM sodium
fluoride, 2.5 mM ethylene glycol-bis(.beta.-aminoethyl
ether)-N,N,N'N'-tetraacetic acid (EGTA), 2% (v/v)
dimethylsulfoxide, and 0.03-0.2 .mu.Ci [.sup.32P]ATP. The substrate
(0.3-0.5 .mu.g) was purified recombinant retinoblastoma protein
fragment (Rb) (residues 386-928 of the native retinoblastoma
protein; 62.3 kDa, containing the majority of the phosphorylation
sites found in the native 106-kDa protein, as well as a tag of six
histidine residues for ease of purification). Reactions were
initiated with CDK2 (150 nM CDK2/Cyclin A complex) or CDK4 (50 nM
CDK4/Cyclin D3 complex), incubated at 30.degree. C., and terminated
after 20 minutes by the addition of ethylenediaminetetraacetic acid
(EDTA) to 250 mM. The phosphorylated substrate was then captured on
a nitrocellulose membrane using a 96-well filtration manifold, and
unincorporated radioactivity was removed by repeated washing with
0.85% phosphoric acid. Radioactivity was quantified by exposing the
dried nitrocellulose membranes to a phosphorimager. Apparent
K.sub.i values were measured by assaying enzyme activity in the
presence of different compound concentrations and subtracting the
background radioactivity measured in the absence of enzyme. The
kinetic parameters (kcat, Km for ATP) were measured for each enzyme
under the usual assay conditions by determining the dependence of
initial rates on ATP concentration. The data were fit to an
equation for competitive inhibition using Kaleidagraph (Synergy
Software), or were fit to an equation for competitive tight-binding
inhibition using the software KineTic (BioKin, Ltd.). Measured
K.sub.i values for known inhibitors against CDK4 and CDK2 agreed
with published IC.sub.50 values. The specific activity of CDK4 was
the same whether complexed to full-length cyclin D3 or the
truncated Cyclin D3 construct; both complexes also yielded very
similar K.sub.i values for selected inhibitors.
[0606] TEK/Tie-2 Assay
[0607] The spectrophotometric assay was carried out as described
above for VEGF-R2, except for the following changes in
concentration: TEK/Tie-2=200 nM, MgCl.sub.2=40 mM, and ATP=2
mM.
[0608] HUVEC Proliferation Assay
[0609] This assay determines the ability of a test compound to
inhibit the growth factor-stimulated proliferation of human
umbilical vein endothelial cells ("HUVEC"). HUVEC cells (passage
3-4, Clonetics, Corp.) were thawed into EGM2 culture medium
(Clonetics Corp) in T75 flasks. Fresh EGM2 medium was added to the
flasks 24 hours later. Four or five days later, cells were exposed
to another culture medium (F12K medium supplemented with 10% fetal
bovine serum (FBS), 60 .mu.g/ml endothelial cell growth supplement
(ECGS), and 0.1 mg/ml heparin). Exponentially-growing HUVEC cells
were used in experiments thereafter. Ten to twelve thousand HUVEC
cells were plated in 96-well dishes in 100 .mu.l of rich, culture
medium (described above). The cells were allowed to attach for 24
hours in this medium. The medium was then removed by aspiration and
105 .mu.l of starvation media (F12K+1% FBS) was added to each well.
After 24 hours, 15 .mu.l of test agent dissolved in 1% DMSO in
starvation medium or this vehicle alone was added into each
treatment well; the final DMSO concentration was 0.1%. One hour
later, 30 .mu.l of VEGF (30 ng/ml) in starvation media was added to
all wells except those containing untreated controls; the final
VEGF concentration was 6 ng/ml. Cellular proliferation was
quantified 72 hours later by MTT dye reduction, at which time cells
were exposed for 4 hours MTT (Promega Corp.). Dye reduction was
stopped by addition of a stop solution (Promega Corp.) and
absorbance at 595 .lambda. was determined on a 96-well
spectrophotometer plate reader.
[0610] Cancer Cell Proliferation (MV522) Assay
[0611] The protocol for assessing cellular proliferation in cancer
cells is similar to that used for assessments in HUVEC cells. Two
thousand lung cancer cells (line MV522, acquired from American
Tissue Cultural Collection) were seeded in growth media (RPMI1640
medium supplemented with 2 mM glutamine and 10% FBS). Cells are
allowed to attach for 1 day prior to addition of test agents and/or
vehicles. Cells are treated simultaneously with the same test
agents used in the HUVEC assay. Cellular proliferation is
quantified by MTT dye reduction assay 72 hours after exposure to
test agents. The total length of the assay is 4 days vs. 5 for
HUVEC cells because MV522 cells are not exposed to starvation
medium.
[0612] The results of the testing of the compounds using various
assays are summarized in the table below, where a notation of "% @"
indicates the percent inhibition at the stated concentration, "NI"
indicates no inhibition, "slow-binding kinetics" indicates that
curvature in the progress curves in the enzyme assay precluded the
determination of rates, and "NT" indicates compounds "not tested"
for a particular activity.
1 Huvec + FLVK-P FLVK LCK- CDK2 CDK4 HUVEC Albumin MV522 Ki Ki P Ki
CHK-1 FGF-P Ki Ki IC50 IC50 IC50 TEK-P EX # (nM) (nM) (nM) Ki (nM)
Ki (nM) (nM) (nM) (nM) (nM) (.mu.M) (.mu.M) A-1 785 NT NT NT NT NT
NT NT NT NT NT A-2 100 .mu.M NT NT NT NT NT NT NT NT NT NT A-3 32
4.5 NT NT NT NT NT 240 NT >10 NT A-4 13000 NT NT NT NT NT NT NT
NT NT NT A-5 8.64 1 37% @ NT NT NT NT 270 NT >10 NT 5 .mu.M A-6
1050 NT NT NT NT NT NT NT NT NT NT A-7 26 NT NT NT NT NT NT >700
NT NT NT A-8 3.36 1.9 NT NT NT NT NT 740 NT >10 NT B-1 111 25 NT
NT 87,000 NT NT NT NT NT NT B-10 NI @ NT NT NT NT NT NT NT NT NT NT
300 .mu.M (ELISA) B-11 NI @ NT NT NT NT NT NT NT NT NT NT 300 .mu.M
(ELISA) B-12 28 NT NT NT NT NT NT 530 NT >10 NT B-13 NI @ NT NT
NT NT NT NT NT NT NT NT 1 mM (ELISA) B-14 NI @ NT NT NT NT NT NT NT
NT NT NT 490 .mu.M (ELISA) B-15 15.7 NT NT NT NT NT NT 120 NT
>10 NT B-16 15 NT 9% @ NT NT NT NT 370 NT >10 NT 1 .mu.M B-17
6.95 NT NT NT NT NT NT >700 NT >10 NT B-18 5.84 NT NT NT NT
NT NT 130 NT >10 NT B-19 42% @ NT NT NT NT NT NT NT NT NT NT 1
.mu.M B-2 1820 NT NT NT NT NT NT NT NT NT NT B-20 26 NT NT NT NT NT
NT NT NT NT NT B-21 208 NT NT NT NT NT NT NT NT NT NT B-22 32% @ NT
NT NT NT NT NT NT NT NT NT 1 .mu.M B-23 17% @ 1 NT NT NT NT NT NT
NT NT NT NT .mu.M B-24 37 NT NT NT NT NT NT NT NT NT NT B-25 26% @
1 NT NT NT NT NT NT NT NT NT NT .mu.M B-3 406 NT NT NT NT NT NT NT
NT NT NT B-4 35000 NT NT NT NT NT NT NT NT NT NT B-5 11000 NT NT NT
NT NT NT NT NT NT NT B-6 62% @ NT NT NT NT NT NT NT NT NT NT 10
.mu.M (ELISA) B-7 3310 NT NT NT NT NT NT NT NT NT NT B-8 NI @ NT NT
NT NT NT NT NT NT NT NT 1 mM (ELISA) B-9 17000 NT NT NT NT NT NT NT
NT NT NT C-1 NI @ NT NT NT NT NT NT NT NT NT NT 1 mM (ELISA) C-2 NI
@ NT NT NT NT NT NT NT NT NT NT 50 .mu.M (ELISA) C-3 NI @ 5 NT NT
NT NT NT NT NT NT NT NT .mu.M (ELISA) D-1 28 NT NT NT NT NT NT 520
NT >10 NT D-2 1640 NT NT NT NT NT NT NT NT NT NT D-3 133 NT NT
NT NT NT NT NT NT NT NT E-1 2.21 1.4 85% @ NT 36% at 23% @ 40% @
170 >1000 >10 NT 5 .mu.M 5 .mu.M 100 .mu.M 100 .mu.M E-2 4.79
0.95 NT NT NT >100 >100 55 NT >10 NT .mu.M .mu.M F-1 2.36
NT 22% @ NT 11% @ NT NT 710 NT >10 NT 1 .mu.M 1 .mu.M F-2 14.7
NT NT NT NT NT NT 650 NT >10 NT F-3 1.01 0.31 84% @ NT 83% @ NT
NT 10 NT >10 NT 1 .mu.M 1 .mu.M F-4 slow- slow- 19% @ NT 18% @
NT NT 63 NT 2.6 NT binding binding 1 .mu.M 1 .mu.M kinetics
kinetics F-5 NT 0.86 84% @ NT 69% @ NT NT 4.1 17 1.4 56% @ 1 .mu.M
1 .mu.M 1 .mu.M G-1 0.592 NT 90 NT 81% @ NT NT 10 435 >10 NT 1
.mu.M G-10 2.6 3 48% @ NT 33% @ NT NT 5.7 180 5.1 NT 1 .mu.M 1
.mu.M G-11 0.17 0.11 2.5 7% @ 1 19 NI @ 1 16% @ 4.4 64 5.6 NT .mu.M
.mu.M 1 .mu.M G-2 1.78 NT NT NT NT NT NT 120 NT >10 NT G-3 0.197
NT 11.2 NT 91% @ NT NT 9.8 280 >10 NT 1 .mu.M G-4 0.79 0.1 72 NT
77% @ NT NT 16 NT >10 NT 1 .mu.M G-5 1.86 0.35 69 NT 68% @ NT NT
8.4 NT >10 NT 1 .mu.M G-6 1.98 NT 65 NT 74% @ NT NT 12 59 10 NT
1 .mu.M G-7 0.65 0.25 27 NT 23 NT NT 17 190 >10 14% @ 1 .mu.M
G-8 0.73 0.035 5.8 NT 20 NT NT 12 114 8.7 62% @ 1 .mu.M G-9 NT 2.5
NT NT NT NT NT NT NT NT NT H-1 1.19 0.19 41% @ NT 83% @ NT NT 27 NT
>10 NT 1 .mu.M 1 .mu.M I-1 8.84 NT NT NT NT NT NT 220 NT >10
NT I-2 16% @ NT NT NT NT NT NT NT NT NT 1 .mu.M J-1 NT 1.6 38% @ NT
61% @ NT NT 37 NT NT NT 1 .mu.M 1 .mu.M J-2 NT 0.68 10.5 NT 57 NT
NT 26 180 >10 17% @ 1 .mu.M J-3 NT 0.92 44% @ NT 54% @ NT NT 50
NT NT NT 1 .mu.M 1 .mu.M K-1 1.46 1.6 59 NT 79% @ NT NT 40 >1000
10 NT 1 .mu.M K-10 2.19 0.23 57.7 8% @ 1 26 12% @ 17% @ 12 NT 4.6
NT .mu.M 1 .mu.M 1 .mu.M K-11 3.49 NT 82% @ NT 59% @ NT NT 37 NT
8.1 NT 1 .mu.M 1 .mu.M K-12 2.96 NT 48% @ NT 70% @ NT NT 27 NT 4.7
NT 1 .mu.M 1 .mu.M K-2 138 NT NT NT NT NT NT NT NT NT NT K-3 1.66
3.4 NT 26,000 NI @ 150 NT NT 430 NT >10 NT .mu.M K-4 64 NT NT NT
NT NT NT 380 NT >10 NT K-5 1270 NT NT NT NT NT NT NT NT NT NT
K-6 200 NT NT NT NI @ NT NT NT NT NT NT 1 mM K-7 122 NT NT NT NT NT
NT NT NT NT NT K-8 707 NT NT NT NT NT NT NT NT NT NT K-9 109 10 NT
NT NI @ 600 NT NT NT NT NT NT .mu.M L-1 NT 3% @ NT NT NT NT NT NT
NT NT NT 50 nM M-1 52 NT NT NT NT NT NT NT NT NT NT N-1 3.74 1.3
160 10% @ 1 12% @ 18% @ 19% @ 38 >1000 >10 NT .mu.M 1 .mu.M 1
.mu.M 1 .mu.M N-2 NT 8% @ NT NT NT NT NT >100 NT >10 NT 50 nM
O-1 NT 5.5 NT NT NT NT NT >300 NT <3 NT O-2 NT 13% @ NT NT NT
NT NT NT NT NT NT 50 nM O-3 NT 5.8 NT NT NT NT NT NT NT NT NT P-1
4% @ 50 NT NT NT NT NT NT NT NT NT NT nM Q-1 17% @ 50 NT NT NT NT
NT NT 100-300 NT NT NT nM R-1 49% @ 50 NT NT NT NT NT NT NT NT NT
NT nM R-10 12.1 6.9 23% @ NT NT NT NT 28 NT >10 NT 1 .mu.M R-11
NT slow- 59% @ NT 30% @ 1 NT NT 31 NT NT NT binding 1 .mu.M .mu.M
59% @ 50 nM R-12 NT 4.4 45% @ NT 62% @ NT NT 15 NT 5.9 NT 1 .mu.M 1
.mu.M R-13 NT 16.5 5% @ NT 7% @ NT NT 19 120 1.8 NT 1 .mu.M 1 .mu.M
R-14 NT 3.4 87% @ NT 96% @ 1 NT NT 14 345 5.7 NT 1 .mu.M .mu.M R-15
NT 8.8 26% @ NT 41% @ 1 NT NT 36 130 3.1 NT 1 .mu.M .mu.M R-16 NT
2.3 NT NT NT NT NT 13 NT 5.6 NT R-17 NT 7.4 NT NT NT NT NT 32 NT NT
R-18 NT 3.1 NT NT NT NT NT 18 NT >10 NT R-19 NT 13.9 NT NT NT NT
NT 64 NT 1.7 NT R-2 2.53 NT NT NT NT NT NT 110 NT 0.43 NT R-20 NT
19% @ NT NT NT NT NT 155 NT 2.6 NT 50 nM R-3 67% @ 5 NT NT NT NT NT
NT 50 NT 9.7 NT .mu.M R-4 9.93 2.4 28% @ NT NT NT NT 88 NT 9.9 NT 1
.mu.M R-5 11.2 1.7 NT NT NT NT 89 NT 10 NT R-6 16% @ 5 NT NT NT NT
NT NT NT NT NT NT .mu.M R-7 325 NT NT NT NT NT NT NT NT NT NT R-8
4% @ NT NT NT NT NT NT NT NT NT NT 5 .mu.M R-9 9% @ NT NT NT NT NT
NT NT NT NT NT 1 .mu.M R-21 NT 42% @ NT NT NT NT NT NT NT NT NT 50
nM R-22 NT 52% @ 13% @ NT 21% @ 1 NT NT NT >100 NT NT 50 nM 1
.mu.M .mu.M R-23 NT 50% @ NT NT NT NT NT NT NT NT NT 50 nM R-24 NT
62% @ NT NT NT NT NT NT NT NT NT 50 nM R-25 NT 6.6 NT NT NT NT NT
NT 155 NT NT S-1 2% @ NT NT NT NT NT NT NT NT NT NT 1 .mu.M S-2 28%
@ 1 NT NT NT NT NT NT NT NT NT NT .mu.M S-3 8.7 0.76 NT NT NT NT NT
180 NT NT NT S-4 20% @ 50 NT NT NT NT NT NT >300 NT NT NT nM S-5
4.2 NT NT NT NT NT NT >300 NT NT NT S-6 6% @ 50 NT NT NT NT NT
NT NT NT NT NT nM S-7 NT 19% @ NT NT NT NT NT >300 NT NT NT 50
nM S-8 NT 1.4 NT NT NT NT NT NT NT NT NT T-1 13.6 NT NT NT NT NT NT
NT NT NT NT U-1 68 NT NT NT NT NT NT NT NT NT NT U-2 12.4 NT NT NT
NT NT NT 100 NT >10 NT V-1 NT 2 NT NT NT NT NT NT NT NT NT V-2
13 NT NT NT NT NT NT 180 NT 10 NT V-3 42% @ NT NT NT NT NT NT NT NT
NT NT 1 .mu.M V-4 NT 0.045 22% @ NT 37% @ NT NT 27 >1000 NT 40%
@ 1 .mu.M 1 .mu.M 1 .mu.M V-5 NT 1.4 NT NT NT NT NT NT NT NT NT V-6
NT 12% @ NT NT NT NT NT NT NT NT NT 50 nM V-7 NT 2.6 NT NT NT NT NT
NT NT NT NT V-8 NT 0% @ NT NT NT NT NT NT NT NT NT 50 nM V-9 NT 22
NT NT NT NT NT NT NT NT NT V-10 NT 16% @ NT NT NT NT NT NT NT NT NT
50 .mu.M V-11 NT 18% @ NT NT NT NT NT NT NT NT NT 50 nM V-12 NT 18%
@ NT NT NT NT NT NT NT NT NT 50 nM V-13 NT 17% @ NT NT NT NT NT NT
NT NT NT 50 nM V-15 NT 30% @ NT NT NT NT NT NT NT NT NT 50 nM W-1
NT 0.12 88% @ NT 67% @ NT NT 23 NT NT NT 1 .mu.M 1 .mu.M W-2 NT 1.5
19% @ NT 18% @ NT NT 120 NT NT NT 1 .mu.M 1 .mu.M W-3 NT 0.7 65% @
NT 56% @ NT NT 220 NT NT NT 1 .mu.M 1 .mu.M X-1 NT 1.5 59% @ NT 28%
@ NT NT 27 330 NT NT 1 .mu.M 1 .mu.M X-2 NT 9.1 NT NT NT NT NT NT
NT NT NT X-3 NT 2.2 NT NT NT NT NT 30-100 NT 4.6 NT X-4 NT 4.1 11%
@ NT 11% @ NT NT 71 >1000 6.1 NT 1 .mu.M 1 .mu.M X-5 NT 1.3 NT
NT NT NT NT 29 NT 3.7 NT Y-1 NT 6% @ NT NT NT NT NT >700 NT NT
NT 50 nM Y-2 NT NI @ NT NT NT NT NT NT NT NT NT 50 nM Z-1 NI @ NT
NT NT NT NT NT NT NT NT NT 5 .mu.M AA-1 59% @ 50 2.8 44% @ NT 39% @
NT NT NT 25 NT NT nM 1 .mu.M 1 .mu.M AA-2 NT 25 NT NT NT NT NT NT
80 NT NT BB-1 NT 10.8 NT NT NT NT NT NT 940 NT NT CC-1 NT 20% @ NT
NT NT NT NT NT NT NT NT 50 nM CC-2 NT 30% @ NT NT NT NT NT NT NT NT
NT 50 nM DD-1 NT 5.6 NT NT NT NT NT NT 700 NT NT EE-1 NT 3.4 53% @
NT 32% @ NT NT NT 180 NT NT 1 .mu.M 1 .mu.M
[0613] The exemplary compounds described above may be formulated
into pharmaceutical compositions according to the following general
examples.
EXAMPLE 1
Parenteral Composition
[0614] To prepare a parenteral pharmaceutical composition suitable
for administration by injection, 100 mg of a water-soluble salt of
a compound of Formula I is dissolved in DMSO and then mixed with 10
mL of 0.9% sterile saline. The mixture is incorporated into a
dosage unit form suitable for administration by injection.
EXAMPLE 2
Oral Composition
[0615] To prepare a pharmaceutical composition for oral delivery,
100 mg of a compound of Formula I is mixed with 750 mg of lactose.
The mixture is incorporated into an oral dosage unit for, such as a
hard gelatin capsule, which is suitable for oral
administration.
EXAMPLE 3
Intraocular Composition
[0616] To prepare a sustained-release pharmaceutical for
intraocular delivery, a compound of Formula I is suspended in a
neutral, isotonic solution of hyaluronic acid (1.5% conc.) in
phosphate buffer (pH 7.4) to form a 1% suspension.
[0617] It is to be understood that the foregoing description is
exemplary and explanatory in nature, and is intended to illustrate
the invention and its preferred embodiments. Thus, the scope of the
invention should be understood to be defined not by the foregoing
description, but by the following claims and their equivalents.
* * * * *