U.S. patent application number 14/996961 was filed with the patent office on 2016-05-12 for inhibitors of hepatitis c virus.
The applicant listed for this patent is Gilead Sciences, Inc.. Invention is credited to Kyla Bjornson, Eda Canales, Jeromy J. Cottell, Kapil K. Karki, Ashley A. Katana, Darryl Kato, Tetsuya Kobayashi, John O. Link, Ruben Martinez, Barton W. Phillips, Hyung-jung Pyun, Michael Sangi, Adam J. Schrier, Dustin Siegel, James G. Taylor, Chinh V. Tran, Teresa A. Trejo Martin, Randall W. Vivian, Zheng-Yu Yang, Jeff Zablocki, Sheila Zipfel.
Application Number | 20160130300 14/996961 |
Document ID | / |
Family ID | 48803614 |
Filed Date | 2016-05-12 |
United States Patent
Application |
20160130300 |
Kind Code |
A1 |
Bjornson; Kyla ; et
al. |
May 12, 2016 |
INHIBITORS OF HEPATITIS C VIRUS
Abstract
Compounds of Formula I are disclosed ##STR00001## As well as
pharmaceutically acceptable salts thereof. Methods of using said
compounds and pharmaceutical compositions containing said compounds
are also disclosed.
Inventors: |
Bjornson; Kyla; (San
Lorenzo, CA) ; Canales; Eda; (San Mateo, CA) ;
Cottell; Jeromy J.; (Foster City, CA) ; Karki; Kapil
K.; (Foster City, CA) ; Katana; Ashley A.;
(San Mateo, CA) ; Kato; Darryl; (San Francisco,
CA) ; Kobayashi; Tetsuya; (Pleasanton, CA) ;
Link; John O.; (San Francisco, CA) ; Martinez;
Ruben; (San Diego, CA) ; Phillips; Barton W.;
(San Mateo, CA) ; Pyun; Hyung-jung; (Fremont,
CA) ; Sangi; Michael; (San Mateo, CA) ;
Schrier; Adam J.; (Redwood City, CA) ; Siegel;
Dustin; (San Carlos, CA) ; Taylor; James G.;
(Foster City, CA) ; Tran; Chinh V.; (San Diego,
CA) ; Trejo Martin; Teresa A.; (Foster City, CA)
; Vivian; Randall W.; (Foster City, CA) ; Yang;
Zheng-Yu; (Palo Alto, CA) ; Zablocki; Jeff;
(Los Altos, CA) ; Zipfel; Sheila; (San Mateo,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Gilead Sciences, Inc. |
Foster City |
CA |
US |
|
|
Family ID: |
48803614 |
Appl. No.: |
14/996961 |
Filed: |
January 15, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13934090 |
Jul 2, 2013 |
9296782 |
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14996961 |
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61798524 |
Mar 15, 2013 |
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61667806 |
Jul 3, 2012 |
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Current U.S.
Class: |
530/323 |
Current CPC
Class: |
A61P 31/00 20180101;
A61K 38/21 20130101; C07K 5/08 20130101; A61P 31/10 20180101; A61K
38/00 20130101; A61K 45/06 20130101; C07K 5/0808 20130101; C07K
5/0827 20130101; A61P 31/12 20180101; A61K 31/519 20130101; C07D
498/16 20130101; A61P 43/00 20180101; A61P 1/16 20180101; A61K
31/4745 20130101; C07D 498/22 20130101; A61K 38/06 20130101; C07K
5/0812 20130101; A61K 31/4985 20130101; A61K 31/401 20130101; A61K
31/506 20130101; A61P 31/14 20180101; C07D 403/14 20130101; A61K
31/10 20130101; C07D 241/36 20130101; A61K 31/498 20130101; A61K
38/21 20130101; A61K 2300/00 20130101 |
International
Class: |
C07K 5/083 20060101
C07K005/083; C07K 5/087 20060101 C07K005/087 |
Claims
1. A compound of Formula (IV): ##STR00645## or a stereoisomer, or a
mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein: J is C.sub.1-C.sub.4 alkyl or C.sub.3-C.sub.6
carbocyclyl, wherein the C.sub.1-C.sub.4 alkyl or C.sub.3-C.sub.6
carbocyclyl is optionally substituted with 1-4 halogen, --OH, aryl
or cyano; {circle around (T)} is C.sub.3-C.sub.5 carbocyclylene
that is attached to L and to the remainder of the compound of
Formula IV through two adjacent carbons, wherein said
C.sub.3-C.sub.5 carbocyclylene is optionally substituted with
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.3 haloalkyl, halogen, --OH, or
cyano, or {circle around (T)} is C.sub.5-C.sub.8 bicyclic
carbocyclylene that is attached to L and to the remainder of the
compound of Formula IV through two adjacent carbons, or
C.sub.3--C.sub.6 carbocyclylene that is attached to L and to the
remainder of the compound of Formula IV through two adjacent
carbons, wherein said C.sub.3--O.sub.6 carbocyclene is optionally
substituted with C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3
haloalkyl; L is C.sub.3-C.sub.6 alkylene, C.sub.3-C.sub.6
alkenylene or --(CH.sub.2).sub.3-cyclopropylene-, optionally
substituted with 1-4 halogen, --OH, or cyano; Q is C.sub.2-C.sub.4
alkyl or C.sub.3-C.sub.6 carbocyclyl optionally substituted with
C.sub.1-C.sub.3 alkyl, halogen, --OH, or cyano; E is
C.sub.1-C.sub.3 alkyl or C.sub.2-C.sub.3 alkenyl, optionally
substituted with 1-3 halogen; W is H, --OH,
--O(C.sub.1-C.sub.3)alkyl, --O(C.sub.1-C.sub.3)haloalkyl, halogen
or cyano; and Z.sup.2a is H or C.sub.1-C.sub.3 alkyl.
2. The compound of claim 1, or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof,
wherein J is C.sub.1-C.sub.3 alkyl.
3. The compound of claim 1, or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof,
wherein J is methyl or ethyl.
4. The compound of any one of claims 1 to 3, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein {circle around (T)} is C.sub.3-C.sub.6
carbocyclylene that is attached to L and to the remainder of the
compound of Formula IV through two adjacent carbons, wherein said
C.sub.3-C.sub.6 carbocyclene is optionally substituted with
C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3 haloalkyl.
5. The compound of any one of claims 1 to 3, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein {circle around (T)} is C.sub.3-C.sub.6
carbocyclylene that is attached to L and to the remainder of the
compound of Formula IV through two adjacent carbons, wherein the
C.sub.3-C.sub.6 carbocyclene is optionally substituted with methyl,
ethyl or trifluoromethyl.
6. The compound of any one of claims 1 to 3, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein {circle around (T)} is cyclopropylene.
7. The compound of any one of claims 1 to 3, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein {circle around (T)} is C.sub.6-C.sub.8 bridged
bicyclic carbocyclylene or C.sub.6-C.sub.8 fused bicyclic
carbocyclylene that is attached to L and to the remainder of the
compound of Formula IV through two adjacent carbons.
8. The compound of any one of claims 1 to 7, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein L is C.sub.3-C.sub.6 alkylene, substituted with
1-4 halogens.
9. The compound of any one of claims 1 to 7, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein L is C.sub.5 alkylene, substituted with two
halogens.
10. The compound of any one of claims 1 to 7, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein L is C.sub.3-C.sub.6 alkylene.
11. The compound of any one of claims 1 to 7, or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein L is C.sub.5 alkylene.
12. The compound of claim 8 or claim 9, or a stereoisomer, or a
mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, wherein the halogens are each fluoro.
13. The compound of any one of claims 1 to 12, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein Q is t-butyl or C.sub.5-C.sub.6
carbocyclyl.
14. The compound of any one of claims 1 to 12, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein Q is t-butyl.
15. The compound of any one of claims 1 to 14, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein E is C.sub.1-C.sub.3 alkyl optionally
substituted with 1-3 halogen atoms.
16. The compound of any one of claims 1 to 14, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein E is difluoromethyl.
17. The compound of any one of claims 1 to 16, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein W is hydrogen, --O(C.sub.1-C.sub.3)alkyl,
halogen or cyano.
18. The compound of any one of claims 1 to 16, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein W is methoxy.
19. The compound of any one of claims 1 to 18, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein Z.sup.2a is hydrogen or methyl.
20. The compound of any one of claims 1 to 18, or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, wherein Z.sup.2a is methyl.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 13/934,090, filed Jul. 2, 2013, which claims the benefit of
U.S. Provisional Application No. 61/798,524, filed on Mar. 15,
2013, and U.S. Application No. 61/667,806, filed Jul. 3, 2012;
which are herein incorporated by reference in their entirety.
FIELD
[0002] Novel small molecule inhibitors of viral replication are
disclosed, compositions containing such compounds, and therapeutic
methods comprising the administration of such compounds are also
disclosed.
BACKGROUND
[0003] The hepatitis C virus (HCV), a member of the hepacivirus
genera within the Flaviviridae family, is the leading cause of
chronic liver disease worldwide (Boyer, N. et al. J Hepatol. 2000,
32, 98-112). Consequently, a significant focus of current antiviral
research is directed toward the development of improved methods for
the treatment of chronic HCV infections in humans (Ciesek, S., von
Hahn T., and Manns, M P., Clin. Liver Dis., 2011, 15, 597-609;
Soriano, V. et al, J. Antimicrob. Chemother., 2011, 66, 1573-1686;
Brody, H., Nature Outlook, 2011, 474, S1-S7; Gordon, C. P., et al.,
J. Med. Chem. 2005, 48, 1-20; Maradpour, D., et al., Nat. Rev.
Micro. 2007, 5, 453-463).
[0004] Virologic cures of patients with chronic HCV infection are
difficult to achieve because of the prodigious amount of daily
virus production in chronically infected patients and the high
spontaneous mutability of HCV (Neumann, et al., Science 1998, 282,
103-7; Fukimoto, et al., Hepatology, 1996, 24, 1351-4; Domingo, et
al., Gene 1985, 40, 1-8; Martell, et al., J. Virol. 1992, 66,
3225-9). HCV treatment is further complicated by the fact that HCV
is genetically diverse and expressed as several different genotypes
and numerous subtypes. For example, HCV is currently classified
into six major genotypes (designated 1-6), many subtypes
(designated a, b, c, and so on), and about 100 different strains
(numbered 1, 2, 3, and so on).
[0005] HCV is distributed worldwide with genotypes 1, 2, and 3
predominate within the United States, Europe, Australia, and East
Asia (Japan, Taiwan, Thailand, and China). Genotype 4 is largely
found in the Middle East, Egypt and central Africa while genotype 5
and 6 are found predominantly in South Africa and South East Asia
respectively (Simmonds, P. et al. J Virol. 84: 4597-4610,
2010).
[0006] The combination of ribavirin, a nucleoside analog, and
interferon-alpha (a) (IFN), is utilized for the treatment of
multiple genotypes of chronic HCV infections in humans. However,
the variable clinical response observed within patients and the
toxicity of this regimen have limited its usefulness. Addition of a
HCV protease inhibitor (telaprevir or boceprevir) to the ribavirin
and IFN regimen improves 12-week post-treatment virological
response (SVR12) rates substantially. However, the regimen is
currently only approved for genotype 1 patients and toxicity and
other side effects remain.
[0007] The use of directing acting antivirals to treat multiple
genotypes of HCV infection has proven challenging due to the
variable activity of antivirals against the different gentoypes.
HCV protease inhibitors frequently have compromised in vitro
activity against HCV genotypes 2 and 3 compared to genotype 1 (See,
e.g., Table 1 of Summa, V. et al., Antimicrobial Agents and
Chemotherapy, 2012, 56, 4161-4167; Gottwein, J. et al,
Gastroenterology, 2011, 141, 1067-1079). Correspondingly, clinical
efficacy has also proven highly variable across HCV genotypes. For
example, therapies that are highly effective against HCV genotype 1
and 2 may have limited or no clinical efficacy against genotype 3.
(Moreno, C. et al., Poster 895, 61.sup.st AASLD Meeting, Boston,
Mass., USA, Oct. 29-Nov. 2, 2010; Graham, F., et al,
Gastroenterology, 2011, 141, 881-889; Foster, G. R. et al., EASL
45.sup.th Annual Meeting, Apr. 14-18, 2010, Vienna, Austria.) In
some cases, antiviral agents have good clinical efficacy against
genotype 1, but lower and more variable against genotypes 2 and 3.
(Reiser, M. et al., Hepatology, 2005, 41, 832-835.) To overcome the
reduced efficacy in genotype 3 patients, substantially higher doses
of antiviral agents may be required to achieve substantial viral
load reductions (Fraser, I P et al., Abstract #48, HEP DART 2011,
Koloa, Hi., December 2011.)
[0008] Antiviral agents that are less susceptible to viral
resistance are also needed. For example, resistance mutations at
positions 155 and 168 in the HCV protease frequently cause a
substantial decrease in antiviral efficacy of HCV protease
inhibitors (Mani, N. Ann Forum Collab HIV Res., 2012, 14, 1-8;
Romano, K P et al, PNAS, 2010, 107, 20986-20991; Lenz O,
Antimicrobial agents and chemotherapy, 2010, 54, 1878-1887.)
[0009] In view of the limitations of current HCV therapy, there is
a need to develop more effective anti-HCV therapies. It would also
be useful to provide therapies that are effective against multiple
HCV genotypes and subtypes.
SUMMARY
[0010] Novel compounds that inhibit the hepatitis C virus (HCV) NS3
protease are disclosed. In certain embodiments, the compounds
disclosed inhibit multiple genotypes of the hepatitis C virus.
These compounds are useful for the treatment of HCV infection and
the related symptoms.
[0011] In one embodiment, a compound of Formula (IV):
##STR00002##
or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof, is provided, wherein:
[0012] J is C.sub.1-C.sub.4 alkyl or C.sub.3-C.sub.6 carbocyclyl,
wherein C.sub.1-C.sub.4 alkyl or C.sub.3-C.sub.6 carbocyclyl is
optionally substituted with halogen, --OH, aryl or cyano; [0013]
{circle around (T)} is C.sub.3-C.sub.5 carbocyclylene that is
attached to L and to the remainder of the compound through two
adjacent carbons, wherein said C.sub.3-C.sub.6 carbocyclylene is
optionally substituted with C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.3
haloalkyl, halogen, --OH, or cyano, or {circle around (T)} is
C.sub.5-C.sub.8 bicyclic carbocyclylene that is attached to L and
to the remainder of the compound through two adjacent carbons;
[0014] L is C.sub.3-C.sub.6 alkylene, C.sub.3-C.sub.6 alkenylene or
--(CH.sub.2).sub.3-cyclopropyl-, optionally substituted with 1-4
halogen, --OH, or cyano; [0015] Q is C.sub.2-C.sub.4 alkyl or
C.sub.3-C.sub.6 carbocyclyl optionally substituted with
C.sub.1-C.sub.3 alkyl, halogen, --OH, or cyano; [0016] E is
C.sub.1-C.sub.3 alkyl or C.sub.2-C.sub.3 alkenyl, optionally
substituted with C.sub.1-C.sub.3 alkyl, halogen, --OH, or cyano;
[0017] W is H, --O(C.sub.1-C.sub.3)haloalkyl, halogen or cyano; and
[0018] Z.sup.2a is H or C.sub.1-C.sub.3 alkyl, halogen, --OH, or
cyano.
[0019] In a further embodiment of Formula (IV), {circle around (T)}
is C.sub.3-C.sub.6 carbocyclylene that is attached to L and to the
remainder of the compound of Formula IV through two adjacent
carbons, wherein said C.sub.3-C.sub.6 carbocyclene is optionally
substituted with C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3
haloalkyl.
[0020] In a further embodiment of Formula (IV), {circle around (T)}
is C.sub.3-C.sub.6 carbocyclylene that is attached to L and to the
remainder of the compound of Formula IV through two adjacent
carbons, wherein the C.sub.3-C.sub.6 carbocyclene is optionally
substituted with methyl, ethyl or trifluoromethyl.
[0021] In a further embodiment of Formula (IV), {circle around (T)}
is cyclopropylene.
[0022] In a further embodiment of Formula (IV), {circle around (T)}
is C.sub.6-C.sub.8 bridged bicyclic carbocyclylene or
C.sub.6-C.sub.8 fused bicyclic carbocyclylene that is attached to L
and to the remainder of the compound of Formula IV through two
adjacent carbons.
[0023] In a further embodiment of Formula (IV), L is
C.sub.3-C.sub.6 alkylene, substituted with 1-4 halogens. In another
embodiment of Formula (IV), L is C.sub.5 alkylene, substituted with
two halogens. In some embodiments, the halogens are each
fluoro.
[0024] In a further embodiment of Formula (IV), L is
C.sub.3-C.sub.6 alkylene.
[0025] In a further embodiment of Formula (IV), L is C.sub.5
alkylene.
[0026] In a further embodiment of Formula (IV), Q is t-butyl or
C.sub.5-C.sub.6 carbocyclyl.
[0027] In a further embodiment of Formula (IV), Q is t-butyl.
[0028] In a further embodiment of Formula (IV), E is
C.sub.1-C.sub.3 alkyl optionally substituted with 1-3 halogen
atoms.
[0029] In a further embodiment of Formula (IV), E is
difluoromethyl.
[0030] In a further embodiment of Formula (IV), W is hydrogen,
--O(C.sub.1-C.sub.3)alkyl, halogen or cyano.
[0031] In a further embodiment of Formula (IV), W is methoxy.
[0032] In a further embodiment of Formula (IV), Z.sup.2a is
hydrogen or methyl.
[0033] In a further embodiment of Formula (IV), Z.sup.2a is
methyl.
[0034] In one embodiment, a compound of Formula (I):
##STR00003##
or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof, is provided, wherein:
[0035] J is J.sup.1, J.sup.2, J.sup.3, J.sup.4, J.sup.5, J.sup.6,
J.sup.7, J.sup.8 or J.sup.9; [0036] {circle around (T)} is T.sup.1,
T.sup.2, T.sup.3, T.sup.4, T.sup.5, T.sup.6, T.sup.7, T.sup.8,
T.sup.9, T.sup.10, T.sup.11, T.sup.12, T.sup.13 or T.sup.14; [0037]
L is L.sup.1, L.sup.2, L.sup.3, L.sup.4, L.sup.5, L.sup.6, L.sup.7,
L.sup.8, L.sup.9 or L.sup.10; [0038] X is --O--, --CH.sub.2--,
--OC(O)--, --C(O)O--, --C(O)--, --SO.sub.2--, --S(O)--,
--N(R.sup.16)--, --S--, .dbd.N--O-- or a bond; [0039] A is
--C(O)--, --S(O).sub.2--, a 6-10 membered arylene, 5-10 membered
heteroarylene, or 4-10 membered heterocyclene, wherein any of said
arylene, heterocyclene, or heteroarylene is optionally substituted
with 1-4 Z.sup.1 groups; [0040] M is a bond, C.sub.1-C.sub.6
alkylene, --O--, or --N(R.sup.16)--; [0041] R.sup.1 is H or F;
[0042] R.sup.3, R.sup.4, and R.sup.5 are each independently
selected from H or Z.sup.1; [0043] Q is Q.sup.1, Q.sup.2, Q.sup.3,
Q.sup.4, Q.sup.5, Q.sup.6 or Q.sup.7; [0044] E is E.sup.1, E.sup.2,
E.sup.3, E.sup.4, E.sup.5, or E.sup.6; [0045] G is --CO.sub.2H,
--CONHSO.sub.2Z.sup.2, tetrazolyl, --CONHP(O)(R.sup.16).sub.2,
--P(O)(OH)(R.sup.16), and --P(O)(R.sup.16).sub.2; [0046] {circle
around (U)} is U.sup.1, U.sup.2, U.sup.3, U.sup.4, U.sup.5, U.sup.6
or U.sup.7; [0047] J.sup.1 is halogen; [0048] J.sup.2 is --OH and
R.sup.1 is H; [0049] J.sup.3 is --NR.sup.17R.sup.18 and R.sup.1 is
H; [0050] J.sup.4 is C.sub.1-C.sub.8 alkyl; [0051] J.sup.5 is
C.sub.1-C.sub.8 alkyl substituted with 1-4 Z.sup.3 groups; [0052]
J.sup.6 is C.sub.3-C.sub.8 carbocyclyl optionally substituted with
1-4 Z.sup.3 groups; [0053] J.sup.7 is C.sub.6-C.sub.10 aryl, 5-10
membered heteroaryl, or 4-10 membered heterocyclyl optionally
substituted with 1-4 Z.sup.3 groups; [0054] J.sup.8 is
C.sub.1-C.sub.8 alkoxy optionally substituted with 1-4 Z.sup.3
groups and R.sup.1 is H; [0055] J.sup.9 is C.sub.3-C.sub.8
carbocyclyleoxy optionally substituted with 1-4 Z.sup.3 groups and
R.sup.1 is H; [0056] T.sup.1 is C.sub.3-C.sub.8 carbocyclylene that
is attached to L and M through two adjacent carbons; [0057] T.sup.2
is C.sub.3-C.sub.8 carbocyclylene that is attached to L and M
through two adjacent carbons, wherein said carbocyclylene is
substituted with 1-4 C.sub.1-C.sub.8 alkyl groups; [0058] T.sup.3
is C.sub.3-C.sub.8 carbocyclylene that is attached to L and M
through two adjacent carbons, wherein said carbocyclylene is
substituted with 1-4 halogen atoms and said carbocyclylene is
optionally substituted with 1-4 C.sub.1-C.sub.6 alkyl groups;
[0059] T.sup.4 is C.sub.3-C.sub.8 carbocyclylene that is attached
to L and M through two adjacent carbons, wherein said
carbocyclylene is optionally substituted with a C.sub.1-C.sub.8
alkyl group, wherein said alkyl group is optionally substituted
with 1-4 Z.sup.3 groups; [0060] T.sup.5 is 4-10 membered
heterocyclene that is attached to L and M through two adjacent
carbons; [0061] T.sup.6 is 4-10 membered heterocyclene that is
attached to L through a carbon atom and attached to M through an N
atom, wherein said heterocyclene is optionally substituted with 1-4
Z.sup.1 groups; [0062] T.sup.7 is 4-10 membered heterocyclene that
is attached to M through a carbon atom and attached to L through an
N atom, wherein said heterocyclene is optionally substituted with
1-4 Z.sup.1 groups; [0063] T.sup.8 is 4-10 membered heterocyclene
that is attached to L and M through two adjacent carbons, wherein
said heterocyclene is optionally substituted with 1-4 Z.sup.1
groups; [0064] T.sup.9 is C.sub.5-C.sub.12 spiro bicyclic
carbocyclylene that is attached to L and M through two adjacent
carbons, wherein said spiro bicyclic carbocyclylene is optionally
substituted with 1-4 Z.sup.1 groups; [0065] T.sup.10 is
C.sub.5-C.sub.12 fused bicyclic carbocyclylene that is attached to
L and M through two adjacent carbons, wherein said fused bicyclic
carbocyclylene is optionally substituted with 1-4 Z.sup.1 groups;
[0066] T.sup.11 is C.sub.5-C.sub.12 bridged bicyclic carbocyclylene
that is attached to L and M through two adjacent carbons, wherein
said bridged bicyclic carbocyclylene is optionally substituted with
1-4 Z.sup.1 groups; [0067] T.sup.12 is C.sub.4-C.sub.8
carbocyclylene that is attached to L and M through two non-adjacent
carbons, wherein said carbocyclylene is optionally substituted with
1-4 Z.sup.1 groups; [0068] T.sup.13 is a 5-8 membered fused,
bridged, or spiro bicyclic heterocyclene that is attached to L and
M through two adjacent atoms, wherein said heterocyclene is
optionally substituted with 1-4 Z.sup.1 groups; [0069] T.sup.14 is
C.sub.3-C.sub.8 carbocyclylene that is attached to L and M through
two adjacent carbons, wherein said carbocyclylene is optionally
substituted with 1-4 Z.sup.4 groups; [0070] L.sup.1 is
C.sub.1-C.sub.8 alkylene or C.sub.2-C.sub.8 alkenylene; [0071]
L.sup.2 is C.sub.1-C.sub.8 alkylene or C.sub.2-C.sub.8 alkenylene
wherein said C.sub.1-C.sub.8 alkylene is substituted with 1-4
halogens or said C.sub.2-C.sub.8 alkenylene is substituted with 1-4
halogens; [0072] L.sup.3 is C.sub.1-C.sub.8 alkylene or
C.sub.2-C.sub.8 alkenylene wherein said C.sub.1-C.sub.8 alkylene is
substituted with 1-4 Z.sup.4 groups or said C.sub.2-C.sub.8
alkenylene is substituted with 1-4 Z.sup.4 groups and wherein each
is optionally substituted with 1-4 halogens; [0073] L.sup.4 is
C.sub.1-C.sub.8 alkylene or C.sub.2-C.sub.8 alkenylene substituted
with two geminal C.sub.1-C.sub.4 alkyl groups that come together to
form a spiro C.sub.3-C.sub.8 carbocyclyl group, wherein L.sup.4 is
optionally substituted with 1-4 Z.sup.1 groups; [0074] L.sup.5 is
2-8 membered heteroalkylene or 4-8 membered heteroalkenylene that
is connected to {circle around (T)} by an O, S or N atom and said
heteroalkylene or heteroalkenylene is optionally substituted with
1-4 Z.sup.3 groups; [0075] L.sup.6 is 2-8 membered heteroalkylene
or 5-8 membered heteroalkenylene that is connected to {circle
around (T)} by a carbon atom and said heteroalkylene or
heteroalkenylene is substituted with 1-4 halogen atoms and is
optionally substituted with 1-4 Z.sup.4 groups; [0076] L.sup.7 is
2-8 membered heteroalkylene or 4-8 membered heteroalkenylene that
is connected to {circle around (T)} by a carbon atom and said
heteroalkylene or heteroalkenylene is optionally substituted with
1-4 Z.sup.4 groups; [0077] L.sup.8 is L.sup.8A-L.sup.8B-L.sup.8C
wherein L.sup.8A and L.sup.8C are each independently selected from
C.sub.1-C.sub.6 alkylene, C.sub.1-C.sub.6 heteroalkylene,
C.sub.2-C.sub.6 alkenylene or a bond and L.sup.8B is a 3- to
6-membered saturated or unsaturated ring containing 0 to 3
heteroatoms selected from N, O, or S, wherein L.sup.8A and L.sup.8C
connect to L.sup.8B at two different ring atoms and L.sup.8B is
optionally substituted with 1-4 Z.sup.1 groups; [0078] L.sup.9 is
C.sub.2-C.sub.8 alkynylene optionally substituted with 1-4 Z.sup.1
groups; [0079] L.sup.10 is C.sub.1-C.sub.8 alkylene or
C.sub.3-C.sub.8 alkenylene substituted with two geminal Z.sup.1
groups that come together to form a spiro 4-8 membered heterocyclyl
group, wherein L.sup.10 is optionally substituted with 1-4 Z.sup.1
groups; [0080] U.sup.1 is C.sub.6-C.sub.14 membered arylene
optionally substituted with 1-4 W groups; [0081] U.sup.2 is
C.sub.3-C.sub.8 membered carbocyclylene optionally substituted with
1-4 W groups; [0082] U.sup.3 is 4-14 membered heterocyclene
optionally substituted with 1-4 W groups that are located on one or
more ring atoms selected from C or N; [0083] U.sup.4 is 5 or 6
membered monocyclic heteroarylene containing 1, 2 or 3 heteroatoms
independently selected from N, O, or S, wherein said heteroarylene
is optionally substituted with 1-4 W groups that are located on one
or more ring atoms selected from C or N; [0084] U.sup.5 is 8, 9 or
10 membered fused bicyclic heteroarylene containing 1, 2 or 3
heteroatom ring atoms independently selected from N, O, or S,
wherein said heteroarylene is optionally substituted with 1-4 W
groups that are located on one or more ring atoms selected from C
or N; [0085] U.sup.6 is 11-14 membered fused tricyclic
heteroarylene containing 1, 2, 3 or 4 heteroatom ring atoms
independently selected from N, O, or S, wherein said heteroarylene
is optionally substituted with 1-4 W groups that are located on one
or more ring atoms selected from C or N; [0086] U.sup.7 is 8-10
membered fused bicyclic heteroarylene containing 4 heteroatom ring
atoms independently selected from N, O, or S, wherein said
heteroaryl is optionally substituted with 1-2 W groups that are
located on one or more ring atoms selected from C or N; [0087] W is
independently W.sup.1, W.sup.2, W.sup.3, W.sup.4, W.sup.5, W.sup.6
or W.sup.7; [0088] W.sup.1 is oxo, halogen, --OR.sup.6,
C.sub.1-C.sub.6 alkyl, --CN, --CF.sub.3, --SR.sup.6,
--C(O).sub.2R.sup.6, --C(O)N(R.sup.6).sub.2, --C(O)R.sup.6,
--N(R.sup.6)C(O)R.sup.6, --SO.sub.2(C.sub.1-C.sub.6 alkyl),
--S(O)(C.sub.1-C.sub.6 alkyl), C.sub.3-C.sub.8 carbocyclyl,
C.sub.3-C.sub.8 cycloalkoxy, C.sub.1-C.sub.6 haloalkyl,
--N(R.sup.6).sub.2, --NR.sup.6(C.sub.1-C.sub.6
alkyl)O(C.sub.1-C.sub.6 alkyl), halo(C.sub.1-C.sub.6 alkoxy),
--NR.sup.6SO.sub.2R.sup.6, --SO.sub.2N(R.sup.6).sub.2,
--NHCOOR.sup.6, --NHCONHR.sup.6, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl or --O(4-10
membered heterocyclyl), wherein said W.sup.1 alkyl, carbocyclyl,
cycloalkoxy, haloalkyl, haloalkoxy, aryl, heteroaryl, or
heterocyclyl is optionally substituted with 1-4 Z.sup.1c groups;
[0089] each R.sup.6 is independently selected from H,
C.sub.6-C.sub.10 aryl or C.sub.1-C.sub.6 alkyl, wherein said aryl
or alkyl is optionally substituted with 1 to 4 substituents
independently selected from halogen atoms, C.sub.1-C.sub.6 alkyl,
C.sub.6-C.sub.10 aryl, C.sub.3-C.sub.8 carbocyclyl, 5-14 membered
heteroaryl, 4-10 membered heterocyclyl, halo(C.sub.1-C.sub.6
alkoxy), --OH, --O(C.sub.1-C.sub.6 alkyl), --SH,
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --C(O)(C.sub.1-C.sub.6
alkyl), --SO.sub.2N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHCOO(C.sub.1-C.sub.6 alkyl), --NHCO(C.sub.1-C.sub.6 alkyl),
--NHCONH(C.sub.1-C.sub.6 alkyl), --CO.sub.2(C.sub.1-C.sub.6 alkyl),
or --C(O)N(C.sub.1-C.sub.6 alkyl).sub.2; [0090] W.sup.2 is
C.sub.1-C.sub.6 alkoxy substituted with a 5-14 membered heteroaryl
or C.sub.6-C.sub.10 aryl; wherein said heteroaryl or aryl is
substituted with 1-4 Z.sup.1 groups; [0091] W.sup.3 is
C.sub.2-C.sub.6 alkynyl substituted with an C.sub.6-C.sub.10 aryl,
C.sub.3-C.sub.8 carbocyclyl, C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.6
haloalkyl, 4-10 membered heterocyclyl, or 5-14 membered heteroaryl;
wherein said aryl, carbocyclyl, alkyl, haloalkyl, heterocyclyl, or
heteroaryl is optionally substituted with 1-4 Z.sup.1 groups;
[0092] W.sup.4 is --SF.sub.5; [0093] W.sup.5 is --O(C.sub.2-C.sub.6
alkyl)OR.sup.22 wherein R.sup.22 is an C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl or 4-10 membered heterocyclyl optionally
substituted with 1-4 Z.sup.1 groups; [0094] W.sup.6 is
--O(C.sub.2-C.sub.6 alkyl)NR.sup.16R.sup.22 wherein R.sup.22 is an
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or 4-10 membered
heterocyclyl optionally substituted with 1-4 Z.sup.1 groups; [0095]
W.sup.7 is --O(5-14 membered heteroaryl); wherein said --O(5-14
membered heteroaryl) is optionally substituted with 1-4 Z.sup.1
groups; [0096] E.sup.1 is C.sub.2-C.sub.6 alkenyl; [0097] E.sup.2
is C.sub.1-C.sub.6 alkyl; [0098] E.sup.3 is C.sub.1-C.sub.6
haloalkyl; [0099] E.sup.4 is C.sub.2-C.sub.6 haloalkenyl; [0100]
E.sup.5 is C.sub.3-C.sub.6 carbocyclyl; [0101] E.sup.6 is
C.sub.1-C.sub.6 alkyl substituted with --OCH.sub.3, --OCD.sub.3,
--OCF.sub.3, or --OCF.sub.2H; [0102] Q.sup.1 is H, C.sub.1-C.sub.8
alkyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.6-C.sub.10 aryl, 5-6
membered heteroaryl, or 5-6 membered heterocyclyl, wherein when
Q.sup.1 is not H, said Q.sup.1 is optionally substituted with 1-3
substituents independently selected from halogen, --OR.sup.6,
--SR.sup.6, --N(R.sup.6).sub.2, C.sub.6-C.sub.10 aryl,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, --CN, --CF.sub.3, --SO.sub.2(C.sub.1-C.sub.6 alkyl),
--S(O)(C.sub.1-C.sub.6 alkyl), --NR.sup.6SO.sub.2Z.sup.2,
--SO.sub.2NR.sup.17R.sup.18, --NHCOOR.sup.16, --NHCOZ.sup.2,
--NHCONHR.sup.16, --CO.sub.2R.sup.6, --C(O)R.sup.6, or
--CON(R.sup.6).sub.2; [0103] Q.sup.2 is C.sub.5-C.sub.10 spiro
bicyclic carbocyclyl optionally substituted with 1-4 Z.sup.1
groups; [0104] Q.sup.3 is C.sub.5-C.sub.10 fused bicyclic
carbocyclyl optionally substituted with 1-4 Z.sup.1 groups; [0105]
Q.sup.4 is C.sub.5-C.sub.10 bridged bicyclic carbocyclyl optionally
substituted with 1-4 Z.sup.1 groups; [0106] Q.sup.5 is 4-membered
heterocyclyl having 1 heteroatom selected from N, O or S wherein
Q.sup.5 is optionally substituted with 1-4 Z.sup.3 groups; [0107]
Q.sup.6 is C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.6-C.sub.10 aryl, 5-6 membered heteroaryl, or 5-6 membered
heterocyclyl, wherein Q.sup.6 is substituted with 1 oxo group and
with 0 to 3 substituents independently selected from halogen,
--OR.sup.6, --SR.sup.6, --N(R.sup.6).sub.2, C.sub.6-C.sub.10 aryl,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, --NO.sub.2, --CN, --CF.sub.3,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --S(O)(C.sub.1-C.sub.6 alkyl),
--NR.sup.6SO.sub.2Z.sup.2, --SO.sub.2NR.sup.17R.sup.18,
--NHCOOR.sup.16, --NHCOZ.sup.2, --NHCONHR.sup.16,
--CO.sub.2R.sup.6, --C(O)R.sup.6, or --CON(R.sup.6).sub.2; [0108]
Q.sup.7 is C.sub.3-C.sub.8 carbocyclyl, wherein Q.sup.7 is
substituted with 4-8 F atoms and each carbon of Q.sup.7 is
substituted with 0-2 F atoms; [0109] each Z.sup.1 is independently
oxo, halogen, C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8 alkenyl,
C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8 haloalkyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10 membered
heterocyclyl, --CN, --C(O)R.sup.16, --C(O)OR.sup.16,
--C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)R.sup.16, --NR.sup.16C(O)NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2R.sup.16,
--NR.sup.16S(O).sub.2NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2OR.sup.16, --OR.sup.16, --OC(O)R.sup.16,
--OC(O)NR.sup.17R.sup.18, --SR.sup.16, --S(O)R.sup.16,
--S(O).sub.2R.sup.16 or --S(O).sub.2NR.sup.17R.sup.18 wherein any
alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl or
heterocyclyl of Z.sup.1 is optionally substituted with 1-4 Z.sup.1a
groups; [0110] each Z.sup.1a is independently oxo, halogen,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8
haloalkyl, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10
membered heterocyclyl, --CN, --C(O)R.sup.16, --C(O)OR.sup.16
, --C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)R.sup.16, --NR.sup.16C(O)OR.sup.16,
--NR.sup.16C(O)NR.sup.17R.sup.18, --NR.sup.16S(O).sub.2R.sup.16,
--NR.sup.16S(O).sub.2NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2OR.sup.16, --OR.sup.16, --OC(O)R.sup.16,
--OC(O)NR.sup.17R.sup.18, --SR.sup.16, --S(O)R.sup.16,
--S(O).sub.2R.sup.16 or --S(O).sub.2NR.sup.17R.sup.18 wherein any
alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclyl of
Z.sup.1a is optionally substituted with 1-4 Z.sup.1c groups; [0111]
each R.sup.16 is independently H, C.sub.1-C.sub.8 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or 4-10 membered
heterocyclyl, wherein any alkyl, alkenyl, alkynyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl of R.sup.16 is optionally
substituted with 1-4 Z.sup.1c groups; [0112] each Z.sup.1c is
independently oxo, halogen, C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8
haloalkyl, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10
membered heterocyclyl, --CN, --C(O)(C.sub.1--C.sub.8 alkyl),
--C(O)O(C.sub.1-C.sub.8 alkyl), --C(O)N(C.sub.1-C.sub.8
alkyl).sub.2, --NH.sub.2, --NH(C.sub.1-C.sub.8 alkyl),
--N(C.sub.1-C.sub.8 alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.8 alkyl),
--NHC(O)(C.sub.1-C.sub.8 alkyl), --NHC(O)NH(C.sub.1-C.sub.8 alkyl),
--OH, --O(C.sub.1-C.sub.8 alkyl), C.sub.3-C.sub.8 cycloalkoxy,
C.sub.5-C.sub.10 bicyclic carbocyclyloxy, --S(C.sub.1-C.sub.8
alkyl) or --S(O).sub.2N(C.sub.1-C.sub.8 alkyl).sub.2 wherein any
alkyl, carbocyclyl, aryl, heteroaryl, heterocyclyl or cycloalkoxy
portion of Z.sup.1c is optionally substituted with 1-4 halogen
atoms or C.sub.1-C.sub.6 alkoxy groups; [0113] R.sup.17 and
R.sup.18 are each independently H, C.sub.1-C.sub.8 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, --C(O)R.sup.16,
--C(O)OR.sup.16, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or
4-10 membered heterocyclyl, wherein any alkyl, alkenyl, alkynyl,
carbocyclyl, aryl, heteroaryl or heterocyclyl of R.sup.17 or
R.sup.18 is optionally substituted with 1-4 Z.sup.1c groups, or
R.sup.17 and R.sup.18 together with the nitrogen to which they are
attached form a 4-7 membered heterocyclyl group, wherein said 4-7
membered heterocyclyl group is optionally substituted with 1-4
Z.sup.1c groups; [0114] each Z.sup.2 is independently
C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl,
--NR.sup.17R.sup.18 or --OR.sup.16 wherein any alkyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl portion of Z.sup.2 is optionally
substituted with 1-4 Z.sup.2a groups; [0115] each Z.sup.2a is
independently oxo, halogen, C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.5-C.sub.10 bicyclic
carbocyclyl, C.sub.1-C.sub.8 haloalkyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl, --(C.sub.2-C.sub.8
alkynyl)aryl, --(C.sub.2-C.sub.8 alkynyl)heteroaryl, --CN,
--C(O)(C.sub.1-C.sub.6 alkyl), --C(O)O(C.sub.1-C.sub.6 alkyl),
--C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)O(C.sub.1-C.sub.6 alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --OH, --O(C.sub.1-C.sub.6
alkyl), halo(C.sub.1-C.sub.6 alkoxy), C.sub.3-C.sub.8 cycloalkoxy,
--S(C.sub.1-C.sub.6 alkyl), or --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2; wherein any alkyl, alkynyl, carbocyclyl, cycloalkoxy,
aryl, heteroaryl or heterocyclyl portions of Z.sup.2a is optionally
substituted with 1-4 halogen or C.sub.1-C.sub.6 alkoxy groups;
[0116] each Z.sup.3 is independently oxo, halogen, C.sub.2-C.sub.8
alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8 haloalkyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10 membered
heterocyclyl, --CN, --C(O)OR.sup.16, --C(O)NR.sup.17R.sup.18,
--NR.sup.17R.sup.18, --NR.sup.16C(O)NR.sup.17R.sup.18, --OR.sup.16,
--SR.sup.16 or --SO.sub.2R.sup.16; wherein any alkenyl, alkynyl,
carbocyclyl, aryl, heteroaryl or heterocyclyl portions of Z.sup.3
is optionally substituted with 1-4 halogen; and [0117] each Z.sup.4
is independently oxo, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.5-C.sub.10 bicyclic
carbocyclyl, C.sub.1-C.sub.8 haloalkyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl, --CN,
--C(O)OR.sup.16, --C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)NR.sup.17R.sup.18, --OR.sup.16, --SR.sup.16 or
--SO.sub.2R.sup.16, wherein any alkenyl, alkynyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl portions of Z.sup.4 is optionally
substituted with 1-4 halogen.
[0118] In another embodiment, a compound of Formula (II):
##STR00004##
or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof, is provided, wherein:
[0119] M is --O--; [0120] J is J.sup.1, J.sup.2, J.sup.3, J.sup.4,
J.sup.5, J.sup.6, J.sup.7, J.sup.8 or J.sup.9; [0121] {circle
around (T)} is T.sup.1, T.sup.2, T.sup.3, T.sup.4, T.sup.5,
T.sup.7, T.sup.8, T.sup.9, T.sup.1, T.sup.11, T.sup.12, T.sup.13 or
T.sup.14; [0122] L is L.sup.1, L.sup.2, L.sup.3, L.sup.4, L.sup.5,
L.sup.6, L.sup.7, L.sup.8, L.sup.9 or L.sup.10; [0123] R.sup.1 is H
or F; [0124] Q is Q.sup.1, Q.sup.2, Q.sup.3, Q.sup.4, Q.sup.5,
Q.sup.6 or Q.sup.7; [0125] E is E.sup.1, E.sup.2, E.sup.3, E.sup.4,
E.sup.5, or E6; [0126] {circle around (U)} is U.sup.1, U.sup.2,
U.sup.3, U.sup.4, U.sup.5, U.sup.6 or U.sup.7; [0127] J.sup.1 is
halogen; [0128] J.sup.2 is --OH and R.sup.1 is H; [0129] J.sup.3 is
--NR.sup.17R.sup.18 and R.sup.1 is H; [0130] J.sup.4 is
C.sub.1-C.sub.8 alkyl; [0131] J.sup.5 is C.sub.1-C.sub.8 alkyl
optionally substituted with 1-4 Z.sup.3 groups; [0132] J.sup.6 is
C.sub.3-C.sub.8 carbocyclyl optionally substituted with 1-4 Z.sup.3
groups; [0133] J.sup.7 is C.sub.6-C.sub.10 aryl, 5-14 membered
heteroaryl, or 4-10 membered heterocyclyl, any of which are
optionally substituted with 1-4 Z.sup.3 groups; [0134] J.sup.8 is
C.sub.1-C.sub.8 alkoxy optionally substituted with 1-4 Z.sup.3
groups and R.sup.1 is H; [0135] J.sup.9 is C.sub.3-C.sub.8
carbocyclyloxy optionally substituted with 1-4 Z.sup.3 groups and
R.sup.1 is H; [0136] T.sup.1 is C.sub.3-C.sub.8 carbocyclylene that
is attached to L and M through two adjacent carbons; [0137] T.sup.2
is C.sub.3-C.sub.8 carbocyclylene that is attached to L and M
through two adjacent carbons, wherein said carbocyclylene is
optionally substituted with 1-4 C.sub.1-C.sub.8 alkyl groups;
[0138] T.sup.3 is C.sub.3-C.sub.8 carbocyclylene that is attached
to L and M through two adjacent carbons, wherein said
carbocyclylene is optionally substituted with 1-4 halogen atoms and
said carbocyclylene is optionally substituted with 1-4
C.sub.1-C.sub.6 alkyl groups; [0139] T.sup.4 is C.sub.3-C.sub.8
carbocyclylene that is attached to L and M through two adjacent
carbons, wherein said carbocyclylene is optionally substituted with
a C.sub.1-C.sub.8 alkyl group, wherein said alkyl group is
optionally substituted with 1-4 Z.sup.3 groups; [0140] T.sup.5 is
4-10 membered heterocyclene that is attached to L and M through two
adjacent carbons; [0141] T.sup.7 is 4-10 membered heterocyclene
that is attached to M through a carbon atom and attached to L
through a N atom, wherein said heterocyclene is optionally
substituted with 1-4 Z.sup.1 groups; [0142] T.sup.8 is 4-10
membered heterocyclene that is attached to L and M through two
adjacent carbons, wherein said heterocyclene is optionally
substituted with 1-4 Z.sup.1 groups; [0143] T.sup.9 is
C.sub.5-C.sub.12 spiro bicyclic carbocyclylene that is attached to
L and M through two adjacent carbons, wherein said spiro bicyclic
carbocyclylene is optionally substituted with 1-4 Z.sup.1 groups;
[0144] T.sup.10 is C.sub.5-C.sub.12 fused bicyclic carbocyclylene
that is attached to L and M through two adjacent carbons, wherein
said fused bicyclic carbocyclylene is optionally substituted with
1-4 Z.sup.1 groups; [0145] T.sup.11 is C.sub.5-C.sub.12 bridged
bicyclic carbocyclylene that is attached to L and M through two
adjacent carbons, wherein said bridged bicyclic carbocyclylene is
optionally substituted with 1-4 Z.sup.1 groups; [0146] T.sup.12 is
C.sub.4-C.sub.8 carbocyclylene that is attached to L and M through
two non-adjacent carbons, wherein said carbocyclylene is optionally
substituted with 1-4 Z.sup.1 groups; [0147] T.sup.13 is a 5-8
membered fused, bridged, or spiro bicyclic heterocyclene that is
attached to L and M through two adjacent atoms, wherein said
heterocyclene is optionally substituted with 1-4 Z.sup.1 groups;
[0148] T.sup.14 is C.sub.3-C.sub.8 carbocyclylene that is attached
to L and M through two adjacent carbons, wherein said
carbocyclylene is optionally substituted with 1-4 Z.sup.4 groups;
[0149] L.sup.1 is C.sub.1-C.sub.8 alkylene or C.sub.2-C.sub.8
alkenylene; [0150] L.sup.2 is C.sub.1-C.sub.8 alkylene or
C.sub.2-C.sub.8 alkenylene wherein said C.sub.1-C.sub.8 alkylene is
substituted with 1-4 halogens or said C.sub.2-C.sub.8 alkenylene is
substituted with 1-4 halogens; [0151] L.sup.3 is C.sub.1-C.sub.8
alkylene or C.sub.2-C.sub.8 alkenylene wherein said C.sub.1-C.sub.8
alkylene is substituted with 1-4 Z.sup.4 groups or said
C.sub.2-C.sub.8 alkenylene is substituted with 1-4 Z.sup.4 groups
and wherein each is optionally substituted with 1-4 halogens;
[0152] L.sup.4 is C.sub.1-C.sub.8 alkylene or C.sub.2-C.sub.8
alkenylene substituted with two geminal C.sub.1-C.sub.4 alkyl
groups that come together to form a spiro C.sub.3-C.sub.8
carbocyclyl group, wherein L.sup.4 is optionally substituted with
1-4 Z.sup.1 groups; [0153] L.sup.5 is 2-8 membered heteroalkylene
or 4-8 membered heteroalkenylene that is connected to {circle
around (T)} by an O, S or N atom and said heteroalkylene or
heteroalkenylene is optionally substituted with 1-4 Z.sup.3 groups;
[0154] L.sup.6 is 2-8 membered heteroalkylene or 5-8 membered
heteroalkenylene that is connected to {circle around (T)} by a
carbon atom and said heteroalkylene or heteroalkenylene is
substituted with 1-4 halogen atoms and is optionally substituted
with 1-4 Z.sup.4 groups; [0155] L.sup.7 is 2-8 membered
heteroalkylene or 4-8 membered heteroalkenylene that is connected
to {circle around (T)} by a carbon atom and said heteroalkylene or
heteroalkenylene is optionally substituted with 1-4 Z.sup.4 groups;
[0156] L.sup.8 is L.sup.8A-L.sup.8B-L.sup.8 wherein L.sup.8A and
L.sup.8C are each independently selected from C.sub.1-C.sub.6
alkylene, C.sub.1-C.sub.6 heteroalkylene, C.sub.2-C.sub.6
alkenylene or a bond and L.sup.8B is a 3- to 6-membered saturated
or unsaturated ring containing 0 to 3 heteroatoms selected from N,
O, or S, wherein L.sup.8A and L.sup.8C connect to L.sup.8B at two
different ring atoms and L.sup.8B is optionally substituted with
1-4 Z.sup.1 groups; [0157] L.sup.9 is C.sub.2-C.sub.8 alkynylene
optionally substituted with 1-4 Z.sup.1 groups; [0158] L.sup.10 is
C.sub.1-C.sub.8 alkylene or C.sub.3-C.sub.8 alkenylene substituted
with two geminal Z.sup.1 groups that come together to form a spiro
4-8 membered heterocyclyl group, wherein L.sup.10 is optionally
substituted with 1-4 Z.sup.1 groups; [0159] U.sup.1 is
C.sub.6-C.sub.14 membered arylene optionally substituted with 1-4 W
groups; [0160] each U.sup.2 is C.sub.3-C.sub.8 membered
carbocyclylene optionally substituted 1-4 W groups; [0161] each
U.sup.3 is 4-14 membered heterocyclene optionally substituted with
1-4 W groups that are located on one or more ring atoms selected
from C or N; [0162] U.sup.4 is 5 or 6 membered monocyclic
heteroarylene containing 1, 2 or 3 heteroatoms independently
selected from N, O, or S, wherein said heteroarylene is optionally
substituted with 1-4 W groups that are located on one or more ring
atoms selected from C or N; [0163] U.sup.5 is 8, 9 or 10 membered
fused bicyclic heteroarylene containing 1, 2 or 3 heteroatom ring
atoms independently selected from N, O, or S, wherein said
heteroarylene is optionally substituted with 1-4 W groups that are
located on one or more ring atoms selected from C or N; [0164]
U.sup.6 is 11-14 membered fused tricyclic heteroarylene containing
1, 2, 3 or 4 heteroatom ring atoms independently selected from N,
O, or S, wherein said heteroarylene is optionally substituted with
1-4 W groups that are located on one or more ring atoms selected
from C or N; [0165] U.sup.7 is 8-10 membered fused bicyclic
heteroarylene containing 4 heteroatom ring atoms independently
selected from N, O, or S, wherein said heteroaryl is optionally
substituted with 1-2 W groups that are located on one or more ring
atoms selected from C or N; [0166] each W is independently W.sup.1,
W.sup.2, W.sup.3, W.sup.4, W.sup.5, W.sup.6 or W.sup.7; [0167] each
W.sup.1 is oxo, halogen, --OR.sup.6, C.sub.1-C.sub.6 alkyl, --CN,
--CF.sub.3, --SR.sup.6, --C(O).sub.2R.sup.6,
--C(O)N(R.sup.6).sub.2, --C(O)R.sup.6, --N(R.sup.6)C(O)R.sup.6,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --S(O)(C.sub.1-C.sub.6 alkyl),
C.sub.3-C.sub.8 carbocyclyl, C.sub.3-C.sub.8 cycloalkoxy,
C.sub.1-C.sub.6 haloalkyl, --N(R.sup.6).sub.2,
--NR.sup.6(C.sub.1-C.sub.6 alkyl)O(C.sub.1-C.sub.6 alkyl),
halo(C.sub.1-C.sub.6 alkoxy), --NR.sup.6SO.sub.2R.sup.6,
--SO.sub.2N(R.sup.6).sub.2, --NHCOOR.sup.6, --NHCONHR.sup.6,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10 membered
heterocyclyl or --O(4-10 membered heterocyclyl), wherein said
W.sup.1 alkyl, carbocyclyl, cycloalkoxy, haloalkyl, haloalkoxy,
aryl, heteroaryl, or heterocyclyl is optionally substituted with
1-4 Z.sup.1c groups; [0168] each R.sup.6 is independently selected
from H, C.sub.6-C.sub.10 aryl or C.sub.1-C.sub.6 alkyl, wherein
said aryl or alkyl is optionally substituted with 1 to 4
substituents independently selected from halogen atoms,
C.sub.1-C.sub.6 alkyl, C.sub.6-C.sub.10 aryl, C.sub.3-C.sub.8
carbocyclyl, 5-14 membered heteroaryl, 4-10 membered heterocyclyl,
halo(C.sub.1-C.sub.6 alkoxy), --OH, --O(C.sub.1-C.sub.6 alkyl),
--SH, --S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --C(O)(C.sub.1-C.sub.6
alkyl), --SO.sub.2N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHCOO(C.sub.1-C.sub.6 alkyl), --NHCO(C.sub.1-C.sub.6 alkyl),
--NHCONH(C.sub.1-C.sub.6 alkyl), --CO.sub.2(C.sub.1-C.sub.6 alkyl),
or --C(O)N(C.sub.1-C.sub.6 alkyl).sub.2; [0169] each W.sup.2 is
C.sub.1-C.sub.6 alkoxy substituted with a 5-14 membered heteroaryl
or C.sub.6-C.sub.10 aryl; wherein said heteroaryl or aryl is
substituted with 1-4 Z.sup.1C groups; [0170] each W.sup.3 is
C.sub.2-C.sub.6 alkynyl substituted with an C.sub.6-C.sub.10 aryl,
C.sub.3-C.sub.8 carbocyclyl, C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.6
haloalkyl, 4-10 membered heterocyclyl, or 5-14 membered heteroaryl;
wherein said aryl, carbocyclyl, alkyl, haloalkyl, heterocyclyl, or
heteroaryl is optionally substituted with 1-4 Z.sup.1 groups;
[0171] each W.sup.4 is --SF.sub.5; [0172] each W.sup.5 is
--O(C.sub.2-C.sub.6 alkyl)OR.sup.22 wherein R.sup.22 is an
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or 4-10 membered
heterocyclyl optionally substituted with 1-4 Z.sup.1 groups; [0173]
each W.sup.6 is --O(C.sub.2-C.sub.6 alkyl)NR.sup.16R.sup.22 wherein
R.sup.22 is an C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or
4-10 membered heterocyclyl optionally substituted with 1-4 Z.sup.1
groups; [0174] each W.sup.7 is --O(5-14 membered heteroaryl);
wherein said --O(5-14 membered heteroaryl) is optionally
substituted with 1-4 Z.sup.1 groups and 2 adjacent substituents of
said --O(5-14 membered heteroaryl) may be taken together to form a
3- to 6-membered cyclic ring containing 0 to 3 heteroatoms
independently selected from N, O, or S; [0175] E.sup.1 is
C.sub.2-C.sub.6 alkenyl; [0176] E.sup.2 is O.sub.1--C.sub.6 alkyl;
[0177] E.sup.3 is C.sub.1-C.sub.6 haloalkyl; [0178] E.sup.4 is
C.sub.2-C.sub.6 haloalkenyl; [0179] E.sup.5 is C.sub.3-C.sub.6
carbocyclyl; [0180] E.sup.6 is C.sub.1-C.sub.6 alkyl optionally
substituted with --OCH.sub.3, --OCD.sub.3, --OCF.sub.3, or
--OCF.sub.2H; [0181] Q.sup.1 is selected from H, C.sub.1-C.sub.8
alkyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.6-C.sub.10 aryl, 5-6
membered heteroaryl, or 5-6 membered heterocyclyl groups, wherein
when Q.sup.1 is not H, said Q.sup.1 is optionally substituted with
1-4 substituents independently selected from halogen, --OR.sup.6,
--SR.sup.6, --N(R.sup.6).sub.2, C.sub.6-C.sub.10 aryl,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, --NO.sub.2, --CN, --CF.sub.3,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --S(O)(C.sub.1-C.sub.6 alkyl),
--NR.sup.6SO.sub.2Z.sup.2, --SO.sub.2NR.sup.17R.sup.18,
--NHCOOR.sup.16, --NHCOZ.sup.2, --NHCONHR.sup.16,
--CO.sub.2R.sup.6, --C(O)R.sup.6, and --CON(R.sup.6).sub.2; [0182]
Q.sup.2 is C.sub.5-C.sub.10 spiro bicyclic carbocyclyl optionally
substituted with 1-4 Z.sup.1 groups; [0183] Q.sup.3 is
C.sub.5-C.sub.10 fused bicyclic carbocyclyl optionally substituted
with 1-4 Z.sup.1 groups; [0184] Q.sup.4 is C.sub.5-C.sub.10 bridged
bicyclic carbocyclyl optionally substituted with 1-4 Z.sup.1
groups; [0185] Q.sup.5 is 4-membered heterocyclyl having 1
heteroatom selected from N, O or S wherein Q.sup.5 is optionally
substituted with 1-4 Z.sup.3 groups; [0186] Q.sup.6 is
C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.6-C.sub.10 aryl, 5-6 membered heteroaryl, or 5-6 membered
heterocyclyl, wherein Q.sup.6 is substituted with 1 oxo group and
with 0 to 3 substituents independently selected from halogen,
--OR.sup.6, --SR.sup.6, --N(R.sup.6).sub.2, C.sub.6-C.sub.10 aryl,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, --NO.sub.2, --CN, --CF.sub.3,
--SO.sub.2(C.sub.1-C.sub.6 alkyl), --S(O)(C.sub.1-C.sub.6 alkyl),
--NR.sup.6SO.sub.2Z.sup.2, --SO.sub.2NR.sup.17R.sup.18,
--NHCOOR.sup.16, --NHCOZ.sup.2, --NHCONHR.sup.16,
--CO.sub.2R.sup.6, --C(O)R.sup.6, or --CON(R.sup.6).sub.2; [0187]
Q.sup.7 is C.sub.3-C.sub.8 carbocyclyl, wherein Q.sup.7 is
substituted with 4-8 F atoms and each carbon of Q.sup.7 is
substituted with 0-2 F atoms; [0188] each Z.sup.1 is independently
oxo, halogen, C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8 alkenyl,
C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8 haloalkyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10 membered
heterocyclyl, --CN, --C(O)R.sup.16, --C(O)OR.sup.16,
--C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)R.sup.16, --NR.sup.16C(O)NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2R.sup.16,
--NR.sup.16S(O).sub.2NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2OR.sup.16, --OR.sup.16, --OC(O)R.sup.16,
--OC(O)NR.sup.17R.sup.18, --SR.sup.16, --S(O)R.sup.16,
--S(O).sub.2R.sup.16 or --S(O).sub.2NR.sup.17R.sup.18 wherein any
alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl or
heterocyclyl of Z.sup.1 is optionally substituted with 1-4 Z.sup.1a
groups; [0189] each Z.sup.1a is independently oxo, halogen,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8
haloalkyl, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10
membered heterocyclyl, --CN, --C(O)R.sup.16, --C(O)OR.sup.16,
--C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)R.sup.16, --NR.sup.16C(O)OR.sup.16,
--NR.sup.16C(O)NR.sup.17R.sup.18, --NR.sup.16S(O).sub.2R.sup.16,
--NR.sup.16S(O).sub.2NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2OR.sup.16, --OR.sup.16, --OC(O)R.sup.16,
--OC(O)NR.sup.17R.sup.18, --SR.sup.16, --S(O)R.sup.16,
--S(O).sub.2R.sup.16 or --S(O).sub.2NR.sup.17R.sup.18 wherein any
alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclyl of
Z.sup.1a is optionally substituted with 1-4 Z.sup.1c groups; [0190]
each R.sup.16
is independently H, C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8 alkenyl,
C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl or 4-10 membered heterocyclyl, wherein any
alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl or
heterocyclyl of R.sup.16 is optionally substituted with 1-4
Z.sup.1c groups; [0191] each Z.sup.1c is independently oxo,
halogen, C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8 haloalkyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10 membered
heterocyclyl, --CN, --C(O)(C.sub.1-C.sub.8 alkyl),
--C(O)O(C.sub.1-C.sub.8 alkyl), --C(O)N(C.sub.1-C.sub.8
alkyl).sub.2, --NH.sub.2, --NH(C.sub.1-C.sub.8 alkyl),
--N(C.sub.1-C.sub.8 alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.8 alkyl),
--NHC(O)(C.sub.1-C.sub.8 alkyl), --NHC(O)NH(C.sub.1-C.sub.8 alkyl),
--OH, --O(C.sub.1-C.sub.8 alkyl), C.sub.3-C.sub.8 cycloalkoxy,
C.sub.5-C.sub.10 bicyclic carbocyclyloxy, --S(C.sub.1-C.sub.8
alkyl) or --S(O).sub.2N(C.sub.1-C.sub.8 alkyl).sub.2 wherein any
alkyl, carbocyclyl, aryl, heteroaryl, heterocyclyl or cycloalkoxy
portion of Z.sup.1c is optionally substituted with 1-4 halogen
atoms or C.sub.1-C.sub.6 alkoxy groups; [0192] R.sup.17 and
R.sup.18 are each independently H, C.sub.1-C.sub.8 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, --C(O)R.sup.16,
--C(O)OR.sup.16, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or
4-10 membered heterocyclyl, wherein any alkyl, alkenyl, alkynyl,
carbocyclyl, aryl, heteroaryl or heterocyclyl of R.sup.17 or
R.sup.18 is optionally substituted with 1-4 Z.sup.1c groups, or
R.sup.17 and R.sup.18 together with the nitrogen to which they are
attached form a 4-7 membered heterocyclyl group, wherein said 4-7
membered heterocyclyl group is optionally substituted with 1-4
Z.sup.1c groups; [0193] each Z.sup.2 is independently
C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl,
--NR.sup.17R.sup.18 or --OR.sup.16 wherein any alkyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl portion of Z.sup.2 is optionally
substituted with 1-4 Z.sup.2a groups; [0194] each Z.sup.2a is
independently oxo, halogen, C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.5-C.sub.10 bicyclic
carbocyclyl, C.sub.1-C.sub.8 haloalkyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl, --(C.sub.2-C.sub.8
alkynyl)aryl, --(C.sub.2-C.sub.8 alkynyl)heteroaryl, --CN,
--C(O)(C.sub.1-C.sub.6 alkyl), --C(O)O(C.sub.1-C.sub.6 alkyl),
--C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)O(C.sub.1-C.sub.6 alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --OH, --O(C.sub.1-C.sub.6
alkyl), halo(C.sub.1-C.sub.6 alkoxy), C.sub.3-C.sub.8 cycloalkoxy,
--S(C.sub.1-C.sub.6 alkyl), or --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2; wherein any alkyl, alkynyl, carbocyclyl, cycloalkoxy,
aryl, heteroaryl or heterocyclyl portions of Z.sup.2a is optionally
substituted with 1-4 halogen or C.sub.1-C.sub.6 alkoxy groups;
[0195] each Z.sup.3 is independently oxo, halogen, C.sub.2-C.sub.8
alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8 haloalkyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10 membered
heterocyclyl, --CN, --C(O)OR.sup.16, --C(O)NR.sup.17R.sup.18,
--NR.sup.17R.sup.18, --NR.sup.16C(O)NR.sup.17R.sup.18, --OR.sup.16,
--SR.sup.16 or --SO.sub.2R.sup.16; wherein any alkenyl, alkynyl,
carbocyclyl, aryl, heteroaryl or heterocyclyl portions of Z.sup.3
is optionally substituted with 1-4 halogen; and [0196] each Z.sup.4
is independently oxo, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.5-C.sub.10 bicyclic
carbocyclyl, C.sub.1-C.sub.8 haloalkyl, C.sub.8-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl, --CN,
--C(O)OR.sup.16, --C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)NR.sup.17R.sup.18, --OR.sup.16, --SR.sup.16 or
--SO.sub.2R.sup.16, wherein any alkenyl, alkynyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl portions of Z.sup.4 is optionally
substituted with 1-4 halogen.
[0197] In a further embodiment a compound of Formula (III):
##STR00005##
or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof, is provided, wherein:
[0198] M is --O--; [0199] J is J.sup.1, J.sup.2, J.sup.3, J.sup.4,
J.sup.5, J.sup.6, J.sup.7, J.sup.8 or J.sup.9; [0200] {circle
around (T)} is T.sup.1, T.sup.2, T.sup.3, T.sup.4, T.sup.5,
T.sup.7, T.sup.8, T.sup.9, T.sup.10, T.sup.11, T.sup.12, T.sup.13
or T.sup.14; [0201] L is L.sup.1, L.sup.2, L.sup.3, L.sup.4,
L.sup.5, L.sup.6, L.sup.7, L.sup.8, L.sup.9 or L.sup.10; [0202] Q
is Q.sup.1, Q.sup.2, Q.sup.3, Q.sup.4, Q.sup.5 or Q.sup.7; [0203] E
is E.sup.1, E.sup.2, E.sup.3, or E4; [0204] {circle around (U)} is
selected from U.sup.1, U.sup.3, U.sup.4, U.sup.5, U.sup.6 or
U.sup.7; [0205] J.sup.1 is halogen; [0206] J.sup.2 is --OH; [0207]
J.sup.3 is --NR.sup.17R.sup.18; [0208] J.sup.4 is C.sub.1-C.sub.8
alkyl; [0209] J.sup.5 is C.sub.1-C.sub.8 alkyl substituted with 1-4
Z.sup.3 groups; [0210] J.sup.6 is C.sub.3-C.sub.8 carbocyclyl
optionally substituted with 1-4 Z.sup.3 groups; [0211] J.sup.7 is
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, or 4-10 membered
heterocyclyl any of which groups are optionally substituted with
1-4 Z.sup.3 groups; [0212] J.sup.8 is C.sub.1-C.sub.8 alkoxy
optionally substituted with 1-4 Z.sup.3 groups; [0213] J.sup.9 is
C.sub.3-C.sub.8 carbocyclyleoxy optionally substituted with 1-4
Z.sup.3 groups; [0214] T.sup.1 is C.sub.3-C.sub.8 carbocyclylene
attached to L and M through two adjacent carbons; [0215] T.sup.2 is
C.sub.3-C.sub.8 carbocyclylene attached to L and M through two
adjacent carbons, wherein said carbocyclylene is substituted with
1-4 C.sub.1-C.sub.8 alkyl groups; [0216] T.sup.3 is C.sub.3-C.sub.8
carbocyclylene attached to L and M through two adjacent carbons,
wherein said carbocyclylene is substituted with 1-4 halogen atoms
and said carbocyclylene is optionally substituted with 1-4
C.sub.1-C.sub.6 alkyl groups; [0217] T.sup.4 is C.sub.3-C.sub.8
carbocyclylene that is attached to L and M through two adjacent
carbons, wherein said carbocyclylene is substituted with a
C.sub.1-C.sub.8 alkyl group, wherein said alkyl group is
substituted with 1-4 Z.sup.3 groups; [0218] T.sup.5 is 4-10
membered heterocyclene that is attached to L and M through two
adjacent carbons; [0219] T.sup.7 is 4-10 membered heterocyclene
that is attached to M through a carbon atom and attached to L
through a N atom, wherein said heterocyclene is optionally
substituted with 1-4 Z.sup.1 groups; [0220] T.sup.8 is 4-10
membered heterocyclene that is attached to L and M through two
adjacent carbons, wherein said heterocyclene is substituted with
1-4 Z.sup.1 groups; [0221] T.sup.9 is C.sub.5-C.sub.12 spiro
bicyclic carbocyclylene that is attached to L and M through two
adjacent carbons, wherein said spiro bicyclic carbocyclylene is
optionally substituted with 1-4 Z.sup.1 groups; [0222] T.sup.10 is
C.sub.5-C.sub.12 fused bicyclic carbocyclylene that is attached to
L and M through two adjacent carbons, wherein said fused bicyclic
carbocyclylene is optionally substituted with 1-4 Z.sup.1 groups;
[0223] T.sup.11 is C.sub.5-C.sub.12 bridged bicyclic carbocyclylene
that is attached to L and M through two adjacent carbons, wherein
said bridged bicyclic carbocyclylene is optionally substituted with
1-4 Z.sup.1 groups; [0224] T.sup.12 is C.sub.4-C.sub.8
carbocyclylene that is attached to L and M through two non-adjacent
carbons, wherein said carbocyclylene is optionally substituted with
1-4 Z.sup.1 groups; [0225] T.sup.13 is a 5-8 membered fused,
bridged, or spiro bicyclic heterocyclene that is attached to L and
M through two adjacent atoms, wherein said heterocyclene is
optionally substituted with 1-4 Z.sup.1 groups; [0226] T.sup.14 is
C.sub.3-C.sub.8 carbocyclylene that is attached to L and M through
two adjacent carbons, wherein said carbocyclylene is substituted
with 1-4 Z.sup.4 groups; [0227] L.sup.1 is C.sub.1-C.sub.8 alkylene
or C.sub.2-C.sub.8 alkenylene; [0228] L.sup.2 is C.sub.1-C.sub.8
alkylene or C.sub.2-C.sub.8 alkenylene wherein said C.sub.1-C.sub.8
alkylene or said C.sub.2-C.sub.8 alkenylene is substituted with 1-4
halogens; [0229] L.sup.3 is C.sub.1-C.sub.8 alkylene or
C.sub.2-C.sub.8 alkenylene wherein said C.sub.1-C.sub.8 alkylene or
said C.sub.2-C.sub.8 alkenylene and wherein said C.sub.1-C.sub.8
alkylene or said C.sub.2-C.sub.8 alkenylene is optionally
substituted with 1-4 halogens; [0230] L.sup.4 is C.sub.1-C.sub.8
alkylene or C.sub.2-C.sub.8 alkenylene substituted with two geminal
C.sub.1-C.sub.4 alkyl groups that come together to form a spiro
C.sub.3-C.sub.8 carbocyclyl group, wherein L.sup.4 is optionally
substituted with 1-4 Z.sup.1 groups; [0231] L.sup.5 is 2-8 membered
heteroalkylene or 4-8 membered heteroalkenylene that is connected
to {circle around (T)} by an O, S or N atom and said heteroalkylene
or heteroalkenylene is optionally substituted with 1-4 Z.sup.3
groups; [0232] L.sup.6 is 2-8 membered heteroalkylene or 5-8
membered heteroalkenylene that is connected to {circle around (T)}
by a carbon atom and said heteroalkylene or heteroalkenylene is
substituted with 1-4 halogen atoms and is optionally substituted
with 1-4 Z.sup.4 groups; [0233] L.sup.7 is 2-8 membered
heteroalkylene or 4-8 membered heteroalkenylene that is connected
to {circle around (T)} by a carbon atom and said heteroalkylene or
heteroalkenylene is optionally substituted with 1-4 Z.sup.4 groups;
[0234] L.sup.8 is L.sup.8A-L.sup.8B-L.sup.8 wherein L.sup.8A and
L.sup.8C are each independently C.sub.1-C.sub.6 alkylene,
C.sub.1-C.sub.6 heteroalkylene, C.sub.2-C.sub.6 alkenylene or a
bond and L.sup.8B is a 3- to 6-membered saturated or unsaturated
ring containing 0 to 3 heteroatoms selected from N, O, or S,
wherein L.sup.8A and L.sup.8C connect to L.sup.8B at two different
ring atoms and L.sup.8B is optionally substituted with 1-4 Z.sup.1
groups; [0235] L.sup.9 is C.sub.2-C.sub.8 alkynylene optionally
substituted with 1-4 Z.sup.1 groups; [0236] L.sup.10 is
C.sub.1-C.sub.8 alkylene or C.sub.3-C.sub.8 alkenylene substituted
with two geminal Z.sup.1 groups that come together to form a spiro
4-8 membered heterocyclyl group, wherein L.sup.10 is optionally
substituted with 1-4 Z.sup.1 groups; [0237] U.sup.1 is a
C.sub.6-C.sub.14 membered arylene optionally substituted with 1-4 W
groups; [0238] U.sup.3 is a 4-14 membered heterocyclene optionally
substituted with 1-4 W groups that are located on one or more ring
atoms selected from C or N; [0239] U.sup.4 is a 5 or 6 membered
monocyclic heteroarylene containing 1, 2 or 3 heteroatoms
independently selected from N, O, or S, wherein said heteroarylene
is optionally substituted with 1-4 W groups that are located on one
or more ring atoms selected from C or N; [0240] U.sup.5 is a 8, 9
or 10 membered fused bicyclic heteroarylene containing 1, 2 or 3
heteroatom ring atoms independently selected from N, O, or S,
wherein said heteroarylene is optionally substituted with 1-4 W
groups that are located on one or more ring atoms selected from C
or N; [0241] U.sup.6 is a 11-14 membered fused tricyclic
heteroarylene containing 1, 2, 3 or 4 heteroatom ring atoms
independently selected from N, O, or S, wherein said heteroarylene
is optionally substituted with 1-4 W groups that are located on one
or more ring atoms selected from C or N; [0242] U.sup.7 is a 8-10
membered fused bicyclic heteroarylene containing 4 heteroatom ring
atoms independently selected from N, O, or S, wherein said
heteroaryl is optionally substituted with 1-2 W groups that are
located on one or more ring atoms selected from C or N; [0243] each
W is independently W.sup.1, W.sup.2, W.sup.3, W.sup.4, W.sup.5,
W.sup.6 or W.sup.7; [0244] each W.sup.1 is independently oxo,
halogen, --OR.sup.6, C.sub.1-C.sub.6 alkyl, --CN, --CF.sub.3,
--SR.sup.6, --C(O).sub.2R.sup.6, --C(O)N(R.sup.6).sub.2,
--C(O)R.sup.6, --N(R.sup.6)C(O)R.sup.6, --SO.sub.2(C.sub.1-C.sub.6
alkyl), --S(O)(C.sub.1-C.sub.6 alkyl), C.sub.3-C.sub.8 carbocyclyl,
C.sub.3-C.sub.8 cycloalkoxy, C.sub.1-C.sub.6 haloalkyl,
--N(R.sup.6).sub.2, --NR.sup.6(C.sub.1-C.sub.6
alkyl)O(C.sub.1-C.sub.6 alkyl), halo(C.sub.1-C.sub.6 alkoxy),
--NR.sup.6SO.sub.2R.sup.6, --SO.sub.2N(R.sup.6).sub.2,
--NHCOOR.sup.6, --NHCONHR.sup.6, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl or --O(4-10
membered heterocyclyl), wherein said W.sup.1 alkyl, carbocyclyl,
cycloalkoxy, haloalkyl, haloalkoxy, aryl, heteroaryl, or
heterocyclyl is optionally substituted with 1-4 Z.sup.1c groups;
[0245] each R.sup.6 is independently H, C.sub.6-C.sub.10 aryl or
C.sub.1-C.sub.6 alkyl, wherein said aryl or alkyl is optionally
substituted with 1 to 4 substituents independently selected from
halogen atoms, C.sub.1-C.sub.6 alkyl, C.sub.6-C.sub.10 aryl,
C.sub.3-C.sub.8 carbocyclyl, 5-14 membered heteroaryl, 4-10
membered heterocyclyl, halo(C.sub.1-C.sub.6 alkoxy), --OH,
--O(C.sub.1-C.sub.6 alkyl), --SH, --S(C.sub.1-C.sub.6 alkyl),
--NH.sub.2, --NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6
alkyl).sub.2, --C(O)(C.sub.1-C.sub.6 alkyl),
--SO.sub.2N(C.sub.1-C.sub.6 alkyl).sub.2, --NHCOO(C.sub.1-C.sub.6
alkyl), --NHCO(C.sub.1-C.sub.6 alkyl), --NHCONH(C.sub.1-C.sub.6
alkyl), --CO.sub.2(C.sub.1-C.sub.6 alkyl), or
--C(O)N(C.sub.1-C.sub.6 alkyl).sub.2; [0246] each W.sup.2 is
C.sub.1-C.sub.6 alkoxy substituted with a 5-14 membered heteroaryl
or C.sub.6-C.sub.10 aryl; wherein said heteroaryl or aryl is
substituted with 1-4 Z.sup.1 groups; [0247] each W.sup.3 is
C.sub.2-C.sub.6 alkynyl group substituted with a C.sub.6-C.sub.10
aryl, C.sub.3-C.sub.8 carbocyclyl, C.sub.1-C.sub.8 alkyl,
C.sub.1-C.sub.6 haloalkyl, 4-10 membered heterocyclyl, or 5-14
membered heteroaryl group; wherein said aryl, carbocyclyl, alkyl,
haloalkyl, heterocyclyl, or heteroaryl group is optionally
substituted with 1-4 Z.sup.1 groups; [0248] each W.sup.4 is
--SF.sub.5; [0249] each W.sup.5 is --O(C.sub.2-C.sub.6
alkyl)OR.sup.22 wherein R.sup.22 is a C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl or 4-10 membered heterocyclyl group optionally
substituted with 1-4 Z.sup.1 groups; [0250] each W.sup.6 is
--O(C.sub.2-C.sub.6 alkyl)NR.sup.16R.sup.22 wherein R.sup.22 is an
aryl, heteroaryl or heterocyclyl group optionally substituted with
1-4 Z.sup.1 groups; [0251] each W.sup.7 is --O(5-14 membered
heteroaryl); wherein said --O(5-14 membered heteroaryl) is
optionally substituted with 1-4 Z.sup.1 groups and 2 adjacent
substituents of said --O(5-14 membered heteroaryl) may be taken
together to form a 3- to 6-membered cyclic ring containing 0 to 3
heteroatoms independently selected from N, O, or S; [0252] E is
[0252] ##STR00006## [0253] E.sup.2 is
[0253] ##STR00007## [0254] E.sup.3 is
[0254] ##STR00008## [0255] E.sup.4 is
[0255] ##STR00009## [0256] Q.sup.1 is H, C.sub.1-C.sub.8 alkyl,
C.sub.3-C.sub.8 carbocyclyl, C.sub.6-C.sub.10 aryl, 5-6 membered
heteroaryl, or 5-6 membered heterocyclyl groups, wherein when
Q.sup.1 is not H, said Q.sup.1 is optionally substituted with 1-4
substituents independently selected from halogen, --OR.sup.6,
--SR.sup.6, --N(R.sup.6).sub.2, C.sub.6-C.sub.10 aryl,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, --CN, --CF.sub.3, --SO.sub.2(C.sub.1-C.sub.6 alkyl),
--S(O)(C.sub.1-C.sub.6 alkyl), --NR.sup.6SO.sub.2Z.sup.2,
--SO.sub.2NR.sup.17R.sup.18, --NHCOOR.sup.16, --NHCOZ.sup.2,
--NHCONHR.sup.16, --CO.sub.2R.sup.6, --C(O)R.sup.6 or
--CON(R.sup.6).sub.2; [0257] Q.sup.2 is C.sub.5-C.sub.10 spiro
bicyclic carbocyclyl optionally substituted with 1-4 Z.sup.1
groups; [0258] Q.sup.3 is C.sub.5-C.sub.10 fused bicyclic
carbocyclyl optionally substituted with 1-4 Z.sup.1 groups; [0259]
Q.sup.4 is C.sub.5-C.sub.10 bridged bicyclic carbocyclyl optionally
substituted with 1-4 Z.sup.1 groups; [0260] Q.sup.5 is 4-membered
heterocyclyl having 1 heteroatom selected from N, O or S wherein
Q.sup.5 is optionally substituted with 1-4 Z.sup.3 groups; [0261]
Q.sup.7 is C.sub.3-C.sub.8 carbocyclyl substituted with 4-8 F atoms
and each carbon of Q.sup.7 is substituted with 0-2 F atoms; [0262]
each Z.sup.1 is independently oxo, halogen, C.sub.1-C.sub.8 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8
haloalkyl, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10
membered heterocyclyl, --CN, --C(O)R.sup.16, --C(O)OR.sup.16,
--C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)R.sup.16, --NR.sup.16C(O)NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2R.sup.16,
--NR.sup.16S(O).sub.2NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2OR.sup.16, --OR.sup.16, --OC(O)R.sup.16,
--OC(O)NR.sup.17R.sup.18, --SR.sup.16, --S(O)R.sup.16,
--S(O).sub.2R.sup.16 or --S(O).sub.2NR.sup.17R.sup.18 wherein any
alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl or
heterocyclyl of Z.sup.1 is optionally substituted with 1-4 Z.sup.1a
groups; [0263] each Z.sup.1a is independently oxo, halogen,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8
haloalkyl, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10
membered heterocyclyl, --CN, --C(O)R.sup.16, --C(O)OR.sup.16,
--C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)R.sup.16, --NR.sup.16C(O)OR.sup.16,
--NR.sup.16C(O)NR.sup.17R.sup.18, --NR.sup.16S(O).sub.2R.sup.16,
--NR.sup.16S(O).sub.2NR.sup.17R.sup.18,
--NR.sup.16S(O).sub.2OR.sup.16, --OR.sup.16, --OC(O)R.sup.16,
--OC(O)NR.sup.17R.sup.18, --SR.sup.16, --S(O)R.sup.16,
--S(O).sub.2R.sup.16 or --S(O).sub.2NR.sup.17R.sup.18 wherein any
alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclyl of
Z.sup.1a is optionally substituted with 1-4 Z.sup.1c groups; [0264]
each R.sup.16 is independently H, C.sub.1-C.sub.8 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or 4-10 membered
heterocyclyl, wherein any alkyl, alkenyl, alkynyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl of R.sup.16 is optionally
substituted with 1-4 Z.sup.1c groups; [0265] each Z.sup.1c is
independently oxo, halogen, C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8
haloalkyl, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10
membered heterocyclyl, --CN, --C(O)(C.sub.1-C.sub.8 alkyl),
--C(O)O(C.sub.1-C.sub.8 alkyl), --C(O)N(C.sub.1-C.sub.8
alkyl).sub.2, --NH.sub.2, --NH(C.sub.1-C.sub.8 alkyl),
--N(C.sub.1-C.sub.8 alkyl).sub.2, --NHC(O)O(C.sub.1-C.sub.8 alkyl),
--NHC(O)(C.sub.1-C.sub.8 alkyl), --NHC(O)NH(C.sub.1-C.sub.8 alkyl),
--OH, --O(C.sub.1-C.sub.8 alkyl), C.sub.3-C.sub.8 cycloalkoxy,
C.sub.5-C.sub.10 bicyclic carbocyclyloxy, --S(C.sub.1-C.sub.8
alkyl) or --S(O).sub.2N(C.sub.1-C.sub.8 alkyl).sub.2 wherein any
alkyl, carbocyclyl, aryl, heteroaryl, heterocyclyl or cycloalkoxy
portion of Z.sup.1c is optionally substituted with 1-4 halogen
atoms or C.sub.1-C.sub.6 alkoxy groups; [0266] R.sup.17 and
R.sup.18 are each independently H, C.sub.1-C.sub.8 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8
carbocyclyl, C.sub.5-C.sub.10 bicyclic carbocyclyl, --C(O)R.sup.16,
--C(O)OR.sup.16, C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl or
4-10 membered heterocyclyl, wherein any alkyl, alkenyl, alkynyl,
carbocyclyl, aryl, heteroaryl or heterocyclyl of R.sup.17 or
R.sup.18 is optionally substituted with 1-4 Z.sup.1c groups, or
R.sup.17 and R.sup.18 together with the nitrogen to which they are
attached form a 4-7 membered heterocyclyl group, wherein said 4-7
membered heterocyclyl group is optionally substituted with 1-4
Z.sup.1c groups; [0267] each Z.sup.2 is independently
C.sub.1-C.sub.8 alkyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl,
--NR.sup.17R.sup.18 or --OR.sup.16 wherein any alkyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl portion of Z.sup.2 is optionally
substituted with 1-4 Z.sup.2a groups; [0268] each Z.sup.2a is
independently oxo, halogen, C.sub.1-C.sub.8 alkyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.5-C.sub.10 bicyclic
carbocyclyl, C.sub.1-C.sub.8 haloalkyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl, --(C.sub.2-C.sub.8
alkynyl)aryl, --(C.sub.2-C.sub.8 alkynyl)heteroaryl, --CN,
--C(O)(C.sub.1-C.sub.6 alkyl), --C(O)O(C.sub.1-C.sub.6 alkyl),
--C(O)N(C.sub.1-C.sub.6 alkyl).sub.2, --NH.sub.2,
--NH(C.sub.1-C.sub.6 alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHC(O)O(C.sub.1-C.sub.6 alkyl), --NHC(O)(C.sub.1-C.sub.6 alkyl),
--NHC(O)NH(C.sub.1-C.sub.6 alkyl), --OH, --O(C.sub.1-C.sub.6
alkyl), halo(C.sub.1-C.sub.6 alkoxy), C.sub.3-C.sub.8 cycloalkoxy,
--S(C.sub.1-C.sub.6 alkyl), or --SO.sub.2N(C.sub.1-C.sub.6
alkyl).sub.2; wherein any alkyl, alkynyl, carbocyclyl, cycloalkoxy,
aryl, heteroaryl or heterocyclyl portions of Z.sup.2a is optionally
substituted with 1-4 halogen or C.sub.1-C.sub.6 alkoxy groups;
[0269] each Z.sup.3 is independently oxo, halogen, C.sub.2-C.sub.8
alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.3-C.sub.8 carbocyclyl,
C.sub.5-C.sub.10 bicyclic carbocyclyl, C.sub.1-C.sub.8 haloalkyl,
C.sub.6-C.sub.10 aryl, 5-14 membered heteroaryl, 4-10 membered
heterocyclyl, --CN, --C(O)OR.sup.16, --C(O)NR.sup.17R.sup.18,
--NR.sup.17R.sup.18, --NR.sup.16C(O)NR.sup.17R.sup.18, --OR.sup.16,
--SR.sup.16 or --SO.sub.2R.sup.16; wherein any alkenyl, alkynyl,
carbocyclyl, aryl, heteroaryl or heterocyclyl portions of Z.sup.3
is optionally substituted with 1-4 halogen; and [0270] each Z.sup.4
is independently oxo, C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8
alkynyl, C.sub.3-C.sub.8 carbocyclyl, C.sub.5-C.sub.10 bicyclic
carbocyclyl, C.sub.1-C.sub.8 haloalkyl, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl, --CN,
--C(O)OR.sup.16, --C(O)NR.sup.17R.sup.18, --NR.sup.17R.sup.18,
--NR.sup.16C(O)NR.sup.17R.sup.18, --OR.sup.16, --SR.sup.16 or
--SO.sub.2R.sup.16, wherein any alkenyl, alkynyl, carbocyclyl,
aryl, heteroaryl or heterocyclyl portions of Z.sup.4 is optionally
substituted with 1-4 halogen.
[0271] One embodiment provides a pharmaceutical composition
comprising a compound of Formula I, II, III, or IV (such as any one
of IVa-IVh), or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0272] One embodiment provides a method for treating a Flaviviridae
viral infection (e.g., an HCV viral infection) in a patient in need
thereof (e.g., mammal such as a human). The method includes
administering a compound of Formula I, II, III, or IV (such as any
one of IVa-IVh), or a stereoisomer, or a mixture of stereoisomers,
or a pharmaceutically acceptable salt thereof, to the patient.
[0273] One embodiment provides a method for inhibiting the
proliferation of the HCV virus, treating HCV or delaying the onset
of HCV symptoms in a patient in need thereof (e.g., mammal such as
a human). The method includes administering a compound of Formula
I, II, III, or IV (such as any one of IVa-IVh) or a stereoisomer,
or a mixture of stereoisomers, or a pharmaceutically acceptable
salt thereof, to the patient.
[0274] One embodiment provides a compound of Formula I, II, III, or
IV (such as any one of IVa-IVh) or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof for
use in medical therapy (e.g., for use in treating a Flaviviridae
viral infection such as an HCV viral infection or in treating the
proliferation of the HCV virus or delaying the onset of HCV
symptoms in a patient in need thereof (e.g., mammal such as a
human)).
[0275] One embodiment provides a compound of Formula I, II, III, or
IV (such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof for
use in the manufacture of a medicament for treating a Flaviviridae
viral infection (e.g., an HCV viral infection) or the proliferation
of the HCV virus or delaying the onset of HCV symptoms in a patient
in need thereof (e.g., mammal such as a human).
[0276] One embodiment provides a compound of Formula I, II, III, or
IV (such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, for
use in the prophylactic or therapeutic treatment of the
proliferation of a Flaviviridae virus, an HCV virus or for use in
the therapeutic treatment of delaying the onset of HCV
symptoms.
[0277] One embodiment provides a compound of Formula I, II, III, or
IV (such as any one of IVa-IVh) or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, for
use in the prophylactic or therapeutic treatment of a Flaviviridae
virus infection (e.g., an HCV virus infection).
[0278] One embodiment provides the use of a compound of Formula I,
II, III, or IV (such as any one of IVa-IVh), or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, for the manufacture of a medicament for a Flaviviridae
virus infection (e.g., an HCV virus infection) in a patient in need
thereof (e.g., mammal such as a human).
[0279] One embodiment provides processes and intermediates
disclosed herein that are useful for preparing compounds of Formula
I, II, III, or IV (such as any one of IVa-IVh) or a stereoisomer,
or a mixture of stereoisomers, or salts thereof.
[0280] Other embodiments, objects, features and advantages will be
set forth in the detailed description of the embodiments that
follows, and in part will be apparent from the description, or may
be learned by practice, of the claimed invention. These objects and
advantages will be realized and attained by the processes and
compositions particularly pointed out in the written description
and claims hereof. The foregoing Summary has been made with the
understanding that it is to be considered as a brief and general
synopsis of some of the embodiments disclosed herein, is provided
solely for the benefit and convenience of the reader, and is not
intended to limit in any manner the scope, or range of equivalents,
to which the appended claims are lawfully entitled.
DETAILED DESCRIPTION
[0281] While the present invention is capable of being embodied in
various forms, the description below of several embodiments is made
with the understanding that the present disclosure is to be
considered as an exemplification of the claimed subject matter, and
is not intended to limit the appended claims to the specific
embodiments illustrated. The headings used throughout this
disclosure are provided for convenience only and are not to be
construed to limit the claims in any way. Embodiments illustrated
under any heading may be combined with embodiments illustrated
under any other heading.
ABBREVIATIONS
[0282] The following abbreviations are used throughout the
specification, and have the following meanings: [0283] .degree.
C.=degrees Celsius [0284] A=Angstrom [0285] Ac=acetyl [0286]
AcOH=acetic acid [0287] aq=aqueous [0288] Ar=argon [0289]
atm=atmosphere [0290] BEP=2-bromo-1-ethyl pyridinium
tetrafluoroborate [0291]
Bis(diphenylphosphino)ferrocene)palladium(II) dichloride [0292]
Bn=benzyl [0293] Boc=tert-butoxy carbonyl [0294]
Boc.sub.2O=di-tert-butyl dicarbonate [0295] bp=boiling point [0296]
Bs=4-bromophenylsulfonyl [0297] Bu=butyl [0298] calcd=calculated
[0299] CBS=Corey-Bakshi-Shibata [0300] CBZ=Cbz=carboxybenzyl [0301]
CDI=1,1'-carbonyldiimidazole [0302] cm=centermeter [0303]
COMU=(1-cyano-2-ethoxy-2-oxoethylidenam inooxy)dimethylam
ino-morpholino-carbenium hexafluorophosphate [0304]
DABCO=1,4-diazabicyclo[2.2.2]octane [0305]
DBU=1,8-diazabicycloundec-7-ene [0306] DCE=1,2-dichloroethane
[0307] DCM=dichloromethane [0308] DIAD=diisopropyl azodicarboxylate
[0309] dioxane=1,4-dioxane [0310]
DIPEA=N,N-diisopropyl-N-ethylamine [0311] DMF=N,N-dimethylformamide
[0312] DMAP=4-dimethylaminopyridine [0313]
DMPU=1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone [0314]
DMSO=dimethysulfoxide [0315]
dppf=1,1'-bis(diphenylphosphino)ferrocene [0316]
DSC=N,N'-disuccinimidyl carbonate [0317] EA=EtOAc=ethyl acetate
[0318] EC.sub.50=half maximal effective concentration [0319]
EDC=1-ethyl-3-(3-dimethylaminopropyl)carbodiimide [0320] Et=ethyl
[0321] Et.sub.2O=diethyl ether [0322] EtOAc=ethyl acetate [0323]
EtOH=ethanol [0324] equiv=equivalent [0325] F-NMR=fluorine nuclear
magnetic resonance spectroscopy [0326] g=gram [0327] h=hour [0328]
HATU=O-(7-azabenzotriazol-1-yl)-N,N,N,N'-tetramethyluronium
Hexafluorophosphate [0329] HCV=hepatitis C virus [0330]
Hex=hex=hexanes [0331] HMDS=hexamethyldisilazane(azide) [0332]
HMPA=hexamethylphosphoramide [0333] .sup.1H-NMR=proton nuclear
magnetic resonance spectroscopy [0334] HOAc=acetic acid [0335]
HOBT=hydroxybenzotriazole [0336] HPLC=high pressure liquid
chromatography [0337] Hz=Hertz [0338] IPA=isopropyl alcohol [0339]
i=iso [0340] J=coupling constant [0341] KHMDS=potassium
bis(trimethylsilyl)amide [0342] L=liter [0343] LCMS-ESI=liquid
chromatography mass spectrometer (electrospray ionization) [0344]
LiHMDS=lithium bis(trimethylsilyl)amide [0345] M=molar
concentration (mol/L) [0346] Me=methyl [0347] MeCN=acetonitrile
[0348] MeOH=methanol [0349] MeTHF=2-methyltetrahydrofuran [0350]
mg=milligram [0351] MHz=mega Hertz [0352] mL=milliliter [0353]
mmol=millimole [0354] min=minute [0355] MTBE=methyl tert-butylether
[0356] Ms=methanesulfonyl [0357] MsCl=methanesulfonyl chloride
[0358] MS=molecular sieves [0359] MSA=methylsulfonic acid [0360]
n=normal [0361] N=normal concentration [0362]
NCS=N-chlorosuccinimide [0363] NMM=N-methylmorpholine [0364]
NMO=N-methylmorpholine-N-oxide [0365] NMP=N-methylpyrrolidinone
[0366] o/n=overnight [0367] Pf=9-phenyl-9H-fluoren-9-yl [0368]
PG=protecting group [0369] PE=petroleum ether [0370] Ph=phenyl
[0371] PhMe=toluene [0372] pM=picomolar [0373] PMB=4-methoxybenzyl
[0374] Pr=propyl [0375]
Pd(dppf)Cl.sub.2.dbd.PdCl.sub.2(dppf)=PdCl.sub.2dppf=(1,1'-bis(diphenylph-
osphino)ferrocene)dichloropalladium(II) [0376]
PPh.sub.3=triphenylphosphine [0377] rt=room temperature [0378]
sat=sat.=saturated [0379] sec=secondary [0380]
S.sub.N1=nucleophilic substitution unimolecular [0381]
S.sub.N2=nucleophilic substitution bimolecular [0382]
S.sub.NAr=nucleophilic substitution aromatic [0383] t=tert=tertiary
[0384] TBAF=tetra-n-butylammonium fluoride [0385]
TBS=TBDMS=tert-Butyldimethylsilyl [0386]
TBTU=O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate [0387] TEA=triethylamine [0388] temp=temperature
[0389] TEMPO=(2,2,6,6-Tetramethylpiperidin-1-yl)oxyl [0390]
Tf=trifluoromethanesulfonyl [0391] TFA=trifluoroacetic acid [0392]
THF=tetrahydrofuran [0393] TIPS=triisoproylsilyl [0394] TLC=thin
layer chromatography [0395] TMS=trimethylsilyl [0396]
TMSOTf=trimethylsilyl trifluoromethanesulfonate [0397]
TPAP=tetrapropylammonium perruthenate [0398] Tr=triphenylmethyl
[0399] Ts=para-toluenesulfonyl [0400] w/w=weight/weight ratio
DEFINITIONS
[0401] Unless stated otherwise, the following terms and phrases as
used herein are intended to have the following meanings:
[0402] When a cyclic group (e.g. cycloalkyl, carbocyclyl, bicyclic
carbocyclyl, heteroaryl, heterocyclyl) is limited by a number or
range of numbers, the number or numbers refer to the number of
atoms making up the cyclic group, including any heteroatoms.
Therefore, for example, a 4-8 membered heterocyclyl group has 4, 5,
6, 7 or 8 ring atoms.
[0403] "Alkenyl" refers to a straight or branched chain hydrocarbyl
with at least one site of unsaturation, e.g., a
(sp.sup.2)carbon-(sp.sup.2)carbon double bond. For example, an
alkenyl group can have 2 to 8 carbon atoms (i.e., C.sub.2-C.sub.8
alkenyl), or 2 to 6 carbon atoms (i.e., C.sub.2-C.sub.6 alkenyl).
Examples of suitable alkenyl groups include, but are not limited
to, ethylene or vinyl (--CH.dbd.CH.sub.2) and allyl
(--CH.sub.2CH.dbd.CH.sub.2).
[0404] "Alkenylene" refers to an alkene having two monovalent
radical centers derived by the removal of two hydrogen atoms from
the same or two different carbon atoms of a parent alkene.
Exemplary alkenylene radicals include, but are not limited to,
1,2-ethenylene (--CH.dbd.CH--) or prop-1-enylene
(--CH.sub.2CH.dbd.CH--).
[0405] "Alkoxy" is RO-- where R is alkyl, as defined herein.
Non-limiting examples of alkoxy groups include methoxy, ethoxy and
propoxy.
[0406] "Alkyl" refers to a saturated, straight or branched chain
hydrocarbyl radical. For example, an alkyl group can have 1 to 8
carbon atoms (i.e., (C.sub.1-C.sub.8)alkyl) or 1 to 6 carbon atoms
(i.e., (C.sub.1-C.sub.6 alkyl) or 1 to 4 carbon atoms. Examples of
alkyl groups include, but are not limited to, methyl, ethyl,
propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl,
octyl, nonyl and decyl.
[0407] "Alkylene" refers to an alkyl having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent alkane. Examples of
alkylene radicals include, but are not limited to, methylene
(--CH.sub.2--), ethylene (--CH.sub.2CH.sub.2--), propylene
(--CH.sub.2CH.sub.2CH.sub.2--) and butylene
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--).
[0408] "Alkynyl" refers to a straight or branched chain hydrocarbon
with at least one site of unsaturation, e.g., a
(sp)carbon-(sp)carbon triple bond. For example, an alkynyl group
can have 2 to 8 carbon atoms (C.sub.2-C.sub.8 alkyne) or 2 to 6
carbon atoms (C.sub.2-C.sub.6 alkynyl). Examples of alkynyl groups
include, but are not limited to, acetylenyl (--C.ident.CH) and
propargyl (--CH.sub.2C.ident.CH) groups.
[0409] "Alkynylene" refers to an alkynyl having two monovalent
radical centers derived by the removal of two hydrogen atoms from
the same or two different carbon atoms of a parent alkyne. Typical
alkynylene radicals include, but are not limited to, acetylene
(--C.ident.C--), propargylene (--CH.sub.2C.ident.C--), and
1-pentynylene (--CH.sub.2CH.sub.2CH.sub.2C.ident.C--).
[0410] "Aryl" refers to a single all carbon aromatic ring or a
multiple condensed all carbon ring system (e.g., a fused
multicyclic ring system) wherein at least one of the rings is
aromatic. For example, an aryl group can have 6 to 20 carbon atoms,
6 to 14 carbon atoms, or 6 to 12 carbon atoms. It is to be
understood that the point of attachment of a multiple condensed
ring system, as defined above, can be at any position of the ring
system including an aromatic or a carbocyclyl portion of the ring.
Examples of aryl groups include, but are not limited to, phenyl,
naphthyl, tetrahydronaphthyl and indanyl.
[0411] "Arylene" refers to an aryl as defined herein having two
monovalent radical centers derived by the removal of two hydrogen
atoms from two different carbon atoms of a parent aryl. Typical
arylene radicals include, but are not limited to, phenylene,
e.g.,
##STR00010##
and naphthylene, e.g.,
##STR00011##
[0412] "Bicyclic carbocyclyl" refers to a 5-14 membered saturated
or partially unsaturated bicyclic fused, bridged, or spiro ring
hydrocarbon attached via a ring carbon. In a spiro bicyclic
carbocyclyl, the two rings share a single common carbon atom. In a
fused bicyclic carbocyclyl, the two rings share two common and
adjacent carbon atoms. In a bridged bicyclic carbocyclyl, the two
rings share three or more common, non-adjacent carbon atoms.
Examples of bicyclic carbocyclyl groups include, but are not
limited to spiro bicyclic carbocyclyl groups wherein two
carbocyclyl rings share one common atom
##STR00012##
fused bicyclic carbocyclyl groups wherein two carbocyclyl rings
share two common atoms
##STR00013##
and bridged bicyclic carbocyclyl groups wherein two carbocyclyl
rings share three or more (such as 3, 4, 5 or 6) common atoms
##STR00014##
[0413] "Bicyclic carbocyclylene" refers to a bicyclic carbocyclyl,
as defined above, having two monovalent radical centers derived
from the removal of two hydrogen atoms from the same or two
different carbon atom of a parent bicyclic carbocyclyl. Examples of
bicyclic carbocyclylene groups include, but are not limited to,
spiro bicyclic carbocyclylene groups wherein two carbocyclyl rings
share one common atom
##STR00015##
fused bicyclic carbocyclylene groups wherein two carbocyclyl rings
share two common atoms
##STR00016##
and bridged bicyclic carbocyclylene groups wherein two carbocyclyl
rings share three or more (such as 3, 4, 5 or 6) common atoms
##STR00017##
[0414] "Carbocyclyloxy" is RO-- where R is carbocyclyl, as defined
herein.
[0415] "Bicyclic carbocyclyloxy" is RO-- where R is bicyclic
carbocyclyl, as defined herein.
[0416] "Carbocyclyl", and "carbocycle" refers to a hydrocarbyl
group containing one saturated or partially unsaturated ring
structure, attached via a ring carbon. In various embodiments,
carbocyclyl refers to a saturated or a partially unsaturated
C.sub.3-C.sub.12 cyclic moiety, examples of which include
cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,
cyclohexenyl, cycloheptyl and cyclooctyl.
[0417] "Carbocyclylene" (as well as "carbocyclene") refers to a
carbocyclyl, as defined herein, having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent carbocyclyl. Examples of
carbocyclene include, but are not limited to, cyclopropylene,
cyclobutylene, cyclopentylene and cyclohexylene.
[0418] "Carbocyclylalkyl" refers to a hydrocarbyl group containing
one saturated or partially unsaturated ring structure attached to
an alkyl group, attached via a ring carbon or an alkyl carbon. In
various embodiments, carbocyclylalkyl refers to a saturated or a
partially unsaturated C.sub.1-C.sub.12 carbocyclylalkyl moiety,
examples of which include cyclopropylalkyl, cyclobutylalkyl,
cyclopropylethyl, and cyclopropylpropyl.
[0419] "Carbocyclylalkylene" refers to a carbocyclylalkyl, as
defined herein, having two monovalent radical centers derived by
the removal of two hydrogen atoms from the same or two different
carbon atoms of a parent cycloalkylalkyl. Examples of cycloalkylene
include, but are not limited to, cyclopropylmethylene and
cyclopropylmethylene.
[0420] "Cycloalkyl" refers to a hydrocarbyl group containing one
saturated ring structure, attached via a ring carbon. In various
embodiments, cycloalkyl refers to a saturated C.sub.3-C.sub.12
cyclic moiety, examples of which include cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
[0421] "Cycloalkoxy" is RO-- where R is cycloalkyl, as defined
herein.
[0422] "Direct bond" refers a covalent bond between two atoms.
[0423] "Halo" or "halogen" refers to chloro (--Cl), bromo (--Br),
fluoro (--F) or iodo (--I).
[0424] "Haloalkenyl" refers to alkenyl group, as defined herein,
substituted with one or more halogen atoms.
[0425] "Haloalkoxy" refers to alkoxy, as defined herein,
substituted with one or more halogen atoms.
[0426] "Haloalkyl" refers to an alkyl group, in which one or more
hydrogen atoms of the alkyl group is replaced with a halogen atom.
Examples of haloalkyl groups include, but are not limited to,
--CF.sub.3, --CHF.sub.2, --CFH.sub.2 and --CH.sub.2CF.sub.3.
[0427] "Haloalkylene" refers to alkylene group, as defined herein,
substituted with one or more halogen atoms.
[0428] "Heteroalkyl" refers to an alkyl group, as defined herein,
in which one or more carbon atoms is replaced with an oxygen,
sulfur, or nitrogen atom.
[0429] "Heteroalkylene" refers to an alkylene group, as defined
herein, in which one or more carbon atoms is replaced with an
oxygen, sulfur, or nitrogen atom.
[0430] "Heteroalkenyl" refers to an alkenyl group, as defined
herein, in which one or more carbon atoms is replaced with an
oxygen, sulfur, or nitrogen atom.
[0431] "Heteroalkenylene" refers to heteroalkenyl group, as defined
above, having two monovalent radical centers derived by the removal
of two hydrogen atoms from the same or two different atoms of a
parent heteroalkenyl group.
[0432] "Heteroaryl" refers to a single aromatic ring that has at
least one atom other than carbon in the ring, wherein the atom is
selected from the group consisting of oxygen, nitrogen and sulfur;
the term also includes multiple condensed ring systems that have at
least one such aromatic ring. For example, heteroaryl includes
monocyclic, bicyclic or tricyclic ring having up to 6 atoms in each
ring, wherein at least one ring is aromatic and contains from 1 to
4 heteroatoms in the ring selected from the group consisting of
oxygen, nitrogen and sulfur. The rings of the multiple condensed
ring system can be connected to each other via fused, spiro and
bridged bonds when allowed by valency requirements. Non-limiting
examples of heteroaryl include pyridyl, thienyl, furanyl,
pyrimidyl, imidazolyl, pyranyl, pyrazolyl, thiazolyl, thiadiazolyl,
isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl, pyridazinyl,
pyrazinyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzofuranyl,
dibenzofuranyl, dibenzothiophenyl, benzothienyl, indolyl,
benzothiazolyl, benzooxazolyl, benzimidazolyl, isoindolyl,
benzotriazolyl, purinyl, thianaphthenyl and pyrazinyl. Attachment
of heteroaryl can occur via an aromatic ring, or, if heteroaryl is
bicyclic or tricyclic and one of the rings is not aromatic or
contains no heteroatoms, through a non-aromatic ring or a ring
containing no heteroatoms. "Heteroaryl" is also understood to
include the N-oxide derivative of any nitrogen containing
heteroaryl.
[0433] "Heteroarylene" refers to a heteroaryl, as defined above,
having two monovalent radical centers derived by the removal of two
hydrogen atoms from the same or two different carbon atoms or the
removal of a hydrogen from one carbon atom and the removal of a
hydrogen atom from one nitrogen atom of a parent heteroaryl group.
Non-limiting examples of heteroarylene groups are:
##STR00018##
[0434] "Heterocyclyl" refers to a saturated or partially
unsaturated monocyclic, bicyclic or tricyclic group of 2 to 14
ring-carbon atoms and, in addition to ring-carbon atoms, 1 to 4
heteroatoms selected from nitrogen, oxygen and sulfer. Bi- or
tricyclic heterocyclyl groups may have fused, bridged, or spiro
ring connectivity. In various embodiments the heterocyclic group is
attached to another moiety through carbon or through a heteroatom.
Examples of heterocyclyl include without limitation azetidinyl,
oxazolinyl, isoxazolinyl, oxetanyl, tetrahydropyranyl,
tetrahydrothiopyranyl, tetrahydroisoquinolinyl, 1,4-dioxanyl,
pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzoimidazolyl,
dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl,
dihydrofuranyl, dihydroimidazolyl, dihydroindolyl,
dihydroisooxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl,
dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl,
dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl,
dihydroquinolinyl, dihydrotetrazolyl, dihydrothiadiazolyl,
dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl,
dihydroazetidinyl, methylenedioxybenzoyl, chromanyl,
dihydropyranoquinoxalinyl, tetrahydroquinoxalinyl,
tetrahydroquinolinyl, dihydropyranoquinolinyl and tetrahydrothienyl
and N-oxides thereof. A spiro bicyclic heterocyclyl group refers to
a bicyclic heterocyclyl group wherein the two rings of the bicyclic
heterocyclyl group share one common atom. A fused bicyclic bicyclic
heterocyclyl group refers to a bicyclic heterocyclyl group wherein
the two rings of the bicyclic heterocyclyl group share two common
atoms. A bridged bicyclic heterocyclyl group refers to a bicyclic
heterocyclyl group wherein the two rings of the bicyclic
heterocyclyl group share three or more (such as 3, 4, 5 or 6)
common atoms.
[0435] "Heterocyclene" refers to a heterocyclyl, as defined herein,
having two monovalent radical centers derived from the removal of
two hydrogen atoms from the same or two different carbon atoms,
through a carbon and a heteroatom, or through two heteroatoms of a
parent heterocycle.
[0436] "Prodrug" refers to any compound that when administered to a
biological system generates the drug substance, or active
ingredient, as a result of spontaneous chemical reaction(s), enzyme
catalyzed chemical reaction(s), photolysis, and/or metabolic
chemical reaction(s). A prodrug is thus a covalently modified
analog or latent form of a therapeutically active compound.
Non-limiting examples of prodrugs include ester moieties,
quaternary ammonium moieties, glycol moieties, and the like.
[0437] The term "optionally substituted" refers to a moiety wherein
all substituents are hydrogen or wherein one or more of the
hydrogens of the moiety are replaced by non-hydrogen substituents;
that is to say the moiety that is optionally substituted is either
substituted or unsubstituted.
[0438] "Leaving group" (LG) refers to a moiety of a compound that
is active towards displacement or substitution in a chemical
reaction. Examples of in which such as displacement or substitution
occur include, but are not limited to, nucleophilic substitution
bimolecular (S.sub.N2), nucleophilic substitution unimolecular
(S.sub.N1), nucleophilic aromatic substitution (S.sub.NAr), and
transition metal catalyzed cross-couplings. Examples of leaving
groups include, but are not limited to, a halogen atom (e.g. --Cl,
--Br, --I) and sulfonates (e.g. mesylate (--OMs), tosylate (--OTs)
or triflate (--OTf)). The skilled artisan will be aware of various
chemical leaving groups and strategies for activation and will
appreciate the appropriate moiety that will act as leaving groups,
based on the particular chemical reaction, the functionality that
the group is attached to, and the chemical reagents used to affect
the displacement or substitution reaction. As a non-limiting
example, in some situations, a halogen atom (e.g. --Cl, --Br, or
--I) serves as a leaving group in a reaction catalyzed by a
transition metal (e.g. Pd catalyzed Suzuki coupling between an aryl
halide and aryl boronic acid) and another reagents such as a
base.
Stereoisomers
[0439] Stereochemical definitions and conventions used herein
generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of
Chemical Terms (1984) McGraw-Hill Book Company, New York; and
Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds
(1994) John Wiley & Sons, Inc., New York.
[0440] The term "chiral" refers to molecules which have the
property of non-superimposability of the mirror image partner,
while the term "achiral" refers to molecules which are
superimposable on their mirror image partner.
[0441] "Isomers" are different compounds that have the same
molecular formula. Isomers include stereoisomers, enantiomers and
diastereomers.
[0442] "Diastereoisomers" are stereoisomers that have at least two
asymmetric atoms, but which are not mirror-images of each
other.
[0443] "Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term "(.+-.)" is
used to designate a racemic mixture where appropriate.
[0444] The term "stereoisomers" refers to compounds which have
identical chemical constitution, but differ with regard to the
arrangement of the atoms or groups in space.
[0445] The compounds disclosed herein may have chiral centers,
e.g., chiral carbon atoms. Such compounds thus include racemic
mixtures of all stereoisomers, including enantiomers,
diastereomers, and atropisomers. In addition, the compounds
disclosed herein include enriched or resolved optical isomers at
any or all asymmetric, chiral atoms. In other words, the chiral
centers apparent from the depictions are provided as the chiral
isomers or racemic mixtures. Both racemic and diastereomeric
mixtures, as well as the individual optical isomers isolated or
synthesized, substantially free of their enantiomeric or
diastereomeric partners, are all within the scope of the invention.
The racemic mixtures can be separated into their individual,
substantially optically pure isomers through well-known techniques
such as, for example, the separation of diastereomeric salts formed
with optically active adjuncts, e.g., acids or bases followed by
conversion back to the optically active substances. The desired
optical isomer can also be synthesized by means of stereospecific
reactions, beginning with the appropriate stereoisomer of the
desired starting material.
[0446] It is to be understood that for compounds disclosed herein
when a bond is drawn in a non-stereochemical manner (e.g., flat)
the atom to which the bond is attached includes all stereochemical
possibilities. It is also to be understood that when a bond is
drawn in a stereochemical manner (e.g., bold, bold-wedge, dashed or
dashed-wedge) the atom to which the stereochemical bond is attached
has the stereochemistry as shown unless otherwise noted.
Accordingly, in one embodiment, a compound disclosed herein is
greater than 50% a single enantiomer. In another embodiment, a
compound disclosed herein is at least 80% a single enantiomer. In
another embodiment, a compound disclosed herein is at least 90% a
single enantiomer. In another embodiment, a compound disclosed
herein is at least 98% a single enantiomer. In another embodiment,
a compound disclosed herein is at least 99% a single enantiomer. In
another embodiment, a compound disclosed herein is greater than 50%
a single diastereomer. In another embodiment, a compound disclosed
herein is at least 80% a single diastereomer. In another
embodiment, a compound disclosed herein is at least 90% a single
diastereomer. In another embodiment, a compound disclosed herein is
at least 98% a single diastereomer. In another embodiment, a
compound disclosed herein is at least 99% a single
diastereomer.
Tautomers
[0447] The compounds disclosed herein can also exist as tautomeric
isomers in certain cases. Although only one delocalized resonance
structure may be depicted, all such forms are contemplated within
the scope of the invention. For example, ene-amine tautomers can
exist for purine, pyrimidine, imidazole, guanidine, amidine, and
tetrazole systems and all their possible tautomeric forms are
within the scope of the invention.
Isotopes
[0448] It is understood by one skilled in the art that this
invention also includes any compound claimed that may be enriched
at any or all atoms above naturally occurring isotopic ratios with
one or more isotopes such as, but not limited to, deuterium
(.sup.2H or D). As a non-limiting example, a --CH.sub.3 group may
be replaced by --CD.sub.3.
[0449] Specific values listed below for radicals, substituents, and
ranges are for illustration only; they do not exclude other defined
values or other values within defined ranges for the radicals and
substituents.
Protecting Groups
[0450] In certain embodiments, protecting groups include prodrug
moieties and chemical protecting groups. Protecting groups may be
represented by the abbreviation "PG."
[0451] "Protecting group" ("PG") refers to a moiety of a compound
that masks or alters the properties of a functional group or the
properties of the compound as a whole. Chemical protecting groups
and strategies for protection/deprotection are well known in the
art. See e.g. Peter G. M. Wuts and Theodora W. Greene, Protective
Groups in Organic Synthesis, 4.sup.th edition; John Wiley &
Sons, Inc.: New Jersey, 2007. See also Kocienski, P. J. Protecting
Groups, 3.sup.rd edition; Georg Thieme Verlag Stuttgart: New York,
2005, in particular Chapter 1, Protecting Groups: An Overview,
pages 1-48, Chapter 2, Carbonyl Protecting Groups, pages 49-118,
Chapter 3, Diol Protecting Groups, pages 119-186, Chapter 4,
Hydroxyl Protecting Groups, pages 187-364, Chapter 5, Thiol
Protecting Groups, pages 365-392. Protecting groups are often
utilized to mask the reactivity of certain functional groups, to
assist in the efficiency of desired chemical reactions, e.g.,
making and breaking chemical bonds in an ordered and planned
fashion.
[0452] Protection of functional groups of a compound alters other
physical properties besides the reactivity of the protected
functional group, such as the polarity, lipophilicity
(hydrophobicity), and other properties which can be measured by
common analytical tools. Chemically protected intermediates may
themselves be biologically active or inactive.
[0453] In certain embodiments, protecting groups are optionally
employed to prevent side reactions with the protected group during
synthetic procedures. Selection of the appropriate groups to
protect, when to do so, and the nature of the chemical protecting
group "PG" is dependent upon the chemistry of the reaction to be
protected against (e.g., acidic, basic, oxidative, reductive or
other conditions) and the intended direction of the synthesis. PGs
do not need to be, and generally are not, the same if the compound
is substituted with multiple PG. In general, PG will be used to
protect functional groups such as carboxyl, hydroxyl, thio, or
amino groups and to thus prevent side reactions or to otherwise
facilitate the synthetic efficiency. The order of deprotection to
yield free deprotected groups is dependent upon the intended
direction of the synthesis and the reaction conditions to be
encountered, and may occur in any order as determined by the
artisan.
Salts and Hydrates
[0454] Examples of pharmaceutically acceptable salts of the
compounds disclosed herein include salts derived from an
appropriate base, such as an alkali metal (for example, sodium), an
alkaline earth metal (for example, magnesium), ammonium and
NX.sub.4 (wherein X is C.sub.1-C.sub.4 alkyl). Pharmaceutically
acceptable salts of a nitrogen atom or an amino group include for
example salts of organic carboxylic acids such as acetic, benzoic,
lactic, fumaric, tartaric, maleic, malonic, malic, isethionic,
lactobionic and succinic acids; organic sulfonic acids, such as
methanesulfonic, ethanesulfonic, benzenesulfonic and
p-toluenesulfonic acids; and inorganic acids, such as hydrochloric,
hydrobromic, sulfuric, phosphoric and sulfamic acids.
Pharmaceutically acceptable salts of a compound of a hydroxy group
include the anion of said compound in combination with a suitable
cation such as Na.sup.+ and NX.sub.4.sup.+ (wherein each X is
independently selected from H or a C.sub.1-C.sub.4 alkyl
group).
[0455] For therapeutic use, salts of active ingredients of the
compounds disclosed herein will typically be pharmaceutically
acceptable, i.e., they will be salts derived from a physiologically
acceptable acid or base. However, salts of acids or bases which are
not pharmaceutically acceptable may also find use, for example, in
the preparation or purification of a compound of Formula I, II, III
or IV, (such as any one of IVa-IVh) or a stereoisomer, or a mixture
of stereoisomers, or another compound disclosed herein. All salts,
whether or not derived from a physiologically acceptable acid or
base, are within the scope of the present invention.
[0456] Metal salts typically are prepared by reacting the metal
hydroxide with a compound disclosed herein. Examples of metal salts
which are prepared in this way are salts containing Li.sup.+,
Na.sup.+, and K.sup.+. A less soluble metal salt can be
precipitated from the solution of a more soluble salt by addition
of the suitable metal compound.
[0457] In addition, salts may be formed from acid addition of
certain organic and inorganic acids, e.g., HCl, HBr,
H.sub.2SO.sub.4, H.sub.3PO.sub.4 or organic sulfonic acids, to
basic centers, such as amines. Finally, it is to be understood that
the compositions herein comprise compounds disclosed herein in
their un-ionized, as well as zwitterionic form, and combinations
with stoichiometric amounts of water as in hydrates.
EMBODIMENTS
[0458] In certain embodiments, A is --C(O)--, 6-10 membered
arylene, or 5-6 membered heteroarylene group, wherein said arylene
or heteroarylene is optionally substituted with 1-4 halogens or
haloalkyl groups. In some embodiments, A is --C(O)--.
[0459] In certain embodiments, M is --O-- or a bond. In some
embodiments, M is --O--.
[0460] In certain embodiments, G is --CO.sub.2H or
--CONHSO.sub.2Z.sup.2. In some embodiments, G is
--CONHSO.sub.2Z.sup.2. In some embodiments, G is
--CONHSO.sub.2Z.sup.2 and Z.sup.2 is cyclopropyl optionally
substituted with methyl.
[0461] In certain embodiments, G is:
##STR00019##
[0462] In some embodiments, G is:
##STR00020##
[0463] In certain embodiments, Z.sup.2 is:
##STR00021##
[0464] In certain embodiments, Z.sup.2 is:
##STR00022##
[0465] In certain embodiments, Z.sup.2a is hydrogen, halogen or
methyl. In some embodiments, Z.sup.2a is:
##STR00023##
[0466] In other embodiments, Z.sup.2a is
##STR00024##
(i.e., hydrogen or methyl).
[0467] In other embodiments, Z.sup.2a is
##STR00025##
[0468] In still more embodiments, Z.sup.2a is (i.e., methyl).
##STR00026##
[0469] In certain embodiments, one of R.sup.3, R.sup.4, and R.sup.5
is Z.sup.1 and the other two are H. In some embodiments, R.sup.3,
R.sup.4 and R.sup.5 are each H.
[0470] In certain embodiments, X is --OC(O)--, --O--, or a direct
bond. In some embodiments, X is --O--.
[0471] In certain embodiments, X is --OC(O)--, --O--, or a direct
bond. In certain other embodiments, X is --O--.
[0472] In certain embodiments, {circle around (T)} is
C.sub.3-C.sub.6 carbocyclylene that is attached to L and to the
remainder of the compound of Formula I, II, III, or IV (such as
IVa-IVh) through two adjacent carbons, wherein said C.sub.3-C.sub.5
carbocyclylene is optionally substituted with C.sub.1-C.sub.4 alkyl
or C.sub.1-C.sub.3 haloalkyl.
[0473] In certain embodiments, {circle around (T)} is
C.sub.3-C.sub.6 carbocyclylene that is attached to L and to the
remainder of the compound of Formula I, II, III, or IV (such as any
one of IVa-IVh) through two adjacent carbons, wherein the
C.sub.3-C.sub.5 carbocyclylene is optionally substituted with
methyl, ethyl or trifluoromethyl. In some embodiments, {circle
around (T)} is C.sub.3-C.sub.6 carbocyclylene that is attached to L
and to the remainder of the compound of Formula I, II, III, or IV
(such as any one of IVa-IVh) through two adjacent carbons.
[0474] In certain embodiments, {circle around (T)} is
C.sub.3-C.sub.6 cycloalkyl that is attached to L and to the
remainder of the compound of Formula I, II, III, or IV (such as any
one of IVa-IVh) through two adjacent carbons, wherein the
C.sub.3-C.sub.5 cycloalkyl is optionally substituted with methyl,
ethyl or trifluoromethyl. In some embodiments, {circle around (T)}
is C.sub.3-C.sub.6 cycloalkyl that is attached to L and to the
remainder of the compound of Formula I, II, III, or IV (such as any
one of IVa-IVh) through two adjacent carbons.
[0475] In certain embodiments, {circle around (T)} is cyclopropyl
optionally substituted with methyl or trifluoromethyl.
[0476] In certain embodiments, {circle around (T)} is
C.sub.6-C.sub.8 bridged bicyclic carbocyclylene or C.sub.6-C.sub.8
bridged fused carbocyclylene that is attached to L and to the
remainder of the compound of Formula I, II, III, or IV (such as any
one of IVa-IVh) through two adjacent carbons.
[0477] In certain embodiments, {circle around (T)} is
C.sub.6-C.sub.8 bridged bicyclic cycloalkyl or C.sub.6-C.sub.8
bridged fused cycloalkyl that is attached to L and to the remainder
of the compound of Formula I, II, III, or IV (such as any one of
IVa-IVh) through two adjacent carbons.
[0478] In some embodiments, {circle around (T)} is T.sup.1,
T.sup.2, T.sup.3, T.sup.4, T.sup.5, T.sup.8, T.sup.9, T.sup.10,
T.sup.11, or T.sup.12. In certain embodiments, {circle around (T)}
is T.sup.1, T.sup.2, T.sup.3, T.sup.4, T.sup.9, T.sup.10, T.sup.11
or T.sup.14. In some embodiments, {circle around (T)} is T.sup.1,
T.sup.2, or T.sup.3, optionally substituted with 1-4 Z.sup.1 groups
which are the same or different.
[0479] In some embodiments, T.sup.1 is
##STR00027##
[0480] In some embodiments, T.sup.2 is
##STR00028##
[0481] In some embodiments, T.sup.3 is
##STR00029##
[0482] In some embodiments, T.sup.5 is
##STR00030##
[0483] In some embodiments, T.sup.8 is
##STR00031##
##STR00032##
[0484] In some embodiments, T.sup.9 is
##STR00033##
[0485] In some embodiments, T.sup.10 is
##STR00034##
[0486] In some embodiments, T.sup.11 is
##STR00035##
[0487] In some embodiments, T.sup.12 is
##STR00036##
[0488] In other embodiments, {circle around (T)} is
##STR00037##
[0489] In certain embodiments, T.sup.1 is:
##STR00038##
[0490] In certain embodiments, T.sup.2 is:
##STR00039##
[0491] In certain embodiments, T.sup.3 is:
##STR00040##
[0492] In certain embodiments, {circle around (T)} is T.sup.2,
which is optionally substituted with 1-4 Z.sup.1 groups, which are
the same or different.
[0493] In certain embodiments, T.sup.2 is:
##STR00041##
[0494] In certain embodiments, {circle around (T)} is
##STR00042##
[0495] In other embodiments, {circle around (T)} is
##STR00043##
[0496] In other embodiments, {circle around (T)} is
##STR00044##
[0497] In other embodiments, {circle around (T)} is
##STR00045##
[0498] In certain embodiments, T.sup.2 is:
##STR00046##
[0499] In certain embodiments, T.sup.2 is:
##STR00047##
[0500] In certain embodiments, T.sup.2 is:
##STR00048##
(a stereoisomer of bicyclo[3.1.0]hexanylene).
[0501] In certain embodiments, J is J.sup.1, J.sup.4, J.sup.5 or
J.sup.8. In other embodiments, J is J.sup.4. In certain
embodiments, J is J.sup.5.
[0502] In further embodiments, J.sup.4 is
##STR00049##
[0503] In other embodiments, J is
##STR00050##
[0504] In other embodiments, J is
##STR00051##
[0505] In other embodiments, J.sup.5 is
##STR00052##
[0506] In certain embodiments, J is C.sub.1-C.sub.3 alkyl. In
certain embodiments, J is methyl or ethyl. In further other
embodiments, J is --CH.sub.2--CH.sub.3.
[0507] In some embodiments, L is L.sup.1, L.sup.2, L.sup.3,
L.sup.4, L.sup.5, L.sup.6, L.sup.7, L.sup.8 or L.sup.9. In one
embodiment, L is L.sup.1, L.sup.2, L.sup.3, L.sup.4, L.sup.5 or
L.sup.6. In certain embodiments, L is L.sup.1 or L.sup.2.
[0508] In some embodiments, L is C.sub.3-C.sub.6 alkylene,
substituted with 1-4 halogens.
[0509] In some embodiments, L is C.sub.5 alkylene, substituted with
two halogens. In some embodiments, the halogens of L are each
fluoro.
[0510] In certain embodiments, L is:
##STR00053##
[0511] In certain other embodiments, L is
##STR00054##
[0512] In some embodiments, L.sup.1 is:
##STR00055##
[0513] In some embodiments, L.sup.2 is:
##STR00056##
[0514] In some embodiments, L.sup.3 is
##STR00057##
[0515] In some embodiments, L.sup.4 is
##STR00058##
[0516] In some embodiments, L.sup.5 is
##STR00059##
[0517] In some embodiments, L.sup.6 is
##STR00060##
[0518] In some embodiments, L.sup.7 is
##STR00061##
[0519] In some embodiments, L.sup.8 is
##STR00062##
[0520] In some embodiments, L.sup.9 is
##STR00063##
[0521] In other embodiments, L is
##STR00064##
[0522] In further embodiments, L is
##STR00065##
[0523] In further embodiments, L is
##STR00066##
[0524] In still more embodiments, L is
##STR00067##
[0525] In other embodiments, L is
##STR00068##
[0526] In some embodiments, Q is Q.sup.1, Q.sup.2, Q.sup.3,
Q.sup.4, Q.sup.5 or Q.sup.7
[0527] In some embodiments, Q.sup.1 is
##STR00069##
[0528] In some embodiments, Q.sup.2 is
##STR00070##
[0529] In some embodiments, Q.sup.3 is
##STR00071##
[0530] In some embodiments, Q.sup.4 is
##STR00072##
[0531] In some embodiments, Q.sup.5 is or
##STR00073##
[0532] In some embodiments, Q.sup.7 is
##STR00074##
[0533] In other embodiments, Q is
##STR00075##
[0534] In certain embodiments, Q is Q.sup.1. In certain other
embodiments, Q is C.sub.1-C.sub.4 alkyl or C.sub.3-6 carbocyclyl.
In further embodiments, Q is
##STR00076##
(i.e., t-butyl). In some embodiments, Q is t-butyl or
C.sub.5-C.sub.6 cycloalkyl.
[0535] In certain embodiments, E is E.sup.1, E.sup.2, E.sup.3, or
E.sup.4. In certain embodiments, E is E.sup.3.
[0536] In certain embodiments, E is C.sub.1-C.sub.3 alkyl
optionally substituted with 1-3 halogen atoms. In certain
embodiments, E is difluoromethyl.
[0537] In certain other embodiments, E is
##STR00077##
[0538] In some embodiments, E is
##STR00078##
[0539] In other embodiments, E is
##STR00079##
[0540] In other embodiments, E is
##STR00080##
[0541] In other embodiments, E is
##STR00081##
[0542] In certain embodiments, {circle around (U)} is bicyclic
heteroaryl, optionally substituted with 1-4 W groups which are the
same or different.
[0543] In certain other embodiments, {circle around (U)} is
##STR00082##
optionally substituted with 1-4 W groups, which are the same or
different.
[0544] In certain embodiments, {circle around (U)} is substituted
with one W group.
[0545] In some embodiments, {circle around (U)} is U.sup.1,
U.sup.3, U.sup.4, U.sup.5 or U.sup.6, wherein each U.sup.1,
U.sup.3, U.sup.4, U.sup.5 or U.sup.6 is optionally substituted with
1-3 W at any substitutable position, and each W is independently
W.sup.1, W.sup.2, W.sup.3, W.sup.4, W.sup.5, W.sup.6 or
W.sup.7.
[0546] In certain embodiments, W.sup.1 is oxo, halogen, --OR.sup.6,
C.sub.1-C.sub.6 alkyl, --CN, --CF.sub.3, --SR.sup.6,
--C(O).sub.2R.sup.6, --C(O)N(R.sup.6).sub.2, --C(O)R.sup.6,
--N(R.sup.6)C(O)R.sup.6, --SO.sub.2(C.sub.1-C.sub.6 alkyl),
--S(O)(C.sub.1-C.sub.6 alkyl), C.sub.3-C.sub.8 carbocyclyl,
C.sub.3-C.sub.8 cycloalkoxy, C.sub.1-C.sub.6 haloalkyl,
--N(R.sup.6).sub.2, --NR.sup.6(C.sub.1-C.sub.6
alkyl)O(C.sub.1-C.sub.6 alkyl), halo(C.sub.1-C.sub.6 alkoxy),
--NR.sup.6SO.sub.2R.sup.6, --SO.sub.2N(R.sup.6).sub.2,
--NHCOOR.sup.6, --NHCONHR.sup.6, C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl, 4-10 membered heterocyclyl or --O(4-10
membered heterocyclyl), wherein said W alkyl, carbocyclyl,
cycloalkoxy, haloalkyl, haloalkoxy, aryl, heteroaryl, or
heterocyclyl is optionally substituted with 1-4 Z.sup.1c
groups.
[0547] In certain embodiments, each R.sup.6 is independently H,
C.sub.6-C.sub.10 aryl or C.sub.1-C.sub.6 alkyl, wherein said aryl
or alkyl is optionally substituted with 1 to 4 substituents
independently selected from halogen atoms, C.sub.1-C.sub.6 alkyl,
C.sub.6-C.sub.10 aryl, C.sub.3-C.sub.8 carbocyclyl, 5-14 membered
heteroaryl, 4-10 membered heterocyclyl, halo(C.sub.1-C.sub.6
alkoxy), --OH, --O(C.sub.1-C.sub.6 alkyl), --SH,
--S(C.sub.1-C.sub.6 alkyl), --NH.sub.2, --NH(C.sub.1-C.sub.6
alkyl), --N(C.sub.1-C.sub.6 alkyl).sub.2, --C(O)(C.sub.1-C.sub.6
alkyl), --SO.sub.2N(C.sub.1-C.sub.6 alkyl).sub.2,
--NHCOO(C.sub.1-C.sub.6 alkyl), --NHCO(C.sub.1-C.sub.6 alkyl),
--NHCONH(C.sub.1-C.sub.6 alkyl), --CO.sub.2(C.sub.1-C.sub.6 alkyl),
or --C(O)N(C.sub.1-C.sub.6 alkyl).sub.2.
[0548] In certain embodiments, W.sup.2 is C.sub.1-C.sub.6 alkoxy
substituted with a 5-14 membered heteroaryl or C.sub.6-C.sub.10
aryl; wherein said heteroaryl or aryl is substituted with 1-4
Z.sup.1c groups.
[0549] In certain embodiments, W.sup.3 is a C.sub.2-C.sub.8 alkynyl
group substituted with a C.sub.6-C.sub.10 aryl, C.sub.3-C.sub.8
carbocyclyl, C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.6 haloalkyl, 4-10
membered heterocyclyl, or 5-14 membered heteroaryl group; wherein
said aryl, carbocyclyl, alkyl, haloalkyl, heterocyclyl, or
heteroaryl group is optionally substituted with 1-4 Z.sup.1c
groups.
[0550] In some embodiments, W.sup.4 is --SF.sub.5.
[0551] In some embodiments, W.sup.5 is --O(C.sub.2-C.sub.6
alkyl)OR.sup.22 wherein R.sup.22 is a C.sub.6-C.sub.10 aryl, 5-14
membered heteroaryl or 4-10 membered heterocyclyl group that is
optionally substituted with 1-4 Z.sup.1c groups.
[0552] In certain embodiments, W is hydrogen,
--O(C.sub.1-C.sub.3)alkyl, halogen or cyano.
[0553] In certain embodiments, W is methoxy.
[0554] In some embodiments, U.sup.1 is
##STR00083##
wherein each U.sup.1 is optionally substituted with 1-2 Z.sup.1
groups.
[0555] In some embodiments, U.sup.3 is
##STR00084##
wherein each U.sup.3 is optionally substituted with 1-2 Z.sup.1
groups.
[0556] In some embodiments, U.sup.4 is
##STR00085##
wherein each U.sup.4 is optionally substituted with 1-2 Z.sup.1
groups.
[0557] In some embodiments, U.sup.5 is
##STR00086## ##STR00087##
wherein each U.sup.5 is optionally substituted with 1-2 Z.sup.1
groups.
[0558] In some embodiments, U.sup.6 is
##STR00088##
wherein each U.sup.6 is optionally substituted with 1-2 Z.sup.1
groups.
[0559] In some embodiments, U.sup.7 is
##STR00089##
wherein each U.sup.7 is optionally substituted with 1-2 Z.sup.1
groups.
[0560] In other embodiments, {circle around (T)} is optionally
substituted with one or two W at any substitutable position, and
each W is independently W.sup.1, W.sup.2, W.sup.3, W.sup.4,
W.sup.5, W.sup.6 or W.sup.7 wherein (-) is
##STR00090##
[0561] In other embodiments, {circle around (U)} is optionally
substituted with one W at any substitutable position, and each W is
independently W.sup.1, W.sup.2, W.sup.3, W.sup.4, W.sup.5, W.sup.6
or W.sup.7 wherein {circle around (U)} is
##STR00091##
[0562] In other embodiments, each W is independently W.sup.1,
W.sup.2, W.sup.3, W.sup.4, W.sup.5, W.sup.6 or W.sup.7.
[0563] In some embodiments, W.sup.1 is
##STR00092##
[0564] In some embodiments, W.sup.2 is
##STR00093##
[0565] In some embodiments, W.sup.3 is
##STR00094##
[0566] In some embodiments, W.sup.5 is
##STR00095##
[0567] In some embodiments, W.sup.6 is
##STR00096##
[0568] In some embodiments, W.sup.7 is
##STR00097##
[0569] In other embodiments, W is
##STR00098##
[0570] In certain embodiments, each W is independently halogen or
C.sub.1-C.sub.4 alkoxy.
[0571] In a specific embodiment, J is methyl or ethyl; E is
substituted with 1-2 halogen atoms; L is substituted with
1-2-halogen atoms, and T is unsubstituted.
[0572] In a further embodiment, J is methyl or ethyl; E is
C.sub.1-C.sub.3 haloalkyl; L is C.sub.5-alkyl or
C.sub.5-alkenyl.
[0573] In another embodiment, a compound of Formula (IV):
##STR00099##
or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof, is provided, wherein:
[0574] J is C.sub.1-C.sub.4 alkyl or C.sub.3-C.sub.6 carbocyclyl,
wherein C.sub.1-C.sub.4 alkyl or C.sub.3-C.sub.6 carbocyclyl is
optionally substituted with halogen, --OH, aryl or cyano; [0575]
{circle around (T)} is C.sub.3-C.sub.5 carbocyclylene that is
attached to L and to the remainder of the compound through two
adjacent carbons, wherein said C.sub.3-C.sub.5 carbocyclylene is
optionally substituted with C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.3
haloalkyl, halogen, --OH, or cyano, or {circle around (T)} is
C.sub.5-C.sub.8 bicyclic carbocyclylene that is attached to L and
to the remainder of the compound through two adjacent carbons, or
{circle around (T)} is C.sub.3-C.sub.6 carbocyclylene that is
attached to L and to the remainder of the compound of Formula IV
through two adjacent carbons, wherein said C.sub.3-C.sub.6
carbocyclene is optionally substituted with C.sub.1-C.sub.4 alkyl
or C.sub.1-C.sub.3 haloalkyl; [0576] L is C.sub.3-C.sub.6 alkylene,
C.sub.3-C.sub.6 alkenylene or --(CH.sub.2).sub.3-cyclopropylene-,
optionally substituted with 1-4 halogen, --OH, or cyano; [0577] Q
is C.sub.2-C.sub.4 alkyl or C.sub.3-C.sub.6 carbocyclyl optionally
substituted with C.sub.1-C.sub.3 alkyl, halogen, --OH, or cyano;
[0578] E is C.sub.1-C.sub.3 alkyl or C.sub.2-C.sub.3 alkenyl,
optionally substituted with C.sub.1-C.sub.3 alkyl, halogen, --OH,
or cyano; [0579] W is H, --OH, --O(C.sub.1-C.sub.3)alkyl,
--O(C.sub.1-C.sub.3)haloalkyl, halogen or cyano; and [0580]
Z.sup.2a is H or C.sub.1-C.sub.3 alkyl, halogen, --OH, or
cyano.
[0581] In a further embodiment of Formula (IV), J is
C.sub.1-C.sub.3 alkyl.
[0582] In a further embodiment of Formula (IV), J is methyl or
ethyl.
[0583] In a further embodiment of Formula (IV), {circle around (T)}
is C.sub.3-C.sub.6 carbocyclylene that is attached to L and to the
remainder of the compound of Formula IV through two adjacent
carbons, wherein said C.sub.3-C.sub.6 carbocyclene is optionally
substituted with C.sub.1-C.sub.4 alkyl or C.sub.1-C.sub.3
haloalkyl.
[0584] In a further embodiment of Formula (IV), {circle around (T)}
is C.sub.3-C.sub.6 carbocyclylene that is attached to L and to the
remainder of the compound of Formula IV through two adjacent
carbons, wherein the C.sub.3-C.sub.6 carbocyclene is optionally
substituted with methyl, ethyl or trifluoromethyl.
[0585] In a further embodiment of Formula (IV), {circle around (T)}
is cyclopropylene.
[0586] In a further embodiment of Formula (IV), {circle around (T)}
is C.sub.6-C.sub.8 bridged bicyclic carbocyclylene or
C.sub.6-C.sub.8 fused bicyclic carbocyclylene that is attached to L
and to the remainder of the compound of Formula IV through two
adjacent carbons.
[0587] In a further embodiment of Formula (IV), L is
C.sub.3-C.sub.6 alkylene, substituted with 1-4 halogens. In another
embodiment of Formula (IV), L is C.sub.5 alkylene, substituted with
two halogens. In some embodiments, the halogens are each
fluoro.
[0588] In a further embodiment of Formula (IV), L is
C.sub.3-C.sub.6 alkylene.
[0589] In a further embodiment of Formula (IV), L is C.sub.5
alkylene.
[0590] In a further embodiment of Formula (IV), Q is t-butyl or
C.sub.5-C.sub.6 carbocyclyl.
[0591] In a further embodiment of Formula (IV), Q is t-butyl.
[0592] In a further embodiment of Formula (IV), E is
C.sub.1-C.sub.3 alkyl optionally substituted with 1-3 halogen
atoms.
[0593] In a further embodiment of Formula (IV), E is
difluoromethyl.
[0594] In a further embodiment of Formula (IV), W is hydrogen,
--O(C.sub.1-C.sub.3)alkyl, halogen or cyano.
[0595] In a further embodiment of Formula (IV), W is methoxy.
[0596] In a further embodiment of Formula (IV), Z.sup.2a is
hydrogen or methyl.
[0597] In a further embodiment of Formula (IV), Z.sup.2a is
methyl.
[0598] There is further provided a compound selected from the group
consisting of:
##STR00100## ##STR00101## ##STR00102## ##STR00103## ##STR00104##
##STR00105## ##STR00106## ##STR00107## ##STR00108## ##STR00109##
##STR00110## ##STR00111## ##STR00112## ##STR00113## ##STR00114##
##STR00115## ##STR00116## ##STR00117## ##STR00118## ##STR00119##
##STR00120## ##STR00121## ##STR00122## ##STR00123##
##STR00124##
[0599] In one embodiment, a compound of Formula IVa, or a
pharmaceutically acceptable salt thereof, is provided:
##STR00125##
[0600] In one embodiment, a compound of Formula IVb: or a
pharmaceutically acceptable salt thereof, is provided:
##STR00126##
[0601] In one embodiment, a compound of Formula IVc, or a
pharmaceutically acceptable salt thereof, is provided:
##STR00127##
[0602] In one embodiment, a compound of Formula IVd, or a
pharmaceutically acceptable salt thereof, is provided:
##STR00128##
[0603] In one embodiment, a compound of Formula IVe, or a
pharmaceutically acceptable salt thereof, is provided:
##STR00129##
[0604] In one embodiment, a compound of Formula IVf, or a
pharmaceutically acceptable salt thereof, is provided:
##STR00130##
[0605] In one embodiment, a compound of Formula IVg, or a
pharmaceutically acceptable salt thereof, is provided:
##STR00131##
[0606] In one embodiment, a compound of Formula IVh, or a
pharmaceutically acceptable salt thereof, is provided:
##STR00132##
[0607] In one embodiment, a compound of any one of Formula IVa,
IVb, IVc, IVd, IVe, IVf, IVg, or IVh, or a stereoisomer, or a
mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, is provided.
Methods of Treatment
[0608] One embodiment provides a method for treating a Flaviviridae
viral infection (e.g., an HCV viral infection) in a patient in need
thereof (e.g., a mammal such as a human). The method includes
administering a compound of Formula I, II III or IV (such as any
one of IVa-IVh), or a stereoisomer, or a mixture of stereoisomers,
or a pharmaceutically acceptable salt thereof, to the patient.
[0609] One embodiment provides a method for inhibiting the
proliferation of the HCV virus, treating HCV infection or delaying
the onset of HCV symptoms in a patient in need thereof (e.g., a
mammal such as a human). The method includes administering a
compound of Formula I, II, III or IV (such as any one of IVa-IVh),
or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof, to the patient.
[0610] One embodiment provides a compound of Formula I, II, III or
IV (such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof for
use in medical therapy (e.g., for use in treating a Flaviviridae
viral infection (e.g., an HCV viral infection) or the proliferation
of the HCV virus or delaying the onset of HCV symptoms in a patient
(e.g., a mammal such as a human)).
[0611] One embodiment provides a compound of Formula I, II, III, or
IV (such as any one of IVa-IVh) or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof for
use in the manufacture of a medicament for treating a Flaviviridae
viral infection (e.g., an HCV viral infection) or the proliferation
of the HCV virus or delaying the onset of HCV symptoms in a patient
in need thereof (e.g., mammal such as a human).
[0612] One embodiment provides a compound of Formula I, II, III, or
IV (such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, for
use in the prophylactic or therapeutic treatment of the
proliferation of a Flaviviridae virus, an HCV virus or for use in
the therapeutic treatment of delaying the onset of HCV
symptoms.
[0613] One embodiment provides a compound of Formula I, II, III or
IV (such as any one of IVa-IVh) or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, for
use in the prophylactic or therapeutic treatment of a Flaviviridae
virus infection (e.g., an HCV virus infection).
[0614] One embodiment provides the use of a compound of Formula I,
II, III or IV (such as any one of IVa-IVh), or a stereoisomer, or a
mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, for the manufacture of a medicament for a Flaviviridae
virus infection (e.g., an HCV virus infection) in a mammal (e.g., a
human).
[0615] In certain embodiments, a method of treating chronic
hepatitis C infection is provided. The method includes
administering to a patient in need thereof, a compound of Formula
I, II III or IV (such as any one of IVa-IVh), or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof, to the patient.
[0616] In certain embodiments, a method of treating hepatitis C
infection in treatment-naive patients is provided. The method
includes administering to a treatment-naive patient, a compound of
Formula I, II III or IV (such as any one of IVa-IVh), or a
stereoisomer, or a mixture of stereoisomers, or a pharmaceutically
acceptable salt thereof.
[0617] In certain embodiments, a method of treating hepatitis C
infection in treatment-experienced patients is provided. The method
includes administering to a treatment-experienced patient, a
compound of Formula I, II III or IV (such as any one of IVa-IVh),
or a stereoisomer, or a mixture of stereoisomers, or a
pharmaceutically acceptable salt thereof.
[0618] In certain embodiments, a method of treating hepatitis C
infection in an interferon ineligible or an interferon intolerant
patient is provided. The method includes administering, a compound
of Formula I, II III or IV (such as any one of IVa-IVh), or a
stereoisomer, or a mixture of stereoisomers, or a pharmaceutically
acceptable salt thereof, to the patient.
[0619] In certain embodiments, the methods of treatment described
herein include administering the compound of Formula I, II III or
IV (such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient for a fixed period of duration. In some embodiments,
the fixed period of duration is 4 weeks, 6 weeks, 8 weeks, 10 weeks
or 12 weeks. In other embodiments, the fixed period of duration is
not more than 12 weeks.
[0620] In some embodiments, the compound is administered for about
12 weeks. In further embodiments, the compound is administered for
about 12 weeks or less, for about 10 weeks or less, for about 8
weeks or less, for about 6 weeks or less, or for about 4 weeks or
less.
[0621] The compound may be administered once daily, twice daily,
once every other day, two times a week, three times a week, four
times a week, or five times a week.
[0622] In certain embodiments, the methods of treatment described
herein includes administering a compound of Formula I, II III or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to is
infected with HCV genotype (GT) 1, 2, 3, 4, 5, or 6 (i.e., a method
for treating a GT 1, 2, 3, 4, 5, or 6 HCV infection).
[0623] One embodiment provides a method for treating an HCV
infection in a patient in need thereof (e.g., a mammal such as a
human), wherein the patient is infected with HCV genotype 1. The
method includes administering a compound of Formula I, II III or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient.
[0624] One embodiment provides a method for treating an HCV
infection in a patient in need thereof (e.g., a mammal such as a
human), wherein the patient is infected with HCV genotype 2. The
method includes administering a compound of Formula I, II III or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient.
[0625] One embodiment provides a method for treating an HCV
infection in a patient in need thereof (e.g., a mammal such as a
human), wherein the patient is infected with HCV genotype 3. The
method includes administering a compound of Formula I, II III or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient.
[0626] One embodiment provides a method for treating an HCV
infection in a patient in need thereof (e.g., a mammal such as a
human), wherein the patient is infected with HCV genotype 4. The
method includes administering a compound of Formula I, II III or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient.
[0627] One embodiment provides a method for treating an HCV
infection in a patient in need thereof (e.g., a mammal such as a
human), wherein the patient is infected with HCV genotype 5. The
method includes administering a compound of Formula I, II III or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient.
[0628] One embodiment provides a method for treating an HCV
infection in a patient in need thereof (e.g., a mammal such as a
human), wherein the patient is infected with HCV genotype 6. The
method includes administering a compound of Formula I, II III or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient.
[0629] In the methods of treatment described herein, the
administering step includes administering a therapeutically
effective amount of a compound of Formula I, II III or IV (such as
any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof, to
the patient in need of treatment.
[0630] In certain embodiments, methods of inhibiting the activity
of HCV are provided. Such methods include the step of treating a
sample suspected of containing HCV with a compound or composition
disclosed herein.
[0631] In one embodiment, compounds disclosed herein act as
inhibitors of HCV, as intermediates for such inhibitors or have
other utilities as described below.
[0632] In certain embodiments, compounds binding in the liver may
bind with varying degrees of reversibility.
[0633] In one embodiment, a method for treating HCV includes adding
a compound disclosed herein to the sample. The addition step
comprises any method of administration as described above.
[0634] If desired, the activity of HCV after application of the
compound can be observed by any method including direct and
indirect methods of detecting HCV activity. Quantitative,
qualitative, and semiquantitative methods of determining HCV
activity are all contemplated. Typically one of the screening
methods described above are applied, however, any other method such
as observation of the physiological properties of a living organism
are also applicable.
[0635] Many organisms contain HCV. The compounds of this invention
are useful in the treatment or prophylaxis of conditions associated
with HCV activation in animals or in humans.
Pharmaceutical Formulations
[0636] "Pharmaceutically-acceptable" means suitable for use in
pharmaceutical preparations, generally considered as safe for such
use, officially approved by a regulatory agency of a national or
state government for such use, or being listed in the U. S.
Pharmacopoeia or other generally recognized pharmacopoeia for use
in animals, and more particularly in humans.
[0637] "Pharmaceutically-acceptable carrier" refers to a diluent,
adjuvant, excipient, or carrier, or other ingredient which is
pharmaceutically-acceptable and with which a compound of the
invention is administered.
[0638] The compounds of this invention are formulated with
conventional carriers (e.g., inactive ingredient or excipient
material), which will be selected in accord with ordinary practice.
Tablets will contain excipients including glidants, fillers,
binders and the like. Aqueous formulations are prepared in sterile
form, and when intended for delivery by other than oral
administration generally will be isotonic. All formulations will
optionally contain excipients such as those set forth in the
Handbook of Pharmaceutical Excipients (1986). Excipients include
ascorbic acid and other antioxidants, chelating agents such as
EDTA, carbohydrates such as dextrin, hydroxyalkylcellulose,
hydroxyalkylmethylcellulose, stearic acid and the like. One
embodiment provides the formulation as a solid dosage form
including a solid oral dosage form. The pH of the formulations
ranges from about 3 to about 11, but is ordinarily about 7 to
10.
[0639] While it is possible for the active ingredients to be
administered alone it may be preferable to present them as
pharmaceutical formulations (compositions). The formulations, both
for veterinary and for human use, of the invention comprise at
least one active ingredient, as above defined, together with one or
more acceptable carriers therefor and optionally other therapeutic
ingredients. The carrier(s) must be "acceptable" in the sense of
being compatible with the other ingredients of the formulation and
physiologically innocuous to the recipient thereof.
[0640] The formulations include those suitable for the foregoing
administration routes. The formulations may conveniently be
presented in unit dosage form and may be prepared by any of the
methods well known in the art of pharmacy. Techniques and
formulations generally are found in Remington's Pharmaceutical
Sciences (Mack Publishing Co., Easton, Pa.). Such methods include
the step of bringing into association the active ingredient with
inactive ingredients (e.g., a carrier, pharmaceutical excipient,
etc.) which constitutes one or more accessory ingredients. In
general the formulations are prepared by uniformly and intimately
bringing into association the active ingredient with liquid
carriers or finely divided solid carriers or both, and then, if
necessary, shaping the product.
[0641] In certain embodiments, formulations suitable for oral
administration are presented as discrete units such as capsules,
cachets or tablets each containing a predetermined amount of the
active ingredient.
[0642] In certain embodiments, the pharmaceutical formulations
include one or more compounds of the invention together with one or
more pharmaceutically acceptable carriers or excipients and
optionally other therapeutic agents. Pharmaceutical formulations
containing the active ingredient may be in any form suitable for
the intended method of administration. When used for oral use for
example, tablets, troches, lozenges, aqueous or oil suspensions,
dispersible powders or granules, emulsions, hard or soft capsules,
syrups or elixirs may be prepared. Compositions intended for oral
use may be prepared according to any method known to the art for
the manufacture of pharmaceutical compositions and such
compositions may contain one or more agents including sweetening
agents, flavoring agents, coloring agents and preserving agents, in
order to provide a palatable preparation. Tablets containing the
active ingredient in admixture with non-toxic pharmaceutically
acceptable excipient which are suitable for manufacture of tablets
are acceptable. These excipients may be, for example, inert
diluents, such as calcium or sodium carbonate, lactose, lactose
monohydrate, croscarmellose sodium, povidone, calcium or sodium
phosphate; granulating and disintegrating agents, such as maize
starch, or alginic acid; binding agents, such as cellulose,
microcrystalline cellulose, starch, gelatin or acacia; and
lubricating agents, such as magnesium stearate, stearic acid or
talc. Tablets may be uncoated or may be coated by known techniques
including microencapsulation to delay disintegration and adsorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a time delay material
such as glyceryl monostearate or glyceryl distearate alone or with
a wax may be employed.
[0643] The amount of active ingredient that is combined with the
inactive ingredients to produce a dosage form will vary depending
upon the host treated and the particular mode of administration.
For example, in some embodiments, a dosage form for oral
administration to humans contains approximately 1 to 1000 mg of
active material formulated with an appropriate and convenient
amount of carrier material (e.g., inactive ingredient or excipient
material). In certain embodiments, the carrier material varies from
about 5 to about 95% of the total compositions (weight: weight). In
some embodiments, the pharmaceutical compositions described herein
contain about 1 to 800 mg, 1 to 600 mg, 1 to 400 mg, 1 to 200 mg, 1
to 100 mg or 1 to 50 mg of the compound of Formula I, II, 111 or IV
(such as any one of IVa-IVh), or a stereoisomer, or a mixture of
stereoisomers, or a pharmaceutically acceptable salt thereof. In
some embodiments, the pharmaceutical compositions described herein
contain not more than about 400 mg of the compound of Formula I,
II, III or IV (such as any one of IVa-IVh), or a stereoisomer, or a
mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof. In some embodiments, the pharmaceutical compositions
described herein contain about 100 mg of the compound of Formula I,
II, III, or IV (such as any one of IVa-IVh), or a stereoisomer, or
a mixture of stereoisomers, or a pharmaceutically acceptable salt
thereof.
[0644] It should be understood that in addition to the ingredients
particularly mentioned above the formulations disclosed herein may
include other agents conventional in the art having regard to the
type of formulation in question, for example those suitable for
oral administration may include flavoring agents.
[0645] Veterinary compositions comprising at least one active
ingredient as above defined together with a veterinary carrier are
further provided.
[0646] Veterinary carriers are materials useful for the purpose of
administering the composition and may be solid, liquid or gaseous
materials which are otherwise inert or acceptable in the veterinary
art and are compatible with the active ingredient. These veterinary
compositions may be administered orally, parenterally or by any
other desired route.
[0647] Effective dose of active ingredient depends at least on the
nature of the condition being treated, toxicity, whether the
compound is being used prophylactically (lower doses), the method
of delivery, and the pharmaceutical formulation, and will be
determined by the clinician using conventional dose escalation
studies.
Routes of Administration
[0648] One or more compounds of Formulas I, II, III, or IV (such as
any one of IVa-IVh) (herein referred to as the active ingredients),
or a pharmaceutically acceptable salt thereof, are administered by
any route appropriate to the condition to be treated. Suitable
routes include oral, rectal, nasal, topical (including buccal and
sublingual), vaginal and parenteral (including subcutaneous,
intramuscular, intravenous, intradermal, intrathecal and epidural),
and the like. It will be appreciated that the preferred route may
vary with for example the condition of the recipient. An advantage
of the compounds of this invention is that they are orally
bioavailable and can be dosed orally. Accordingly, in one
embodiment, the pharmaceutical compositions described herein are
oral dosage forms. In certain embodiments, the pharmaceutical
compositions described herein are oral solid dosage forms.
[0649] One skilled in the art will recognize that substituents and
other moieties of the compounds of the generic formula herein
should be selected in order to provide a compound which is
sufficiently stable to provide a pharmaceutically useful compound
which can be formulated into an acceptably stable pharmaceutical
composition. Compounds which have such stability are contemplated
as falling within the scope of the present invention. It should be
understood by one skilled in the art that any combination of the
definitions and substituents described above should not result in
an inoperable species or compound.
Combination Therapy
[0650] In yet another embodiment, the present application discloses
pharmaceutical compositions comprising a compound of Formulas I,
II, III, or IV (such as any one of IVa-IVh), or a pharmaceutically
acceptable salt thereof, in combination with at least one
additional therapeutic agent (i.e., active ingredient), and a
pharmaceutically acceptable carrier or excipient. In certain
embodiments, additional therapeutic agents include additional
antiviral agents.
[0651] The additional therapeutic agent used in combination with
the compounds described herein includes, without limitation, any
agent having a therapeutic effect when used in combination with the
compound of the present invention. Such combinations are selected
based on the condition to be treated, cross-reactivities of
ingredients and pharmaco-properties of the combination. For
example, in certain embodiments, the therapeutic agent used in
combination with the compounds of Formulas I, II, III, or IV (such
as any one of IVa-IVh) include, without limitation, one of more of
the following: interferons, ribavirin analogs, NS3 protease
inhibitors, NS5a inhibitors, NS5b inhibitors, alpha-glucosidase 1
inhibitors, hepatoprotectants, non-nucleoside inhibitors of HCV,
nucleoside analogues, and other drugs for treating HCV infection.
In some embodiments, the additional therapeutic agents include,
without limitation, NS3 protease inhibitors, NS5a inhibitors,
and/or NS5b inhibitors. In some embodiments, a pharmaceutical
composition including a compound of Formulas I, II, III, or IV
(such as any one of IVa-IVh), or a pharmaceutically acceptable salt
thereof and one or more of an NS3 protease inhibitor, an NS5a
inhibitor, and/or an NS5b inhibitor is provided. In some
embodiments, a pharmaceutical composition including a compound of
Formulas I, II, III, or IV (such as any one of IVa-IVh), or a
pharmaceutically acceptable salt thereof and one or more of an NS5a
inhibitor and/or an NS5b inhibitor is provided. In certain
embodiments, pharmaceutical compositions is provided which includes
a compound of Formulas I, II, III, or IV (such as any one of
IVa-IVh) and one or more additional antiviral agents, wherein the
additional antiviral agent is not an interferon, ribavirin, or a
ribavirin analogue. In further embodiments, pharmaceutical
compositions is provided which includes a compound of Formulas I,
II, III, or IV (such as any one of IVa-IVh), or a stereoisomer, or
a mixture of stereoisomers, and one or more additional antiviral
agents, wherein the additional antiviral agent is not ribavirin or
a ribavirin analogue.
[0652] In certain embodiments, the compounds disclosed herein are
combined with one or more other active ingredients (e.g., one or
more additional antiviral agents) in a unitary dosage form for
simultaneous or sequential administration to a patient. The
combination therapy may be administered as a simultaneous or
sequential regimen. When administered sequentially, the combination
is administered in two or more administrations. In certain
embodiments, the active ingredients are: (1) co-formulated and
administered or delivered simultaneously in a combined
pharmaceutical composition; (2) delivered by alternation or in
parallel as separate pharmaceutical composition; or (3) by some
other regimen. When delivered in alternation therapy, the active
ingredients are administered or delivered sequentially, e.g., in
separate tablets, pills or capsules, or by different injections in
separate syringes. In general, during alternation therapy, an
effective dosage of each active ingredient is administered
sequentially, i.e. serially, whereas in combination therapy,
effective dosages of two or more active ingredients are
administered together.
[0653] Exemplary inferferons include, without limitation, pegylated
rIFN-alpha 2b (PEG-Intron), pegylated rIFN-alpha 2a (Pegasys),
rIFN-alpha 2b (Intron A), rIFN-alpha 2a (Roferon-A), interferon
alpha (MOR-22, OPC-18, Alfaferone, Alfanative, Multiferon,
subalin), interferon alfacon-1 (Infergen), interferon alpha-n1
(Wellferon), interferon alpha-n3 (Alferon), interferon-beta
(Avonex, DL-8234), interferon-omega (omega DUROS, Biomed 510),
albinterferon alpha-2b (Albuferon), IFN alpha XL, BLX-883
(Locteron), DA-3021, glycosylated interferon alpha-2b (AVI-005),
PEG-Infergen, PEGylated interferon lambda (PEGylated IL-29), or
belerofon, IFN alpha-2b XL, rIFN-alpha 2a, consensus IFN alpha,
infergen, rebif, pegylated IFN-beta, oral interferon alpha, feron,
reaferon, intermax alpha, r-IFN-beta, and infergen+actimmune.
[0654] Exemplary ribavarin analogs include, without limitation,
ribavirin (Rebetol, Copegus), levovirin VX-497, and taribavirin
(Viramidine).
[0655] Exemplary NS5A inhibitors include, without limitation,
ledipasvir (GS-5885), GS-5816, JNJ-47910382, daclatasvir
(BMS-790052), ABT-267, MK-8742, EDP-239, IDX-719, PPI-668,
GSK-2336805, ACH-3102, A-831, A-689, AZD-2836 (A-831), AZD-7295
(A-689), and BMS-790052.
[0656] Exemplary NS5B inhibitors include, without limitation,
polymerase inhibitor is sofosbuvir (GS-7977), tegobuvir (GS-9190),
GS-9669, TMC647055, ABT-333, ABT-072, setrobuvir (ANA-598),
filibuvir (PF-868554), VX-222, IDX-375, IDX-184, IDX-102,
BI-207127, valopicitabine (NM-283), R1626, PSI-6130 (R1656),
PSI-7851, BCX-4678, nesbuvir (HCV-796), BILB 1941, MK-0608, NM-107,
R7128, VCH-759, GSK625433, XTL-2125, VCH-916, JTK-652, MK-3281,
VBY-708, A848837, GL59728, A-63890, A-48773, A-48547, BC-2329,
BMS-791325, and BILB-1941.
[0657] Exemplary NS3 protease inhibitors include, without
limitation, GS-9451, GS-9256, simeprevir (TMC-435), ABT-450,
boceprevir (SCH-503034), narlaprevir (SCH-900518), vaniprevir
(MK-7009), MK-5172, danoprevir (ITMN-191), sovaprevir (ACH-1625),
neceprevir (ACH-2684), Telaprevir (VX-950), VX-813, VX-500,
faldaprevir (BI-201335), asunaprevir (BMS-650032), BMS-605339,
VBY-376, PHX-1766, YH5531, BILN-2065, and BILN-2061.
[0658] Exemplary alpha-glucosidase 1 inhibitors include, without
limitation, celgosivir (MX-3253), Miglitol, and UT-231B.
[0659] Exemplary hepatoprotectants include, without limitation,
IDN-6556, ME 3738, MitoQ, and LB-84451.
[0660] Exemplary non-nucleoside inhibitors of HCV include, without
limitation, benzimidazole derivatives, benzo-1,2,4-thiadiazine
derivatives, and phenylalanine derivatives.
[0661] Exemplary nucleoside analogues include, without limitation,
ribavirin, viramidine, levovirin, a L-nucleoside, or isatoribine
and said interferon is .alpha.-interferon or pegylated
interferon.
[0662] Exemplary other drugs for treating HCV infection include,
without limitation, imiquimod, 852A, GS-9524, ANA-773, ANA-975,
AZD-8848 (DSP-3025), PF-04878691, and SM-360320, cyclophillin
inhibitors (e.g., DEBIO-025, SCY-635, or NIM811) or HCV IRES
inhibitors (e.g., MCI-067).; emericasan (IDN-6556), ME-3738,
GS-9450 (LB-84451), silibilin, or MitoQ. BAS-100, SPI-452,
PF-4194477, TMC-41629, GS-9350, GS-9585, and roxythromycin.
[0663] Additional exemplary other drugs for treating HCV infection
include, without limitation, zadaxin, nitazoxanide (alinea),
BIVN-401 (virostat), DEBIO-025, VGX-410C, EMZ-702, AVI 4065,
bavituximab, oglufanide, PYN-17, KPE02003002, actilon (CPG-10101),
KRN-7000, civacir, GI-5005, ANA-975 (isatoribine), XTL-6865, ANA
971, NOV-205, tarvacin, EHC-18, and NIM811.
[0664] Still further exemplary other drugs for treating HCV
infection include, without limitation, thymosin alpha 1 (Zadaxin),
nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17 (altirex),
KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000, civacir,
GI-5005, XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971,
NOV-205, tarvacin, EHC-18, VGX-410C, EMZ-702, AVI 4065, BMS-650032,
Bavituximab, MDX-1106 (ONO-4538), Oglufanide, FK-788, VX-497
(merimepodib), DEBIO-025, ANA-975 (isatoribine), XTL-6865, or
NIM811.
General Synthetic Procedures
[0665] The schemes, procedures, and examples provided herein
describe the synthesis of compounds disclosed herein as well as
intermediates used to prepare the compounds. It is to be understood
that individual steps described herein may be combined. It is also
to be understood that separate batches of a compound may be
combined and then carried forth in the next synthetic step.
[0666] The following schemes describe methods that are useful for
preparing compounds disclosed herein.
[0667] L.sub.F is a "linker fragment," (that is to say, a precursor
to L) wherein an attached unsaturated carbon-carbon bond (e.g.
alkene or alkyne) at the portion of L.sub.F distal to {circle
around (T)} facilitates, as a non-limiting example, a metal
catalyzed reaction that results in the connection of L.sub.F to U
to form an L group. Non-limiting examples of metal catalyzed
reactions that result in such a connection include Ru catalyzed
ring closing metathesis or a Pd catalyzed cross coupling reaction
(e.g. Negishi, Heck, or Sonagashira couplings).
[0668] .sup.1H Nuclear magnetic resonance (NMR) spectra were in all
cases consistent with the proposed structures. Characteristic
chemical shifts (6) are given in parts-per-million downfield from
tetramethylsilane using conventional abbreviations for designation
of major peaks: e.g. s, singlet; d, doublet; t, triplet; q,
quartet; m, multiplet; br, broad. The following abbreviations have
been used for common solvents used in nuclear magnetic resonance
experiments: CDCl.sub.3, deuterochloroform; CD.sub.3OD,
perdeuteromethanol; CD.sub.3CN, perdeuteroacetonitrile;
d.sub.6-DMSO, perdeuterodimethylsulfoxide. Mass spectra were
obtained using Thermo Scientific or Agilent Technologies mass
spectrometers equipped with electrospray ionisation (ESI). Masses
are reported as ratios of mass to charge (m/z) of, for example, an
ion of the compound (represented by [M].sup.+), an ion formed from
the compound with another ion, such as a hydrogen ion (represented
by [M+H].sup.+), a sodium ion (represented by [M+Na].sup.+), an ion
formed from the compound by losing an ion, such as the deprotonated
compound (represented by [M-H].sup.-), etc. Analytical HPLC
measurements were performed on Agilent Technologies Series 1100
HPLC using Phenomenex Kinetex C18, 2.6 um 100 .ANG., 4.6.times.100
mm column with an elution program of 2% Solvent B for 0.55 min,
gradient to 98% solvent B over 8 min which is maintained at 98%
solvent B for 0.40 min before returning to 2% solvent B over 0.02
min and maintaining at 2% solvent B for 2.03 min at a flow rate of
1.5 mL/min (Solvent A=MiliQ filtered H.sub.2O+0.1% TFA, Solvent
B=MeCN+0.1% TFA). The term "thin layer chromatography (TLC)" refers
to silica gel chromatography using silica gel 60 F.sub.254 plates.
The retention factor ("R.sub.f") of a compound is the distance
traveled by a compound divided by the distance traveled by the
solvent front on a TLC plate. Terms such as "early eluting" and
"late eluting" refer to the order in which a compound elutes or is
recovered from a solid stationary phase/liquid solvent mobile phase
based chromatography method (e.g. normal phase silica gel
chromatography or reverse phase high pressure liquid chromatography
(HPLC)).
##STR00133##
[0669] Scheme 1 demonstrates a general route to S1-3, where J,
R.sup.1, R, M, L, T, U, W and Q are as defined herein, Z.sup.2a is
as defined in Formula IV or III, or is Z.sup.2 as defined in
Formula I or II. In scheme 1, ester intermediate S1-1 is hydrolyzed
with a base such as lithium hydroxide when R is C.sub.1-C.sub.3
alkyl (e.g., methyl), or with acid such as trifluoroacetic acid
when R is tert-butyl. The product of the ester hydrolysis is then
coupled to an intermediate S1-2 through a coupling reaction (e.g.
using a peptide coupling agent such as HATU and a base such as
DIPEA) to generate compounds of the general structure S1-3.
##STR00134##
[0670] Scheme 2 shows a general synthesis of an intermediate S2-6
where U, W, R.sup.1, J, and Q are as defied herein. In scheme 2, an
appropriately substituted and protected proline species S2-2
undergoes an etherification reaction such as S.sub.NAr (e.g.
treatment with Cs.sub.2CO.sub.3 and S2-1 where R.sup.2 is H and
LG.sup.2 is halogen), S.sub.N2 (e.g. preconversion of S2-2 to a
brosylate (R.sup.2 is Bs) followed by treatment with S2-1 where
LG.sup.2 is --OH and base such as DABCO), Mitsunobu reaction (e.g.
treatment of S2-2 with DIAD and triphenylphosphine followed by S2-1
where LG.sup.2 is --OH) or metal catalyzed cross coupling reaction
(LG.sup.2 is halogen, R.sup.2 is H) to generate intermediate S2-3.
Intermediate S2-3 is deprotected (e.g. 4 N HCl in dioxane when PG
is Boc) to make intermediate S2-4. Amide bond formation via
activation of the carboxylic acid of S2-5 using peptide coupling
agents or other carboxylic acid activation methods prior to
treatment of S2-4 provides intermediate S2-6.
##STR00135## ##STR00136##
[0671] Scheme 3 shows a general synthesis of intermediate S3-6
where L.sub.F-CH.sub.2--CH.sub.2 is L, and U, W, R.sup.1, J, Q, M,
T, and L are as defied herein. In scheme 3, an intermediate S3-1 is
coupled via amide bond formation reaction to an intermediate S3-2
to provide intermediate S3-3. Metal catalyzed cross-coupling (e.g.
Suzuki reaction using potassium vinyltrifluoroborate, Et.sub.3N,
Pd(dppf)Cl.sub.2) to give S3-4, followed by ring closing metathesis
(e.g. Zhan 1B) to give S3-5, followed by reduction of the double
bond (e.g. H.sub.2, 10% Pd/C) provides intermediate S3-6.
##STR00137##
[0672] Scheme 4 shows a general synthesis of an intermediate S4-5
where L.sub.F-CH.sub.2--CH.sub.2 is L, and U, W, R.sup.1, J, Q, Q
and L are as defied herein. In scheme 4, intermediate S4-1 is
protected with a protecting group such as Boc. S4-1 undergoes a
transition metal catalyzed cross coupling (e.g. Sonogashira
coupling) to an intermediate S4-2 to provide intermediate S4-3. The
triple bond of intermediate S4-3 is reduced to a single bond by
hydrogenation (e.g. H.sub.2, catalytic 10% Pd/C) to give
intermediate S4-4. Deprotection of the Boc-amine followed by
coupling under basic conditions (e.g. triethylamine) provides
intermediate S4-5.
##STR00138## ##STR00139##
[0673] Scheme 5 shows a general synthesis of an intermediate S5-9
where L.sub.F-CH.sub.2--CH.sub.2 is L, and U, W, R.sup.1, J, Q, T
and L are as defied herein. In scheme 5 intermediate S5-1 undergoes
a metal catalyzed cross coupling (such as Sonogashira reaction)
with an intermediate S5-2 to provide intermediate S5-3. The triple
bond of intermediate S5-3 is reduced to a single bond under
appropriate conductions such as by hydrogenation (e.g. using
H.sub.2 over catalytic 10% Pd/C) to give intermediate S5-4.
Deprotection of the alcohol to provide S5-5, followed by activation
(e.g. DSC under basic conditions, e.g. triethylamine) provides
intermediate S5-6. Coupling of S5-6 and S5-7 under basic conditions
provides S5-8. Deprotection of the proline nitrogen (e.g. HCl in
dioxane when PG=Boc) followed by a macrolactamization (e.g.
coupling agent such as HATU under basic conditions) provides
intermediate S5-9.
##STR00140##
[0674] Scheme 6 shows a general synthesis of the intermediates S6-6
and S6-7 where U, R.sup.1, J, Q, M, T and L are as defied herein.
In scheme 6 intermediate S6-1, W is OPG, where PG is a protecting
group. S6-1 is first deprotected to give intermediate S6-2.
Alkylation of intermediate S6-2 with an appropriate electrophile
such as S6-4 provides intermediate S6-6. Reaction of S6-2 with
triflic anyhydride provides S6-3, which then undergoes metal
catalyzed cross coupling with an appropriate nucleophilic coupling
partner such as S6-5 (e.g. Sonagashira or Suzuki reaction) to
provide intermediate S6-7.
##STR00141## ##STR00142##
[0675] Scheme 7 shows a general synthesis of intermediate S7-13
where L.sub.F-CH.sub.2--CH.sub.2--CF.sub.2 is L, and W, R.sup.1, J,
Q, M, and T are as defied herein. In S7-13, L is C.sub.1-C.sub.3
alkyl. In Scheme 7, intermediate S7-1 first undergoes lithium
halogen exchange and then is treated with intermediate S7-2 to
generate intermediate S7-3, which is then condensed with
intermediate S7-4 to provide quinoxaline intermediate S7-5.
Halogenation of S7-5 (e.g. POCl.sub.3) provides intermediate S7-6.
Intermediate S7-6 is attached via an ether formation to
intermediate S7-7 through an S.sub.NAr reaction (e.g.
Cs.sub.2CO.sub.3) to generate intermediate S7-8. Deprotection of
the N-PG of intermediate S7-8 provides S7-10. An amide bond
coupling reaction of intermediate S7-9 and intermediate S7-10 (e.g.
EDC and HOBT, or HATU, NMM, DIPEA) provides intermediate S7-11.
Ring closing metathesis of S7-11 generates intermediate S7-12.
Reduction of the double bond (e.g. hydrogenation over palladium on
carbon) provides intermediate S7-13.
##STR00143##
[0676] Scheme 8 shows a general syntheses of intermediate S8-5
wherein an appropriately protected 4-oxo proline S8-1 is reacted
with Bredereck's reagent to generate enaminone S8-2. Addition of an
organometallic species provides enone S8-3, which undergoes
reduction to hydroxyl intermediate S8-4 in a stereoselective manner
(e.g. Luche reduction or CBS reduction). Subsequent olefin
reduction gives 3-substituted hydroxy proline intermediate
S8-5.
##STR00144##
[0677] Scheme 9 shows a general synthesis of intermediate S9-3
wherein a vinyl triflate S9-1 (prepared for example, by methods in
Kamenecka, T. M., et al. Tetrahedron Letters, 2001, 8571) undergoes
metal catalyzed cross coupling (e.g. Negishi coupling) to generate
intermediate S9-2. Hydroboration and subsequent oxidation of
intermediate S9-2 provides intermediate S9-3.
##STR00145##
[0678] Scheme 10 shows a general synthesis of substituted
sulfonamide intermediate S10-3. Tert-butyl
cyclopropylsulfonylcarbamate S10-1 is deprotonated (e.g. n-BuLi)
and reacted with an electrophile (e.g. alkyl halide) to give the
protected substituted sulfonamide intermediate S10-2, which is then
deprotected (e.g. 4 N HCl in dioxane) to provide intermediate
S10-3.
##STR00146##
[0679] Scheme 11 shows a general synthesis of an intermediate S11-3
where E is as defined herein. In Scheme 11, a sulfonamide S11-1 is
coupled to a protected amino acid S11-2 using a coupling agent such
as CDI and a base such as DBU.
##STR00147##
[0680] Scheme 12 shows a general synthesis of intermediates S12-10
and S12-17, where L.sub.F is C.sub.1-C.sub.3 alkylene. In Scheme
12, both syntheses begin with the monoprotection of intermediate
S12-1 to produce S12-2, followed by oxidation (e.g. Swern
oxidation) to provide intermediate S12-3. Enantioselective alpha
chlorination (e.g. organocatalyst S12-4 and NCS) provides
chloroaldehyde S12-5. Reaction of S12-5 with a bis-zinciomethane
derivative (e.g. Nysted's reagent) provides cyclopropane
intermediate S12-6. Intermediate S12-6 is orthogonally protected to
provide intermediate S12-7. Deprotection of --OPG of S12-7 provides
intermediate S12-8, which is subsequently dehydrated (e.g. Grieco's
reagent) to intermediate S12-9 and finally O-PG.sup.2 is removed to
afford intermediate S12-10. Intermediate S12-6 is alternatively be
activated (e.g. DSC and a base such as pyridine) to provide
intermediate S12-11 which is coupled to intermediate S12-12 to
provide carbamate intermediate S12-13. Intermediate S12-13 is
deprotected to give intermediate S12-14, which is then oxidized
(e.g. Swern oxidation) to provide aldehyde intermediate S12-15.
Olefination (e.g. Wittig reaction) of intermediate S12-15 provides
intermediate S12-16. Ester hydrolysis (e.g. LiOH when R is methyl,
TFA when R=tert-butyl) affords intermediate S12-17.
##STR00148##
[0681] Scheme 13 shows a general synthesis of intermediate S13-5
where Q and T are as defined herein and L.sub.F is C.sub.1-C.sub.3
alkylene. Activation of intermediate S13-1 (e.g. DSC) followed by
carbamate formation between intermediate S13-2 and amino acid ester
intermediate S13-3 under basic conditions gives ester intermediate
S13-4. Ester hydrolysis (e.g. LiOH when R=methyl or TFA when
R=tert-butyl) provides intermediate S13-5.
##STR00149##
[0682] Scheme 14 shows a general synthesis of intermediate S14-7
where Q is as defined herein and L.sub.F is C.sub.1-C.sub.3
alkylene. Oxidation of intermediate S14-1 (e.g Dess-Martin
periodinane) produces ketone S14-2. Treatment of S14-2 with S14-3
(e.g. R.sup.2 is --CF.sub.3) in the presence of suitable reagent
(such as CsF) provides intermediate S14-4. Deprotection of S14-4
(e.g. TBAF) provides S14-5, which is then added to an isocyanate
S14-6 to give intermediate S14-7.
##STR00150##
[0683] Scheme 15 shows a general synthesis of an intermediate
(.+-.)-S15-3, generated from the Kulinkovich reaction of a Grignard
reagent S15-1 and an ester S15-2, according to standard procedures
as described in Kulinkovich, O. G. and Kananovich, D. G., Eur. J.
Org. Chem. 2007, 2007, 2121.
##STR00151##
[0684] Scheme 16 shows a general synthesis of an intermediate S16-4
where Q, M, and T are as defined herein and L.sub.F is
C.sub.1-C.sub.3 alkylene. In Scheme 16, olefin S16-1 undergoes
oxidative cleavage (e.g. OsO.sub.4, NalO.sub.4) to aldehyde S16-2,
which is then reduced to alcohol S16-3 (e.g. NaBH.sub.4) and
finally is dehydrated (e.g. Greico elimination) to afford
intermediate S16-4.
##STR00152##
[0685] Scheme 17 shows two general synthetic strategies for
producing intermediate S17-3 where J is as defined herein. In
Scheme 17, an appropriately protected 4-oxo proline S17-1 is
deprotonated and alkylated (e.g. LiHMDS followed by J-LG). A second
deprotonation with base followed by re-protonation at low
temperature generates stereoenriched intermediate S17-2, based on a
described protocol (Blanco, M-J. et. al. J. Org. Chem. 1999, 64,
8786). Reduction of the ketone in a stereoselective manner (e.g.
CBS reduction) provides alcohol S17-3. Where J is methyl, Scheme 17
shows an alternative general synthesis wherein intermediate S17-4
is hydrogenated to generate a mixture of S17-5 and S17-6. Ketone
reduction of S17-5 in a stereoselective manner (e.g. CBS reduction)
provides intermediate S17-3, where J is methyl.
##STR00153##
[0686] Scheme 18 shows a general synthesis of intermediates S18-4
and S18-5, wherein an appropriately protected 4-oxo proline S18-1
is hydroxylated in a stereoselective manner (e.g. MoOPh) to provide
intermediate S18-2, which is subsequently reacted with an
alkylating agent (e.g. trimethyloxonium tetrafluoroborate) to
afford intermediate S18-3. Reduction of the ketone (e.g.
BH.sub.3.SMe.sub.2 complex) provides intermediates S18-4 and
S18-5.
##STR00154##
[0687] Scheme 19 shows a general synthesis of an intermediate S19-7
where Q is as defined herein and L.sub.F is C.sub.1-C.sub.3
alkylene. In Scheme 19, an epoxide intermediate S19-1 is converted
to the (.+-.)-trans-intermediate S19-3. Activation of the alcohol
intermediate (.+-.)-S19-3 (e.g. DSC) produces carbonate
(.+-.)-S19-4, which is treated with intermediate S19-5 to afford
carbamate intermediate S19-6. Intermediate S19-6 then undergoes
ester hydrolysis (e.g. LiOH when R=methyl or TFA when R=tert-butyl)
to provide intermediate S19-7.
##STR00155##
[0688] Scheme 20 shows a general synthesis of an intermediate S20-3
where L.sub.F-O is F, and U, W, R.sup.1, J, Q, M, T and L are as
defied herein. In scheme 20, intermediate S20-1 first undergoes
oxidative cleavage of an olefin (e.g. OsO.sub.4, NaIO.sub.4) and
subsequent reduction of the resultant aldehyde (e.g. NaBH.sub.4) to
provide intermediate S20-2. Transition metal catalyzed cross
coupling provides intermediate S20-3.
##STR00156##
[0689] Scheme 21 shows a general synthesis of an intermediate S21-7
where Q and T are as defined herein. In Scheme 21, activation of
mono-protected diol S21-1 (e.g. DSC) followed by coupling with
amino ester intermediate S21-3 provides carbamate intermediate
S21-4. Intermediate S21-4 is then deprotected to unmask the alcohol
functionality (intermediate S21-5) which is then allylated to
provide intermediate S21-6. Intermediate S21-6 then undergoes ester
hydrolysis (e.g. LiOH when R=methyl or TFA when R=tert-butyl) to
provide intermediate S21-7.
##STR00157##
[0690] Scheme 22 shows a general synthesis of an intermediate S22-3
where U, W, R.sup.1, J, and Q are as defied herein. In scheme 22
intermediate S22-1 is globally deprotected to provide amino acid
intermediate S22-2. The acid functionality of intermediate S22-2 is
then converted to a base-labile carboxylic acid ester (e.g. methyl
ester), intermediate S22-3.
Preparation of Selected Intermediates
Preparation of Intermediate A1.
##STR00158##
[0691] Steps 1-3. Preparation of Intermediate A1: Intermediate A1
was prepared using the procedure detailed in Example 2.12 of
International Patent Publication No. WO 2008/064066 (hereinafter
"WO '066") (p. 75-76) substituting (1R,2S)-methyl
1-(tert-butoxycarbonylamino)-2-vinylcyclopropane-carboxylate
(prepared according to Beaulieu, P. L., et al., J. Org. Chem. 2005,
70, 5869) for (1R,2S)-ethyl
1-(tert-butoxycarbonylamino)-2-vinylcyclopropane-carboxylate.
Preparation of Intermediate A2.
##STR00159##
[0692] Intermediate A2 was prepared similarly to Intermediate A1,
substituting 1-methylcyclopropane-1-sulfonamide (prepared according
to Example 1.2 of WO '066, p. 47) for cyclopropanesulfonamide.
Preparation of Intermediate A3.
##STR00160##
[0693] Step 1. Preparation of A3-1: Cyclopropane ester A3-1 was
prepared from (1R,2S)-methyl
1-(tert-butoxycarbonylamino)-2-vinylcyclopropanecarboxylate
(prepared according to Beaulieu, P. L., et al., J. Org. Chem. 2005,
70, 5869) using the procedure detailed in Example 26 of
International Patent Publication No. WO 2009/005677 (hereinafter
"WO '677") (p. 176). Steps 2-4. Preparation of Intermediate A3:
Intermediate A3 was prepared similarly to
(1R,2S)-1-amino-N-(cyclopropylsulfonyl)-2-vinylcyclopropanecarbox-amide
hydrochloride of Example 2.12 of WO '066 (p. 75-76) substituting
A3-1 for (1R,2S)-ethyl
1-(tert-butoxycarbonylamino)-2-vinylcyclopropane-carboxylate.
Preparation of Intermediate A4.
##STR00161##
[0694] Intermediate A4 was prepared similarly to Intermediate A3,
substituting 1-methylcyclopropane-1-sulfonamide (prepared according
to Example 1.2 of WO '066, p. 47) for cyclopropanesulfonamide.
Preparation of Intermediate A5.
##STR00162##
[0695] Steps 1-3. Preparation of Intermediate A5: Intermediate A5
was prepared similarly to
(1R,2S)-1-amino-N-(cyclopropylsulfonyl)-2-vinylcyclopropane-carboxamide
hydrochloride of Example 2.12 of WO '066 (p. 75-76) substituting
A5-1 (prepared according to Example 104 of WO '677, p. 265) for
(1R,2S)-ethyl
1-(tert-butoxycarbonylamino)-2-vinylcyclopropanecarboxylate.
Preparation of Intermediate A6.
##STR00163##
[0696] Intermediate A6 was prepared similarly to Intermediate A5,
substituting 1-methylcyclopropane-1-sulfonamide (prepared according
to Example 1.2 of WO '066, p. 47) for cyclopropanesulfonamide.
Preparation of Intermediate A7.
##STR00164##
[0698] Intermediate A7 was prepared according to Example 97.1.6 of
U.S. Patent Publication No. 2009/274652 (hereinafter "US '652"), p.
72-73.
Preparation of Intermediate A8.
##STR00165##
[0700] Steps 1-2. Preparation of Intermediate A8: Intermediate A8
was prepared similarly to
(1R,2S)-1-amino-N-(cyclopropylsulfonyl)-2-vinylcyclopropane-carboxamide
hydrochloride of Example 2.12 of WO '066 (p. 75-76) substituting
A8-1 (prepared according to the procedure detailed in Example
97.1.4 of US '652, p. 72-3) for
(1R,2S)-1-(tert-butoxycarbonylamino)-2-vinylcyclo-propanecarboxylic
acid and substituting 1-methylcyclopropane-1-sulfonamide (prepared
according to Example 1.2 of WO '066, p. 47) for
cyclopropanesulfonamide. A8-1 .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 9.22 (br s, 1H), 6.05-5.75 (m, 1H), 5.38 (br s, 1H), 2.04
(m, 2H), 1.68 (m, 2H), 1.61 (m, 3H), 1.52 (m, 9H), 1.42 (m, 1H),
1.28 (m, 1H), 0.85 (m, 2H).
Preparation of Intermediate A9.
##STR00166##
[0701] Step 1-2. Preparation of Intermediate A9: Intermediate A9
was prepared similarly to
(1R,2S)-1-amino-N-(cyclopropylsulfonyl)-2-vinylcyclopropane-carboxamide
hydrochloride of Example 2.12 of WO '066 (p. 75-76) substituting
A9-1 (prepared according to Example 1, Steps 1L-1O of International
Patent Publication No. WO 2009/134987, p. 75-77) for
(1R,2S)-1-(tert-butoxycarbonylamino)-2-vinylcyclopropanecarboxylic
acid.
Preparation of Intermediate A10.
##STR00167##
[0702] Intermediate A10 was prepared similarly to Intermediate A9,
substituting 1-methylcyclopropane-1-sulfonamide (prepared according
to Example 1.2 of WO '066, p. 47) for cyclopropanesulfonamide.
Preparation of Intermediate A11.
##STR00168## ##STR00169##
[0703] Step 1. Preparation of A11-1: To a solution of NaOH (46.2 g,
50% w/w in water) at rt was added BnEt.sub.3NCl (10.5 g, 46 mmol),
di-tert-butyl malonate (10 g, 46 mmol) and 1,2-dibromopropane (14
g, 69.3 mmol). The mixture was stirred at rt overnight and was
extracted with DCM (3.times.100 mL). The organic layers were washed
with water (80 mL) and brine (50 mL), dried over anhydrous
Na.sub.2SO.sub.4. Concentration in vacuo produced A11-1 that was
used subsequently without further purification. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 1.83-1.62 (m, 1H); 1.42 (s, 9H); 1.40 (s,
9H); 1.24-1.05 (m, 2H); 1.03-1.02 (d, 3H). Step 2. Preparation of
A11-2: To a mixture of t-BuOK (175 g, 1.56 mol) in ether (1.2 L) at
0.degree. C. was added water (3.4 mL) followed by addition of
diester A11-1 (91 g, 0.35 mol). The mixture was stirred at rt for
three days, then quenched with ice-water. The aqueous layer was
extracted with ether (2.times.400 mL), acidified with critic acid,
and then extracted with EA (3.times.400 mL). The combined ethyl
acetate extracts were washed with water (2.times.100 mL), brine
(200 mL), dried over anhydrous Na.sub.2SO.sub.4, and concentrated
in vacuo to produce A11-2 that was used subsequently without
further purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
12.60 (s, 1H); 1.70-1.64 (s, 1H); 1.37 (s, 9H); 1.19-1.13 (m, 1H);
1.03-1.00 (m, 4H). Step 3. Preparation of A11-3: To a mixture A11-2
(33.5 g, 0.17 mol) and triethylamine (70 mL) in THF (200 mL) at
0.degree. C. was added ethyl chloroformate (22 mL). The mixture was
stirred at 0.degree. C. for 1 h. To the mixture at 0.degree. C. was
added sodium azide (54 g, 0.83 mol, 4.9 eq) in water (100 mL), the
mixture was stirred for 40 min. The mixture was extracted with EA
(2.times.400 mL), washed with water (100 mL), brine (100 mL), dried
over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo to
produce a residue that was taken up in toluene (100 mL) and treated
with benzyl alcohol (50 mL). The mixture was then heated at
70.degree. C. for 2 h, cooled to rt, adjusted to pH 8 with sodium
bicarbonate, and then extracted with ether (3.times.200 mL). The
aqueous layer was then adjusted to pH 5 with 1 N HCl and extracted
with EA (2.times.300 mL). The combined ethyl acetate extracts were
washed with water (100 mL), brine (80 mL), dried over anhydrous
Na.sub.2SO.sub.4, and concentrated in vacuo to give CBZ protected
amine A11-3 (16 g) that is used subsequently without further
purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.85 (s,
1H); 7.28-7.15 (m, 5H); 4.97-5.03 (m, 2H); 1.33 (s, 9H); 1.33-1.17
(m, 2H); 1.10 (d, J=6.8 Hz, 3H); 0.90-1.00 (m, 1H). Steps 4 and 5.
Preparation of A11-4: To a solution of Cbz protected amine A11-3
(16 g, 52 mmol) in DCM (250 mL) was added dropwise TFA (250 mL,
3.24 mol) at rt and the mixture stirred at rt overnight. The
mixture was concentrated in vacuo, adjusted to pH 8-9 using aqueous
sodium carbonate and washed with ether (3.times.80 mL). The aqueous
phase was then adjusted to pH 5-6 using 1 N HCl and extracted with
EA (2.times.300 mL). The combined ethyl acetate phases were washed
with water (80 mL), brine (80 mL), dried over anhydrous
Na.sub.2SO.sub.4 and concentrated to give 13 g as a slightly yellow
oil that was used in the next step without further purification.
This material (8.0 g, 32 mmol) was taken up in methanol (200 mL),
treated with thionyl chloride (15 mL) at 0.degree. C., then stirred
at rt overnight. The resulting mixture was concentrated in vacuo
and purified by flash chromatography on silica (eluent PE/EA
10:1-5:1) to give methyl ester A11-4 (6 g). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.97 (s, 1H); 7.37-7.26 (m, 5H); 4.99 (s,
2H); 3.61 (s, 3H); 1.48-1.45 (m, 1H); 1.17-1.08 (m, 2H); 1.06-1.04
(d, 3H). Step 6. Preparation of A11-5: Cbz carboxamide A11-4 (36 g,
0.15 mol), Boc.sub.2O (40 g, 0.18 mol), and Pd/C (3.6 g, 10% w/w)
were combined in methanol under H.sub.2 and stirred at 32.degree.
C. overnight. The reaction mixture was filtered to remove the
catalyst, additional Boc.sub.2O (40 g, 0.18 mol) and Pd/C (3.6 g,
10% w/w) were added and the reaction placed under a H.sub.2
atmosphere with stirring at rt for a weekend. The reaction mixture
was filtered to remove the catalyst, concentrated in vacuo and
purified by flash chromatography on silica (eluent PE/EA 20:1-10:1)
to produce Boc protected amine A11-5. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 7.48 (s, 1H), 3.59 (s, 3H), 1.43-1.41 (m,
1H), 1.34 (s, 9H), 1.21-1.18 (m, 1H), 1.07-1.01 (m, 4H). Step 7.
Preparation of A11-6: To a solution of NaH.sub.2PO.sub.4 (1.9 g) in
water (160 mL) at 40.degree. C. was added Alcalase (2.4 U/g, 16
mL). The mixture was adjusted with 50% aqueous sodium hydroxide to
pH 8. A11-5 (2.80 g) in DMSO (32 mL) was added to the buffer
dropwise over 30 min. The mixture was stirred at 40.degree. C. and
maintained at pH 8 with addition of 50% NaOH for 19 h. The mixture
was cooled to rt, with ether (3.times.100 mL) and the organic phase
washed with sat. NaHCO.sub.3 (2.times.40 mL), water (2.times.40
mL), brine (40 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo to produce A11-6. .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 5.18 (br s, 1H); 3.71 (s, 3H); 1.43-1.18 (m,
2H); 1.34 (s, 9H); 1.07-1.01 (m, 4H). Analysis of the product using
chromegaChiral CC3 column (0.46 cm I.D..times.25 cm L, 3 .mu.L
injection, 80/20 hexane/IPA, 1 mL/min, 34.degree. C., 220 nM UV
detection) determined the enantiomeric excess was 99.4% (desired
RT=5.238 min, undesired RT=6.745 min). Steps 8 and 9. Preparation
of A11-7: Solid LiOH*H.sub.2O (19.1 g, 455 mmol) is taken up in 50
mL MeOH/50 mL water at rt. Once all LiOH has dissolved, methyl
ester A11-6 (10.4 g, 45.5 mmol) is taken up in 100 mL THF added to
reaction mixture and stirred vigorously overnight. The resulting
solution is diluted with water (150 mL), adjusted to pH 3 with 12 M
HCl and extracted with EtOAc. The combined organic layers are
washed with brine, dried over anhydrous MgSO.sub.4 and concentrated
in vacuo to produce a fine white powder (9.2 g). This material (1.5
g, 7 mmol) is taken up in THF (30 mL) and treated with CDI (1.47 g,
9.1 mmol). The resulting solution was heated to 65.degree. C. for 2
h, cooled to rt and treated with DBU (2.1 mL, 13.9 mmol) and
1-methylcyclopropane-1-sulfonamide (1.4 g, 10.5 mmol). The
resulting solution is stirred at rt overnight. Addition of 1 M HCl
is used to adjust the pH .about.1 prior to removing the majority of
THF in vacuo. The resulting slurry is extracted with EtOAc and the
combined organics washed with brine, dried over anhydrous
MgSO.sub.4 and concentrated in vacuo to produce 2.29 g of acyl
sulfonamide A11-7. LCMS-ESI (m/z): [M+Na].sup.+ calcd for
C.sub.14H.sub.24N.sub.2NaO.sub.5S: 355.41. found: 355.84. Step 10.
Preparation of Intermediate A11. Acyl sulfonamide A11-7 (0.25 g,
0.75 mmol) in dioxane (1 mL) is treated with HCl (4 M in dioxane,
2.8 mL, 11.2 mmol) at rt. After 4 h, the reaction is concentrated
in vacuo to produce 0.20 g of Intermediate A-11 that is used
subsequently without additional purification. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 1.87-1.84 (m, 0.5; H); 1.77-1.65 (m, 1.5H);
1.58-1.46 (m, 2H); 1.54 (d, J=8 Hz, 3H); 1.34-1.26 (m, 3+1H);
1.02-0.92 (m, 1H); 0.83-0.77 (m, 1H).
Preparation of Intermediate A12.
##STR00170##
[0704] Step 1. Preparation of A12-1: A vessel containing a solution
of carboxylic acid A9-1 (1 g, 4 mmol) in THF (15 mL) was treated
with CDI (0.84 g, 5.2 mmol), sealed and heated to 75.degree. C. for
2 h. The clear tan colored solution is divided in half and used
subsequently without further purification for the remainder of Step
1 in the preparation of Intermediate A12 as well as the preparation
of Intermediate A13 as detailed below. This solution is treated
with 1-fluorocyclopropane-1-sulfonamide (0.42 g, 3 mmol; prepared
according to Steps 1, 4, and 9 of Example 7 of International Patent
Publication No. WO 2009/14730, p. 107-110) and DBU (0.6 mL, 4 mmol)
and allowed to stir overnight at rt. The solution was acidified to
pH .about.1 with 1 M HCl and concentrated in vacuo to remove the
majority of THF. The aqueous layer was extracted with EtOAc and the
combined organics washed with brine, dried over anhydrous
MgSO.sub.4 and concentrated in vacuo to dryness to afford 0.73 g of
the A12-1 that was used without further purification. Step 2.
Preparation of Intermediate A12: Acyl sulfonamide A12-1 (0.25 g,
0.67 mmol) was taken up in 1 mL dioxane and treated with HCl (4 M
in dioxane, 2.5 mL, 11 mmol). The reaction was stirred at rt for 2
h and concentrated in vacuo to dryness to afford a quantitative
yield of Intermediate A12. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 6.04 (td, J.sub.H-F=55.6 Hz, J=5.2 Hz, 1H); 2.25-2.14 (m,
1H); 1.78-1.62 (m, 2H); 1.52-1.38 (m, 4H).
Preparation of Intermediate A13.
##STR00171##
[0705] Intermediate A13 was prepared similarly to Intermediate A12,
substituting 1-chlorocyclopropane-1-sulfonamide (prepared according
to Li, J, et al. Synlett, 2006, 5, pp. 725-728) for
1-chlorocyclopropane-1-sulfonamide in Step 1. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 6.03 (td, J.sub.H-F=54.8 Hz, J=6 Hz, 1H);
2.32-2.18 (m, 1H); 2.06-1.92 (m, 2H); 1.80-1.68 (m, 2+1H);
1.56-1.44 (m, 1H); 1.44-1.37 (m, 1H).
Preparation of Intermediate B1.
##STR00172##
[0706] Steps 1 and 2. Preparation of Intermediate B1: Enaminone
B1-1 (4.0 g, 11.8 mmol, prepared according to Camplo, M., et al.
Tetrahedron 2005, 61, 3725) was dissolved in acetone (120 mL) and
the reaction vessel was purged with Ar. Pd/C (10 wt. % Pd, 820 mg)
was added in a single portion and the reaction vessel was purged
twice with H.sub.2. The reaction was stirred under 1 atm H.sub.2 at
rt for 15 h and was then filtered through a pad of Celite with
acetone. The filtrate was concentrated and filtered through a plug
of silica gel with 30% EtOAc in hexanes to afford a .about.2:1
mixture of ketones B1-2 and B1-3 (3.48 g) as a white solid. This
mixture (3.37 g, 11.3 mmol) was dissolved in THF (100 mL) under Ar.
A 1 M solution of (R)-(+)-2-methyl-CBS-oxazaborolidine in toluene
(11.3 mL, 11.3 mmol) was added in a single portion and the
resulting solution was cooled to -78.degree. C. A 1 M solution of
BH.sub.3.SMe.sub.2 in CH.sub.2Cl.sub.2 (11.3 mL) was then added
dropwise over 5 min. The resulting solution was stirred for 20 min
and was removed from the cold bath. After an additional 15 min, the
reaction was placed in a water bath at ambient temperature. After
an additional 7 min, the reaction was quenched by dropwise addition
of MeOH (20 mL). After stirring an additional 2.5 h, the reaction
mixture was concentrated, dissolved in EtOAc (300 mL), and washed
with 0.2 M HCl (200 mL). The phases were separated, and the aqueous
phase was extracted with EtOAc (100 mL). The combined organic phase
was filtered to remove solids, dried over Na.sub.2SO.sub.4,
filtered, and concentrated. The crude residue was dissolved in
CH.sub.2Cl.sub.2 and was concentrated onto 20 g silica gel.
Purification by silica gel chromatography (25 to 40% EtOAc in
hexanes) provided partial separation of Intermediate B1 from other
diastereomeric products. Mixed fractions were pooled and
concentrated onto 9 g silica gel. Purification by silica gel
chromatography provided Intermediate B1 contaminated with minor
diastereomeric components as a white solid (1.96 g). .sup.1H NMR
(400 MHz, CDCl.sub.3, rotamers observed) .delta. 4.25-4.15 (m, 1H),
4.13-4.04 (m, 1H), 3.91-3.79 (m, 1H), 3.28-3.09 (m, 1H), 2.41-2.23
(m, 1H), 2.04 (bs, 1H), 1.51-1.39 (m, 18H), 1.09-1.01 (m, 3H).
Preparation of Intermediate B2.
##STR00173##
[0707] Steps 1 and 2. Preparation of B2-1:
trans-3-Hydroxy-L-proline (571 mg, 4.35 mmol, Chem-Impex
International, Inc.) was suspended in MeOH and cooled to 0.degree.
C. Thionyl chloride (1.6 mL, 22 mmol) was added over 5 min and the
solution was warmed to rt. After stirring for 24 h, the reaction
mixture was concentrated under reduced pressure to afford the
methyl ester, which was carried on without further purification.
The crude ester was suspended in DCM (22 mL) and treated with TEA
(1.3 mL, 9.57 mmol). The stirred mixture was cooled to 0.degree. C.
and trityl chloride (1.21 g, 4.35 mmol) was added. The reaction
mixture was allowed to gradually come to rt o/n, and then poured
into saturated aqueous NaHCO.sub.3. The aqueous layer was extracted
three times with DCM. The combined organics were dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The crude residue was purified by silica gel chromatography (25% to
50% EtOAc/Hex to afford alcohol B2-1 (1.27 g). Step 3. Preparation
of B2-2: Alcohol B2-1 (1.23 g, 3.18 mmol) and 2 g 4 .ANG. MS were
suspended in DCM (16 mL) and treated with NMO (560 mg, 4.78 mmol)
and TPAP (76 mg, 0.218 mmol). After stirring for 30 min, the
mixture was filtered over a short pad of silica and eluted off with
50% EtOAc/Hex. The filtrate was concentrated and the crude residue
was purified by silica gel chromatography (10% to 30% EtOAc/Hex to
afford ketone B2-2 (0.99 g). Step 4. Preparation of B2-3: LiHMDS
(1.0 M in THF, 5.8 mL, 5.8 mmol) was added to THF (22 mL) and the
stirred solution was cooled to -78.degree. C. A rt solution of
ketone B2-2 (2.14 g, 5.55 mmol) in THF (6 mL) was added dropwise by
cannula over 5 min. The flask that had contained B2-2 was then
rinsed with THF (4 mL) and the rinsing was added dropwise by
cannula to the reaction mixture. After 35 min,
N-(5-chloro-2-pyridyl)bis(trifluoromethanesulfonimide) (2.40 g,
6.11 mmol) in THF (6 mL) was added to the reaction mixture dropwise
by syringe over 5 min. After another 1 h, the reaction mixture was
warmed to rt. Following an additional 30 min, the reaction was
quenched by addition of 20 mL H.sub.2O and diluted with Et.sub.2O.
The organic solution was washed with 10% NaOH and dried over
K.sub.2CO.sub.3, filtered and concentrated under reduced pressure.
The crude residue was loaded onto a silica column that had been
pre-equilibrated with 1% TEA/Hex. The material was purified by
silica gel chromatography (0% to 15% EtOAc/Hex doped with 1% TEA)
to afford enol triflate B2-3 (1.89 g). Step 5. Preparation of B2-4:
Enol triflate B2-3 (957 mg, 1.85 mmol) was dissolved in THF (9 mL)
and treated with Pd(PPh.sub.3).sub.4(107 mg, 0.0925 mmol) and
dimethyl zinc (2.0 M in PhMe, 1.9 mL, 3.7 mmol). The reaction
mixture was stirred at rt for 5 h, then more dimethyl zinc (2.0 M
in PhMe, 1.9 mL, 3.7 mmol) was added and the reaction was heated to
50.degree. C. for 15 min. After cooling to rt, the mixture was
diluted with Et.sub.2O. The organic solution was washed with 10%
NaOH twice, then dried over MgSO.sub.4, filtered and concentrated
under reduced pressure. The crude B2-4 residue was carried on
without further purification. Steps 6 and 7. Preparation of B2-5:
Compound B2-4 (1.85 mmol theoretical) was dissolved in 1:1 MeOH/DCM
(20 mL) and treated with HCl (4.0 M in dioxane, 2 mL, 8.0 mmol).
After stirring for 2 h at rt, the reaction mixture was concentrated
and the crude material was carried on without further purification.
The crude product amine hydrochloride was treated with Boc.sub.2O
(2.02 g, 9.25 mmol), DCM (18 mL), MeOH (1.8 mL) and TEA (0.52 mL,
3.7 mmol). After stirring for 2 h at rt, the reaction mixture was
diluted with EtOAc and washed with 10% HCl, saturated aqueous
NaHCO.sub.3 and brine. The organic solution was dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
residue was purified by silica gel chromatography (15% to 40%
EtOAc/Hex) to afford carbamate B2-5 (331 mg). LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.12H.sub.20NO.sub.4: 242.14. found:
243.26. Step 8. Preparation of Intermediate B2: Carbamate B2-5 (345
mg, 1.43 mmol) was dissolved in THF (7 mL) and cooled to 0.degree.
C. BH.sub.3.SMe.sub.2 complex (2.0 M in THF, 0.79 mL, 1.58 mmol)
was added dropwise and the reaction mixture was allowed to come to
rt gradually. After 15 h, the reaction was quenched by dropwise
addition of H.sub.2O (added until bubbling ceased), then cooled to
0.degree. C. Hydrogen peroxide (30% w/w in H.sub.2O, 0.73 mL, 7.2
mmol) and NaOH (2.0 M in H.sub.2O, 0.86 mL, 1.72 mmol) were added
in quick succession and the stirred mixture was heated to
50.degree. C. for 35 min. The mixture was then diluted with
Et.sub.2O and washed successively with H.sub.2O, saturated aqueous
NaHCO.sub.3 and brine, then dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. Intermediate B2 was used in
subsequent reactions without further purification. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for C.sub.12H.sub.22NO.sub.5: 260.15.
found: 259.99.
Preparation of Intermediate B3.
##STR00174##
[0708] Step 1. Preparation of B3-1: Enol triflate B2-3 (91 mg,
0.176 mmol) was dissolved in THF (1.7 mL) and treated with
cyclopropyl zinc bromide (0.5 M in THF, 1.7 mL, 0.85 mmol) and
Pd(PPh.sub.3).sub.4 (20 mg, 0.018 mmol). The stirred reaction
mixture was heated to 50.degree. C. for 2 h then cooled to rt and
diluted with EtOAc. The organic solution was washed successively
with saturated aqueous NaHCO.sub.3 and brine, then dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
crude residue was purified by silica gel chromatography (0% to 20%
EtOAc/Hex) to afford cyclopropane B3-1 (43 mg). LCMS-ESI.sup.+
(m/z): [M-Tr+H].sup.+ calcd for C.sub.9H.sub.14NO.sub.2: 168.10.
found: 168.04. Steps 2 and 3. Preparation of B3-2: Vinyl
cyclopropane B3-1 (43 mg, 0.11 mmol) was dissolved in 1:1 MeOH/DCM
(10 mL) and treated with HCl (4.0 M in dioxane, 1 mL, 4.0 mmol).
After stirring for 1.5 h at rt, the reaction mixture was
concentrated and the crude material was carried on without further
purification. The crude product of step 2 was treated with
Boc.sub.2O (229 mg, 1.05 mmol), DMAP (13 mg, 0.105 mmol), DCM (5
mL) and TEA (0.293 mL, 2.10 mmol). After stirring for 5 h at rt,
the reaction mixture was diluted with EtOAc and washed with 10%
HCl, saturated aqueous NaHCO.sub.3 twice and brine. The organic
solution was dried over MgSO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by silica gel
chromatography (10% to 30% EtOAc/Hex) to afford carbamate B3-2 (20
mg). LCMS-ESI.sup.+ (m/z): [M-(t-Bu)+H].sup.+ calcd for
C.sub.10H.sub.14NO.sub.4: 212.09. found: 211.91. Step 4.
Preparation of Intermediate B3: Carbamate B3-2 (152 mg, 0.569 mmol)
was dissolved in THF (5.7 mL) and cooled to 0.degree. C.
BH.sub.3*SMe.sub.2 complex (2.0 M in THF, 0.31 mL, 0.63 mmol) was
added dropwise and the reaction mixture was allowed to come to rt
gradually. After 20 h, the reaction was quenched by dropwise
addition of H.sub.2O (added until bubbling ceased), then cooled to
0.degree. C. Hydrogen peroxide (30% w/w in H.sub.2O, 0.29 mL, 2.85
mmol) and NaOH (2.0 M in H.sub.2O, 0.43 mL, 0.86 mmol) were added
in quick succession and the stirred mixture was heated to
50.degree. C. for 30 min. The mixture was then diluted with
Et.sub.2O and washed successively with H.sub.2O, saturated aqueous
NaHCO.sub.3 and brine, then dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. Intermediate B3 was carried on
without further purification. LCMS-ESI.sup.+ (m/z):
[M-(t-Bu)+H].sup.+ calcd for C.sub.10H.sub.16NO.sub.5: 230.10.
found: 230.03.
Preparation of Intermediate B4.
##STR00175##
[0710] Intermediate B4 ((2S,3S,4R)-di-tert-butyl
3-ethyl-4-hydroxypyrrolidine-1,2-dicarboxylate) was prepared
according to Camplo, M., et al. Tetrahedron 2005, 61, 3725.
Preparation of Intermediate B5.
##STR00176##
[0711] Step 1. Preparation of enone B5-2: To a solution of B1-1 in
tetrahydrofuran (7.35 mL) was added ethylmagnesium bromide (3 M in
diethyl ether, 1.47 mL 4.41 mmol) via syringe at -78.degree. C.
under an argon atmosphere. After 2.5 h, the reaction mixture was
allowed to warm to rt over 30 min at which point the reaction
mixture was diluted with saturated aqueous ammonium chloride
solution (20 mL). The resulting mixture was extracted with ethyl
acetate (20 mL twice), and the combined organic extracts were dried
over anhydrous sodium sulfate and were concentrated in vacuo. The
crude residue was purified by silica gel chromatography (0-100%
ethyl acetate/hexanes gradient) to afford intermediate B5-1 (308.8
mg) as a colorless oil. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.17H.sub.28NO.sub.5: 326.2. found: 326.2. Step 2. Preparation
of B5-2: To a solution of enone B5-1 (308 mg, 0.95 mmol) in
methanol (4.7 mL) was added cerium(III) chloride heptahydrate (566
mg, 1.52 mmol) at rt under an argon atmosphere. The resulting
mixture was cooled to -78.degree. C., and sodium borohydride (57.7
mg, 1.52 mmol) was added as a solid. After 1 h, the reaction
mixture was warmed to 0.degree. C. and saturated aqueous ammonium
chloride (20 mL) was added. The resulting mixture was extracted
with ethyl acetate (20 mL twice), and the combined organic extracts
were dried over anhydrous sodium sulfate and were concentrated in
vacuo to afford allylic alcohol B5-2 (319.3 mg) as a colorless oil,
which was used directly in the next step without purification.
LCMS-ESI (m/z): [M+H].sup.+ calcd for C.sub.17H.sub.29NO.sub.5:
328.2. found: 328.2. Step 3. Preparation of Intermediate B5: To a
solution of alcohol B5-2 (319 mg, 0.98 mmol) in ethanol (4.9 mL)
was added Pd/C (10%, 103.9 mg, 0.097 mmol) at rt under an argon
atmosphere. The atmosphere was replaced with hydrogen and the
reaction mixture was stirred vigorously at rt. After 16 h, the
reaction mixture was diluted with ethyl acetate (25 mL) and was
filtered through a pad of Celite with ethyl acetate washings (10 mL
three times). The filtrate was concentrated in vacuo to afford
Intermediate B5 (188 mg), which was used directly in the next step
without purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.17H.sub.32NO.sub.5: 330.2. found: 330.3.
Preparation of Intermediate B6.
##STR00177##
[0712] Step 1. Preparation of B6-1: A solution of
isopropylmagnesium bromide (2.9 M in MeTHF, 3.2 mL, 9.3 mmol) was
added dropwise to a cooled solution of B1-1 (1.02 g, 3.00 mmol) in
60 mL of ether at -78.degree. C. under argon. Reaction mixture was
warmed to room temperature and stirred for 3 hours. Reaction
mixture was quenched with sat. aqueous NH.sub.4Cl and extracted
three times with ether. Combined organics were washed with sat.
aqueous NaHCO.sub.3 and brine, dried (MgSO.sub.4), filtered, and
concentrated under reduced pressure. The resulting residue was
purified by silica gel chromatography (0-30% ethyl acetate in
hexanes) to yield B6-1 (743 mg) as a light yellow oil. .sup.1H NMR
(400 MHz, CDCl.sub.3): .delta. 6.60 (dd, J=10.8, 2.4 Hz, 1H), 5.14
and 5.06 (rotamers, d, J=2.4 Hz, 1H), 3.96 (m, 2H), 2.91 (m, 1H),
1.46 (s, 9H), 1.27 (s, 9H), 1.04 (d, J=8.8 Hz, 6H). Step 2.
Preparation of B6-2 and B6-3: CeCl.sub.3.7H.sub.2O (1.32 g, 3.50
mmol) was added to a solution of B6-1 (740 mg, 2.18 mmol) in 47 mL
of methanol at room temperature under argon. After cooling to
-78.degree. C., sodium borohydride (127 mg, 3.34 mmol) was added
slowly portionwise. After two hours, reaction mixture was warmed to
0.degree. C. After fifteen minutes, reaction mixture was quenched
with sat. aqueous NH.sub.4Cl and extracted three times with ethyl
acetate. Combined organics were washed with brine, dried
(MgSO.sub.4), filtered, and concentrated under reduced pressure to
yield a .about.3:1 mixture of B6-2 (major) and B6-3 (minor) as a
colorless film (738 mg), which was used in the next step without
further purification. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
5.68-5.48 (m, 1H), 4.90-4.31 (m, 2H), 4.05-3.15 (m, 2H), 2.90-2.61
(m, 1H), 1.50-1.39 (br s, 18H), 1.02 (d, J=9.2 Hz, 6H). Step 3.
Preparation of Intermediate B6: The .about.3:1 mixture of B6-2 and
B6-3 (341 mg, 1.00 mmol) was dissolved in 28 mL of ethyl acetate.
Palladium on carbon (10 wt %, 109 mg, 0.11 mmol) was then added and
mixture was hydrogenated under an atmosphere of hydrogen for
nineteen hours. Mixture was then filtered over Celite, washing with
ethyl acetate, and filtrate was concentrated under reduced
pressure. The resulting residue was purified by silica gel
chromatography (0-50% ethyl acetate in hexanes) to yield
Intermediate B6 (141 mg) as a colorless oil. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 4.31-4.17 (m, 2H), 3.97-3.85 (m, 1H),
3.21-3.07 (m, 1H), 2.35-2.18 (m, 1H), 1.92-1.78 (m, 1H), 1.47-1.37
(m, 18H), 1.35-1.19 (m, 2H), 0.94 (d, J=8.8 Hz, 6H).
Preparation of Intermediate B7.
##STR00178##
[0713] Step 1. Preparation of B7-2: To a solution of alcohol B7-1
(500 mg, 1.33 mmol; prepared according to Barreling, P., et al.
Tetrahedron 1995, 51, 4195) in DCM (6.65 mL) was added Dess-Martin
periodinane (564 mg, 1.33 mmol) at rt under an argon atmosphere.
After 2 h, the reaction mixture was purified directly by silica gel
chromatography (0-100% ethyl acetate/hexanes gradient) to afford
ketone B7-2 (431 mg) as a colorless oil. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.21H.sub.30NO.sub.5: 376.2. found:
376.2. Step 2. Preparation of Intermediate B7: To a solution of
intermediate B7-2 (410 mg, 1.09 mmol) and
(R)-(+)-2-methyl-CBS-oxazaborolidine (Aldrich, 1M in toluene, 1.09
mL, 1.09 mmol) in THF (5.45 mL) was added BH.sub.3.THF (1M in
toluene, 2.18 mL, 2.18 mmol) at -78.degree. C. under an argon
atmosphere. After 1 h, the reaction mixture was quenched with
saturated aqueous ammonium chloride solution (15 mL) and the
resulting mixture was allowed to warm to rt. The phases where
separated and the aqueous phase was extracted twice (20 mL) with
DCM. The combined organic layers were dried over anhydrous sodium
sulfate, and were concentrated in vacuo. The crude residue was
purified by silica gel chromatography (0-100% ethyl acetate/hexanes
gradient) to afford Intermediate B7 (390.9 mg, 4:1 diastereomeric
mixture) as a colorless oil. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.21H.sub.32NO.sub.5: 378.2. found: 378.5.
Preparation of Intermediate B8.
##STR00179##
[0714] Step 1. Preparation of B8-1. n-BuLi (0.44 mL, 1.1 mmol, 2.5M
in hexane) was added to a cold (-78.degree. C.) solution of
(S)-methyl
4-oxo-1-(9-phenyl-9H-fluoren-9-yl)pyrrolidine-2-carboxylate (383
mg, 1 mmol, prepared as described in Sardina, F. J., Blanco, M.-J.
J. Org. Chem. 1996, 61, 4748) in THF/HMPA (3.8 mL/0.4 mL). The
resulting solution was stirred at -78.degree. C. to -50.degree. C.
for 1.5 h, and then bromoacetonitrile (0.2 mL, 3 mmol) was added.
The reaction mixture was stirred while the temperature was allowed
to reach -10.degree. C. (4 h). To the reaction mixture was charged
with saturated aqueous NH.sub.4Cl (1 mL) and EtOAc (15 mL). The
organic layer was separated, and the aqueous layer was extracted
with EtOAc (10 mL). Both organic layers were combined, washed with
H.sub.2O and brine, and dried over Na.sub.2SO.sub.4. The organic
layer was concentrated and purified via silica gel chromatography
to afford diastereomeric mixture B8-1 (170 mg) as colorless oil.
Step 2. Preparation of B8-2. KHMDS (0.4 mL, 0.4 mmol, 1M in THF)
was added to a cold (-78.degree. C.) solution of B8-1 (140 mg, 0.33
mmol) in THF/DMPU (1.5 mL/0.75 mL). The resulting solution was
stirred at -78.degree. C. for 1.5 h. Then HOAc (0.1 mL) was added.
To the reaction mixture was charged with saturated aqueous
NH.sub.4Cl (1 mL) and EtOAc (15 mL). The organic layer was
separated, and the aqueous layer was extracted with EtOAc (10 mL).
Both organic layers were combined, washed with H.sub.2O and brine,
and dried over Na.sub.2SO.sub.4. The organic layer was concentrated
and purified via silica gel chromatography to afford ketone B8-2
(120 mg) as colorless oil. Step 3. Preparation of Intermediate B8.
To an oven-dried, nitrogen-flushed flask was added BH.sub.3.THF
(0.28 mL, 0.28 mmol,) followed by
(R)-(+)-2-methyl-CBS-oxazaborolidine (0.012 mL, 0.03 mmol, 1.0 M in
toluene). A solution of B8-2 (120 mg, 0.28 mmol) in THF (0.5 mL)
was added dropwise. The reaction mixture was stirred at rt for 60
min, and then quenched by addition of 1.0 M aqueous HCl (0.2 mL).
EtOAc (20 mL) was added and organic phase washed with sat. aqueous
NaHCO.sub.3 and brine, and dried over Na.sub.2SO.sub.4. The organic
layer was concentrated and purified via silica gel chromatography
to afford Intermediate B8 (100 mg) as colorless oil. LCMS-ESI
(m/z): [M].sup.+ calcd for C.sub.27H.sub.24N.sub.2O.sub.3: 424.49.
found: 424.77.
Preparation of Intermediate C1.
##STR00180##
[0715] Methyl 3-methyl-N-(oxomethylene)-L-valinate (Intermediate
C1) was prepared according to Step 3 of Intermediate B1 of
International Patent Publication No. WO 2010/11566 (hereinafter
"WO'566"), p 14.
Preparation of Intermediate C2.
##STR00181##
[0716] Intermediate C2 (tert-butyl
3-methyl-N-(oxomethylene)-L-valinate) was prepared in a similar
fashion to Intermediate C1, substituting tert-butyl
3-methyl-L-valinate (Bachem AG) for methyl 3-methyl-L-valinate in
Step 3 of intermediate B1 of WO'566, p 14.
Preparation of Intermediate D1.
##STR00182## ##STR00183##
[0717] Steps 1 and 2. Preparation of trans-cyclopropanol mixture
D1-2 and D1-3: THF (1000 mL) was introduced in a three neck round
bottomed flask containing Mg (32.2 g, 1.34 mol). A solution of
7-bromohept-1-ene (216 g, 1.22 mol) in THF (600 mL) was introduced
to an addition funnel. One crystal of iodine and 20 mL of
7-bromohept-1-ene solution were added to the reaction. The solution
was heated to reflux, and the remainder of the 7-bromohept-1-ene
solution was added drop wise. After the addition was complete, the
mixture was refluxed for an additional 2 h then allowed to cool to
rt to produce a solution of Grignard reagent D1-1, which was then
added dropwise to a solution of ethyl formate (30 g, 0.41 mol) and
Ti(Oi-Pr).sub.4 (115.2 g, 0.41 mol) in THF (1200 mL) at rt. After
stirring overnight, the mixture was poured into 1600 mL of 10%
aqueous H.sub.2SO.sub.4 and extracted with MTBE (1500 mL three
times). The combined organic layers were washed with brine, dried
over MgSO.sub.4, filtered and concentrated in vacuo. The residue
was purified by silica gel chromatography to afford 31.0 g of a
mixture of trans-cyclopropyl alcohols D1-2 and D1-3 as a yellow
oil. .sup.1H NMR: (400 MHz, CDCl.sub.3): .delta. 5.77-5.70 (m, 1H),
4.96-4.86 (m, 2H), 3.15-3.12 (m, 1H), 2.03-1.98 (m, 2H), 1.75 (br
s, 1H), 1.45-1.37 (m, 2H), 1.20-1.15 (m, 1H), 1.06-1.01 (m, 1H),
0.89-0.82 (m, 1H), 0.63-0.59 (m, 1H), 0.24 (q, J=6.0 Hz, 1H). Step
3. Preparation of cyclopropyl acetate mixture D1-4 and D1-5: To a
1000 mL round bottom flask was added trans-cyclopropyl alcohol
mixture D1-2 and D1-3 (60.3 g, 0.48 mol), 700 mL of DCM and TEA
(62.9 g, 0.62 mol) prior to cooling the solution in an acetone/ice
bath to an internal temp of <5.degree. C. Acetyl chloride (41.3
g, 0.53 mol) was added dropwise to the solution over a 30 min
period while maintaining an internal temp<10.degree. C. The
resulting slurry was then warmed to rt and stirred for 2 h. The
reaction mixture was diluted with 350 mL of water. The biphasic
mixture was transferred to a separatory funnel and the aqueous
layer removed. The organic layer was washed with 480 mL of 2 N
aqueous HCl and then with 500 mL of sat. aqueous NaHCO.sub.3 prior
to drying over MgSO.sub.4. The solvent was removed in vacuo. The
residue was purified by silica gel chromatography to afford a
mixture D1-4 and D1-5 (56.3 g) as a yellow oil. TLC Information
(PE/EtOAc=5/1) R.sub.f (starting material)=0.4; R.sub.f
(product)=0.8. Step 4. Preparation of D1-3: To a 1000 mL
round-bottom flask was added a solution of mixture D1-4 and D1-5
(39 g, 0.23 mol) in 680 mL of MTBE saturated with aqueous 0.1 M pH
7 phosphate buffer. The flask was placed in an ice bath to maintain
an internal temperature of approximately 10.degree. C. throughout
the hydrolysis reaction which was initiated by the addition of 3.0
g of Novozyme 435. The reaction was aged at 10.degree. C. for
approximately 6 h until conversion had reached about 40%. The
reaction mixture was filtered, and the solid immobilized enzyme was
washed three times with 200 mL of MTBE. The resulting MTBE solution
was concentrated in vacuo. The residue was purified by silica gel
chromatography to afford D1-3 (11.3 g) as a yellow oil. .sup.1H
NMR: (400 MHz, CDCl.sub.3) .delta. 5.80-5.75 (m, 1H), 5.02-4.91 (m,
2H), 3.20-3.17 (m, 1H), 2.09-2.03 (m, 3H), 1.50-1.43 (m, 2H),
1.26-1.22 (m, 1H), 1.17-1.08 (m, 1H), 1.07-0.89 (m, 1H), 0.70-0.65
(m, 1H), 0.32-0.27 (m, 1H). Step 5. Preparation of D1-6:
Cyclopropanol D1-3 (17.7 g, 0.140 mol) was dissolved in 300 mL of
MeCN at 0.degree. C. To the solution was added DSC (72.0 g, 0.280
mol) and TEA (42.42 g, 0.420 mol). The reaction mixture was warmed
to 40.degree. C. and stirred overnight and then concentrated in
vacuo. The residue was purified by silica gel chromatography to
afford D1-6 (25.8 g) as a yellow solid. .sup.1H NMR: (400 MHz,
CDCl.sub.3) .delta. 5.84-5.77 (m, 1H), 5.05-4.96 (m, 2H), 4.09-4.03
(d, J=24 Hz, 1H), 2.86 (s, 4H), 2.12-2.06 (m, 2H), 1.58-1.51 (m,
2H), 1.33-1.27 (m, 3H), 1.09 (m, 1H), 0.68-0.62 (m, 1H). Step 6.
Preparation of D1-7: To a solution of D1-6 (10 g, 0.0374 mol) in
THF (374 mL) was added L-tert-leucine methyl ester hydrochloride
(10.2 g, 0.056 mol) and TEA (11.3 g, 0.112 mol). The solution was
stirred overnight at 40.degree. C. The mixture was concentrated in
vacuo. The residue was diluted with EtOAc and washed with water and
brine, dried over anhydrous sodium sulfate, filtered and
concentrated in vacuo. The residue was purified by silica gel
chromatography to afford D1-7 (10.2 g) as a yellow oil. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.16H.sub.28NO.sub.4: 298.2.
found: 298.0. Step 7. Preparation of Intermediate D1: A solution of
D1-7 (20 g, 0.067 mol) in 2:1 mixture of MeOH/H.sub.2O (447 mL/223
mL) was treated with LiOH*H.sub.2O (11.3 g, 0.269 mol) and then
heated at 60.degree. C. for 4 h. The reaction mixture was cooled,
concentrated to half volume and extracted with MTBE. Then the
aqueous solution was acidified with aqueous 1 N HCl (400 mL) and
extracted with EtOAc (400 mL.times.3), the combined organic layers
were washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated to afford Intermediate D1 (18 g). .sup.1H NMR: (400
MHz, CDCl.sub.3) .delta. 10.5-9.4 (br, 1H), 5.82-5.71 (m, 1H),
5.20-5.17 (m, 1H), 4.99-4.91 (m, 2H), 4.19-4.16 (m, 1H), 3.86-3.68
(m, 1H), 2.09-2.03 (m, 2H), 1.53-1.32 (m, 2H), 1.30-1.20 (m, 2H),
1.18-1.13 (m, 1H), 1.11-0.99 (s, 9H), 0.80-0.75 (m, 1H), 0.49-0.47
(m, 1H).
Preparation of Intermediate D2.
##STR00184##
[0718] Step 1. Preparation of Intermediate D2: To a suspension of
D1-6 (600 mg, 2.25 mmol) and (S)-2-amino-2-cyclopentylacetic acid
hydrochloride salt (386 mg, 2.7 mmol, Betapharma Inc.) in THF (20
mL) were added distilled water (6 mL) and triethylamine (0.94 mL,
6.74 mmol). The homogeneous solution was allowed to stir for
.about.18 h. The THF was evaporated and the aqueous residue was
diluted with water (20 mL). The mixture was basified with 1 N NaOH
(pH>10) and then washed twice (20 mL) with ethyl acetate. The
aqueous phase was then acidified with 1 N HCl (pH<2) and the
resulting solution was extracted twice (20 mL) with ethyl acetate.
The combined organic phase was dried over anhydrous MgSO.sub.4 and
concentrated to afford Intermediate D2 (500 mg) as a brown oil.
This was used without purification in a subsequent step. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.16H.sub.26NO.sub.4: 296.2.
found: 296.3.
Preparation of Intermediate D3.
##STR00185##
[0719] Step 1. Preparation of Intermediate D3: To a suspension of
D1-6 (800 mg, 3 mmol) and (S)-2-amino-2-cyclohexylacetic acid (519
mg, 3.3 mmol; Alfa Aesar) in water (15 mL) was added
K.sub.3PO.sub.4 (1.27 g, 6 mmol). The homogeneous solution was
allowed to stir at rt for 5 h. To the reaction mixture was charged
with water (15 mL) and EtOAc (15 mL). The organic layer was
separated, and the aqueous layer was extracted with EtOAc (10 mL).
Organic layers were combined, washed with 1 N HCl, H.sub.2O and
brine, and dried over Na.sub.2SO.sub.4. Concentration of the
organic solution afforded Intermediate D3 (850 mg) as an oil that
was used subsequently without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.17H.sub.28NO.sub.4: 310.4. found:
310.3.
Preparation of Intermediate D4.
##STR00186## ##STR00187##
[0720] Step 1. Preparation of D4-2: Bicyclic alcohol D4-1 (2.9 g,
29.5 mmol, prepared according to Section A, Intermediate 1 of U.S.
Pat. No. 8,178,491 B2 (hereinafter "US '491"), p 192.) was
dissolved in DCM (60 mL) and TEA (8.2 mL, 59 mmol) was added. The
stirred solution was cooled to 0.degree. C. and MsCl (3.4 mL, 44
mmol) was added. The reaction mixture was allowed to warm to rt
gradually. After 18 h, the reaction mixture was poured into
H.sub.2O. The aqueous layer was extracted 2.times. with DCM then
the combined organics were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. The crude material was
purified by silica gel chromatography (20% to 50% EtOAc/Hex) to
afford D4-2 (3.73 g). Step 2. Preparation of D4-3: NaH (1.69 g,
42.3 mmol) was suspended in 100 mL THF and the mixture was cooled
to 0.degree. C. Diethyl malonate (6.4 mL, 47 mmol) was added
dropwise over 4 min and the stirred mixture was warmed to rt. After
another hour, mesylate D4-2 (3.73 g, 21.2 mmol) in 20 mL THF was
added and the reaction mixture was heated to reflux for 15 h. After
this period, the reaction mixture was cooled to rt and poured into
saturated aqueous NaHCO.sub.3. The aqueous layer was extracted
2.times. with EtOAc. Then, the organics were dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The crude
material was purified by silica gel chromatography (0% to 15%
EtOAc/Hex) to afford D4-3 (4.64 g). Step 3. Preparation of D4-4:
Malonate D4-3 (4.64 g, 19.3 mmol) was dissolved in 20 mL DMSO then
NaCl (1.24 g, 21.2 mmol) and water (0.694 mL, 38.6 mmol) were
added. The stirred mixture was heated to 170.degree. C. for 48 h
then cooled to rt and diluted with Et.sub.2O. The organic solution
was washed with H.sub.2O twice, then brine, dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The crude
material was purified by silica gel chromatography (5% to 15%
EtOAc/Hex) to afford D4-4 (2.83 g). Steps 4 and 5. Preparation of
D4-5: A solution of ethyl ester D4-4 (2.83 g, 16.8 mmol) and LiOH
(1 M in H.sub.2O, 34 mL, 34 mmol) in EtOH (68 mL) was stirred at rt
o/n then concentrated under reduced pressure to remove EtOH. The
remaining material was diluted with H.sub.2O and washed twice with
DCM. The aqueous phase was acidified to pH 1-2 with 10% HCl and
then extracted three times with DCM. This DCM solution was dried
over MgSO.sub.4, filtered and concentrated under reduced pressure.
The resulting crude carboxylic acid was dissolved in DCM (100 mL)
and treated with DMF (5 drops). Oxalyl chloride (2.2 mL, 25 mmol)
was added carefully. After stirring o/n, the reaction mixture was
concentrated under reduced pressure to afford D4-5, which was
carried on without further purification. Step 6. Preparation of
D4-6: (S)-4-Benzyl-2-oxazolidinone (3.57 g, 20.2 mmol) was
dissolved in THF (80 mL) and cooled to -78.degree. C. n-BuLi (1.6 M
in hexane, 12.6 mL, 20.2 mmol) was added dropwise over 7 min and
the reaction mixture was allowed to stir at -78.degree. C. for 30
min. This solution, containing the lithiated oxazolidinone was then
added by cannula to a -78.degree. C. solution of acid chloride D4-5
(16.8 mmol) in THF (80 mL) over 6 min. After stirring at
-78.degree. C. for an additional 30 min, the reaction mixture was
quenched by addition of 1 M aqueous NaHSO.sub.4. The aqueous phase
was extracted with EtOAc and the organic layer was dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
crude material was purified by silica gel chromatography (10% to
40% EtOAc/Hex) to afford D4-6 (4.32 g). LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.18H.sub.22NO.sub.3: 300.16. found: 300.14. Step 7.
Preparation of D4-7: A solution of KHMDS (0.5 M in PhMe, 3.4 mL,
1.7 mmol) in THF (5 mL) was cooled to -78.degree. C. and a separate
-78.degree. C. solution of oxazolidinone D4-6 (465 mg, 1.55 mmol)
in THF (5 mL) was added dropwise by cannula. After 30 min, a
-78.degree. C. solution of trisyl azide (576 mg, 1.86 mmol) in THF
(5 mL) was added by cannula. Three min later, the reaction was
quenched by addition of AcOH (0.41 mL, 7.13 mmol) and the reaction
mixture was heated to 30.degree. C. for 2 h. After cooling, the
mixture was poured into brine. The aqueous layer was extracted
three times with DCM. The combined organics were dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
crude material was purified by silica gel chromatography (4% to 25%
EtOAc/Hex) to afford azide D4-7 (367 mg). LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.18H.sub.21N.sub.4O.sub.3: 341.16.
found: 341.10. Step 8. Preparation of D4-8: Azide D4-7 (367 mg,
1.08 mmol) and di-tert-butyl dicarbonate (471 mg, 2.16 mmol) were
dissolved in EtOAc (20 mL). 10% Pd/C (197 mg) was added and the
atmosphere replaced with H.sub.2. The suspension was stirred under
1 atm H.sub.2 for 20 h, then filtered over Celite and concentrated
under reduced pressure. The crude residue was purified by silica
gel chromatography (15% to 30% EtOAc/Hex) to afford D4-8 (376 mg).
LCMS-ESI (m/z): [M-(t-Bu)+H].sup.+ calcd for
C.sub.19H.sub.23N.sub.2O.sub.5: 359.16. found: 359.43.
[0721] Steps 9 and 10. Preparation of D4-9: Carbamate D4-8 (376 mg,
0.907 mmol) was dissolved in THF (9 mL) and cooled to 0.degree. C.
H.sub.2O.sub.2(30% in H.sub.2O, 0.463 mL, 4.54 mmol) and LiOH (1M
in H.sub.2O, 2.7 mL, 2.7 mmol) were added. The reaction was allowed
to stir at 0.degree. C. for another 2 h and was then concentrated
under reduced pressure. The resulting concentrate was poured into
H.sub.2O and the aqueous solution was washed twice with Et.sub.2O,
then acidified to pH 1-2 and extracted three times with DCM. The
combined extracts were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. The resulting crude residue
was dissolved in DCM (8 mL) and MeOH (1 mL) and treated with
trimethylsilyldiazomethane (2 M in hexane, 0.9 mL, 1.8 mmol). After
stirring for 40 min at rt, the reaction was quenched by addition of
10% AcOH/MeOH and concentrated under reduced pressure. The residue
was purified by silica gel chromatography (4% to 25% EtOAc/Hex) to
afford D4-9 (167 mg). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
4.98 (d, J=7.8 Hz, 1H), 4.22 (t, J=7.0 Hz, 1H), 3.70 (s, 3H), 1.89
(m, 1H), 1.77-1.46 (m, 4H), 1.42 (s, 9H), 1.22 (m, 2H), 0.28 (dd,
J=7.2 Hz, 13.3 Hz, 1H), 0.13 (d, J=3.7 Hz, 1H).
Step 11. Preparation of D4-10: Carbamate D4-9 (223 mg, 0.828 mmol)
was dissolved in DCM (4 mL) and treated with HCl (4.0 M in dioxane,
1 mL, 4.0 mmol). After stirring at rt for 17 h, the reaction
mixture was concentrated under reduced pressure to afford amine
hydrochloride salt D4-10, which was carried on without
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.9H.sub.16NO.sub.2: 170.12. found: 170.04. Steps 12 and 13.
Preparation of Intermediate D4: Amine hydrochloride salt D4-10
(0.828 mmol, theoretical) in H.sub.2O (1.4 mL) was treated with a
freshly prepared solution of D1-6 (1.35 mmol) in DMF (1.4 mL).
K.sub.3PO.sub.4 (703 mg, 3.31 mmol) was added and the reaction
mixture was stirred for 2 h at rt. After dilution with EtOAc, the
organic layer was washed with 10% aqueous HCl and brine, then dried
over MgSO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by silica gel chromatography (0% to 25%
EtOAc/Hex) to afford the expected carbamate (239 mg). LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.18H.sub.28NO.sub.4: 322.20.
found: 323.00. This material (239 mg, 0.744 mmol) was dissolved in
MeOH and treated with LiOH (1.0 M in H.sub.2O, 5.0 mL, 5.0 mmol).
After stirring at rt for 1 h, the MeOH was removed under reduced
pressure. The aqueous solution was acidified to pH 1-2 with 10%
aqueous HCl and was extracted three times with DCM. The combined
organics were dried over MgSO.sub.4, filtered and concentrated
under reduced pressure to afford Intermediate D4 (229 mg).
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.17H.sub.26NO.sub.4: 308.2. found: 307.9.
Preparation of Intermediate D5.
##STR00188##
[0722] Intermediate D5 was prepared according to the procedure
detailed in Li, H., et al. Synlett 2011, 10, 1454. Preparation of
Intermediate mixture D6.
##STR00189##
Step 1. Preparation of diastereomeric carbamate mixture D6-1:
Intermediate C2 (1.34 g, 6.31 mmol),
(.+-.)-trans-1-methyl-2-(pent-4-enyl)cyclopropanol (590 mg, 4.208
mmol; prepared according to procedure for Intermediate C3,
WO2011014487, p. 36), DMAP (514 mg, 4.21 mmol), and DIPEA (2.93 mL,
16.83 mmol) were combined in toluene (14 mL). The reaction was
heated at 90.degree. C. for 18 h. The reaction was diluted with
Et.sub.2O (25 mL) and 1 N aqueous HCl (75 mL), stirred well, and
organics were removed. The aqueous layer was extracted three times
with ether (50 mL), the organics were combined, washed with brine,
dried over MgSO.sub.4, filtered, and concentrated to give a crude
oil, which was purified via silica gel chromatography to give 1:1
diastereomeric mixture D6-1 as a clear oil (820 mg). LCMS-ESI.sup.+
(m/z): [M+Na].sup.+ calcd for C.sub.20H.sub.35NNaO.sub.4: 376.3.
found: 376.2. Step 2: Preparation of diastereomeric Intermediate
mixture D6. The diastereomeric mixture D6-1 was taken up in DCM (2
mL) and treated with TFA (2 mL) at room temperature. After 1.5 h,
the reaction was concentrated in vacuo and co-evaporated with
chloroform repeatedly to remove residual TFA and purified via
silica gel chromatography to give 1:1 diastereomeric mixture of
Intermediate D6 as a brown oil, (536 mg). LCMS-ESI.sup.+ (m/z):
[M+Na].sup.+ calcd for C.sub.16H.sub.27NNaO.sub.4: 320.2. found:
320.1.
Preparation of Intermediate D7.
##STR00190##
[0723] Step 1. Preparation of D7-1:
(1R,2R)-1-methyl-2-(pent-4-enyl)cyclopentanol (220.9 mg, 1.313
mmol; prepared according to procedure for Intermediate B26,
International Patent Publication No. WO 2008/057209 (hereinafter
"WO '209"), p. 45) and Intermediate C1 (337.1 mg, 1.969 mmol) were
treated with DIPEA (0.91 mL, 5.252 mmol) and DMAP (160.4 mg, 1.313
mmol) in toluene (4.4 mL). The mixture was heated at 85.degree. C.
for 21 h. The solution was diluted with ether (80 mL). The solution
was washed with 1 N aqueous HCl (30 mL) and brine (30 mL)
successively. Obtained organic layer was dried over
Na.sub.2SO.sub.4. After removal of drying agent by a filtration,
the solvent was removed under reduced pressure. The residue was
purified by silica gel chromatography (13% ethyl acetate in
hexanes) to give D7-1 (249.5 mg, 0.735 mmol) as colorless oil.
.sup.1H NMR (300 MHz, CDCl.sub.3, rotamers expressed as total H
value.times.fraction present) .delta. 5.76-5.92 (m, 1H), 5.12 (d,
J=9.6 Hz, 1H), 5.02 (d, J=16.8 Hz, 1H), 4.96 (d, J=9.6 Hz, 1H),
4.13 (d, J=9.6 Hz, 1H), 3.81 (s, 3.times.4/10H), 3.73 (s,
3.times.6/10H), 1.80-2.15 (m, 7H), 1.04-1.74 (m, 6H), 1.36 (s, 3H),
1.04 (s, 9.times.4/10H), 0.97 (s, 9.times.6/10H). Step 2.
Preparation of Intermediate D7: Ester D7-1 (249.5 mg, 0.735 mmol)
was treated with 2 M aqueous LiOH aqueous solution (2 mL, 4.0 mmol)
in MeOH/THF (4 mL/4 mL) at rt for 25 h. The reaction mixture was
then treated with 1 N aqueous HCl (5 mL) and aqueous brine (25 mL)
to slightly acidify. The mixture was extracted three times with
CH.sub.2Cl.sub.2 (30 mL). The organic layer was washed with aqueous
brine (30 mL). Obtained organic layer was dried over
Na.sub.2SO.sub.4. After removal of drying agent by filtration, the
solvent was removed under a reduced pressure to give Intermediate
D7 (191.2 mg, 0.587 mmol) as a colorless oil which was used
subsequently without further purification. .sup.1H NMR (300 MHz,
CDCl.sub.3) b 9.00 (br s, 1H), 5.72-5.90 (m, 1H), 5.12 (d, J=9.6
Hz, 1H), 5.00 (d, J=16.8 Hz, 1H), 4.94 (d, J=9.6 Hz, 1H), 4.13 (d,
J=9.6 Hz, 1H), 1.80-2.16 (m, 7H), 1.04-1.74 (m, 6H), 1.35 (s, 3H),
1.02 (s, 9H). Preparation of Intermediate mixture D8.
##STR00191##
Step 1. Preparation of D8-2: To a solution of intermediate D8-1
(500 mg, 3.24 mmol, prepared according to WO '209, p. 36) in DCM
(6.65 mL) was added Dess-Martin periodinane (1.37 g, 3.24 mmol) at
rt under an argon atmosphere. After 6 h, the reaction mixture was
filtered through a pad of Celite and was directly purified by
silica gel chromatography (0-100% ethyl acetate/hexanes gradient)
to afford ketone D8-2 (252 mg) as a colorless oil. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 5.81 (ddt, J=16.9, 10.2, 6.6 Hz, 1H),
5.05-4.92 (m, 2H), 2.38-1.93 (m, 7H), 1.87-1.68 (m, 2H), 1.60-1.37
(m, 3H), 1.35-1.20 (m, 1H). Step 2. Preparation of diastereomeric
mixture D8-3: To a solution of ketone D8-2 (385 mg, 2.53 mmol) and
TMSCF.sub.3 (749 .mu.L, 5.07 mmol) in THF (2.3 mL) was added CsF
(7.0 mg, 46 .mu.mol) at rt under an argon atmosphere. After 2.5 h,
the reaction mixture was diluted with water (10 mL) and the
resulting mixture was extracted twice with DCM (10 mL). The
combined organic layers were dried over anhydrous sodium sulfate,
and were concentrated in vacuo. The crude residue was purified by
silica gel chromatography (0-100% ethyl acetate/hexanes gradient)
to afford silyl ether D8-3 (714 mg, 1:1 diastereomeric mixture) as
a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.67
(ddt, J=13.3, 10.1, 6.7 Hz, 1H), 4.91-4.76 (m, 2H), 2.02-1.00 (m,
13H), 0.00 (s, 9H). Step 3. Preparation of diastereomeric mixture
D8-4: To a solution of D8-3 (700 mg, 2.38 mmol) in THF (11.9 mL)
was added TBAF (1M in THF, 2.38 mL, 2.38 mmol) at rt under an argon
atmosphere. After 30 min, the reaction mixture was diluted with
dichloromethane (100 mL). The resulting mixture was washed with
saturated aqueous sodium bicarbonate solution (75 mL), was dried
over anhydrous sodium sulfate, and was concentrated in vacuo. The
crude residue was purified by silica gel chromatography (0-100%
ethyl acetate/hexanes gradient) to afford alcohol D8-4 (418 mg, 1:1
diastereomeric mixture) as a colorless oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 5.81 (dt, J=16.8, 6.6 Hz, 1H), 5.09-4.88 (m,
2H), 2.20-1.91 (m, 4H), 1.86-1.08 (m, 10H). Step 4. Preparation of
diastereomeric mixture D8-5: A solution of D8-4 (380 mg, 1.72
mmol), Intermediate C1 (295.7 mg, 1.72 mmol), DIPEA (1.20 mL, 6.88
mmol), and DMAP (210 mg, 1.72 mmol) in toluene (8.6 mL) was heated
to 85.degree. C. under an argon atmosphere. After 20 h, the
reaction mixture was allowed to cool to rt and was diluted with
ethyl acetate (100 mL). The resulting mixture was washed with 1 N
HCl solution (50 mL), saturated aqueous sodium bicarbonate solution
(50 mL), and brine (50 mL). The organic layer was dried over
anhydrous sodium sulfate, and was concentrated in vacuo. The crude
residue was purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford carbamate D8-5 (550 mg, 1:1
diastereomeric mixture) as a colorless oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 5.81 (ddt, J=16.7, 9.8, 6.6 Hz, 1H), 5.37 (d,
J=9.4 Hz, 1H), 5.06-4.89 (m, 2H), 4.16-4.07 (m, 1H), 3.75 (s, 3H),
2.84-2.29 (m, 2H), 2.27-1.89 (m, 3H), 1.85-1.12 (m, 8H), 0.98 (s,
9H). Step 5. Preparation of diastereomeric Intermediate mixture D8:
To a solution of carbamate D8-5 (500 mg, 1.27 mmol) in DCE (6.4 mL)
was added trimethyltin hydroxide (2.30 g, 12.7 mmol) at rt under an
argon atmosphere, and the resulting mixture was heated to
65.degree. C. After 21 h, the reaction mixture allowed to cool to
rt and was diluted with 1 N HCl solution (50 mL). The resulting
mixture was extracted with ethyl acetate (2.times.50 mL). The
combined organic extracts were dried over anhydrous sodium sulfate
and were concentrated in vacuo to afford Intermediate D8 (575 mg,
1:1 diastereomeric mixture) as a colorless oil, which was used
subsequently without further purification. .sup.1H NMR (400 MHz,
CDCl.sub.3) b 5.90-5.71 (m, 1H), 5.32 (d, J=9.3 Hz, 1H), 5.07-4.89
(m, 2H), 4.16 (d, J=9.8 Hz, 1H), 2.83-2.30 (m, 2H), 2.27-1.87 (m,
3H), 1.83-1.12 (m, 8H), 1.04 (s, 9H). Preparation of Intermediate
mixture D9 and D10.
##STR00192##
Steps 1 and 2: Preparation of racemate D9-1: Magnesium metal (1.32
g, 54.3 mmol) was added to a 2-neck flask fitted with a reflux
condenser and the vessel was flushed with Ar. THF (42 mL) was added
followed by iodine (ca. 5 mg). The stirred suspension was heated to
45.degree. C. and 5-bromopent-1-ene was added (1.2 g, 8.1 mmol) in
one portion. After stirring several minutes, additional
5-bromopent-1-ene (5.5 g, 37 mmol) was added at a rate sufficient
to maintain gentle reflux. The resulting mixture was stirred at
50.degree. C. for 15 min and was then cooled to ambient temperature
and used immediately in the following step. A suspension of CuI
(630 mg, 3.3 mmol) in THF (24 mL) under Ar was cooled to -5 OC. An
aliquot of pent-4-enylmagnesium bromide (ca. 0.95 M, 20 mL, 19
mmol) prepared in step 1 was added over 5 min, and the resulting
mixture was stirred for an additional 15 min. The reaction mixture
was then cooled to -20.degree. C., and
(.+-.)-exo-2,3-epoxynorbornane (1.5 g, 14 mmol) was added as a
solution in THF (5 mL) over 1 min. Two additional portions of THF
(2.5 mL each) were used to ensure complete transfer, and the
resulting mixture was stirred for 20 min. The reaction was then
removed from the cold bath and warmed to rt. After stirring an
additional 1.75 h, the reaction was quenched with saturated aqueous
NH.sub.4Cl (5 mL) and was filtered with EtOAc (100 mL) and H.sub.2O
(100 mL) through Celite. The phases were separated, and the organic
phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated
to afford (.+-.)-D9-1 as a colorless residue (813 mg). .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 5.90-5.67 (m, 1H), 5.04-4.86 (m, 2H),
3.12 (s, 1H), 2.20-1.92 (m, 5H), 1.69-1.57 (m, 1H), 1.55-1.12 (m,
9H), 1.03-0.84 (m, 1H). Step 3. Preparation of diastereomeric
Intermediate mixture D9 and D10: Alcohol mixture (.+-.)-D9-1 (813
mg, 4.51 mmol) was dissolved in DMF (4.5 mL). Pyridine (370 .mu.L,
4.5 mmol) was added followed by DSC (1.5 g, 5.8 mmol). The reaction
mixture was heated to 45.degree. C. and was stirred for 4 h. The
reaction mixture was then cooled to 0.degree. C. and water (4.5 mL)
was added dropwise over 2 min. The reaction mixture was stirred for
5 min and was removed from the cold bath. After an additional 5
min, the reaction mixture was cooled to 0.degree. C. and
L-tert-leucine (835 mg, 6.37 mmol) and K.sub.3PO.sub.4 (2.70 g,
12.7 mmol) were added. The mixture was stirred for 10 min and was
removed from the cold bath. After stirring an additional 24 h, the
mixture was diluted with EtOAc (30 mL), acidified with 1 M aqueous
HCl (15 mL), and diluted with 0.2 M aqueous HCl (15 mL). The phases
were separated, and the organic phase was washed with 0.2 M aqueous
HCl (2.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated to afford diastereomeric Intermediate mixture D9 and
D10 (1.64 g). LCMS-ESI.sup.- (m/z): [M-H].sup.- calcd for
C.sub.19H.sub.30NO.sub.4: 336.2. found: 336.0.
Preparation of Intermediate D11.
##STR00193##
[0724] Step 1. Preparation of 011-1: To a mixture of 01 (1.0 g,
3.53 mmol), sodium periodate (2.26 g, 10.59 mmol) in 24 mL THF and
12 mL water was added Os EnCat.TM. 40 (0.25 mmol/g loading, 282 mg,
0.071 mmol, Sigma-Aldrich). The mixture was stirred for 3 days.
Water (50 mL) was added and the mixture was filtered. The filter
cake was washed with water (total volume 400 mL) and ethyl acetate
(total volume 600 mL). The filtrate layers were separated. The
organic phase was dried over sodium sulfate, filtered and
concentrated to give 011-1 (1.56 g) which was used without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.14H.sub.24NO.sub.5: 286.2 found: 286.1. Step 2. Preparation
of D11-2: To a solution of 011-1 (3.05 g, 10.7 mmol) in MeOH (50
mL) at 0.degree. C. was added sodium borohydride in portions (809
mg, 21.4 mmol). The reaction mixture was stirred at rt for 6 h. The
mixture was diluted with 50 mL ethyl acetate and 50 mL brine and
the layers were separated. The organic phase was extracted with two
25 mL portions of ethyl acetate. The combined organic phase was
dried over sodium sulfate, filtered and concentrated. The crude
product mixture was purified by silica gel chromatography (EtOAc in
hexanes: 10% to 100%) to give 011-2 (380 mg). LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.14H.sub.26NO.sub.5: 288.2. found:
288.1. Step 3. Preparation of Intermediate D11: To a solution of
D11-2 (283 mg, 0.98 mmol) in THF (2.8 mL) at 0.degree. C. was added
1-nitro-2-selenocyanatobenzene (336 mg, 1.47 mmol) and
tributylphosphine (363 .mu.L, 1.47 mmol). The cooling bath was
removed and the mixture was stirred for 25 minutes at rt. The
reaction was again cooled to 0.degree. C. and was treated with 30%
hydrogen peroxide solution (0.665 mL, 5.85 mmol) and stirred for 1
h at rt and then heated at 60.degree. C. for 1 h. The reaction was
diluted with EtOAc and the desired product was extracted into
aqueous sodium bicarbonate. The bicarbonate extract was acidified
with 2 N HCl and extracted with ethyl acetate. The organic phase
was dried over sodium sulfate and concentrated to give Intermediate
D11 (136 mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.14H.sub.24NO.sub.4: 270.2. found: 270.1. Preparation of
Intermediate mixture D12 and D13.
##STR00194##
Step 1: Preparation of D12-1: To a solution of
K.sub.2Cr.sub.2O.sub.7(121 g, 0.41 mol) in H.sub.2O (1.5 L) was
added dropwise H.sub.2SO.sub.4 (143 g, 1.46 mol) at rt and the
mixture was stirred for 1 h. The mixture was then cooled to
0.degree. C. and D4-1 (80 g, 0.814 mol; prepared according to
Section A, Intermediate 1 of US '491, p 192.) in MTBE (1.5 L) was
added dropwise. The reaction mixture was stirred at rt for 2 h. The
aqueous phase was extracted with MTBE (3.times.500 mL), dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The crude product
was purified by distillation (20 mmHg, bp: 60-62.degree. C.) to
provide D12-1 as a pale yellow liquid (60 g). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 2.57-2.63 (m, 2H), 2.14-2.19 (d, J=20 Hz, 2H),
1.52-1.57 (m, 2H), 0.89-0.94 (m, 1H), -0.05--0.02 (m, 1H). Step 2:
Preparation of (.+-.)-D12-2: Under Ar, a mixture of THF (4.4 mL)
and HMPA (1.8 mL) was cooled to -78.degree. C. A 1 M solution of
LiHMDS in THF (2.2 mL, 2.2 mmol) was added. Ketone D12-1 (202 mg,
2.10 mmol) was added as a solution in THF (2 mL) over 1 min,
washing with additional THF (2.times.1 mL) to ensure complete
transfer. After 25 min, 5-iodopent-1-ene (prepared according to
Jin, J. et. al. J. Org. Chem. 2007, 72, 5098-5103) (880 mg, 4.5
mmol) was added over 30 s by syringe. After 10 min, the reaction
was placed in a cold bath at -45.degree. C. and was warmed to
-30.degree. C. over 1.5 h. The reaction was quenched with saturated
aqueous NH.sub.4Cl (15 mL) and was diluted with EtOAc (30 mL) and
H.sub.2O (15 mL). The phases were separated, and the aqueous phase
was extracted with EtOAc (30 mL). The combined organics were dried
over Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue that was purified by silica gel chromatography (0% to 15%
EtOAc in hexanes) to provide (+/-)-D12-2 a colorless oil (162 mg).
.sup.1H NMR (400 MHz, CDCl.sub.3) 5.82-5.67 (m, 1H), 5.03-4.87 (m,
2H), 2.61-2.51 (m, 1H), 2.11 (d, J=19.1 Hz, 1H), 2.08-1.99 (m, 3H),
1.61-1.40 (m, 5H), 1.36-1.28 (m, 1H), 0.92-0.81 (m, 1H),
-0.03--0.11 (m, 1H). Step 3: Preparation of (.+-.)-D12-3 and
(.+-.)-D12-4: A solution of (.+-.)-D12-2 (142 mg, 0.865 mmol) in
THF (4 mL) was cooled to -78.degree. C. A 1 M THF solution of
LiBHEt.sub.3 (1.3 mL, 1.3 mmol) was added dropwise over 30 s. The
reaction was stirred 15 min and was removed from the cold bath.
After warming to rt (15 min), the reaction was quenched with
saturated aqueous NH.sub.4Cl (1 mL). The resulting mixture was
diluted with Et.sub.2O (20 mL) and H.sub.2O (20 mL). The phases
were separated, and the aqueous phase was extracted with Et.sub.2O
(20 mL). The combined organics were dried over MgSO.sub.4,
filtered, and concentrated to a crude residue. Purification by
silica gel chromatography (0% to 10% EtOAc in hexanes) provided 133
mg of a mixture of diastereomers (.+-.)-D12-3 and (.+-.)-D12-4. The
combined material from two experiments (253 mg) was further
purified by silica gel chromatography (0% to 15% EtOAc in hexanes)
to provide (.+-.)-D12-3 (150 mg) and (.+-.)-D12-4 (58 mg) as
colorless oils. .sup.1H NMR for (.+-.)-D12-3 (300 MHz, CDCl.sub.3)
.delta. 5.91-5.69 (m, 1H), 5.07-4.88 (m, 2H), 3.97 (d, J=6.7 Hz,
1H), 2.19-1.99 (m, 3H), 1.84-1.73 (m, 1H), 1.62 (d, J=14.1 Hz, 1H),
1.54-1.40 (m, 2H), 1.32-1.17 (m, 3H), 1.16-1.06 (m, 1H), 0.60-0.43
(m, 2H). .sup.1H NMR for (.+-.)-D12-4 (300 MHz, CDCl.sub.3) .delta.
5.95-5.73 (m, 1H), 5.09-4.88 (m, 2H), 4.05-3.86 (m, 1H), 2.17-1.84
(m, 4H), 1.72-1.34 (m, 5H), 1.28-1.08 (m, 3H), 0.49-0.36 (m, 1H),
0.21-0.11 (m, 1H). Step 4: Preparation of diastereomeric
Intermediate mixture D12 and D13: A mixture of (.+-.)-D12-3 (150
mg, 0.90 mmol) was dissolved in DMF (1.0 mL). Pyridine (75 .mu.L,
0.92 mmol) and DSC (302 mg, 1.18 mmol) were added, and the reaction
was stirred at 45.degree. C. for 21.5 h. The reaction was then
placed in an ice water bath and H.sub.2O (1.0 mL) was added
dropwise via syringe over 1 min. The mixture was removed from the
cold bath and allowed to stir 5 min. The mixture was re-cooled in
an ice water bath and L-tert-leucine (154 mg, 1.17 mmol) was added
followed by K.sub.3PO.sub.4 (502 mg, 2.36 mmol). The reaction
mixture was removed from the cold bath and allowed to stir at rt
for 24 h. The mixture was then diluted with EtOAc (40 mL) and 1 M
aqueous HCl (20 mL). The phases were separated, and the aqueous
phase was extracted with EtOAc (30 mL). The combined organic phase
was washed with 0.2 M aqueous HCl (2.times.20 mL), dried over
MgSO.sub.4, filtered, and concentrated to afford diastereomeric
Intermediate mixture D12 and D13 (300 mg) as a colorless oil.
LCMS-ESI.sup.- (m/z): [M-H].sup.- calcd for
C.sub.18H.sub.28NO.sub.4: 322.2. found: 322.0).
Preparation of Intermediate D12.
##STR00195##
[0725] Step 1: Preparation of D12-5: To a solution of
(1S,4R)-cis-4-acetoxy-2-cyclopent-1-ol (Aldrich, 10 g, 70.4 mmol),
triethylamine (48.8 mL, 350 mmol), and DMAP (4.29 g, 35.2 mmol) in
dichloromethane (352 mL) was added pivaloyl chloride (10.8 mL,
87.75 mmol) dropwise via syringe at 0.degree. C. under an argon
atmosphere. After 2 h, the reaction mixture was diluted with
saturated aqueous sodium bicarbonate solution (500 mL), and
extracted with dichloromethane (2.times.500 mL). The combined
organic extracts were dried over anhydrous sodium sulfate and were
concentrated in vacuo to afford D12-5 (15.0 g) as a colorless oil.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.08 (br s, 2H), 5.54
(td, J=8.0, 4.1 Hz, 2H), 2.88 (dt, J=14.9, 7.5 Hz, 1H), 2.07 (s,
3H), 1.69 (dt, J=14.7, 4.1 Hz, 1H), 1.20 (s, 9H). Step 2:
Preparation of D12-6: To a solution of D12-5 (15.0 g, 70.4 mmol) in
methanol (352 mL) was added potassium carbonate (9.73 g, 70.4 mmol)
at rt under an argon atmosphere. After 5 h, the reaction mixture
was filtered and was concentrated in vacuo. The residue was
dissolved into ethyl acetate (500 mL) and the resulting mixture was
washed with water (500 mL) and brine (500 mL), dried over anhydrous
sodium sulfate, and concentrated in vacuo. The crude residue was
purified by silica gel chromatography (0-100% ethyl
acetate/hexanes) to afford D12-6 (12.0 g) as a colorless oil.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.11 (br d, J=5.5 Hz,
1H), 5.97 (br d, J=5.6 Hz, 1H), 5.48 (br s, 1H), 4.73 (br s, 1H),
2.82 (dt, J=14.6, 7.3 Hz, 1H), 1.67 (s, 1H), 1.61 (dt, J=14.5, 4.0
Hz, 1H), 1.20 (s, J=3.8 Hz, 9H). Step 3: Preparation of D12-7: To a
solution of copper(I) cyanide (5.10 g, 57.0 mmol) in diethyl ether
(95 mL) was added pent-4-enylmagnesium bromide (Novel Chemical
Solutions, 0.5 M in THF, 114 mL, 57.0 mmol) dropwise via cannula
over a 30 min period at 0.degree. C. under an argon atmosphere.
After 10 min, a solution of D12-6 (3.50 g, 19.0 mmol) in diethyl
ether (10 mL) was added slowly via cannula. The reaction mixture
was then allowed to slowly warm to rt. After 16 h, the resulting
mixture was quenched with saturated aqueous ammonium chloride
solution (400 mL) and the resulting mixture was extracted into
ethyl acetate (2.times.400 mL). The combined organic phases were
washed with brine (400 mL), dried over anhydrous sodium sulfate,
and concentrated in vacuo. The crude residue was purified by silica
gel chromatography (0-100% ethyl acetate/hexanes) to afford D12-7
(2.4 g) as a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 5.80 (ddt, J=16.9, 10.2, 6.7 Hz, 1H), 5.69 (dd, J=5.8, 1.7
Hz, 1H), 5.65 (d, J=7.2 Hz, 1H), 5.00 (dd, J=17.1, 1.3 Hz, 1H),
4.94 (d, J=10.2 Hz, 1H), 4.12-4.05 (m, 1H), 2.69 (ddd, J=17.2, 6.4,
1.5 Hz, 1H), 2.54-2.45 (m, 1H), 2.24 (d, J=17.2 Hz, 1H), 1.69 (br
s, 1H), 1.52-1.19 (m, 6H). Step 4: Preparation of
(1S,2R,3R,5S)-D12-3: To a solution of D12-7 (20 mg, 0.13 mmol), and
diethyl zinc (1 M in hexanes, 132 .mu.L, 0.132 mmol) in diethyl
ether (0.66 mL) was added diiodomethane (21 .mu.L, 0.26 mmol) at rt
under an argon atmosphere. After 2 h, the reaction mixture was
quenched with 1 N aqueous HCl solution (0.66 mL). After 5 min, the
resulting yellow mixture was diluted with saturated aqueous sodium
bicarbonate solution (5 mL) and the resulting mixture was extracted
with dichloromethane (3.times.5 mL). The combined organic extracts
were dried over anhydrous sodium sulfate solution, and were
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes) to afford
(1S,2R,3R,5S)-D12-3 (10 mg) as a colorless oil. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 5.83 (ddt, J=16.9, 10.2, 6.7 Hz, 1H), 5.02
(d, J=17.2 Hz, 1H), 4.96 (d, J=11.3 Hz, 1H), 4.00 (d, J=6.7 Hz,
1H), 2.19-2.02 (m, 3H), 1.82 (t, J=7.2 Hz, 1H), 1.64 (d, J=14.2 Hz,
1H), 1.55-1.42 (m, 2H), 1.38-1.20 (m, 4H), 1.19-1.08 (m, 1H),
0.62-0.47 (m, 2H). Step 5: Preparation of Intermediate D12: Alcohol
(1S,2R,3R,5S)-D12-3 (0.450 g, 2.7 mmol) was taken up in DMF (2.7
mL) and treated subsequently with DSC (0.92 g, 3.52 mmol) and
pyridine (0.22 mL, 2.8 mmol). The reaction was then heated to
50.degree. C. o/n. The reaction was then cooled to 0.degree. C. and
water (5.5 mL) was added dropwise over 1 min. The resulting opaque
suspension was stirred at rt for 10 min before recooling to
0.degree. C. The reaction was then treated subsequently with
L-tert-leucine (0.462 g, 3.5 mmol) and K.sub.3PO.sub.4 (1.5 g, 7.0
mmol) and allowed to warm to rt overnight with vigorous stirring.
The resulting opaque suspension was diluted with EtOAc and 1 M
aqueous HCl. Additional HCl (12 M) was added dropwise to adjust the
pH 3. The aqueous layer was extracted with EtOAc and the combined
organics were washed with brine and dried over anhydrous
MgSO.sub.4. Following concentration in vacuo, Intermediate D12 was
obtained (1.72 g) as a viscous, colorless oil that is contaminated
with small amounts of DMF and EtOAc. The material was used in
subsequent reactions without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.18H.sub.30NO.sub.4: 324.2. found
324.7.
Preparation of Intermediate D14.
##STR00196##
[0726] Step 1. Preparation of Intermediate D14. Carbonate D1-6 (862
mg, 3.23 mmol) was treated with
(S)-2-amino-2-(1-methylcyclopentyl)acetic acid hydrochloride (750
mg, 3.87 mmol; prepared according to Robl, J. A., et al. J. Med.
Chem., 2004, 47, 2587), THF (28 mL), H.sub.2O (8.4 mL) and TEA (1.4
mL, 9.7 mmol). The reaction mixture was stirred for 16 h and the
THF was removed in vacuo. The remaining material was diluted with
H.sub.2O and the pH adjusted to .about.10-12 by addition of 10%
aqueous NaOH. The aqueous phase was washed twice with EtOAc and
then acidified to pH 1-2 with 10% aqueous HCl. The acidic solution
was extracted 3.times. with EtOAc. The combined extractions were
dried over anhydrous MgSO.sub.4, filtered and concentrated in
vacuo. The initial EtOAc washings (of the basic aqueous solution)
were washed with 10% aqueous HCl, dried over MgSO.sub.4, filtered
and concentrated in vacuo. The combined concentrates were purified
by silica gel chromatography (50% to 100% EtOAc/Hex) to afford
Intermediate D14 (980 mg). LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd
for C.sub.17H.sub.28NO.sub.4: 310.2. found 310.0. Preparation of
Intermediate mixture D15.
##STR00197##
Step 1. Preparation of (.+-.)-D15-1: To a solution of titanium(IV)
isopropoxide (11.3 g, 40.0 mmol) in THF (160 mL) was added methyl
magnesium bromide (3 M in Et.sub.2O, 20 mL, 60.0 mmol) dropwise via
syringe at rt under an argon atmosphere. After 10 min, the reaction
mixture was cooled to 0.degree. C. and a solution of methyl
propionate (3.80 mL, 40.0 mmol) in THF (10 mL) was added slowly via
syringe. After 5 min, hept-6-enylmagnesium bromide (Novel Chemical
Solutions, 0.5 M in THF, 160 mL, 80 mmol) was added dropwise via
addition funnel over 1 h. After 2.5 h, the reaction mixture was
quenched with 10% aqueous sulfuric acid (100 mL) and the resulting
mixture was extracted with diethyl ether (2.times.200 mL). The
organic phase was dried over anhydrous sodium sulfate and was
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes) to afford
(.+-.)-D15-1 (3.03 g, 50%) as a colorless oil. .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 5.77 (ddt, J=16.9, 10.2, 6.7 Hz, 1H),
5.03-4.86 (m, 2H), 2.04 (q, J=6.1 Hz, 2H), 1.75-1.14 (m, 6H), 1.04
(t, J=7.4 Hz, 3H), 1.01-0.91 (m, 1H), 0.89-0.71 (m, 2H), 0.02 (t,
J=5.5 Hz, 1H). Step 2. Preparation of diastereomeric Intermediate
mixture D15: Racemic alcohol mixture (.+-.)-D15-1 (2.00 g, 13.0
mmol) was dissolved in DMF (13.0 mL). Pyridine (1.05 mL, 13.0 mmol)
was added followed by DSC (4.00 g, 15.6 mmol). The reaction mixture
was heated to 50.degree. C. and was stirred for 20 h. The reaction
mixture was then cooled to rt and water (13 mL) was added dropwise
over 2 min. L-tert-leucine (2.17 g, 13.0 mmol) and K.sub.3PO.sub.4
(8.28 g, 39.0 mmol) were then added and the reaction mixture was
warmed to 50.degree. C. After 5 h, the reaction mixture was allowed
to cool to rt and was diluted with water (500 mL). The resulting
mixture was washed with dichloromethane (100 mL). The aqueous phase
was then acidified to pH 2 with 2 N aqueous HCl solution, and was
extracted with DCM (2.times.400 mL). The combined organic extracts
were dried over anhydrous sodium sulfate and were concentrated
under reduced pressure to afford diastereomeric Intermediate
mixture D15 (4.5 g) as a pale orange oil, which was used
subsequently without further purification.
Preparation of Intermediate D16:
##STR00198##
[0728] Intermediate D16 was prepared in a similar fashion to the
preparation of Intermediate D12, substituting but-3-enylmagnesium
bromide for pent-4-enylmagnesium bromide in Step 3. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for C.sub.17H.sub.28NO.sub.4: 310.2. found
310.8.
Preparation of Intermediate D17:
##STR00199##
[0729] Step 1. Preparation of intermediate mixture D17.
(.+-.)-trans-1-methyl-2-(but-3-enyl)cyclopropanol (900 mg, 0.13
mmol), prepared according to procedure for Intermediate B2,
International Patent Publication No. WO 2012/40040 (hereinafter "WO
'040"), p. 38, was dissolved in DMF (6 mL). Pyridine (577 .mu.L,
7.13 mmol) was added followed by DSC (2.37 g, 9.27 mmol). The
reaction mixture was heated to 40.degree. C. and was stirred for 18
h. The reaction mixture was then cooled to 0.degree. C. and water
(6 mL) was added dropwise over 5 min. The reaction mixture was
stirred for 5 min and was removed from the cold bath. After an
additional 5 min, the reaction mixture was cooled to 0.degree. C.
and L-tert-leucine (1.21 g, 9.27 mmol) and K.sub.3PO.sub.4 (4.69 g,
22.1 mmol) were added. The mixture was stirred for 10 min and was
removed from the cold bath. After stirring an additional 6 h, the
mixture was diluted with EtOAc (30 mL), acidified with 1 M aqueous
HCl (25 mL), and diluted with 0.2 M aqueous HCl (25 mL). The phases
were separated, and the organic phase was washed with 0.2 M aqueous
HCl (2.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated to afford diastereomeric carbamate mixture D17 (2.10
g). LCMS-ESI.sup.+ (m/z): [M+Na].sup.+ calcd for
C.sub.15H.sub.25NNaO.sub.4: 306.2. found: 306.1.
Preparation of Intermediate D18:
##STR00200##
[0731] Step 1. Preparation of D18-1: (Prepared according to
WO2011013141) To a solution of (S)-4-amino-2-hydroxybutanoic acid
(15 g, 126 mmol) in methanol (95 mL) was added concentrated
sulfuric acid (8 mL), and the reaction was heated to reflux. After
18 h, the resulting mixture was allowed to cool to room temperature
and was concentrated in vacuo. The residue was slurried with ethyl
acetate (95 mL) and D18-1 was collected by vacuum filtration.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.69 (br s, 1H), 4.31
(ddd, J=9.2, 8.1, 2.2 Hz, 1H), 3.49 (d, J=5.6 Hz, 1H), 3.41 (tt,
J=9.2, 1.7 Hz, 1H), 3.33 (td, J=9.4, 6.5 Hz, 1H), 2.81 (br s, 1H),
2.59-2.48 (m, 1H), 2.09 (dq, J=12.9, 9.1 Hz, 1H).
[0732] Step 2. Preparation of D18-2: To a solution of D18-1 (4.5 g,
44 mmol), 4-nitrobenzoic acid (8.19 g, 49 mmol), and
triphenylphosphine (22.4 g, 132 mmol) in tetrahydrofuran (220 mL)
was added diisopropyl azodicarboxylate (12.1 mL, 61.6 mmol)
dropwise via syringe at 23.degree. C. under an argon atmosphere.
After 20 h, the resulting cloudy orange reaction mixture was
concentrated in vacuo and methanol (200 mL) followed by potassium
carbonate (15 g, 109 mmol) were added and the reaction was stirred
at 23.degree. C. After an additional 5 h, the resulting mixture was
diluted with chloroform (200 mL) and was filtered. The filtrate was
concentrated in vacuo and the crude residue was taken up into water
(150 mL) and 1N aqueous hydrochloric acid solution (50 mL). The
aqueous layer was washed with ethyl acetate (3.times.200 mL) to
remove organic by-products, and was concentrated in vacuo to crude
afford D18-2 that was used directly in the next step. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 4.28 (t, J=8.4 Hz, 1H), 3.43-3.20 (m,
1H), 2.56-2.39 (m, 1H), 1.96 (dq, J=12.7, 8.7 Hz, 1H).
[0733] Step 3. Preparation of D18-3: To a solution of crude D18-2
(5 g, 49.5 mmol) and imidazole (3.4 g, 49.5 mmol) in DMF (247 mL)
was added TBSCl (7.5 g, 49.5 mmol) at 0.degree. C. under an argon
atmosphere. The resulting mixture was allowed to warm to 23.degree.
C. After 7 h, additional imidazole (7 g, 102 mmol) and TBSCl (16 g,
106 mmol) were added sequentially. After an additional 16 h, the
resulting mixture was diluted with 1N aqueous hydrochloric acid
solution (1 L) and was extracted with ethyl acetate (1 L). The
organic layer was split and was washed with brine (1 L), was dried
with anhydrous sodium sulfate, and was concentrate in vacuo. The
crude residue was purified by silica gel chromatography (0-100%
ethyl acetate/hexanes gradient) to afford D18-3. .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 5.99 (s, 1H), 4.26 (t, J=7.7 Hz, 1H),
3.44-3.33 (m, 1H), 3.30-3.19 (m, 1H), 2.45-2.29 (m, 1H), 2.11-1.95
(m, 1H), 0.91 (s, 9H), 0.02 (s, 3H), 0.00 (s, 3H).
[0734] Step 4. Preparation of D18-4: To a solution of D18-3 (1.00
g, 4.65 mmol), DMAP (57.8 mg, 0.465 mmol), and triethylamine (1.29
mL, 9.3 mmol) in dichloromethane (23.3 mL) was added di-tert-butyl
dicarbonate (1.5 g, 6.97 mmol) at 23.degree. C. under and argon
atmosphere. After 20 h, the reaction mixture was purified directly
by silica gel chromatography (0-100% ethyl acetate/hexanes
gradient) to afford D18-4. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 4.31 (dd, J=9.4, 7.9 Hz, 1H), 3.79 (ddd, J=11.0, 8.9, 2.2
Hz, 1H), 3.53-3.41 (m, 1H), 2.34-2.21 (m, 1H), 1.92 (dq, J=12.2,
9.2 Hz, 1H), 1.53 (s, 9H), 0.91 (s, 9H), 0.17 (s, 3H), 0.13 (s,
3H).
[0735] Step 5. Preparation of D18-5: To a solution of D18-4 (700
mg, 2.22 mmol) in tetrahydrofuran (11.1 mL) was added
pent-4-enylmagnesium bromide (Novel Chemical Solutions, 0.5 M in
2-MeTHF, 4.89 mL, 2.44 mmol) at -78.degree. C. dropwise via syringe
under an argon atmosphere. After 1 h, the reaction mixture was
quench with saturated aqueous ammonium chloride solution (50 mL)
and was allowed to warm to room temperature. The resulting mixture
was extracted with ethyl acetate (2.times.100 mL), and the combined
organic extracts were washed with brine (100 mL), were dried over
anhydrous sodium sulfate and were concentrated in vacuo. The crude
residue was purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford D18-5. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 5.77-5.62 (m, 1H), 4.95 (d, J=15.8 Hz, 1H),
4.92 (d, J=10.2 Hz, 1H), 4.26 (app t, J=8.4 Hz, 1H), 3.77-3.69 (m,
1H), 3.41 (td, J=10.4, 6.7 Hz, 1H), 2.48 (t, J=7.4 Hz, 2H),
2.28-2.17 (m, 1H), 1.91-1.78 (m, 2H), 1.77-1.65 (m, 1H), 1.60
(quin, J=7.3 Hz, 2H), 1.47 (s, 9H), 0.85 (s, 9H), 0.11 (s, 3H),
0.07 (s, 3H).
[0736] Step 6. Preparation of D18-6: To a solution of D18-5 (740
mg, 1.92 mmol) and triethylsilane (6.10 mL, 38.4 mmol) in
dichloromethane (9.6 mL) was added boron trifluoride diethyl
etherate (308 .mu.L, 2.50 mmol) at -78.degree. C. dropwise via
syringe under an argon atmosphere. After 1 h, the reaction mixture
was allowed to warm to room temperature. After an additional 4 h,
the reaction was quenched with saturated aqueous ammonium chloride
solution (10 mL), and was diluted with saturated sodium bicarbonate
solution (50 mL). The resulting mixture was extracted with ethyl
acetate (50 mL), and the organic layer was dried over anhydrous
sodium sulfate and was concentrated in vacuo to afford crude free
amine which was used directly in the next step. To a solution of
the crude free amine, and triethylamine (535 .mu.L, 3.84 mmol) in
tetrahydrofuran (9.6 mL) was added acetic anhydride (146.5 .mu.L,
1.55 mmol) at room temperature under an argon atmosphere. After 1
h, the resulting mixture was concentrated in vacuo and the crude
residue was purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford D18-6 (2:1 diastereomeric
mixture favoring desired
1-((2S,3R)-3-(tert-butyldimethylsilyloxy)-2-(pent-4-enyl)pyrrolidin-1-yl)-
ethanone diastereomer). .sup.1H NMR (400 MHz, CDCl.sub.3, Minor
diastereomer denoted by *) .delta. 5.80-5.64 (m, 1H, 1H*),
5.01-4.82 (m, 2H, 2H*), 4.10 (d, J=4.2 Hz, 1H*), 4.04 (d, J=3.7 Hz,
1H), 3.82 (dd, J=10.3, 4.0 Hz, 1H), 3.66-3.56 (m, 1H*), 3.55-3.29
(m, 2H, 1H*), 3.24-3.16 (m, 1H*), 2.37-2.25 (m, 1H*), 2.08-1.88 (m,
2H, 1H*), 2.03 (s, 3H*), 2.00 (s, 3H), 1.81-1.61 (m, 2H, 2H*),
1.50-1.01 (m, 4H, 4H*), 0.85 (s, 9H*), 0.80 (s, 9H), 0.10 (s, 3H*),
0.09 (s, 3H*), 0.00 (br s, 6H).
[0737] Step 7. Preparation of D18-7: To a solution of D18-6 (338
mg, 1.08 mmol) in tetrahydrofuran (21 mL) was added TBAF (1M in
tetrahydrofuran, 21 mL, 21 mmol) at 0.degree. C. under an argon
atmosphere. After 17 h, the reaction mixture was concentrated in
vacuo and was directly purified by silica gel chromatography
(0-100% ethyl acetate/hexanes gradient) to afford D18-7 (102 mg,
2:1 diastereomeric mixture favoring desired 1
1-((2S,3R)-3-hydroxy-2-(pent-4-enyl)pyrrolidin-1-yl)ethanone
diastereomer). .sup.1H NMR (400 MHz, CDCl.sub.3 Minor diastereomer
denoted by *) .delta. 5.84-5.70 (m, 1H, 1H*), 5.06-4.91 (m, 2H,
2H*), 4.25 (d, J=3.7 Hz, 1H*), 4.20 (d, J=3.7 Hz, 1H), 3.98 (dd,
J=9.2, 4.2 Hz, 1H), 3.76-3.68 (m, 1H*), 3.67-3.59 (m, 1H, 1H*),
3.55-3.46 (m, 1H, 2H*), 3.02-2.94 (m, 1H), 2.22-1.85 (m, 2H, 2H*),
2.10 (s, 3H*), 2.07 (s, 3H), 1.82-1.59 (m, 2H, 2H*), 1.55-1.13 (m,
4H, 4H*).
[0738] Step 8. Preparation of D18-8: To a solution of D18-7 (102
mg, 0.518 mmol) and pyridine (8 .mu.L, 0.104 mmol) was added DSC
(159.2 mg, 0.621 mmol) at room temperature, and the resulting
mixture was heated to 45.degree. C. After 16 h, the reaction
mixture was allowed to cool to room temperature and water (518
.mu.L), L-tert-leucine (86.5 mg, 0.518 mmol), and K.sub.3PO.sub.4
(330 mg, 1.55 mmol) were sequentially added, and the resulting
mixture was heated to 50.degree. C. After 6 h, the reaction mixture
was allowed to cool to room temperature and was diluted with 1N
aqueous hydrochloric acid solution (10 mL). The resulting mixture
was extracted with dichloromethane (2.times.10 mL), and the
combined organic extracts were dried over anhydrous sodium sulfate
and were concentrated in vacuo to afford D18-8 (2:1 diastereomeric
mixture favoring the desired
(S)-2-(((2S,3R)-1-acetyl-2-(pent-4-enyl)pyrrolidin-3-yloxy)carbonylamino)-
-3,3-dimethylbutanoic acid). .sup.1H NMR (400 MHz, CDCl.sub.3,
Minor diastereomer denoted by *) .delta. 5.85-5.65 (m, 1H, 1H*),
5.39 (d, J=9.3 Hz, 1H*), 5.34 (d, J=9.2 Hz, 1H), 5.07-4.87 (m, 3H,
3H*), 4.16-4.03 (m, 1H, 1H*), 3.83-3.45 (m, 3H, 3H*), 2.30-1.95 (m,
8H), 2.30-1.95 (m, 2H, 3H*), 1.82-1.65 (m, 2H, 1H*), 2.11 (s, 3H),
2.09 (s, 3H*), 1.58-1.13 (m, 4H, 4H*), 1.01 (br s, 9H, 9H*).
Preparation of Intermediate mixture D19.
##STR00201##
[0739] Steps 1 and 2: Preparation of D19-1: A 1.0 M THF solution of
KHMDS (10 mL, 10 mmol) was diluted with THF (10 mL) under Ar and
the resulting solution was cooled to -78.degree. C. in a
CO.sub.2:acetone bath. Bicyclo[3.1.1]heptan-2-one (1.0 g, 9.1 mmol,
see: Yin, et. al. J. Org. Chem. 1985, 50, 531) was added as a
solution in THF (5 mL) over 2 min, washing with additional THF
(2.times.2.5 mL) to ensure complete transfer. The resulting mixture
was stirred for 30 min, and
N-(5-Chloro-2-pyridyl)bis(trifluoromethanesulfonimide) (3.8 g, 9.7
mmol) was added as a solution in THF (10 mL) over 2 min, washing
with additional THF (2.times.2.5 mL). The resulting mixture was
stirred for 5 min and removed from the cold bath. After stirring an
additional 30 min, the reaction was diluted with Et.sub.2O (70 mL)
and 1 M aqueous HCl (50 mL). The phases were separated, and the
organic phase was washed with 1 M aqueous NaOH (2.times.30 mL). The
combined organic phase was dried over MgSO.sub.4, filtered and
concentrated to afford a crude residue. This was filtered through a
plug of silica with 30% EtOAc in hexanes to afford a crude residue
of (1.24 g) that was used directly in the following step. Step 2:
To a solution of 3-butenal diethyl acetal (1.4 mL, 8.3 mmol) under
Ar cooled in an ice water bath was added a 0.5 M THF solution of
9-Borabicyclo[3.3.1]nonane (15.9 mL, 7.95 mmol) over 3 min. The
reaction was stirred for 20 h, with the cold bath being allowed to
expire overnight. A 3 M aqueous solution of NaOH (2.9 mL, 8.7 mmol)
was then added, and, after stirring 20 min, the resulting solution
was transferred in its entirety to a flask containing the product
from Step 1 (ca. 5.16 mmol) and PdCl.sub.2(dppf).CH.sub.2Cl.sub.2
(420 mg, 0.51 mmol). The resulting mixture was heated to 60.degree.
C. After stirring 14 h, the reaction mixture was diluted with
Et.sub.2O (50 mL) and H.sub.2O (50 mL). The phases were separated,
and the organic phase was dried over MgSO.sub.4, filtered, and
concentrated. Purification by silica gel chromatography (0% to 10%
EtOAc in hexanes following pre-equilibration with 1% Et.sub.3N in
EtOAc) provided intermediate D19-1. .sup.1H NMR (300 MHz,
CDCl.sub.3) 5.36-5.28 (m, 1H), 4.59 (t, J=5.6 Hz, 1H), 3.73-3.58
(m, 2H), 3.54-3.39 (m, 2H), 2.72-2.60 (m, 1H), 2.45-2.34 (m, 3H),
2.23-2.08 (m, 4H), 1.89-1.76 (m, 2H), 1.67 (dt, J=16.1, 6.9 Hz,
2H), 1.58-1.47 (m, 2H), 1.23 (t, J=7.0 Hz, 6H).
[0740] Step 3: Preparation of D19-2: A solution of olefin D19-1
(660 mg, 2.77 mmol) in THF (25 mL) was cooled in an ice water bath.
BH.sub.3.Me.sub.2S was then added as a 1 M solution in
CH.sub.2Cl.sub.2 (2.9 mL, 2.9 mmol) over 1 min. The resulting
solution was stirred for 2 h in the ice water bath and was then
allowed to warm to r.t. After stirring an additional 3 h, the
reaction mixture was re-cooled in an ice water bath and was diluted
with 2 M aqueous NaOH (7 mL) followed by 30% aqueous
H.sub.2O.sub.2(7 mL). The resulting mixture was stirred an
additional 16 h as the cold bath was allowed to gradually expire.
The mixture was partitioned between Et.sub.2O (100 mL) and H.sub.2O
(50 mL), the phases were separated, and the organic phase was
washed with 0.5 M aqueous NaOH (50 mL). The organic phase was dried
over MgSO.sub.4, filtered, and concentrated to afford a crude
residue that was purified by silica gel chromatography (15% to 40%
EtOAc in hexanes) to afford 570 mg of Intermediate D19-2. .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 4.60 (t, J=5.6 Hz, 1H), 3.76-3.60
(m, 3H), 3.58-3.42 (m, 2H), 2.39-2.05 (m, 4H), 1.91-1.48 (m, 9H),
1.43-1.35 (m, 1H), 1.25 (t, J=7.0 Hz, 6H), 1.06-0.98 (m, 1H).
[0741] Steps 4 and 5: Preparation of D19-3: Acetal D19-2 (360 mg,
1.4 mmol) was dissolved in THF (8 mL) and H.sub.2O (2 mL).
para-Toluenesulfonic acid monohydrate (40 mg, 0.2 mmol) was added
and the resulting solution was stirred 16 h at r.t. The reaction
was diluted with Et.sub.2O (50 mL) and H.sub.2O (30 mL) and the
phases were separated. The aqueous phase was extracted with
Et.sub.2O (30 mL) and the combined organic phase was washed with
saturated aqueous NaHCO.sub.3 (15 mL). The organic phase was dried
over MgSO.sub.4, filtered, and concentrated to afford a crude
residue that was used immediately in the following step. Step 5:
Methyl triphenylphosphonium bromide (1.66 g, 4.6 mmol) was
suspended in THF (40 mL) under Ar and was cooled via a
CO.sub.2/acetone bath to -78.degree. C. A 1 M solution of NaHMDS in
THF (4.2 mL, 4.2 mmol) was added in dropwise fashion and the
resulting yellow suspension was stirred for 5 min. The mixture was
removed from the cold bath and stirring continued an additional 30
min. The mixture was then re-cooled to -78.degree. C. and the crude
residue from the previous step (ca. 1.4 mmol) was added as a
solution in THF (5 mL) over 5 min, washing with additional THF
(2.times.2.5 mL) to ensure complete transfer. The resulting mixture
was stirred for 5 min and was then placed in an ice water bath and
stirred an additional 1 h. The reaction was quenched with saturated
aqueous NH.sub.4Cl (20 mL) and was diluted with Et.sub.2O (30 mL)
and H.sub.2O (20 mL). The phases were separated and the organic
phase was dried over MgSO.sub.4, filtered, and concentrated onto 5
g silica gel. Purification by silica gel chromatography (10% to 30%
EtOAc in hexanes) provided D19-3. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 6.01-5.81 (m, 1H), 5.22-5.05 (m, 2H), 3.79-3.66 (m, 1H),
2.43-2.25 (m, 2H), 2.24-2.04 (m, 4H), 1.83-1.16 (m, 10H).
[0742] Step 6: Intermediate D19-3 (270 mg, 1.5 mmol) was dissolved
in DMF (2.0 mL). Pyridine (125 .mu.L, 1.5 mmol) and DSC (500 mg,
1.9 mmol) were added, and the reaction was stirred at 45.degree. C.
for 15 h. The reaction was then placed in an ice water bath and
H.sub.2O (2.0 mL) was added dropwise over 30 s. The mixture was
removed from the cold bath and allowed to stir 10 min. The mixture
was re-cooled in an ice water bath and L-tert-leucine (259 mg, 1.97
mmol) was added followed by K.sub.3PO.sub.4 (835 mg, 3.93 mmol).
The reaction mixture was removed from the cold bath and allowed to
stir at r.t. for 5.25 h. The mixture was then diluted with EtOAc
(40 mL), 1 M aqueous HCl (20 mL), and H.sub.2O (15 mL). The phases
were separated, and the aqueous phase was extracted with EtOAc (30
mL). The combined organic phase was washed with 0.2 M aqueous HCl
(2.times.25 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated to afford a mixture of diastereomers D19 (505 mg) as a
colorless oil. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.19H.sub.32NO.sub.4: 338.2. found: 337.8.
Preparation of Intermediate E1.
##STR00202##
[0743] Intermediate E1
(2-chloro-6-methoxy-3-(methylsulfonyl)quinoxaline) was prepared
according to Mahata, P. K., et al. Org. Lett. 2005, 7, 2169.
Preparation of Intermediate E2.
##STR00203##
[0744] Step 1. Preparation of E2-1: In a round bottom flask,
3-(benzyloxy)aniline (4.025 g, 20.20 mmol) and
1,1-bis(methylthio)-2-nitroethylene (3.338 g, 20.20 mmol) in
ethanol (40 mL) was refluxed for 24 h with constant stirring. The
reaction mixture was then cooled in an ice bath and diluted with
ether (150 mL). The mixture was filtered and washed with ether to
afford E2-1 (3.32 g) as a yellow solid which was used directly in
the following in step. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.16H.sub.17N.sub.2O.sub.3S: 317.1. found: 317.1. Step 2.
Preparation of E2-2: To a suspension of E2-1 (3.32 g, 10.49 mmol)
in 25 mL MeCN, POCl.sub.3 (2.93 mL, 31.5 mmol) was added dropwise
over 15 min with constant stirring. The reaction mixture was warmed
to 80.degree. C. and stirred for 5 h. The reaction was then cooled
to ambient temperature and neutralized with ice cold saturated
aqueous NaHCO.sub.3 solution, extracted three times with
CH.sub.2Cl.sub.2 (100 mL), washed with water, brine and dried over
anhydrous Na.sub.2SO.sub.4. The solvent was removed under reduced
pressure. The crude material was eluted through a plug of silica
with CH.sub.2Cl.sub.2. The solvent was removed under reduced
pressure and the solid was washed with MeCN to afford E2-2 (1.56 g)
as an off white solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.16H.sub.14ClN.sub.2OS: 317.1. found: 317.3. Step 3.
Preparation of Intermediate E2. A solution of mCPBA (1.87 g, 10.83
mmol) in CH.sub.2Cl.sub.2 (40 mL) was added dropwise to a stirred
solution of E2-2 (1.56 g, 4.92 mmol) in CH.sub.2Cl.sub.2 (40 mL) at
0.degree. C. over a period of 30 min. The reaction mixture was
further stirred at ambient temperature for 5 h. It was then poured
into ice could saturated aqueous NaHCO.sub.3 and partitioned with
CH.sub.2Cl.sub.2. The organic layer was then washed subsequently
with water, brine and dried over anhydrous Na.sub.2SO.sub.4. The
solvent was removed under reduced pressure and the crude material
was purified by normal phase chromatography with CH.sub.2Cl.sub.2
to provide the title compound Intermediate E2 as a pale yellow
solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.16H.sub.14ClN.sub.2O.sub.3S: 349.0. found: 349.0.
Preparation of Intermediate E3.
##STR00204##
[0745] Step 1. Preparation of E3-1: To a solution of
3-bromo-3,3-difluoroprop-1-ene (25.0 g, 159 mmol) and diethyl
oxalate (21.6 mL, 159 mmol) in THF (380 mL), diethyl ether (90 mL)
and n-pentane (90 mL) at -100.degree. C. was added dropwise
n-butyllithium (2.5 M in hexane, 67 mL, 167.6 mmol) over 30 min.
The reaction mixture was stirred at -95.degree. C. for 1 h and
-78.degree. C. for 2 h, and quenched with aq. NH.sub.4Cl (11 g in
150 mL of water). The mixture was extracted with ether (three
times). The organic layers were washed with 1 N aqueous HCl, brine,
and dried over Na.sub.2SO.sub.4, and concentrated to give the crude
residue, which was purified by silica gel chromatography (EtOAc in
hexanes: 0% to 40%) to give E3-1 (7.0 g). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 5.98-6.18 (m, 1H), 5.78 (dd, J=0.9 Hz, 13 Hz,
1H), 5.60 (dd, J=0.9 Hz, 11 Hz, 1H), 4.38 (q, J=6.9 Hz, 2H), 1.37
(t, J=7.2 Hz, 3H). Step 2. Preparation of E3-2 and E3-3: To a
solution of E3-1 (14.0 g, 78.6 mmol) and
4-methoxybenzene-1,2-diamine dihydrochloride (15.08 g, 71.4 mmol)
in EtOH (360 mL) at rt was added triethylamine (19.9 mL, 142.8
mmol). The reaction mixture was stirred at rt overnight. The
mixture was concentrated. Slurrying in dichloromethane (30 mL) and
filtering gave some separation of regioisomers with E3-2 as the
precipitating species. (16.5 g total yield from filtration and
subsequent chromatography). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 11.940 (br s, 1H), 7.850 (d, J=9 Hz, 1H), 6.985 (dd, J=3
Hz, 9 Hz, 1H), 6.754 (d, J=2 Hz, 1H), 6.625-6.498 (m, 1H), 5.907
(dt, J=17, 2 Hz, 1H), 5.601 (d, J=11 Hz, 1H), 3.938 (s, 3H). The
mixture was slurried, filtered, and concentrated once more, then
was purified by silica gel chromatography (EtOAc in hexanes: 5% to
34%) to give E3-3 (2.07 g) as the first eluting component. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 12.05 (br s, 1H), 7.850 (d, J=9
Hz, 1H), 6.986 (dd, J=3 Hz, 9 Hz, 1H), 6.761 (d, J=3 Hz, 1H),
6.597-6.526 (m, 1H), 5.91 (dt, J=17, 2 Hz, 1H), 5.601 (d, J=11 Hz,
1H), 3.939 (s, 3H). Step 3. Preparation of Intermediate E3: A
solution of E3-3 (2.07 g, 8.2 mmol in 1 mL DMF was treated with
POCl.sub.3 (0.8 mL) and heated at 65.degree. C. for 2.5 h. The
reaction was diluted with EtOAc and quenched by pouring into ice
water. The organic phase was washed subsequently with saturated
aqueous sodium bicarbonate and brine, dried over sodium sulfate and
concentrated to give 2.1 g of Intermediate E3. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 8.028 (d, J=10 Hz, 1H), 7.46 (dd, J=3 Hz,
9 Hz, 1H), 7.32 (d, J=3 Hz, 1H), 6.549-6.478 (m, 1H), 5.86 (dt,
J=17, 2 Hz, 1H), 5.67 (d, J=11 Hz, 1H), 3.981 (s, 3H).
Preparation of Intermediate E4.
##STR00205##
[0746] Intermediate E4 (2-chloro-3-(1,1-difluoroallyl)quinoxaline)
was prepared in a similar fashion to Intermediate E3, substituting
1,2-diaminobenzene for 4-methoxybenzene-1,2-diamine dihydrochloride
in Step 2.
Preparation of Intermediate E5.
##STR00206##
[0747] Intermediate E5 (2,6-dichloro-3-(methylsulfonyl)quinoxaline)
was prepared according to Mahata, P. K., et al. Org. Lett. 2005, 7,
2169.
Preparation of Intermediate E6.
##STR00207##
[0748] Step 1. Preparation of E6-1: A 1-L 3-necked round-bottom
flask was charged with a solution of 3-bromo-3,3-difluoroprop-1-ene
(25 g, 159.3 mmol) in DMF (360 mL) and water (90 mL). The resulting
solution was treated with ethyl 2-oxoacetate (33 mL, 1 M in
toluene), and In (25 g). The reaction mixture was stirred overnight
at rt and then extracted with 3.times.300 mL of ether. The organic
layers were combined, washed with 1.times.100 mL of saturated
aqueous NH.sub.4Cl and 1.times.100 mL of brine, dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo to afford E6-1
that was used subsequently without additional purification. Step 2.
Preparation of E6-2. To hydroxyester E6-1 (58.1 g, 323 mmol) was
added DCM (700 mL) in a 2 L 3-neck flask equipped with overhead
stirring and an internal temperature probe. Then TEMPO (5.4 g, 35
mmol), buffer solution (prepared by dissolving 4.2 g NaHCO.sub.3
and 0.53 g Na.sub.2CO.sub.3 per 100 mL water, 700 mL, 7v), and
NaOCl (Clorox 6.15% wt, 422 mL, 395 mmol) were sequentially added
to the flask at 20.degree. C. After 2 h the organic layer was
separated and the aqueous phase extracted with ethyl acetate
(2.times.300 mL). The combined organic layers were dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo to afford
E6-2. .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta. 5.98-6.18 (m, 1H),
5.78 (dd, J=0.9 Hz, 13 Hz, 1H), 5.60 (dd, J=0.9 Hz, 11 Hz, 1H),
4.38 (q, J=6.9 Hz, 2H), 1.37 (t, J=7.2 Hz, 3H). Step 3. Preparation
of E6-3. To a solution of ethyl
3,3-difluoro-2,2-dihydroxypent-4-enoate E6-2 (57.4 g, 292 mmol) in
THF (725 mL) and water (131 mL) was added LiOH*H.sub.2O (22 g, 529
mmol) at 20.degree. C. After 2.5 h, the reaction mixture was
concentrated in vacuo. The solid residue was suspended in water
(300 mL) and the resulting mixture was acidified to pH=1 with
concentrated aqueous hydrochloric acid solution. The resulting
mixture was stirred until all solids were dissolved (.about.1.5 h),
and then sodium chloride was added until the solution was
saturated. The resulting solution was extracted with MTBE
(2.times.500 mL) and ethyl acetate (2.times.500 mL), and the
combined organic layers were dried over anhydrous Na.sub.2SO.sub.4
and were concentrated in vacuo. The crude orange solid residue was
suspended into DCM (100 mL) and was stirred until the solids were
finely distributed before hexanes (75 mL) were slowly added via
addition funnel. The resulting solids were collected by vacuum
filtration through a medium fritted funnel and washed with 1:1
dichloromethane/hexanes (2.times.10 mL) to afford the desired
product. .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 13.17 (bs,
1H), 6.18-6.01 (m, 1H), 5.64-5.52 (m, 2H). Step 4. Preparation of
E6-4 and E6-5: A solution of E6-3 (0.5 g, 3.3 mmol) in EtOH (12 mL)
was treated with 3,4-diaminobenzonitrile (0.47 g, 3.5 mmol). The
reaction mixture was heated at 80.degree. C. for 1 h, then
concentrated in vacuo. The resulting residue was absorbed on silica
gel, then was purified by column chromatography to give E6-4 (0.5
g) as the first eluting component. .sup.1H-NMR (400 MHz,
CD.sub.3OD) .delta. 8.01 (d, 1H), 7.65 (dd, 2H), 6.49 (m, 1H), 5.80
(dt, 1H), 5.60 (d, 1H). E6-5 (0.2 g) was recovered as the second
eluting component. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta. 8.25
(d, 1H), 7.87 (dd, 1H), 7.41 (d, 1H), 6.49 (m, 1H), 5.80 (dt, 1H),
5.59 (d, 1H). Step 5. Preparation of Intermediate E6: A solution of
E6-4 (0.5 g, 2 mmol in 4.5 mL DMF was treated with POCl.sub.3 (3
mL) and heated at 65.degree. C. for 3 h. The reaction was diluted
with EtOAc and quenched by pouring into ice water. The organic
phase was washed subsequently with saturated aqueous NaHCO.sub.3
and brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated
in vacuo to give 0.48 g of Intermediate E6
(3-chloro-2-(1,1-difluoroallyl)quinoxaline-6-carbonitrile).
.sup.1H-NMR (400 MHz, CD.sub.3OD) .delta. 8.52 (s, 1H), 8.30 (d,
1H), 8.13 (dd, 1H), 6.55 (m, 1H), 5.84 (dt, 1H), 5.72 (d, 1H).
Preparation of Intermediate E7
##STR00208##
[0749] Step 1. Preparation of E7-1: To a solution of E3-1 (1.84 g,
10.93 mmol) and 4-(difluoromethoxy)benzene-1,2-diamine (1.90 g,
10.93 mmol, prepared according to Reference Example 30y of
WO2003035065, p. 511.) in DMF (40 mL) at rt was added DIPEA (9.5
mL, 54.65 mmol) and HATU (6.23 g, 16.4 mmol). The reaction mixture
was stirred at room temperature for 24 h, diluted with ethyl
acetate (100 mL), washed with water (100 mL) and brine (50 mL). The
mixture was concentrated in vacuo. Purification via silica gel
chromatography (EtOAc in hexanes: 20% to 60%) provided E7-1 (800
mg) as the later eluting fraction of two with the similar mass
spectra. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.12H.sub.9F.sub.4N.sub.2O: 289.2. found: 289.0. Step 2:
Preparation of Intermediate E7: Hydroxyquinoxaline E7-1 (800 mg,
2.8 mmol), POCl.sub.3 (1.65 mL, 3.0 mmol) and DMF (10 mL) are
combined at rt and then heated to 65.degree. C. for 2.5 h at which
time additional POCl.sub.3 (0.2 mL, 0.36 mmol) was added. The
reaction was heated an additional 3 h at 65.degree. C. then cooled
to rt. The reaction was quenched by addition of cold water (30 mL),
and taken up into ethyl acetate (50 mL), washed with saturated
aqueous Na.sub.2CO.sub.3 (100 mL) followed by brine (50 mL), and
dried over anhydrous MgSO.sub.4. The resulting solution was
concentrated in vacuo to give Intermediate E7 (859 mg) which was
used subsequently without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.12H.sub.8ClF.sub.4N.sub.2O: 307.0.
found: 307.0.
Preparation of Intermediate E8.
##STR00209##
[0750] Intermediate E8
(2-chloro-6-fluoro-3-(methylsulfonyl)quinoxaline) was prepared
according to Mahata, P. K., et al. Org. Lett. 2005, 7, 2169.
Preparation of Intermediate E9.
##STR00210##
[0751] 2,7-dichloro-3-(prop-2-en-1-yl)quinazolin-4(3H)-one
(Intermediate E9) was prepared according to Step 3 of Intermediate
D5 of WO '040 p 53-4.
PREPARATION OF EXAMPLES
Example 1
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-3,6--
dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanoc-
yclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8--
carboxamide
##STR00211## ##STR00212## ##STR00213##
[0753] Step 1. Preparation of 1-1: A mixture containing
Intermediate B4 (2.03 g, 6.44 mmol), Intermediate E1 (1.6 g, 5.85
mmol), and cesium carbonate (3.15 g, 9.66 mmol) in MeCN (40 mL) was
stirred vigorously at rt under an atmosphere of Ar for 16 h. The
reaction was then filtered through a pad of Celite and the filtrate
concentrated in vacuo. The crude material was purified by silica
gel chromatography to provide 1-1 as a white solid (2.5 g).
LCMS-ESI.sup.+ (m/z): [M-Boc+2H].sup.+ calcd for
C.sub.20H.sub.27ClN.sub.3O.sub.4: 408.9. found: 408.6.
[0754] Step 2. Preparation of 1-2: To a solution 1-1 (2.5 g, 4.92
mmol) in dioxane (10 mL) was added hydrochloric acid in dioxane (4
M, 25 mL, 98.4 mmol) and the reaction stirred at rt for 5 h. The
crude reaction was concentrated in vacuo to give 1-2 as a white
solid (2.49 g) that was used in subsequently without further
purification. LCMS-ESI.sup.+ (m/z): [M].sup.+ calcd for
C.sub.20H.sub.26ClN.sub.3O.sub.4: 407.9. found: 407.9.
[0755] Step 3. Preparation of 1-3: To a DMF (35 mL) solution of 1-2
(2.49 g, 5.61 mmol), Intermediate D1 (1.75 mg, 6.17 mmol) and DIPEA
(3.9 mL, 22.44 mmol) was added COMU (3.12 g, 7.29 mmol) and the
reaction was stirred at rt for 3 h. The reaction was quenched with
5% aqueous citric acid solution and extracted with EtOAc, washed
subsequently with brine, dried over anhydrous MgSO.sub.4, filtered
and concentrated to produce 1-3 as an orange foam (2.31 g) that was
used without further purification. LCMS-ESI (m/z): [M].sup.+ calcd
for C.sub.35H.sub.49ClN.sub.4O.sub.7: 673.3. found: 673.7.
[0756] Step 4. Preparation of 1-4: To a solution of 1-3 (2.31 g,
3.43 mmol), TEA (0.72 mL, 5.15 mmol) and potassium
vinyltrifluoroborate (0.69 mg, 5.15 mmol) in EtOH (35 mL) was added
PdCl.sub.2(dppf) (0.25 g, 0.34 mmol, Frontier Scientific). The
reaction was sparged with Ar for 15 min and heated to 80.degree. C.
for 2 h. The reaction was adsorbed directly onto silica gel and
purified using silica gel chromatography to give 1-4 as a yellow
oil (1.95 g). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.53N.sub.4O.sub.7: 665.4. found: 665.3.
[0757] Step 5. Preparation of 1-5: To a solution of 1-4 (1.95 g,
2.93 mmol) in DCE (585 mL) was added Zhan 1B catalyst (0.215 g,
0.29 mmol, Strem) and the reaction was sparged with Ar for 15 min.
The reaction was heated to 80.degree. C. for 1.5 h, allowed to cool
to rt and concentrated. The crude product was purified by silica
gel chromatography to produce 1-5 as a yellow oil (1.47 g;
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.49N.sub.4O.sub.7: 637.4. found: 637.3).
[0758] Step 6. Preparation of 1-6: A solution of 1-5 (0.97 g, 1.52
mmol) in EtOH (15 mL) was treated with Pd/C (10 wt % Pd, 0.162 g).
The atmosphere was replaced with hydrogen and stirred at rt for 2
h. The reaction was filtered through Celite, the pad washed with
EtOAc and concentrated to give 1-6 as a brown foamy solid (0.803 g)
that was used subsequently without further purification.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.51N.sub.4O.sub.7: 639.4. found: 639.3.
[0759] Step 7. Preparation of 1-7: To a solution of 1-6 (0.803 g,
1.26 mmol) in DCM (10 mL) was added TFA (5 mL) and stirred at rt
for 3 h. An additional 2 mL TFA was added and the reaction stirred
for another 1.5 h. The reaction was concentrated to a brown oil
that was taken up in EtOAc (35 mL). The organic solution was washed
with water. After separation of the layers, sat. aqueous
NaHCO.sub.3 was added with stirring until the aqueous layer reached
a pH 7-8. The layers were separated again and the aqueous extracted
with EtOAc twice. The combined organics were washed with 1 M
aqueous citric acid, brine, dried over anhydrous MgSO.sub.4,
filtered and concentrated to produce 1-6 as a brown foamy solid
(0.719 g) that was used subsequently without further purification.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.31H.sub.43N.sub.4O.sub.7: 583.3. found: 583.4.
[0760] Step 8. Preparation of Example 1: To a solution of 1-7
(0.200 g, 0.343 mmol), Intermediate A10 (0.157 g, 0.515 mmol), DMAP
(0.063 g, 0.51 mmol) and DIPEA (0.3 mL, 1.72 mmol) in DMF (3 mL)
was added HATU (0.235 g, 0.617 mmol) and the reaction was stirred
at rt o/n. The reaction was diluted with MeCN and purified directly
by reverse phase HPLC (Gemini, 30-100% MeCN/H.sub.2O+0.1% TFA) and
lyophilized to give Example 1 (118.6 mg) as a solid TFA salt.
Analytic HPLC RetTime: 8.63 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.40H.sub.55F.sub.2N.sub.6O.sub.9S: 833.4. found:
833.5. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.19 (s, 1H); 7.80
(d, J=8.8 Hz, 1H); 7.23 (dd, J=8.8, 2.4 Hz, 1H); 7.15 (d, J=2.4 Hz,
1H); 5.89 (d, J=3.6 Hz, 1H); 5.83 (td, J.sub.H-F=55.6 Hz, J=6.4 Hz,
1H); 4.56 (d, J=7.2 Hz, 1H); 4.40 (s, 1H) 4.38 (ap d, J=7.2 Hz,
1H); 4.16 (dd, J=12, 4 Hz, 1H); 3.93 (s, 3H); 3.75 (dt, J=7.2, 4
Hz, 1H); 3.00-2.91 (m, 1H); 2.81 (td, J=12, 4.4 Hz, 1H); 2.63-2.54
(m, 1H); 2.01 (br s, 2H); 1.88-1.64 (m, 3H); 1.66-1.33 (m, 11H)
1.52 (s, 3H); 1.24 (t, J=7.2 Hz, 3H); 1.10 (s, 9H); 1.02-0.96 (m,
2H); 0.96-0.88 (m, 2H); 0.78-0.68 (m, 1H); 0.55-0.46 (m, 1H).
Example 2
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-(difluoromethyl)cyclopropyl]-9-ethyl-14-methoxy-3,6-dioxo-1,1a-
,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[-
18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamid-
e
##STR00214##
[0762] Example 2 was prepared in a similar fashion to Example 1,
substituting Intermediate A9 for Intermediate A10 in Step 8.
Example 2 was isolated (37.9 mg) in approximately 85% purity as a
TFA salt. Analytic HPLC RetTime: 8.54 min. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.39H.sub.53F.sub.2N.sub.6O.sub.9S:
819.35. found: 819.51. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
10.26 (s, 1H); 7.90 (d, J=9.2 Hz, 1H); 7.26 (dd, J=9.2, 2.4 Hz,
1H); 7.10 (d, J=2.4 Hz, 1H); 6.68 (br s, 1H); 6.01 (td,
J.sub.H-F=55.6 Hz, J=6.8 Hz, 1H); 5.87 (d, J=3.6 Hz, 1H); 5.38, (d,
J=10 Hz, 1H); 4.50-4.40 (m, 3H); 4.10 (dd, J=12, 3.6 Hz, 1H); 3.95
(s, 3H); 3.79-3.72 (m, 1H); 2.96-2.82 (m, 3H); 2.63-2.56 (m, 1H);
2.14 (t, J=6.8 Hz, 1H); 1.98-1.86 (m, 1H); 1.84-1.28 (m, 13H); 1.23
(t, J=7.2 Hz, 3H); 1.16-0.92 (m, 3H); 1.09 (s, 9H); 0.74-0.64 (m,
1H); 0.48 (q, J=6.4 Hz, 1H).
Example 3
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-{(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-ethylcyclopropyl}-9-ethyl-14-methoxy-3,6-dioxo-1,1a,3,4,5,6,9,-
10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1,10-
,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00215##
[0764] Example 3 was prepared in a similar fashion to Example 1,
substituting Intermediate A3 for Intermediate A10 in Step 8.
Example 3 was isolated (0.035 g) in approximately 88% purity as a
TFA salt. Analytic HPLC RetTime: 8.63 min. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.40H.sub.57N.sub.6O.sub.9S: 797.4.
found: 797.5. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.98 (s,
1H); 7.80 (d, J=9.2 Hz, 1H); 7.23 (d, J=9.2, 2.8 Hz, 1H); 7.15 (d,
J=2.8 Hz, 1H); 5.89 (d, J=3.6 Hz, 1H); 4.58 (d, J=7.6 Hz, 1H);
4.41-4.32 (m, 2H); 4.16 (dd, J=12.4 Hz, 3.6 Hz, 1H); 3.93 (s, 3H);
3.74 (dt, J=6.8, 2.8 Hz, 1H); 3.20-2.91 (m, 2H); 2.86-2.76 (m, 1H);
2.61-2.53 (m, 1H); 1.88-1.68 (m, 4H); 1.66-1.34 (m, 9H); 1.34-1.20
(m, 5H); 1.18-1.04 (m, 3H); 1.10 (s, 9H); 1.00-0.92 (m, 7H);
0.79-0.69 (m, 1H); 0.50 (br d, J=7.2 Hz, 1H).
Example 4
Preparation of (1
aR,5S,8S,9S,10R,22aR)-5-tert-butyl-9-ethyl-N-[(1R,2R)-2-ethyl-1-{[(1-meth-
ylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-3,6-dioxo-1,1a,3,-
4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,-
19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00216##
[0766] Example 4 was prepared in a similar fashion to Example 1,
substituting Intermediate A4 for Intermediate A10 in Step 8.
Example 4 was isolated (0.018 g) in approximately 88% purity as a
TFA salt. Analytic HPLC RetTime: 8.75. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.41H.sub.59N.sub.6O.sub.9S: 811.4. found: 811.6.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.91 (s, 1H); 7.80 (d,
J=9.2 Hz, 1H); 7.23 (dd, J=9.2, 2.8 Hz, 1H); 7.16 (d, J=2.8 Hz,
1H); 5.90 (d, J=3.6 Hz, 1H); 4.59 (d, J=6.8 Hz, 1H); 4.38 (s, 1H);
4.37 (d, J=11.6 Hz, 1H), 4.16 (dd, J=11.6, 6.8 Hz, 1H), 3.93 (s,
3H); 3.74 (dt, J=6.8, 3.6 Hz, 1H); 3.10-2.91 (m, 1H); 2.90-2.7 (m,
1H); 2.63-2.55 (m, 1H); 1.86-1.69 (m, 3H); 1.65-1.36 (m, 13H), 1.52
(s, 3H); 1.24 (t, J=7.2 Hz, 3H); 1.16-1.06 (m, 2H); 1.10 (s, 9H);
1.02-0.85 (m, 7H); 0.79-0.68 (m, 1H); 0.50 (br d, J=6.8 Hz,
1H).
Example 5
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-(difluoromethyl)cyclopropyl]-11-ethyl-16-methoxy-5,8-dioxo-1-
,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-1
OH-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-
-b]quinoxaline-10-carboxamide
##STR00217## ##STR00218## ##STR00219##
[0768] Step 1. Preparation of 5-1: HATU (555 mg, 1.46 mmol,
Oakwood) and DIPEA (1.10 mL, 6.35 mmol) were added to a mixture of
1-2 (533 mg, 1.20 mmol) and Intermediate D5 (414 mg, 1.33 mmol) in
12 mL of DMF under argon. After stirring overnight, the reaction
mixture was poured into water and extracted three times with ethyl
acetate. Combined organics were washed with water and brine, dried
(MgSO.sub.4), filtered, and concentrated under reduced pressure.
The resulting residue was purified by silica gel chromatography
(0-35% ethyl acetate in hexanes) to yield 5-1 (713 mg) as a white
solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.54ClN.sub.4O.sub.7: 701.36. found: 701.58.
[0769] Step 2. Preparation of 5-2:
Pd(dppf)Cl.sub.2*CH.sub.2Cl.sub.2 (94 mg, 0.115 mmol, Strem) was
added to a deoxygenated mixture of 5-1 (710 mg, 1.01 mmol),
potassium vinyltrifluoroborate (213 mg, 1.59 mmol), and
triethylamine (0.210 mL, 1.52 mmol) in 11 mL of EtOH at room
temperature. Reaction mixture was heated at 78.degree. C. under
argon for one hour. After cooling to room temperature, reaction
mixture was poured into water and extracted three times with ethyl
acetate. Combined organics were washed with water and brine, dried
(MgSO.sub.4), filtered, and concentrated under reduced pressure to
yield 5-2 (699 mg), which was used in the next step without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.57N.sub.4O.sub.7: 693.41. found: 693.47.
[0770] Step 3. Preparation of 5-3: A mixture of 5-2 (699 mg, 1.01
mmol) and Zhan 1B catalyst (81 mg, 0.111 mmol, Strem) in 200 mL of
DCE was deoxygenated under argon for 25 minutes. The mixture was
then heated at 95.degree. C. for 45 minutes. Reaction mixture was
heated at 95.degree. C. for 10 additional minutes, was cooled to
room temperature, and then concentrated under reduced pressure. The
resulting residue was purified by silica gel chromatography (0-30%
ethyl acetate in hexanes) to yield 5-3 (336 mg) as a light brown
solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.53N.sub.4O.sub.7: 665.38. found: 665.53.
[0771] Step 4. Preparation of 5-4: Palladium on carbon (10 wt. %
Pd, 102 mg, 0.096 mmol) was added to a solution of 5-3 (330 mg,
0.497 mmol) in 8 mL of ethanol and 3.5 mL of ethyl acetate. Mixture
was stirred under an atmosphere of hydrogen for 100 minutes and was
then filtered over Celite, washing with ethyl acetate. Filtrate was
concentrated under reduced pressure to yield 5-4 (64 mg) as a light
yellow-brown solid film, which was used in the next step without
further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.55N.sub.4O.sub.7: 667.40. found: 667.52.
[0772] Step 5. Preparation of 5-5: TMSOTf (0.53 mL, 2.91 mmol) was
added dropwise to a solution of 5-4 (329 mg, 0.494 mmol) in 10 mL
of dichloromethane under argon at room temperature. After one hour,
an additional 0.3 mL of TMSOTf was added. After an additional hour,
reaction mixture was concentrated under reduced pressure. The
resulting film was taken up in 12 mL of toluene and concentrated
under reduced pressure. This process was repeated a second time to
yield 5-5 (301 mg), which was used in the next step without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.33H.sub.47N.sub.4O.sub.7: 611.34. found: 611.46.
[0773] Step 6. Preparation of Example 5: HATU (129 mg, 0.339 mmol)
and DIPEA (0.22 mL, 1.27 mmol) were added to a mixture of 5-5 (134
mg, 0.22 mmol) and Intermediate A9 (95 mg, 0.328 mmol) in 6.6 mL of
MeCN under argon. After stirring for 5 h, reaction mixture was
poured into water and extracted three times with ethyl acetate.
Combined organics were washed with water and brine, dried
(MgSO.sub.4), filtered, and concentrated under reduced pressure.
The resulting residue was purified by reverse phase preparatory
HPLC (15-100% acetonitrile in water, with 0.1% trifluoroacetic acid
buffer) to yield Example 5 (43 mg) as a light yellow solid,
trifluoroacetic acid salt, after lyophilization. Analytic HPLC
RetTime: 9.11 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S: 847.38. found: 847.62.
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 9.31 (s, 1H), 7.80 (d,
J=9.2 Hz, 1H), 7.23 (dd, J=15.4, 2.8 Hz, 1H), 7.19 (d, J=2.8 Hz,
1H), 5.87 (td, J.sub.H-F=56 Hz, J=6 Hz, 1H), 5.87-5.83 (m, 1H),
4.59 (d, J=7.6 Hz, 1H), 4.38 (s, 1H), 4.23-4.14 (m, 2H), 3.93 (s,
3H), 3.06-2.94 (m, 2H), 2.77-2.67 (m, 1H), 2.65-2.58 (m, 1H),
2.07-2.01 (m, 2H), 1.98-1.74 (m, 4H), 1.72-1.52 (m, 4H), 1.50-1.20
(m, 12H), 1.18-1.02 (m, 8H), 1.06 (s, 9H).
Example 6
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[-
(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-1-ethyl-16-methoxy-5,-
8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-10H-9,12-m-
ethanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxa-
line-10-carboxamide
##STR00220##
[0775] Example 6 was prepared in a similar fashion to Example 5,
substituting Intermediate A10 for Intermediate A9 in Step 6.
Example 6 was isolated (29 mg) as a white solid. Analytic HPLC
RetTime: 9.26 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.59F.sub.2N.sub.6O.sub.9S: 861.40. found: 861.20.
.sup.1H NMR (400 MHz, CDCl.sub.3) b 9.91 (s, 1H), 7.82 (d, J=12 Hz,
1H), 7.18 (d, J=12 Hz 1H), 7.13-7.06 (m, 1H), 6.48 (s, 1H), 5.95
(td, J.sub.H-F=56 Hz, J=6 Hz, 1H), 5.82 (d, J=4.4 Hz, 1H), 5.33 (d,
J=10 Hz, 1H), 4.95-4.91 (m, 1H), 4.38-4.31 (m, 2H), 4.10-3.88 (m,
2H), 3.98 (s, 3H), 2.98-2.89 (m, 1H), 2.67-2.59 (m, 1H), 2.05-1.65
(m, 4H), 1.64-1.21 (m, 12H), 1.40 (s, 3H), 1.17-0.80 (m, 12H), 1.09
(s, 9H).
Example 7
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-1a-m-
ethyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,1-
0-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quin-
oxaline-8-carboxamide
##STR00221## ##STR00222##
[0777] Step 1. Preparation of 1-2 (free base): Carbamate 1-1 (350
mg, 0.689 mmol) was added to a flask containing a 4:1 mixture of
t-butyl acetate:DCM (3.5 mL). To this solution was then added
methanesulfonic acid (447 .mu.L, 6.89 mmol). The reaction mixture
was allowed to stir for 20 min at rt, then diluted with methylene
chloride (20 mL) and saturated aqueous sodium bicarbonate (20 mL).
The solution was allowed to stir until evolution of gas ceased,
then the organics were removed and the aqueous layer was extracted
twice with methylene chloride (20 mL). The combined organics were
then washed with brine, dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The resulting white solid 1-2 (free base,
280 mg) was used in the subsequent reaction without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.20H.sub.27ClN.sub.3O.sub.4: 408.2. found: 408.1.
[0778] Step 2. Preparation of mixture 7-1: Amine 1-2 (281 mg, 0.689
mmol) was combined with diastereomeric Intermediate mixture D6 (266
mg, 0.895 mmol), DIPEA (600 .mu.L, 3.45 mmol) and DMF (2 mL). HATU
(340 mg, 0.895 mmol) was then added to the reaction mixture, which
was stirred at 40.degree. C. for 5 h. Reaction mixture was then
diluted with water (10 mL) and taken up into methylene chloride (10
mL). Organics were separated and aqueous layer was extracted once
with methylene chloride (10 mL). Combined organics were then washed
with brine, dried over MgSO.sub.4, filtered, and concentrated in
vacuo. Crude residue was then purified via silica gel
chromatography to give 7-1 as a 1:1 diastereomeric mixture (280
mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.52ClN.sub.4O.sub.7: 687.4. found: 687.3.
[0779] Step 3. Preparation of 7-2: Pd(dppf)Cl.sub.2 (29 mg, 0.0407
mmol) was added to a degassed mixture of 7-1 (280 mg, 0.407 mmol),
potassium vinyltrifluoroborate (55 mg, 0.733 mmol), and
triethylamine (91 .mu.L, 0.651 mmol) in 2 mL of ethanol at room
temperature. Reaction mixture was heated at 80.degree. C. under
N.sub.2 for one hour. After cooling to room temperature, reaction
mixture was diluted with toluene (10 mL), concentrated in vacuo to
a small volume of solvent, and rediluted in toluene (1 mL). Mixture
was then loaded directly onto a silica column and purified by
silica gel chromatography to afford 7-2 as a 1:1 diastereomeric
mixture which was carried on to the next step without concentrating
fully to dryness. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.55N.sub.4O.sub.7: 679.4. found: 679.4.
[0780] Step 4. Preparation of 7-3 and 7-4: Diastereomeric mixture
7-2 (276 mg, 0.407 mmol) and Zhan 1B catalyst (32 mg, 0.0407 mmol,
Strem) were dissolved in 80 mL of DCE and degassed under N.sub.2
for 25 minutes. The mixture was then heated to 100.degree. C. for 1
h. Reaction was then cooled to room temperature and concentrated in
vacuo. The resulting residue was purified via silica gel
chromatography (0% to 30% ethyl acetate in hexanes) to yield single
diastereomers 7-3 (20 mg, early eluting fraction) and 7-4 (25 mg,
late eluting fraction) as light brown residues. Early eluting
fraction: LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.51N.sub.4O.sub.7: 651.4. found: 651.3. Late eluting
fraction: LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.51N.sub.4O.sub.7: 651.4. found: 651.3.
[0781] Step 5. Preparation of 7-5: Palladium on carbon (10% w/w, 25
mg) was added to a solution of 7-3 (20 mg, 0.0307 mmol) in a 1:1
mixture of ethyl acetate and dioxane (2 mL). Mixture was stirred
under an atmosphere of hydrogen for 30 min and was then filtered
through a plug of Celite, and washed with ethyl acetate. Filtrate
was concentrated under reduced pressure to yield 7-5 (16 mg) as a
light brown film, which was used in the next step without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.53N.sub.4O.sub.7: 653.4. found: 653.4.
[0782] Step 6. Preparation of 7-6: Intermediate 7-5 (16 mg, 0.023
mmol) was dissolved in 2 M HCl in dioxane (2 mL) and heated at
80.degree. C. for 1.5 h via microwave reactor. Reaction mixture was
then concentrated in vacuo to give 7-6 (15 mg) as a brown residue,
which was used in the subsequent step without further purification.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.44N.sub.4O.sub.7: 597.3. found: 597.3.
[0783] Step 7. Preparation of Example 7: HATU (11.9 mg, 0.031 mmol)
and DIPEA (22 .mu.L, 0.126 mmol) were added to a mixture of 7-6 (15
mg, 0.025 mmol) and A10 (11.5 mg, 0.0377 mmol) in 1 mL of DMF.
After stirring overnight at room temperature, reaction mixture was
poured into water, acidified to pH 1 with 1 N aqueous HCl, and
extracted three times with methylene chloride (15 mL). Combined
organics were washed with water, brine, dried over MgSO.sub.4,
filtered, and concentrated under reduced pressure. The resulting
residue was purified by reverse phase prep HPLC (5-100%
acetonitrile in water, with 0.1% trifluoroacetic acid buffer)
followed by silica gel chromatography to afford Example 7 (4.3 mg)
as a white solid film. Analytic HPLC RetTime: 9.07 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S: 847.4. found: 847.4.
.sup.1H NMR (400 MHz, CDCl.sub.3) (9.88 (s, 1H), 7.83 (d, J=9.1 Hz,
1H), 7.20 (dd, J=9.1 Hz, 2.8 Hz, 1H), 7.07 (d, J=2.7 Hz, 1H), 6.56
(s, 1H), 5.98 (td, J.sub.H-F=55.7, J=6.7 Hz, 1H), 5.95 (d, J=9.6,
1H), 5.32 (d, J=9.6 Hz, 1H), 4.45 (dd, J=13.0 Hz, 9.6 Hz, 2H), 4.32
(d, J=9.7 Hz, 1H), 4.13 (dd, J=15.5 Hz, 8.8 Hz, 1H), 3.93 (s, 3H),
2.99-2.84 (m, 1H), 2.82-2.68 (m, 1H), 2.62-2.47 (m 1H), 2.16-2.02
(m, 1H) 2.00-1.85 (m, 1H) 1.84-1.69 (m, 1H), 1.70-1.15 (m, 11H),
1.52 (s, 3H), 1.50 (s, 3H), 1.20 (t, J=7.3 Hz, 3H), 1.14-0.77 (m,
5H) 1.09 (s, 9H), 0.11 (m, 1H).
Example 8
Preparation of
(1aS,5S,8S,9S,10R,22aS)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-1a-m-
ethyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,1-
0-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quin-
oxaline-8-carboxamide
##STR00223##
[0785] Example 8 was prepared in a similar fashion to Example 7,
substituting late eluting 7-4 for early eluting 7-3 in Step 5.
Example 7 was isolated (2.9 mg) as a white solid. Analytic HPLC
RetTime: 9.09 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S: 847.4. found: 847.4.
Examples 9 and 10
Preparation of
(7S,10S,11S,12R)-7-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-methyl-
cyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-11-ethyl-16-methoxy-5,8-dioxo--
3aR-(trifluoromethyl)-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahy-
dro-10H-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[1-
1,12-b]quinoxaline-10-carboxamide and
(7S,10S,11S,12R)-7-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-methyl-
cyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-11-ethyl-16-methoxy-5,8-dioxo--
3aS-(trifluoromethyl)-1,2,3,3a,5,6,7,8,
11,12,20,21,22,23,24,24a-hexadecahydro-10H-9,12-methanocyclopenta[18,19][-
1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-10-carboxamide
##STR00224## ##STR00225## ##STR00226##
[0787] Step 1. Preparation of 9-1: To a solution of Intermediate D8
(322 mg, 0.85 mmol) and 1-2 (316 mg, 0.78 mmol) in MeCN (3.9 mL)
was added HATU (323 mg, 0.85 mmol) followed by DIPEA (678 .mu.L,
3.90 mmol) at rt under an argon atmosphere. After 2 h, the reaction
mixture was concentrated in vacuo, and the crude residue was
purified by silica gel chromatography (0-100% ethyl acetate/hexanes
gradient) to afford amide 9-1 (476 mg, 1:1 diastereomeric mixture)
as a colorless oil. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.53ClF.sub.3N.sub.4O.sub.7: 769.4. found: 769.5.
[0788] Step 2. Preparation of 9-2: To a solution of 9-1 (470 mg,
612 .mu.mol), TEA (128 .mu.L, 918 .mu.mol), and potassium
vinyltrifluoroborate (123 mg, 918 .mu.mol) in EtOH (3.06 mL) was
added PdCl2(dppf) (50 mg, 61 .mu.mol). The reaction mixture was
deoxygenated with argon for 10 min and heated to 78.degree. C.
After 1 h, the reaction mixture was allowed to cool to rt and was
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes gradient) to afford
vinyl quinoxaline 9-2 (329 mg, 1:1 diastereomeric mixture) as a
yellow oil. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.56F.sub.3N.sub.4O.sub.7: 761.4. found: 761.6.
[0789] Step 3. Preparation of 9-3: To a solution of 9-2 (329 mg,
485 .mu.mol) in DCE (97 mL) was added Zhan 1B catalyst (35 mg, 49
.mu.mol, Strem) and the reaction mixture was deoxygenated for 10
minutes with argon. The reaction mixture was then heated to
100.degree. C. After 30 min, the reaction mixture was allowed to
cool to rt and was concentrated in vacuo. The crude residue was
purified by silica gel chromatography (0-100% ethyl acetate/hexanes
gradient) to afford macrocycle 9-3 (301 mg, 7:4 diastereomeric
mixtures) as a light yellow oil. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.38H.sub.52F.sub.3N.sub.4O.sub.7: 733.4. found:
733.5.
[0790] Step 4. Preparation of 9-4: To a solution of 9-3 (300 mg,
410 .mu.mol) in ethanol (2.00 mL) was added Pd/C (10 wt % Pd, 43
mg, 41 .mu.mol) at rt under an argon atmosphere. The atmosphere of
the reaction was replaced with hydrogen gas and the reaction
mixture stirred vigorously at rt. After 30 min, the reaction
mixture was diluted with ethyl acetate (10 mL) and filtered through
a pad of Celite with ethyl acetate washings (3.times.5 mL). The
filtrate was concentrated in vacuo to afford macrocycle 9-4 (295
mg, 7:4 diastereomeric mixture), which was used directly in the
next step without further purification. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.38H.sub.54F.sub.3N.sub.4O.sub.7: 735.4. found:
735.5.
[0791] Step 5. Preparation of 9-5: To a solution of 9-4 (295 mg,
401 .mu.mol) in DCM (2 mL) was added TMSOTf (72.6 .mu.L, 401 mmol)
at rt under an argon atmosphere. After 1.5 h, additional TMSOTf
(362.9 .mu.L, 2.00 mmol) was added. After 1 h, additional TMSOTf
(362.9 .mu.L, 2.00 mmol) was added. After 2 h, the reaction mixture
was added slowly to a 0.25 N aqueous NaOH solution (precooled to
0.degree. C., 3 mL). The resulting mixture was diluted with 1 N
aqueous HCl solution (5 mL), and was extracted with DCM (3.times.5
mL). The combined organic extracts were dried over anhydrous sodium
sulfate and were concentrated to afford carboxylic acid 9-5 (353
mg, 7:4 diastereomeric mixture) as a tan solid, which was used
directly in the next step without further purification.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.45F.sub.3N.sub.4O.sub.7: 679.3. found: 679.5.
[0792] Step 6. Preparation of Example 9 and Example 10: To a
solution of acid 9-5 (150 mg, 220 .mu.mol) and Intermediate A10
(101 mg, 330 .mu.mol) in MeCN (1.1 mL) was added HATU (127 mg, 330
.mu.mol) followed by DIPEA (191 .mu.L, 1.10 mmol) at rt under an
argon atmosphere. After 1 h, the reaction mixture was concentrated
in vacuo, and the crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes gradient). The
fractions containing the desired product were combined and were
repurified by silica gel chromatography (0-50% acetone/hexanes
gradient) to afford the first eluting
[0793] Example 9 (40 mg) as a white powder and the second eluting
Example 10 (70 mg) as a white powder. First eluting Example 9:
Analytic HPLC RetTime: 9.42 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.43H.sub.58F.sub.5N.sub.6O.sub.9S: 929.4. found: 929.5.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.83 (s, 1H), 7.92 (d,
J=9.1 Hz, 1H), 7.19 (dd, J=9.0, 2.6 Hz, 1H), 7.13 (d, J=2.6 Hz,
1H), 5.99 (br s, 1H), 5.96 (td, J.sub.H-F 55.5, J=6.6 Hz, 1H), 5.70
(d, J=10.0 Hz, 1H), 4.63 (d, J=6.6 Hz, 1H), 4.38 (d, J=10.0 Hz,
1H), 4.22-4.04 (m, 2H), 3.96 (s, 3H), 3.12-2.89 (m, 1H), 2.71-2.51
(m, 2H), 2.17 (s, 3H), 2.15-1.82 (m, 4H), 1.83-1.34 (m, 8H),
1.36-0.98 (m, 12H), 1.26 (s, 9H), 0.92-0.79 (m, 4H). Second eluting
Example 10: Analytic HPLC RetTime: 9.55 min. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.43H.sub.58F.sub.5N.sub.6O.sub.9S:
929.4. found: 929.5. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.61
(s, 1H), 7.91 (d, J=9.1 Hz, 1H), 7.23 (dd, J=9.0, 3.0 Hz, 1H), 7.18
(d, J=2.7 Hz, 1H), 5.98-5.91 (m, 1H), 5.83 (td, J.sub.H-F 55.5,
J=6.6 Hz, 1H), 5.33 (d, J=9.8 Hz, 1H), 4.72-4.63 (m, 1H), 4.46-4.38
(m, 1H), 4.32 (d, J=10.0 Hz, 1H), 4.25-4.14 (m, 1H), 3.97 (s, 3H),
3.73 (br d, J=7.6 Hz, 1H), 3.23-3.07 (m, 1H), 2.86-2.37 (m, 2H),
2.14-1.79 (m, 2H), 1.78-1.38 (m, 8H), 1.51 (s, 3H), 1.35-1.08 (m,
8H), 1.25 (s, 9H), 1.05 (br s, 3H), 0.93-0.68 (m, 6H).
Examples 11 and 12
Preparation of
(7S,10S,11S,12R)-7-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl)carbamoy-
l]-2-(difluoromethyl)cyclopropyl]-11-ethyl-16-methoxy-5,8-dioxo-3aR-(trifl-
uoromethyl)-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-1
OH-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-
-b]quinoxaline-10-carboxamide and
(7S,10S,11S,12R)-7-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl)carbamoy-
l]-2-(difluoromethyl)cyclopropyl]-11-ethyl-16-methoxy-5,8-dioxo-3aS-(trifl-
uoromethyl)-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-1
OH-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-
-b]quinoxaline-10-carboxamide
##STR00227##
[0795] Preparation of Example 11 and Example 12: To a solution of
acid 9-5 (150 mg, 220 .mu.mol) and Intermediate A9 (96 mg, 330
.mu.mol) in MeCN (1.1 mL) was added HATU (127 mg, 330 .mu.mol)
followed by DIPEA (191 .mu.L, 1.10 mmol) at rt under an argon
atmosphere. After 1 h, the reaction mixture was concentrated in
vacuo, and the crude residue was purified by silica gel
chromatography (0-50% acetone/hexanes gradient). The fractions
containing the desired product were combined and were repurified by
silica gel chromatography (0-50% acetone/hexanes gradient) to
afford the first eluting Example 11 (29 mg) as a white powder and
the second eluting Example 12 (60.2 mg) as a white powder. First
eluting Example 11: Analytic HPLC RetTime: 9.44 min. LCMS-ESI
(m/z): [M+H].sup.+ calcd C.sub.42H.sub.56F.sub.5N.sub.6O.sub.9S:
915.4. found: 915.6. .sup.1H NMR (400 MHz, CDCl.sub.3) b 10.17 (br
s, 1H), 7.83 (d, J=9.1 Hz, 1H), 7.21 (dd, J=9.1, 2.7 Hz, 1H),
7.17-7.07 (m, 1H), 5.99 (br s, 1H), 5.97 (td, J.sub.H-F 55.5, J=6.6
Hz, 1H), 5.82 (d, J=9.8 Hz, 1H), 4.55 (d, J=7.2 Hz, 1H), 4.39 (d,
J=10.0 Hz, 1H), 4.20-4.03 (m, 2H), 3.95 (s, J=5.9 Hz, 3H),
2.97-2.82 (m, 2H), 2.79-2.49 (m, 3H), 2.24-1.81 (m, 8H), 1.80-1.11
(m, 12H), 1.10-0.98 (m, 4H), 1.07 (s, 9H), 0.95-0.81 (m, 3H).
Second eluting Example 12: Analytic HPLC RetTime: 9.48 min.
LCMS-ESI (m/z): [M+H].sup.+ calcd
C.sub.42H.sub.56F.sub.5N.sub.6O.sub.9S: 915.4. found: 915.6.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.07 (s, 1H), 7.93 (d,
J=9.6 Hz, 1H), 7.28-7.20 (m, 1H), 7.16 (s, 1H), 6.17-5.68 (m, 3H),
4.67-4.55 (m, 1H), 4.37-4.23 (m, 2H), 4.17-4.05 (m, 1H), 3.97 (s,
3H), 3.75-3.66 (m, 1H), 3.22-3.04 (m, 1H), 3.02-2.31 (m, 6H),
2.30-1.83 (m, 10H), 1.85-1.13 (m, 13H), 1.06 (s, 9H), 0.95-0.79 (m,
1H).
Example 13
Preparation of
(1R,4S,4aR,8S,11S,12S,13R,25aR)-8-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsu-
lfonyl)carbamoyl]-2-(difluoromethyl)cyclopropyl]-12-ethyl-17-methoxy-6,9-d-
ioxo-2,3,4,4a,6,7,8,9,12,13,21,22,23,24,25,25a-hexadecahydro-1H,11H-1,4:10-
,13-dimethanoquinoxalino[2,3-k][1,10,3,6]benzodioxadiazacyclononadecine-11-
-carboxamide
##STR00228## ##STR00229##
[0797] Step 1. Preparation of diastereomer mixture 13-1 and 13-2:
To a solution of 1-2 (354 mg, 0.87 mmol), Intermediate mixture D9
and D10 (323 mg, 0.96 mmol) and BEP (263 mg, 0.96 mmol; TCI
America) was added DIPEA (0.45 mL, 2.61 mmol) and the reaction was
stirred at 50.degree. C. for 2 h. The reaction was quenched with
sat. aqueous NaHCO.sub.3 solution and extracted with EtOAc, the
organic phase was washed with brine, dried over magnesium sulfate
and concentrated. The crude product was purified by silica gel
chromatography (0-30% EtOAc/hexanes) to yield an inseparable
mixture of diastereomers 13-1 and 13-2 (338 mg). LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.39H.sub.56ClN.sub.4O.sub.7: 727.38.
found: 727.46.
[0798] Step 2. Preparation of diastereomer mixture 13-3 and 13-4:
To a solution of the mixture of 13-1 and 13-2 (338 mg, 0.46 mmol),
TEA (0.10 mL, 0.69 mmol) and potassium vinyltrifluoroborate (93 mg,
0.69 mmol) in EtOH (30 mL) was added PdCl.sub.2(dppf) (38 mg, 0.046
mmol, Strem Chemicals). The reaction was deoxygenated with N.sub.2
for 10 min and heated to 80.degree. C. for 1 h. The reaction was
quenched with sat. aqueous NaHCO.sub.3 solution and extracted with
EtOAc, washed subsequently with brine, dried over magnesium sulfate
and concentrated. The residue was purified using silica gel
chromatography to give an inseparable mixture of diastereomers 13-3
and 13-4 (285 mg). LCMS-ESI+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.59N.sub.4O.sub.7: 719.44. found: 719.70.
[0799] Step 3 and 4. Preparation of 13-5: To a solution of the
diastereomeric mixture 13-3 and 13-4 (285 mg, 0.40 mmol) in DCE
(100 mL) was added Zhan 1B catalyst (30 mg, 0.04 mmol, Strem) and
the reaction was deoxygenated for 30 minutes with N.sub.2. The
reaction was heated to 100.degree. C. for 45 min, allowed to cool
to rt and concentrated. The crude product was purified by silica
gel chromatography to produce macrocyclic olefin product (125 mg;
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.55N.sub.4O.sub.7: 691.41. found: 691.58) that was
taken up in EtOH (6 mL) and treated with Pd/C (10%, 120 mg). The
atmosphere was replaced with hydrogen and stirred at rt for 1.5 h.
The reaction was filtered over Celite, washed with EtOAc and
concentrated to give 13-5 as an oil (125 mg) that was used
subsequently without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.39H.sub.57N.sub.4O.sub.7: 693.42.
found: 693.46.
[0800] Step 5. Preparation of 13-6: To a solution of 13-5 (50 mg,
0.072 mmol) in DCM (4 mL) was added TFA (1 mL) and stirred at rt
for 6 h. The reaction was diluted with EtOAc, washed with H.sub.2O,
aqueous pH 7 buffer, dried over magnesium sulfate, and concentrated
to give 13-6 as a residue that was used subsequently without
further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.49N.sub.4O.sub.7: 637.36. found: 637.40.
[0801] Step 6. Preparation of Example 13: To a solution of 13-6 (46
mg, 0.072 mmol), Intermediate A9 (28 mg, 0.11 mmol), TBTU (34 mg,
0.10 mmol) and DMAP (13 mg, 0.11 mmol) in DCM (5 mL) was added
DIPEA (0.038 mL, 0.22 mmol) and the reaction was stirred at rt for
16 h. The reaction was quenched with water, diluted with EtOAc,
washed with sat. aqueous NaHCO.sub.3, brine, dried over magnesium
sulfate, and concentrated. The crude material was purified by
reverse phase HPLC (Gemini, 30-85% MeCN/H.sub.2O+0.1% TFA) and
lyophilized to give Example 13 (14.5 mg) as a TFA salt. Analytic
HPLC RetTime: 9.39 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.43H.sub.59F.sub.2N.sub.6O.sub.9S: 873.40. found: 873.42.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.28 (s, 1H), 7.82 (d,
J=9.2 Hz, 1H), 7.26 (dd, J=6.4, 2.8 Hz, 1H), 7.19 (d, J=2.8 Hz,
1H), 6.04-5.74 (m, 2H), 5.50 (s, 1H), 4.55 (d, J=7.6 Hz, 1H), 4.47
(s, 1H), 4.26-4.16 (m, 2H), 3.94 (s, 3H), 3.03-2.95 (m, 2H),
2.78-2.66 (m, 2H), 2.17 (br, 2H), 2.05 (s, 3H), 1.90-1.85 (m, 1H),
1.76-1.74 (m, 2H), 1.61-1.21 (m, 20H), 1.15-1.11 (m, 2H), 1.08 (s,
9H), 0.93-0.90 (m, 1H).
Example 14
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-cyclopentyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(-
1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-3,6-
-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methano-
cyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-
-carboxamide
##STR00230## ##STR00231## ##STR00232##
[0803] Step 1. Preparation of 14-1: To a solution of 1-2 (223 mg,
0.50 mmol) and Intermediate D2 (221 mg, 0.75 mmol) in acetonitrile
(5 mL) was added HATU (306 mg, 0.80 mmol) followed by DIPEA (0.43
mL, 2.5 mmol) at room temperature. After 19 h, solvent was removed
under reduced pressure and the resulting residue was diluted with
ethyl acetate (15 mL). The resulting solution was washed with 1 M
aqueous HCl (10 mL). The aqueous layer was extracted with ethyl
acetate (2.times.10 mL) and combined organic layer was washed with
brine (15 mL), dried over anhydrous magnesium sulfate and
concentrated. The resulting crude residue was purified via silica
gel chromatography (0-100% ethyl acetate/hexanes gradient) to
afford 14-1 (173 mg) as colorless oil. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.36H.sub.50ClN.sub.4O.sub.7: 685.33. found:
685.49.
[0804] Step 2. Preparation of 14-2: To a solution of 14-1 (173 mg,
0.25 mmol) in EtOH (3 mL) was added potassium vinyltrifluoroborate
(51 mg, 0.38 mmol), PdCl.sub.2(dppf) (21 mg, 0.025 mmol) and TEA
(0.053 mL, 0.38 mmol) sequentially and the resulting mixture was
heated to 80.degree. C. After 1 h, additional potassium
vinyltrifluoroborate (17 mg, 0.12 mmol) was added and continued
stirring at 80.degree. C. After 2.5 h, additional potassium
vinyltrifluoroborate (8 mg, 0.06 mmol) was added and the reaction
was stirred for additional 10 minutes at 80.degree. C. The reaction
was cooled to room temperature, diluted with ethyl acetate (20 mL),
and washed with brine (20 mL). Aqueous layer was extracted with
ethyl acetate (10 mL), and the combined organic layer was dried
over anhydrous magnesium sulfate and concentrated to afford 14-2 as
a residue which was used it without purification in the next step.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.53N.sub.4O.sub.7: 677.38. found: 677.50.
[0805] Step 3. Preparation of 14-3: To a solution of 14-2 in
deoxygenated DCE (0.006 M) was added Zhan 1B catalyst (18 mg, 0.025
mmol, Strem) and the reaction was deoxygenated for another 10
minutes with Ar. The reaction was heated to 100.degree. C. After
1.5 h, Zhan 1B catalyst (9 mg, 0.012 mmol) was added and the
reaction was stirred for another 30 min. The reaction mixture was
allowed to cool to rt and concentrated to 4-5 mL volume. This was
directly purified by silica gel chromatography to afford 14-3 as a
brown oil (70 mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49N.sub.4O.sub.7: 649.35. found: 649.50.
[0806] Step 4. Preparation of 14-4: To a solution of 14-3 (70 mg,
0.11 mmol) in EtOH (5 mL) was added Pd/C (10 wt % Pd, 12 mg) under
argon. The atmosphere was replaced with hydrogen and the reaction
was stirred at rt for 16 h. The reaction was filtered over Celite,
washed with EtOH and concentrated to give 14-4 as a brown oil that
was used subsequently without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.36H.sub.51N.sub.4O.sub.7: 651.37.
found: 651.60.
[0807] Step 5. Preparation of 14-5: To a solution of 14-4 (70 mg,
0.11 mmol) in DCM (3 mL) was added TMSOTf (0.103 mL, 0.53 mmol) and
the reaction was stirred at rt for 1 h. The reaction was
concentrated to afford 14-5 which was used it for the next step
without purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.43N.sub.4O.sub.7: 595.31. found: 595.43.
[0808] Step 6. Preparation of Example 14: To a solution of 14-5
(36.8 mg, 0.06 mmol) and Intermediate A10 (28 mg, 0.09 mmol) in
acetonitrile (1.5 mL) was added HATU (38 mg, 0.1 mmol) followed by
DIPEA (0.065 mL, 0.37 mmol) at room temperature. After 20 minutes,
the reaction mixture was directly purified by reverse phase HPLC
(Gemini 5u C18 110 .ANG. column, 15-100% MeCN/H.sub.2O+0.1% TFA)
and lyophilized to afford Example 14 as a yellow solid (24 mg) as a
TFA salt. Analytic HPLC RetTime: 9.03 min. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.41H.sub.55F.sub.2N.sub.6O.sub.9S:
845.4. found: 845.6. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.31
(s, 1H), 7.80 (d, J=9.1 Hz, 1H), 7.23 (dd, J=9.1, 2.8 Hz, 1H), 7.16
(d, J=2.7 Hz, 1H), 6.03-5.66 (m, 2H), 4.53 (dd, J=13.2, 9.6 Hz,
2H), 4.18 (dd, J=17.2, 7.1 Hz, 2H), 3.92 (s, 3H), 3.68 (dt, J=6.8,
2.8 Hz, 1H), 3.13 (quin, J=1.7 Hz, 1H), 3.02-2.92 (m, 1H),
2.85-2.78 (m, 1H), 2.62-2.55 (m, 1H), 2.30-2.17 (m, 1H), 2.02 (s,
2H), 1.97-1.86 (m, 3H), 1.86-1.79 (m, 1H), 1.80-1.41 (m, 17H),
1.40-1.28 (m, 3H), 1.22 (t, J=7.4 Hz, 3H), 1.03-0.87 (m, 4H),
0.76-0.68 (m, 1H), 0.51-0.44 (m, 1H).
Example 15
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-cyclopentyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl)-
carbamoyl]-2-(difluoromethyl)cyclopropyl]-9-ethyl-14-methoxy-3,6-dioxo-1,1-
a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa-
[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxami-
de
##STR00233##
[0810] Step 1. Preparation of Example 15. To a solution of 14-5 (27
mg, 0.045 mmol) and Intermediate A9 (20 mg, 0.067 mmol) in
acetonitrile (1.3 mL) was added HATU (27 mg, 0.072 mmol) followed
by DIPEA (0.047 mL, 0.27 mmol) at room temperature. After 20
minutes, the reaction mixture was directly purified by reverse
phase HPLC (Gemini 5u C18 110 .ANG. column, 15-100%
MeCN/H.sub.2O+0.1% TFA) and lyophilized to afford Example 15 as a
yellow solid (18.6 mg) as a TFA salt. Analytic HPLC RetTime: 8.89
min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.53F.sub.2N.sub.6O.sub.9S: 831.4. found: 831.6.
.sup.1H NMR (400 MHz, CD.sub.3OD) (9.32 (s, 1H), 7.79 (d, J=9.1 Hz,
1H), 7.23 (dd, J=9.1, 2.8 Hz, 1H), 7.16 (d, J=2.8 Hz, 1H),
6.03-5.66 (m, 2H), 4.53 (t, J=10.0 Hz, 2H), 4.22-4.14 (m, 2H), 3.92
(s, 3H), 3.67 (dt, J=6.5, 2.9 Hz, 1H), 3.13 (quin, 1.6 Hz, 1H),
3.04-2.92 (m, 3H), 2.85-2.77 (m, 1H), 2.63-2.55 (m, 1H), 2.26-2.19
(m, 1H), 2.05-2.02 (m, 2H), 1.99-1.86 (m, 3H), 1.84-1.42 (m, 12H),
1.41-1.25 (m, 4H), 1.22 (t, J=7.2 Hz, 3H), 1.15-1.03 (m, 3H),
1.01-0.90 (m, 2H), 0.76-0.68 (m, 1H), 0.49-0.45 (m, 1H).
Example 16
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-cyclohexyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-3,6--
dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanoc-
yclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8--
carboxamide
##STR00234## ##STR00235## ##STR00236##
[0812] Step 1. Preparation of 16-1: To a solution of Intermediate
D3 (190 mg, 0.60 mmol) and 1-2 (264 mg, 0.60 mmol) in DMF (5 mL)
was added DIPEA (0.31 mL, 1.8 mmol) followed by COMU (257 mg, 0.60
mmol) at rt. After 2 h, the solvent was removed under reduced
pressure and the resulting residue diluted with ethyl acetate (15
mL). The resulting solution was washed with 10% aqueous citric acid
solution. The aqueous layer was extracted with ethyl acetate
(2.times.10 mL) and combined organic layer was washed with brine
(15 mL), dried over anhydrous magnesium sulfate and concentrated.
The resulting crude residue was purified via silica gel
chromatography to afford 16-1 (260 mg) as a colorless oil. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.37H.sub.51ClN.sub.4O.sub.7:
700.28. found: 700.03.
[0813] Step 2. Preparation of 16-2: To a solution of 16-1 (260 mg,
0.37 mmol) in EtOH (5 mL) were added potassium vinyltrifluoroborate
(75 mg, 0.56 mmol), PdCl.sub.2(dppf) (30 mg, 0.037 mmol) and TEA
(0.079 mL, 0.56 mmol) sequentially. The reaction was deoxygenated
with Ar for 12 min and was heated to 78.degree. C. for 2 h. The
reaction was cooled to rt, diluted with ethyl acetate (20 mL), and
washed with brine (20 mL). The aqueous layer was extracted with
ethyl acetate (10 mL), and the combined organic layers were dried
over anhydrous magnesium sulfate and concentrated to afford crude
residue. The resulting crude residue was purified via silica gel
chromatography to afford 16-2 as a yellow oil (250 mg). LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.39H.sub.54N.sub.4O.sub.7:
691.87. found: 691.54.
[0814] Step 3. Preparation of 16-3: To a solution of 16-2 (250 mg,
0.36 mmol) in deoxygenated DCE (0.005 M) was added Zhan 1B catalyst
(26 mg, 0.036 mmol, Strem) and the reaction was deoxygenated for
another 10 minutes with Ar. The reaction was heated to 70.degree.
C. for 2 h. The reaction mixture was allowed to cool to rt and
concentrated. The resulting residue was directly purified by silica
gel chromatography to afford 16-3 as a yellow oil (250 mg).
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.50N.sub.4O.sub.7: 663.82. found: 663.42.
[0815] Step 4. Preparation of 16-4: To a solution of 16-3 (200 mg,
0.3 mmol) in EtOAc (10 mL) was added Pd/C (10 wt % Pd, 100 mg)
under argon. The atmosphere was replaced with hydrogen and the
reaction was stirred at rt for 1.5 h. The reaction was filtered
over Celite, washed with EtOH and concentrated to give 16-4 as an
oil (180 mg) that was used subsequently without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.52N.sub.4O.sub.7: 665.83. found: 665.36.
[0816] Step 5. Preparation of 16-5: To a solution of 16-4 (165 mg,
0.25 mmol) in DCM (5 mL) was added TFA (2 mL) and the reaction was
stirred at rt for 4 h. The solvent was removed under reduced
pressure the reaction was diluted with ethyl acetate (15 mL). The
resulting solution was washed with sat. aqueous NaHCO.sub.3 and
concentrated to afford 16-5 which was used in the next step without
further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.33H.sub.44N.sub.4O.sub.7: 609.73. found: 609.47.
[0817] Step 6. Preparation of Example 16: To a solution of 16-5 (70
mg, 0.12 mmol) and Intermediate A10 (65 mg, 0.21 mmol) in DCM (1
mL) was added DIPEA (0.08 mL, 0.46 mmol) followed by HATU (88 mg,
0.23 mmol). The reaction was stirred at room temperature for 3 h.
The reaction was diluted with EtOAc and washed with aqueous
NH.sub.4Cl and brine. The crude material was purified by reverse
phase HPLC (Gemini column, 58-98% MeCN/H.sub.2O+0.1% TFA) and
lyophilized to afford Example 16 (40 mg) as a TFA salt. Analytic
HPLC RetTime: 9.21 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.56F.sub.2N.sub.6O.sub.9S: 859.99. found: 859.60.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.28 (s, 1H), 7.76 (d,
J=9.2 Hz, 1H), 7.18 (d, J=9.2 Hz, 1H), 7.10 (s, 1H), 5.97-5.82 (m,
2H), 4.88 (m, 2H), 4.51-4.46 (m, 3H), 4.19-4.11 (m, 3H), 3.90 (s,
3H), 3.70-3.29 (m, 6H), 2.97-2.52 (m, 3H), 2.06-1.41 (m, 20H),
1.39-1.17 (m, 4H), 1.09-0.89 (m, 4H), 0.65 (m, 1H), 0.46-0.44 (m,
1H).
Example 17
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18,18-difluoro--
14-methoxy-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8-
H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b-
]quinoxaline-8-carboxamide
##STR00237## ##STR00238## ##STR00239##
[0819] Steps 1 and 2. Preparation of 17-2: A mixture of
Intermediate B4 (273 mg, 0.865 mmol), Intermediate E3 (234 mg,
0.865 mmol), and cesium carbonate (310 mg, 0.952 mmol) in MeCN (2.5
mL) was heated at 85.degree. C. for 36 hours. In an alternative
process, DMF was used as the solvent. Water (10 mL) was added and
the mixture was extracted with ethyl acetate. The organic phase was
dried over sodium sulfate, filtered and concentrated to afford
17-1, which was used subsequently without further purification or
after chromatography purification. The residue was treated with 35
equiv 4 N HCl in dioxane at rt for 2.5 hours. Upon addition of
diethyl ether, the hydrochloride salt of 17-2 precipitated. The
salt was collected by vacuum filtration and dried under reduced
pressure (375 mg). In an alternative process, the deprotection was
conducted in the presence of MSA in tBuOAc and DCM. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.23H.sub.30F.sub.2N.sub.3O.sub.4: 450.2.
found: 450.1.
[0820] Step 3. Preparation of 17-3: A mixture of 17-2 (370 mg,
0.761 mmol), Intermediate D11 (205 mg, 0.761 mmol), HATU (347 mg,
0.914 mmol) and DIPEA (0.795 mL, 4.57 mmol) in DMF (3 mL) was
stirred at rt overnight. The mixture was diluted with 100 mL water
and extracted with dichloromethane. The organic phase was dried
over sodium sulfate, filtered and concentrated. The crude product
mixture was purified by silica gel chromatography (EtOAc in
hexanes: 30%) to give 17-3 (236 mg). In an alternative process,
17-2 and Intermediate D11 were mixed with EDC and HOBT in the
presence of NMM in DMF to give 17-3. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.37H.sub.51F.sub.2N.sub.4O.sub.7: 701.4. found:
701.3.
[0821] Step 4. Preparation of 17-4: A solution of 17-3 (236 mg,
0.34 mmol) in DCE (67 mL) was deoxygenated with argon for 40
minutes. Zhan 1B catalyst (25 mg, 0.034 mmol, Strem) was added and
the reaction was heated in a 100.degree. C. oil bath for 40
minutes. Solvent was removed under reduced pressure and the residue
was purified by silica gel chromatography (EtOAc in hexanes: 5% to
65%) to give the 17-4 (229 mg). LCMS-ESI (m/z): [M-F].sup.+ calcd
for C.sub.35H.sub.46FN.sub.4O.sub.7: 653.3. found: 653.2.
[0822] Step 5. Preparation of 17-5: A solution of 17-4 (229 mg,
0.34 mmol) in 50 mL ethanol was hydrogenated at 1 atm hydrogen gas
over 220 mg of 10% wt Pd/C (wet) for 2.5 hours. Filtration through
Celite and concentration under reduced pressure gave a crude
residue of 17-5 (184 mg). In an alternative process, 17-4 was
hydrogenated at hydrogen gas in the presence of Rh. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for C.sub.35H.sub.49F.sub.2N.sub.4O.sub.7:
675.4. found: 675.3.
[0823] Step 6. Preparation of 17-6: Ester 17-5 (184 mg, 0.27 mmol)
in 2 mL DCM was treated with 1 mL TFA and stirred at rt for 3 h.
The reaction mixture was concentrated and then partitioned between
water and ethyl acetate. The organic phase was washed with water,
dried over anhydrous sodium sulfate, filtered and concentrated to
give 17-6 (153 mg). LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.31H.sub.41F.sub.2N.sub.4O.sub.7: 619.3. found: 619.2.
[0824] Step 7. Preparation of Example 17: A mixture of carboxylic
acid 17-6 (153 mg, 0.247 mmol), Intermediate A10 (90 mg, 0.297
mmol), HATU (113 mg, 0.297 mmol), DMAP (45 mg, 0.37 mmol) and DIPEA
(0.215 mL, 1.24 mmol) in DMF (1.5 mL) was stirred at rt for 40
minutes. The mixture was diluted with 2 N aqueous HCl (2 mL) and
extracted with dichloromethane. The organic phase was dried over
sodium sulfate, filtered and concentrated. The crude product
mixture was purified by silica gel chromatography (EtOAc in
hexanes: 30%-95%) to give Example 17 (95 mg). Analytic HPLC
RetTime: 8.79 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.53F.sub.4N.sub.6O.sub.9S: 869.3. found: 869.2.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.948 (br s, 1H), 7.99
(d, J=9.2 Hz, 1H), 7.29 (dd, J=8.8, 2.4 Hz, 1H), 7.09 (d, J=2.8 Hz,
1H), 6.57 (br s, 1H), 5.97 (td, J.sub.H-F=52 Hz, J=6.8 Hz, 1H),
5.92 (d, J=3.6 Hz, 1H), 5.322 (d, J=9.6 Hz, 1H), 4.42 (ap d, J=7.2
Hz, 1H), 4.40 (ap s, 1H), 4.34 (ap d, J=10 Hz, 1H), 4.08 (dd,
J=12.0, 3.6 Hz, 1H), 3.99-3.94 (m, 1H), 3.96 (s, 3H), 3.67 (m, 1H),
2.52 (m, 2H), 2.06 (m, 1H), 1.93 (m, 2H), 1.77 (m, 2H), 1.63 (m,
3H), 1.50 (s, 3H), 1.56-1.42 (m, 4H), 1.25 (m, 1H), 1.19 (t, J=7.2
Hz, 3H), 1.09 (s, 9H), 1.10-0.93 (m, 2H), 0.85 (m, 2H), 0.69 (m,
1H), 0.49 (m, 1H).
Example 18
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-9-methyl-3,6-
-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methano-
cyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-
-carboxamide
##STR00240## ##STR00241## ##STR00242##
[0826] Step 1. Preparation of 18-1: Intermediate B1 (1.94 g, 6.44
mmol) was dissolved in MeCN (30 mL) under Ar. Intermediate E1 (2.02
g, 7.4 mmol) and Cs.sub.2CO.sub.3 (7.5 mmol) were added, and the
resulting mixture was stirred for 8 h at rt. Additional
Intermediate E1 (200 mg, 0.73 mmol) and Cs.sub.2CO.sub.3 (245 mg,
0.75 mmol) were added and the reaction mixture was stirred an
additional 15 h. The reaction mixture was filtered through Celite
with EtOAc and concentrated. The resulting crude residue was
dissolved in CH.sub.2Cl.sub.2, concentrated onto 12 g silica gel,
and purified by silica gel chromatography (5% to 20% EtOAc in
hexanes) to provide 18-1 as a white foam (2.63 g). LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.24H.sub.33ClN.sub.3O.sub.6: 494.2.
found: 494.1.
[0827] Step 2. Preparation of 18-2: Substituted quinoxaline 18-1
(905 mg, 1.84 mmol) was dissolved in tert-butyl acetate (7 mL) and
CH.sub.2Cl.sub.2 (1.75 mL). MeSO.sub.3H (600 .mu.L, 9.2 mmol) was
added dropwise over 45 s, and the resulting yellow solution was
stirred at rt for 50 min. Additional MeSO.sub.3H (100 .mu.L, 1.5
mmol) was added in dropwise fashion and the reaction was stirred an
additional 10 min. The reaction mixture was transferred to a
stirred mixture of EtOAc (20 mL) and saturated aqueous NaHCO.sub.3
(30 mL). The phases were separated, and the aqueous phase was
extracted with EtOAc (20 mL). The combined organic phase was dried
over Na.sub.2SO.sub.4, filtered, and concentrated to afford amine
18-2 as a colorless residue (680 mg). LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.19H.sub.25ClN.sub.3O.sub.4: 394.2. found:
394.2.
[0828] Step 3. Preparation of 18-3: Amine 18-2 (680 mg, 1.73 mmol)
and Intermediate D1 (600 mg, 2.1 mmol) were dissolved in DMF (10
mL). DIPEA (925 .mu.L, 5.30 mmol) was added followed by HATU (880
mg, 2.3 mmol). The reaction was stirred 110 min at rt and was
diluted with saturated aqueous NaHCO.sub.3 (30 mL) and EtOAc (30
mL). The phases were separated and the organic phase was washed
with half-saturated brine (2.times.40 mL), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated to a crude residue.
Purification by silica gel chromatography (10% to 20% EtOAc in
hexanes) provided 18-3 as a colorless residue (703 mg).
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.48ClN.sub.4O.sub.7: 659.3. found: 659.4.
[0829] Step 4. Preparation of 18-4: A stirred heterogeneous mixture
of 18-3 (703 mg, 1.07 mmol), PdCl.sub.2(dppf)*CH.sub.2Cl.sub.2 (48
mg, 0.059 mmol) and potassium vinyltrifluoroborate (290 mg, 2.16
mmol) in EtOH (11 mL) was sparged with argon for 15 min.
Triethylamine (320 .mu.L, 2.3 mmol) was added and the mixture was
heated to 75.degree. C. for 70 min. The reaction mixture was cooled
to ambient temperature and was diluted with EtOAc (40 mL) and
half-saturated brine (30 mL). The phases were separated and the
organic phase was dried over Na.sub.2SO.sub.4, filtered, and
concentrated. Purification by silica gel chromatography (10% to 20%
to 30% EtOAc in hexanes) provided 18-4 as a yellow residue (490
mg). LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.51N.sub.4O.sub.7: 651.4. found: 651.3.
[0830] Step 5. Preparation of 18-5: 18-4 (490 mg, 0.179 mmol) was
dissolved in DCE (250 mL) and the solution was sparged with Ar for
15 min. Zhan 1B catalyst (66 mg, 0.090 mmol, Strem) was added as a
solution in DCE (5 mL) and the resulting solution was stirred at
85.degree. C. under Ar for 105 min. The reaction mixture was cooled
to rt and was adsorbed onto silica gel (7.5 g). Purification by
silica gel chromatography (10% to 30% EtOAc in hexanes) provided
18-5 as an amorphous residue (290 mg). LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.34H.sub.47N.sub.4O.sub.7: 623.3. found:
623.3.
[0831] Step 6: Preparation of 18-6: Olefin 18-5 (290 mg, 0.072
mmol) was dissolved in EtOAc (5.5 mL) and EtOH (5.5 mL) and the
reaction vessel was purged with Ar. Pd/C (10 wt % Pd, 92 mg) was
added in a single portion and the reaction vessel was purged twice
with H.sub.2. The reaction was stirred at rt under 1 atm H.sub.2
for 1.5 h and was filtered through a pad of Celite and concentrated
to afford a crude residue of 18-6 that was used without further
purification (LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.49N.sub.4O.sub.7: 625.4. found: 625.0.
[0832] Step 7. Preparation of 18-7: 18-6 (0.466 mmol) was dissolved
in CH.sub.2Cl.sub.2 (4.3 mL) under Ar. TMSOTf (210 .mu.L, 1.16
mmol) was added dropwise over 30 s. The reaction was stirred 65 min
and an additional portion of TMSOTf (50 .mu.L, 0.28 mmol) was
added. The reaction was stirred an additional 100 min and an
additional portion of TMSOTf (100 .mu.L, 0.55 mmol) was added. The
reaction was stirred an additional 105 min and was concentrated in
vacuo. The resulting crude residue was dissolved in
CH.sub.2Cl.sub.2 (20 mL) and 0.2 M aqueous NaOH (10 mL) was added.
The mixture was stirred for 5 min and was acidified with 1 M
aqueous HCl (20 mL). The phases were separated, and the aqueous
phase was extracted with CH.sub.2Cl.sub.2 (2.times.20 mL). The
combined organic phase was dried over MgSO.sub.4, filtered, and
concentrated to afford 18-7 as a brown solid (273 mg). LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.30H.sub.41N.sub.4O.sub.7: 569.3.
found: 568.9.
[0833] Step 8. Preparation of Example 18: To a suspension of acid
18-7 (28 mg, 0.049 mmol) and Intermediate A10 (26.5 mg, 0.087 mmol)
in MeCN (1.3 mL) was added DIPEA (55 .mu.L, 0.31 mmol). To the
resulting solution was added HATU (30.5 mg, 0.080 mmol). The
reaction was stirred at rt for 1 h and an additional portion of
Intermediate A10 (3 mg, 0.01 mmol) was added. After an additional
15 min, the reaction was diluted with EtOAc (30 mL) and 1 M aqueous
HCl (20 mL). The phases were separated and the aqueous phase was
extracted with EtOAc (30 mL). The combined organic phase was dried
over Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue. Purification by silica gel chromatography (10% to 40%
acetone in hexanes) provided an amorphous residue that was
lyophilized from water and MeCN to provide Example 18 as a white
amorphous solid (26.4 mg). Analytic HPLC RetTime: 8.42 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.53F.sub.2N.sub.6O.sub.9S: 819.4. found: 819.1.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.68 (s, 1H), 7.82 (d,
J=9.1 Hz, 1H), 7.19 (dd, J=9.1, 2.8 Hz, 1H), 7.08 (d, J=2.6 Hz,
1H), 6.86 (s, 1H), 6.14-5.70 (m, 1H), 5.65 (d, J=9.9 Hz, 1H),
5.56-5.50 (m, 1H), 4.53-4.40 (m, 3H), 4.12 (dd, J=11.9, 4.3 Hz,
1H), 3.93 (s, 3H), 3.81-3.74 (m, 1H), 3.06-2.64 (m, 4H), 2.10-1.35
(m, 13H), 1.13 (d, J=7.5 Hz, 3H), 1.09 (s, 9H), 1.04-0.65 (m, 6H),
0.52-0.41 (m, 1H).
Example 19
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-(difluoromethyl)cyclopropyl]-14-methoxy-9-methyl-3,6-dioxo-1,1-
a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa-
[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxami-
de
##STR00243##
[0835] Step 1. Preparation of Example 19: To a suspension of acid
18-7 (8.8 mg, 0.015 mmol) and Intermediate A9 (7.4 mg, 0.025 mmol)
in MeCN (0.5 mL) was added DIPEA (14 .mu.L, 0.08 mmol). To the
resulting solution was added HATU (9.1 mg, 0.024 mmol). The
reaction was stirred at rt for 1 h and an additional portion of
Intermediate A9 (5 mg, 0.02 mmol) and HATU (5 mg, 0.01 mmol) were
added. After an additional 1.5 h, the reaction was diluted with
EtOAc (30 mL), 0.2 M aqueous HCl (10 mL), and brine (10 mL). The
phases were separated and the aqueous phase was extracted with
EtOAc (30 mL). The combined organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue. Purification by silica gel chromatography (10% to 40%
acetone in hexanes) provided a residue that was lyophilized from
water and MeCN to provide Example 19 as a white amorphous solid
(8.5 mg). Analytic HPLC RetTime: 8.69 min. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.38H.sub.51F.sub.2N.sub.6O.sub.9S:
805.3. found: 805.2. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
10.12 (s, 1H), 7.83 (d, J=9.1 Hz, 1H), 7.19 (dd, J=9.1, 2.7 Hz,
1H), 7.09 (d, J=2.7 Hz, 1H), 6.77 (s, 1H), 6.25-5.76 (m, 1H), 5.57
(d, J=3.7 Hz, 1H), 5.51 (d, J=9.9 Hz, 1H), 4.49-4.37 (m, 3H), 4.13
(dd, J=12.2, 4.3 Hz, 1H), 3.94 (s, 3H), 3.79-3.72 (m, 1H),
3.01-2.69 (m, 4H), 2.13-2.06 (m, 1H), 2.01-1.22 (m, 9H), 1.14 (d,
J=7.2 Hz, 3H), 1.09 (s, 9H), 1.06-0.82 (m, 6H), 0.76-0.62 (m, 1H),
0.54-0.41 (m, 1H).
Example 20
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-{(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-ethylcyclopropyl}-14-methoxy-9-methyl-3,6-dioxo-1,1a,3,4,5,6,9-
,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1,1-
0,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00244##
[0837] Step 1. Preparation of Example 20: To a suspension of acid
18-7 (10 mg, 0.018 mmol) and Intermediate A3 (6.3 mg, 0.023 mmol)
in MeCN (0.5 mL) was added DIPEA (15 .mu.L, 0.086 mmol). To the
resulting solution was added HATU (9.0 mg, 0.024 mmol). The
reaction was stirred at rt for 2.5 h and an additional portion of
Intermediate A3 (6.5 mg, 0.024 mmol) was added. After an additional
45 min, the reaction was diluted with EtOAc (2 mL) and 1 M aqueous
HCl (1.5 mL). The phases were separated and the aqueous phase was
extracted with EtOAc (4.times.1.5 mL). The combined organic phase
was dried over Na.sub.2SO.sub.4, filtered, and concentrated to
afford a crude residue. Purification by silica gel chromatography
(20% to 25% to 30% acetone in hexanes) provided a residue that was
lyophilized from water and MeCN to provide Example 20 as a white
amorphous solid (8.0 mg). Analytic HPLC RetTime: 8.40 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.55N.sub.6O.sub.9S: 783.4. found: 783.2. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 9.98 (s, 1H), 7.83 (d, J=9.1 Hz, 1H),
7.19 (dd, J=9.1, 2.8 Hz, 1H), 7.09 (d, J=2.7 Hz, 1H), 6.42 (s, 1H),
5.57 (d, J=3.8 Hz, 1H), 5.36 (d, J=9.9 Hz, 1H), 4.48-4.34 (m, 3H),
4.11 (dd, J=11.8, 4.1 Hz, 1H), 3.94 (s, 3H), 3.79-3.72 (m, 1H),
2.98-2.68 (m, 4H), 1.95-0.80 (m, 33H), 0.76-0.61 (m, 1H), 0.53-0.41
(m, 1H).
Example 21
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-ethyl-1-{[(1-methylcycl-
opropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-9-methyl-3,6-dioxo-1,1a-
,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[-
18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamid-
e
##STR00245##
[0839] Step 1. Preparation of Example 21: To a suspension of acid
18-7 (94.9 mg, 0.167 mmol) and Intermediate A4 (74.5 mg, 0.263
mmol) in MeCN (2.5 mL) was added DIPEA (180 .mu.L, 1.0 mmol). To
the resulting solution was added HATU (9.0 mg, 0.024 mmol). The
reaction was stirred at rt for 110 min and additional portions of
Intermediate A4 (31 mg, 0.11 mmol) and DIPEA (50 .mu.L, 0.29 mmol)
were added. After an additional 40 min, the reaction was diluted
with EtOAc (30 mL), 0.2 M aqueous HCl (20 mL), and brine (10 mL).
The phases were separated and the aqueous phase was extracted with
EtOAc (20 mL). The combined organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue. Purification by silica gel chromatography (10% to 40%
acetone in hexanes) provided a residue that was lyophilized from
water and MeCN to provide Example 21 as a white amorphous solid
(102.1 mg). Analytic HPLC RetTime: 8.83 min. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.40H.sub.57N.sub.6O.sub.9S: 797.4.
found: 797.5. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.76 (s,
1H), 7.80 (d, J=9.1 Hz, 1H), 7.17 (dd, J=9.1, 2.8 Hz, 1H), 7.07 (d,
J=2.7 Hz, 1H), 6.92 (s, 1H), 5.58-5.42 (m, 2H), 4.48-4.36 (m, 3H),
4.09 (dd, J=11.8, 4.2 Hz, 1H), 3.92 (s, 3H), 3.79-3.74 (m, 1H),
2.97-2.66 (m, 4H), 1.80-0.88 (m, 33H), 0.84-0.77 (m, 1H), 0.77-0.61
(m, 2H), 0.52-0.40 (m, 1H).
Example 22
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2S)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-(2-fluoroethyl)cyclopropyl]-14-methoxy-9-methyl-3,6-dioxo-1,1a-
,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[-
18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamid-
e
##STR00246##
[0841] Step 1. Preparation of Example 22: To a suspension of acid
18-7 (30.1 mg, 0.0529 mmol) and Intermediate A5 (35 mg, 0.12 mmol)
in MeCN (0.5 mL) was added DIPEA (85 .mu.L, 0.49 mmol). To the
resulting solution was added HATU (34.5 mg, 0.0907 mmol). The
reaction was stirred at rt for 90 min and was diluted with EtOAc
(30 mL), 0.2 M aqueous HCl (20 mL), and brine (10 mL). The phases
were separated and the aqueous phase was extracted with EtOAc (30
mL). The combined organic phase was dried over Na.sub.2SO.sub.4,
filtered, and concentrated to afford a crude residue that was
dissolved in CH.sub.2Cl.sub.2 and adsorbed onto 2 g silica gel.
Purification by silica gel chromatography (15% to 55% acetone in
hexanes) provided a residue that was lyophilized from water and
MeCN to provide Example 22 as a white amorphous solid (35.5 mg).
Analytic HPLC RetTime: 8.54 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.39H.sub.54FN.sub.6O.sub.9S: 801.4. found: 801.3.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.95 (s, 1H), 7.82 (d,
J=9.1 Hz, 1H), 7.19 (dd, J=9.1, 2.8 Hz, 1H), 7.08 (d, J=2.7 Hz,
1H), 6.68 (s, 1H), 5.56 (d, J=3.9 Hz, 1H), 5.43 (d, J=9.9 Hz, 1H),
4.57-4.29 (m, 5H), 4.12 (dd, J=11.8, 4.1 Hz, 1H), 3.93 (s, 3H),
3.78-3.71 (m, 1H), 2.97-2.67 (m, 4H), 2.12-1.25 (m, 14H), 1.15 (d,
J=7.4 Hz, 3H), 1.10 (s, 9H), 1.06-0.89 (m, 4H), 0.76-0.62 (m, 1H),
0.53-0.42 (m, 1H).
Example 23
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2S)-2-(2-fluoroethyl)-1-{[(1--
methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-9-methyl-3,6--
dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanoc-
yclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8--
carboxamide
##STR00247##
[0843] Step 1. Preparation of Example 23: To a suspension of acid
18-7 (30.5 mg, 0.0536 mmol) and Intermediate A6 (24.8 mg, 0.0824
mmol) in MeCN (0.5 mL) was added DIPEA (60 .mu.L, 0.34 mmol). To
the resulting solution was added HATU (32.3 mg, 0.0850 mmol). The
reaction was stirred at rt for 75 min and an additional portion of
Intermediate A6 (9 mg, 0.03 mmol) was added. After an additional 75
min the reaction was diluted with EtOAc (30 mL), 0.2 M aqueous HCl
(20 mL), and brine (10 mL). The phases were separated and the
aqueous phase was extracted with EtOAc (30 mL). The combined
organic phase was dried over Na.sub.2SO.sub.4, filtered, and
concentrated to afford a crude residue that was dissolved in
CH.sub.2Cl.sub.2 and adsorbed onto 2 g silica gel. Purification by
silica gel chromatography (15% to 55% acetone in hexanes) provided
a residue that was lyophilized from water and MeCN to provide
Example 23 as a white amorphous solid (37.1 mg). Analytic HPLC
RetTime: 8.64 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.56FN.sub.6O.sub.9S: 815.4. found: 815.6. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 9.63 (s, 1H), 7.83 (d, J=9.1 Hz, 1H),
7.20 (dd, J=9.1, 2.8 Hz, 1H), 7.10 (d, J=2.7 Hz, 1H), 6.75 (s, 1H),
5.56 (d, J=3.9 Hz, 1H), 5.50 (d, J=10.0 Hz, 1H), 4.56-4.34 (m, 5H),
4.13 (dd, J=11.8, 4.2 Hz, 1H), 3.95 (s, 3H), 3.82-3.75 (m, 1H),
2.98-2.70 (m, 4H), 2.07-2.00 (m, 1H), 2.00-1.93 (m, 1H), 1.88-1.44
(m, 12H), 1.32-1.26 (m, 1H), 1.17 (d, J=7.4 Hz, 3H), 1.12 (d,
J=10.6 Hz, 9H), 1.07-0.83 (m, 4H), 0.81-0.65 (m, 2H), 0.52-0.44 (m,
1H).
Example 24
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2S)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-(2,2-difluoroethyl)cyclopropyl]-14-methoxy-9-methyl-3,6-dioxo--
1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopr-
opa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carbox-
amide
##STR00248##
[0845] Step 1. Preparation of Example 24: To a suspension of acid
18-7 (30.2 mg, 0.0531 mmol) and Intermediate A7 (25.9 mg, 0.0850
mmol) in MeCN (0.5 mL) was added DIPEA (60 .mu.L, 0.34 mmol). To
the resulting solution was added HATU (32 mg, 0.084 mmol). The
reaction was stirred at rt for 75 min and an additional portion of
Intermediate A7 (3.0 mg, 0.0098 mmol) was added. After an
additional 30 min the reaction was diluted with EtOAc (30 mL), 0.2
M aqueous HCl (20 mL), and brine (10 mL). The phases were separated
and the aqueous phase was extracted with EtOAc (30 mL). The
combined organic phase was dried over Na.sub.2SO.sub.4, filtered,
and concentrated to afford a crude residue that was dissolved in
CH.sub.2Cl.sub.2 and adsorbed onto 2 g silica gel. Purification by
silica gel chromatography (15% to 55% acetone in hexanes) provided
a residue that was lyophilized from water and MeCN to provide
Example 24 as a white amorphous solid (35.5 mg). Analytic HPLC
RetTime: 8.62 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.53F.sub.2N.sub.6O.sub.9S: 819.4. found: 819.2.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.99 (s, 1H), 7.82 (d,
J=9.1 Hz, 1H), 7.19 (dd, J=9.1, 2.8 Hz, 1H), 7.08 (d, J=2.7 Hz,
1H), 6.69 (s, 1H), 5.99-5.64 (m, 1H), 5.56 (d, J=3.9 Hz, 1H), 5.40
(d, J=10.0 Hz, 1H), 4.47-4.39 (m, 3H), 4.14-4.08 (m, 1H), 3.93 (s,
3H), 3.78-3.72 (m, 1H), 2.96-2.67 (m, 4H), 2.29-2.16 (m, 2H),
1.83-1.24 (m, 12H), 1.15 (d, J=7.4 Hz, 3H), 1.09 (s, 9H), 1.05-0.82
(m, 4H), 0.74-0.63 (m, 1H), 0.53-0.42 (m, 1H).
Example 25
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2S)-2-(2,2-difluoroethyl)-1-{-
[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-9-methyl--
3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-meth-
anocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxalin-
e-8-carboxamide
##STR00249##
[0847] Step 1. Preparation of Example 25: To a suspension of acid
18-7 (30.3 mg, 0.0532 mmol) and Intermediate A8 (28.3 mg, 0.0887
mmol) in MeCN (0.5 mL) was added DIPEA (60 .mu.L, 0.34 mmol). To
the resulting solution was added HATU (32.4 mg, 0.0852 mmol). The
reaction was stirred at rt for 2.5 h and was diluted with EtOAc (30
mL), 0.2 M aqueous HCl (20 mL), and brine (10 mL). The phases were
separated and the aqueous phase was extracted with EtOAc (30 mL).
The combined organic phase was dried over Na.sub.2SO.sub.4,
filtered, and concentrated to afford a crude residue that was
dissolved in CH.sub.2Cl.sub.2 and adsorbed onto 2 g silica gel.
Purification by silica gel chromatography (15% to 55% acetone in
hexanes) provided a residue that was lyophilized from water and
MeCN to provide Example 25 as a white amorphous solid (33.9 mg).
Analytic HPLC RetTime: 8.66 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.40H.sub.55F.sub.2N.sub.6O.sub.9S: 833.4. found: 833.4.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.62 (s, 1H), 7.82 (d,
J=9.1 Hz, 1H), 7.18 (dd, J=9.1, 2.8 Hz, 1H), 7.08 (d, J=2.7 Hz,
1H), 6.64 (s, 1H), 6.04-5.66 (m, 1H), 5.54 (d, J=4.0 Hz, 1H), 5.47
(d, J=10.0 Hz, 1H), 4.50-4.38 (m, 3H), 4.11 (dd, J=11.8, 4.2 Hz,
1H), 3.93 (s, 3H), 3.82-3.71 (m, 1H), 2.98-2.68 (m, 4H), 2.27-2.11
(m, 2H), 1.96-1.41 (m, 12H), 1.32 (dd, J=9.6, 5.4 Hz, 1H), 1.15 (d,
J=7.4 Hz, 3H), 1.10 (s, 9H), 1.05-0.64 (m, 6H), 0.51-0.42 (m,
1H).
Example 26
Preparation of
(1R,4S,4aR,8S,11S,12S,13R,25aR)-8-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsu-
lfonyl)carbamoyl]-2-(difluoromethyl)cyclopropyl]-17-methoxy-12-methyl-6,9--
dioxo-2,3,4,4a,6,7,8,9,12,13,21,22,23,24,25,25a-hexadecahydro-1H,11H-1,4:1-
0,13-dimethanoquinoxalino[2,3-k][1,10,3,6]benzodioxadiazacyclononadecine-1-
1-carboxamide
##STR00250## ##STR00251## ##STR00252##
[0849] Step 1. Preparation of 26-2: To a solution of 26-1 (311 mg,
0.710 mmol; prepared similarly to 18-1 of Example 18 substituting
Intermediate B2 for Intermediate B1 in step 1) in dioxane (1.8 mL)
was added 4 M HCl in dioxane (1.8 mL, 7.2 mmol). The reaction was
stirred for 15.5 h at rt and was then concentrated under reduced
pressure to give 26-2 as a white amorphous solid that was used
without further purification in the following step. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for C.sub.16H.sub.19ClN.sub.3O.sub.4:
352.1. found: 352.2.
[0850] Steps 2 and 3. Preparation of diastereomeric mixture 26-3
and 26-4: Amine hydrochloride 26-2 (0.710 mmol) was dissolved along
with 1:1 mixture of Intermediate mixture D9 and D10 (266 mg, 0.788
mmol) and DIPEA (600 .mu.L, 3.4 mmol) in DMF (4.5 mL). HATU (360
mg, 0.95 mmol) was added in one portion. The reaction was stirred
1.75 h at rt and was diluted with saturated aqueous NaHCO.sub.3 (20
mL), water (10 mL) and EtOAc (30 mL). The phases were separated and
the organic phase was washed twice with a mixture of water (30 mL)
and brine (5 mL). The organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated to a crude residue that
was purified by silica gel chromatography (10% to 30% EtOAc in
hexanes) to provide a colorless residue (380 mg; LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.35H.sub.48ClN.sub.4O.sub.7: 671.3.
found: 671.6). A stirred heterogeneous mixture of this residue,
PdCl.sub.2(dppf)*CH.sub.2Cl.sub.2 (35 mg, 0.043 mmol) and potassium
vinyltrifluoroborate (156 mg, 1.16 mmol) in EtOH (7 mL) was sparged
with argon for several minutes. Triethylamine (170 .mu.L, 1.2 mmol)
was added and the mixture was heated to 70.degree. C. for 55 min.
The reaction mixture was cooled to ambient temperature, diluted
with EtOAc (40 mL), and washed with water (30 mL). The organics
were dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated to afford a residue that was purified by silica gel
chromatography (15% to 30% EtOAc in hexanes) to afford
diastereomeric mixture 26-3 and 26-4 as a yellow residue (277 mg).
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.51N.sub.4O.sub.7: 663.4. found: 663.3.
[0851] Step 4. Preparation of 26-5: Diastereomeric mixture 26-3 and
26-4 (277 mg, 0.419 mmol) was dissolved in DCE (140 mL) and the
solution was sparged with Ar for 15 min. Zhan 1B catalyst (37 mg,
0.050 mmol, Strem) was added and the resulting solution was stirred
at 85.degree. C. under Ar for 1.5 h. The reaction mixture was then
concentrated and purified by silica gel chromatography (20% to 50%
EtOAc in hexanes) to afford 26-5 as an amorphous residue (105 mg).
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.47N.sub.4O.sub.7: 635.3. found: 635.3.
[0852] Steps 5 and 6. Preparation of 26-6: To a solution of 26-5
(105 mg, 0.165 mmol) in 1:1 EtOAc:EtOH (4 mL) was added Pd/C (10 wt
% Pd, 43 mg). The reaction vessel was purged twice with H.sub.2 and
was stirred at rt under 1 atm H.sub.2 for 1 h. The reaction mixture
was filtered through a pad of Celite and concentrated to afford a
crude residue (106 mg; LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.49N.sub.4O.sub.7: 637.4. found: 637.3). This residue
was then dissolved in THF (0.8 mL). MeOH (0.4 mL), water (0.4 mL)
and LiOH*H.sub.2O (67 mg, 1.6 mmol) were added and the mixture was
stirred at 45.degree. C. for 14.5 h. The reaction was quenched
dropwise with 1 N aqueous HCl (1.3 mL) and was diluted with
CH.sub.2Cl.sub.2 (30 mL) and 1 N aqueous HCl (20 mL). The phases
were separated, and the aqueous phase was extracted with
CH.sub.2Cl.sub.2 (30 mL). The combined organic phase was dried over
MgSO.sub.4, filtered and concentrated to afford 26-6 as a residue
(93.8 mg) that was used directly in Step 7. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.34H.sub.47N.sub.4O.sub.7: 623.3. found:
623.3.
[0853] Step 7. Preparation of Example 26: To a suspension of acid
26-6 (93.8 mg, 0.151 mmol) and Intermediate A9 (58 mg, 0.20 mmol)
in MeCN was added DIPEA (120 .mu.L, 0.69 mmol). To the resulting
solution was added HATU (73.5 mg, 0.193 mmol). The reaction was
stirred at rt for 100 min and an additional portion of Intermediate
A9 (6 mg, 0.02 mmol) was added. After an additional 30 min,
additional Intermediate A9 (9 mg, 0.03 mmol), HATU (9 mg, 0.02
mmol) and DIPEA (10 .mu.L, 0.06 mmol) were added. The reaction was
stirred for an additional 50 min and was diluted with EtOAc (25
mL), 0.2 M aqueous HCl (20 mL) and brine (10 mL). The phases were
separated and the aqueous phase was extracted with EtOAc (25 mL).
The combined organic phase was dried over Na.sub.2SO.sub.4,
filtered, and concentrated to afford a crude residue. Purification
by silica gel chromatography (25% to 40% acetone in hexanes)
provided an amorphous residue that was lyophilized from water and
MeCN to provide Example 26 as a white amorphous solid (113 mg).
Analytic HPLC RetTime: 9.19 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.42H.sub.57F.sub.2N.sub.6O.sub.9S: 859.4. found:
859.2. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.02 (s, 1H),
7.80 (d, J=9.1 Hz, 1H), 7.21-7.15 (m, 2H), 7.07 (d, J=2.7 Hz, 1H),
6.13-5.79 (m, 1H), 5.63 (d, J=10.1 Hz, 1H), 5.50-5.45 (m, 1H), 4.51
(d, J=10.1 Hz, 1H), 4.44 (d, J=7.4 Hz, 1H), 4.25 (s, 1H), 4.18-4.12
(m, 2H), 3.93 (s, 3H), 3.02-2.77 (m, 3H), 2.66-2.57 (m, 1H),
2.18-0.90 (m, 36H).
Example 27
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[-
(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-16-methoxy-11-methyl--
5,8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-10H-9,12-
-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quino-
xaline-10-carboxamide
##STR00253## ##STR00254##
[0855] Step 1. Preparation of 27-1: Amine hydrochloride 26-2 (217
mg, 0.504 mmol), was treated with BEP (207 mg, 0.756 mmol),
Intermediate 05 (283 mg, 0.909 mmol), EtOAc (9 mL), NMP (1 mL) and
DIPEA (0.44 m, 2.5 mmol), then heated to 50.degree. C. After 1.5 h,
the reaction mixture was diluted with EtOAc. The organic solution
was washed successively with saturated aqueous NaHCO.sub.3 and
brine, then dried over MgSO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by silica gel
chromatography (9% to 40% EtOAc/Hex) to afford amide 27-1 (235 mg).
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.52ClN.sub.4O.sub.7: 687.35. found: 688.13.
[0856] Step 2. Preparation of 27-2: Amide 27-1 (235 mg, 0.342 mmol)
was treated with potassium vinyltrifluoroborate (69 mg, 0.513
mmol), Pd(dppf)Cl.sub.2*DCM (28 mg, 0.0342 mmol), EtOH (3.4 mL) and
TEA (0.072 mL, 0.513 mmol), then heated to reflux. After 50 min,
the reaction mixture was diluted with EtOAc and washed with
H.sub.2O and brine. The organics were dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by silica gel chromatography (9% to 40% EtOAc/Hex) to
afford vinyl quinoxaline 27-2 (219 mg). LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.38H.sub.55N.sub.4O.sub.7: 679.41. found:
679.49.
[0857] Steps 3 and 4. Preparation of 27-3: Vinyl quinoxaline 27-2
(219 mg, 0.323 mmol) was suspended in DCE (65 mL) and treated with
Zhan 1B catalyst (41 mg, 0.065 mmol, Strem). The suspension was
deoxygenated with bubbling N.sub.2 for 17 min, then heated to
reflux for 90 min. The reaction mixture was then filtered over
Celite and concentrated under reduced pressure. The crude residue
was purified by silica gel chromatography (15% to 50% EtOAc/Hex) to
afford the desired macrocycle (165 mg; LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.36H.sub.51N.sub.4O.sub.7: 651.38.
found: 651.40). The macrocyclic product of step 3 was dissolved in
EtOH (10 mL) and EtOAc (2 mL) and treated with 10 wt % Pd/C (95
mg). Hydrogen from a balloon was bubbled through the suspension for
1 min and the mixture was stirred under H.sub.2 (1 atm) for an
additional 1.5 h. The reaction mixture was filtered over Celite and
concentrated under reduced pressure to afford the desired
macrocycle 27-3 which was carried on without further purification.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.53N.sub.4O.sub.7: 653.39. found: 653.32.
[0858] Step 5. Preparation of 27-4: The crude product of step 4 was
dissolved in DCM and treated with TMSOTf (0.23 mL, 1.3 mmol). After
stirring at rt for 1 h 15 min, the reaction mixture was
concentrated under reduced pressure. The residue was redissolved in
DCM and added by pipette to a separatory funnel containing 1 M
aqueous NaOH. The mixture was agitated for 1 min, then acidified to
pH 1-2 with 10% aqueous HCl. The aqueous layer was extracted three
times with DCM and combined organics dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The crude
material was purified by silica gel chromatography to afford
carboxylic acid 27-4 (119 mg). LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.32H.sub.45N.sub.4O.sub.7: 597.33. found: 597.40.
[0859] Step 6. Preparation of Example 27: Carboxylic acid 27-4 (105
mg, 0.177 mmol) and Intermediate A10 (65 mg, 0.212 mmol) were
treated with TBTU (68 mg, 0.212 mmol), DMAP (26 mg, 0.212 mmol),
DCM (1.8 mL) and DIPEA (0.31 mL, 1.8 mmol). The reaction mixture
was stirred at rt for 30 min, then more amine A10 (40 mg, 0.131
mmol) was added and the reaction mixture was heated to reflux.
After an additional 1.25 h, the mixture was concentrated under
reduced pressure. The crude residue was purified by HPLC to afford
Example 27 (80 mg) in approximately 90% purity as a TFA salt.
Analytic HPLC RetTime: 9.06 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S: 847.39. found: 847.69.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.23 (s, 1H), 7.87-7.72
(m, 1H), 7.31-7.14 (m, 2H), 5.84 (td, J=55.6, 6.5 Hz, 1H), 5.58 (d,
J=22.6 Hz, 1H), 4.94-4.81 (m, 1H), 4.37 (d, J=15.8 Hz, 1H),
4.29-4.10 (m, 2H), 3.94 (s, 3H), 3.01 (ddd, J=15.1, 9.9, 5.3 Hz,
1H), 2.84 (p, J=7.4 Hz, 1H), 2.75 (ddd, J=13.3, 10.2, 6.0 Hz, 1H),
2.03 (d, J=9.0 Hz, 2H), 1.97-1.74 (m, 4H), 1.73-1.55 (m, 6H), 1.53
(s, 3H), 1.48-1.21 (m, 8H), 1.19-1.02 (m, 14H), 0.99-0.80 (m,
2H).
Example 28
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-(difluoromethyl)cyclopropyl]-16-methoxy-11-methyl-5,8-dioxo--
1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-10H-9,12-methanocy-
clopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-10--
carboxamide
##STR00255##
[0861] Step 1. Carboxylic acid 27-4 (20 mg, 0.034 mmol) and
Intermediate A9 (35 mg, 0.12 mmol) were treated with TBTU (22 mg,
0.067 mmol), DMAP (8 mg, 0.07 mmol), DCM (1 mL) and DIPEA (0.117
mL, 0.674 mmol). The reaction mixture was stirred at rt for 15 h,
then concentrated under reduced pressure. The crude residue was
purified by HPLC to afford Example 28 (22 mg) in approximately 90%
purity as a TFA salt. Analytic HPLC RetTime: 8.90 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.55F.sub.2N.sub.6O.sub.9S: 833.37. found: 833.61.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.23 (s, 1H), 7.79 (d,
J=8.8 Hz, 1H), 7.34-7.10 (m, 2H), 5.86 (td, J=55.8, 6.5 Hz, 1H),
5.61 (s, 1H), 4.54 (t, J=9.7 Hz, 1H), 4.36 (d, J=16.5 Hz, 1H),
4.28-4.07 (m, 2H), 3.95 (d, J=17.8 Hz, 3H), 3.08-2.91 (m, 2H),
2.90-2.79 (m, 1H), 2.73 (ddd, J=13.3, 10.3, 6.0 Hz, 1H), 2.04 (s,
2H), 1.97-1.74 (m, 4H), 1.64 (ddd, J=18.7, 11.6, 4.0 Hz, 4H),
1.49-1.19 (m, 11H), 1.18-0.94 (m, 14H), 0.94-0.80 (m, 1H).
Example 29
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-3,6-dioxo-9--
propyl-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methano-
cyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-
-carboxamide
##STR00256## ##STR00257## ##STR00258##
[0863] Step 1. Preparation of 29-1: To a solution of Intermediate
B5 (188 mg, 0.57 mmol) and Intermediate E1 (233 mg, 0.86 mmol) in
MeCN (2.85 mL) was added cesium carbonate (280 mg, 9.18 mmol) at rt
under an argon atmosphere. After 19 h, the reaction mixture was
then filtered through a pad of Celite and the filtrate concentrated
in vacuo. The crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes gradient) to afford
substituted quinoxaline 29-1 (240 mg) as a colorless oil. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.26H.sub.37ClN.sub.3O.sub.6:
522.2. found: 522.3.
[0864] Step 2. Preparation of 29-2: To a solution 29-1 (240 mg,
0.46 mmol) in dioxane (1 mL) was added 4 M hydrochloric acid in
dioxane (4 mL, 1 mmol) and the reaction stirred at rt. After 15 h,
the reaction mixture was concentrated in vacuo to afford amine
hydrochloride 29-2 (200 mg) as an off white solid, which was used
directly in the next step without further purification. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.21H.sub.29ClN.sub.3O.sub.4:
422.2. found: 422.2.
[0865] Step 3. Preparation of 29-3: To a solution of 29-2 (200 mg,
0.46 mmol) and Intermediate D1 (170 mg, 0.51 mmol) in MeCN (2.3 mL)
was added HATU (192 mg, 0.51 mmol) followed by DIPEA (400 .mu.L,
2.30 mmol) at rt under and argon atmosphere. After 1.5 h, the
reaction mixture was concentrated in vacuo, and the crude residue
was purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford amide 29-3 (67 mg) as a
colorless oil. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.52ClN.sub.4O.sub.7: 687.3. found: 687.5.
[0866] Step 4. Preparation of 29-4: To a solution of 29-3 (67 mg,
98 .mu.mol), TEA (20 .mu.L, 150 .mu.mol) and potassium
vinyltrifluoroborate (19.7 mg, 150 .mu.mol) in EtOH (500 .mu.L) was
added PdCl.sub.2(dppf) (8 mg, 9.8 .mu.mol). The reaction mixture
was deoxygenated with argon for 10 min and was heated to 78.degree.
C. After 40 min, the reaction mixture was allowed to cool to rt and
was concentrated in vacuo. The crude residue was purified by silica
gel chromatography (0-100% ethyl acetate/hexanes gradient) to
afford vinyl quinoxaline 29-4 (40.2 mg) as a colorless oil.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.55N.sub.4O.sub.7: 679.4. found: 679.6.
[0867] Step 5. Preparation of 29-5: To a solution of 29-4 (40 mg,
59 .mu.mol) in DCE (11.8 mL) was added Zhan 1B catalyst (4 mg, 6
.mu.mol, Strem) and the reaction mixture was degassed for 10
minutes with argon. The reaction mixture was then heated to
100.degree. C. After 1 h, the reaction mixture was allowed to cool
to rt and was concentrated in vacuo. The crude residue was purified
by silica gel chromatography (0-100% ethyl acetate/hexanes
gradient) to afford macrocycle 29-5 (31 mg) as a light yellow oil.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.51N.sub.4O.sub.7: 651.4. found: 651.5.
[0868] Step 6. Preparation of 29-6: To a solution of macrocycle
29-5 (31 mg, 47 .mu.mol) in ethanol (500 .mu.L) was added Pd/C (10
wt %, 5 mg, 5 .mu.mol) at rt under an argon atmosphere. The
reaction vessel was evacuated and refilled with 1 atm hydrogen gas
(3.times.) and the reaction mixture was stirred vigorously at rt.
After 1 h, the reaction mixture was diluted with ethyl acetate (10
mL) and was filtered through a pad of Celite with ethyl acetate
washings (3.times.5 mL). The filtrate was concentrated in vacuo to
afford macrocycle 29-6 (31 mg), which was used directly in the next
step without further purification. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.36H.sub.53N.sub.4O.sub.7: 653.4. found: 653.5.
[0869] Step 7. Preparation of 29-7: To a solution of 29-6 (31 mg,
47 .mu.mol) in DCM (0.5 mL) was added TMSOTf (44 .mu.L, 0.25 mmol)
at rt under an argon atmosphere. After 25 min, the reaction mixture
was concentrated in vacuo and was azeotropically dried from toluene
(2.times.2 mL) to afford carboxylic acid 29-7 (35 mg) as a yellow
oil, which was used directly in the next step without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.45N.sub.4O.sub.7: 597.3. found: 597.4.
[0870] Step 8. Preparation of Example 29: To a solution of 29-7 (35
mg, 49 .mu.mol) and Intermediate A10 (22 mg, 74 .mu.mol) in MeCN
(245 .mu.L) was added HATU (28 mg, 74 .mu.mol) followed by DIPEA
(43 .mu.L, 250 .mu.mol) at rt under an argon atmosphere. After 3 h,
the reaction mixture was concentrated in vacuo, was purified by
preparatory HPLC (Gemini 5u C18 110 .ANG. column, 5-100%
MeCN/H.sub.2O, 0.1% trifluoroacetic acid modifier) and was
lyophilized to afford Example 29 (22.3 mg) as a white powder TFA
salt. Analytic HPLC RetTime: 8.81 min. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.41H.sub.56F.sub.2N.sub.6O.sub.9S: 847.4. found:
847.5. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.83 (d, J=9.4 Hz,
1H), 7.93 (d, J=9.1 Hz, 1H), 7.36 (d, J=9.1 Hz, 1H), 7.21 (d,
J=11.0 Hz, 1H), 7.14 (s, 1H), 5.97 (td, J.sub.H-F=55 Hz, J=7.2 Hz,
1H), 5.84 (br s, 1H), 5.41 (d, J=9.4 Hz, 1H), 4.66-4.34 (m, 3H),
4.13 (app d, J=11.8 Hz, 1H), 4.08 (s, 1H), 3.97 (s, 3H), 3.78-3.71
(m, 1H), 3.09-2.65 (m, 5H), 2.14-2.04 (m, 1H), 1.87-1.34 (m, 8H),
1.52 (s, 3H), 1.12 (s, 9H), 1.08-0.84 (m, 10H), 0.76-0.62 (m, 1H),
0.50 (dd, J=12.6, 6.6 Hz, 1H).
Example 30
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-9-(2-methylp-
ropyl)-3,6-dioxo-1,1a,3,4,5,6,9,
10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1,1-
0,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00259## ##STR00260## ##STR00261##
[0872] Step 1. Preparation of 30-1: A mixture of Intermediate B6
(139 mg, 0.405 mmol), Intermediate E1 (170 mg, 0.625 mmol), and
cesium carbonate (203 mg, 0.623 mmol) in 3.3 mL of acetonitrile was
stirred at room temperature under argon overnight. Reaction mixture
was filtered over Celite, washing with ethyl acetate, and filtrate
was concentrated under reduced pressure. The resulting residue was
purified by silica gel chromatography (0-30% ethyl acetate in
hexanes) to yield 30-1 (170 mg) as a clear film. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.27H.sub.39ClN.sub.3O.sub.6: 536.24.
found: 536.31.
[0873] Step 2. Preparation of 30-2: A solution of hydrogen chloride
in dioxane (4.0 M, 0.16 mL, 0.64 mmol) was added to a solution of
30-1 (168 mg, 0.314 mmol) in 3.3 mL of dioxane at room temperature.
After thirty minutes, an additional 4 equivalents of HCl was added
and mixture was stirred overnight. An additional 25 equivalents of
HCl was then added. After thirty minutes, an additional 19
equivalents of HCl was added. After one hour, an additional 29
equivalents of HCl was added. After thirty minutes, reaction
mixture was concentrated under reduced pressure to yield 30-2 (148
mg, 85% purity), which was used in the next step without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.22H.sub.31ClN.sub.3O.sub.4: 436.19. found: 436.25.
[0874] Step 3. Preparation of 30-3: HATU (144 mg, 0.379 mmol,
Oakwood) and DIPEA (0.28 mL, 1.58 mmol) were added to a mixture of
30-2 (148 mg, 0.315 mmol) and Intermediate D1 (99 mg, 0.348 mmol)
in 3.5 mL of DMF under argon. After stirring overnight, reaction
mixture was poured into water and extracted with ethyl acetate
(3.times.). Combined organics were washed with water and brine,
dried (MgSO.sub.4), filtered, and concentrated under reduced
pressure. The resulting residue was purified by silica gel
chromatography (0-50% ethyl acetate in hexanes) to yield 30-3 (136
mg) as a white solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.54ClN.sub.4O.sub.7: 701.36. found: 701.47.
[0875] Step 4. Preparation of 30-4:
Pd(dppf).sub.2Cl.sub.2*CH.sub.2Cl.sub.2 (35 mg, 0.043 mmol) was
added to a degassed mixture of 30-3 (135 mg, 0.193 mmol), potassium
vinyltrifluoroborate (41 mg, 0.306 mmol), and triethylamine (0.040
mL, 0.289 mmol) in 2.1 mL of ethanol at room temperature. Reaction
mixture was heated at 78.degree. C. under argon for 45 minutes.
After cooling to room temperature, reaction mixture was poured into
water and extracted with ethyl acetate (three times). Combined
organics were washed with water and brine, dried (MgSO.sub.4),
filtered, and concentrated under reduced pressure to yield 30-4
(133 mg), which was used in the next step without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.57N.sub.4O.sub.7: 693.41. found: 693.48.
[0876] Step 5. Preparation of 30-5: A mixture of 30-4 (133 mg,
0.192 mmol) and Zhan 1B catalyst (16 mg, 0.022 mmol, Strem) in 38
mL of DCE was deoxygenated under argon for 25 minutes. The mixture
was then heated at 95.degree. C. for 50 minutes. After cooling to
room temperature, reaction mixture was concentrated under reduced
pressure. The resulting residue was purified by silica gel
chromatography (0-50% ethyl acetate in hexanes) to yield 30-5 (70
mg) as a light yellow film. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.53N.sub.4O.sub.7: 665.38. found: 665.50.
[0877] Step 6. Preparation of 30-6: Palladium on carbon (10 wt %
Pd, 22 mg, 0.0208 mmol) was added to a solution of 30-5 (69 mg,
0.104 mmol) in 3 mL of ethanol. Mixture was then stirred under an
atmosphere of hydrogen for 1 hour and then was filtered over
Celite, washing with ethyl acetate. Filtrate was concentrated under
reduced pressure to yield 30-6 (64 mg) as a light yellow-brown
solid film, which was used in the next step without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.55N.sub.4O.sub.7: 667.40. found: 667.43.
[0878] Step 7. Preparation of 30-7: TMSOTf (0.050 mL, 0.274 mmol)
was added dropwise to a solution of 30-6 (30 mg, 0.045 mmol) in 1.2
mL of dichloromethane under argon at room temperature. After 45
minutes, reaction mixture was concentrated under reduced pressure.
The resulting film was taken up in 5 mL of toluene and concentrated
under reduced pressure. This process was repeated a second time to
yield 30-7 (27 mg), which was used in the next step without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.33H.sub.47N.sub.4O.sub.7: 611.34. found: 611.41.
[0879] Step 8. Preparation of Example 30: HATU (28 mg, 0.074 mmol,
Oakwood) and DIPEA (0.050 mL, 0.281 mmol) were added to a mixture
of 30-7 (27 mg, 0.045 mmol) and Intermediate A10 (22 mg, 0.072
mmol) in 2.2 mL of acetonitrile under argon. After stirring
overnight, reaction mixture was poured into water and extracted
with ethyl acetate (3.times.). Combined organics were washed with
water and brine, dried (MgSO.sub.4), filtered, and concentrated
under reduced pressure. The resulting residue was purified by
silica gel chromatography (0-50% ethyl acetate in hexanes) and
reverse phase prep HPLC (15-100% acetonitrile in water, with 0.1%
trifluoroacetic acid buffer) to yield the trifluoroacetic acid salt
of Example 30 (18 mg) as a light yellow solid, after
lyophilization. Analytic HPLC RetTime: 8.96 min. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.42H.sub.59F.sub.2N.sub.6O.sub.9S:
861.40. found: 861.30. .sup.1H NMR (400 MHz, CD.sub.3OD): .delta.
9.17 (s, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.23 (dd, J=8.8, 2.8 Hz, 1H),
7.14 (d, J=2.8 Hz, 1H), 5.81 (td, J.sub.H-F=56 Hz, J=7.6 Hz, 1H);
5.77 (d, J=3.2 Hz, 1H), 4.55 (d, J=7.2 Hz, 1H), 4.39 (t, J=5.6 Hz,
2H), 4.16 (dd, J=11.8, 4 Hz, 1H), 3.91 (s, 3H), 3.79-3.71 (m, 1H),
2.98-2.90 (m, 1H), 2.84 (dd, J=12.6, 4.8 Hz, 1H), 2.79-2.72 (m,
1H), 2.06-1.91 (m, 3H), 1.77 (m, 3H), 1.64-1.44 (m, 6H), 1.51 (s,
3H), 1.44-1.32 (m, 3H), 1.15-1.07 (m, 1H), 1.10 (s, 9H), 1.06-0.96
(m, 3H), 1.04-1.01 (m, 6H), 0.93-0.89 (m, 2H), 0.79-0.68 (m, 1H),
0.52-0.47 (m, 1H).
Example 31
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-9-cyclopropyl-N-[(1R,2R)-2-(difluoro-
methyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methox-
y-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-me-
thanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxal-
ine-8-carboxamide
##STR00262## ##STR00263## ##STR00264##
[0881] Step 1. Preparation of 31-1: An unpurified sample of
Intermediate B3 was treated with Intermediate E1 (217 mg, 0.797
mmol), MeCN (5.7 mL) and Cs.sub.2CO.sub.3 (371 mg, 1.14 mmol).
After stirring at rt for 17 h, the reaction mixture was filtered
over Celite and concentrated under reduced pressure. The crude
residue was purified by silica gel chromatography (20% to 40%
EtOAc/Hex) to afford quinoxaline 31-1 (143 mg). LCMS-ESI (m/z):
[M-Boc+2H].sup.+ calcd for C.sub.18H.sub.21ClN.sub.3O.sub.4:
378.12. found: 378.59.
[0882] Step 2. Preparation of 31-2: Quinoxaline 31-1 (143 mg, 0.299
mmol) was dissolved in DCM (10 mL) and treated with HCl (4.0 M in
dioxane, 5 mL, 20.0 mmol). After stirring for 2 h at rt, the
reaction mixture was concentrated and the crude 31-2 was carried on
without further purification.
[0883] Step 3. Preparation of 31-3: The crude amine hydrochloride
31-2 was treated with BEP (115 mg, 0.419 mmol), Intermediate D1
(120 mg, 0.423 mmol), EtOAc (9 mL), NMP (1 mL) and DIPEA (0.37 mL,
2.1 mmol), then heated to 50.degree. C. After 1.5 h, the reaction
mixture was diluted with Et.sub.2O. The organic solution was washed
successively with saturated aqueous NaHCO.sub.3 and brine, then
dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by silica gel chromatography
(15% to 30% EtOAc/Hex) to afford amide 31-3 (166 mg). LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.33H.sub.44ClN.sub.4O.sub.7:
643.29. found: 643.48.
[0884] Step 4. Preparation of 31-4: Amide 31-3 (166 mg, 0.258 mmol)
was treated with potassium vinyltrifluoroborate (52 mg, 0.387
mmol), Pd(dppf)Cl.sub.2*DCM (21 mg, 0.0258 mmol), EtOH (2.6 mL) and
TEA (0.054 mL), then heated to reflux. After 50 min, the reaction
mixture was diluted with EtOAc and washed with H.sub.2O and brine.
The organics were dried over MgSO.sub.4, filtered and concentrated
under reduced pressure. The residue was purified by silica gel
chromatography (15% to 40% EtOAc/Hex) to afford vinyl quinoxaline
31-4 (145 mg). LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.47N.sub.4O.sub.7: 635.34. found: 635.58.
[0885] Steps 5 and 6. Preparation of 31-5: Vinyl quinoxaline 31-4
(145 mg, 0.228 mmol) was suspended in DCE (46 mL) and treated with
Zhan 1B catalyst (33 mg, 0.0456 mmol, Strem). The suspension was
deoxygenated with bubbling N.sub.2 for 22 min, then heated to
reflux for 50 min. The reaction mixture was then filtered over
Celite and concentrated under reduced pressure. The crude residue
was purified by silica gel chromatography (25% to 35% EtOAc/Hex) to
afford the desired macrocycle (54 mg; LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.33H.sub.43N.sub.4O.sub.7: 607.31.
found: 607.67). The macrocyclic product of step 5 was dissolved in
EtOH (10 mL) and treated with 10% Pd/C (45 mg). Hydrogen from a
balloon was bubbled through the suspension for 1 min and
hydrogenation (1 atm) was continued for an additional 1.5 h. The
reaction mixture was filtered over Celite and concentrated under
reduced pressure to afford the desired macrocycle 31-5 which was
carried on without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.33H.sub.45N.sub.4O.sub.7: 609.33.
found: 609.95.
[0886] Step 7. Preparation of 31-6: The crude product 31-5 was
dissolved in THF and treated with LiOH (1.0 M in H.sub.2O, 5 mL, 5
mmol). After stirring at rt for 3 d, the reaction mixture was
heated to reflux for 20 h. The mixture was then poured into
H.sub.2O and acidified to pH .about.1-2 with 10% HCl. The aqueous
layer was extracted three times with DCM. The combined organics
were dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The crude material was purified by silica gel
chromatography (80% to 100% EtOAc/Hex) to afford carboxylic acid
31-6 (24 mg). LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.43N.sub.4O.sub.7: 595.31. found: 595.12.
[0887] Step 8. Preparation of Example 31: Carboxylic acid 31-6 (24
mg, 0.040 mmol) and Intermediate A10 (25 mg, 0.081 mmol) were
treated with TBTU (23 mg, 0.081 mmol), DMAP (10 mg, 0.081 mmol),
DCM (2 mL) and DIPEA (0.070 mL, 0.40 mmol). The reaction mixture
was stirred at rt for 15 h then concentrated under reduced
pressure. The crude residue was purified by HPLC to afford Example
31 (13 mg, 34%) in approximately 90% purity as a TFA salt. Analytic
HPLC RetTime: 8.92 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.55F.sub.2N.sub.6O.sub.9S: 845.37. found: 845.67.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.13 (s, 1H), 7.79 (d,
J=9.1 Hz, 1H), 7.23 (dd, J=9.1, 2.7 Hz, 1H), 7.13 (d, J=2.7 Hz,
1H), 6.05-5.65 (m, 2H), 4.55 (d, J=7.0 Hz, 1H), 4.47 (d, J=11.7 Hz,
2H), 4.27 (dd, J=12.0, 3.7 Hz, 1H), 3.94 (s, 3H), 3.78 (dd, J=6.8,
2.8 Hz, 1H), 2.99-2.86 (m, 1H), 2.80 (td, J=13.2, 4.1 Hz, 1H), 1.98
(d, J=28.8 Hz, 2H), 1.92-1.67 (m, 4H), 1.65-1.41 (m, 10H), 1.33 (d,
J=27.7 Hz, 3H), 1.20-1.06 (m, 9H), 1.04-0.84 (m, 6H), 0.82-0.62 (m,
3H), 0.61-0.41 (m, 2H), 0.06 (dd, J=9.2, 4.9 Hz, 1H).
Example 32
Preparation of
(1aR,5S,8S,9S,10R,22aR)-9-benzyl-5-tert-butyl-N-[(1R,2R)-2-(difluoromethy-
l)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-3,6-
-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methano-
cyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-
-carboxamide
##STR00265## ##STR00266## ##STR00267##
[0889] Step 1. Preparation of 32-1: To a solution of Intermediate
B7 (390 mg, 1.00 mmol) and Intermediate E1 (272 mg, 1.00 mmol) in
MeCN (5 mL) was added cesium carbonate (390 mg, 1.00 mmol) at rt
under an argon atmosphere. After 24 h, the reaction mixture was
diluted with ethyl acetate (50 mL). The resulting mixture was
washed with saturated aqueous sodium bicarbonate solution (50 mL)
and brine (50 mL), was dried over anhydrous sodium sulfate, and was
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes gradient) to afford
quinoxaline 32-1 (550 mg) as a colorless oil. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.30H.sub.37ClN.sub.3O.sub.6: 570.2.
found: 570.2.
[0890] Step 2. Preparation of 32-2: To a solution of 32-1 (549 mg,
0.96 mmol) in dioxane (2 mL) was added 4 M hydrochloric acid in
dioxane (2 mL, 1 mmol) and the reaction was stirred at rt. After 24
h, the reaction mixture was concentrated in vacuo to afford amine
hydrochloride 32-2 (461 mg) as an off white solid, which was used
directly in the next step without further purification.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.25H.sub.29ClN.sub.3O.sub.4: 470.2. found: 470.2.
[0891] Step 3. Preparation of 32-3: To a solution of 32-2 (461 mg,
0.96 mmol) and Intermediate D1 (369 mg, 1.10 mmol) in MeCN (5 mL)
was added HATU (418 mg, 1.10 mmol) followed by DIPEA (869 .mu.L,
5.00 mmol) at rt under and argon atmosphere. After 24 h, the
reaction mixture was concentrated in vacuo, and the crude residue
was purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford 32-3 (202.6 mg) as a colorless
oil. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.52ClN.sub.4O.sub.7: 735.3. found: 735.4.
[0892] Step 4. Preparation of 32-4: To a solution of 32-3 (202 mg,
276 .mu.mol), TEA (56 .mu.L, 414 .mu.mol) and potassium
vinyltrifluoroborate (56 mg, 414 .mu.mol) in EtOH (2.76 mL) was
added PdCl.sub.2(dppf) (22.5 mg, 27.6 .mu.mol). The reaction
mixture was degassed with argon for 10 min and was heated to
78.degree. C. After 1 h, the reaction mixture was allowed to cool
to rt and was concentrated in vacuo. The crude residue was purified
by silica gel chromatography (0-100% ethyl acetate/hexanes
gradient) to afford 32-4 (163 mg) as a yellow oil. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for C.sub.42H.sub.55N.sub.4O.sub.7: 727.4.
found: 727.5.
[0893] Step 5. Preparation of 32-5: To a solution of 32-4 (163 mg,
220 .mu.mol) in DCE (44 mL) was added Zhan 1B catalyst (16 mg, 22
.mu.mol, Strem) and the reaction mixture was degassed for 10
minutes with argon. The reaction mixture was then heated to
100.degree. C. After 45 min, the reaction mixture was allowed to
cool to rt and was concentrated in vacuo. The crude residue was
purified by silica gel chromatography (0-100% ethyl acetate/hexanes
gradient) to afford 32-5 (125 mg) as a light yellow oil. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.40H.sub.51N.sub.4O.sub.7: 699.4.
found: 699.4.
[0894] Step 6. Preparation of 32-6: To a solution of macrocycle
32-5 (124 mg, 178 .mu.mol) in ethanol (890 .mu.L) was added Pd/C
(10 wt % Pd, 19 mg, 18 .mu.mol) at rt under an argon atmosphere.
The reaction vessel was evacuated and refilled with hydrogen gas
(3.times.) and the reaction mixture was stirred vigorously at rt
under 1 atm H.sub.2. After 2.5 h, the reaction mixture was diluted
with ethyl acetate (5 mL) and was filtered through a pad of Celite
with ethyl acetate washings (3.times.5 mL). The filtrate was
concentrated in vacuo to afford 32-6 (139 mg), which was used
directly in the next step without further purification. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.40H.sub.53N.sub.4O.sub.7: 701.4.
found: 701.5.
[0895] Steps 7 and 8. Preparation of Example 32: To a solution of
32-6 (124 mg, 178 .mu.mol) in DCM (3 mL) was added TFA (2 mL) at rt
under an argon atmosphere. After 3 h, the reaction mixture was
concentrated in vacuo and was azeotropically dried from toluene
(2.times.2 mL) to afford the desired carboxylic acid as a yellow
oil, which was used directly in the next step without further
purification. (126 mg; LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.45N.sub.4O.sub.7: 645.3. found: 645.4). To a solution
of this carboxylic acid (120 mg, 178 .mu.mol) and Intermediate A10
(119 mg, 392 .mu.mol) in MeCN (1 mL) was added HATU (151 mg, 392
.mu.mol) followed by DIPEA (155 .mu.L, 890 .mu.mol) at rt under an
argon atmosphere. After 30 min, the reaction mixture was
concentrated in vacuo, and the crude residue was purified by silica
gel chromatography (0-100% ethyl acetate/hexanes gradient). The
fractions containing the desired product were combined, were
repurified by preparatory HPLC (Gemini 5u C18 110 .ANG. column,
5-100% MeCN/H.sub.2O, 0.1% trifluoroacetic acid modifier) and were
lyophilized to afford the TFA salt of Example 32 (23 mg) as a white
powder. Analytic HPLC RetTime: 8.81 min. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.45H.sub.57F.sub.2N.sub.6O.sub.9S:
895.4. found: 895.6. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.24
(s, 1H), 7.73 (d, J=9.1 Hz, 1H), 7.47-7.27 (m, 4H), 7.21-7.12 (m,
1H), 6.65 (d, J=2.9 Hz, 1H), 5.83 (td, J.sub.H-F=55 Hz, J=7.2 Hz,
1H), 5.77 (br s, 1H), 4.63 (d, J=6.9 Hz, 2H), 4.50-4.28 (m, 3H),
3.93 (s, 2H), 3.79-3.71 (m, 1H), 3.11-2.99 (m, 1H), 2.97-2.85 (m,
1H), 2.82-2.61 (m, 3H), 1.92 (br s, 2H), 1.82-1.70 (m, 2H),
1.63-1.44 (m, 4H), 1.52 (s, 3H), 1.15 (s, 9H), 1.04 (br s, 2H),
1.02-0.96 (m, 2H), 0.95-0.88 (m, 4H), 0.78-0.66 (m, 1H), 0.56-0.46
(m, 1H).
Example 33
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1S,2R)-1-[(cyclopropyl-
sulfonyl)carbamoyl]-2-(difluoromethyl)cyclopropyl]-15-methoxy-10-methyl-4,-
7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadecahydro-1H,9H-8-
,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6]dioxadiazacyc-
lononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00268## ##STR00269## ##STR00270##
[0897] Steps 1 and 2. Preparation of diastereomeric mixture 33-1
and 33-2: Quinoxaline 18-2 (220 mg, 0.56 mmol) was dissolved along
with 1:1 diastereomer Intermediate mixture D12 and D13 (208 mg,
0.643 mmol) in MeCN (5 mL). DIPEA (280 .mu.L, 1.6 mmol) and HATU
(360 mg, 0.95 mmol) were added, and the reaction was stirred for
1.25 h at rt. The reaction was then diluted with EtOAc (30 mL),
saturated aqueous NaHCO.sub.3 (15 mL), H.sub.2O (10 mL), and brine
(10 mL). The phases were separated and the aqueous phase was
extracted with EtOAc (30 mL). The organic phase was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated to a crude
residue that was dissolved in CH.sub.2Cl.sub.2 and adsorbed onto
silica gel (5 g). Purification by silica gel chromatography (10% to
30% EtOAc in hexanes) provided a white foam (352 mg; LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.37H.sub.52ClN.sub.4O.sub.7:
699.4. found: 699.1). A stirred heterogeneous mixture of this
residue, PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 (30.7 mg, 0.0376 mmol)
and potassium vinyltrifluoroborate (135 mg, 1.01 mmol) in EtOH (5
mL) was sparged with argon for several minutes. Triethylamine (160
.mu.L, 1.1 mmol) was added and the mixture was heated to 75.degree.
C. for 1 h. The reaction mixture was cooled to ambient temperature
and was diluted with EtOAc (30 mL), H.sub.2O (15 mL) and brine (15
mL). The phases were separated, and the aqueous phase was extracted
with EtOAc (30 mL). The organics were dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated to afford a crude
residue that was dissolved in CH.sub.2Cl.sub.2 and adsorbed onto
silica gel (3 g). Purification by silica gel chromatography (10% to
40% EtOAc in hexanes) produced inseparable mixture of 33-1 and 33-2
as a yellow residue (258 mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.55N.sub.4O.sub.7: 691.4. found: 691.7.
[0898] Step 3: Preparation of 33-3: Diastereomeric mixture 33-1 and
33-2 (258 mg, 0.373 mmol) was dissolved in DCE (125 mL) and the
solution was sparged with Ar for 10 min. Zhan 1B catalyst (41 mg,
0.056 mmol, Strem) was added as a solution in DCE (3.3 mL) and the
resulting solution was stirred at 85.degree. C. under Ar for 105
min. The reaction mixture was then concentrated onto 5 g silica gel
and was purified by silica gel chromatography (0% to 25% EtOAc in
hexanes) to afford macrocycle 33-3 as an amorphous residue (81.9
mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.51N.sub.4O.sub.7: 663.4. found: 663.3.
[0899] Steps 4 and 5: Preparation of 33-4: To a solution of 33-3
(81.9 mg, 0.124 mmol) in 1:1 EtOAc:EtOH (4 mL) was added Pd/C (10
wt % Pd, 19 mg). The reaction vessel was purged twice with H.sub.2
and was stirred at rt under 1 atm H.sub.2 for 2.5 h. The reaction
mixture was filtered through a pad of Celite and concentrated to
afford a crude residue. This residue was dissolved in
CH.sub.2Cl.sub.2 (1.2 mL) and TMSOTf (90 .mu.L, 0.50 mmol) was
added. The mixture was stirred at rt for 4.5 h. The reaction was
then concentrated in vacuo and dissolved in CH.sub.2Cl.sub.2 (5
mL). 0.2 M aqueous NaOH (5 mL) was added and the biphasic mixture
was stirred at rt for 5 min. The mixture was then acidified with 1
M aqueous HCl (20 mL) and was diluted with CH.sub.2Cl.sub.2 (20
mL). The phases were separated and the aqueous phase was extracted
with CH.sub.2Cl.sub.2 (2.times.20 mL). The combined organic phase
was dried over MgSO.sub.4, filtered, and concentrated to afford
33-4 as a crude residue (76.1 mg). LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.33H.sub.45N.sub.4O.sub.7: 609.3. found: 608.9.
[0900] Step 6: Preparation of Example 33: To a suspension of acid
33-4 (43 mg, 0.072 mmol) and Intermediate A9 (40.9 mg, 0.14 mmol)
in MeCN (800 .mu.L) was added DIPEA (100 .mu.L, 0.57 mmol). HATU
(37 mg, 0.097 mmol) was added to the resulting solution, and the
reaction was stirred at rt for 15 h. The reaction was then diluted
with EtOAc (20 mL), 0.2 M aqueous HCl (10 mL) and brine (10 mL).
The phases were separated and the aqueous phase was extracted with
EtOAc (20 mL). The combined organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue. This residue was dissolved in CH.sub.2Cl.sub.2 and was
concentrated onto 2 g silica gel. Purification by silica gel
chromatography (15% to 55% acetone in hexanes) provided an
amorphous residue that was lyophilized from water and MeCN to
provide Example 33 as a white amorphous solid (29.6 mg). Analytic
HPLC RetTime: 9.07 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.55F.sub.2N.sub.6O.sub.9S: 845.4. found: 845.2.
.sup.1H NMR (400 MHz, CDCl.sub.3) b 10.21 (s, 1H), 7.82 (d, J=9.1
Hz, 1H), 7.19 (dd, J=9.1, 2.7 Hz, 1H), 7.09 (d, J=2.7 Hz, 1H), 6.79
(s, 1H), 6.21-5.76 (m, 1H), 5.65 (d, J=3.9 Hz, 1H), 5.29 (d, J=9.7
Hz, 1H), 4.99 (d, J=7.5 Hz, 1H), 4.47-4.29 (m, 4H), 4.16-4.09 (m,
1H), 3.93 (s, 3H), 2.99-2.85 (m, 2H), 2.80-2.64 (m, 2H), 2.24-2.16
(m, 1H), 2.13-2.05 (m, 1H), 2.01-0.95 (m, 29H), 0.56-0.45 (m, 1H),
0.45-0.35 (m, 1H).
Example 34
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-{(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-ethylcyclopropyl}-9-ethyl-18,18-difluoro-14-methoxy-3,6-dioxo--
1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopr-
opa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carbox-
amide
##STR00271##
[0902] Example 34 was prepared in a similar fashion to Example 17,
substituting Intermediate A3 for Intermediate A10 in Step 7.
Example 34 was isolated (5.7 mg) in approximately 95% purity.
Analytic HPLC RetTime: 8.81 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.40H.sub.55F.sub.2N.sub.6O.sub.9S: 833.4. found: 833.25.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.027 (br s, 1H), 7.98
(d, J=8.8 Hz, 1H), 7.29 (dd, J=9.2, 2.8 Hz, 1H), 7.09 (d, J=2.8 Hz,
1H), 6.32 (br s, 1H), 5.92 (d, J=3.6 Hz, 1H), 5.30 (d, J=10.0 Hz,
1H), 4.42-4.33 (m, 3H), 4.08 (dd, J=11.6, 4.0 Hz, 1H), 3.96 (s,
3H), 3.65 (m, 1H), 2.93 (m, 1H), 2.51 (m, 2H), 2.02 (m, 1H),
1.86-1.40 (m, 11H) 1.34-1.14 (m, 7H), 1.09 (s, 9H), 1.10-0.82 (m,
6H), 0.72 (m, 1H), 0.48 (m, 1H).
Example 35
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2S)-2-(2,2-difluoroethyl)-1-{-
[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18,18-difluo-
ro-14-methoxy-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydr-
o-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,1-
2-b]quinoxaline-8-carboxamide
##STR00272##
[0904] Example 35 was prepared in a similar fashion to Example 17,
substituting Intermediate A8 for Intermediate A10 in Step 7.
Example 35 was isolated (12.8 mg) in approximately 90% purity.
Analytic HPLC RetTime: 8.78 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.41H.sub.55F.sub.4N.sub.6O.sub.9S: 883.4. found: 883.2.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.69 (br s, 1H), 7.98 (d,
J=9.2 Hz, 1H), 7.29 (dd, J=9.2, 2.8 Hz, 1H), 7.09 (d, J=2.8 Hz,
1H), 6.53 (br s, 1H), 5.91 (d, J=4.0 Hz, 1H), 5.84 (tt,
J.sub.H-F=56 Hz, J=3.6 Hz, 1H), 5.33 (d, J=6.4 Hz, 1H), 4.43 (m,
2H), 4.34 (ap d, J=9.6 Hz, 1H), 4.08 (dd, J=11.6, 4.0 Hz, 1H), 3.96
(s, 3H), 3.99-3.94 (m, 1H), 3.68 (m, 1H), 2.58-2.52 (m, 3H), 2.20
(m, 2H), 1.82-1.58 (m, 7H) 1.54-1.40 (m, 5H), 1.36-1.18 (m, 6H),
1.09 (s, 9H), 1.10-1.00 (m, 1H), 0.85 (m, 2H), 0.69 (m, 1H), 0.49
(m, 1H).
Example 36
Preparation of
(1aR,5S,8S,9S,10R,21aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-3,6--
dioxo-1a,3,4,5,6,9,10,17b,18,18a,19,20,21,21a-tetradecahydro-1H,8H-7,10-me-
thanodicyclopropa[13,14:18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]-
quinoxaline-8-carboxamide
##STR00273## ##STR00274##
[0906] Step 1. Preparation of 36-1: To a solution of
trimethylsulfoxonium iodide (72 mg, 0.32 mmol) in DMSO/THF (1:1, 2
mL) was added sodium hydride (60%, 12 mg, 0.32 mmol) and stirred at
rt for 2 h. Macrocycle 1-5 (103 mg, 0.16 mmol) was added dropwise
in THF (3 mL). The mixture was heated to 65.degree. C. and stirred
for 16 h. After cooling to rt, the mixture was diluted with
EtOAc/H.sub.2O, extracted with EtOAc, dried over anhydrous
MgSO.sub.4, and concentrated in vacuo. The residue was purified by
silica gel chromatography (0-25% EtOAc/hexanes) to give 36-1 (27
mg) as a residue. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.51N.sub.4O.sub.7: 651.38. found: 651.52.
[0907] Step 2. Preparation of 36-2: To a solution of 36-1 (26 mg,
0.04 mmol) in DCM (1 mL) was added TMSOTf (0.036 mL, 0.2 mmol) and
stirred at rt for 2 h. The reaction was pipetted into stirring 1 N
NaOH (2 mL). After 10 min, the mixture was diluted with DCM and
acidified to pH 3 with 1 N aqueous HCl. Following extraction of the
aqueous layer with DCM, the combined organics were dried over
anhydrous MgSO.sub.4 and concentrated in vacuo. The residue was
purified by silica gel chromatography (0-10% EtOAc/MeOH) to give
36-2 (24 mg) as a residue that was used without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.43N.sub.4O.sub.7: 595.31. found: 595.43.
[0908] Step 3. Preparation of Example 36: To a solution of 36-2 (24
mg, 0.041 mmol), Intermediate A10 (16 mg, 0.053 mmol), TBTU (19 mg,
0.06 mmol) and DMAP (8 mg, 0.06 mmol) in DCM (2 mL) was added DIPEA
(0.021 mL, 0.12 mmol) and the reaction was stirred at rt for 16 h.
Additional Intermediate A10 (16 mg, 0.053 mmol), TBTU (19 mg, 0.06
mmol), DMAP (8 mg, 0.06 mmol), and DIPEA (0.021 mL, 0.12 mmol) were
added and the reaction was stirred at rt for 4 h. The reaction was
quenched with water, diluted with EtOAc, washed with sat. aqueous
NaHCO.sub.3, brine, dried over anhydrous MgSO.sub.4 and
concentrated in vacuo. The crude material was purified by reverse
phase HPLC (Gemini, 45-85% MeCN/H.sub.2O+0.1% TFA) and lyophilized
to give Example 36 (3 mg) as a TFA salt. Analytic HPLC RetTime:
9.06 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.55F.sub.2N.sub.6O.sub.9S: 845.37. found: 845.43.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.31 (s, 1H), 7.72 (d,
J=10 Hz, 1H), 7.20-7.17 (m, 2H), 5.60-5.82 (m, 2H), 5.51 (s, 1H),
4.72 (d, J=7.2 Hz, 1H), 4.43 (d, J=11.6 Hz, 1H), 4.31 (s, 1H),
4.26-4.22 (dd, J=11.6, 4 Hz, 1H), 3.94 (s, 3H), 3.78 (m, 1H), 2.60
(m, 1H), 2.27 (m, 1H), 2.04 (s, 3H), 1.68 (m, 3H), 1.59 (m, 2H),
1.54-1.15 (m, 11H), 1.09 (s, 9H), 0.95-0.86 (m, 8H), 0.47 (m,
1H).
Example 37
Preparation of
(1R,4S,4aR,8S,11S,12S,13R,25aR)-8-tert-butyl-N-[(1R,2R)-2-(difluoromethyl-
)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-12-ethyl-17-met-
hoxy-6,9-dioxo-2,3,4,4a,6,7,8,9,12,13,21,22,23,24,25,25a-hexadecahydro-1H,-
11H-1,4:10,13-dimethanoquinoxalino[2,3-k][1,10,3,6]benzodioxadiazacyclonon-
adecine-11-carboxamide
##STR00275##
[0910] Step 1. Preparation of Example 37: To a solution of 13-6 (76
mg, 0.12 mmol), Intermediate A10 (44 mg, 0.14 mmol), HATU (55 mg,
0.14 mmol) and DMAP (21 mg, 0.18 mmol) in DMF (2 mL) was added
DIPEA (0.11 mL, 0.6 mmol) and the reaction was stirred at rt for 16
h. Additional Intermediate A10 (44 mg, 0.14 mmol), HATU (55 mg,
0.14 mmol), DMAP (21 mg, 0.18 mmol), followed by DIPEA (0.11 mL,
0.6 mmol) was added and the reaction was stirred at 40.degree. C.
for 50 h. The reaction was quenched with water, diluted with EtOAc,
washed with sat. aqueous NaHCO.sub.3, brine, dried over anhydrous
MgSO.sub.4 and concentrated in vacuo. The crude material was
purified by reverse phase HPLC (Gemini, 45-85% MeCN/H.sub.2O+0.1%
TFA) and lyophilized to give Example 37 (30 mg) as a TFA salt.
Analytic HPLC RetTime: 9.44 min. LCMS-ESI (m/z): [M+H]*calcd for
C.sub.44H.sub.61F.sub.2N.sub.6O.sub.9S: 887.42. found: 887.50.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.24 (s, 1H), 7.76 (d,
J=9.2 Hz, 1H), 7.20 (dd, J=8.8, 2.4 Hz, 1H), 7.12 (m, 1H),
5.95-5.66 (m, 2H), 5.43 (s, 1H), 4.51 (d, J=7.6 Hz, 1H), 4.41 (s,
1H), 4.20-4.10 (m, 2H), 3.88 (s, 3H), 2.94-2.88 (m, 1H), 2.73-2.63
(m, 2H), 2.11 (br, 2H), 2.02-0.83 (m, 41H).
Example 38
Preparation of
(1aR,5S,8S,9S,10R,22aR)--N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-methylcyclo-
propyl)sulfonyl]carbamoyl}cyclopropyl]-14-m
ethoxy-9-methyl-5-(1-methylcyclopentyl)-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19-
,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dio-
xadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00276## ##STR00277##
[0912] Step 1. Preparation of 38-1: Amine 18-2 (192 mg, 0.487 mmol)
was treated with BEP (246 mg, 0.898 mmol), Intermediate D14 (278
mg, 0.898 mmol), EtOAc (9 mL), NMP (1 mL) and DIPEA (0.42 mL, 2.4
mmol), then heated to 50.degree. C. After 1 h, the reaction mixture
was diluted with EtOAc. The organic solution was washed
successively with sat. aqueous NaHCO.sub.3 and brine, then dried
over MgSO.sub.4, filtered and concentrated in vacuo. The residue
was purified by silica gel chromatography (15% to 35% EtOAc/Hex) to
afford amide 38-1 (264 mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.50ClN.sub.4O.sub.7: 685.34. found: 685.82.
[0913] Step 2. Preparation of 38-2: Amide 38-1 (264 mg, 0.385 mmol)
was treated with potassium vinyltrifluoroborate (82 mg, 0.615
mmol), Pd(dppf)Cl.sub.2*DCM (33 mg, 0.041 mmol), EtOH (4.0 mL) and
TEA (0.086 mL, 0.62 mmol), then heated to reflux. After 55 min, the
reaction mixture was diluted with EtOAc and washed with H.sub.2O
and brine. The organics were dried over anhydrous MgSO.sub.4,
filtered and concentrated in vacuo. The residue was purified by
silica gel chromatography (15% to 30% EtOAc/Hex) to afford vinyl
quinoxaline 38-2 (168 mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.53N.sub.4O.sub.7: 677.39. found: 677.38.
[0914] Steps 3 and 4. Preparation of 38-3: Vinyl quinoxaline 38-2
(225 mg, 0.332 mmol) was suspended in DCE (66 mL) and treated with
Zhan 1B catalyst (42 mg, 0.067 mmol, Strem). The suspension was
degassed with bubbling N.sub.2 for 28 min, then heated to reflux
for 90 min. The reaction mixture was then filtered over Celite and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (15% to 30% EtOAc/Hex) to afford the desired
macrocycle (168 mg; LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49N.sub.4O.sub.7: 649.36. found: 649.33). The
macrocycle was dissolved in EtOH (25 mL) and EtOAc (5 mL) and
treated with Pd/C (10 wt % Pd, 95 mg). Hydrogen from a balloon was
bubbled through the suspension for 1 min the reaction was stirred
under an H.sub.2 atmosphere for an additional 1.5 h. Upon
completion, the reaction mixture was filtered over Celite and
concentrated in vacuo to afford the desired macrocycle 38-3 which
was carried on without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.36H.sub.51N.sub.4O.sub.7: 651.38.
found: 651.42.
[0915] Step 5. Preparation of 38-4: Unpurified 38-3 from the
previous step was dissolved in DCM (10 mL) and treated with TMSOTf
(0.23 mL, 1.3 mmol). After stirring at rt for 1 h 15 min, the
reaction mixture was concentrated in vacuo. The residue was
redissolved in DCM and pipetted into 1 M aqueous NaOH. The mixture
was agitated for 1 min, then acidified to pH 1-2 with 10% aqueous
HCl. The aqueous layer was extracted three times with DCM and
combined organics dried over anhydrous MgSO.sub.4, filtered and
concentrated in vacuo. The crude material was purified by silica
gel chromatography (0% to 20% MeOH/EtOAc) to afford carboxylic acid
38-4 (131 mg). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.43N.sub.4O.sub.7: 595.31. found: 595.29.
[0916] Step 6. Preparation of Example 38: Carboxylic acid 38-4 (131
mg, 0.220 mmol) and Intermediate A10 (81 mg, 0.264 mmol) were
treated with TBTU (85 mg, 0.264 mmol), DMAP (32 mg, 0.264 mmol),
DCM (2.6 mL) and DIPEA (0.38 mL, 2.2 mmol). The reaction mixture
was stirred at rt for 14 h, then concentrated under reduced
pressure. The crude residue was purified by HPLC to afford Example
38 (74 mg) in approximately 90% purity as a TFA salt. Analytic HPLC
RetTime: 8.93 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.55F.sub.2N.sub.6O.sub.9S: 845.37. found: 845.57.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.12 (s, 1H), 7.77 (d,
J=9.2 Hz, 1H), 7.20 (dd, J=9.0 Hz, 2.7 Hz, 1H), 7.16 (d, J=2.8 Hz,
1H), 5.81 (td, J=55.9, 6.6 Hz, 1H), 5.59 (d, J=3.5 Hz, 1H), 4.52
(d, J=6.8 Hz, 1H), 4.50 (s, 1H), 4.40 (d, J=12.0 Hz, 1H), 4.18 (dd,
J=11.9 Hz, 3.9 Hz, 1H), 3.93 (s, 3H), 3.74 (m, 1H), 2.97-2.90 (m,
1H), 2.85-2.75 (m, 2H), 2.01 (m, 2H), 1.85-1.41 (m, 21H), 1.12 (s,
3H), 1.08 (d, J=7.4 Hz, 3H), 0.96 (m, 2H), 0.91 (t, J=4.3 Hz, 2H),
0.70 (m, 1H), 0.48 (m, 1H).
Example 39
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[-
(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-1-ethyl-16-methoxy-3a-
-methyl-5,8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro--
10H-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-
-b]quinoxaline-10-carboxamide
##STR00278## ##STR00279## ##STR00280##
[0918] Step 1. Preparation of 39-1: Quinoxaline ether 1-1 (588.7
mg, 1.159 mmol) was dissolved in TFA (5 mL). The solution was
stirred at room temperature for 3 h. The TFA was removed in vacuo
to give the TFA salt of 39-1 (631.2 mg) as a colorless powder.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.16H.sub.19C.sub.1N.sub.3O.sub.4: 352.1. found: 352.1.
[0919] Step 2. Preparation of 39-2: The TFA salt of 39-1 (631.2 mg,
1.159 mmol) was dissolved in CH.sub.2Cl.sub.2/MeOH (3 mL/3 mL). To
the solution was added a solution of TMSCHN.sub.2 (2 M hexane, 3
mL, 5.177 mmol) at rt. The solution was stirred for 30 min to
produce a suspension that was filtered through a fritted glass
funnel to remove solids. The filtrate was concentrated in vacuo to
afford a residue that was purified by silica gel chromatography
(100% ethyl acetate) to produce methyl ester 39-2 (213.0 mg) as
colorless crystals. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.17H.sub.21ClN.sub.3O.sub.4: 366.1. found: 366.1.
[0920] Step 3. Preparation of 39-3: Intermediate D7 (191.2 mg,
0.587 mmol) and methyl ester 39-2 (414.1 mg, 1.132 mmol) were
treated with HATU (860.0 mg, 2.264 mmol) and DIPEA (0.59 mL, 3.396
mmol) in DMF (8 mL) at rt for 4 h. The reaction was quenched with
H.sub.2O (50 mL) and extracted with EtOAc (50 mL three times). The
combined organics were washed with brine (50 mL) and dried over
anhydrous Na.sub.2SO.sub.4. After removal of drying agent by
filtration, the solvent was removed in vacuo. The residue was
purified by silica gel chromatography (20% ethyl acetate in
hexanes) to give the desired amide 39-3 (573.9 mg) as colorless
oil. LCMS-ESI.sup.+ (m/z): [M+Na].sup.+ calcd for
C.sub.33H.sub.49ClN.sub.4NaO.sub.7: 695.3. found: 695.3.
[0921] Step 4. Preparation of 39-4: Amide 39-3 (573.9 mg, 0.8524
mmol), potassium trifluorovinylborate (171.3 mg, 1.279 mmol) and
PdCl.sub.2dppf.CH.sub.2Cl.sub.2 (62.4 mg, 0.085 mmol) were treated
with Et.sub.3N (0.18 mL, 1.279 mmol) in EtOH (8 mL) under a
nitrogen atmosphere and gently refluxed for 30 min. The reaction
was diluted with PhMe (30 mL) and the solvent was removed in vacuo.
The residue was purified by silica gel chromatography (20% ethyl
acetate in hexanes) to give the desired vinyl quinoxaline 39-4
(542.0 mg, 0.8152 mmol) as an orange foam. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.37H.sub.52N.sub.4NaO.sub.7: 687.4.
found: 687.3.
[0922] Step 5. Preparation of 39-5: The vinyl quinoxaline 39-4
(542.0 mg, 0.8152 mmol) was treated with Zhan 1b catalyst (59.8 mg,
0.08 mmol, Strem) in DCE (41 mL). The mixture was heated at
80.degree. C. for 1 h. Additional Zhan 1b catalyst (59.8 mg, 0.08
mmol, Strem) was added and the mixture to heat at 80.degree. C. for
an additional 30 min. The solvent was removed in vacuo and the
residue purified by silica gel chromatography (20% ethyl acetate in
hexanes) to produce macrocycle 39-5 (401.0 mg, 0.6297 mmol) as an
orange oil. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.49N.sub.4O.sub.7: 637.4. found: 637.3.
[0923] Step 6. Preparation of 39-6: Macrocycle 39-5 (401.0 mg,
0.6297 mmol) was taken up in 1,4-dioxane (15 mL) and treated with
Pd/C (10% wt Pd, 200.0 mg) and MgO (200.0 mg) stirred under an
atmosphere of hydrogen. The mixture was stirred at rt for 1 h. The
reaction mixture was filtered through Celite (5 g) using EtOAc (80
mL). The solvent was removed in vacuo to give macrocycle 39-6
(425.3 mg) as a pale orange oil. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.35H.sub.51N.sub.4O.sub.7: 639.4. found: 639.3.
[0924] Step 7. Preparation of 39-7: Macrocycle 39-6 (74.8 mg, 0.110
mmol) was treated with 2 M aqueous LiOH aqueous solution (1.6 mL,
3.15 mmol) in MeOH/THF (4 mL/4 mL) at rt for 8 h, 50.degree. C. for
2 h and then 60.degree. C. for 3 h. The mixture was cooled to
0.degree. C. using ice-water bath. To the mixture was added brine
(30 mL). The whole was extracted with CH.sub.2Cl.sub.2 (30 mL three
times). The organic layer was washed with brine (30 mL) and dried
over anhydrous Na.sub.2SO.sub.4. After removal of the drying agent
by filtration, the solvent was removed in vacuo to give carboxylic
acid 39-7 (370.6 mg, 0.5932 mmol) as a colorless oil.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.49N.sub.4O.sub.7: 625.4. found: 625.3.
[0925] Step 8. Preparation of Example 39: Carboxylic acid 39-7
(100.0 mg, 0.1601 mmol) and Intermediate A10 (73.2 mg, 0.2401 mmol)
were treated with HATU (91.3 mg, 0.2401 mmol) and DIPEA (0.14 mL,
0.8005 mmol) in DMF (3 mL) at rt for 5 h. The reaction was quenched
with H.sub.2O (30 mL) and extracted with EtOAc (30 mL three times).
The organic layer was washed with brine (30 mL) and dried over
anhydrous Na.sub.2SO.sub.4. After removal of the drying agent by
filtration, the solvent was removed in vacuo. The residue was
purified by silica gel chromatography (25 to 100% ethyl acetate in
hexanes). Fractions containing desired product were concentrated in
vacuo and the residue further purified by super critical fluid
column chromatography (DAICEL Chiralpak IC 10.times.250 mm, 18.9
mL/min, 35% MeOH, 15 atm, 40.degree. C.) to give Example 39 (80.5
mg, 0.0920 mmol, 57%) as a colorless powder. Analytic HPLC RetTime:
9.35 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.43H.sub.61F.sub.2N.sub.6O.sub.9S: 875.4. found: 875.4.
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.81 (d, J=9.6 Hz, 1H),
7.27 (s, 1H), 7.24 (d, J=9.6 Hz, 1H), 6.68 (d, J=9.6 Hz, 1H),
5.74-6.30 (m, 3H), 4.73 (d, J=7.2 Hz, 1H), 4.73 (d, J=7.2 Hz, 1H),
4.40-4.60 (m, 1H), 4.22 (d, J=9.6 Hz, 1H), 3.95 (s, 3H), 3.61 (q,
J=7.2 Hz, 2H), 3.16-3.30 (m, 1H), 2.50-2.77 (m, 2H), 2.20-0.60 (m,
21H), 1.35 (s, 3H) 1.12 (t, J=7.2 Hz, 3H), 1.18 (s, 3H), 1.02 (s,
9H).
Example 40
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-(difluoromethyl)cyclopropyl]-11-ethyl-16-m
ethoxy-3a-methyl-5,8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexa-
decahydro-1
OH-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-
-b]quinoxaline-10-carboxamide
##STR00281##
[0927] Example 40 was prepared in a similar fashion to Example 39,
substituting Intermediate A9 for Intermediate A10 in Step 8.
Example 40 was isolated (70.9 mg) in approximately 92% purity.
Analytic HPLC RetTime: 9.24 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.42H.sub.59F.sub.2N.sub.6O.sub.9S: 861.4. found: 861.4.
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.80 (d, J=9.6 Hz, 1H),
7.25 (s, 1H), 7.23 (d, J=9.6 Hz, 1H), 6.70 (d, J=9.6 Hz, 1H),
5.60-6.10 (m, 3H), 4.69 (d, J=7.2 Hz, 1H), 4.39 (dd, J=12.0, 6.0
Hz, 1H), 4.2 (d, J=9.6 Hz, 1H), 4.03-4.10 (m, 1H), 3.94 (s, 3H),
3.12-3.28 (m, 1H), 2.89-3.05 (m, 1H), 2.50-2.76 (m, 2H), 2.30-0.80
(m, 19H), 1.36 (s, 3H) 1.25 (t, J=7.2 Hz, 3H), 1.10 (s, 3H), 1.04
(s, 9H).
Example 41
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-[(1R,2S)-2-(2,2-difluoroethyl)-1-
-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-1-ethyl-16-methoxy-
-3a-methyl-5,8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahyd-
ro-1
OH-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[1-
1,12-b]quinoxaline-10-carboxamide
##STR00282##
[0929] Example 41 was prepared in a similar fashion to Example 39,
substituting Intermediate A8 for Intermediate A10 in Step 8.
Example 41 was isolated (4.3 mg) in approximately 92% purity.
Analytic HPLC RetTime: 9.36 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.42H.sub.59F.sub.2N.sub.6O.sub.9S: 889.4. found: 889.5.
.sup.1H NMR (300 MHz, CD.sub.3COCD.sub.3) .delta. 7.83 (d, J=7.83
Hz, 1H), 7.19-7.30 (m, 1H), 5.74-6.30 (m, 3H), 4.70 (d, J=7.2 Hz,
1H), 4.19 (dd, J=12.0, 6.0 Hz, 1H), 4.24 (d, J=9.6 Hz, 1H), 4.12
(d, J=12.0, 9.6 Hz, 1H), 3.96 (s, 3H), 3.10-3.26 (m, 1H), 2.56-2.80
(m, 2H), 2.30-0.80 (m, 25H), 1.54 (s, 3H), 1.42 (s, 3H), 1.12 (t,
J=7.2 Hz, 3H), 1.06 (s, 9H).
Example 42 and Example 43
Preparation of
(1aS,5S,8S,9S,10R,22aS)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-1a-ethyl-14-methoxy-9-m-
ethyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,1-
0-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quin-
oxaline-8-carboxamide and
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-1a-ethyl-14-methoxy-9-m-
ethyl-3,6-dioxo-1,1a,3,4,5,6,9,
10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1,1-
0,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00283## ##STR00284## ##STR00285##
[0931] Step 1. Preparation of 43-1: To a solution of Intermediate
mixture D15 (281 mg, 0.81 mmol) and Intermediate 18-2 (290 mg, 0.74
mmol) in MeCN (3.7 mL) was added HATU (308 mg, 0.81 mmol) followed
by DIPEA (640 .mu.L, 3.68 mmol) at rt under an argon atmosphere.
After 17 h, the reaction mixture was concentrated in vacuo, and the
crude residue was purified by silica gel chromatography (0-100%
ethyl acetate/hexanes) to afford 43-1 (121 mg, 1:1 diastereomeric
mixture) as a colorless oil. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.52ClN.sub.4O.sub.7: 687.3. found: 687.5.
[0932] Step 2. Preparation of 43-2: To a solution of diastereomeric
mixture 43-1 (121 mg, 176 .mu.mol), TEA (38 .mu.L, 264 .mu.mol) and
potassium vinyltrifluoroborate (35.4 mg, 264 .mu.mol) in EtOH (0.88
mL) was added PdCl.sub.2(dppf) (14.4 mg, 17.6 .mu.mol). The
reaction mixture was degassed with argon for 10 min and heated to
78.degree. C. After 25 min, the reaction mixture was allowed to
cool to rt and was concentrated in vacuo. The crude residue was
purified by silica gel chromatography (0-100% ethyl
acetate/hexanes) to afford 43-2 (105 mg, 1:1 diastereomeric
mixture) as a yellow oil. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd
for C.sub.38H.sub.55N.sub.4O.sub.7: 679.4. found: 679.5.
[0933] Step 3. Preparation of 43-3: To a solution of diastereomeric
mixture 43-2 (105 mg, 155 .mu.mol) in DCE (31 mL) was added Zhan 1B
catalyst (11.3 mg, 15.5 .mu.mol, Strem) and the reaction mixture
was degassed for 10 minutes with argon. The reaction mixture was
then heated to 100.degree. C. After 15 min, the reaction mixture
was allowed to cool to rt and was concentrated in vacuo. The crude
residue was purified by silica gel chromatography (0-100% ethyl
acetate/hexanes) to afford 43-3 (52.3 mg, 1:1 diastereomeric
mixture) as a light yellow oil. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.36H.sub.51N.sub.4O.sub.7: 651.4. found: 651.5.
[0934] Step 4. Preparation of 43-4: To a solution of diastereomeric
mixture 43-3 (52 mg, 80 .mu.mol) in ethanol (0.4 mL) was added Pd/C
(10 wt % Pd, 9 mg, 8 .mu.mol) at rt under an argon atmosphere. The
atmosphere was replaced with hydrogen and the reaction mixture was
stirred vigorously at rt. After 45 min, the reaction mixture was
diluted with ethyl acetate (1 mL) and was filtered through a pad of
Celite with ethyl acetate washings (3.times.1 mL). The filtrate was
concentrated in vacuo to afford 43-4 (49 mg, 1:1 diastereomeric
mixture), which was used directly in the next step without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.52N.sub.4O.sub.7: 653.4. found: 653.6.
[0935] Step 5. Preparation of 43-5: To a solution of diastereomeric
mixture 43-4 (49 mg, 67 .mu.mol) in DCM (0.5 mL) was added TMSOTf
(60 .mu.L, 0.34 mmol) at rt under an argon atmosphere. After 3 h,
the reaction mixture was added slowly to a 0.25 N aqueous NaOH
solution (precooled to 0.degree. C., 1 mL). The resulting mixture
was diluted with 1 N aqueous HCl solution (5 mL), and was extracted
with DCM (3.times.5 mL). The combined organic extracts were dried
over anhydrous sodium sulfate and were concentrated to afford 43-5
(71 mg, 1:1 diastereomeric mixture) as a tan solid, which was used
directly in the next step without further purification.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.45N.sub.4O.sub.7: 597.3. found: 597.5.
[0936] Step 6. Preparation of Examples 42 and 43: To a solution of
diastereomeric mixture 43-5 (71 mg, .about.67 .mu.mol) and
Intermediate A10 (54 mg, 178 .mu.mol) in MeCN (1.00 mL) was added
HATU (69 mg, 178 .mu.mol) followed by DIPEA (155 .mu.L, 0.89 mmol)
at rt under an argon atmosphere. After 1 h, the reaction mixture
was concentrated in vacuo, and the crude residue was purified by
silica gel chromatography (0-100% ethyl acetate/hexanes). The
fractions containing the desired product were combined and
repurified by preparatory HPLC (Gemini 5u C18 110 .ANG. column,
50-100% MeCN/H.sub.2O, 0.1% trifluoroacetic acid modifier) and were
lyophilized to afford Example 42 (10 mg) and Example 43 (10 mg) as
off white powders. Example 42: Analytic HPLC RetTime: 9.04 min.
[M+H].sup.+ calcd for C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S:
847.4. found: 847.6. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.98
(s, 1H), 7.73 (d, J=8.4 Hz, 2H), 7.20-7.13 (m, 2H), 5.70 (td,
J=55.8, 6.4 Hz, 1H), 5.65 (d, J=3.7 Hz, 1H), 5.44 (br s, 1H),
4.55-4.42 (m, 1H), 4.20-4.03 (m, 1H), 3.87 (s, 3H), 3.17-3.08 (m,
1H), 2.85-2.72 (m, 1H), 2.71-2.59 (m, 1H), 2.18-2.06 (m, 1H),
2.03-1.83 (m, 4H), 1.80-1.53 (m, 5H), 1.50 (br s, 3H), 1.46 (s,
3H), 1.40-1.31 (m, 1H), 1.33-1.09 (m, 5H), 1.06 (s, 9H), 1.05-0.95
(m, 6H), 0.92-0.73 (m, 3H). Example 43: Analytic HPLC RetTime: 9.17
min. [M+H].sup.+ calcd for C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S:
847.4. found: 847.6. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.03
(s, 1H), 7.68 (d, J=9.5 Hz, 1H), 7.14-7.09 (m, 2H), 5.68 (td,
J.sub.H-F=55.5, 6.7 Hz, 1H), 5.59 (d, J=3.7 Hz, 1H), 4.45 (d, J=6.8
Hz, 1H), 4.29 (d, J=12.1 Hz, 1H), 4.12 (s, 1H), 4.08 (dd, J=12.1,
4.3 Hz, 1H), 3.82 (s, 3H), 2.90-2.79 (m, 1H), 2.79-2.70 (m, 1H),
2.66-2.56 (m, 1H), 2.43-2.31 (m, 1H), 1.95-1.85 (m, 2H), 1.75-1.62
(m, 1H), 1.61-1.42 (m, 5H), 1.44 (br s, 3H) 1.40 (s, 3H), 1.34-1.02
(m, 8H), 1.00 (s, 9H), 0.99-0.89 (m, 5H), 0.85-0.74 (m, 3H).
Example 44
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-1a,9-dimethy-
l-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-me-
thanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxal-
ine-8-carboxamide
##STR00286## ##STR00287## ##STR00288##
[0938] Step 1. Preparation of 44-1: HATU (544 mg, 1.43 mmol,
Oakwood) and DIPEA (0.83 mL, 4.76 mmol) were added to a mixture of
18-2 (429 mg, 1.09 mmol) and a Intermediate mixture D6 (395 mg,
1.33 mmol) in 12 mL of acetonitrile under argon. After stirring
overnight, the reaction mixture was concentrated under reduced
pressure and the resulting residue was purified by silica gel
chromatography (0-30% ethyl acetate in hexanes) to produce 44-1
(545 mg; 1:1 mixture of diastereomers) as a white solid. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.35H.sub.50ClN.sub.4O.sub.7:
673.33. found: 673.47.
[0939] Step 2. Preparation of 44-2:
Pd(dppf)Cl.sub.2*CH.sub.2Cl.sub.2 (74 mg, 0.091 mmol, Strem) was
added to a deoxygenated mixture of 44-1 (542 mg, 0.805 mmol),
potassium vinyltrifluoroborate (168 mg, 1.25 mmol), and
triethylamine (0.170 mL, 1.21 mmol) in 9 mL of EtOH at room
temperature. Reaction mixture was heated at 78.degree. C. under
argon for 75 minutes. After cooling to rt, 6 mL of toluene was
added and reaction mixture was concentrated in vacuo. The resulting
residue was purified by silica gel chromatography (0-35% ethyl
acetate in hexanes) to yield 44-2 (438 mg; 1:1 mixture of
diastereomers) as a yellow film. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.37H.sub.53N.sub.4O.sub.7: 665.38. found: 665.55.
[0940] Step 3. Preparation of 44-3 and 44-4: A diastereomeric
mixture 44-2 (437 mg, 0.658 mmol) and Zhan 1B catalyst (81 mg,
0.072 mmol, Strem) in 131 mL of DCE was deoxygenated under argon
for 25 minutes. The mixture was then heated at 95.degree. C. for 50
minutes. An additional 7 mg of Zhan 1B catalyst was added and
reaction mixture was heated at 95.degree. C. for 10 minutes. After
cooling to room temperature, reaction mixture was concentrated in
vacuo. The resulting residue was purified by silica gel
chromatography (0-40% ethyl acetate in hexanes) to yield single
diastereomers 44-3 (143 mg, early eluting component) as a light
yellow film and 44-4 (118 mg, late eluting component) as a light
yellow solid. Early eluting 44-3: LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.35H.sub.49N.sub.4O.sub.7: 637.35. found: 637.45. Late
eluting 44-4: LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.49N.sub.4O.sub.7: 637.35. found: 637.59.
[0941] Step 4. Preparation of 44-5: Palladium on carbon (10 wt %
Pd, 48 mg, 0.045 mmol) was added to a solution of 44-3 (143 mg,
0.225 mmol) in 6 mL of ethanol. The atmosphere was replaced with
hydrogen and the reaction stirred for 90 minutes. The reaction
mixture was filtered over Celite and washed with ethyl acetate.
Filtrate was concentrated in vacuo to yield 44-5 (130 mg) as brown
solid film, which was used in the next step without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.51N.sub.4O.sub.7: 639.37. found: 639.53.
[0942] Step 5. Preparation of 44-6: TMSOTf (0.53 mL, 2.91 mmol) was
added dropwise to a solution of 44-5 (130 mg, 1.27 mmol) in 3.8 mL
of dichloromethane under argon at room temperature. After one hour,
an additional 0.22 mL of TMSOTf was added. After an additional
hour, 0.20 mL of TMSOTf was added. After 40 minutes, 0.25 mL of
TMSOTf was added. After one hour, reaction mixture was taken up in
10 mL of dichloromethane and quenched by addition of 20 mL of 1 N
aqueous HCl with stirring. Layers were separated and aqueous was
extracted with dichloromethane (3.times.30 mL). Combined organics
were washed with brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo to yield 44-6 (113 mg) as an
off white solid, which was used in the next step without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.31H.sub.43N.sub.4O.sub.7: 583.31. found: 583.45.
[0943] Step 6. Preparation of Example 44: HATU (53 mg, 0.139 mmol)
and DIPEA (0.080 mL, 0.459 mmol) were added to a mixture of 44-6
(51 mg, 0.088 mmol) and Intermediate A10 (49 mg, 0.161 mmol) in 1.5
mL of MeCN under argon. After stirring for overnight, an additional
13 mg of Intermediate A10 was added. After one hour, reaction
mixture was taken up in 15 mL of ethyl acetate and poured into 20
mL of 1 N aqueous HCl. Layers were separated and aqueous was
extracted three times with ethyl acetate. Combined organics were
washed with water and brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The resulting residue was
purified by silica gel chromatography (0-40% ethyl acetate in
hexanes) to yield Example 44 (41 mg) as an off white solid.
Analytic HPLC RetTime: 8.86 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.40H.sub.55F.sub.2N.sub.6O.sub.9S: 833.36. found:
833.51. .sup.1H NMR (400 MHz, CD.sub.3OD): 7.79 (d, J=10 Hz, 1H),
7.28-7.21 (m, 2H), 6.77 (d, J=8.4 Hz, 1H), 5.81 (td, J.sub.H-F=56
Hz, J=6.4 Hz, 1H), 5.73-5.70 (m, 1H), 4.56 (d, J=7.2 Hz, 1H), 4.40
(d, J=11.6 Hz, 1H), 4.26-4.16 (m, 2H), 3.93 (s, 3H), 3.05-2.91 (m,
1H), 2.90-2.82 (m, 1H), 2.77-2.68 (m, 1H), 2.06-1.94 (m, 2H),
1.88-1.74 (m, 1H), 1.72-1.58 (m, 3H), 1.58-1.44 (m, 4H), 1.53 (s,
3H), 1.51 (s, 3H), 1.43-1.36 (m, 1H), 1.12-1.02 (m, 2H), 1.09 (s,
9H), 1.07 (d, J=4 Hz, 3H), 1.00-0.94 (m, 2H), 0.92-0.84 (m, 3H),
0.16-0.11 (m, 1H).
Example 45
Preparation of
(1aS,5S,8S,9S,10R,22aS)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-methoxy-1a,9-dimethy-
l-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-me-
thanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxal-
ine-8-carboxamide
##STR00289##
[0945] Example 45 was prepared in a similar fashion to Example 44,
substituting late eluting 44-4 for early eluting 44-3 in step 4.
Example 45 was isolated (23 mg) as an off white solid. Analytic
HPLC RetTime: 8.92 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.55F.sub.2N.sub.6O.sub.9S: 833.36. found: 833.54.
.sup.1H NMR (400 MHz, CD.sub.3OD): 7.79 (d, J=9.2 Hz, 1H),
7.25-7.19 (m, 2H), 6.55 (d, J=5.2 Hz, 1H), 5.78 (td, J.sub.H-F=61
Hz, J=6 Hz, 1H), 5.52-5.48 (m, 1H), 4.58 (d, J=6.4 Hz, 1H), 4.52
(d, J=12 Hz, 1H), 4.17-4.10 (m, 1H), 4.04 (d, J=6.4 Hz, 1H), 3.93
(s, 3H), 3.22-3.14 (m, 1H), 2.88-2.80 (m, 1H), 2.78-2.66 (m, 1H),
2.08-1.90 (m, 2H), 1.76-1.64 (m, 1H), 1.63-1.50 (m, 7H), 1.51 (s,
3H), 1.47-1.36 (m, 2H), 1.46 (s, 3H), 1.18-1.06 (m, 1H), 1.12 (s,
9H), 1.07 (m, 3H), 1.00-0.80 (m, 4H), 0.10-0.04 (m, 1H).
Example 46 and 47
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-18,18-difluoro-14-metho-
xy-9-methyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro--
8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12--
b]quinoxaline-8-carboxamide and
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-18-fluoro-14-methoxy-9--
methyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,-
10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]qui-
noxaline-8-carboxamide
##STR00290## ##STR00291## ##STR00292## ##STR00293##
[0947] Step 1. Preparation of 46-1: A mixture of Intermediate B1
(627 mg, 2.08 mmol), Intermediate E3 (548 mg, 1.91 mmol) and cesium
carbonate (744 mg, 2.28 mmol) in 7 mL of DMF was stirred at
85.degree. C. under argon for 36 hours. Reaction mixture was cooled
to room temperature and poured into 30 mL of water and aqueous was
extracted with ethyl acetate (3.times.30 mL). Combined organics
were washed with water, brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. The
resulting residue was purified by silica gel chromatography (0-30%
ethyl acetate in hexanes) to yield 46-1 (891 mg) as a white solid.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.27H.sub.36F.sub.2N.sub.3O.sub.6: 536.25. found: 536.35.
[0948] Step 2. Preparation of 46-2: Quinoxaline ether 46-1 (478 mg,
0.893 mmol) was dissolved in 4.2 mL of tert-butyl acetate and 1.1
mL of dichloromethane at room temperature. MeSO.sub.3H (0.30 mL,
4.67 mmol) was added dropwise and reaction mixture stirred at rt
for 2 h. The reaction mixture was transferred to a stirred mixture
of EtOAc (20 mL) and saturated aqueous NaHCO.sub.3 (30 mL). The
phases were separated, and the aqueous phase was extracted with
EtOAc (2.times.20 mL). The combined organic phase was dried over
anhydrous Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to
afford amine 46-2 as a yellow solid film (346 mg). LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.22H.sub.28F.sub.2N.sub.3O.sub.4:
436.20. found: 436.29.
[0949] Step 3. Preparation of 46-3: HATU (396 mg, 1.04 mmol,
Oakwood) and DIPEA (0.57 mL, 3.29 mmol) were added to a mixture of
46-2 (345 mg, 0.793 mmol) and Intermediate D11 (260 mg, 0.965 mmol)
in 9 mL of acetonitrile under argon. After stirring overnight, the
reaction mixture was concentrated under reduced pressure and the
resulting residue was purified by silica gel chromatography (0-40%
ethyl acetate in hexanes) to yield 46-3 (545 mg) as a clear solid
film. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49F.sub.2N.sub.4O.sub.7: 687.35. found: 687.57.
[0950] Step 4. Preparation of 46-4: A mixture of 46-3 (480 mg,
0.699 mmol) and Zhan 1B catalyst (61 mg, 0.083 mmol, Strem) in 140
mL of DCE was deoxygenated with argon for 18 minutes. The mixture
was then heated at 95.degree. C. for 70 minutes. An additional 20
mg of Zhan 1B catalyst was added and mixture stirred at 95.degree.
C. for one hour. After cooling to room temperature, reaction
mixture was concentrated in vacuo. The resulting residue was
purified by silica gel chromatography (0-35% ethyl acetate in
hexanes) to yield an inseparable mixture of 46-4 (major), and
approximately 15% of 47-1 (minor; 233 mg total) as an off white
solid. Major component 46-4: LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.45F.sub.2N.sub.4O.sub.7: 665.38. found: 665.50. Minor
component 47-1: LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.44FN.sub.4O.sub.7: 639.31. found: 639.49.
[0951] Step 5. Preparation of mixture of 46-5 and 47-2: Palladium
on carbon (10 wt % Pd, 70 mg, 0.066 mmol) was added to a solution
of the mixture of 46-4 and 47-1 (232 mg, 0.353 mmol) from the
previous step in 9 mL of ethanol. The atmosphere was replaced with
hydrogen and stirred for 7 h. The reaction was filtered over
Celite, washing with ethanol. Filtrate was concentrated in vacuo to
yield a mixture of 46-5 (major) and 47-2 (minor; 216 mg total) as
an off white solid, which was used in the next step without further
purification. Major component 46-5: LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.34H.sub.47F.sub.2N.sub.4O.sub.7: 661.33. found:
661.52. Minor component 47-2: LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.34H.sub.48FN.sub.4O.sub.7: 643.34. found:
643.57.
[0952] Step 6. Preparation of mixture of 46-6 and 47-3: TMSOTf
(0.35 mL, 1.90 mmol) was added dropwise to a solution of a mixture
of 46-5 and 47-2 (215 mg, 0.326 mmol) from the previous step in 6.5
mL of dichloromethane under argon at rt. After 1 h, an additional
0.18 mL of TMSOTf was added. After an additional hour, 0.30 mL of
TMSOTf was added. After 2 h, 0.18 mL of TMSOTf was added. After 1
h, an additional 0.18 mL of TMSOTf was added. After 45 minutes,
reaction mixture was taken up in 25 mL of dichloromethane and
quenched by addition of 30 mL of 1 N aqueous HCl with stirring. The
aqueous layer was extracted with dichloromethane (3.times.40 mL).
Combined organics were washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to yield an
inseparable mixture of 46-6 (major) and 47-3 (minor; 187 mg total),
which was used in the next step without further purification. Major
component 46-6: LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.30H.sub.39F.sub.2N.sub.4O.sub.7: 605.27. found: 605.44.
[0953] Minor component 47-3: LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.30H.sub.39FN.sub.4O.sub.7: 587.28. found:
587.38.
[0954] Step 7. Preparation of Example 46 and Example 47: HATU (160
mg, 0.421 mmol, Oakwood) and DIPEA (0.25 mL, 1.44 mmol) were added
to a mixture of 46-6 and 47-3 (150 mg, 0.248 mmol) from the
previous step and Intermediate A10 (150 mg, 0.496 mmol) in 6.5 mL
of acetonitrile under argon. After stirring for overnight, reaction
mixture was taken up in 30 mL of ethyl acetate and poured into 30
mL of 1 N aqueous HCl. The aqueous layer was extracted three times
with ethyl acetate. Combined organics were washed with water and
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The resulting residue was purified by silica
gel chromatography (0-50% ethyl acetate in hexanes) and reverse
phase prep HPLC (50-100% acetonitrile in water, with 0.1%
trifluoroacetic acid buffer) to yield the trifluoroacetic acid salt
of Example 46 (107 mg) as a light yellow solid and the
trifluoroacetic acid salt of the 1:1 mixture of diastereomers of
Example 47 (12 mg) as a light yellow solid. Example 46: Analytic
HPLC RetTime: 8.60 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.51F.sub.4N.sub.6O.sub.9S: 855.33. found: 855.63.
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 9.23 (s, 1H), 7.94 (d,
J=9.2 Hz, 1H), 7.31 (dd, J=9.2, 2.8 Hz, 1H), 7.28 (d, J=2.8 Hz,
1H), 5.78 (td, J.sub.H-F=66 Hz, J=6.8 Hz, 1H), 5.68-5.66 (m, 1H),
4.57 (d, J=6.4 Hz, 1H), 4.41 (d, J=12 Hz, 1H), 4.35 (s, 1H),
4.22-4.16 (dd, J=12, 4 Hz, 1H), 3.97 (s, 3H), 3.72-3.66 (m, 1H),
2.86-2.76 (m, 1H), 2.64-2.48 (m, 1H), 2.11-1.94 (m, 3H), 1.86-1.74
(m, 3H), 1.73-1.62 (m, 1H), 1.58-1.54 (m, 2H), 1.50 (s, 3H),
1.49-1.44 (m, 1H), 1.42-1.38 (m, 1H), 1.11-1.04 (m, 4H), 1.09 (s,
9H), 1.02-0.94 (m, 2H), 0.93-0.86 (m, 2H), 0.78-0.66 (m, 1H),
0.54-0.46 (m, 1H). Example 47 (1:1 mixture of diastereomers):
Analytic HPLC RetTime: 8.45 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.39H.sub.52F.sub.3N.sub.6O.sub.9S: 837.34. found: 837.63.
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 9.13 (s, 1H), 7.89 (d,
J=8.8 Hz, 1H), 7.27 (dd, J=9.2, 2.8 Hz, 1H), 7.24 (d, J=2.8 Hz,
1H), 5.99-5.43 (m, 1H), 5.79 (td, J.sub.H-F=55 Hz, J=6.8 Hz, 1H),
5.53-5.50 (m, 1H), 4.57-4.44 (m, 2H), 4.11 (s, 1H), 4.35 (s, 1H),
4.22-4.13 (dd, J=12.4, 4 Hz, 1H), 3.95 (s, 3H), 3.83-3.79 (m, 1H),
2.94-2.80 (m, 2H), 2.28-2.14 (m, 1H), 2.06-1.96 (m, 2H), 1.88-1.69
(m, 4H), 1.58-1.54 (m, 2H), 1.51 (s, 3H), 1.44-1.36 (m, 1H),
1.32-1.26 (m, 1H), 1.14-1.04 (m, 4H), 1.10 (s, 9H), 1.02-0.86 (m,
4H), 0.74-0.64 (m, 1H), 0.58-0.48 (m, 1H).
Example 48
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-{(1R,2S)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-ethenylcyclopropyl}-14-methoxy-9-methyl-3,6-dioxo-1,1a,3,4,5,6-
,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1-
,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00294##
[0956] Step 1: Preparation of Example 48: To a suspension of acid
18-7 (9.7 mg, 0.017 mmol) and Intermediate A1 (13 mg, 0.049 mmol)
in MeCN (0.4 mL) was added DIPEA (40 .mu.L, 0.23 mmol). To the
resulting solution was added HATU (12.5 mg, 0.033 mmol). The
reaction was stirred at rt for 1 h and was diluted with EtOAc (2
mL), 0.2 M aqueous HCl (1 mL), and brine (1 mL). The phases were
separated and the aqueous phase was extracted with EtOAc (3.times.2
mL). The combined organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue that was dissolved in CH.sub.2Cl.sub.2 and adsorbed onto 1
g silica gel. Purification by silica gel chromatography (10% to 50%
acetone in hexanes) provided a residue that was lyophilized from
water and MeCN to provide Example 48 as a white amorphous solid
(8.4 mg). Analytic HPLC RetTime: 8.52 min. LCMS-ESI.sup.+ (m/z):
[M+H]*calcd for C.sub.39H.sub.53N.sub.6O.sub.9S: 781.4. found:
781.2. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.91 (s, 1H), 7.83
(d, J=9.1 Hz, 1H), 7.19 (dd, J=9.1, 2.7 Hz, 1H), 7.10 (d, J=2.7 Hz,
1H), 6.73 (s, 1H), 5.86-5.72 (m, 1H), 5.57 (d, J=3.8 Hz, 1H), 5.48
(d, J=9.9 Hz, 1H), 5.27-5.15 (m, 1H), 5.15-5.07 (m, 1H), 4.48-4.35
(m, 3H), 4.12 (dd, J=11.8, 4.1 Hz, 1H), 3.94 (s, 3H), 3.81-3.71 (m,
1H), 2.98-2.75 (m, 4H), 2.16-2.09 (m, 1H), 1.94 (dd, J=8.2, 5.8 Hz,
1H), 1.87-1.24 (m, 9H), 1.17 (d, J=7.4 Hz, 3H), 1.09 (s, 9H),
1.04-0.91 (m, 5H), 0.75-0.65 (m, 1H), 0.52-0.42 (m, J=6.0 Hz,
1H).
Example 49
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-2-(difluorometh-
yl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-15-methoxy-10-
-methyl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadecahyd-
ro-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6]dio-
xadiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00295##
[0958] Step 1: Preparation of Example 49: To a suspension of acid
33-4 (30 mg, 0.049 mmol) and Intermediate A10 (31 mg, 0.10 mmol) in
MeCN (700 .mu.L) was added DIPEA (70 .mu.L, 0.40 mmol). HATU (32
mg, 0.084 mmol) was added to the resulting solution, and the
reaction was stirred at rt for 1.5 h. An additional portion of
Intermediate A10 (6 mg, 0.02 mmol) was then added. The reaction was
stirred an additional 30 min and was then diluted with EtOAc (30
mL), 0.2 M aqueous HCl (15 mL) and brine (15 mL). The phases were
separated and the aqueous phase was extracted with EtOAc (30 mL).
The combined organic phase was dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue. This residue was dissolved in CH.sub.2Cl.sub.2 and
adsorbed onto 2 g silica gel. Purification by silica gel
chromatography (10% to 50% acetone in hexanes) provided an
amorphous residue that was lyophilized from water and MeCN to
provide Example 49 as a white amorphous solid (30.5 mg). Analytic
HPLC RetTime: 9.15 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.57F.sub.2N.sub.6O.sub.9S: 859.4. found: 859.2.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.86 (s, 1H), 7.82 (d,
J=9.1 Hz, 1H), 7.45 (s, 1H), 7.18 (dd, J=9.1, 2.7 Hz, 1H), 7.08 (d,
J=2.7 Hz, 1H), 6.14-5.71 (m, 1H), 5.61 (d, J=3.7 Hz, 1H), 5.28 (d,
J=9.8 Hz, 1H), 5.00 (d, J=7.4 Hz, 1H), 4.49 (d, J=7.0 Hz, 1H),
4.42-4.31 (m, 2H), 4.12 (dd, J=11.6, 4.0 Hz, 1H), 3.93 (s, 3H),
3.00-2.63 (m, 4H), 2.25-2.16 (m, 1H), 2.09-1.90 (m, 4H), 1.81-0.95
(m, 26H), 0.92-0.75 (m, 3H), 0.57-0.45 (m, 1H), 0.44-0.36 (m,
1H).
Example 50
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-9-(cyanomethyl)-N-[(1R,2R)-2-(difluo-
romethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-meth-
oxy-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10--
methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinox-
aline-8-carboxamide
##STR00296##
[0960] Example 50 was prepared in a similar fashion to Example 1,
substituting Intermediate B8 for Intermediate B4 in step 1. Example
50 was purified by reverse phase HPLC (Gemini column, 58-98%
ACN/H.sub.2O+0.1% TFA) and lyophilized to afford solid (5 mg) as a
TFA salt. Analytic HPLC RetTime: 8.29 min. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.40H.sub.51F.sub.2N.sub.7O.sub.9S:
844.94. found: 844.58. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
9.71 (s, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.22 (m, 2H), 6.25 (m, 1H),
6.08-5.80 (m, 1H), 4.39 (m, 1H), 4.29 (m, 2H), 4.13 (m, 1H), 3.92
(s, 3H), 3.65 (m, 1H), 3.06-2.83 (m, 4H), 2.55 (m, 1H), 2.14-1.47
(m, 17H), 1.03 (s, 9H), 0.92 (m, 4H), 0.65 (m, 1H), 0.45-0.43 (m,
1H).
Example 51
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-N-{(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-ethylcyclopropyl}-1-ethyl-16-methoxy-3a-methyl-5,8-dioxo-1,2-
,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-1
OH-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-
-b]quinoxaline-10-carboxamide
##STR00297##
[0962] Example 51 was prepared in a similar fashion to Example 39,
substituting Intermediate A3 for Intermediate A10 in step 8.
Example 51 was isolated (12.3 mg) in approximately 96.5% purity.
Analytic HPLC RetTime: 9.38 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.43H.sub.63N.sub.6O.sub.9S: 839.4. found: 839.5.
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.60 (d, J=8.4 Hz, 1H),
6.98-7.08 (m, 2H), 6.53 (d, J=9.6 Hz, 1H), 5.57-5.83 (m, 2H), 4.52
(d, J=8.4 Hz, 2H), 4.24 (dd, J=10.8, 6.0 Hz, 1H), 4.02 (d, J=9.6
Hz, 1H), 3.82 (dd, J=10.8, 2.4 Hz, 1H), 3.73 (s, 3H), 2.93-3.10 (m,
1H), 2.80-2.90 (m, 2H), 2.30-2.58 (m, 2H), 0.60-2.10 (m, 32H), 0.84
(s, 9H).
Example 52
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-11-ethyl-N-[(1R,2S)-2-(2-fluoroeth-
yl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-16-methoxy-3a-
-methyl-5,8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro--
10H-9,12-methanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-
-b]quinoxaline-10-carboxamide
##STR00298##
[0964] Example 52 was prepared in a similar fashion to Example 39,
substituting Intermediate A6 for Intermediate A10 in step 8.
Example 52 was isolated (12.3 mg) in approximately 96.5% purity.
Analytic HPLC RetTime: 8.60 min. Analytic HPLC RetTime: 9.31 min.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.44H.sub.64FN.sub.6O.sub.9S: 871.4. found: 871.5. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 7.81 (d, J=8.4 Hz, 1H), 7.20-7.30 (m,
2H), 6.73 (d, J=9.6 Hz, 1H), 5.75-6.02 (m, 2H), 4.74 (d, J=8.4 Hz,
2H), 4.54 (t, J=6.0 Hz, 2H), 4.36-4.49 (m, 1H), 4.23 (d, J=9.6 Hz,
1H), 4.04 (dd, J=12.0, 2.4 Hz, 1H), 3.75 (s, 3H), 3.28-3.16 (m,
1H), 2.50-2.70 (m, 2H), 2.30-0.80 (m, 35H), 1.04 (s, 9H).
Example 53
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-(difluoromethyl)cyclopropyl]-9-ethyl-18,18-difluoro-3,6-dioxo--
1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopr-
opa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carbox-
amide
##STR00299##
[0966] Example 53 was prepared similarly to Example 17 substituting
Intermediate E4 for Intermediate E3 in Step 1 and Intermediate A9
for Intermediate A10 in Step 7. Example 53 was isolated (8.8 mg) as
a white solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.48F.sub.4N.sub.6O.sub.8S: 825.32. found: 825.75.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.13 (s, 1H), 8.15 (d,
J=8.2 Hz, 1H), 7.91-7.74 (m, 2H), 7.69 (t, J=7.6 Hz, 1H), 6.92 (s,
1H), 5.47 (d, J=9.6 Hz, 1H), 4.48 (t, J=10.3 Hz, 2H), 4.36 (d,
J=9.4 Hz, 1H), 4.12 (dd, J=12.1, 3.6 Hz, 1H), 3.70-3.59 (m, 1H),
3.08-2.75 (m, 1H), 2.58-2.38 (m, 1H), 2.14 (t, J=6.8 Hz, 1H),
1.95-1.67 (m, 4H), 1.47 (tt, J=13.9, 7.1 Hz, 4H), 1.35 (s, 2H),
1.20 (t, J=7.3 Hz, 3H), 1.15-0.64 (m, 19H), 0.51 (q, J=6.4 Hz,
1H).
Example 54
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-{(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-ethylcyclopropyl}-9-ethyl-18,18-difluoro-3,6-dioxo-1,1a,3,4,5,-
6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][-
1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00300##
[0968] Example 54 was prepared similarly to Example 53 replacing
Intermediate A9 with Intermediate A3. Example 54 was isolated (10.0
mg) as a white solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.52F.sub.2N.sub.6O.sub.8S: 803.35. found: 803.79.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.88 (s, 1H), 8.12 (d,
J=8.2 Hz, 1H), 7.88-7.69 (m, 2H), 7.66 (t, J=7.6 Hz, 1H), 6.68 (s,
1H), 5.95 (d, J=3.4 Hz, 1H), 5.46 (d, J=9.4 Hz, 1H), 4.45 (dd,
J=13.8, 9.7 Hz, 2H), 4.09 (dd, J=12.0, 3.6 Hz, 2H), 3.71-3.57 (m,
1H), 2.53 (dd, J=21.4, 14.6 Hz, 1H), 1.85-1.39 (m, 10H), 1.38-0.96
(m, 20H), 1.01 (dd, J=17.2, 9.5 Hz, 3H), 1.04-0.78 (m, 6H), 0.70
(s, 1H), 0.49 (dd, J=12.7, 6.3 Hz, 1H).
Example 55
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-3,6-dioxo-14-(2-
,2,2-trifluoroethoxy)-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro--
8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12--
b]quinoxaline-8-carboxamide
##STR00301## ##STR00302##
[0970] Intermediate 55-1 was prepared by following Steps 1 through
6 of Example 1, substituting for Intermediate E2 for Intermediate
E1 in Step 1. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.49N.sub.4O.sub.7: 625.36. found: 625.25.
[0971] Step 1. Preparation of 55-2. Quinoxalinol 55-1 (24 mg, 0.038
mmol) was suspended in DMF (2 mL) and treated with Cs.sub.2CO.sub.3
(63 mg, 0.19 mmol) and 2,2,2-trifluoroethyl
trifluoromethanesulfonate (0.055 mL, 0.38 mmol). The reaction
mixture was stirred at RT for 5 h, then diluted with EtOAc. The
organic layer was washed with H.sub.2O and brine, dried over
MgSO.sub.4, filtered and concentrated under reduced pressure to
afford 55-2, which was carried on without further purification.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.50F.sub.3N.sub.4O.sub.7: 707.36. found: 707.38.
[0972] Step 2. Preparation of 55-3. Trifluoroethyl ether 55-2
(0.038 mmol theoretical) was treated with DCM (4 mL) and TMSOTf
(0.14 mL, 0.77 mmol) at RT. After 1 h, the reaction was quenched by
addition of 1M NaOH (2 mL). After stirring vigorously for 5 min,
the mixture was poured into a separatory funnel followed by 10% HCl
(20 mL). The aqueous layer was extracted 3.times. with DCM. The
combined organics were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure to afford 55-3, which was
carried on without further purification. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.32H.sub.42F.sub.3N.sub.4O.sub.7:
651.30. found: 651.18.
[0973] Step 3. Preparation of Example 55. Carboxcylic acid 55-3
(0.038 mmol theoretical) was treated with intermediate A10 (23 mg,
0.077 mmol), TBTU (25 mg, 0.077 mmol), DMAP (9 mg, 0.077 mmol), DCM
(1 mL) and DIPEA (0.134 mL, 0.768 mmol). The reaction mixture was
stirred at RT for 20 h, then concentrated under reduced pressure
and purified by reverse phase HPLC to afford Example 55 as a TFA
salt (7 mg, 18% over 3 steps). LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.41H.sub.54F.sub.5N.sub.6O.sub.9S: 901.36. found: 902.08.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.18 (s, 1H), 7.86 (d,
J=9.1 Hz, 1H), 7.32 (dd, J=9.1, 2.8 Hz, 1H), 7.25 (d, J=2.7 Hz,
1H), 6.02-5.63 (m, 2H), 4.76-4.62 (m, 2H), 4.56 (d, J=7.1 Hz, 1H),
4.39 (t, J=6.0 Hz, 2H), 4.15 (dt, J=17.2, 8.6 Hz, 1H), 3.74 (dd,
J=6.7, 2.8 Hz, 1H), 3.05-2.89 (m, 1H), 2.82 (td, J=13.2, 4.2 Hz,
1H), 2.65-2.50 (m, 1H), 2.02 (d, J=10.4 Hz, 2H), 1.78 (dt, J=23.5,
10.7 Hz, 3H), 1.68-1.26 (m, 14H), 1.22 (t, J=7.3 Hz, 3H), 1.10 (s,
9H), 0.97 (d, J=2.5 Hz, 2H), 0.95-0.84 (m, 2H), 0.71 (s, 1H), 0.51
(t, J=9.8 Hz, 1H).
Example 56
Preparation of
(3aR,7S,10S,11S,12R,24aR)-7-tert-butyl-11-ethyl-N-[(1R,2R)-2-ethyl-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-16-methoxy-3a-methyl-5,-
8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-10H-9,12-m-
ethanocyclopenta[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxa-
line-10-carboxamide
##STR00303##
[0975] Example 56 was prepared in a similar fashion to Example 39,
substituting Intermediate A9 for Intermediate A3 in Step 8. Example
56 was isolated (8.8 mg, 0.0103 mmol, 53.7%). Analytic HPLC
RetTime: 9.56 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.44H.sub.65N.sub.6O.sub.9S: 853.45. found: 853.5. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 7.81 (d, J=9.6 Hz, 1H), 7.20-7.30 (m,
2H), 6.73 (d, J=9.6 Hz, 1H), 5.76-6.01 (m, 2H), 4.75 (d, J=8.4 Hz,
1H), 4.46 (dd, J=12.0, 6.0 Hz, 1H), 4.23 (d, J=9.6 Hz, 1H),
4.00-4.08 (m, 1H), 3.95 (s, 3H), 2.50-2.78 (m, 3H), 0.80-2.30 (m,
30H), 1.54 (s, 3H), 1.35 (s, 3H), 1.05 (s, 9H).
Example 57
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2S)-1-[(cyclopropyl-
sulfonyl)carbamoyl]-2-(2,2-difluoroethyl)cyclopropyl]-15-methoxy-10-methyl-
-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadecahydro-1H,9-
H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6]dioxadiaza-
cyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00304##
[0977] Step 1. Preparation of Example 57. To a suspension of acid
33-4 (14.9 mg, 0.0245 mmol) and amine hydrochloride A-7 (16.3 mg,
0.0535 mmol) in MeCN (500 .mu.L) was added DIPEA (40 .mu.L, 0.23
mmol). HATU (15.5 mg, 0.0408 mmol) was added to the resulting
solution, and the reaction was stirred at rt for 17 h. The reaction
was then diluted with EtOAc (2 mL), 0.2 M aqueous HCl (1.5 mL) and
brine (1.5 mL). The phases were separated and the aqueous phase was
extracted with EtOAc (4.times.1.5 mL). The combined organic phase
was dried over Na.sub.2SO.sub.4, filtered, and concentrated to
afford a crude residue. This residue was dissolved in
CH.sub.2Cl.sub.2 and was concentrated onto 1.5 g silica gel.
Purification by silica gel chromatography (10% to 40% acetone in
hexanes) provided an amorphous residue that was lyophilized from
water and MeCN to provide Example 57. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.42H.sub.57F.sub.2N.sub.6O.sub.9S:
859.4. found: 859.0. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
10.00 (s, 1H), 7.82 (d, J=9.1 Hz, 1H), 7.19 (dd, J=9.1, 2.7 Hz,
1H), 7.09 (d, J=2.6 Hz, 1H), 6.75 (s, 1H), 6.07-5.57 (m, 2H), 5.26
(d, J=9.8 Hz, 1H), 5.01 (d, J=7.4 Hz, 1H), 4.50-4.29 (m, 3H), 4.12
(dd, J=11.7, 3.9 Hz, 1H), 3.93 (s, 3H), 3.00-2.62 (m, 4H),
2.34-0.96 (m, 33H), 0.95-0.78 (m, 1H), 0.51 (dd, J=13.0, 7.9 Hz,
1H), 0.39 (d, J=4.2 Hz, 1H).
Example 58
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2S)-2-(2,2-difluoro-
ethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-15-methoxy-
-10-methyl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadeca-
hydro-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6]-
dioxadiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00305##
[0979] Step 1. Preparation of Example 58. To a suspension of acid
33-4 (14.5 mg, 0.0238 mmol) and amine hydrochloride A-8 (16.0 mg,
0.0502 mmol) in MeCN (500 .mu.L) was added DIPEA (40 .mu.L, 0.23
mmol). HATU (15.5 mg, 0.0408 mmol) was added to the resulting
solution, and the reaction was stirred at rt for 17 h. The reaction
was then diluted with EtOAc (2 mL), 0.2 M aqueous HCl (1.5 mL) and
brine (1.5 mL). The phases were separated and the aqueous phase was
extracted with EtOAc (4.times.1.5 mL). The combined organic phase
was dried over Na.sub.2SO.sub.4, filtered, and concentrated to
afford a crude residue. This residue was dissolved in
CH.sub.2Cl.sub.2 and was concentrated onto 1.5 g silica gel.
Purification by silica gel chromatography (10% to 40% acetone in
hexanes) provided an amorphous residue that was lyophilized from
water and MeCN to provide Example 58. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.43H.sub.59F.sub.2N.sub.6O.sub.9S: 873.4. found:
873.3. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.72 (s, 1H), 7.82
(d, J=9.1 Hz, 1H), 7.19 (dd, J=9.1, 2.7 Hz, 1H), 7.09 (d, J=2.7 Hz,
1H), 6.82 (s, 1H), 6.12-5.54 (m, 2H), 5.25 (d, J=9.8 Hz, 1H), 5.01
(d, J=7.2 Hz, 1H), 4.50-4.30 (m, 3H), 4.13 (dd, J=11.7, 4.2 Hz,
1H), 3.93 (s, 3H), 3.03-2.65 (m, 4H), 2.34-0.97 (m, 33H), 0.94-0.76
(m, 3H), 0.60-0.45 (m, 1H), 0.45-0.34 (m, 1H).
Example 59
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-1-[(cyclopropyl-
sulfonyl)carbamoyl]-2-(difluoromethyl)cyclopropyl]-10-ethyl-19,19-difluoro-
-15-methoxy-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadec-
ahydro-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6-
]dioxadiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00306## ##STR00307##
[0981] Step 1. Preparation of 59-1. A solution of Intermediate D16
(0.50 g, 1.6 mmol) in DMF (7 mL) was treated subsequently with COMU
(0.80 g, 1.9 mmol), DIPEA (1.2 mL, 6.7 mmol) and Intermediate 17-2
(0.65 g, 1.3 mmol) and stirred overnight at rt. The reaction was
quenched with 1 M citric acid solution (5 mL) and extracted with
EA. The combined organics were washed with brine, dried over
anhydrous MgSO.sub.4 and concentrated in vacuo. The resulting
residue was purified by silica gel chromatography (15-100% EA/hex)
to afford 59-1. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.55F.sub.2N.sub.4O.sub.7: 741.88. found: 741.51.
[0982] Step 2. Preparation of 59-2. A solution of 59-1 (0.51 g,
0.69 mmol) in DCE (140 mL) is sparged with argon for 30 min prior
to addition of Zhan 1B catalyst (0.051 g, 0.07 mmol). The reaction
was heated to 85.degree. C. for 45 min, and another portion of Zhan
1B catalyst was added. After an additional 30 min, the reaction was
cooled to rt, concentrated in vacuo and purified by silica gel
chromatography (5-100% EA/hex) to produce 59-2. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.38H.sub.51F.sub.2N.sub.4O.sub.7:
713.83. found: 713.54.
[0983] Step 3. Preparation of 59-3. A solution of 59-2 was taken up
in EtOH (8 mL). Pd/C (0.072 g, 10% w/w) was added and the
atmosphere replaced with H.sub.2. After 1 h, additional catalyst
was added. After 4 h, EA and additional catalyst was added. After
an additional 3 h, the reaction was filtered, concentrated in
vacuo, and the residue taken up in EtOH (8 mL) and treated with 0.5
g Pd/C (10% w/w) and the atmosphere replaced with H.sub.2. The
reaction was stirred overnight, and then worked up again as
previously described to produce of 59-3 that was used subsequently
without further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.53F.sub.2N.sub.4O.sub.7: 715.85. found: 715.52.
[0984] Step 4. Preparation of 59-4. A solution of 59-3 (0.40 g,
0.56 mmol) in DCM (1.5 mL) was treated with 2.5 mL TFA at rt. After
1.5 h, the reaction was concentrated in vacuo. The residue was
taken up in EA, washed with saturated aqueous NaHCO.sub.3, brine
and then dried over anhydrous MgSO.sub.4. Concentration in vacuo
produced 59-4 that was used subsequently without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.45F.sub.2N.sub.4O.sub.7: 659.74. found: 659.56.
[0985] Step 5. Preparation of Example 59: A solution of 59-4 (0.20
g, 0.30 mmol) in DMF (2 mL) was treated subsequently with HATU
(0.21 g, 0.55 mmol), DIPEA (0.27 mL, 1.5 mmol), DMAP (0.056 g, 0.46
mmol), and Intermediate A9 (0.13 g, 0.46 mmol) and stirred for 5 h
at rt. The reaction mixture is purified by preparatory HPLC to
produce the TFA salt of Example 59. Analytic HPLC RetTime: 9.20
min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.55F.sub.4N.sub.6O.sub.9S: 895.98. found: 895.60.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.31 (s, 1H); 7.94 (d,
J=9.2 Hz, 1H); 7.32 (dd, J=9.2, 2.4 Hz, 1H); 7.21 (d, J=2.4 Hz,
1H); 5.98 (br s, 1H); 5.85 (td, J.sub.H-F=55.2 Hz, J=6 Hz, 1H);
4.94 (d, J=7.6 Hz, 1H); 4.58 (d, J=7.2 Hz, 1H); 4.35 (d, J=7.2 Hz,
1H); 4.33 (br s, 1H); 4.18 (dd, J=12, 3.6 Hz, 1H); 3.97 (brs, 3H);
2.98 (m, 1H); 2.64-2.41 (m, 2H); 2.22 (m, 1H); 2.15-1.92 (m, 4H);
1.84-1.22 (m, 14H); 1.18 (t, J=7.2 Hz, 3H); 1.14-0.98 (m, 2H); 1.08
(s, 9H); 0.60-0.48 (m, 2H).
Example 60
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-(difluoromethyl)cyclopropyl]-18,18-difluoro-14-methoxy-3,6-dio-
xo-9-propyl-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-me-
thanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxal-
ine-8-carboxamide
##STR00308## ##STR00309## ##STR00310##
[0987] Step 1. Preparation of 60-1: To a solution of Intermediate
B5 (160 mg, 0.590 mmol) and Intermediate E3 (194 mg, 0.590 mmol) in
MeCN (2.95 mL) was added cesium carbonate (192 mg, 0.590 mmol) at
rt under an argon atmosphere. After 24 h, the reaction mixture was
then filtered through a pad of Celite and the filtrate concentrated
in vacuo. The crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes gradient) to afford
substituted quinoxaline 60-1. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.29H.sub.40F.sub.2N.sub.3O.sub.6: 564.28. found:
564.44.
[0988] Step 2. Preparation of 60-2: To a solution 60-1 (193 mg,
0.343 mmol) in tert-butyl acetate (1.36 mL) was added a solution of
methanesulfonic acid (111 .mu.L, 1.72 mmol) in dichloromethane
(0.34 mL) and the reaction was stirred at rt. After 2 h, the
reaction mixture was diluted with saturated sodium bicarbonate
solution (20 mL) and the resulting mixture was extracted with ethyl
acetate (2.times.20 mL). The combine organics were dried over
anhydrous sodium sulfate and were concentrated in vacuo to afford
amine hydrochloride 60-2, which was used directly in the next step
without further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.24H.sub.32F.sub.2N.sub.3O.sub.4: 464.23. found: 464.35.
[0989] Step 3. Preparation of 60-3: To a solution of 60-2 (133 mg,
0.289 mmol) and Intermediate D11 (133 mg, 0.412 mmol) in MeCN (1.7
mL) was added HATU (157 mg, 0.412 mmol) followed by DIPEA (298
.mu.L, 1.72 mmol) at rt under and argon atmosphere. After 1 h, the
reaction mixture was concentrated in vacuo, and the crude residue
was purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford amide 60-3. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.38H.sub.53F.sub.2N.sub.4O.sub.7:
715.38. found: 715.55.
[0990] Step 4. Preparation of 60-4: To a solution of 60-3 (188 mg,
264 .mu.mol) in DCE (52.8 mL) was added Zhan 1B catalyst (19.4 mg,
26.4 .mu.mol) and the reaction mixture was degassed for 10 minutes
with argon. The reaction mixture was then heated to 100.degree. C.
After 1 h, the reaction mixture was allowed to cool to rt and was
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (0-100% ethyl acetate/hexanes gradient) to afford
macrocycle 60-4. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49F.sub.2N.sub.4O.sub.7: 687.35. found: 687.54.
[0991] Step 5. Preparation of 60-5: To a solution of macrocycle
60-4 (119 mg, 173 .mu.mol) in ethanol (1.0 mL) was added Pd/C (10
wt %, 18.4 mg, 17.3 .mu.mol) at rt under an argon atmosphere. The
reaction vessel was evacuated and refilled with 1 atm hydrogen gas
(3.times.) and the reaction mixture was stirred vigorously at rt.
After 1 h, the reaction mixture was filtered through a pad of
Celite with ethyl acetate washings (3.times.2 mL). The filtrate was
concentrated in vacuo to afford macrocycle 60-5, which was used
directly in the next step without further purification. LCMS-ESI
(m/z): [M+H].sup.+ calcd for C.sub.36H.sub.51F.sub.2N.sub.4O.sub.7:
689.36. found: 689.56.
[0992] Step 6. Preparation of 60-6: To a solution of 60-5 (150 mg,
218 .mu.mol) in DCM (1.1 mL) was added TMSOTf (197 .mu.L, 1.09
mmol) at rt under an argon atmosphere. After 2 h, the reaction
mixture was transferred to a solution of 0.5N NaOH solution (5 mL)
precooled to 0.degree. C. The resulting mixture was acidified with
1N HCl solution to pH=2 and was extracted with dichloromethane
(3.times.5 mL). The combined organic extracts were dried over
anhydrous sodium sulfate and were concentrated in vacuo to afford
carboxylic acid 60-6, which was used directly in the next step
without further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.43F.sub.2N.sub.4O.sub.7: 633.30. found: 633.49.
[0993] Step 7. Preparation of Example 60: To a solution of 60-6
(100 mg, 158 .mu.mol) and Intermediate A9 (69.0 mg, 237 .mu.mol) in
MeCN (790 .mu.L) was added HATU (91.5 mg, 237 .mu.mol) followed by
DIPEA (137 .mu.L, 790 .mu.mol) at rt under an argon atmosphere.
After 3 h, the reaction mixture was concentrated in vacuo, was
purified by preparatory HPLC (Gemini 5u C18 110 .ANG. column,
5-100% MeCN/H.sub.2O, 0.1% trifluoroacetic acid modifier) and was
lyophilized to afford Example 60 as a TFA salt. Analytic HPLC
RetTime: 8.89 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.53F.sub.4N.sub.6O.sub.9S: 869.35. found: 859.66.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.29 (br s, 1H), 7.94 (d,
J=9.2 Hz, 1H), 7.31 (d, J=9.2 Hz, 1H), 7.19 (br s, 1H), 5.87 (br s,
1H), 5.84 (td, J.sub.H-F=55.8 Hz, J=5.4 Hz, 1H), 4.56 (d, J=6.9 Hz,
1H), 4.40 (d, J=12.6 Hz, 1H), 4.36 (s, 1H), 4.17 (dd, J=11.9, 3.4
Hz, 1H), 3.96 (br s, 4H), 3.68 (br s, 1H), 3.01-2.91 (m, 1H),
2.71-2.61 (m, 1H), 2.61-2.43 (m, 1H), 2.02 (br s, 4H), 1.88-1.59
(m, 4H), 1.59-1.35 (m, 4H), 1.33-1.20 (m, 3H), 1.09 (s, 9H),
1.04-0.95 (app t, J=7.0 Hz, 5H), 0.79-0.65 (m, 1H), 0.49 (d, J=6.5
Hz, 1H).
Example 61
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-2-(difluorometh-
yl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-10-ethyl-19,1-
9-difluoro-15-methoxy-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,2-
3b-hexadecahydro-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19-
][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00311##
[0995] Example 61 was prepared similarly to Example 59 substituting
Intermediate A10 for Intermediate A9 in Step 5. The TFA salt of
Example 61 was isolated. Analytic HPLC RetTime: 9.28 min. LCMS-ESI
(m/z): [M+H].sup.+ calcd for
C.sub.43H.sub.57F.sub.4N.sub.6O.sub.9S: 909.38. found: 909.59.
.sup.1H NMR (400 MHz, CD.sub.3OD) (9.28 (s, 1H); 7.95 (d, J=9.2 Hz,
1H); 7.33 (dd, J=9.2, 2.4 Hz, 1H); 7.23 (d, J=2.4 Hz, 1H); 6.0 (br
s, 1H); 5.83 (br s, 1H); 5.83 (td, J.sub.H-F=55 Hz, J=6 Hz, 1H);
4.94 (d, J=7.6 Hz, 1H); 4.61 (d, J=7.6 Hz, 1H); 4.34 (d, J=7.6 Hz,
1H); 4.32 (br s, 1H); 4.18 (m, 1H); 3.97 (s, 3H); 2.63-2.47 (m,
2H); 2.28-2.17 (m, 1H); 2.12-1.96 (m, 4H); 1.83-1.26 (m, 14H); 1.53
(s, 3H); 1.19 (t, J=7.2 Hz, 3H); 1.08 (s, 9H); 0.94-0.88 (m, 2H);
0.62-0.48 (m, 2H).
Example 62
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-1-[(cyclopropyl-
sulfonyl)carbamoyl]-2-(difluoromethyl)cyclopropyl]-19,19-difluoro-15-metho-
xy-10-methyl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexade-
cahydro-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,-
6]dioxadiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00312## ##STR00313##
[0997] Step 1. Preparation of Example 62-1: HATU (214 mg, 0.563
mmol, Oakwood) and DIPEA (0.30 mL, 1.72 mmol) were added to a
mixture of 46-2 (186 mg, 0.428 mmol) and Intermediate D16 (157 mg,
0.508 mmol) in 10 mL of acetonitrile under argon. After stirring
overnight, the reaction mixture was concentrated under reduced
pressure and the resulting residue was purified by silica gel
chromatography (0-30% ethyl acetate in hexanes) to yield
Intermediate 62-1. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.53F.sub.2N.sub.4O.sub.7: 727.38. found: 727.51.
[0998] Step 2. Preparation of 62-2: A mixture of 62-1 (275 mg,
0.378 mmol) and Zhan 1B catalyst (34 mg, 0.046 mmol, Strem) in 75
mL of DCE was deoxygenated with argon for 17 minutes. The mixture
was then heated at reflux for 80 minutes. An additional 8 mg of
Zhan 1B catalyst was added and mixture stirred at reflux for twenty
minutes. After cooling to room temperature, reaction mixture was
concentrated in vacuo. The resulting residue was purified by silica
gel chromatography (0-25% ethyl acetate in hexanes) to yield
intermediate 62-2. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.49F.sub.2N.sub.4O.sub.7: 699.35. found: 669.50.
[0999] Step 3. Preparation of mixture of 62-3: Palladium on carbon
(10 wt % Pd, 60 mg, 0.057 mmol) was added to a solution of 62-2
(207 mg, 0.297 mmol) in 7 mL of ethanol. The atmosphere was
replaced with hydrogen and mixture was stirred overnight. The
reaction was filtered over Celite, washing with ethanol. Filtrate
was concentrated in vacuo to yield intermediate 62-3, which was
used in the next step without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.37H.sub.51F.sub.2N.sub.4O.sub.7:
701.36. found: 701.65.
[1000] Step 4. Preparation of 62-4: TFA (1.6 mL, 20.9 mmol) was
added slowly to a solution of 62-3 (202 mg, 0.289 mmol) in 4.5 mL
of dichloromethane. After 3.5 hours, mixture was concentrated under
reduced pressure to near dryness. Resulting residue was taken up in
30 mL of ethyl acetate, washed with 20 mL of water, 20 mL of sat.
NaHCO.sub.3 (aq), and separated. Aqueous layers were extracted with
ethyl acetate (3.times.20 mL). Combined organics were washed with
30 mL of brine, dried over anhydrous MgSO.sub.4, filtered, and
concentrated in vacuo to yield intermediate 62-4, which was used in
the next step without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.33H.sub.43F.sub.2N.sub.4O.sub.7:
645.30. found: 645.53.
[1001] Step 5. Preparation of Example 62: HATU (113 mg, 0.297 mmol,
Oakwood) and DIPEA (0.17 mL, 0.978 mmol) were added to a mixture of
62-4 (120 mg, 0.186 mmol) and Intermediate A9 (110 mg, 0.379 mmol)
in 6 mL of acetonitrile under argon. After stirring for overnight,
reaction mixture was taken up in 30 mL of ethyl acetate and washed
with 20 mL of 1 N aqueous HCl. The aqueous layer was extracted
three times with ethyl acetate. Combined organics were washed with
50% brine, dried over anhydrous Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The resulting residue was purified by silica
gel chromatography (0-50% ethyl acetate in hexanes) and reverse
phase prep HPLC (50-100% acetonitrile in water, with 0.1%
trifluoroacetic acid buffer) to yield the trifluoroacetic acid salt
of Example 62. Analytic HPLC RetTime: 9.03 min. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.41H.sub.53F.sub.4N.sub.6O.sub.9S:
881.35. found: 881.57. .sup.1H NMR (400 MHz, CD.sub.3OD): .delta.
9.27 (s, 1H), 7.94 (d, J=8.8 Hz, 1H), 7.33 (dd, J=9.2, 2.8 Hz, 1H),
7.27 (d, J=2.8 Hz, 1H), 5.84 (td, J.sub.H-F=56 Hz, J=6.8 Hz, 1H),
5.75 (d, J=3.6 Hz, 1H), 4.94 (d, J=7.2 Hz, 1H), 4.55 (d, J=7.2 Hz,
1H), 4.35 (d, J=12 Hz, 1H), 4.32 (s, 1H), 4.22-4.16 (dd, J=12, 4
Hz, 1H), 3.97 (s, 3H), 3.01-2.94 (m, 1H), 2.81-2.72 (m, 1H),
2.66-2.40 (m, 1H), 2.36-2.28 (m, 1H), 2.10-1.94 (m, 4H), 1.82-1.72
(m, 2H), 1.70-1.22 (m, 10H), 1.14-1.02 (m, 7H), 1.10 (s, 9H),
0.61-0.49 (m, 2H).
Example 63
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18,18-difluoro--
14-methoxy-1a-methyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetrad-
ecahydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadeci-
no[11,12-b]quinoxaline-8-carboxamide
##STR00314## ##STR00315## ##STR00316##
[1003] Step 1. Preparation of 63-1: Amine hydrochloride 17-2 (500
mg, 1.03 mmol) was combined with intermediate mixture D17 (378.5
mg, 1.34 mmol), DIPEA (1.8 mL, 10.3 mmol) and DMF (3 mL). HATU
(587.1 mg, 1.55 mmol) was then added to the reaction mixture, which
was stirred at room temperature for 18 hrs. Reaction mixture was
then diluted with water (20 mL) and 1N HCl (10.5 mL) and taken up
into methylene chloride (20 mL). Organics were separated and
aqueous layer was extracted three times with methylene chloride (10
mL). Combined organics were then washed with brine, dried over
MgSO.sub.4, filtered, and concentrated in vacuo. Crude residue was
then purified via silica gel chromatography to give 63-1 as a 1:1
diastereomeric mixture. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.53F.sub.2N.sub.4O.sub.7: 715.4. found: 715.4.
[1004] Step 2. Preparation of 63-2 and 63-3: Diastereomeric mixture
63-1 (496 mg, 0.695 mmol) and Zhan 1B catalyst (53.8 mg, 0.0695
mmol, Strem) were dissolved in 140 mL of anhydrous DCE and sparged
with N.sub.2 for 30 minutes. The mixture was then heated to
100.degree. C. for 90 minutes, and an additional portion of Zhan 1B
catalyst was added (54 mg, 0.695 mmol, Strem). Reaction was then
cooled to room temperature and concentrated in vacuo. The resulting
residue was purified via silica gel chromatography (0% to 40% ethyl
acetate in hexanes) to yield single diastereomers 63-2 (early
eluting fraction) and 63-3 (late eluting fraction). Early eluting
fraction: LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49F.sub.2N.sub.4O.sub.7: 687.4. found: 687.2 Late
eluting fraction: LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49F.sub.2N.sub.4O.sub.7: 687.4. found: 687.3.
[1005] Step 3. Preparation of 63-4: Palladium on carbon (10% w/w,
155 mg) was added to a solution of 63-2 (155 mg, 0.226 mmol) in a
ethanol (3 mL). Mixture was stirred under an atmosphere of hydrogen
for 1 hr and was then filtered through a plug of Celite, and washed
with ethyl acetate. Filtrate was concentrated under reduced
pressure to yield 63-4, which was used in the next step without
further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.51F.sub.2N.sub.4O.sub.7: 689.4. found: 689.3.
[1006] Step 6. Preparation of 63-5: Intermediate 63-4 (153.5 mg,
0.222 mmol) was dissolved in a mixture of 1:1 TFA:DCM (6 mL) and
stirred at room temperature for 3 hrs. Reaction mixture was then
concentrated in vacuo to give 63-5, which was used in the
subsequent step without further purification. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.32H.sub.44N.sub.4O.sub.7: 633.3. found:
633.2.
[1007] Step 7. Preparation of Example 63: HATU (99.2 mg, 0.261
mmol) and DIPEA (271 .mu.L, 2.1 mmol) were added to a mixture of
63-5 (140.5 mg, 0.222 mmol) and A10 (100 mg, 0.316 mmol) in 1 mL of
DMF. After stirring overnight at room temperature, reaction mixture
was poured into water, acidified to pH 1 with 1 N aqueous HCl, and
extracted three times with methylene chloride (15 mL). Combined
organics were washed with water, brine, dried over MgSO.sub.4,
filtered, and concentrated under reduced pressure. The resulting
residue was purified by reverse phase prep HPLC (5-100%
acetonitrile in water, with 0.1% trifluoroacetic acid buffer) to
afford Example 63. Analytic HPLC RetTime: 8.951 min. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.55F.sub.4N.sub.6O.sub.9S: 883.4. found: 883.2.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.96 (d, J=9.2 Hz, 1H),
7.33 (dd, J=9.2, 2.8 Hz, 1H), 7.23 (d, J=2.7 Hz, 1H), 6.03 (d,
J=3.9 Hz, 1H), 5.80 (td, J=55.8, 6.7 Hz, 1H), 4.61 (d, J=6.9 Hz,
1H), 4.46 (d, J=12.2 Hz, 1H), 4.26-4.14 (m, 2H), 4.01-3.91 (m, 3H),
2.65-2.47 (m, 2H), 2.11-1.85 (m, 5H), 1.84-1.61 (m, 3H), 1.61-1.46
(m, 10H), 1.46-1.32 (m, 3H), 1.33-1.17 (m, 4H), 1.09 (d, J=15.9 Hz,
10H), 1.04-0.95 (m, 1H), 0.94-0.84 (m, 2H), 0.21-0.12 (m, 1H).
Example 64
Preparation of
(1aS,5S,8S,9S,10R,22aS)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18,18-difluoro--
14-methoxy-1a-methyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetrad-
ecahydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadeci-
no[11,12-b]quinoxaline-8-carboxamide
##STR00317##
[1009] Example 64 was prepared in a similar fashion to Example 63,
substituting late eluting 63-3 for early eluting 63-2 in Step 3.
Example 64 was then isolated. Analytic HPLC RetTime: 8.535 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S: 883.4. found: 883.3.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.97 (d, J=8.9 Hz, 1H),
7.45-7.16 (m, 2H), 5.97-5.52 (m, 2H), 4.74 (d, J=7.6 Hz, 1H),
4.50-4.16 (m, 1H), 4.06-3.86 (m, 5H), 2.77-2.57 (m, 1H), 2.51-2.18
(m, 2H), 2.16-1.86 (m, 5H), 1.75-1.32 (m, 16H), 1.33-1.03 (m, 14H),
1.02-0.76 (m, 2H), 0.42--0.09 (m, 1H).
Example 65
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-18,18-difluoro-14-metho-
xy-3,6-dioxo-9-propyl-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro--
8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12--
b]quinoxaline-8-carboxamide
##STR00318##
[1011] Step 1. Preparation of Example 65: To a solution of 60-6 (52
mg, 82 .mu.mol) and Intermediate A10 (37.5 mg, 123 .mu.mol) in MeCN
(411 .mu.L) was added HATU (47.5 mg, 123 .mu.mol) followed by DIPEA
(73 .mu.L, 411 .mu.mol) at rt under an argon atmosphere. After 20
h, the reaction mixture was concentrated in vacuo, was purified by
preparatory HPLC (Gemini 5u C18 110 .ANG. column, 5-100%
MeCN/H.sub.2O, 0.1% trifluoroacetic acid modifier) and was
lyophilized to afford Example 65 as a TFA salt. Analytic HPLC
RetTime: 8.99 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.55F.sub.4N.sub.6O.sub.9S: 883.36. found: 883.60.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.26 (s, 1H), 7.95 (d,
J=9.1 Hz, 1H), 7.33 (dd, J=9.2, 2.8 Hz, 1H), 7.22 (d, J=2.8 Hz,
1H), 5.89 (d, J=3.2 Hz, 1H), 5.81 (td, J.sub.H-F=55.5 Hz, J=6.5 Hz,
1H), 4.59 (d, J=7.0 Hz, 1H), 4.40 (d, J=12.5 Hz, 1H), 4.36 (s, 1H),
4.17 (dd, J=12.2, 3.8 Hz, 1H), 3.97 (s, 3H), 3.73-3.66 (m, 1H),
2.73-2.64 (m, 1H), 2.63-2.45 (m, 1H), 2.01 (br s, 3H), 1.85-1.62
(m, 4H), 1.62-1.53 (m, 3H), 1.51 (s, 3H), 1.48-1.22 (m, 5H), 1.08
(s, 9H), 1.01 (app t, J=7.3 Hz, 4H), 0.94-0.87 (m, 2H), 0.80-0.69
(m, 1H), 0.50 (d, J=7.1 Hz, 1H).
Example 66
Preparation of
(4aR,8S,11S,12S,13R,25aR)-8-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-(difluoromethyl)cyclopropyl]-12-ethyl-17-methoxy-6,9-dioxo-2-
,3,4,4a,6,7,8,9,12,13,21,22,23,24,25,25a-hexadecahydro-1 H,
11H-10,13-methanoquinoxalino[2,3-k][1,10,3,6]benzodioxadiazacyclononadeci-
ne-11-carboxamide
##STR00319## ##STR00320##
[1013] Step 1. Preparation of 66-1 and 66-2. To a solution of
Intermediate 70-3 (283 mg, 0.42 mmol) in CH.sub.2Cl.sub.2 (5 mL)
was added TMSOTf (380 .mu.L, 2.1 mmol). After stirring for 2 h, the
reaction mixture was poured into stirring 1 N NaOH (12 mL). The
mixture was transferred to a sept funnel, acidified to pH 3 with 1N
HCl, extracted with CH.sub.2Cl.sub.2, dried over magnesium sulfate,
and concentrated. The crude residue was purified by silica gel
chromatography (0-10% MeOH/EtOAc) to yield a mixture of 66-1 and
66-2. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.47N.sub.4O.sub.7: 623.34. found: 623.66.
[1014] Step 2. Preparation of 66-3 and 66-4. To a solution of 66-1
and 66-2 (58 mg, 0.09 mmol), intermediate A9 (32 mg, 0.11 mmol),
TBTU (42 mg, 0.13 mmol) and DMAP (16 mg, 0.14 mmol) in DMF (3 mL)
was added DIPEA (47 .mu.L, 0.27 mmol) and the reaction was stirred
at rt for 23 h. The reaction was quenched with water, diluted with
EtOAc, washed with sat. NaHCO.sub.3, brine, dried over magnesium
sulfate, and concentrated. The crude material was purified by
reverse phase HPLC (Gemini, 30-85% ACN/H.sub.2O+0.1% TFA) and
lyophilized to give the TFA salt of Intermediate 66-3 and 66-4
mixture. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.57F.sub.2N.sub.6O.sub.9S: 859.39. found: 859.65.
[1015] Step 3. Preparation of Example 66: To 66-3 and 66-4 (5 mg,
0.005 mmol) that was taken up in EtOH (2 mL) and treated with Pd/C
(10%, 5 mg). The atmosphere was replaced with hydrogen and stirred
at rt for 2.5 h. The reaction was filtered over Celite, washed with
EtOAc and concentrated. The residue was purified by silica gel
chromatography (0-10% MeOH/EtOAc) and lyophilized to give the
parent compound. Analytical HPLC RetTime: 9.15 min. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.59F.sub.2N.sub.6O.sub.9S: 862.01. found: 862.37.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.94-7.73 (m, 1H), 7.25
(m, 1H), 6.87 (d, J=9.8 Hz, 1H), 6.05 (m, 2H), 4.83-4.74 (m, 1H),
4.70 (d, J=7.6 Hz, 1H), 4.52-4.28 (m, 2H), 4.16 (m, 2H), 4.05-3.86
(m, 4H), 3.86-3.45 (m, 4H), 3.22-3.00 (m, 1H), 2.89 (s, 1H),
2.77-2.55 (m, 1H), 2.25 (t, J=7.3 Hz, 1H), 2.09-0.81 (m, 35H).
Example 67
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2S)-2-(2,2-difluoroethyl)-1-{-
[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-3-
,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-metha-
nocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-
-8-carboxamide
##STR00321##
[1017] Example 67 was prepared similarly to Example 1 substituting
Intermediate A8 for Intermediate A10 in Step 8. The TFA salt of
Example 67 was isolated. Analytic HPLC RetTime: 8.85 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S: 847.99. found: 847.64.
.sup.1H NMR (400 MHz, CD.sub.3OD) (9.00 (s, 1H); 7.79 (d, J=9.2 Hz,
1H); 7.23 (dd, J=9.2, 2.4 Hz, 1H); 7.15 (d, J=2.4 Hz, 1H); 5.89
(tt, J.sub.H-F=54 Hz, J=4.4 Hz, 1H); 5.89 (br s, 1H); 4.61 (d,
J=7.2 Hz, 1H); 4.39 (br s, 1H); 4.37 (d, J=9.2 Hz, 1H); 4.16 (dd,
J=9.2 Hz, 7.2 Hz, 1H); 3.92 (s, 3H); 3.78-3.72 (m, 1H); 3.10-2.88
(m, 1H); 2.86-2.74 (td, J=12, 4.4 Hz, 1H); 2.62-2.53 (m, 1H);
2.18-2.04 (m, 1H); 1.88-1.46 (m, 14H); 1.53 (s, 3H); 1.28-1.20 (m,
4H); 1.10 (s, 9H); 1.02-0.96 (m, 2H); 0.96-0.86 (m, 2H); 0.78-0.67
(m, 1H); 0.54-0.47 (m, 1H).
Example 68
Preparation of
(4aR,8S,11S,12S,13R,25aS)-8-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-(difluoromethyl)cyclopropyl]-21,21-difluoro-17-methoxy-12-me-
thyl-6,9-dioxo-2,3,4,4a,6,7,8,9,12,13,21,22,23,24,25,25a-hexadecahydro-1H,-
11H-10,13-methanoquinoxalino[2,3-k][1,10,3,6]benzodioxadiazacyclononadecin-
e-11-carboxamide
##STR00322## ##STR00323##
[1019] Step 1. Preparation of 68-1 and 68-2 (mixture): TMSOTf (0.6
mL, 3.3 mmol) was added to a solution of intermediate 62-3 (424 mg,
0.606 mmol) in 7 mL of dichloromethane at room temperature. After 1
hour, an additional 0.2 mL of TMSOTf was added. After a total of
three hours, reaction mixture was concentrated to yield a mixture
of 68-1 and 68-2 isomers, which was used in the next step without
further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.33H.sub.43F.sub.2N.sub.4O.sub.7: 645.30. found: 645.49.
[1020] Step 2. Preparation of 68-3 and 68-4 (mixture): HATU (209
mg, 0.550 mmol, Oakwood) and DIPEA (0.25 mL, 1.43 mmol) were added
to the mixture of 68-1 and 68-2 from the previous step (176 mg,
0.273 mmol) and Intermediate A9 (161 mg, 0.555 mmol) in 4 mL of
acetonitrile and 2 mL of DMF under argon. After one hour, an
additional 100 mg of Intermediate A9 was added. After two hours,
reaction mixture was taken up in 30 mL of ethyl acetate and washed
with 20 mL of 1 N aqueous HCl. The aqueous layer was extracted
three times with ethyl acetate. Combined organics were washed with
50% brine, dried over anhydrous Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The resulting residue was purified by silica
gel chromatography (0-50% ethyl acetate in hexanes) and reverse
phase prep HPLC (50-100% acetonitrile in water, with 0.1%
trifluoroacetic acid buffer) to yield the trifluoroacetic acid
salts of a mixture of 68-3 and 68-4. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.41H.sub.53F.sub.4N.sub.6O.sub.9S:
881.35. found: 881.50.
[1021] Step 3. Preparation of Example 68: Palladium on carbon (10
wt % Pd, 2 mg, 0.0019 mmol) was added to a solution of the mixture
of 68-3 and 68-4 from the previous step (4.5 mg, 0.0045 mmol) in 1
mL of ethanol. The atmosphere was replaced with hydrogen and
mixture was stirred for two hours. The reaction was filtered over
Celite, washing with ethanol. Filtrate was concentrated in vacuo to
yield Example 68. Analytic HPLC RetTime: 8.81 min. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.41H.sub.55F.sub.4N.sub.6O.sub.9S:
883.36. found: 883.64. .sup.1H NMR (400 MHz, CD.sub.3OD): .delta.
7.94 (d, J=10.4 Hz, 1H), 7.34-7.30 (m, 2H), 6.13 (td, J.sub.H-F=57
Hz, J=6.8 Hz, 1H), 5.88-5.84 (m, 1H), 4.62 (d, J=7.6 Hz, 1H),
4.38-4.30 (m, 2H), 4.20-4.05 (m, 2H), 3.98 (s, 3H), 2.87-2.76 (m,
2H), 2.34-2.16 (m, 2H), 1.92-1.54 (m, 6H), 1.46-1.36 (m, 3H),
1.34-1.12 (m, 8H), 1.20 (d, J=7.6 Hz, 3H), 1.08-0.96 (m, 4H), 1.04
(s, 9H), 0.93-0.78 (m, 4H).
Example 69
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-14-ethoxy-9-ethyl-3,6-d-
ioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocy-
clopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-c-
arboxamide
##STR00324## ##STR00325##
[1023] Step 1. Preparation of 69-2. Quinoxalinol 55-1 (54 mg, 0.086
mmol) was suspended in ACN (2 mL) and treated with Cs.sub.2CO.sub.3
(84 mg, 0.259 mmol) and bromoethane (0.032 mL, 0.432 mmol). The
reaction mixture was stirred at RT for 16 h. The reaction was
filtered and the crude material was purified by flash column
chromatography to afford 69-2. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.36H.sub.52N.sub.4O.sub.7: 652.38. found: 653.41.
[1024] Step 2. Preparation of 69-3. Intermediate 69-2 (0.086 mmol
theoretical) was treated with DCM (10 mL) and TMSOTf (1.0 mL) at
RT. After 1 h, the reaction was complete determined by LCMS. The
reaction was concentrated under reduced pressure to afford 69-3,
which was carried on without further purification. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.32H.sub.44N.sub.4O.sub.7: 596.32.
found: 597.38.
[1025] Step 3. Preparation of Example 69. Carboxcylic acid 69-3
(0.0.086 mmol theoretical) was treated with intermediate A10 (40
mg, 0.130 mmol), TBTU (47 mg, 0.147 mmol), DMAP (18 mg, 0.147
mmol), DCM (3 mL) and DIPEA (0.075 mL, 0.432 mmol). The reaction
mixture was stirred at RT for 20 h, then concentrated under reduced
pressure and purified by reverse phase HPLC to afford Example 69 as
a TFA salt. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.56F.sub.2N.sub.6O.sub.9S: 846.38. found: 847.75.
Example 70
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-1-[(cyclopropyl-
sulfonyl)carbamoyl]-2-(difluoromethyl)cyclopropyl]-10-ethyl-15-methoxy-4,7-
-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadecahydro-1H,9H-8,-
11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6]dioxadiazacycl-
ononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00326## ##STR00327##
[1027] Step 1. Preparation of 70-1: To a solution of 1-2 (575 mg,
1.41 mmol), D12 (410 mg, 1.26 mmol) and HATU (696 mg, 1.80 mmol) in
DMF (12 mL) was added DIPEA (1.0 mL, 5.64 mmol) and the reaction
was stirred at rt. After stirring for 2 h, additional HATU (350 mg,
0.92 mmol) and DIPEA (0.5 mL, 2.8 mmol) was added to the reaction,
and the mixture was stirred for 14 h. The reaction was quenched
with sat. NaHCO.sub.3 solution and extracted with EtOAc, washed
subsequently with brine, dried over magnesium sulfate and
concentrated. The crude product was purified by silica gel
chromatography (10-30% EtOAc/hexanes) to yield intermediate 70-1.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.54ClN.sub.4O.sub.7: 713.37. found: 713.95.
[1028] Step 2. Preparation of 70-2: To a solution of 70-1 (542 mg,
0.76 mmol), TEA (0.16 mL, 1.14 mmol) and potassium
vinyltrifluoroborate (153 mg, 1.14 mmol) in EtOH (10 mL) was added
PdCl.sub.2(dppf) (62 mg, 0.08 mmol). The reaction was degassed with
N.sub.2 for 10 min and heated to 80.degree. C. for 1 h. The
reaction was quenched with sat. NaHCO.sub.3 solution and extracted
with EtOAc, washed subsequently with brine, dried over magnesium
sulfate and concentrated. The residue was purified using silica gel
chromatography (0-20% EtOAc/hexanes) to give intermediate 70-2.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.57N.sub.4O.sub.7: 705.42. found: 705.05.
[1029] Step 3 and 4. Preparation of 70-3: To a solution of 70-2
(470 mg, 0.66 mmol) in DCE (100 mL) was added Zhan 1B catalyst (49
mg, 0.07 mmol) and the reaction was degassed for 30 minutes with
N.sub.2. The reaction was heated to 100.degree. C. for 1 h, allowed
to cool to rt and concentrated. The crude product was purified by
silica gel chromatography to give product (358 mg; LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for C.sub.38H.sub.53N.sub.4O.sub.7:
677.39. found: 677.52) that was taken up in EtOH (6 mL) and EtOAc
(2 mL) and treated with Pd/C (10%, 350 mg). The atmosphere was
replaced with hydrogen and stirred at rt for 1.5 h. The reaction
was filtered over Celite, washed with EtOAc and concentrated (358
mg intermediate 70-3) that was used subsequently without further
purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.55N.sub.4O.sub.7: 679.41. found: 679.44.
[1030] Step 5. Preparation of 70-4: To a solution of 70-3 (100 mg,
0.15 mmol) in DCM (1 mL) was added TFA (1 mL) and stirred at rt for
2 h. The reaction was diluted with EtOAc, washed with H.sub.2O,
basicified to pH 7 with sat. NaHCO.sub.3 solution, dried over
magnesium sulfate, and concentrated to give a residue of
intermediate 70-4 that was used subsequently without further
purification LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.47N.sub.4O.sub.7: 623.34. found: 623.44.
[1031] Step 6. Preparation of Example 70: To a solution of 70-4 (94
mg, 0.15 mmol), intermediate A9 (65 mg, 0.22 mmol), TBTU (87 mg,
0.27 mmol) and DMAP (27 mg, 0.22 mmol) in DCM (3 mL) was added
DIPEA (0.13 mL, 0.75 mmol) and the reaction was stirred at rt for 2
h. The reaction was quenched with water, diluted with EtOAc, washed
with sat. NaHCO.sub.3, brine, dried over magnesium sulfate, and
concentrated. The crude material was purified by reverse phase HPLC
(Gemini, 30-85% ACN/H.sub.2O+0.1% TFA) and lyophilized to give
Example 70 (23 mg) as a TFA salt.
[1032] Analytical HPLC RetTime: 9.32 min. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.42H.sub.57F.sub.2N.sub.6O.sub.9S:
859.39. found: 859.54. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
9.31 (s, 1H), 7.83 (d, J=9.1 Hz, 1H), 7.26 (dd, J=9.1, 2.8 Hz, 1H),
7.20 (d, J=2.7 Hz, 1H), 6.09-5.68 (m, 2H), 5.51 (s, 1H), 5.07-4.97
(m, 1H), 4.70-4.55 (m, 1H), 4.42-4.29 (m, 2H), 4.22 (dd, J=12.0,
4.1 Hz, 1H), 3.96 (s, 2H), 3.75 (t, J=6.7 Hz, 2H), 3.02 (m, 2H),
2.93-2.67 (m, 1H), 2.56 (m, 1H), 2.13-1.04 (m, 30H), 1.00 (d, J=6.6
Hz, 1H), 0.90 (m, 3H), 0.65-0.46 (m, 2H).
Example 71
Preparation of
(4aR,8S,11S,12S,13R,25aR)-8-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-(difluoromethyl)cyclopropyl]-17-methoxy-12-methyl-6,9-dioxo--
2,3,4,4a,6,7,8,9,12,13,21,22,23,24,25,25a-hexadecahydro-1H,11H-1,3:10,13-d-
imethanoquinoxalino[2,3-k][1,10,3,6]benzodioxadiazacyclononadecine-11-carb-
oxamide and
##STR00328## ##STR00329##
[1034] Step 1: To a solution of amine 18-2 (315 mg, 0.80 mmol),
DIPEA (350 .mu.L, 2.0 mmol) and a 1:1 mixture of acids D19 (270 mg,
0.80 mmol) in MeCN (8 mL) was added HATU (400 mg, 1.05 mmol). The
resulting solution was stirred for 2.5 h at r.t., at which time it
was diluted with EtOAc (50 mL) and 0.2 N aqueous HCl (30 mL). The
phases were separated, and the organic phase was dried over
MgSO.sub.4, filtered, and concentrated to afford a crude residue.
Purification by silica gel chromatography (10% to 30% EtOAc in
hexanes) provided 474 mg of a colorless oil that was used directly
in the next step.
[1035] Step 2: A suspension of the product from step 1 (474 mg, ca.
0.65 mmol), PdCl.sub.2(dppf)*CH.sub.2Cl.sub.2 (40 mg, 0.049 mmol)
and potassium vinyltrifluoroborate (189 mg, 1.41 mmol) in EtOH (8
mL) was sparged with Ar for several minutes and Et.sub.3N (200
.mu.L, 1.4 mmol) was added. The resulting mixture was heated under
Ar to 75.degree. C. via oil bath. After stirring 2.25 h, the
reaction mixture was cooled to r.t. and was diluted with EtOAc (35
mL) and half-saturated brine (20 mL). The phases were separated,
and the organic phase was dried over Na.sub.2SO.sub.4, filtered,
and concentrated to afford a crude residue. Purification by silica
gel chromatography provided a yellow oil that was used directly in
the next step.
[1036] Step 3: A solution of the product from Step 3 (395 mg, 0.56
mmol) in 1,2-DCE (180 mL) was sparged with Ar for 10 min. Zhan 1B
metathesis catalyst (61 mg, 0.083 mmol) was then added as a
solution in DCE (4 mL), and the resulting solution was heated to
85.degree. C. After stirring 1.75 h, the reaction mixture was
cooled to ambient temperature, concentrated onto silica gel (5 g),
and purified by silica gel chromatography (10 to 15 to 25% EtOAc in
hexanes) to afford 116 mg of a fast-eluting product and 84 mg of a
slow-eluting product.
[1037] Step 4-5 (fast-eluting diastereomer): The fast-eluting
product from Step 3 was dissolved in 1:1 EtOAc:EtOH (4 mL). Pd/C
(10 wt. % Pd, 45 mg) was added, and the reaction vessel was purged
twice with 1 atm H.sub.2. The reaction mixture was stirred for 2.5
h under 1 atm H.sub.2 and was then filtered through celite with
EtOAc to afford a crude residue. This residue was dissolved in
CH.sub.2Cl.sub.2 (1 mL) and was treated with TFA (2 mL). After
stirring 2 h, the reaction mixture was concentrated in vacuo and
was partitioned between EtOAc (15 mL) and 15% saturated aqueous
NaHCO.sub.3 (10 mL). The phases were separated, and the organic
phase was washed with brine (10 mL), dried over Na.sub.2SO.sub.4,
and filtered to provide 71-1. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.47N.sub.4O.sub.7: 623.3. found: 623.2.
[1038] Step 4-5 (slow-eluting diastereomer): The slow-eluting
product from Step 3 was dissolved in EtOAc (1 mL) and EtOH (7 mL).
Pd/C (10 wt. % Pd, 85 mg) was added, and the reaction vessel was
purged twice with 1 atm H.sub.2. The reaction mixture was stirred
for 3 h under 1 atm H.sub.2 and was then filtered through celite
with EtOAc to afford a crude residue. This residue was dissolved in
CH.sub.2Cl.sub.2 (1 mL) and was treated with TFA (2 mL). After
stirring 2 h, the reaction mixture was concentrated in vacuo and
was partitioned between EtOAc (15 mL) and 15% saturated aqueous
NaHCO.sub.3 (10 mL). The phases were separated, and the organic
phase was washed with brine (10 mL), dried over Na.sub.2SO.sub.4,
and filtered to provide 71-2. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.34H.sub.47N.sub.4O.sub.7: 623.3. found: 623.2.
[1039] Step 6: Preparation of Example 71: To a suspension of acid
71-1 (49 mg, 0.079 mmol) and amine hydrochloride A9 (41 mg, 0.14
mmol) in MeCN (1 mL) was added DIPEA (100 .mu.L, 0.57 mmol). HATU
(45 mg, 0.12 mmol) was added to the resulting solution, and the
reaction was stirred at rt for 14.5 h. The reaction was then
diluted with EtOAc (20 mL), 0.2 M aqueous HCl (10 mL) and brine (10
mL). The phases were separated and the aqueous phase was extracted
with EtOAc (20 mL). The combined organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue. This residue was dissolved in CH.sub.2Cl.sub.2 and was
concentrated onto 2 g silica gel. Purification by silica gel
chromatography (4% to 45% acetone in hexanes) provided an amorphous
residue that was lyophilized from water and MeCN to provide Example
71. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.57F.sub.2N.sub.6O.sub.9S: 859.4. found: 859.1.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.13 (s, 1H), 7.81 (d,
J=9.1 Hz, 1H), 7.63 (s, 1H), 7.19 (dd, J=9.1, 2.8 Hz, 1H), 7.09 (d,
J=2.7 Hz, 1H), 5.97 (td, J=55.5, 6.9 Hz, 1H), 5.59-5.45 (m, 2H),
4.96 (dd, J=14.4, 6.2 Hz, 1H), 4.51 (d, J=7.2 Hz, 1H), 4.42 (d,
J=9.8 Hz, 1H), 4.13 (dt, J=12.0, 7.7 Hz, 2H), 3.93 (s, 3H),
2.99-2.63 (m, 4H), 2.40-2.23 (m, 2H), 2.15-0.83 (m, 34H).
Example 72
Preparation of
(4aS,8S,11S,12S,13R,25aS)-8-tert-butyl-N-[(1R,2R)-1-[(cyclopropylsulfonyl-
)carbamoyl]-2-(difluoromethyl)cyclopropyl]-17-methoxy-12-methyl-6,9-dioxo--
2,3,4,4a,6,7,8,9,12,13,21,22,23,24,25,25a-hexadecahydro-1H,11H-1,3:10,13-d-
imethanoquinoxalino[2,3-k][1,10,3,6]benzodioxadiazacyclononadecine-11-carb-
oxamide
##STR00330##
[1041] Step 1: Preparation of Example 72: To a suspension of acid
71-2 (49 mg, 0.079 mmol) and amine hydrochloride A9 (38 mg, 0.13
mmol) in MeCN (1 mL) was added DIPEA (100 .mu.L, 0.57 mmol). HATU
(41 mg, 0.11 mmol) was added to the resulting solution, and the
reaction was stirred at rt for 14.5 h. The reaction was then
diluted with EtOAc (20 mL), 0.2 M aqueous HCl (10 mL) and brine (10
mL). The phases were separated and the aqueous phase was extracted
with EtOAc (20 mL). The combined organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated to afford a crude
residue. This residue was dissolved in CH.sub.2Cl.sub.2 and was
concentrated onto 2 g silica gel. Purification by silica gel
chromatography (4% to 45% acetone in hexanes) provided an amorphous
residue that was lyophilized from water and MeCN to provide Example
72. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.57F.sub.2N.sub.6O.sub.9S: 859.4. found: 859.0.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.72 (s, 1H), 9.36 (s,
1H), 7.86 (d, J=9.1 Hz, 1H), 7.28 (d, J=2.7 Hz, 1H), 7.25-7.17 (m,
2H), 5.98-5.88 (m, 1H), 5.69 (td, J=55.4, 6.9 Hz, 1H), 4.81-4.69
(m, 1H), 4.68-4.56 (m, 2H), 4.33 (d, J=10.1 Hz, 1H), 3.99 (s, 3H),
3.35 (dd, J=9.7, 7.0 Hz, 1H), 3.24-3.13 (m, 1H), 2.97-2.87 (m, 1H),
2.87-2.72 (m, 2H), 2.57-2.45 (m, 1H), 2.38-2.28 (m, 1H), 2.17-0.71
(m, 34H).
Example 73
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-9-ethyl-14-methoxy-N-[(1R,2R)-2-meth-
yl-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-3,6-dioxo-1,1a-
,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[-
18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamid-
e
##STR00331##
[1043] Example 73 was prepared similarly to Example 1 substituting
Intermediate A11 for Intermediate A10 in Step 8. The TFA salt of
Example 73 was isolated. Analytic HPLC RetTime: 8.72 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.57N.sub.6O.sub.9S: 797.98. found: 797.54. .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.84 (s, 1H); 7.79 (d, J=9.2 Hz, 1H);
7.22 (dd, J=9.2, 2.4 Hz, 1H); 7.13 (d, J=2.4 Hz, 1H); 5.87 (d,
J=3.2 Hz, 1H); 4.57 (d, J=7.2 Hz, 1H); 4.39 (br s, 1H); 4.37 (br d,
J=10 Hz, 1H); 4.15 (dd, J=12, 4 Hz, 1H); 3.92 (s, 3H); 3.74 (m,
1H); 3.10-2.88 (m, 1H); 2.80 (td, J=12.4, 4 Hz, 1H); 2.58 (m, 1H);
1.89-1.66 (m, 3H); 1.66-1.38 (m, 11H); 1.52 (s, 3H); 1.23 (t, J=7.2
Hz, 3H); 1.16 (d, J=6 Hz, 3H); 1.10 (s, 9H); 1.02-0.84 (m, 4H);
0.78-0.66 (m, 1H); 0.55-0.20 (m, 1H).
Example 74
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-fluorocyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-14-methoxy-3,6--
dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanoc-
yclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8--
carboxamide
##STR00332##
[1045] Example 74 was prepared similarly to Example 1 substituting
Intermediate A12 for Intermediate A10 in Step 8. The TFA salt of
Example 74 was isolated. Analytic HPLC RetTime: 8.81 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.52F.sub.3N.sub.6O.sub.9S: 837.35. found: 837.54.
.sup.1H NMR (400 MHz, CD.sub.3OD) (9.26 (s, 1H); 7.79 (d, J=9.2 Hz,
1H); 7.22 (dd, J=9.2, 2.4 Hz, 1H); 7.14 (d, J=2.4 Hz, 1H); 5.89 (d,
J=3.6 Hz, 1H); 5.82 (td, J.sub.H-F=56 Hz, J=6.4 Hz, 1H); 4.56, (d,
J=7.2 Hz, 1H); 4.39 (s, 1H); 4.38 (d, J=12 Hz, 1H); 4.16 (dd, J=12,
7.2 Hz, 1H); 3.92 (s, 3H); 3.78-3.72 (m, 1H); 3.10-2.89 (m, 1H);
2.80 (td, J=12, 4 Hz, 1H); 2.63-2.54 (m, 1H); 2.02 (m, 2H);
1.95-1.66 (m, 3H); 1.66-1.36 (m, 9H); 1.22 (t, J=7.2 Hz, 3H);
1.14-1.04 (m, 2H); 1.09 (s, 9H); 1.04-0.92 (m, 2H); 0.78-0.68 (m,
1H); 0.57-0.46 (m, 1H).
Example 75
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-1-{[(1-chlorocyclopropyl)-
sulfonyl]carbamoyl}-2-(difluoromethyl)cyclopropyl]-9-ethyl-14-methoxy-3,6--
dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanoc-
yclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8--
carboxamide
##STR00333##
[1047] Example 75 was prepared similarly to Example 1 substituting
Intermediate A13 for Intermediate A10 in Step 8. The TFA salt of
Example 75 was isolated. Analytic HPLC RetTime: 8.89 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.52ClF.sub.2N.sub.6O.sub.9S: 853.32. found: 853.94.
.sup.1H NMR (400 MHz, CD.sub.3OD) (9.24 (s, 1H); 7.79 (d, J=9.2 Hz,
1H); 7.22 (dd, J=9.2, 2.4 Hz, 1H); 7.13 (d, J=2.4 Hz, 1H); 5.88 (d,
J=3.2 Hz, 1H); 5.84 (td, J.sub.H-F=55.6 Hz, J=6.8 Hz, 1H); 4.57 (d,
J=7.2 Hz, 1H); 4.39 (br s, 1H); 4.38 (d, J=12 Hz, 1H); 4.16 (dd,
J=12, 7.2 Hz, 1H); 3.92 (s, 3H); 3.77-3.73 (m, 1H); 3.00-2.88 (m,
1H); 2.86-2.75 (m, 1H); 2.64-2.54 (m, 1H); 2.10-1.90 (m, 4H);
1.90-1.37 (m, 12H); 1.23 (t, J=7.2 Hz, 3H); 1.10 (s, 9H); 1.02-0.96
(m, 2H); 0.78-0.64 (m, 1H); 0.56-0.45 (m, 1H).
Example 76
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-18,18-difluoro-14-metho-
xy-1a,9-dimethyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecah-
ydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[1-
1,12-b]quinoxaline-8-carboxamide
##STR00334## ##STR00335## ##STR00336##
[1049] Step 1. Preparation of 76-1: HATU (502 mg, 1.32 mmol,
Oakwood) and DIPEA (0.70 mL, 4.02 mmol) were added to a mixture of
46-2 (434 mg, 0.998 mmol) and Intermediate D17 (350 mg, 1.24 mmol)
in 16 mL of acetonitrile under argon. After stirring overnight, the
reaction mixture was concentrated under reduced pressure and the
resulting residue was purified by silica gel chromatography (0-25%
ethyl acetate in hexanes) to yield 76-1. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.37H.sub.51F.sub.2N.sub.4O.sub.7:
701.36. found: 701.57.
[1050] Step 2. Preparation of 76-2 and 76-3: A diastereomeric
mixture 76-1 (550 mg, 0.786 mmol) and Zhan 1B catalyst (69 mg,
0.094 mmol, Strem) in 157 mL of DCE was deoxygenated under argon
for 25 minutes. The mixture was then heated at reflux for 90
minutes. An additional 35 mg of Zhan 1B catalyst was added and
reaction mixture was heated at reflux for 45 minutes. After cooling
to room temperature, reaction mixture was concentrated in vacuo.
The resulting residue was purified by silica gel chromatography
(0-35% ethyl acetate in hexanes) to yield single diastereomers 76-2
(early eluting component) as a white solid film and 76-3 (late
eluting component) as a brown solid film. Early eluting 76-2:
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.47F.sub.2N.sub.4O.sub.7: 673.33. found: 673.45. Late
eluting 76-3: LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.47F.sub.2N.sub.4O.sub.7: 673.33. found: 673.47.
[1051] Step 3. Preparation of 76-4: Palladium on carbon (10 wt %
Pd, 51 mg, 0.048 mmol) was added to a solution of 76-2 (175 mg,
0.260 mmol) in 9 mL of ethanol. The atmosphere was replaced with
hydrogen and the reaction stirred overnight. The reaction mixture
was filtered over Celite and washed with ethanol. Filtrate was
concentrated in vacuo to yield 76-4, which was used in the next
step without further purification. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.35H.sub.49F.sub.2N.sub.4O.sub.7:
675.35. found: 675.53.
[1052] Step 4. Preparation of 76-5: TFA (1.2 mL, 15.6 mmol) was
added slowly to a solution of 76-4 (155 mg, 0.230 mmol) in 3.4 mL
of dichloromethane. After 4 hours, mixture was concentrated under
reduced pressure to near dryness. Resulting residue was taken up in
25 mL of ethyl acetate, washed with 15 mL of water, 15 mL of sat.
NaHCO.sub.3 (aq), and separated. Aqueous layers were extracted with
ethyl acetate (3.times.20 mL). Combined organics were washed with
30 mL of brine, dried over anhydrous MgSO.sub.4, filtered, and
concentrated in vacuo to yield 76-5, which was used in the next
step without further purification. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.31H.sub.41F.sub.2N.sub.4O.sub.7:
619.29. found: 619.44.
[1053] Step 5. Preparation of Example 76: HATU (160 mg, 0.421 mmol)
and DIPEA (0.20 mL, 1.15 mmol) were added to a mixture of 76-5 (140
mg, 0.226 mmol) and Intermediate A10 (139 mg, 0.457 mmol) in 7.5 mL
of MeCN under argon. After stirring for overnight, reaction mixture
was taken up in 30 mL of ethyl acetate and washed with 20 mL of 1 N
aqueous HCl. Layers were separated and aqueous was extracted three
times with ethyl acetate. Combined organics were washed with brine,
dried over anhydrous MgSO.sub.4, filtered, and concentrated in
vacuo. The resulting residue was purified by silica gel
chromatography (0-45% ethyl acetate in hexanes) and reverse phase
prep HPLC (50-100% acetonitrile in water, with 0.1% trifluoroacetic
acid buffer) to yield the trifluoroacetic acid salt of Example 76
(. Analytic HPLC RetTime: 8.80 min. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.40H.sub.53F.sub.4N.sub.6O.sub.9S: 869.35. found:
869.59. .sup.1H NMR (400 MHz, CD.sub.3OD): 9.19 (s, 1H), 7.94 (d,
J=9.2 Hz, 1H), 7.32 (dd, J=9.2, 2.8 Hz, 1H), 7.27 (d, J=2.8 Hz,
1H), 5.78 (td, J.sub.H-F=56 Hz, J=7.2 Hz, 1H), 5.76-5.74 (m, 1H),
4.56 (d, J=6.4 Hz, 1H), 4.48 (d, J=12 Hz, 1H), 4.27-4.19 (m, 1H),
4.22 (s, 1H), 3.97 (s, 3H), 2.76-2.70 (m, 1H), 2.62-2.43 (m, 1H),
2.14-1.94 (m, 3H), 1.90-1.80 (m, 1H), 1.80-1.62 (m, 3H), 1.56-1.52
(m, 2H), 1.51 (s, 3H), 1.49 (s, 3H), 1.41-1.36 (m, 1H), 1.27-1.18
(m, 1H), 1.11 (s, 9H), 1.09-1.04 (m, 5H), 1.03-0.94 (m, 2H),
0.87-0.81 (m, 3H), 0.17-0.12 (m, 1H).
Example 77
Preparation of
(1aS,5S,8S,9S,10R,22aS)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-18,18-difluoro-14-metho-
xy-1a,9-dimethyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecah-
ydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[1-
1,12-b]quinoxaline-8-carboxamide
##STR00337##
[1055] Example 77 was prepared in a similar fashion to Example 76,
substituting late eluting 76-3 for early eluting 76-2 in step 3.
Example 76 was then isolated. Analytic HPLC RetTime: 8.46 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.53F.sub.4N.sub.6O.sub.9S: 869.35. found: 869.53.
.sup.1H NMR (400 MHz, CD.sub.3OD): 7.95 (d, J=8.8 Hz, 1H), 7.32 (d,
J=8.8 Hz, 1H), 7.28 (s, 1H), 6.58-6.54 (m, 1H), 5.75 (td,
J.sub.H-F=55 Hz, J=6.8 Hz, 1H), 5.54-5.50 (m, 1H), 4.65 (d, J=6.8
Hz, 1H), 4.46 (d, J=12.8 Hz, 1H), 4.26-4.18 (m, 1H), 3.97 (s, 3H),
2.92-2.71 (m, 1H), 2.50-1.94 (m, 6H), 1.68-1.57 (m, 2H), 1.56-1.52
(m, 2H), 1.51 (s, 3H), 1.50-1.47 (m, 1H), 1.46-1.38 (m, 3H), 1.44
(s, 3H), 1.27-1.18 (m, 2H), 1.17-1.01 (m, 3H), 1.09 (s, 9H),
0.94-0.82 (m, 4H), 0.17-0.12 (m, 1H).
Example 78
Preparation of
(1aR,5S,8S,9S,10R,19E,22aR)-5-tert-butyl-14-cyano-N-[(1R,2R)-2-(difluorom-
ethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18-
,18-difluoro-3,6-dioxo-1,1a,3,4,5,6,9,10,17,17a,18,21,22,22a-tetradecahydr-
o-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,1-
2-b]quinoxaline-8-carboxamide
##STR00338## ##STR00339## ##STR00340##
[1057] Steps 1-4. Intermediate 78-4 was prepared similarly to
Intermediate 17-4, using E6 in place of E3.
[1058] Step 5: To a solution of 78-4 (90 mg, 0.135 mmol) in EtOH
(0.7 mL) was added NaBH.sub.4 (21 mg, 0.54 mmol). The reaction
mixture was stirred at rt for 1 h. After which time the reaction
mixture was filtered through a pad of celite and concentrated to
give intermediate 78-5, which was used subsequently without further
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.45F.sub.2N.sub.5O.sub.6: 669.76. found: 669.73.
[1059] Steps 6 and 7: Preparation of Example 78: To a solution of
78-5 (35 mg, 0.31 mmol) in DCM (0.4 mL), TFA (0.2 mL) was added and
the mixture was stirred at 20.degree. C. for 3 h. Solvents were
removed in vacuo to afford a residue was used subsequently without
further purification. To a suspension of this residue (33 mg, 0.05
mmol) and Intermediate A10 (27 mg, 0.1 mmol) in DCM (0.3 mL) was
added TBTU (26 mg, 0.08 mmol) and DIPEA (35 .mu.L, 0.2 mmol) at rt.
After 1 h, the solution was directly purified by reverse phase HPLC
(Gemini 5u C.sub.18 110 .ANG. column, 50-100% ACN/H.sub.2O+0.1%
TFA) and lyophilized to afford the TFA salt of Example 78.
Analytical HPLC RetTime: 7.994 min. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.40H.sub.49F.sub.4N.sub.7O.sub.8S: 863.92. found:
864.20. .sup.1H NMR (400 MHz, CD3OD) .delta. 9.35 (s, 1H), 7.29 (d,
1H), 7.18 (dd, 1H), 6.64 (d, 1H), 6.01-5.82 (m, 2H), 5.41 (m, 2H),
4.57-4.07 (m, 5H), 3.52 (m, 1H), 2.55-2.28 (m, 2H), 2.06-1.98 (m,
2H), 1.85 (m, 1H), 1.69-1.37 (m, 9H), 1.33 (m, 2H), 1.06-0.87 (m,
16H), 0.70 (m, 2H)., 0.49 (m, 1H).
Example 79
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-15-chloro-N-[(1R,2R)-1-[(c-
yclopropylsulfonyl)carbamoyl]-2-(difluoromethyl)cyclopropyl]-10-methyl-4,7-
-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadecahydro-1H,9H-8,-
11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6]dioxadiazacycl-
ononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00341## ##STR00342## ##STR00343##
[1061] Step 1. Preparation of 79-1. Sulfonyl quinoxaline E5 (920
mg, 3.32 mmol) was suspended in MeCN (17 mL), then treated with
intermediate B1 (1.00 g, 3.32 mmol) and Cs.sub.2CO.sub.3. After 17
h, the reaction mixture was filtered over celite and concentrated
under reduced pressure. The crude residue was purified by silica
column chromatography (10% to 30% EtOAc/Hex) to afford ether 79-1.
LCMS-ESI.sup.+ (m/z): [M-Boc+2H].sup.+ calcd for
O.sub.18H.sub.22Cl.sub.2N.sub.3O.sub.3: 398.10. found: 398.12.
[1062] Step 2. Preparation of 79-2. tert-Butyl carbamate 79-1 (513
mg, 1.03 mmol) was dissolved in DCM (10 mL) and treated with HCl
(4.0 mL in dioxane, 5 mL, 20 mmol). The reaction mixture was
stirred at RT for 1.5 h, then concentrated under reduced pressure
to afford amine hydrochloride 79-2, which was carried on without
purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.18H.sub.22Cl.sub.2N.sub.3O.sub.3: 398.10. found: 398.16.
[1063] Step 3. Preparation of 79-3. Amine hydrochloride 79-2 (1.03
mmol theoretical) and intermediate D12 (336 mg, 1.04 mmol) were
combined and treated with BEP (285 mg, 1.04 mmol), EtOAc (9 mL),
NMP (1 mL) and DIPEA (0.90 mL, 5.2 mmol). The reaction mixture was
stirred at 50.degree. C. for 3 h, then cooled to RT. After an
additional 15 h, the reaction mixture was diluted with EtOAc. The
organic solution was washed with saturated aqueous NaHCO.sub.3 and
brine, then dried over MgSO.sub.4, filtered and concentrated under
reduced pressure. The crude residue was purified by silica column
chromatography (10% to 25% EtOAc/Hex) to afford amide 79-3.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49Cl.sub.2N.sub.4O.sub.6: 703.30. found: 703.91.
[1064] Step 4. Preparation of 79-4. Chloro quinoxaline 79-3 (541
mg, 0.769 mmol) was treated with potassium vinyltrifluoroborate
(154 mg, 1.15 mmol), Pd(dppf)Cl.sub.2 dichloromethane adduct (63
mg, 0.077 mmol), EtOH (8 mL) and triethylamine (0.16 mL, 1.15
mmol). The stirred mixture was heated to reflux for 1 h, then
cooled to RT and diluted with EtOAc. The organic solution was
washed with saturated aqueous NaHCO.sub.3 and brine, then dried
over MgSO.sub.4, filtered and concentrated under reduced pressure.
The crude residue was purified by silica column chromatography (10%
to 30% EtOAc/Hex) to afford vinyl quinoxaline 79-4. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.38H.sub.52ClN.sub.4O.sub.6: 695.36.
found: 695.10.
[1065] Step 5. Preparation of 79-5. Vinyl quinoxaline 79-4 (390 mg,
0.561 mmol) was treated with DCE (112 mL) and Zhan-B catalyst (38
mg, 0.0561 mmol). The stirred mixture was degassed with bubbling
N.sub.2 for 25 min, then heated to reflux under an Ar atmosphere.
After 1.5 h, the mixture was cooled to RT and concentrated under
reduced pressure. The crude residue was purified by silica column
chromatography (10% to 30% EtOAc/Hex) to afford macrocycle 79-5.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.48ClN.sub.4O.sub.6: 667.33. found: 667.86.
[1066] Step 6. Preparation of 79-6. Macrocycle 79-5 (198 mg, 0.297
mmol) was treated with EtOAc (100 mL) and 5% Rh/alumina (100 mg).
H.sub.2 gas was bubbled through the solution for 1 min and the
reaction mixture was stirred at RT under an atmosphere of H.sub.2.
After 45 min, more 5% Rh/alumina (200 mg) was added. Again, H.sub.2
gas was bubbled through the solution for 1 min and the reaction
mixture was stirred at RT under an atmosphere of H.sub.2. After
another 1 h, the reaction mixture was filtered over celite and
concentrated under reduced pressure. The material (79-6) was
carried on without purification. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.36H.sub.50ClN.sub.4O.sub.6: 669.34. found: 669.63.
[1067] Step 7. Preparation of 79-7. Macrocycle 79-6 (0.297 mmol
theoretical) was treated with DCM (10 mL) and TFA (10 mL). The
reaction mixture was stirred at RT for 14 h, then concentrated
under reduced pressure. The crude residue was dissolved in EtOAc
and the organic solution was washed with saturated aqueous
NaHCO.sub.3 and 1 M citric acid. Brine was added after the citric
acid wash to break up the emulsion that formed. The organic layer
was dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The crude residue was purified by silica column
chromatography (100% EtOAc) to afford impure 79-7 that was carried
on without further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.32H.sub.42ClN.sub.4O.sub.6: 613.28. found: 613.22.
[1068] Step 8. Preparation of Example 79. Carboxcylic acid 79-7
(0.264 mmol theoretical) was treated with intermediate A9 (156 mg,
0.537 mmol), TBTU (170 mg, 0.528 mmol), DMAP (65 mg, 0.528 mmol),
DCM (2 mL) and DIPEA (0.23 mL, 1.3 mmol). The reaction mixture was
stirred at RT for 19 h then concentrated under reduced pressure.
The crude residue was purified by reverse phase HPLC to afford
Example 79 as a TFA salt. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.52ClF.sub.2N.sub.6O.sub.8S: 849.32. found: 849.16.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.17 (s, 1H), 7.86 (t,
J=8.1 Hz, 1H), 7.77 (t, J=3.5 Hz, 1H), 7.55 (dd, J=8.8, 2.3 Hz,
1H), 5.84 (td, J=55.7, 6.7 Hz, 1H), 5.62 (d, J=3.5 Hz, 1H), 4.98
(t, J=10.6 Hz, 1H), 4.53 (t, J=9.3 Hz, 1H), 4.42-4.26 (m, 2H), 4.19
(dd, J=12.0, 3.9 Hz, 1H), 3.34 (d, J=7.6 Hz, 1H), 2.99 (tt, J=8.2,
4.8 Hz, 2H), 2.78 (ddt, J=21.6, 14.2, 5.7 Hz, 2H), 2.28-2.12 (m,
1H), 2.08-1.16 (m, 19H), 1.16-0.96 (m, 17H), 0.58 (dd, J=8.3, 4.1
Hz, 1H), 0.55-0.44 (m, 1H).
Example 80
Preparation of
(3aR,7S,10S,11S,12R)-1-acetyl-7-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)--
1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-16-methoxy-11-met-
hyl-5,8-dioxo-1,2,3,3a,5,6,7,8,11,12,20,21,22,23,24,24a-hexadecahydro-1
OH-9,12-methanopyrrolo[2',3':
18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-10-carboxam-
ide
##STR00344## ##STR00345## ##STR00346##
[1070] Step 1. Preparation of 80-1: Amine 18-2 (195 mg, 0.495 mmol)
and Intermediate D18 (192.8 mg, 0.544 mmol) were dissolved in DMF
(10 mL). DIPEA (430 .mu.L, 2.48 mmol) was added followed by HATU
(207 mg, 0.544 mmol) at room temperature. After 1.5 h, the reaction
mixture was concentrated in vacuo and the crude residue was
directly purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford 80-1 (2:1 diastereomeric ratio
favoring desired). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.53ClN.sub.5O.sub.8: 730.3. found: 730.48.
[1071] Step 2. Preparation of 80-2: A stirred heterogeneous mixture
of 80-1 (314 mg, 0.431 mmol), PdCl.sub.2(dppf)*CH.sub.2Cl.sub.2
(35.2 mg, 0.043 mmol) and potassium vinyltrifluoroborate (86.6 mg,
0.646 mmol) in EtOH (2.2 mL) was sparged with argon for 15 min.
Triethylamine (320 .mu.L, 2.3 mmol) was added and the mixture was
heated to 80.degree. C. After 40 min, the reaction mixture was
cooled to ambient temperature and was diluted with toluene (5 mL).
The resulting mixture was concentrated and the crude residue was
directly purified by silica gel chromatography (0-100% ethyl
acetate/hexanes gradient) to afford 80-2 (2:1 diastereomeric ratio
favoring desired). LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.56N.sub.5O.sub.8: 722.4. found: 722.54.
[1072] Step 3. Preparation of 80-3: 80-2 (228 mg, 0.320 mmol) was
dissolved in DCE (64 mL) and the solution was sparged with Ar for
15 min. Zhan 1B catalyst (23 mg, 0.032 mmol) was added and the
resulting solution was stirred at 100.degree. C. under Ar. After 45
min, the reaction mixture was cooled to rt, was concentrated in
vacuo and was directly purified by silica gel chromatography
(0-100% ethyl acetate/hexanes gradient) to afford 80-3 (5:2
diastereomeric ratio favoring desired). LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.37H.sub.52N.sub.5O.sub.8: 694.37. found:
694.53.
[1073] Step 4: Preparation of 80-4: Olefin 80-3 (164 mg, 0.237
mmol) was dissolved in ethanol (1.19 mL) and the reaction vessel
was purged with Ar. Pd/C (10 wt % Pd, 25 mg) was added in a single
portion and the reaction vessel was purged thrice with H.sub.2. The
reaction was stirred at rt under 1 atm H.sub.2 for 2 h and was
diluted with ethyl acetate (10 mL). The resulting mixture was
filtered through a pad of Celite and concentrated to afford a crude
residue of 80-4 (5:2 diastereomeric ratio favoring desired) that
was used without further purification (LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.37H.sub.54N.sub.5O.sub.8: 696.39. found:
696.56.
[1074] Step 5. Preparation of 80-5: To a solution of 80-4 (164 mg,
240 .mu.mol) in DCM (1.2 mL) was added TFA (0.45 mL) at rt. After 7
h, the reaction mixture was diluted with ethyl acetate (50 mL) and
the resulting mixture was extracted with 1N aqueous sodium
hydroxide solution (40 mL). The aqueous layer was then slowly
acidified to pH=3 with concentrated hydrochloric acid, and was
extracted with ethyl acetate (2.times.50 mL). The combined organic
extracts were dried over anhydrous sodium sulfate and were
concentrated in vacuo. The residue was azeotropically dried with
toluene (3.times.5 mL) to afford 80-5 (5:2 diastereomeric ratio
favoring desired) that was used without further purification.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.33H.sub.46N.sub.5O.sub.8: 640.33. found: 640.48.
[1075] Step 6. Preparation of Example 80: To a solution of 80-5
(140 mg, 219 .mu.mol) and Intermediate A10 (133 mg, 438 .mu.mol) in
MeCN (1.1 mL) was added HATU (169 mg, 438 .mu.mol) followed by
DIPEA (190 .mu.L, 1.09 mmol) at rt under an argon atmosphere. After
15 h, the reaction mixture was concentrated in vacuo, was purified
by preparatory HPLC (Gemini 5u C18 110 .ANG. column, 5-100%
MeCN/H.sub.2O, 0.1% trifluoroacetic acid modifier) and was
lyophilized to afford Example 80 (5:2 diastereomeric ratio favoring
desired) as a light yellow solid TFA salt. Analytic HPLC RetTime:
7.91 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.58F.sub.2N.sub.7O.sub.10S: 890.39. found: 890.64.
.sup.1H NMR (400 MHz, CD.sub.3OD, Minor diastereomer denoted by *)
.delta. 9.18 (s, 1H), 9.14 (s, 1H*), 7.78 (br d, J=9.0 Hz, 1H,
1H*), 7.21 (br d, J=9.0 Hz, 1H, 1H*), 7.18 (br s, 1H, 1H*), 5.80
(br td, J.sub.H-F=55.8 Hz, J=6.8 Hz, 1H, 1H*), 5.64 (br s, 1H,
1H*), 5.23 (d, J=4.7 Hz, 1H*), 5.15 (d, J=4.7 Hz, 1H), 4.56 (d,
J=6.7 Hz, 1H, 1H*), 4.46 (d, J=12.1 Hz, 1H*), 4.41 (d, J=12.0 Hz,
1H), 4.30-4.22 (m, 1H, 1H*), 4.22-4.07 (m, 1H, 1H*), 4.02-3.79 (m,
1H, 1H*) 3.92 (br s, 3H, 3H*), 3.73-3.52 (m, 2H, 2H*), 3.05-2.68
(m, 3H, 3H*), 2.40-2.21 (m, 1H, 1H*), 2.13-1.94 (m, 4H, 4H*), 1.83
(s, 2H, 2H*), 1.75-1.20 (m, 12H, 12H*), 1.12 (s, 9H*),1.10 (s, 9H),
1.06 (br d, J=7.3 Hz, 3H, 3H*), 0.92-0.85 (m, 4H, 4H*).
Example 81
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-2-(difluorometh-
yl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-19,19-difluor-
o-15-methoxy-10-methyl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,-
23b-hexadecahydro-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,1-
9][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00347##
[1077] Example 81 was prepared in a similar fashion to Example 62,
substituting Intermediate A10 for Intermediate A9 in Step 5.
Example 81 was isolated. Analytic HPLC RetTime: 9.36 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.55F.sub.4N.sub.6O.sub.9S: 895.36. found: 895.59.
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 9.23 (s, 1H), 7.93 (d,
J=8.8 Hz, 1H), 7.31 (dd, J=8.8, 2.4 Hz, 1H), 7.26 (d, J=2.4 Hz,
1H), 5.80 (td, J.sub.H-F=56 Hz, J=6.8 Hz, 1H), 5.73 (d, J=3.2 Hz,
1H), 4.94 (d, J=7.2 Hz, 1H), 4.56 (d, J=6.8 Hz, 1H), 4.36 (d, J=6.8
Hz, 1H), 4.32 (s, 1H), 4.22-4.16 (dd, J=12, 4 Hz, 1H), 3.97 (s,
3H), 2.79-2.71 (m, 1H), 2.61-2.52 (m, 1H), 2.26-2.16 (m, 1H),
2.08-1.92 (m, 4H), 1.82-1.64 (m, 3H), 1.60-1.54 (m, 3H), 1.53-1.46
(m, 1H), 1.52 (s, 3H), 1.44-1.26 (m, 5H), 1.08 (s, 9H), 1.07-0.98
(m, 4H), 0.94-0.84 (m, 3H), 0.60-0.48 (m, 2H).
Example 82
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-2-(difluorometh-
yl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-19,19-difluor-
o-10-methyl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadec-
ahydro-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6-
]dioxadiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00348##
[1079] Intermediate 82-1 was prepared in a similar fashion to
intermediate 46-2, substituting intermediate E3 with E4 in Step 1.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.26H.sub.34F.sub.2N.sub.3O.sub.5: 506.25. found: 506.59.
[1080] Example 82 was prepared in a similar fashion to Example 62,
substituting Intermediate 82-1 for Intermediate 46-2 in Step 1.
Example 82 was isolated. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd
for C.sub.41H.sub.52F.sub.4N.sub.6O.sub.8S: 864.35. found: 865.43.
.sup.1H NMR (400 MHz, cdcl.sub.3) .delta. 9.82 (s, 1H), 7.89-7.72
(m, 2H), 7.67 (t, J=7.6 Hz, 1H), 6.93 (s, 1H), 6.12-5.65 (m, 2H),
5.34 (d, J=8.6 Hz, 1H), 4.90 (d, J=7.4 Hz, 1H), 4.45 (t, J=9.3 Hz,
2H), 4.27 (d, J=7.9 Hz, 1H), 4.13 (dd, J=11.9, 3.9 Hz, 1H),
2.77-2.64 (m, 2H), 2.27-2.12 (m, 1H), 2.13-1.86 (m, 4H), 1.82-1.19
(m, 15H), 1.18-0.98 (m, 13H), 0.89-0.77 (m, 2H), 0.53 (dd, J=13.3,
8.1 Hz, 1H), 0.43 (d, J=4.2 Hz, 1H).
Example 83
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18,18-difluoro--
3,6-dioxo-14-(trifluoromethoxy)-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetra-
decahydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadec-
ino[11,12-b]quinoxaline-8-carboxamide
##STR00349## ##STR00350## ##STR00351##
[1082] Step 1. Preparation of 83-1: HATU (3.06 g, 8.05 mmol) was
added slowly to a solution of 3,3-difluoro-2-oxopent-4-enoic acid
(1.03 g, 6.86 mmol) in 10 mL of DMF. A mixture of
4-(trifluoromethoxy)benzene-1,2-diamine (1.29 g, 6.71 mmol) and
DIPEA (1.4 mL, 8.05 mmol) in 12 mL of DMF was then added. After
stirring overnight, reaction mixture was poured into 175 mL of
water and extracted with ethyl acetate (4.times.100 mL). Combined
organics were washed with 50% brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. Resulting solid was purified via silica gel column
chromatography (0-25% ethyl acetate in hexanes) to yield
intermediate 83-1, the late eluting product. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.12H.sub.8F.sub.5N.sub.2O.sub.2: 307.04.
found: 307.29.
[1083] Step 2. Preparation of 83-2: A solution of 83-1 (924 mg,
3.01 mmol) in 2 mL DMF was treated with POCl.sub.3 (0.56 mL, 6.04
mmol) and heated at 80.degree. C. for 2.5 hours. After cooling to
room temperature, reaction mixture was diluted with 25 mL of EtOAc
and added slowly to 20 mL of water with vigorous stirring. Layers
were separated and aqueous was extracted with ethyl acetate.
Combined organics were washed subsequently with saturated aqueous
sodium bicarbonate and brine, dried over anhydrous sodium sulfate
and concentrated under reduced pressure to give intermediate 83-2.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.12H.sub.7ClF.sub.5N.sub.2O: 324.01. found: 324.13.
[1084] Step 3. Preparation of 83-3: Cs.sub.2CO.sub.3 (606 mg, 1.86
mmol) was added to a mixture of intermediate 83-2 (460 mg, 1.54
mmol) and intermediate B4 (564 mg, 1.79 mmol) in 12 mL of DMF at
room temperature. Reaction mixture was heated at 85.degree. C.
overnight. After cooling to room temperature, mixture was poured
into 50 mL of water and extracted with ethyl acetate (4.times.40
mL). Combined organics were washed with 90 mL of 50% brine, dried
over anhydrous sodium sulfate and concentrated under reduced
pressure. Resulting solid was purified via silica gel column
chromatography (0-30% ethyl acetate in hexanes) to give 83-3.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.28H.sub.35F.sub.5N.sub.3O.sub.6: 604.24. found: 604.20.
[1085] Step 4. Preparation of 83-4: Quinoxaline ether 83-3 (290 mg,
0.647 mmol) was dissolved in 4.1 mL of tert-butyl acetate and 1.1
mL of dichloromethane at room temperature. MeSO.sub.3H (0.25 mL,
3.88 mmol) was added dropwise and reaction mixture stirred at rt
for 2 h. The reaction mixture was transferred to a stirred mixture
of EtOAc (20 mL) and saturated aqueous NaHCO.sub.3 (30 mL). The
phases were separated, and the aqueous phase was extracted with
EtOAc (2.times.20 mL). The combined organic phase was dried over
anhydrous Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to
afford amine 83-4. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.23H.sub.27F.sub.5N.sub.3O.sub.4: 504.18. found: 504.31.
[1086] Step 5. Preparation of 83-5: HATU (260 mg, 0.684 mmol,
Oakwood) and DIPEA (0.40 mL, 2.30 mmol) were added to a mixture of
83-4 (258 mg, 0.512 mmol) and Intermediate D11 (177 mg, 0.657 mmol)
in 7 mL of acetonitrile under argon. After stirring overnight, the
reaction mixture was concentrated under reduced pressure and the
resulting residue was purified by silica gel chromatography (0-20%
ethyl acetate in hexanes) to yield 83-5. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.37H.sub.48F.sub.5N.sub.4O.sub.7:
755.34. found: 755.49.
[1087] Step 6. Preparation of 83-6: A mixture of 83-5 (215 mg,
0.285 mmol) and Zhan 1B catalyst (29 mg, 0.040 mmol, Strem) in 60
mL of DCE was deoxygenated with argon for 15 minutes. The mixture
was then heated at reflux for 90 minutes. After cooling to room
temperature, reaction mixture was concentrated in vacuo. The
resulting residue was purified by silica gel chromatography (0-40%
ethyl acetate in hexanes) to yield 83-6. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.35H.sub.44F.sub.5N.sub.4O.sub.7:
727.31. found: 727.43.
[1088] Step 7. Preparation of 83-7: Palladium on carbon (10 wt %
Pd, 40 mg, 0.038 mmol) was added to a solution of 83-6 (129 mg,
0.178 mmol) in 9 mL of ethanol. The atmosphere was replaced with
hydrogen and the reaction stirred overnight. The reaction mixture
was filtered over Celite and washed with ethanol. Filtrate was
concentrated in vacuo to yield a residue, which was purified via
silica gel column chromatography (0-30% ethyl acetate in hexanes)
to yield 83-7. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.35H.sub.46F.sub.5N.sub.4O.sub.7: 729.32. found: 729.45.
[1089] Step 8. Preparation of 83-8: TFA (0.62 mL, 8.09 mmol) was
added slowly to a solution of 83-7 (79 mg, 0.109 mmol) in 1.8 mL of
dichloromethane. After 4 hours, mixture was concentrated under
reduced pressure to near dryness. Resulting residue was taken up in
10 mL of ethyl acetate, washed with 8 mL of water, 8 mL of sat.
NaHCO.sub.3 (aq), and separated. Aqueous layers were extracted with
ethyl acetate (3.times.10 mL). Combined organics were washed with
10 mL of brine, dried over anhydrous Na.sub.2SO.sub.4, filtered,
and concentrated in vacuo to yield 83-8, which was used in the next
step without further purification. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.31H.sub.38F.sub.5N.sub.4O.sub.7: 673.26. found:
673.10.
[1090] Step 9. Preparation of Example 83: HATU (84 mg, 0.221 mmol,
Oakwood) and DIPEA (0.095 mL, 0.547 mmol) were added to a mixture
of 83-8 (72 mg, 0.107 mmol) and Intermediate A10 (66 mg, 0.217
mmol) in 4 mL of acetonitrile under argon. After stirring for
overnight, reaction mixture was taken up in 20 mL of ethyl acetate
and washed with 10 mL of 1 N aqueous HCl. The aqueous layer was
extracted three times with ethyl acetate. Combined organics were
washed with 50% brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The resulting residue was
purified by silica gel chromatography (0-50% ethyl acetate in
hexanes) and reverse phase prep HPLC (50-100% acetonitrile in
water, with 0.1% trifluoroacetic acid buffer) to yield the
trifluoroacetic acid salt of Example 83. Analytic HPLC RetTime:
9.12 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.50F.sub.7N.sub.6O.sub.9S: 923.32. found: 923.10.
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 9.26 (s, 1H), 8.01-7.91
(m, 2H), 7.78-7.63 (m, 1H), 5.95 (d, J=3.6 Hz, 1H), 5.83 (td,
J.sub.H-F=61 Hz, J=6.0 Hz, 1H), 4.59 (d, J=7.2 Hz, 1H), 4.42 (d,
J=12.4 Hz, 1H), 4.35 (s, 1H), 4.22-4.11 (m, 1H), 3.72-3.66 (m, 1H),
2.71-2.49 (m, 2H), 2.18-1.94 (m, 3H), 1.90-1.75 (m, 3H), 1.74-1.62
(m, 2H), 1.60-1.48 (m, 3H), 1.51 (s, 3H), 1.50-1.24 (m, 4H),
1.22-1.18 (m, 2H), 1.08 (s, 9H), 1.07-0.84 (m, 5H), 0.81-0.64 (m,
1H), 0.54-0.44 (m, 1H).
Example 84
Preparation of
(1aR,5S,8S,9S,10R,19E,22aR)-5-tert-butyl-14-cyano-N-[(1R,2R)-2-(difluorom-
ethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18-
,18-difluoro-3,6-dioxo-1,1a,3,4,5,6,9,10,18,21,22,22a-dodecahydro-8H-7,10--
methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinox-
aline-8-carboxamide
##STR00352##
[1092] Step 1: Preparation of Example 84. Crude Example 78 (8.7 mg,
0.01 mmol) was redissolved in ACN (0.3 mL) and treated with DDQ
(3.4 mg, 0.015 mmol). After 10 min, the solution was directly
purified by reverse phase HPLC (Gemini 5u C18 110 .ANG. column,
50-100% ACN/H.sub.2O+0.1% TFA) and lyophilized to afford the TFA
salt of Example 84. Analytical HPLC RetTime: 8.385 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.47F.sub.4N.sub.7O.sub.8S: 861.90. found: 862.89.
.sup.1H NMR (400 MHz, CD3OD).sup.1H NMR (400 MHz, CD3OD) .delta.
9.21 (s, 1H), 8.25 (d, 1H), 8.20 (d, 1H), 0.7.91 (dd, 1H), 6.32 (m,
2H), 5.97-5.61 (m, 2H), 4.82 (m, 1H), 4.58-4.13 (m, 4H), 3.71-3.49
(m, 3H), 2.61 (m, 2H), 2.23 (m, 1H), 2.00-1.80 (m, 3H), 1.56-1.20
(m, 10H), 1.20 (m, 3H), 1.07 (m, 8H), 0.98-0.82 (m, 3H)., 0.55 (m,
1H).
Example 85
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-14-(difluoromethoxy)-N-[(1R,2R)-2-(d-
ifluoromethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9--
ethyl-18,18-difluoro-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetrad-
ecahydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadeci-
no[11,12-b]quinoxaline-8-carboxamide
##STR00353##
[1094] Example 85 was prepared similarly to Example 83, by using
intermediate E7 in place of 83-2 in step 3. Analytical HPLC
RetTime: 8.725 min. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.50F.sub.6N.sub.6O.sub.9S: 904.92. found: 905.16.
.sup.1H NMR (400 MHz, CD3OD) .delta. 9.23 (s, 1H), 7.88 (d, 1H),
7.76 (d, 1H), 0.7.62 (dd, 1H), 7.03 (dd, 1H), 5.94-5.65 (m, 3H),
4.57-4.14 (m, 4H), 3.66 (m, 1H), 2.57 (m, 2H), 2.01-1.97 (m, 3H),
1.82-1.77 (m, 3H), 1.64 (m, 1H), 1.57-1.33 (m, 10H), 1.20 (m, 3H),
1.06-0.87 (m, 12H), 0.87 (m, 2H)., 0.48 (m, 1H).
Example 86
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-N-[(1R,2R)-2-(difluorometh-
yl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-15-fluoro-10--
methyl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadecahydr-
o-1H,9H-8,11-methanocyclopropa[4',5']cyclopenta[1',2':18,19][1,10,3,6]diox-
adiazacyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00354## ##STR00355## ##STR00356##
[1096] Step 1. Preparation of 86-1: To a solution of E8 (1.5 g,
5.75 mmol) and B1 (1.9 g, 6.34 mmol) in MeCN (50 mL) is added
Cs.sub.2CO.sub.3 (3.09 g, 9.49 mmol). After stirring at rt for 60
h, the reaction mixture was filtered over celite and concentrated.
The crude residue was purified by silica gel chromatography (5-35%
EtOAc/hexanes) to yield product 86-1. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.23H.sub.30ClFN.sub.3O.sub.5-- Boc:
482.13. found: 382.04.
[1097] Step 2. Preparation of 86-2: To a solution of 86-1 (747 mg,
1.55 mmol) in CH.sub.2Cl.sub.2 (5 mL) is added HCl (5 mL, 4 M in
dioxane) and allowed to stir for 3 h. The reaction mixture was
concentrated to give a residue that was used subsequently without
further purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.18H.sub.23Cl.sub.2FN.sub.3O.sub.3--HCl: 382.13. found:
382.08.
[1098] Step 3. Preparation of 86-3: To a solution of 86-2 (397 mg,
0.95 mmol), D12 (308 mg, 0.95 mmol) and BEP (312 mg, 1.14 mmol) in
EtOAc (9 mL) and NMP (1 mL) was added DIPEA (0.7 mL, 3.8 mmol) and
the reaction was stirred at 50.degree. C. overnight. The reaction
was quenched with sat. NaHCO.sub.3 solution and extracted with
EtOAc, washed subsequently with brine, dried over magnesium sulfate
and concentrated. The crude product was purified by silica gel to
yield 86-3. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.49ClFN.sub.4O.sub.6: 687.33. found: 687.44.
[1099] Step 4. Preparation of 86-4: To a solution of 86-3 (266 mg,
0.39 mmol), TEA (0.08 mL, 0.58 mmol) and potassium
vinyltrifluoroborate (78 mg, 0.58 mmol) in EtOH (8 mL) was added
PdCl.sub.2(dppf) (32 mg, 0.04 mmol). The reaction was degassed with
N.sub.2 for 10 min and heated to 75.degree. C. for 1 h. The
reaction was quenched with sat. NaHCO.sub.3 solution and extracted
with EtOAc, washed subsequently with brine, dried over magnesium
sulfate and concentrated. The residue was purified using silica gel
chromatography (0-25% EtOAc/hexanes) to give 86-4. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.38H.sub.52FN.sub.4O.sub.6: 679.39.
found: 679.52.
[1100] Step 5 and 6. Preparation of 86-5: To a solution of 86-4
(262 mg, 0.38 mmol) in DCE (50 mL) was added Zhan 1B catalyst (28
mg, 0.04 mmol) and the reaction was degassed for 25 minutes with
N.sub.2. The reaction was heated to 100.degree. C. for 1 h, allowed
to cool to rt and concentrated. The crude product was purified by
silica gel chromatography (0-30% EtOAc/hexanes) to give olefin
product (182 mg; LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.48FN.sub.4O.sub.6: 651.36. found: 651.38) that was
taken up in EtOH (5 mL) and EtOAc (1 mL) and treated with Pd/C
(10%, 55 mg). The atmosphere was replaced with hydrogen and stirred
at rt for 1.25 h. The reaction was filtered over Celite, washed
with EtOAc and concentrated to give 86-5 that was used subsequently
without further purification. LCMS-ESI.sup.+ (m/z): [M+H].sup.+
calcd for C.sub.36H.sub.50FN.sub.4O.sub.6: 653.37. found:
653.46.
[1101] Step 7. Preparation of 86-6: To a solution of 86-5 (182 mg,
0.28 mmol) in DCM (3 mL) was added TFA (3 mL) and stirred at rt for
18 h. The reaction was diluted with EtOAc, washed with H.sub.2O,
basicified to pH 7 with sat. NaHCO.sub.3 solution, washed with 1M
citric acid solution, dried over magnesium sulfate, and
concentrated to give a residue of 86-6 that was used subsequently
without further purification. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.32H.sub.42FN.sub.4O.sub.6: 597.31. found: 597.15.
[1102] Step 8. Preparation of Example 86: To a solution of 86-6 (24
mg, 0.04 mmol), intermediate A10 (18 mg, 0.06 mmol), TBTU (23 mg,
0.07 mmol) and DMAP (7 mg, 0.06 mmol) in DMF (3 mL) was added DIPEA
(35 .mu.L, 0.20 mmol) and the reaction was stirred at rt for 3 h.
Additional intermediate A10 (18 mg, 0.06 mmol), TBTU (23 mg, 0.07
mmol), DMAP (7 mg, 0.06 mmol), and DIPEA (35 .mu.L, 0.20 mmol) was
added and the reaction was stirred at rt for 16 h. The crude
material was purified by reverse phase HPLC (Gemini, 30-85%
ACN/H.sub.2O+0.1% TFA) and lyophilized to give Example 86 as a TFA
salt. Analytical HPLC RetTime: 9.25 min. LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.41H.sub.54F.sub.3N.sub.6O.sub.8S:
847.37; found: 847.18. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
9.18 (s, 1H), 8.13-7.84 (m, 2H), 7.59-7.21 (m, 2H), 6.07-5.58 (m,
2H), 5.00 (d, J=7.4 Hz, 1H), 4.57 (d, J=7.0 Hz, 1H), 4.45-4.27 (m,
2H), 4.20 (dd, J=12.0, 4.0 Hz, 1H), 3.11-2.94 (m, 3H), 2.92-2.70
(m, 4H), 2.32-2.14 (m, 1H), 2.10-1.94 (m, 2H), 1.86 (m, 1H), 1.77
(d, J=14.5 Hz, 1H), 1.74-1.21 (m, 15H), 1.21-1.01 (m, 10H),
1.00-0.84 (m, 2H), 0.60 (m, 1H), 0.53 (m, 1H).
Example 87
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-14-cyano-N-[(1R,2R)-2-(difluoromethy-
l)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18,18--
difluoro-3,6-dioxo-1,1a,3,4,5,6,9,
10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1,1-
0,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00357##
[1104] Steps 1 and 2. Preparation of Example 87. To a solution of
Example 84 (100 mg, 0.11 mmol) in EtOAc (3 mL) was added Pd/C (10
wt % Pd, 30 mg). The reaction vessel was purged twice with H.sub.2
and was stirred at rt under 1 atm H.sub.2 for 6 h. After which time
the reaction mixture was filtered through a pad of celite and
concentrated. The reaction reduced the quinoxaline ring. The crude
material was redissolved in ACN (5 mL) and treated with DDQ (34 mg,
0.15 mmol). After 1 h, the solution was directly purified by
reverse phase HPLC (Gemini 5u C18 110 .ANG. column, 50-100%
ACN/H.sub.2O+0.1% TFA) and lyophilized to afford the TFA salt of
Example 87. Analytical HPLC RetTime: 8.463 min. LCMS-ESI.sup.+
(m/z): [M+H].sup.+calcd for C.sub.40H.sub.49F.sub.4N.sub.7O.sub.8S:
863.92. found: 864.18. .sup.1H NMR (400 MHz, CD3OD) .delta. 9.24
(s, 1H), 8.27 (d, 1H), 8.20 (d, 1H), 0.7.91 (dd, 1H), 5.93-5.82 (m,
3H), 4.88 (m, 1H), 4.58-4.13 (m, 5H), 3.71-3.49 (m, 3H), 2.59 (m,
2H), 2.03-1.96 (m, 3H), 1.82-1.77 (m, 3H), 1.65-1.35 (m, 11H), 1.20
(m, 3H), 1.06-0.87 (m, 8H), 0.71 (m, 2H)., 0.48 (m, 1H).
Example 88
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-14-chloro-N-[(1R,2R)-2-(difluorometh-
yl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-18,18-difluor-
o-9-methyl-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8-
H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b-
]quinoxaline-8-carboxamide
##STR00358## ##STR00359## ##STR00360##
[1106] Step 1. Preparation of 88-1: HATU (4.56 g, 12 mmol) was
added slowly to a solution of 3,3-difluoro-2-oxopent-4-enoic acid
(1.52 g, 10.1 mmol) in 14 mL of DMF. A mixture of
4-chlorobenzene-1,2-diamine (1.43 g, 10 mmol) and DIPEA (2.1 mL, 12
mmol) in 20 mL of DMF was then added. After stirring overnight,
reaction mixture was poured into 30 mL of 1 N aqueous HCl and
extracted with ethyl acetate (5.times.40 mL). Combined organics
were dried over anhydrous Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. Resulting solid was purified
via silica gel column chromatography (0-45% ethyl acetate in
hexanes) to yield intermediate 88-1 as the late eluting product.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 12.1 (s, 1H), 7.99 (m,
1H), 7.61-7.58 (m, 1H), 7.33-7.31 (m, 1H), 6.61-6.48 (m, 1H),
5.96-5.90 (m, 1H), 5.67-5.63 (m, 1H).
[1107] Step 2. Preparation of 88-2: A solution of intermediate 88-1
(648 mg, 2.53 mmol) in 2 mL DMF was treated with POCl.sub.3 (0.49
mL, 5.26 mmol) and heated at 80.degree. C. for 3 hours. After
cooling to room temperature, reaction mixture was diluted with 20
mL of EtOAc and added slowly to 15 mL of water with vigorous
stirring. Layers were separated and aqueous was extracted with
ethyl acetate. Combined organics were washed subsequently with
saturated aqueous sodium bicarbonate and brine, dried over
anhydrous sodium sulfate and concentrated under reduced pressure to
give intermediate 88-2. .sup.1H NMR (400 MHz, CDCl.sub.3) b 8.184
(d, J=1.6 Hz, 1H), 8.01 (d, J=8.8 Hz, 1H), 7.82 (dd, J=9.4, 2 Hz,
1H), 6.56-6.43 (m, 1H), 5.88 (m, 1H), 5.70 (d, J=10.8 Hz, 1H).
[1108] Step 3. Preparation of 88-3: Cs.sub.2CO.sub.3 (660 mg, 2.03
mmol) was added to a mixture of intermediate 88-2 (425 mg, 1.54
mmol) and intermediate B1 (570 mg, 1.89 mmol) in 9 mL of DMF at
room temperature. Reaction mixture was heated at 85.degree. C.
overnight. After cooling to room temperature, mixture was poured
into 40 mL of water and extracted with ethyl acetate (4.times.30
mL). Combined organics were washed with 75 mL of 50% brine, dried
over anhydrous sodium sulfate and concentrated under reduced
pressure. Resulting solid was purified via silica gel column
chromatography (0-20% ethyl acetate in hexanes) to give 88-3.
LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.26H.sub.33ClF.sub.2N.sub.3O.sub.5: 540.20. found: 540.12.
[1109] Step 4. Preparation of 88-4: Quinoxaline ether 88-3 (458 mg,
0.848 mmol) was dissolved in 4.2 mL of tert-butyl acetate and 1.2
mL of dichloromethane at room temperature. MeSO.sub.3H (0.30 mL,
4.67 mmol) was added dropwise and reaction mixture stirred at rt
for 2 h. The reaction mixture was transferred to a stirred mixture
of EtOAc (20 mL) and saturated aqueous NaHCO.sub.3 (30 mL). The
phases were separated, and the aqueous phase was extracted with
EtOAc (2.times.20 mL). The combined organic phase was dried over
anhydrous Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to
afford amine 88-4 as a yellow solid film LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.21H.sub.25ClF.sub.2N.sub.3O.sub.3:
440.15. found: 440.29.
[1110] Step 5. Preparation of 88-5: HATU (360 mg, 0.947 mmol,
Oakwood) and DIPEA (0.51 mL, 2.91 mmol) were added to a mixture of
88-4 (320 mg, 0.727 mmol) and Intermediate D11 (237 mg, 0.880 mmol)
in 10 mL of acetonitrile under argon. After stirring overnight, the
reaction mixture was concentrated under reduced pressure and the
resulting residue was purified by silica gel chromatography (0-20%
ethyl acetate in hexanes) to yield 88-5. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.35H.sub.46ClF.sub.2N.sub.4O.sub.6:
691.30. found: 691.50.
[1111] Step 6. Preparation of 88-6: A mixture of 88-5 (390 mg,
0.564 mmol) and Zhan 1B catalyst (55 mg, 0.075 mmol, Strem) in 100
mL of DCE was deoxygenated with argon for 15 minutes. The mixture
was then heated at reflux for 110 minutes. After cooling to room
temperature, reaction mixture was concentrated in vacuo. The
resulting residue was purified by silica gel chromatography (0-25%
ethyl acetate in hexanes) to yield 88-6. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.33H.sub.42ClF.sub.2N.sub.4O.sub.6:
663.27. found: 663.33.
[1112] Step 7. Preparation of mixture of 88-7: Rhodium on alumina
(5 wt % Rh, 31 mg, 0.015 mmol) was added to a solution of 88-6 (90
mg, 0.136 mmol) in 9 mL of ethanol. The atmosphere was replaced
with hydrogen and mixture was stirred overnight. The reaction was
filtered over Celite, washing with ethanol. LC/MS analysis
indicated about 60% starting material remained. A solution of the
residue in 8 mL of ethanol was resubjected to hydrogenation
conditions utilizing 25 mg of Rhodium on alumina (5 wt % Rh)
overnight. The reaction was filtered over Celite, washing with
ethanol. Filtrate was concentrated in vacuo to yield as residue,
which was purified via silica gel column chromatography (0-30%
ethyl acetate in hexanes) to yield 88-7. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.33H.sub.44ClF.sub.2N.sub.4O.sub.6:
665.28. found: 665.48.
[1113] Step 8. Preparation of 88-8: TFA (0.45 mL, 5.86 mmol) was
added slowly to a solution of 88-7 (52 mg, 0.078 mmol) in 2 mL of
dichloromethane. After 3 hours, mixture was concentrated under
reduced pressure to near dryness. Resulting residue was taken up in
10 mL of ethyl acetate, washed with 8 mL of water, 8 mL of sat.
NaHCO.sub.3 (aq), and separated. Aqueous layers were extracted with
ethyl acetate (3.times.10 mL). Combined organics were washed with
10 mL of brine, dried over anhydrous MgSO.sub.4, filtered, and
concentrated in vacuo to yield 88-8, which was used in the next
step without further purification. LCMS-ESI (m/z): [M+H].sup.+
calcd for C.sub.29H.sub.36ClF.sub.2N.sub.4O.sub.6: 609.22. found:
609.42.
[1114] Step 9. Preparation of Example 88: HATU (58 mg, 0.153 mmol,
Oakwood) and DIPEA (0.065 mL, 0.374 mmol) were added to a mixture
of 88-8 (45 mg, 0.074 mmol) and Intermediate A10 (49 mg, 0.161
mmol) in 2.5 mL of acetonitrile under argon. After stirring for
overnight, reaction mixture was taken up in 15 mL of ethyl acetate
and washed with 10 mL of 1 N aqueous HCl. The aqueous layer was
extracted three times with ethyl acetate. Combined organics were
washed with 50% brine, dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. The resulting residue was
purified by silica gel chromatography (0-50% ethyl acetate in
hexanes) and reverse phase prep HPLC (50-100% acetonitrile in
water, with 0.1% trifluoroacetic acid buffer) to yield the
trifluoroacetic acid salt of Example 88. Analytic HPLC RetTime:
8.92 min. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.38H.sub.48ClF.sub.4N.sub.6O.sub.8S: 859.28. found: 859.42.
.sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 9.23 (s, 1H), 8.10 (s,
1H), 7.90 (d, J=8.8 Hz, 1H), 7.81 (d, J=8.8 Hz, 1H), 5.81 (td,
J.sub.H-F=56 Hz, J=6.0 Hz, 1H), 5.69-5.66 (m, 1H), 4.56 (d, J=7.2
Hz, 1H), 4.43 (d, J=12 Hz, 1H), 4.34 (s, 1H), 4.22-4.16 (dd, J=12,
4 Hz, 1H), 3.71-3.66 (m, 1H), 2.83-2.76 (m, 1H), 2.61-2.48 (m, 1H),
2.11-1.94 (m, 4H), 1.88-1.72 (m, 4H), 1.71-1.62 (m, 1H), 1.58-1.54
(m, 2H), 1.51 (s, 3H), 1.50-1.36 (m, 2H), 1.09 (s, 9H), 1.08-1.01
(m, 3H), 1.01-0.94 (m, 2H), 0.93-0.86 (m, 2H), 0.80-0.68 (m, 1H),
0.52-0.46 (m, 1H).
Example 89
Preparation of
(1aR,5S,8S,9S,10R,19E,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1--
{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18,18-diflu-
oro-14-methoxy-3,6-dioxo-1,1a,3,4,5,6,9,10,18,21,22,22a-dodecahydro-8H-7,1-
0-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quin-
oxaline-8-carboxamide
##STR00361##
[1116] Step 1. Preparation of 89-1: 17-4 (95 mg, 0.14 mmol) in 0.4
mL DCM was treated with 0.4 mL TFA and stirred at rt for 2 h. The
reaction mixture was diluted with 5 mL DCM and then treated with
water and saturated sodium bicarbonate to pH 6.5. The layers were
separated and the organic phase was washed once more with water,
then dried over anhydrous sodium sulfate, filtered and concentrated
to give 89-1. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.31H.sub.39F.sub.2N.sub.4O.sub.7: 617.3. found: 616.7.
[1117] Step 2. Preparation of Example 89: A mixture of 89-1 from
step 1 (41 mg, 0.066 mmol), Intermediate A10 (24 mg, 0.079 mmol),
HATU (30 mg, 0.079 mmol), and DIPEA (0.057 mL, 0.33 mmol) in DMF
(0.4 mL) was stirred at rt overnight. The mixture was diluted with
2N HCl (1 mL) and extracted with dichloromethane. The organic phase
was dried over sodium sulfate, filtered and concentrated. The crude
product mixture was purified by reverse phase prep HPLC (10-99%
acetonitrile in water, with 0.1% trifluoroacetic acid buffer) to
give Example 89. Analytic HPLC RetTime: 8.65 min. LCMS-ESI (m/z):
[M+H].sup.+ calcd for C.sub.40H.sub.51F.sub.4N.sub.6O.sub.9S:
867.3. found: 866.9. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
9.890 (s, 1H), 7.98 (d, J=9.2 Hz, 1H), 7.28 (dd, J=8.8, 2.4 Hz,
1H), 7.06 (d, J=2.8 Hz, 1H), 6.75 (br s, 1H), 6.30-5.93 (m, 2H),
5.92 (td, J.sub.H-F=52 Hz, J=6.8 Hz, 1H), 5.47 (d, J=10 Hz, 1H),
4.53 (d, J=12 Hz, 1H), 4.48 (d, J=10.4 Hz, 1H), 4.42 (d, J=6.8 Hz,
1H), 4.07 (dd, J=11.6, 3.2 Hz, 1H), 3.98-3.94 (m, 1H), 3.95 (s,
3H), 3.57 (m, 1H), 2.60-2.48 (m, 2H), 2.20 (m, 1H), 2.06 (m, 1H),
1.90 (m, 1H), 1.80 (m, 1H), 1.63 (m, 2H), 1.50 (s, 3H), 1.56-1.36
(m, 2H), 1.26 (m, 1H), 1.19 (t, J=7.2 Hz, 3H), 1.09 (s, 9H),
1.03-0.93 (m, 2H), 0.85 (m, 2H), 0.76 (m, 1H), 0.53 (m, 1H).
Example 90
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(1R,2R)-2-(difluoromethyl)-1-{[(1-
-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-9-ethyl-18-fluoro-14-me-
thoxy-3,6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,1-
0-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quin-
oxaline-8-carboxamide
##STR00362##
[1119] Further purification of a synthesis of compound 17 by
reverse phase prep HPLC (60-88% acetonitrile in water, with 0.1%
trifluoroacetic acid buffer) allowed isolation of example 93 as a
minor side product. Analytic HPLC RetTime: 8.64 min. LCMS-ESI
(m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.54F.sub.3N.sub.6O.sub.9S: 851.4. found: 851.4.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.93 (br s, 1H), 7.88 (d,
J=9.1 Hz, 1H), 7.22 (d, J=2.4 Hz, 1H), 7.06 (d, J=2.4 Hz, 1H), 6.55
(s, 1H), 5.91 (td, J.sub.H-F=136 Hz, J=8 Hz, 1H), 5.81 (td,
J.sub.H-F=52 Hz, J=8 Hz, 1H), 5.30 (d, J=9.7 Hz, 1H), 4.44 (d,
J=12.0 Hz, 1H), 4.38 (d, J=6.7 Hz, 1H), 4.32 (d, J=9.8 Hz, 1H),
4.07 (m, 1H), 3.93 (s, 3H), 3.72 (m, 1H), 2.59 (m, 1H), 2.35 (m,
1H), 2.06 (m, 4H), 1.88 (m, 1H), 1.78 (m, 1H), 1.71-1.52 (m, 4H),
1.48 (s, 3H), 1.48-1.41 (m, 2H), 1.23 (m, 2H) 1.21 (t, J=8.0 Hz,
3H), 1.08 (s, 9H), 1.05-0.90 (m, 2H), 0.84 (m, 2H), 0.66 (m, 1H),
0.48 (m, 1H).
Example 91
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-{(1R,2S)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-ethenylcyclopropyl}-9-ethyl-14-methoxy-3,6-dioxo-1,1a,3,4,5,6,-
9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclopropa[18,19][1,-
10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carboxamide
##STR00363##
[1121] Example 91 was prepared similarly to Example 1 substituting
Intermediate A1 for Intermediate A10 in Step 8. The TFA salt of
Example 91 was isolated. Analytic HPLC RetTime: 8.72 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.55N.sub.6O.sub.9S: 795.96. found: 795.94. .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 9.03 (s, 1H); 7.80 (d, J=9.2 Hz, 1H);
7.24 (dd, J=9.2, 2.4 Hz, 1H); 7.16 (d, J=2.4 Hz, 1H); 5.90 (d,
J=3.6 Hz, 1H); 5.68 (m, 1H); 5.25 (d, J=17.2 Hz, 1.6 Hz, 1H); 5.10
(d, J=11.2, 1.6 Hz, 1H); 4.57 (d, J=6.8 Hz, 1H); 4.39 (br s, 1H);
4.37 (d, J=9.2 Hz, 1H); 4.16 (dd, J=12.8, 4.4 Hz, 1H); 3.93 (s,
3H); 3.77-3.72 (m, 1H); 3.02-2.88 (m, 1H): 2.86-2.75 (m, 1H);
2.64-2.54 (m, 1H); 2.18 (q, J=8.8 Hz, 1H): 1.90-1.66 (m, 4H);
1.66-1.40 (m, 6H); 1.38-1.32 (m, 1H); 1.30-1.20 (m, 5H); 1.10 (s,
9H); 1.14-1.02 (m, 2H); 0.77-0.68 (m, 1H); 0.54-0.45 (m, 1H).
Example 92
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-{(1R,2S)-1-[(cyclopropylsulfonyl)c-
arbamoyl]-2-ethenylcyclopropyl}-9-ethyl-18,18-difluoro-14-methoxy-3,6-diox-
o-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methanocyclo-
propa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline-8-carb-
oxamide
##STR00364##
[1123] Example 92 was prepared in a similar fashion to Example 17,
substituting Intermediate A1 for Intermediate A10 in Step 7.
Example 92 was isolated. Analytic HPLC RetTime: 8.75 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.40H.sub.53F.sub.2N.sub.6O.sub.9S: 831.36. found: 831.25.
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 9.98 (s, 1H), 7.96 (d,
J=9.2 Hz, 1H), 7.40-7.19 (m, 1H), 7.08 (s, 1H), 6.56 (s, 1H), 5.91
(d, J=3.8 Hz, 1H), 5.86-5.64 (m, 1H), 5.34 (d, J=9.7 Hz, 1H), 5.21
(d, J=17.2 Hz, 1H), 5.10 (d, J=10.3 Hz, 1H), 4.53-4.26 (m, 2H),
4.15-4.02 (m, 1H), 3.95 (s, 3H), 3.73-3.57 (m, 1H), 2.97-2.81 (m,
1H), 2.64-2.37 (m, 2H), 2.21-2.06 (m, 1H), 2.06-1.88 (m, 2H),
1.88-1.55 (m, 4H), 1.55-1.12 (m, 10H), 1.07 (s, 9H), 1.02-0.78 (m,
5H), 0.78-0.61 (m, 1H), 0.47 (q, J=7.3, 6.2 Hz, 1H).
Example 93
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-9-ethyl-N-[(2R)-2-ethyl-1-{[(1-methy-
lcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-18,18-difluoro-14-methoxy-3,6-
-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradeca
hydro-8H-7,10-methanocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino-
[11,12-b]quinoxaline-8-carboxamide
##STR00365##
[1125] Example 93 was prepared in a similar fashion to Example 17,
substituting Intermediate A4 for Intermediate A10 in Step 7.
Example 93 was isolated. Analytic HPLC RetTime: 8.03 min. LCMS-ESI
(m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.57F.sub.2N.sub.6O.sub.9S: 847.39. found: 846.99.
.sup.1H NMR (400 MHz, cdcl.sub.3) .delta. 7.95 (d, J=8.9 Hz, 1H),
7.27 (m, 1H), 7.08 (s, 1H), 6.65 (s, 1H), 5.91 (s, 1H), 5.41 (d,
J=9.0 Hz, 1H), 4.82 (m, 2H), 4.47 (d, J=6.2 Hz, 1H), 4.35 (dd,
J=35.7, 10.7 Hz, 2H), 4.07 (m, 1H), 3.94 (s, 3H), 3.63 (m, 1H),
2.50 (m, 2H), 1.95 (m, 2H), 1.94 (m, 2H), 1.78 (m, 3H), 1.64 (m,
4H), 1.48 (m, 6H), 1.19 (m, 4H), 1.07 (s, 9H), 1.05-0.88 (m, 4H),
0.88-0.75 (m, 1H), 0.67 (m, 1H), 0.47 (m, 1H).
Example 94
Preparation of
(1aR,5S,8S,9S,10R,22aR)-5-tert-butyl-N-[(2R)-1-[(cyclopropylsulfonyl)carb-
amoyl]-2-(difluoromethyl)cyclopropyl]-9-ethyl-18,18-difluoro-14-methoxy-3,-
6-dioxo-1,1a,3,4,5,6,9,10,18,19,20,21,22,22a-tetradecahydro-8H-7,10-methan-
ocyclopropa[18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxaline--
8-carboxamide
##STR00366##
[1127] Example 94 was prepared in a similar fashion to Example 17,
substituting Intermediate A9 for Intermediate A10 in Step 7.
Example 94 was isolated. Analytic HPLC RetTime: 8.71 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.39H.sub.51F.sub.4N.sub.6O.sub.9S: 855.34. found: 855.26.
.sup.1H NMR (400 MHz, CDCl.sub.3) (10.22 (s, 1H), 8.02 (d, J=9.2
Hz, 1H), 7.33 (d, J=12 Hz, 1H), 7.12 (s, 1H), 5.95 (td, J.sub.HF=52
Hz, J=8 Hz, 1H), 5.50 (d, J=9.7 Hz, 1H), 4.53 (d, J=6.4 Hz, 1H),
4.46 (dd, J=26.4, 10.7 Hz, 2H), 4.13 (d, J=11.5 Hz, 1H), 4.00 (s,
3H), 3.68 (m, 1H), 2.91 (m, 1H), 2.57 (m, 3H), 2.13 (m, 2H), 1.94
(m, 2H), 1.73 (m, 3H), 1.50 (m, 3H), 1.33 (m, 3H), 1.22 (t, J=7.2
Hz, 3H), 1.13 (s, 9H), 1.00-0.95 (m, 4H), 0.95-0.85 (m, 1H), 0.69
(m, 1H), 0.51 (m, 1H).
Example 95
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-15-cyano-N-[(1R,2R)-2-(dif-
luoromethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-19,1-
9-difluoro-10-methyl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,
19,20,21,22,23,23a,23b-hexadecahydro-1H,9H-8,11-methanocyclopropa[4',5']c-
yclopenta[1',2':18,19][1,10,3,6]dioxadiazacyclononadecino[11,12-b]quinoxal-
ine-9-carboxamide
##STR00367##
[1129] Intermediate 95-1 was prepared in a similar fashion to
Intermediate 46-2, substituting E6 for Intermediate E3 in Step 1.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.27H.sub.33F.sub.2N.sub.4O.sub.5: 531.24. found: 531.2.
[1130] Example 95 was prepared in a similar fashion to Example 62,
substituting Intermediate 95-1 for Intermediate 46-2 in Step 1 and
substituting Intermediate A10 for Intermediate A9 in Step 5.
Example 95 was isolated. Analytic HPLC RetTime: 8.86 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.42H.sub.52F.sub.4N.sub.7O.sub.8S: 890.35. found: 889.94.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.34 (s, 1H), 7.80 (m,
2H), 7.42 (d, J=8.6 Hz, 1H), 6.85 (s, 1H), 6.69 (s, 1H), 5.38 (m,
1H), 5.29 (m, 3H), 5.02 (d, J=8.8 Hz, 1H), 4.46 (d, J=7.4 Hz, 1H),
4.10-3.97 (m, 2H), 3.84 (d, J=7.9 Hz, 1H), 3.74 (d, J=8.6 Hz, 1H),
2.42-2.29 (m, 1H), 2.10 (s, 1H), 1.87-1.72 (m, 1H), 1.69-1.48 (m,
4H), 1.38 (d, J=14.8 Hz, 2H), 1.30-1.08 (m, 4H), 0.99 (s, 5H), 0.89
(m, 3H), 0.69 (s, 10H), 0.64 (m, 1H), 0.43 (s, 1H), 0.11 (m, 1H),
0.01 (m, 1H).
Example 96
Preparation of
(1aS,2aR,6S,9S,10S,11R,21E,24aR,24bS)-6-tert-butyl-15-chloro-N-[(1R,2R)-2-
-(difluoromethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-
-10-methyl-4,7,18-trioxo-1a,2,2a,4,5,6,7,10,11,20,23,24,24a,24b-tetradecah-
ydro-1H,9H,18H-8,11-methanocyclopropa[4',5]cyclopenta[1',2':18,19][1,10,3,-
6,12]dioxatriazacyclononadecino[11,12-b]quinazoline-9-carboxamide
##STR00368##
[1132] Example 96 was prepared in a similar fashion to Example 89,
substituting intermediate 96-1 for intermediate 17-4 in Step 1.
Intermediate 96-1 was prepared in a similar fashion to intermediate
17-4 of Example 17, substituting E9 for E3 and B1 for B4 in Step 1,
and substituting intermediate D16 for intermediate 011 in Step 3.
Example 96 was isolated. Analytic HPLC RetTime: 9.18 min.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.41H.sub.52C.sub.1F.sub.2N.sub.6O.sub.9S: 877.32. found:
877.61. .sup.1H NMR (400 MHz, Chloroform-d) .delta. 9.76 (s, 1H),
8.03 (d, J=8.6 Hz, 1H), 7.39 (m, 1H), 7.27 (m, 1H), 6.80 (s, 1H),
5.92 (m, 1H), 5.87-5.73 (m, 1H), 5.68 (m, 1H), 5.64-5.51 (m, 1H),
5.21 (m, 1H), 4.93 (m, 2H), 4.52-4.32 (m, 3H), 4.15-3.94 (m, 2H),
2.86-2.71 (m, 1H), 2.26 (m, 1H), 2.15 (m, 2H), 2.10-2.02 (m, 1H),
2.02-1.84 (m, 2H), 1.77 (m, 2H), 1.61 (s, 3H), 1.50 (m, 4H),
1.42-1.17 (m, 6H), 1.17-0.92 (m, 10H), 0.92-0.78 (m, 2H), 0.51-0.37
(m, 1H).
Example 97
Preparation of
(1aS,2aR,6S,9S,10S,11R,23aR,23bS)-6-tert-butyl-15-cyano-N-[(2R)-2-(difluo-
romethyl)-1-{[(1-methylcyclopropyl)sulfonyl]carbamoyl}cyclopropyl]-10-meth-
yl-4,7-dioxo-1a,2,2a,4,5,6,7,10,11,19,20,21,22,23,23a,23b-hexadecahydro-1H-
,9H-8,11-methanocyclopropa[4',5]cyclopenta[1',2':18,19][1,10,3,6]dioxadiaz-
acyclononadecino[11,12-b]quinoxaline-9-carboxamide
##STR00369## ##STR00370##
[1134] Intermediate 97-1 was prepared in a similar fashion to
intermediate 79-5, substituting E2 for E5 in Step 1. LCMS-ESI.sup.+
(m/z): [M+H].sup.+ calcd for C.sub.43H.sub.55N.sub.4O.sub.7:
739.41. found: 739.31.
[1135] Step 1. Preparation of 97-2. Macrocyclic olefin 97-1 (0.84
g, 1.14 mmol) was dissolved in 114 mL ethanol and 114 mL ethyl
acetate. After degassing with Argon, 0.84 g of 5% Pd/C Degussa-type
was added and the mixture was hydrogenated for 4 hours at 1 atm.
Filtration through celite, concentration, and silica gel
chromatography (40%-60% ethyl acetate in hexanes gradient) provided
intermediate 97-2. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.36H.sub.51N.sub.4O.sub.7: 651.38. found: 651.32.
[1136] Step 2. Preparation of 97-3. An ice cold solution of
macrocycle phenol 97-2 (0.47 g, 0.73 mmol) and triethylamine (0.81
ml, 5.81 mmol) in 3 mL DCM was treated with
trifluoromethanesulfonic anhydride solution, 1M in methylene
chloride (0.18 ml, 1.09 mmol) dropwise. After stirring for 2 hours,
the reaction was quenched with water and extracted with ethyl
acetate. The organic layer was washed with water and brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated. Silica gel
chromatography using a 5%-50% ethyl acetate in hexanes gradient
gave 97-3 as the first eluting peak (55 mg). LCMS-ESI.sup.+ (m/z):
[M+H].sup.+ calcd for C.sub.37H.sub.50F.sub.3N.sub.4O.sub.9S:
783.33. found: 782.96.
[1137] Step 3. Preparation of 97-4. Degassed a mixture of
macrocycle triflate 97-3 (408 mg, 0.52 mmol),
tetrakis(triphenylphosphine)palladium (30.11 mg, 0.03 mmol), Zinc
cyanide, 98% (61.21 mg, 0.52 mmol) in 2.6 mL DMF for 10 minutes.
The reaction was heated at 80.degree. C. for 1 hour. An additional
60 mg tetrakis(triphenylphosphine)palladium and 120 mg Zinc cyanide
were added and heating was continued for 30 minutes. The reaction
was quenched with saturated ammonium chloride solution and
extracted with ethyl acetate. The organic phase was separated,
dried over anhydrous sodium sulphate, filtered and concentrated.
The crude product was purified by silica gel chromatography using a
gradient of 5%-70% ethyl acetate in hexanes to give intermediate
97-4. LCMS-ESI.sup.+ (m/z): [M+H].sup.+ calcd for
C.sub.37H.sub.50N.sub.5O.sub.6: 660.38. found: 660.10.
[1138] Step 4. Preparation of 97-5. A solution of 97-4 (290 mg,
0.44 mmol) in 1 mL DCM was treated with 0.5 mL of TFA and stirred
overnight. Water was added and the mixture was extracted with ethyl
acetate. The organic phase was separated, dried over anhydrous
sodium sulphate, filtered and concentrated. The crude product was
purified by silica gel chromatography using a gradient of 10%-70%
ethyl acetate in hexanes to give intermediate 97-5 (216 mg) as a
white solid. LCMS-ESI (m/z): [M+H].sup.+ calcd for
C.sub.33H.sub.42N.sub.5O.sub.6: 604.31. found: 604.00.
[1139] Step 5. Preparation of Example 97. A mixture of 97-5 (50 mg,
0.08 mmol), HATU (37.79 mg, 0.1 mmol), in 0.3 mL DMF was stirred 5
min, then A10 (50 mg, 0.08 mmol) and DIPEA (0.06 ml, 0.33 mmol)
were added. After 45 min at rt, the reaction was incomplete (LCMS).
Added another 20 mg of A10 and stirred for 2 hours. 2 mL of 1N HCl
was added, and the mixture was extracted with DCM. The crude
product was purified by silica gel chromatography using a gradient
of 30%-65% ethyl acetate in hexanes. Combined product fractions
contained some residual DMF. Water was added, which generated a
precipate (14 mg). The filtrate was extracted with ethyl acetate,
and the extracts were combined with the precipitate. The resulting
solution was dried over anhydrous sodium sulphate, filtered,
concentrated and dried under reduced pressure to give Example 97.
Analytic HPLC RetTime: 9.06 min. LCMS-ESI (m/z): [M+H].sup.+ calcd
for C.sub.42H.sub.54F.sub.2N.sub.7O.sub.8S: 854.98. found: 853.88.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.77 (br s, 1H), 8.05 (m,
1H), 7.93 (m, 1H), 7.62 (m, 1H), 7.20 (m, 1H), 7.08 (m, 1H), 6-5.65
(m, 1H), 5.56 (m, 1H), 5.17 (m, 1H), 4.90 (m, 1H), 4.38 (m, 2H),
4.22 (m, 1H), 4.06 (m, 1H), 3.57 (m, 1H), 2.88 (m, 1H), 2.70 (m,
5H), 2.28-2.08 (m, 1H), 2.04-1.30 (m, 12H), 1.29-1.09 (m, 9H),
1.08-0.96 (m, 4H), 0.85-0.67 (m, 3H), 0.43 (m, 1H), 0.34 (m, 1H),
0.19-0.03 (m, 1H).
[1140] The following compounds can be made with the synthetic
methods of this disclosure, or by means generally well known in the
art:
##STR00371## ##STR00372## ##STR00373## ##STR00374## ##STR00375##
##STR00376## ##STR00377## ##STR00378## ##STR00379## ##STR00380##
##STR00381## ##STR00382## ##STR00383## ##STR00384## ##STR00385##
##STR00386## ##STR00387## ##STR00388## ##STR00389## ##STR00390##
##STR00391## ##STR00392## ##STR00393## ##STR00394## ##STR00395##
##STR00396## ##STR00397## ##STR00398## ##STR00399## ##STR00400##
##STR00401## ##STR00402## ##STR00403## ##STR00404## ##STR00405##
##STR00406## ##STR00407## ##STR00408## ##STR00409## ##STR00410##
##STR00411## ##STR00412## ##STR00413## ##STR00414## ##STR00415##
##STR00416## ##STR00417## ##STR00418## ##STR00419## ##STR00420##
##STR00421## ##STR00422## ##STR00423## ##STR00424## ##STR00425##
##STR00426## ##STR00427## ##STR00428## ##STR00429## ##STR00430##
##STR00431## ##STR00432## ##STR00433## ##STR00434## ##STR00435##
##STR00436## ##STR00437## ##STR00438## ##STR00439## ##STR00440##
##STR00441## ##STR00442## ##STR00443## ##STR00444## ##STR00445##
##STR00446## ##STR00447## ##STR00448## ##STR00449## ##STR00450##
##STR00451## ##STR00452## ##STR00453## ##STR00454## ##STR00455##
##STR00456## ##STR00457## ##STR00458## ##STR00459## ##STR00460##
##STR00461## ##STR00462## ##STR00463## ##STR00464## ##STR00465##
##STR00466## ##STR00467##
##STR00468## ##STR00469## ##STR00470## ##STR00471## ##STR00472##
##STR00473## ##STR00474## ##STR00475## ##STR00476## ##STR00477##
##STR00478## ##STR00479## ##STR00480## ##STR00481## ##STR00482##
##STR00483## ##STR00484## ##STR00485## ##STR00486## ##STR00487##
##STR00488## ##STR00489## ##STR00490## ##STR00491## ##STR00492##
##STR00493## ##STR00494## ##STR00495## ##STR00496## ##STR00497##
##STR00498## ##STR00499## ##STR00500## ##STR00501## ##STR00502##
##STR00503## ##STR00504## ##STR00505## ##STR00506## ##STR00507##
##STR00508## ##STR00509## ##STR00510## ##STR00511## ##STR00512##
##STR00513## ##STR00514## ##STR00515## ##STR00516## ##STR00517##
##STR00518## ##STR00519## ##STR00520## ##STR00521## ##STR00522##
##STR00523## ##STR00524## ##STR00525## ##STR00526## ##STR00527##
##STR00528## ##STR00529## ##STR00530## ##STR00531## ##STR00532##
##STR00533## ##STR00534## ##STR00535## ##STR00536## ##STR00537##
##STR00538## ##STR00539## ##STR00540## ##STR00541## ##STR00542##
##STR00543## ##STR00544## ##STR00545## ##STR00546## ##STR00547##
##STR00548## ##STR00549## ##STR00550## ##STR00551## ##STR00552##
##STR00553## ##STR00554## ##STR00555## ##STR00556## ##STR00557##
##STR00558## ##STR00559## ##STR00560## ##STR00561## ##STR00562##
##STR00563## ##STR00564## ##STR00565## ##STR00566## ##STR00567##
##STR00568## ##STR00569##
##STR00570## ##STR00571## ##STR00572## ##STR00573## ##STR00574##
##STR00575## ##STR00576## ##STR00577## ##STR00578## ##STR00579##
##STR00580## ##STR00581## ##STR00582## ##STR00583## ##STR00584##
##STR00585## ##STR00586## ##STR00587## ##STR00588## ##STR00589##
##STR00590## ##STR00591## ##STR00592## ##STR00593## ##STR00594##
##STR00595## ##STR00596## ##STR00597## ##STR00598## ##STR00599##
##STR00600## ##STR00601## ##STR00602## ##STR00603## ##STR00604##
##STR00605## ##STR00606## ##STR00607## ##STR00608## ##STR00609##
##STR00610## ##STR00611## ##STR00612## ##STR00613## ##STR00614##
##STR00615## ##STR00616## ##STR00617## ##STR00618## ##STR00619##
##STR00620## ##STR00621## ##STR00622## ##STR00623## ##STR00624##
##STR00625## ##STR00626## ##STR00627## ##STR00628## ##STR00629##
##STR00630## ##STR00631## ##STR00632## ##STR00633## ##STR00634##
##STR00635## ##STR00636## ##STR00637## ##STR00638## ##STR00639##
##STR00640## ##STR00641## ##STR00642## ##STR00643##
[1141] wherein V is a structure of Formula:
##STR00644##
and wherein E and G are defined as above.
Biological Activity
Expression and Purification of Genotype 1a, 2a, and 3 NS3
Proteases
Generation of NS3 Protease Expression Plasmids
[1142] The coding sequence of the genotype 1b (con-1 strain) HCV
NS3 protease domain was PCR amplified from a plasmid encoding the
I389luc-ubi-neo/NS3-3'/ET replicon (Reblikon, Mainz, Germany). The
5'-PCR primer was designed to encode an N-terminal K.sub.3
hexahistidine tag and to insert an in-frame recombinant Tobacco
Etch virus (rTEV) protease cleavage site into the NS3 coding
sequence. The resulting DNA fragment was cloned into the pET28
protein expression vector (Invitrogen, Carlsbad, Calif.) yielding
the p28-N6H-Tev-NS3(181)1b.
[1143] The coding sequences for the genotype 3 HCV protease domain
was amplified by RT-PCR using a Titan One Tube RT-PCR Kit (Roche,
Indianapolis, Ind.) and RNA extracted from HCV-positive human serum
(BBI Diagnostics, MA) using a QIAmp UltraSens Virus Kit (Qiagen,
Valencia, Calif.). 5' PCR primers were designed to encode
N-terminal hexahistidine tags and to insert in-frame rTEV protease
cleavage sites into the NS3 protease coding sequences. The
resulting DNA fragments were cloned into pET28 yielding the
expression vectors p28-N6H-Tev-NS3(181)1a and
p28-N6H-Tev-NS3(181)3, respectively.
NS3 Protease Protein Expression
[1144] BL21AI bacteria (Invitrogen, Carlsbad, Calif.) were
transformed with genotype 1b or 3 NS3 expression vectors and used
to inoculate a 20 L fermentation vessel (Sartorius BBI System Inc.,
Bethlehem, Pa.), containing 18 L of fresh 2YT medium supplemented
with 50 .mu.g/ml kanamycin. When cell densities reached an
OD.sub.600 of 1, the temperature of the cultures was reduced from
37.degree. C. to 280 C and induction was immediately initiated by
the addition of 30 .mu.M ZnSO.sub.4, 14 mM L-arabinose and 1 mM
Isopropyl .beta.-D-thiogalactoside (IPTG) final concentrations.
Cells were harvested by centrifugation four hours post-induction
and were stored as frozen pellets at -80.degree. C. prior to NS3
protein purification.
Purification of NS3 Proteases
Purification of Genotype 1b NS3 Protease
[1145] Cell pellets were thawed and resuspended at 10 ml/g cells in
lysis buffer containing 50 mM tris pH 7.6, 300 mM NaCl, 0.1%
3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS),
5% glycerol, and 2 mM .beta.-mercaptoethanol. Cell suspensions were
then sonicated, filtered through cheesecloth, and passed three
times through a microfluidizer at 18,000 pounds/in.sup.2. The
resulting lysates were centrifuged at 15500 rpm for 45 minutes and
supernatants were loaded onto a HisTrap HP column (GE Lifesciences)
pre-equilibrated with five volumes of Ni buffer A (50 mM tris pH
7.6, 300 mM NaCl, 0.1% CHAPS, 5% glycerol, 2 mM
.beta.-mercaptoethanol, 50 mM imidazole-HCl). Proteins were eluted
with a 0-100% gradient of Ni buffer A plus 500 mM imidazole-HCl and
fractions were collected and pooled. The HisTrap pool was diluted
1:10 with SP-A buffer (50 mM tris pH 7.0, 10% glycerol, 2 mM
dithiothreitol (DTT)) and loaded onto a HiTrap SP-HP column (GE
Lifesciences) equilibrated with SP-A buffer. NS3 protease was
eluted with a 0-100% SP-B buffer (SP-A buffer plus 1 M NaCl)
gradient. Concentrated pools of NS3-containing SP fractions were
aliquoted, snap frozen in liquid nitrogen and stored at -80.degree.
C.
Purification of Genotype 3 NS3 Protease
[1146] Bacterial pellets collected from the expression of genotype
3 HCV NS3 protease were homogenized in Lysis Buffer (25 mM tris, pH
7.5 buffer containing 150 mM NaCl and 1 mM phenylmethanesulfonyl
fluoride (PMSF)) and passed through a microfluidizer at 18,000
pounds/in.sup.2. Homogenized cell lysates were centrifuged at
30,000.times.g for 30 minutes at 4.degree. C. The resulting P1
pellets were washed with Wash Buffer I (25 mM tris, pH 7.5
containing 1% CHAPS) followed by centrifugation at 10,000.times.g
for 30 minutes at 4.degree. C. The resulting P2 pellets were washed
with Wash Buffer II (50 mM CAPS buffer, pH 10.8, containing 2M NaCl
and 2M urea) followed by centrifugation at 30,000.times.g for
minutes at 4.degree. C. The resulting P3 pellets were resuspended
in Solubilization Buffer (20 ml of 25 mM tris, pH 7.5 containing
150 mM NaCl and 8 M urea) and incubated at 4.degree. C. for one
hour. Solubilized proteins were passed through a 0.45 micron
filter. Protein concentrations were measured and the solutions were
adjusted to 40 mM DTT, incubated for 30 minutes at 4.degree. C. and
then quickly diluted into Refolding Buffer (25 mM tris, pH 8.5, 0.8
M Guanidine-HCl, 0.4 M L-Arginine, 10 mM ZnSO.sub.4) while
stirring. Protein solutions were incubated at 4.degree. C.
overnight to allow refolding. Refolded proteases were centrifuged
at 30,000.times.g for 10 minutes to remove residual precipitates.
Final protein concentrations were then measured and the NS3
proteases were aliquoted, snap frozen in liquid nitrogen and stored
at -80.degree. C.
Ki Determination for Genotypes 1b and 3a NS3 Protease.
[1147] Purified NS3 protease domain (amino acids 1-181) of the
genotype 1b and 3a virus were generated as above. The internally
quenched fluorogenic depsipeptide substrate
Ac-DED(Edans)-EEAbu.PSI.[COO]ASK(Dabcyl)-NH.sub.2 and a synthetic
peptide containing the hydrophobic core residues of the NS4A
protein cofactor (KKGSVVIVGRIILSGRKK; NS4A peptide) were obtained
from Anaspec, Inc. (San Jose, Calif.). Other chemicals and
biochemicals were of reagent grade or better and were purchased
from standard suppliers.
[1148] Reactions were run at room temperature in buffer consisting
of 50 mM HEPES, 40% glycerol, 0.05% Triton X-100, 10 mM DTT, and
10% DMSO. The final assay solutions contained 50 .mu.M NS3 genotype
1b protease or 200 .mu.M genotype 3a protease, 20 .mu.M NS4A
peptide, and 4 .mu.M substrate (genotype 1b) or 2 .mu.M substrate
(genotype 3a). Inhibitor concentrations varied from 100 nM to 5
.mu.M in 3-fold dilutions, and no-inhibitor controls were
included.
[1149] Compound dilutions were made in DMSO at 20.times. final
concentration. Reaction mixtures were prepared in 96-well assay
plates. A solution of enzyme and NS4A peptide in assay buffer (25
.mu.L volume with both reagents at 4.times. final concentration)
was mixed with 45 .mu.L assay buffer and 5 .mu.L of either
inhibitor or DMSO, and pre-incubated at room temperature for 1
hour. The reaction was started by addition of 25 .mu.L substrate
solution at 4.times. final concentration. Plates were mixed
vigorously for 5-10 seconds and reactions were allowed to proceed
for 90 minutes. fluorescence was measured every 30 s between 90 and
120 minutes reaction time using a Tecan InfiniTe M1000 or
PerkinElmer Envision multimode plate reader with an excitation
wavelength of 340 nm and an emission wavelength of 490 nm.
[1150] Rates were calculated from the progress curves at steady
state, in the time frame of 90-120 minutes after addition of
substrate. To determine the K, rates were plotted as a function of
inhibitor concentration, and the data were fit with equation 1
(Morrison, J. F., Biochimica et Biophysica Acta 1969, 185, 269-286)
to calculate K.sub.i.sup.app using GraphPad Prism 5. Active
fraction of enzyme was determined by active site titration with
known potent inhibitors. K was calculated from
K.sub.i.sup.app/(1+[[S]/K.sub.m]). Ki results for representative
compounds for genotype 1b and 3a (Ki 1B and Ki 3A, respectively)
are reported in Table 1.
v v 0 = [ E ] t - [ I ] t - K i app + ( [ E ] t - [ I ] t - K i app
) 2 + 4 [ E ] t K i app 2 [ E ] t ( 1 ) ##EQU00001##
Evaluation of Cell-Based Anti-HCV Activity:
[1151] Antiviral potency (EC.sub.50) was determined in both stable
subgenomic HCV replicon cell lines and transient-transfected HCV
replicon cells. The term half maximal effective concentration
(EC.sub.50) refers to the concentration of a drug which induces a
response halfway between the baseline and maximum after the
exposure time specified below.
[1152] Stable subgenomic HCV replicons for genotype 1a, 1b, 2a, 3a,
and 4a were established in Huh-7-derived cells as described by
Lohmann et al (Lohmann V, Korner F, Koch J, et al Replication of
subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science
1999; 285:119-3). Each stable cell line contains a bicistronic HCV
replicon that encodes a humanized Renilla luciferase (hRLuc)
reporter gene fused to a selectable neomycin-resistance gene,
followed by an EMCV IRES and the NS3-NS5B coding region of HCV.
Selection for cells constitutively expressing the HCV replicon was
achieved in the presence of the selection antibiotic, neomycin
(G418). Luciferase activity was measured as a marker for
intracellular HCV replication levels.
[1153] The genotype 1a stable replicon was derived from the H77 HCV
strain and contained adaptive mutations P1496L and S2204I. The
genotype 1b stable replicon was derived from the Con1 HCV strain
and contained adaptive mutations E1202G, T1280I, and K1846T. The
genotype 2a stable replicon was derived from the JFH-1 HCV strain
and did not require adaptive mutations. The genotype 3a stable
replicon was derived from the S52 HCV strain and contained adaptive
mutations P1121L, A1198T and S2210I (equivalent to S2204I in
genotype 1). The genotype 4a stable replicon was derived from the
ED43 HCV strain and contained adaptive mutations Q1691R and S2204I.
All replicon cell lines were propagated in Huh-7-derived cells and
maintained in Dulbecco's modified Eagle's Medium (DMEM)
supplemented with 10% fetal bovine serum (FBS) and 0.5 mg/ml
G418.
[1154] Transient-transfected HCV replicons were established for
genotype 1a, 1b, 3a and NS3/4a protease inhibitor resistant
variants D168A in genotype 1b or R155K in genotype 1a.
Transient-transfected replicons are also biscistronic subgenomic
replicons but do not contain the neomycin selectable marker present
in stable replicons. These replicons encode the poliovirus IRES
followed by the hRLuc reporter gene, the EMCV IRES and finally the
NS3-NS5B coding region of HCV. The genotype 1a (H77) and 1b (Con1)
wild-type replicons were derived from the same strain and contained
the same adaptive mutations as listed above. The genotype 3a
transient replicon was derived from the S52 HCV strain as above,
but contained slightly different adaptive mutations P1112L, K1615E
and S2210I. Specifically, the secondary adaptive mutation A1198T
(A166T) in the protease domain of the stable genotype 3a replicon
was replaced with K1615E (K583E) in the NS3 helicase, with no
effect on replication efficiency. Removal of A166T located in the
protease domain minimizes the impact of this variant on inhibitors
targeting the protease domain and represents a protease domain
closer to wild type for genotype 3a. Resistant replicons encoding
NS3/4 protease inhibitor mutations were introduced into the 1b or
1a wild-type NS3 gene by site directed mutagenesis. In vitro
transcribed RNAs from all transient replicons were transfected into
naive Huh-7-derived cell lines by electroporation. Luciferase
activity was measured as a marker for intracellular HCV replication
levels
[1155] To perform EC.sub.50 assays, cells from each HCV replicon
were dispensed into 384-well plates. Compounds were dissolved in
DMSO at a concentration of 10 mM and diluted in DMSO using an
automated pipetting instrument. Three-fold serially diluted
compounds were directly added to the cells using an automated
instrument. DMSO was used as a negative (solvent; no inhibition)
control, and a combination of three HCV inhibitors including a
protease inhibitor; an NS5A inhibitor and a nucleoside inhibitor
was used at concentrations >100.times. EC.sub.50 as a positive
control (100% inhibition). Seventy-two hours later, cells were
lysed and Renilla luciferase activity were quantified as
recommended by the manufacturer (Promega-Madison, Wis.). Non-linear
regression was performed to calculate EC.sub.50 values.
Results are shown in Tables 1 and 2:
TABLE-US-00001 TABLE 1 Biological Activity Values For Stable
Subgenonic HCV Replicon Cell Lines EC.sub.50 1A EC.sub.50 1B
EC.sub.50 2A EC.sub.50 3A EC.sub.50 4A Ki 1B Ki 3A RLUC* RLUC*
RLUC* RLUC* RLUC* Example (nM) (nM) (nM) (nM) (nM) (nM) (nM) 1 0.03
0.07 4.4 3.9 4.1 46 3.1 2 0.01 0.04 4.0 3.1 3.9 77 2.7 3 0.18 0.56
11.7 9.8 28 546 10 4 0.17 0.56 10.7 9.6 16 271 7.9 5 0.04 0.17 8.7
7.4 11 405 6.9 6 0.20 0.62 35 36 34 1361 34 7 0.05 0.06 4.9 3.8 4.2
67 3.2 8 0.07 0.42 16 8.6 20 465 13 9 0.15 0.59 17 7.9 23 1268 11
10 0.16 0.52 30 22 49 978 26 11 0.23 0.88 28 17 34 1162 19 12 0.27
1.2 34 18 25 2013 21 13 0.04 0.18 13 9.5 26 685 11 14 0.07 0.24 9.7
6.8 7.0 308 7.3 15 0.05 0.30 11 6.8 9.8 550 7.8 16 0.09 0.21 12 7.4
6.2 201 8.1 17 0.04 0.06 3.9 3.3 3.7 15 2.9 18 0.03 0.10 3.9 2.6
5.0 70 2.8 19 0.02 0.13 4.0 2.6 4.6 89 3.1 20 0.12 0.53 8.1 5.2 19
392 5.9 21 0.10 0.45 6.8 4.7 12 263 6.2 22 0.07 1.3 15 7.5 27 727
11 23 0.08 1.1 13 7.5 23 587 9.9 24 0.05 0.92 12 7.5 20 663 9.2 25
0.05 0.39 8.8 6.3 13 409 7.1 26 0.05 0.17 6.3 4.3 12 297 6.0 27
0.03 0.08 6.6 5.2 6.7 266 6.1 28 0.03 0.08 6.7 4.6 6.2 266 4.1 29
0.06 0.12 10 8.9 11 137 7.2 30 0.14 0.63 55 35 47 2437 31 31 0.13
4.9 40 18 63 2071 20 32 0.18 0.87 59 30 35 2311 30 33 0.03 0.06 7.6
2.8 4.9 16 2.3 34 0.10 0.28 12 13 18 322 9.7 35 0.07 0.23 9.1 11
7.4 162 7.6 36 0.10 1.7 23 14 53 585 9.1 37 0.10 0.19 24 22 16 575
15 38 0.03 0.70 7.8 4.2 5.4 151 5.5 39 0.08 0.20 34 39 41 321 26 40
0.08 0.18 25 22 48 360 18 41 0.18 0.79 135 142 106 2606 135 42 0.10
0.75 20 16 17 343 14 43 0.06 0.16 4.9 3.8 5.2 92 3.6 44 0.03 0.08
3.1 2.1 2.9 53 2.2 45 0.04 0.48 21 8.3 24 549 13 46 0.03 0.05 3.1
2.7 3.8 17 2.5 47 0.07 0.19 3.8 3.6 12 58 4.2 48 0.07 0.09 2.0 1.8
9.7 73 2.2 49 0.04 0.07 3.7 4.1 5.2 20 3.5 50 63 100 4444 4444 379
19708 4444 51 0.31 0.84 40 39 103 1221 30 52 0.25 1.3 195 245 380
2307 161 53 0.03 0.09 51 7.7 13 18 2.8 54 0.11 0.50 33 18 76 260 13
55 0.09 0.20 13 3.8 12 140 3.5 56 0.38 1.0 41 37 57 1026 57 57 0.07
0.28 12 11 21 166 4.9 58 0.07 0.17 12 9.7 12 134 12 59 0.04 0.06
5.4 11 13 20 5.6 60 0.04 0.08 11 4.8 7.7 45 5.4 61 0.06 0.09 13 10
8.7 28 11 62 0.04 0.03 3.4 3.0 5.0 8.5 2.4 63 0.03 0.01 4.2 2.9 3.2
11 3.2 64 0.07 1.2 100 38 48 671 34 65 0.08 0.07 12 8.4 7.7 30 8.1
66 0.04 0.06 37 20 105 1786 25 67 0.04 0.32 11 12 24 383 12 68 0.05
0.63 13 7.7 18 364 9.4 69 0.07 0.17 5.2 4.9 11 64 5.8 70 0.05 0.05
8.4 8.5 15 41 6.4 71 0.03 0.14 6.5 5.3 23 160 5.3 72 0.05 4.1 365
300 740 1819 383 73 0.11 0.82 7.9 7.5 22 178 7.2 74 0.03 0.12 8.0
5.0 18 374 8.3 75 0.03 0.12 9.9 6.4 18 240 10 76 0.03 0.06 2.4 2.2
2.4 9.0 1.9 77 nt nt 23 12 29 741 16 78 0.21 0.82 267 394 195 1115
225 79 0.06 0.06 7.0 5.8 4.1 71 5.6 80 0.49 13 1127 344 748 44444
1182 81 0.04 0.05 4.0 3.6 3.6 12 4.3 82 0.03 0.04 8.5 9.3 4.7 25 10
83 0.09 0.48 62 58 19 1219 42 84 0.04 0.08 5.9 5.7 7.2 27 6.4 85
0.05 0.03 16 17 7.7 304 9.2 86 0.04 0.07 5.6 5.5 5.9 42 5.7 87 0.02
0.05 4.8 3.9 3.3 14 3.8 88 0.02 0.29 9.3 4.6 2.7 105 4.1 89 0.03
0.05 6.0 6.1 4.3 19 4.6 90 0.06 0.13 12 9.6 8.8 114 10.4 91 nt nt
2.4 1.9 4.0 149 2.4 92 nt nt 3.3 3.2 4.6 58 3.2 93 nt nt 12 14 15
116 11 94 nt nt 3.8 5.2 3.3 32 3.0 96 nt nt 12 11 5.4 77 8.9 97 nt
nt 6.3 4.6 5.8 67 5.3 nt--not tested *RULC: Renilla Luciferase
TABLE-US-00002 TABLE 2 Biological Activity Values For
Transient-Transfected HCV Replicon Cell Lines EC.sub.50 3A
EC.sub.501A EC.sub.50 1A EC.sub.501B EC.sub.501B WT* WT*
R155K.sup..dagger. WT* D168A.sup..dagger-dbl. (nM) (nM) (nM) (nM)
(nM) 1 17 3.2 4.9 2.2 17 2 nt 2.4 7.0 1.0 33 3 nt 6.6 32 3.5 128 4
68 7.1 20 6.5 66 5 nt 3.7 17 3.2 91 6 nt 26 58 11 216 7 14 3.8 6.7
2.3 20 8 nt 8.8 28 4.8 89 9 nt 12 208 2.5 360 10 nt 37 131 10 493
11 nt 20 159 9.4 605 12 nt 14 283 5.7 640 13 nt 8.6 59 3.1 209 14
nt 7.4 21 4.0 99 15 nt 6.5 20 3.0 182 16 nt 9.4 22 5.8 61 17 6.1
2.9 2.8 1.7 4.3 18 nt 2.3 5.0 1.2 24 19 nt 2.1 3.4 1.1 28 20 Nt 3.4
17 2.7 90 21 nt 4.1 15 3.8 70 22 nt 8.6 48 2.6 242 23 nt 9.5 36 3.6
173 24 nt 9.3 49 3.2 284 25 nt 4.4 17 3.6 116 26 nt 3.6 12 1.9 109
27 nt 6.0 20 4.3 70 28 nt 3.0 9 3.4 54 29 nt 4.8 11 3.1 48 30 nt 41
296 31 503 31 nt 27 154 6.6 805 32 nt 44 547 14 653 33 5.3 2.6 2.5
1.6 4.2 34 46 15 18 9 64 35 35 12 17 10 38 36 128 16 271 9.4 333 37
69 29 51 22 159 38 nt 4.5 8.4 2.8 25 39 89 23 63 16 105 40 156 17
74 8.6 129 41 539 164 505 154 715 42 nt 17 35 10 109 43 nt 3.8 8.7
2.4 41 44 7.0 2.4 4.0 1.4 15 45 nt 13 35 5.3 88 46 6.788 2.4 3.2
1.2 4.5 47 17 5.4 6.1 2.0 12 48 13 1.7 4.0 1.1 6.5 49 6.3 3.5 3.8
2.8 4.2 50 26825 4444 3830 4444 4444 51 265 28 92 24 318 52 538 150
516 161 887 53 15 5.3 5.6 2.0 11 54 147 19 27 10 123 55 71 8.0 32
2.3 53 56 226 63 168 59 252 57 54 12 17 7.5 48 58 38 12 18 14 23 59
15 10 6.8 6.8 6.4 60 9.8 5.8 8.6 2.3 15 61 13 12 10 9.3 6.7 62 4.0
3.5 2.7 1.5 2.0 63 6.9 4.1 4.0 2.1 3.4 64 256 37 50 17 104 65 17
9.4 8.1 6.4 11 66 735 35 240 14 396 67 107 14 42 10 86 68 139 14 37
4.4 78 69 42 8.2 15 3.9 28 70 17 7.7 5.4 6.1 7.3 71 49 9.1 30 3.8
66 72 642 600 227 165 687 73 45 8.8 25 6.2 75 74 138 8.8 44 2.1 56
75 56 14 45 3.5 51 76 3.4 2.0 2.1 1.0 2.8 77 472 21 34 5.0 80 78
194 189 225 156 248 79 9.2 6.1 7.1 3.0 11 80 nt 403 2862 53 443 81
4.3 3.7 2.5 2.4 2.7 82 16 7.8 6.2 4.1 5.7 83 300 62 133 27 202 84
11 5.4 4.0 2.5 5.2 85 101 12 22 7.0 57 86 16 4.0 3.7 3.4 10 87 7.7
2.9 2.8 1.3 4.0 88 35 5.0 14 5.5 24 89 5.5 6.0 3.7 3.2 5.1 90 43 nt
nt nt nt 91 25 2.3 3.5 1.3 9.2 92 8.0 3.0 3.0 1.7 5.3 93 26 13 13
14 39 94 10 3.2 3.1 1.9 9.2 96 12 5.2 3.8 4.1 3.6 97 5.8 3.6 3.7
2.5 8.8 nt: not tested *WT = wild type .sup..dagger.NS3/4a protease
inhibitor resistant variants R155K in genotype 1a
.sup..dagger-dbl.NS3/4a protease inhibitor resistant variants D168A
in genotype 1b
[1156] The data in Tables 1 and 2 represent an average over time of
each assays for each compound. For certain compounds, multiple
assays have been conducted over the life of the project. Thus, the
data reported in Tables 1 and 2 include the data reported in the
priority document, as well as data generated in the intervening
period.
Pharmaceutical Compositions
[1157] The following illustrate representative pharmaceutical
dosage forms, containing a compound of Formulas I, II, III, or IV
(such as any one of IVa-IVh) (`Compound X`), for therapeutic or
prophylactic use in humans.
TABLE-US-00003 (i) Tablet 1 mg/tablet Compound X = 100.0 Lactose
77.5 Povidone 15.0 Croscarmellose sodium 12.0 Microcrystalline
cellulose 92.5 Magnesium stearate 3.0 300.0
TABLE-US-00004 (ii) Tablet 2 mg/tablet Compound X = 20.0
Microcrystalline cellulose 410.0 Starch 50.0 Sodium starch
glycolate 15.0 Magnesium stearate 5.0 500.0
TABLE-US-00005 (iii) Capsule mg/capsule Compound X = 10.0 Colloidal
silicon dioxide 1.5 Lactose 465.5 Pregelatinized starch 120.0
Magnesium stearate 3.0 600.0
TABLE-US-00006 (iv) Injection (1 mq/ml) mq/ml Compound X = (free
acid form) 1.0 Dibasic sodium phosphate 12.0 Monobasic sodium
phosphate 0.7 Sodium chloride 4.5 1.0 N Sodium hydroxide solution
q.s. (pH adjustment to 7.0-7.5) Water for injection q.s. ad 1
mL
[1158] The above formulations may be obtained by conventional
procedures well known in the pharmaceutical art.
[1159] All references, including publications, patents, and patent
documents are incorporated by reference herein, as though
individually incorporated by reference. The invention has been
described with reference to various specific and preferred
embodiments and techniques. However, it should be understood that
many variations and modifications may be made while remaining
within the spirit and scope of the invention.
[1160] The use of the terms "a" and "an" and "the" and similar
references in the context of this disclosure (especially in the
context of the following claims) are to be construed to cover both
the singular and the plural, unless otherwise indicated herein or
clearly contradicted by context. All methods described herein can
be performed in any suitable order unless otherwise indicated
herein or otherwise clearly contradicted by context. The use of any
and all examples, or exemplary language (e.g., such as, preferred,
preferably) provided herein, is intended merely to further
illustrate the content of the disclosure and does not pose a
limitation on the scope of the claims. No language in the
specification should be construed as indicating any non-claimed
element as essential to the practice of the present disclosure.
[1161] Alternative embodiments of the claimed disclosure are
described herein, including the best mode known to the inventors
for practicing the claimed invention. Of these, variations of the
disclosed embodiments will become apparent to those of ordinary
skill in the art upon reading the foregoing disclosure. The
inventors expect skilled artisans to employ such variations as
appropriate (e.g., altering or combining features or embodiments),
and the inventors intend for the invention to be practiced
otherwise than as specifically described herein.
[1162] Accordingly, this invention includes all modifications and
equivalents of the subject matter recited in the claims appended
hereto as permitted by applicable law. Moreover, any combination of
the above described elements in all possible variations thereof is
encompassed by the invention unless otherwise indicated herein or
otherwise clearly contradicted by context.
[1163] The use of individual numerical values is stated as
approximations as though the values were preceded by the word
"about" or "approximately." Similarly, the numerical values in the
various ranges specified in this application, unless expressly
indicated otherwise, are stated as approximations as though the
minimum and maximum values within the stated ranges were both
preceded by the word "about" or "approximately." In this manner,
variations above and below the stated ranges can be used to achieve
substantially the same results as values within the ranges. As used
herein, the terms "about" and "approximately" when referring to a
numerical value shall have their plain and ordinary meanings to a
person of ordinary skill in the art to which the disclosed subject
matter is most closely related or the art relevant to the range or
element at issue. The amount of broadening from the strict
numerical boundary depends upon many factors. For example, some of
the factors which may be considered include the criticality of the
element and/or the effect a given amount of variation will have on
the performance of the claimed subject matter, as well as other
considerations known to those of skill in the art. As used herein,
the use of differing amounts of significant digits for different
numerical values is not meant to limit how the use of the words
"about" or "approximately" will serve to broaden a particular
numerical value or range. Thus, as a general matter, "about" or
"approximately" broaden the numerical value. Also, the disclosure
of ranges is intended as a continuous range including every value
between the minimum and maximum values plus the broadening of the
range afforded by the use of the term "about" or "approximately."
Thus, recitation of ranges of values herein are merely intended to
serve as a shorthand method of referring individually to each
separate value falling within the range, unless otherwise indicated
herein, and each separate value is incorporated into the
specification as if it were individually recited herein.
[1164] It is to be understood that any ranges, ratios and ranges of
ratios that can be formed by, or derived from, any of the data
disclosed herein represent further embodiments of the present
disclosure and are included as part of the disclosure as though
they were explicitly set forth. This includes ranges that can be
formed that do or do not include a finite upper and/or lower
boundary. Accordingly, a person of ordinary skill in the art most
closely related to a particular range, ratio or range of ratios
will appreciate that such values are unambiguously derivable from
the data presented herein.
* * * * *