U.S. patent application number 12/174820 was filed with the patent office on 2009-02-05 for substituted bicyclolactam compounds.
This patent application is currently assigned to Pfizer Inc. Invention is credited to Robert L. Dow, Michael J. Munchhof.
Application Number | 20090036425 12/174820 |
Document ID | / |
Family ID | 39926534 |
Filed Date | 2009-02-05 |
United States Patent
Application |
20090036425 |
Kind Code |
A1 |
Dow; Robert L. ; et
al. |
February 5, 2009 |
SUBSTITUTED BICYCLOLACTAM COMPOUNDS
Abstract
The invention provides compounds of formula (1), ##STR00001##
and the pharmaceutically acceptable salt thereof, wherein R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.5a, R.sup.5b, R.sup.5c, R.sup.5d,
Q, A, Z, and R.sup.7 are as described herein; compositions thereof;
and uses thereof.
Inventors: |
Dow; Robert L.; (Groton,
CT) ; Munchhof; Michael J.; (Salem, CT) |
Correspondence
Address: |
PFIZER INC.
PATENT DEPARTMENT, MS8260-1611, EASTERN POINT ROAD
GROTON
CT
06340
US
|
Assignee: |
Pfizer Inc
|
Family ID: |
39926534 |
Appl. No.: |
12/174820 |
Filed: |
July 17, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60953507 |
Aug 2, 2007 |
|
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12174820 |
|
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Current U.S.
Class: |
514/211.05 ;
540/490 |
Current CPC
Class: |
A61P 7/00 20180101; A61P
1/16 20180101; C07D 498/04 20130101; A61P 3/04 20180101; A61P 3/06
20180101; A61P 9/04 20180101; A61P 9/10 20180101; A61P 9/12
20180101; A61P 13/12 20180101; A61P 27/12 20180101; A61P 9/00
20180101; A61P 19/06 20180101; A61P 3/10 20180101; A61P 43/00
20180101; A61P 3/00 20180101; A61P 35/00 20180101; A61P 25/00
20180101 |
Class at
Publication: |
514/211.05 ;
540/490 |
International
Class: |
A61K 31/553 20060101
A61K031/553; C07D 267/08 20060101 C07D267/08; A61P 3/04 20060101
A61P003/04; A61P 3/10 20060101 A61P003/10 |
Claims
1. A compound having the formula ##STR00073## or a tautomer
thereof, or a pharmaceutically acceptable salt of said compound or
tautomer, wherein: (a) R.sup.1 is H --(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)perfluoroalkyl,
--(C.sub.1-C.sub.4)perfluoroalkoxy, or --(C.sub.1-C.sub.4)alkoxy;
(b) R.sup.2 and R.sup.3, taken separately, are independently H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; or
R.sup.2 and R.sup.3, taken together with the carbon to which they
are attached, is --(C.sub.3-C.sub.6)cycloalkyl; (c) R.sup.4 is H or
--(C.sub.1-C.sub.4)alkyl; (d) R.sup.5a, R.sup.5b, R.sup.5c and
R.sup.5d are each independently H, F, Cl, Br,
--(C.sub.1-C.sub.4)alkyl, --OH or --O--(C.sub.1-C.sub.4)alkyl; (e)
Q is --O-- or a bond; (f) A is a --(C.sub.3-C.sub.6)cycloalkylene
group, a --(C.sub.3-C.sub.6)cycloalkenylene group or phenylene; (g)
Z is --C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and
R.sup.6b are each independently --H or --(C.sub.1-C.sub.4)alkyl, or
R.sup.6a and R.sup.6b, taken together with the carbon to which they
are attached, is a --(C.sub.3-C.sub.6)cycloalkyl; (h) R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic; (i)
R.sup.8 is --OR.sup.9 or NHR.sup.10; (j) R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
(k) R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3.
2. The compound of claim 1, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein (a) R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are each
independently H or --CH.sub.3; (b) R.sup.5b and R.sup.5c are each
H; (c) R.sup.5d is H, F or Cl; (d) R.sup.5a is H, F, Cl or methyl;
(e) Z is --CH.sub.2-- or a bond, and (f) R.sup.7 is C(O)R.sup.8 or
cyano.
3. The compound of claim 2, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein: Q is a bond; and A is a --(C.sub.3-C.sub.10)cycloalkylene
group or a --(C.sub.3-C.sub.10)cycloalkenylene group.
4. The compound of claim 3, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein R.sup.5d is H.
5. The compound of claim 4, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein A is 1,4-cyclohexylene, cyclohex-3-en-1,4-di-yl,
tricyclo[3.2.1.0.about.2,4.about.]octylene-1,3-di-yl or
octahydropentalene-1,4-di-yl.
6. A compound of claim 5, having the formula ##STR00074## or a
tautomer thereof, or a pharmaceutically acceptable salt of said
compound or tautomer, wherein: R.sup.1 is H or --CH.sub.3; R.sup.2
is H or --CH.sub.3; R.sup.3 is H or --CH.sub.3; R.sup.4 is H or
--CH.sub.3; R.sup.5a is H, F, Cl or methyl; Z is
--C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and R.sup.6b
are each independently --H or --(C.sub.1-C.sub.4)alkyl, or R.sup.6a
and R.sup.6b, taken together with the carbon to which they are
attached, is a --(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8 is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3.
7. The compound of claim 6, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein Z is --CH.sub.2--.
8. The compound of claim 7, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein R.sup.7 is --C(O)NHR.sup.10.
9. The compound of claim 7, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein R.sup.7 is --CN.
10. The compound of claim 9 which is
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}acetonitrile or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer.
11. The compound of claim 7, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein R.sup.7 is --C(O)OH.
12. The compound of claim 11 which is
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-yl)--
2-fluorophenyl)cyclohexyl)acetic acid or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer.
13. The compound of claim 11 which is
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}acetic acid or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer.
14. The compound of claim 7, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer
wherein R.sup.2 is (R)-methyl.
15. The compound of claim 14, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein R.sup.7 is --CN.
16. The compound of claim 14 which is
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl}cyclohexyl)acetonitrile or a tautomer
thereof, or pharmaceutically acceptable salt of said compound or
tautomer.
17. The compound of claim 14, or a tautomer thereof, or
pharmaceutically acceptable salt of said compound or tautomer,
wherein R.sup.7 is --C(O)OH.
18. The compound of claim 17 which is
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl}cyclohexyl)acetic acid or a tautomer
thereof, or pharmaceutically acceptable salt of said compound or
tautomer.
19. The compound of claim 17 which is
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazep-
in-6(5H)-yl)-2-fluorophenyl)-cyclohexyl)acetic acid or a tautomer
thereof, or pharmaceutically acceptable salt of said compound or
tautomer.
20. A pharmaceutical composition comprising: (a) a compound having
the formula ##STR00075## or a tautomer thereof, or a
pharmaceutically acceptable salt of said compound or tautomer,
wherein: R.sup.1 is H --(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)perfluoroalkyl,
--(C.sub.1-C.sub.4)perfluoroalkoxy, or --(C.sub.1-C.sub.4)alkoxy;
R.sup.2 and R.sup.3, taken separately, are independently H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; or
R.sup.2 and R.sup.3, taken together with the carbon to which they
are attached, is --(C.sub.3-C.sub.6)cycloalkyl; R.sup.4 is H or
--(C.sub.1-C.sub.4)alkyl; R.sup.5a, R.sup.5b, R.sup.5c and R.sup.5d
are each independently H, F, Cl, Br, --(C.sub.1-C.sub.4)alkyl, --OH
or --O--(C.sub.1-C.sub.4)alkyl; Q is --O-- or a bond; A is a
--(C.sub.3-C.sub.6)cycloalkylene group, a
--(C.sub.3-C.sub.6)cycloalkenylene group or phenylene; Z is
--C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and R.sup.6b
are each independently --H or --(C.sub.1-C.sub.4)alkyl, or R.sup.6a
and R.sup.6b, taken together with the carbon to which they are
attached, is a --(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8 is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3; and (b) a pharmaceutically acceptable carrier,
vehicle, diluent or excipient.
21. The pharmaceutical composition of claim 20 comprising: (a) a
compound having the formula ##STR00076## or a tautomer thereof, or
a pharmaceutically acceptable salt of said compound or tautomer,
wherein: R.sup.1 is H or --CH.sub.3; R.sup.2 is H or --CH.sub.3;
R.sup.3 is H or --CH.sub.3; R.sup.4 is H or --CH.sub.3; R.sup.5a is
H, F, Cl or methyl; A is 1,4-cyclohexylene; Z is
--C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and R.sup.6b
are each independently --H or --(C.sub.1-C.sub.4)alkyl, or R.sup.6a
and R.sup.6b, taken together with the carbon to which they are
attached, is a --(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8 is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3; and (b) a pharmaceutically acceptable carrier,
vehicle, diluent or excipient.
22. A method for treating Type 2 diabetes in a mammal in need of
such treatment comprising administering to said mammal a
therapeutically effective amount of a compound having the formula
##STR00077## or a tautomer thereof, or a pharmaceutically
acceptable salt of said compound or tautomer, wherein: R.sup.1 is H
--(C.sub.1-C.sub.4)alkyl, --(C.sub.1-C.sub.4)perfluoroalkyl,
--(C.sub.1-C.sub.4)perfluoroalkoxy, or --(C.sub.1-C.sub.4)alkoxy;
R.sup.2 and R.sup.3, taken separately, are independently H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; or
R.sup.2 and R.sup.3, taken together with the carbon to which they
are attached, is --(C.sub.3-C.sub.6)cycloalkyl; R.sup.4 is H or
--(C.sub.1-C.sub.4)alkyl; R.sup.5a, R.sup.5b, R.sup.5c and R.sup.5d
are each independently H, F, Cl, Br, --(C.sub.1-C.sub.4)alkyl, --OH
or --O--(C.sub.1-C.sub.4)alkyl; Q is --O-- or a bond; A is a
--(C.sub.3-C.sub.6)cycloalkylene group, a
--(C.sub.3-C.sub.6)cycloalkenylene group or phenylene; Z is
--C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and R.sup.6b
are each independently --H or --(C.sub.1-C.sub.4)alkyl, or R.sup.6a
and R.sup.6b, taken together with the carbon to which they are
attached, is a --(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3.
23. A method of claim 22 for treating Type 2 diabetes in a mammal
in need of such treatment comprising administering to said mammal a
therapeutically effective amount of a compound having the formula
##STR00078## or a tautomer thereof, or a pharmaceutically
acceptable salt of said compound or tautomer, wherein: R.sup.1 is H
or --CH.sub.3; R.sup.2 is H or --CH.sub.3; R.sup.3 is H or
--CH.sub.3; R.sup.4 is H or --CH.sub.3; R.sup.5a is H, F, Cl or
methyl; A is 1,4-cyclohexylene; Z is --C(R.sup.6a)(R.sup.6b)-- or a
bond wherein R.sup.6a and R.sup.6b are each independently --H or
--(C.sub.1-C.sub.4)alkyl, or R.sup.6a and R.sup.6b, taken together
with the carbon to which they are attached, is a
--(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is C(O)R.sup.8, cyano,
hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8 is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3.
24. The method of claim 23 wherein said mammal is a human.
25. A method for treating obesity in a mammal in need of such
treatment comprising administering to said mammal a therapeutically
effective amount of a compound having the formula ##STR00079## or a
tautomer thereof, or a pharmaceutically acceptable salt of said
compound or tautomer, wherein: R.sup.1 is H
--(C.sub.1-C.sub.4)alkyl, --(C.sub.1-C.sub.4)perfluoroalkyl,
--(C.sub.1-C.sub.4)perfluoroalkoxy, or --(C.sub.1-C.sub.4)alkoxy;
R.sup.2 and R.sup.3, taken separately, are independently H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; or
R.sup.2 and R.sup.3, taken together with the carbon to which they
are attached, is --(C.sub.3-C.sub.6)cycloalkyl; R.sup.4 is H or
--(C.sub.1-C.sub.4)alkyl; R.sup.5a, R.sup.5b, R.sup.5c and R.sup.5d
are each independently H, F, Cl, Br, --(C.sub.1-C.sub.4)alkyl, --OH
or --O--(C.sub.1-C.sub.4)alkyl; Q is --O-- or a bond; A is a
--(C.sub.3-C.sub.6)cycloalkylene group, a
--(C.sub.3-C.sub.6)cycloalkenylene group or phenylene; Z is
--C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and R.sup.6b
are each independently --H or --(C.sub.1-C.sub.4)alkyl, or R.sup.6a
and R.sup.6b, taken together with the carbon to which they are
attached, is a --(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3.
26. A method of claim 25 for treating obesity in a mammal in need
of such treatment comprising administering to said mammal a
therapeutically effective amount of a compound having the formula
##STR00080## or a tautomer thereof, or a pharmaceutically
acceptable salt of said compound or tautomer, wherein: R.sup.1 is H
or --CH.sub.3; R.sup.2 is H or --CH.sub.3; R.sup.3 is H or
--CH.sub.3; R.sup.4 is H or --CH.sub.3; R.sup.5a is H, F, Cl or
methyl; A is 1,4-cyclohexylene; Z is --C(R.sup.6a)(R.sup.6b)-- or a
bond wherein R.sup.6a and R.sup.6b are each independently --H or
--(C.sub.1-C.sub.4)alkyl, or R.sup.6a and R.sup.6b, taken together
with the carbon to which they are attached, is a
--(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is C(O)R.sup.8, cyano,
hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8 is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3.
27. The method of claim 26 wherein said mammal is a human.
Description
FIELD OF THE INVENTION
[0001] The invention relates to substituted bicyclolactam
derivatives, pharmaceutical formulations thereof, and uses
thereof.
BACKGROUND OF THE INVENTION
[0002] Obesity, which is an excess of body fat relative to lean
body mass, is a chronic disease that is highly prevalent in modern
society. It is associated with decreased life span and numerous
medical problems, including adverse psychological development,
coronary artery disease, hypertension, stroke, Type 2 diabetes,
hyperlipidemia, and some cancers. A hallmark characteristic of
obesity is an increase in white adipose tissue (WAT) mass that is
largely due to accumulation of triacylglycerol. This increase in
WAT mass is a key contributor to obesity-associated
complications.
[0003] Diacylglycerol O-acyltransferase 1 (DGAT-1) is a
membrane-bound enzyme that catalyzes the terminal step of
triacylglycerol biosynthesis. DGAT-1 is expressed in the intestine
and adipose tissue It has been found that DGAT-1 null mice do not
become obese when challenged with a high fat diet in contrast to
wild-type littermates (Smith, et al., Nature Genetics 25:87 90,
2000). DGAT-1 null mice display reduced postprandial plasma glucose
levels and exhibit increased energy expenditure, but have normal
levels of serum triglycerides.
[0004] The phenotype of the DGAT-1 null mouse, along with the
results of our studies with DGAT-1 inhibitors in diet-induced obese
(DIO) mice, indicate that such mice are resistant to diet induced
obesity and have increased insulin and leptin sensitivity. These
effects suggest that inhibition of DGAT in vivo may be a novel
therapeutic target not only for obesity but also for diabetes.
(Subauste A, Burant C F, Curr Drug Targets Immune Endocr Metabol
Disord. December 2003; 3(4):263-70.)
[0005] Therefore, a need exits in the art, however, for DGAT-1
inhibitors that have efficacy for the treatment of metabolic
disorders such as, for example, obesity, Type 2 diabetes and
insulin resistance syndrome. Further, a need exists in the art for
DGAT-1 inhibitors having IC.sub.50 values of less than about 1000
nM and preferably below 100 nM.
SUMMARY OF THE INVENTION
[0006] The present invention relates to compounds having the
structure of formula (1)
##STR00002## [0007] or a tautomer thereof, or a pharmaceutically
acceptable salt of said compound or tautomer, wherein: [0008]
R.sup.1 is H --(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)perfluoroalkyl,
--(C.sub.1-C.sub.4)perfluoroalkoxy, or --(C.sub.1-C.sub.4)alkoxy;
[0009] R.sup.2 and R.sup.3, taken separately, are independently H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl;
[0010] or R.sup.2 and R.sup.3, taken together with the carbon to
which they are attached, is --(C.sub.3-C.sub.6)cycloalkyl; [0011]
R.sup.4 is H or --(C.sub.1-C.sub.4)alkyl; [0012] R.sup.5a,
R.sup.5b, R.sup.5c and R.sup.5d are each independently H, F, Cl,
Br, --(C.sub.1-C.sub.4)alkyl, --OH or --O--(C.sub.1-C.sub.4)alkyl;
[0013] Q is --O-- or a bond; [0014] A is a
--(C.sub.3-C.sub.6)cycloalkylene group, a
--(C.sub.3-C.sub.6)cycloalkenylene group or phenylene; [0015] Z is
--C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and R.sup.6b
are each independently --H or --(C.sub.1-C.sub.4)alkyl, or R.sup.6a
and R.sup.6b, taken together with the carbon to which they are
attached, is a --(C.sub.3-C.sub.6)cycloalkyl; [0016] R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)perfluoroalkoxy or a carboxylic acid mimic;
[0017] R.sup.8 is --OR.sup.9 or NHR.sup.10; [0018] R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
[0019] R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3.
[0020] The present invention also relates to a pharmaceutical
composition comprising a compound of the present invention, or a
pharmaceutically acceptable salt of the compound, and a
pharmaceutically acceptable carrier, vehicle, diluent or
excipient.
[0021] The present invention further relates to a method of
treating Type 2 diabetes, insulin resistance syndrome or obesity,
comprising administering to a mammal in need of such treatment a
therapeutically effective amount of a compound of the present
invention, or a pharmaceutically acceptable salt of the
compound.
[0022] The present invention additionally relates to a method of
inhibiting DGAT-1 in a mammal comprising administering to said
mammal an inhibitory amount of a compound of the present invention,
or a pharmaceutically acceptable salt of said compound.
[0023] The compounds, salts, and pharmaceutical compositions of the
present invention are useful for the treatment of obesity, Type 2
diabetes and insulin resistance syndrome.
[0024] The compounds, salts and pharmaceutical compositions of the
present invention are also useful for the treatment of impaired
glucose tolerance, hyperglycemia, diabetic complications such as
sugar cataracts, diabetic neuropathy, diabetic nephropathy,
diabetic retinopathy and diabetic cardiomyopathy, anorexia nervosa,
bulimia, cachexia, hyperuricemia, hyperinsulinemia,
hypercholesterolemia, hyperlipidemia, dyslipidemia, mixed
dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver
disease, atherosclerosis, arteriosclerosis, acute heart failure,
congestive heart failure, coronary artery disease, cardiomyopathy,
myocardial infarction, angina pectoris, hypertension, hypotension,
stroke, ischemia, ischemic reperfusion injury, aneurysm,
restenosis, vascular stenosis, solid tumors, skin cancer, melanoma,
lymphoma, breast cancer, lung cancer, colorectal cancer, stomach
cancer, esophageal cancer, pancreatic cancer, prostate cancer,
kidney cancer, liver cancer, bladder cancer, cervical cancer,
uterine cancer, testicular cancer and ovarian cancer.
DETAILED DESCRIPTION OF THE INVENTION
[0025] The terms used to describe the present invention have the
following meanings herein.
[0026] The compounds and intermediates of the present invention may
be named according to either the IUPAC (International Union for
Pure and Applied Chemistry) or CAS (Chemical Abstracts Service)
nomenclature systems.
[0027] The carbon atom content of the various
hydrocarbon-containing moieties herein may be indicated by a prefix
designating the minimum and maximum number of carbon atoms in the
moiety, for example, the prefixes (C.sub.a-C.sub.b)alkyl, and
C.sub.a-balkyl, indicate an alkyl moiety of the integer "a" to "b"
carbon atoms, inclusive. Thus, for example, (C.sub.1-C.sub.6)alkyl
and C.sub.1-6alkyl refer to an alkyl group of one to six carbon
atoms inclusive.
[0028] The symbol "--" represents a covalent bond.
[0029] The term "alkyl" denotes a straight or branched chain
monovalent radical of an aliphatic chain of carbon atoms. Examples
of alkyl groups include methyl, ethyl, propyl, isopropyl, isobutyl,
and the like.
[0030] The term "alkoxy" refers to straight or branched, monovalent
radical of a saturated aliphatic chain of carbon atoms bonded to an
oxygen atom that is attached to a core structure. Examples of
alkoxy groups include methoxy, ethoxy and iso-propoxy.
[0031] The term "carboxylic acid mimic` refers to a mimic or
bioisostere of carboxylic acid group as described in "The Practice
of Medicinal Chemistry", Wermuth C. G. Ed.; Academic Press; New
York, 1996, p 203. Examples of suitable carboxylic acid mimics
include, but are not limited to, --SO.sub.3H,
--CH.sub.2S(O).sub.2R.sup.7, --C(O)NHS(O).sub.2R.sup.7, --C(O)NHOH,
--C(O)NHCN, --C(O)NHR.sup.7, --CH(CF.sub.3)OH,
--C(CF.sub.3).sub.2OH, --P(O)(OH).sub.2,
1,2,5-thiadiazol-3-ol-4-yl, 1H-tetrazole-5-yl,
1H-1,2,4-triazole-5-yl, 1H-pyrazol-5-ol-3-yl, isoxazol-5-ol-3-yl,
isoxazol-3-ol-5-yl, thiazolidine-2,4-dione-5-yl,
imidazolidine-2,4-dione-5-yl, 1H-pyrrole-2,5-dione-3-yl,
1H-imidazol-2-ol-5-yl, dihydropyrimidine-2,4(1H,3H)-dione-6-yl,
imidazolidine-2,4-dione-1-yl, 1H-imidazol-5-ol-2-yl,
1H-pyrazol-3-ol-1-yl, 1H-pyrazol-3-ol-4-yl,
1,3,4-thiadiazol-2-ol-5-yl, 1,3,4-oxadiazol-2-ol-5-yl,
1,2,4-oxadiazol-3-ol-5-yl, 1,2,4-thiadiazol-3-ol-5-yl,
oxazol-2-ol-4-yl, thiazol-2-ol-4-yl, thiazol-4-ol-2-yl,
1,2,4-oxadiazol-5-ol-3-yl, 1,2,4-thiadiazol-5-ol-3-yl,
1,1-di-oxo-1,2,5-thiadiazolidin-3-one-2-yl,
1,1-di-oxo-1,2,5-thiadiazolidin-3-one-5-yl, isothiazol-3-ol-5-yl,
2H-1,2,3-triazol-4-ol-2-yl, 1H-1,2,3-triazol-4-ol-1-yl,
1H-imidazole-2,4-diol-5-yl,
1-oxo-2,3-dihydro-1,2,4-thiadiazol-5-ol-3-yl,
thiazole-2,4-diol-5-yl, oxazole-2,4-diol-5-yl,
3,4-dihydroxyfuran-2(5H)-one-5-yl, and
5-hydroxy-1,2,4-thiadiazol-3(2H)-one-2-yl. R.sup.7, as used herein,
is --H, --(C.sub.1-C.sub.4)alkyl or
--(C.sub.3-C.sub.6)cycloalkyl.
[0032] The term "cycloalkyl" denotes a non-aromatic, monocyclic
carbocyclic radical. Examples of cycloalkyl groups include
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. Cycloalkyl groups
may be optionally fused to aromatic hydrocarbons such as benzene to
form fused cycloalkyl groups, such as indanyl and the like.
[0033] The term "cycloalkylene" denotes a saturated, or partially
saturated, monocyclic, bicyclic or tricyclic carbocyclic di-valent
radical. Examples of cycloalkylene groups include cyclopropylene,
cyclobutylene, cyclopentylene, cyclohexylene, cyclohexen-di-yl
octahydropentalene-di-yl and tricycloctylene-di-yl. Cycloalkylene
groups may be optionally fused to aromatic hydrocarbons such as
benzene to form fused cycloalkylene groups, such as indanylene and
the like.
[0034] The term "perfluoroalkyl" is defined herein as a monovalent
alkyl radical wherein each hydrogen is substituted with a fluoro.
Examples of perfluoroalkyl groups include, but are not limited to,
trifluoromethyl, perfluoroethyl, and the like.
[0035] The term "perfluoroalkoxy" is defined herein as an alkoxy
group wherein each hydrogen is substituted with a fluoro. Examples
of perfluoroalkoxy groups include, but are not limited to,
trifluoromethoxyl, perfluoroethoxy, and the like.
[0036] The term "radical" denotes a group of atoms that behaves as
a single reactant in a chemical reaction, e. g., an organic radical
is a group of atoms that imparts characteristic properties to a
compound containing it, or which remains unchanged during a series
of reactions, or transformations.
[0037] The term "substituted" means that a hydrogen atom on a
molecule has been replaced with a different atom or molecule. The
atom or molecule replacing the hydrogen atom is denoted as a
"substituent."
[0038] The term "tautomer" refers to organic compounds that are
interconvertible by a chemical reaction called tautomerization.
Usually, the reaction involves the migration of a hydrogen atom or
proton, accompanied by a switch of a single bond and adjacent
double bond.
[0039] The terms "treating", "treated", or "treatment" as employed
herein includes preventing (e.g., prophylaxis), palliating, slowing
progression and curing a disease, such as obesity, insulin
resistance syndrome, Type 2 diabetes, or a disease-related
condition such as a diabetic complication.
[0040] The phrase "therapeutically effective amount" means an
amount of a compound of the present invention that (i) treats or
prevents the particular disease, condition, or disorder, (ii)
attenuates, ameliorates, or eliminates one or more symptoms of the
particular disease, condition, or disorder, or (iii) prevents or
delays the onset of one or more symptoms of the particular disease,
condition, or disorder described herein. The phrase
"pharmaceutically acceptable" indicates that the designated
carrier, vehicle, diluent, excipient(s), and/or salt is generally
chemically and/or physically compatible with the other ingredients
comprising the formulation, and physiologically compatible with the
recipient thereof.
[0041] The term "mammal" relates to an individual animal that is a
member of the taxonomic class Mammalia. Examples of mammals
include, but are not limited to, humans, dogs, cats, horses and
cattle. In the present invention, the preferred mammals are humans,
dogs and cats. More preferably, the mammal is a human.
[0042] The term "related salts" as used herein means
pharmaceutically acceptable salts of compounds of the present
invention.
[0043] In the present invention, it is preferred, for the compounds
of formula (1), or for tautomers thereof, or for salts of said
compounds or tautomers, that (a) R.sup.1, R.sup.2, R.sup.3 and
R.sup.4 are each independently H or --CH.sub.3; (b) R.sup.5b and
R.sup.5c are each H; (c) R.sup.5d is H, F or Cl; (d) R.sup.5a is H,
F, Cl or methyl; (e) Z is --CH.sub.2-- or a bond, and (f) R.sup.7
is C(O)R.sup.8 or cyano.
[0044] More preferably, Q is a bond; A is a
--(C.sub.3-C.sub.10)cycloalkylene group or a
--(C.sub.3-C.sub.10)cycloalkenylene group. R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are each independently H or --CH.sub.3;
R.sup.5b and R.sup.5c are each H; R.sup.5d is H, F or Cl; R.sup.5a
is H, F, Cl or methyl; Z is --CH.sub.2-- or a bond, and R.sup.7 is
C(O)R.sup.8 or cyano.
[0045] Even more preferably, R.sup.5d is H; Q is a bond; A is a
--(C.sub.3-C.sub.10)cycloalkylene group or a
--(C.sub.3-C.sub.10)cycloalkenylene group; R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are each independently H or --CH.sub.3;
R.sup.5b and R.sup.5c are each H; R.sup.5a is H, F, Cl or methyl; Z
is --CH.sub.2-- or a bond, and R.sup.7 is C(O)R.sup.8 or cyano.
[0046] Yet even more preferably A is 1,4-cyclohexylene,
cyclohex-3-en-1,4-di-yl,
tricyclo[3.2.1.0.about.2,4.about.]octylene-1,3-di-yl or
octahydropentalene-1,4-di-yl; R.sup.5d is H; Q is a bond; R.sup.1,
R.sup.2, R.sup.3 and R.sup.4 are each independently H or
--CH.sub.3; R.sup.5b and R.sup.5c are each H; R.sup.5a is H, F, Cl
or methyl; Z is --CH.sub.2-- or a bond, and R.sup.7 is C(O)R.sup.8
or cyano.
[0047] Most preferably, the compounds or formula (1), tautomers
thereof, and salts of the compounds or tautomers, include compounds
having the structure of formula (2)
##STR00003##
or a tautomer thereof, or a pharmaceutically acceptable salt of
said compound or tautomer, wherein: R.sup.1 is H or --CH.sub.3;
R.sup.2 is H or --CH.sub.3; R is H or --CH.sub.3; R.sup.4 is H or
--CH.sub.3; R.sup.5a is H, F, Cl or methyl; Z is
--C(R.sup.6a)(R.sup.6b)-- or a bond wherein R.sup.6a and R.sup.6b
are each independently --H or --(C.sub.1-C.sub.4)alkyl, or R.sup.6a
and R.sup.6b, taken together with the carbon to which they are
attached, is a --(C.sub.3-C.sub.6)cycloalkyl; R.sup.7 is
C(O)R.sup.8, cyano, hydroxyl, --(C.sub.1-C.sub.4)alkoxy,
--(C.sub.1-C.sub.4)-perfluoroalkoxy or a carboxylic acid mimic;
R.sup.8 is --OR.sup.9 or NHR.sup.10; R.sup.9 is --H,
--(C.sub.1-C.sub.4)alkyl, or --(C.sub.1-C.sub.4)perfluoroalkyl; and
R.sup.10 is --H, --(C.sub.1-C.sub.4)alkyl, tetrazolyl or
S(O).sub.2CF.sub.3. Preferably, Z is --CH.sub.2--.
[0048] In the present invention, it is preferred, for the compounds
of formula (2), or for tautomers thereof, or for salts of said
compounds or tautomers, that Z is --CH.sub.2--.
[0049] In one preferred embodiment of the compounds of formula (2),
or the tautomers thereof, or the salts of said compounds or
tautomers, Z is --CH.sub.2-- and R.sup.7 is --C(O)NHR.sup.10.
[0050] In a second preferred embodiment of the compounds of formula
(2), or the tautomers thereof, or the salts of said compounds or
tautomers, Z is --CH.sub.2-- and R.sup.7 is --CN. A prefered
compound of this embodiment is
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}acetonitrile, or a tautomer thereof, or a salt
of said compound or tautomer.
[0051] In a third preferred embodiment of the compounds of formula
(2), or the tautomers thereof, or the salts of said compounds or
tautomers, Z is --CH.sub.2-- and R.sup.7 is --C(O)OH. One prefered
compound of this embodiment is
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-yl)--
2-fluorophenyl)cyclohexyl)acetic acid, or a tautomer thereof, or a
salt of said compound or tautomer. Another prefered compound of
this embodiment is
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-oxazepin-6(-
5H)-yl)phenyl]cyclohexyl}acetic acid, or a tautomer thereof, or a
salt of said compound or tautomer.
[0052] In a fourth preferred embodiment of the compounds of formula
(2), or the tautomers thereof, or the salts of said compounds or
tautomers, Z is --CH.sub.2-- and R.sup.2 is (R)-methyl. For this
embodiment, it is more preferred that R.sup.7 is --C(O)NHR.sup.10,
--CN, or --C(O)OH. A prefered compound of this embodiment is
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl}cyclohexyl)acetonitrile, or a tautomer
thereof, or a salt of said compound or tautomer. Another prefered
compound of this embodiment is
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido-[5,4-f][1,4]-
oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetic acid, or a tautomer
thereof, or a salt of said compound or tautomer. Yet another
prefered compound of this embodiment is
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydro-pyrimido[5,4-f][1,4]oxaze-
pin-6(5H)-yl)-2-fluorophenyl)cyclohexyl)acetic acid, or a tautomer
thereof, or a salt of said compound or tautomer.
[0053] Wherein a compound of formula (I) contains an alkenyl or
alkenylene group, geometric cis/trans (or Z/E) isomers are
possible. Cis/trans isomers may be separated by conventional
techniques well known to those skilled in the art, for example,
chromatography and fractional crystallization.
[0054] When the compounds of the present invention contain one or
more stereogenic centers, said compounds may be resolved into the
pure enantiomers by methods known to those skilled in the art, for
example by formation of stereoisomeric salts which may be
separated, for example, by crystallization; formation of
stereoisomeric derivatives or complexes which may be separated, for
example, by crystallization, gas-liquid or liquid chromatography;
selective reaction of one enantiomer with an enantiomer-specific
reagent, for example enzymatic esterification; or gas-liquid or
liquid chromatography in a chiral environment, for example on a
chiral support for example silica with a bound chiral ligand or in
the presence of a chiral solvent. It will be appreciated that where
the desired stereoisomer is converted into another chemical entity
by one of the separation procedures described above, a further step
is required to liberate the desired enantiomeric form.
Alternatively, the specific stereoisomers may be synthesized by
using an optically active starting material, by asymmetric
synthesis using optically active reagents, substrates, catalysts or
solvents, or by converting one stereoisomer into the other by
asymmetric transformation.
[0055] Certain compounds of formula (1) may exist in different
stable conformational forms which may be separable. Torsional
asymmetry due to restricted rotation about an asymmetric single
bond, for example because of steric hindrance or ring strain, may
permit separation of different conformers. The compounds of the
present invention further include each conformational isomer of
compounds of formula (1) and mixtures thereof.
[0056] It follows that a single compound may exhibit more than one
type of isomerism. Included within the scope of the claimed
compounds present invention are all stereoisomers, geometric
isomers and tautomeric forms of the compounds of formula (I),
including compounds exhibiting more than one type of isomerism, and
mixtures of one or more thereof. Also included are acid addition or
base salts wherein the counterion is optically active, for example,
D-lactate or L-lysine, or racemic, for example, DL-tartrate or
DL-arginine.
[0057] The compounds of the present invention, and the salts
thereof, may exist in unsolvated as well as solvated forms with
pharmaceutically acceptable solvents such as water, ethanol, and
the like. The term `solvate` is used herein to describe a molecular
complex comprising the compound of the invention and one or more
pharmaceutically acceptable solvent molecules, for example,
ethanol. The term `hydrate` is employed when said solvent is
water.
[0058] Pharmaceutically acceptable solvates include hydrates and
other solvates wherein the solvent of crystallization may be
isotopically substituted, e.g. D.sub.2O, d.sub.6-acetone,
d.sub.6-DMSO. The compounds of the present invention may exist as
clathrates or other complexes.
[0059] Certain compounds of formula (1) and their salts and
solvates may exist in more than one crystal form. Polymorphs of
compounds represented by formula (1) form part of this invention
and may be prepared by crystallization of a compound of formula (1)
under different conditions. For example, using different solvents
or different solvent mixtures for recrystallization;
crystallization at different temperatures; various modes of
cooling, ranging from very fast to very slow cooling during
crystallization. Polymorphs may also be obtained by heating or
melting a compound of formula (1) followed by gradual or fast
cooling. The presence of polymorphs may be determined by solid
probe NMR spectroscopy, IR spectroscopy, differential scanning
calorimetry, powder X-ray diffraction or such other techniques.
[0060] This invention also includes isotopically-labeled compounds,
which are identical to those described by formula (2), but for the
fact that one or more atoms are replaced by an atom having an
atomic mass or mass number different from the atomic mass or mass
number usually found in nature. Examples of isotopes that can be
incorporated into compounds of the invention include isotopes of
hydrogen, carbon, nitrogen, oxygen, sulfur and fluorine, such as
.sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O, .sup.17O,
.sup.35S, .sup.36Cl, .sup.125I, .sup.129I, and .sup.18F
respectively. Compounds of the present invention, and
pharmaceutically acceptable salts of the compounds which contain
the aforementioned isotopes and/or other isotopes of other atoms
are within the scope of this invention. Certain
isotopically-labeled compounds of the present invention, for
example those into which radioactive isotopes such as .sup.3H and
.sup.14C are incorporated, are useful in drug and/or substrate
tissue distribution assays. Tritiated (i.e., .sup.3H), and
carbon-14 (i.e., .sup.14C), isotopes are particularly preferred for
their ease of preparation and detectability. Further, substitution
with heavier isotopes such as deuterium (i.e., .sup.2H), can afford
certain therapeutic advantages resulting from greater metabolic
stability, for example increased in vivo half-life or reduced
dosage requirements and, hence, may be preferred in some
circumstances. Isotopically labeled compounds of formula (2) of
this invention, and salts thereof can generally be prepared by
carrying out the procedures disclosed in the schemes and/or in the
Examples below, by substituting a readily available isotopically
labeled reagent for a non-isotopically labeled reagent.
[0061] Pharmaceutically acceptable salts, as used herein in
relation to compounds of the present invention, include
pharmaceutically acceptable inorganic and organic salts of said
compound. These salts can be prepared in situ during the final
isolation and purification of a compound, or by separately reacting
the compound or prodrug thereof, with a suitable organic or
inorganic acid and isolating the salt thus formed. Representative
salts include, but are not limited to, the acetate, aspartate,
benzoate, besylate, bicarbonate, dimesylate, diphosphate,
bisulfate, bitartrate, bromide, camsylate, carbonate, chloride,
citrate, edisylate, esylate, fumarate, gluceptate, hemifumarate,
hydrobromide, hydrochloride, isethionate, lactate, malate, maleate,
mesylate, napsylate, nitrate, oxalate, palmitate, pamoate,
phosphate, saccharate, stearate, succinate, sulfate, tartrate,
tosylate, and trifluoroacetate salts.
[0062] These may also include cations based on the alkali and
alkaline earth metals, such as sodium, lithium, potassium, calcium,
magnesium, and the like, as well as non-toxic ammonium, quaternary
ammonium, and amine cations including, but not limited to,
ammonium, tetramethylammonium, tetraethylammonium, methylamine,
dimethylamine, trimethylamine, triethylamine, ethylamine,
diethylamine and the like. For additional examples see, for
example, Berge, et al., J. Pharm. Sci., 66, 1-19 (1977) and
"Handbook of Pharmaceutical Salts: Properties, Selection, and Use"
by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).
[0063] The compounds of the present invention may be isolated and
used per se or in the form of their pharmaceutically acceptable
salts. In accordance with the present invention, compounds with
multiple basic nitrogen atoms can form salts with varying number of
equivalents ("eq.") of acid. It will be understood by practitioners
that all such salts are within the scope of the present
invention.
[0064] The present invention further includes prodrugs of compounds
of formula (1). A prodrug of a compound of formula (1) may be one
formed in a conventional manner with a functional group of the
compound, such as with an amino, hydroxy or carboxy group. The term
"prodrug" means a compound that is transformed in vivo to yield a
compound of formula (1) or a pharmaceutically acceptable salt of
the compound. The transformation may occur by various mechanisms,
such as through hydrolysis in blood. A discussion of the use of
prodrugs is provided by T. Higuchi and W. Stella, "Pro-drugs as
Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series,
and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987.
[0065] For example, as all of the compounds of the present
invention incorporate an amine functional group, a prodrug can be
formed by the replacement of a hydrogen atom in the amine group
with a group such as R-carbonyl, RO-carbonyl, NRR'-carbonyl where R
and R' are each independently (C.sub.1-C.sub.10)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, benzyl, or R-carbonyl is a natural
.alpha.-aminoacyl or natural .alpha.-aminoacyl-natural
.alpha.-aminoacyl, --C(OH)C(O)OY' wherein Y' is H,
(C.sub.1-C.sub.6)alkyl or benzyl, --C(OY.sub.0)Y.sub.1 wherein
Y.sub.0 is (C.sub.1-C.sub.4) alkyl and Y.sub.1 is
(C.sub.1-C.sub.6)alkyl, carboxy(C.sub.1-C.sub.6)alkyl,
amino(C.sub.1-C.sub.4)alkyl or mono-N-- or
di-N,N--(C.sub.1-C.sub.6)alkylaminoalkyl, --C(Y.sub.2)Y.sub.3
wherein Y.sub.2 is H or methyl and Y.sub.3 is mono-N-- or
di-N,N--(C.sub.1-C.sub.6)alkylamino, morpholino, piperidin-1-yl or
pyrrolidin-1-yl.
[0066] Similarly, if a compound of the present invention contains
an alcohol functional group, a prodrug can be formed by the
replacement of the hydrogen atom of the alcohol group with a group
such as (C.sub.1-C.sub.6)alkanoyloxy-methyl,
1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
(C.sub.1-C.sub.6)alkoxycarbonyloxymethyl,
N--(C.sub.1-C.sub.6)alkoxycarbonylaminomethyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkanoyl,
arylacyl and .alpha.-aminoacyl, or
.alpha.-aminoacyl-.alpha.-aminoacyl, where each .alpha.-aminoacyl
group is independently selected from the naturally occurring
L-amino acids, P(O)(OH).sub.2,
--P(O)(O)(C.sub.1-C.sub.6)alkyl).sub.2 or glycosyl (the radical
resulting from the removal of a hydroxyl group of the hemiacetal
form of a carbohydrate).
[0067] If a compound of the present invention contains a carboxylic
acid functional group, a prodrug can comprise an ester formed by
the replacement of the hydrogen atom of the acid group with a group
such as (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having
from 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having
from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to
6 carbon atoms, 1-(alkoxycarbonyl-oxy)ethyl having from 4 to 7
carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)-ethyl having from 5 to
8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9
carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10
carbon atoms, 3-phthalidyl, 4-crotonolactonyl,
gamma-butyrolacton-4-yl,
di-N,N--(C.sub.1-C.sub.2)alkylamino(C.sub.2-C.sub.3)alkyl (such as
.beta.-dimethylaminoethyl), carbamoyl-(C.sub.1-C.sub.2)alkyl,
N,N-di(C.sub.1-C.sub.2)alkylcarbamoyl-(C.sub.1-C.sub.2)alkyl and
piperidino-, pyrrolidino- or morpholino(C.sub.2-C.sub.3)alkyl.
Synthesis
[0068] In general, the compounds of formula (1) of this invention
may be prepared by methods that include processes known in the
chemical arts, particularly in light of the description contained
herein. Certain processes for the manufacture of the compounds of
formula (1) of this invention are illustrated by the following
reaction schemes. Other processes are described in the experimental
section. Some of the starting compounds for the reactions described
in the schemes and Examples are prepared as illustrated herein.
[0069] In Schemes 1, 3 and 4 below, generalized methods for
preparing the compounds of formula (1) are depicted.
##STR00004## ##STR00005##
[0070] In Scheme 1, aryl halide/sulfonates 1-1 can be prepared
beginning with commercially available starting materials and using
general synthetic techniques known to those skilled in the arts.
Additional support for the preparation of intermediates such as
1-1, wherein X is a halide or OSO.sub.2R, are provided in U.S. Pat.
No. 7,244,727, issued Jul. 17, 2007, the teachings of which are
incorporated herein, by reference, in their entirety. Compound 1-2
was prepared by the method of C. Palomo et al., Synthesis of
.beta.-Lactam Scaffolds for Ditopic Peptidomimetics, Organic
Letters (2007), 9(1), pages 101-104. Compounds 1-1 and 1-2 can be
coupled using a metal catalyst, such as palladium or copper to form
Compound 1-3. More specifically, 1-1 and 1-2 are heated to a
temperature between 80.degree. C. to 130.degree. C. in a solvent,
such as toluene, with cesium carbonate, palladium acetate and
2-dicyclohexyl phosphino-2',4',6'-triisopropylbiphenyl (X--PHOS)
under nitrogen for about 15-20 hours to form Compound 1-3.
[0071] Compound 1-3 is acylated with 1-4 using conditions and
reagents known to the skilled artisan (utilizing a mild base, such
as triethylamine or pyridine) to afford compound 1-5. Compound 1-4
is prepared by the method of Tarasov, Evgeniy V.; Henckens, Anja;
Ceulemans, Erik; Dehaen, Wim. A short total synthesis of cerpegin
by intramolecular hetero Diels-Alder cycloaddition reaction of an
acetylene tethered pyrimidine. Synlett (2000), (5), 625-626. The
protecting group (Pg) present in compound 1-5 can be removed by one
skilled in the art, utilizing conditions referenced in Greene's
Protective Groups in Organic Synthesis, 4.sup.th Ed., P. G. M Wuts
and T. W. Greene, Wiley-Interscience to afford compound 1-6. In the
case where Pg=t-butyldimethyl-silyl, deprotection can be
accomplished by a range of conditions including acidic and
fluoride-based conditions. Preferred conditions when Pg is
t-butyldimethylsilyl are dilute aqueous hydrochloric acid in
methanol at ambient temperature for 2-10 hours. Cyclization of
compound 1-6 to afford compound 1-7 can be accomplished utilizing a
wide range of basic conditions, including organic (e.g.
triethylamine) and inorganic (e.g. potassium carbonate) as the
bases, in an aprotic solvent at 20.degree. C. to 120.degree. C. to
provide the cyclic lactam structure 1-7. Preferred conditions for
this cyclization are triethylamine in acetonitrile at 40.degree. C.
to 120.degree. C. for 4-16 hours. Amination of compound 1-7 can be
accomplished with ammonia in a range of aprotic or protic solvents
at 0.degree. C. to 100.degree. C. for 4-20 hours. Preferred
conditions are ammonia in p-dioxane at ambient temperature for 4 to
24 hours.
[0072] Compounds of formula (1), which contain a carboxylic ester
functionality, can be hydrolyzed to the corresponding carboxylic
acid utilizing base or acid catalyzed hydrolysis conditions as
known in the art. A preferred method of hydrolysis is treating
compound (1) with aqueous lithium hydroxide in an organic solvent
at 20.degree. C. to 100.degree. C. for 1 to 10 hours.
##STR00006##
[0073] Compound 1B may be formed through a peptide coupling
reaction between Compound 1A and ammonia. Compound 1B may be used
to form Compound 1C by amide dehydration The dehydration may be
performed by treating the amide with SOCl.sub.2, POCl.sub.3,
PCl.sub.5, p-TosCl/pyridine, Tf.sub.2O/pyridine or with the
Vilsmeier reagent in combination with an organic or inorganic base.
The Vilsmeier reagent can be prepared by reacting dimethylformamide
(DMF) with oxalylchloride in acetonitrile, dichloromethane,
chloroform, dioxane, tetrahydrofuran (THF), or diethylether. In a
general procedure, the Vilsmeier reagent is formed in the desired
solvent for instance at a temperature between 0.degree. C. and room
temperature. The formation normally will be completed in 5-15
minutes. In a preferred embodiment a solution of the amide in the
desired solvent is added dropwise to the Vilsmeier reagent at a
temperature between 0.degree. C. and room temperature. The addition
normally will be completed in 10-20 minutes. For the formation of
the nitrile, two equivalents of a base are added. Preferably an
organic base, for instance pyridine or triethylamine (TEA) is used.
Inorganic bases may also be effective.
##STR00007##
[0074] In Scheme 2, the ketones (2-5) which are the equivalent of
compound 1-1 of Scheme 1, wherein m and n are each independently 1,
2 or 3, can be prepared utilizing the procedure of Zang, Y.;
Schuster, G., J. Org. Chem. 1994, 59, 1855-62. The monoprotected
diones (2-2) can be prepared using the procedures of Alonso, F.; et
al., Tetrahedron, 1995, 51(37), 10259-80, Crich, D.; Fortt, S.
Tetrahedron, 1989, 45(20), 6581-98, or Lee, S.; Fuchs, P., J. Amer.
Chem. Soc., 2002, 124, 13978-9.
##STR00008##
[0075] In Scheme 3, compound 1-5, and analogues thereto, may be
prepared from compounds 3-1 and 3-2 as described in the following
Preparation 4 and through methods known in the art.
##STR00009##
[0076] In Scheme 3A, compound 3-1, and analogues thereto, may be
prepared from compound 3A-1 and 3A-6 as described in the following
Preparation 4A and through methods known in the art.
[0077] Alternatively, in Scheme 4, compounds of formula (1) may
also be prepared from compound 4-7, which is prepared by the method
of Scheme 3 above, through methods known in the art.
##STR00010##
[0078] A pharmaceutical composition of the present invention
comprises a therapeutically effective amount of a compound of
formula (1) or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, vehicle, diluent or excipient.
A preferred pharmaceutical composition of the present invention
comprises a therapeutically effective amount of a compound of
formula (2), or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, vehicle, diluent or excipient.
Even more preferably, the pharmaceutical composition contains
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}-acetonitrile or a pharmaceutically acceptable
salt thereof,
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6-
(5H)-yl)-2-fluorophenyl)-cyclohexyl)acetic acid or a
pharmaceutically acceptable salt thereof,
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-oxazepin-6(5H)-
-yl)phenyl]-cyclohexyl}acetic acid or a pharmaceutically acceptable
salt thereof,
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-
-f][1,4]oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetonitrile or a
pharmaceutically acceptable salt thereof,
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]-
oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetic acid or a
pharmaceutically acceptable salt thereof, or
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydro-pyrimido[5,4-f][1,4]oxaze-
pin-6(5H)-yl)-2-fluorophenyl)-cyclohexyl)acetic acid or a
pharmaceutically acceptable salt thereof; and a pharmaceutically
acceptable carrier, vehicle, diluent or excipient.
[0079] The pharmaceutical compositions formed by combining the
compounds of this invention and the pharmaceutically acceptable
carriers, vehicles or diluents are then readily administered in a
variety of dosage forms such as tablets, powders, lozenges, syrups,
injectable solutions and the like. These pharmaceutical
compositions can, if desired, contain additional ingredients such
as flavorings, binders, excipients and the like.
[0080] Thus, for purposes of oral administration, tablets
containing various excipients such as sodium citrate, calcium
carbonate and/or calcium phosphate, may be employed along with
various disintegrants such as starch, alginic acid and/or certain
complex silicates, together with binding agents such as
polyvinylpyrrolidone, sucrose, gelatin and/or acacia. Additionally,
lubricating agents such as magnesium stearate, sodium lauryl
sulfate and talc are often useful for tabletting purposes. Solid
compositions of a similar type may also be employed as fillers in
soft and hard filled gelatin capsules. Preferred materials for this
include lactose or milk sugar and high molecular weight
polyethylene glycols. When aqueous suspensions or elixirs are
desired for oral administration, the active pharmaceutical agent
therein may be combined with various sweetening or flavoring
agents, coloring matter or dyes and, if desired, emulsifying or
suspending agents, together with diluents such as water, ethanol,
propylene glycol, glycerin and/or combinations thereof.
[0081] For parenteral administration, solutions of the compounds or
compositions of this invention in sesame or peanut oil, aqueous
propylene glycol, or in sterile aqueous solutions may be employed.
Such aqueous solutions should be suitably buffered if necessary and
the liquid diluent first rendered isotonic with sufficient saline
or glucose. These particular aqueous solutions are especially
suitable for intravenous, intramuscular, subcutaneous and
intraperitoneal administration. In this connection, the sterile
aqueous media employed are all readily available by standard
techniques known to those skilled in the art.
[0082] For intranasal administration or administration by
inhalation, the compounds or compositions of the invention are
conveniently delivered in the form of a solution or suspension from
a pump spray container that is squeezed or pumped by the patient or
as an aerosol spray presentation from a pressurized container or a
nebulizer, with the use of a suitable propellant, e.g.,
dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In
the case of a pressurized aerosol, the dosage unit may be
determined by providing a valve to deliver a metered amount. The
pressurized container or nebulizer may contain a solution or
suspension of a compound of this invention.
[0083] Capsules and cartridges (made, for example, from gelatin)
for use in an inhaler or insufflator may be formulated containing a
powder mix of a compound or compounds of the invention and a
suitable powder base such as lactose or starch.
[0084] Methods of preparing various pharmaceutical compositions
with a certain amount of active ingredient are known, or will be
apparent in light of this disclosure, to those skilled in this art.
For examples of methods of preparing pharmaceutical compositions,
see Remington's Pharmaceutical Sciences, Mack Publishing Company,
Easton, Pa., 19th Edition (1995).
[0085] In another aspect, the invention is directed to a
pharmaceutical composition, which comprises a therapeutically
effective amount of a first compound of formula (1), or a
pharmaceutically acceptable salt of the compound; a second compound
that is an antidiabetic agent selected from insulin and insulin
analogs; insulinotropin; biguanides; .alpha..sub.2-antagonists and
imidazolines; glitazones; aldose reductase inhibitors; glycogen
phosphorylase inhibitors; sorbitol dehydrogenase inhibitors; fatty
acid oxidation inhibitors; .alpha.-glucosidase inhibitors;
.beta.-agonists; phosphodiesterase inhibitors; lipid-lowering
agents; antiobesity agents; vanadate and vanadium complexes and
peroxovanadium complexes; amylin antagonists; glucagon antagonists;
growth hormone secretagogues; gluconeogenesis inhibitors;
somatostatin analogs; antilipolytic agents; a prodrug of the
antidiabetic agents, or a pharmaceutically acceptable salt of the
antidiabetic agents and the prodrugs.
[0086] In another aspect, the invention is directed to a kit
comprising: a first dosage form comprising a compound of formula
(1), or a pharmaceutically acceptable salt of the compound; and a
second dosage form comprising an antidiabetic agent selected from
insulin and insulin analogs; insulinotropin; biguanides;
.alpha..sub.2-antagonists and imidazolines; glitazones; aldose
reductase inhibitors; glycogen phosphorylase inhibitors; sorbitol
dehydrogenase inhibitors; fatty acid oxidation inhibitors;
.alpha.-glucosidase inhibitors; .beta.-agonists; phosphodiesterase
inhibitors; lipid-lowering agents; antiobesity agents; vanadate and
vanadium complexes and peroxovanadium complexes; amylin
antagonists; glucagon antagonists; growth hormone secretagogues;
gluconeogenesis inhibitors; somatostatin analogs; antilipolytic
agents; prodrugs of the antidiabetic agents, or a pharmaceutically
acceptable salts of the antidiabetic agents and the prodrug; and a
container for containing said first dosage (a) and said second
dosage (b). In a preferred embodiment of the kit, both the first
and the second dosage forms independently comprise a
pharmaceutically acceptable carrier or diluent.
[0087] In another aspect, the invention is directed to a method of
inhibiting DGAT-I comprising administering to a mammal an
inhibitory amount of a compound of formula (1), or a
pharmaceutically acceptable salt of the compound, either alone or
in combination with an antidiabetic agent as described above.
[0088] In another aspect, the invention is directed to a method of
treating a condition mediated by DGAT-1 inhibition comprising
administering to a mammal in need of such treatment a
therapeutically effective amount of a compound of formula (1), or a
pharmaceutically acceptable salt of the compound, either alone or
in combination with an antidiabetic agent as described above.
[0089] In the present invention, typically, the condition treated
is Type 2 diabetes, insulin resistance syndrome or obesity.
Preferably, the condition is treated by administering a compound of
formula (2), or a pharmaceutically acceptable salt thereof. More
preferably, the condition is treated by administering one of the
following compounds, or pharmaceutically acceptable salts thereof:
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}-acetonitrile,
2-(4-(4-(4-amino-5-oxo-7,8-dihydro-pyrimido[5,4-f][1,4]oxazepin-6(5H)-yl)-
-2-fluorophenyl)-cyclohexyl)acetic acid,
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-oxazepin-6(5H)-
-yl)phenyl]-cyclohexyl}acetic acid,
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl}cyclohexyl)acetonitrile,
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]-
oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetic acid, or
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydro-pyrimido[5,4-f][1,4]oxaze-
pin-6(5H)-yl)-2-fluorophenyl)-cyclohexyl)acetic acid.
[0090] Preferably, the mammal treated is a human.
[0091] In an alternate embodiment, the condition treated is
impaired glucose tolerance, hyperglycemia, diabetic complications
such as sugar cataracts, diabetic neuropathy, diabetic nephropathy,
diabetic retinopathy and diabetic cardiomyopathy, anorexia nervosa,
bulimia, cachexia, hyperuricemia, hyperinsulinemia,
hypercholesterolemia, hyperlipidemia, dyslipidemia, mixed
dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver
disease, atherosclerosis, arteriosclerosis, acute heart failure,
congestive heart failure, coronary artery disease, cardiomyopathy,
myocardial infarction, angina pectoris, hypertension, hypotension,
stroke, ischemia, ischemic reperfusion injury, aneurysm,
restenosis, vascular stenosis, solid tumors, skin cancer, melanoma,
lymphoma, breast cancer, lung cancer, colorectal cancer, stomach
cancer, esophageal cancer, pancreatic cancer, prostate cancer,
kidney cancer, liver cancer, bladder cancer, cervical cancer,
uterine cancer, testicular cancer and ovarian cancer.
[0092] The present invention also relates to therapeutic methods
for treating the above described conditions in a mammal, including
a human, wherein a compound of formula (1) of this invention is
administered as part of an appropriate dosage regimen designed to
obtain the benefits of the therapy. The appropriate dosage regimen,
the amount of each dose administered and the intervals between
doses of the compound will depend upon the compound of formula (1)
of this invention being used, the type of pharmaceutical
compositions being used, the characteristics of the subject being
treated and the severity of the conditions.
[0093] In general, an effective dosage for the compounds of the
present invention is in the range of 0.01 mg/kg/day to 30
mg/kg/day, preferably 0.01 mg/kg/day to 5 mg/kg/day of active
compound in single or divided doses. Some variation in dosage will
necessarily occur, however, depending on the condition of the
subject being treated. The individual responsible for dosing will,
in any event, determine the appropriate dose for the individual
subject. Practitioners will appreciate that "kg" refers to the
weight of the patient measured in kilograms.
[0094] The compounds or compositions of this invention may be
administered in single (e.g., once daily) or multiple doses or via
constant infusion. The compounds of this invention may also be
administered alone or in combination with pharmaceutically
acceptable carriers, vehicles or diluents, in either single or
multiple doses. Suitable pharmaceutical carriers, vehicles and
diluents include inert solid diluents or fillers, sterile aqueous
solutions and various organic solvents.
[0095] The compounds or compositions of the present invention may
be administered to a subject in need of treatment by a variety of
conventional routes of administration, including orally and
parenterally, (e.g., intravenously, subcutaneously or
intramedullary). Further, the pharmaceutical compositions of this
invention may be administered intranasally, as a suppository, or
using a "flash" formulation, i.e., allowing the medication to
dissolve in the mouth without the need to use water.
Exemplification
[0096] The Examples set forth herein below are for illustrative
purposes only. The compositions, methods, and various parameters
reflected herein are intended only to exemplify various aspects and
embodiments of the invention, and are not intended to limit the
scope of the claimed invention in any way.
[0097] The abbreviations used in the exemplification are defined as
follows: [0098] DCM is dichloromethane, [0099] DMF is
dimethylformamide, [0100] DPPF is
1,1'-Bis(diphenylphosphino)ferrocene, [0101] EDCl is
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, [0102] EtOAc is
ethyl acetate, [0103] HCl is hydrochloric acid, [0104] HOBt is
N-hydroxybenzotriazole, [0105] MTHF is 2-Methyltetrahydrofuran,
[0106] RT is retention time in minutes, [0107] TEA is
triethylamine, [0108] TFA is trifluoroacetic acid, [0109] Tf.sub.2O
is Trifluoromethanesulfonic(triflic)anhydride, [0110] THF is
tetrahydrofuran, and [0111] X--PHOS is 2-dicyclohexyl
phosphino-2',4',6'-triisopropylbiphenyl.
[0112] Unless noted otherwise, all reactants were obtained
commercially.
[0113] Unless otherwise noted, purifications were performed using a
Biotage.RTM. SNAP FLASH purification cartridge (silica gel).
[0114] All Combiflash.RTM. purifications, discussed herein, were
performed using a CombiFlash.RTM. Companion system (Teledyne Isco;
Lincoln, Nebr.) utilizing packed RediSep.RTM. silica columns.
[0115] Mass Spectra were recorded on a Waters.RTM. (Waters Corp.;
Milford, Mass.) Micromass.RTM. Platform II spectrometer. Unless
otherwise specified, mass spectra were recorded on a Waters.RTM.
(Milford, Mass.) Micromass.RTM. Platform II spectrometer.
[0116] Proton NMR chemical shifts are given in parts per million
downfield from tetramethylsilane and were recorded on a Varian
Unity 400 MHz (megaHertz) spectrometer (Varian Inc.; Palo Alto,
Calif.).
[0117] The compounds of the present invention, described in
Examples 1-48, were prepared, or subsequently derived from a
compound prepared, by the method of Scheme I using the compounds of
Preparations 1, 2, 3 or 4.
Preparation 1
Methyl{trans-4-[4-(4-chloro-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin--
6-(5H)-yl)phenyl]cyclohexyl}acetate
##STR00011##
[0118]
Methyl{trans-4-[4-(4-chloro-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-o-
xazepin-6-(5H)-yl)phenyl]cyclohexyl}acetate was prepared as
follows
##STR00012##
[0120] Step 1.
Methyl(trans-4-{4-[(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)amino]-phenyl-
}cyclohexyl)acetate, shown above, was prepared as follows.
##STR00013##
[0121]
Methyl[trans-4-[4-[[(trifluoromethyl)sulfonyl]oxy]phenyl]cyclohexyl-
]acetate, shown above, which is identified as Compound 55 in U.S.
Pat. No. 7,244,727, issued Jul. 17, 2007, was prepared according to
the method described therein. A mixture of methyl
[trans-4-[4-[[(trifluoromethyl)-sulfonyl]oxy]phenyl]cyclohexyl]acetate
(10.1 g, 26.6 mmol), 2-{[tert-butyl(dimethyl)silyl]oxy}ethanamine
(5.59 g, 31.9 mmol), which can be prepared by various methods
including those disclosed in Journal of the American Chemical
Society, 129(37), 11408-11420; 2007, Organic Letters, 9(1),
101-104; 2007 or Bioorganic & Medicinal Chemistry, 13(11),
3821-3839; 2005, cesium carbonate (8.65 g, 26.6 mmol), palladium
acetate (0.60 g, 2.66 mmol) and X--PHOS (1.27 g, 2.66 mmol) in
toluene (53 mL) under nitrogen was heated in a sealed tube at
120.degree. C. for 16 hours. The reaction was cooled, diluted into
EtOAc, washed with water (2.times.), saturated aqueous brine, dried
over sodium sulfate and concentrated to afford a dark oil.
Chromatography (330 g Biotage Snap Cartridge.RTM. silica gel
column, 0-15% EtOAc:heptane) afforded methyl
(trans-4-{4-[(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)-amino]phenyl}cyclo-
hexyl)acetate as a light-yellow oil, 6.70 g. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 7.02, (d, 2H), 6.61 (d, 2H), 3.80 (m, 2H),
3.64 (s, 3H), 3.20 (m, 2H), 2.37 (m, 1H), 2.24 (m, 2H), 1.85 (m,
5H), 1.44 (m, 2H), 1.13 (m, 2H), 0.87 (s, 9H), 0.04 (s, 6H).
m/z=406.4 (M+1).
[0122] Step 2. Methyl
[trans-4-(4-{(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)[(4,6-dichloropyrim-
idin-5-yl)carbonyl]amino}phenyl)cyclohexyl]acetate, shown below,
was prepared as follows.
##STR00014##
[0123] To a stirred, cooled (0.degree. C.) solution of methyl
(trans-4-{4-[(2-{[tert-butyl-(dimethyl)silyl]oxy}ethyl)amino]phenyl}cyclo-
hexyl)acetate (9.7 g, 24.0 mmol), from Step 1, and TEA (3.53 mL,
25.3 mmol) in THF (60 mL) was added dropwise a solution of
4,6-dichloropyrimidine-5-carbonyl chloride (5.31 g, 25.1 mmol) in
THF (20 mL). After 2 hours, the reaction was concentrated in vacuo,
diluted into EtOAc, washed with water (3.times.), saturated aqueous
brine, dried over sodium sulfate and concentrated in vacuo, to
afford an oil, which was carried on to the next step without
further purification. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
8.57 (s, 1H), 7.35 (d, 2H), 7.03 (d, 2H), 4.00 (m, 2H), 3.87 (m,
2H), 3.63 (s, 3H), 2.37 (m, 1H), 2.22 (m, 2H), 1.82 (m, 5H), 1.36
(m, 2H), 1.11 (m, 2H), 0.83 (s, 9H), 0.02 (s, 6H). m/z=580.3
(M+1).
[0124] Step 3.
Methyl(trans-4-{4-[[4,6-dichloropyrimidin-5-yl)carbonyl](2-hydroxyethyl)a-
mino]phenyl}cyclohexyl)acetate, shown below, was prepared as
follows.
##STR00015##
[0125] A solution of methyl
[trans-4-(4-{(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)[(4,6-dichloropyrim-
idin-5-yl)carbonyl]amino}phenyl)cyclohexyl]acetate (14.0 g, 24.0
mmol), from Step 2, in a methanolic solution of HCl (3 mL of
concentrated aqueous HCl in 97 mL of methanol) was stirred at room
temperature for 30 minutes. Methanol was removed in vacuo, the
residue was dissolved in EtOAc, washed with saturated aqueous
sodium bicarbonate, saturated aqueous brine, dried over sodium
sulfate and concentrated in vacuo to afford an oil, which was
carried on to the next step without further purification. .sup.1H
NMR (400 MHz, CDCl.sub.3): .delta. 8.59 (s, 1H), 7.32 (d, 2H), 7.04
(d, 2H), 4.08 (m, 2H), 3.92 (m, 2H), 3.63 (s, 3H), 2.38 (m, 1H),
2.23 (m, 2H), 1.82 (m, 5H), 1.39 (m, 2H), 1.11 (m, 2H). m/z=466.2
(M+1).
[0126] Step 4. The title compound of Preparation 1 was prepared as
follows. A slurry of methyl
(trans-4-{4-[[4,6-dichloropyrimidin-5-yl)carbonyl](2-hydroxy-ethyl)amino]-
phenyl}cyclohexyl)acetate (4.78 g, 10.2 mmol, unpurified material
from Step 3) and TEA (4.15 g, 41 mmol) in acetonitrile was stirred
at 80.degree. C. for 6 hours. The reaction was cooled, concentrated
in vacuo, diluted into EtOAc, washed with water (3.times.),
saturated aqueous brine, dried over sodium sulfate and concentrated
in vacuo to afford a yellow solid. This material was slurried in
methanol (10 mL), filtered, the solids washed with methanol
(2.times.3 mL) and dried in vacuo to afford the title compound as a
yellow solid, 4.03 g. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
8.75 (s, 1H), 7.22 (s, 4H), 4.75 (m, 2H), 4.03 (m, 2H), 3.63 (s,
3H), 2.50 (m, 1H), 2.23 (m, 2H), 1.87 (m, 5H), 1.44 (m, 2H), 1.19
(m, 2H). m/z=430.3 (M+1).
Preparation 2
Methyl
2-((1S,4s)-4-(4-((R)-4-chloro-8-methyl-5-oxo-7,8-dihydropyrimido-[5-
,4-f][1,4]oxazepin-6(5H)-yl)phenyl)cyclohexyl)acetate
##STR00016##
[0128]
Methyl(trans)-4-(4-((R)4-chloro-8-methyl-5-oxo-7,8-dihydro-pyrimido-
[5,4-f][1,4]oxazepin-6(5H)-yl)phenyl]cyclohexyl}acetate was
Prepared as follows.
[0129] Step 1. Methyl
[trans-4-(4-{[(2R)-2-{[tert-butyl(dimethyl)silyl]oxy}propyl]-amino}phenyl-
)cyclohexyl]acetate, shown below, was prepared as follows.
##STR00017##
[0130] A mixture of methyl
[trans-4-[4-[[(trifluoromethyl)sulfonyl]oxy]phenyl]-cyclohexyl]acetate
(5.00 g, 13.1 mmol), shown below,
##STR00018##
(R)-2-(tert-butyldimethylsilyloxy)propan-1-amine (2.99 g, 15.8
mmol), shown below,
##STR00019##
which can be prepared by various methods including those disclosed
in the Journal of Organic Chemistry, 72(20), 7726-7735; 2007,
cesium carbonate (5.14 g, 15.8 mmol), palladium acetate (310 mg,
1.32 mmol) and X--PHOS (627 mg, 1.32 mmol) in toluene (100 mL)
under nitrogen was heated in a sealed tube at 120.degree. C. for 16
hours. The reaction was cooled, diluted into EtOAc (500 mL), washed
with water (2.times.200 mL), saturated aqueous brine, dried over
sodium sulfate and concentrated to afford a dark oil.
Chromatography (120 g silica gel column, 3-15% EtOAc:heptane)
afforded the methyl
[trans-4-(4-{[(2R)-2-{[tert-butyl(dimethyl)silyl)oxy}propyl]amino}phenyl)-
cyclohexyl]acetate as a light-yellow oil, 4.55 g (86%). m/z=420.1
(M+1).
[0131] Step 2.
Methyl[trans-4-(4-{[(2R)-2-{[tert-butyl(dimethyl)silyl]oxy}propyl]-[(4,6--
dichloropyrimidin-5-yl)carbonyl]amino}phenyl)cyclohexyl]acetate,
shown below, was prepared as follows.
##STR00020##
[0132] A mixture of 4,6-dichloropyrimidine-5-carbonyl chloride
(2.27 g, 10.7 mmol), methyl
[trans-4-(4-{[(2R)-2-{[tert-butyl(dimethyl)silyl]oxy}propyl]-amino}phenyl-
)cyclohexyl]acetate (4.50 g, 10.7 mmol), from Step 1, and TEA (2.24
mL, 16.1 mmol) in THF (150 mL) was stirred at room temperature
under nitrogen for 14 hours. The reaction mixture was concentrated
to remove THF. The residue was diluted with EtOAc (300 mL), washed
with water (2.times.200 mL), dried over MgSO.sub.4 and
concentrated. The crude material was purified by a 120 g silica gel
column eluted with 3-15% EtOAc in heptane to give methyl
[trans-4-(4-{[(2R)-2-{[tert-butyl(dimethyl)silyl]oxy}propyl][(4,6-dichlor-
opyrimidin-5-yl)carbonyl]amino}phenyl)cyclohexyl]acetate as a
colorless oil 4.01 g (63%). m/z=594.2 (M+1). 1H NMR (400 MHz,
chloroform-d) .delta. -0.06 (s, 6H) 0.71 (s, 9H) 0.99-1.14 (m, 2H)
1.25-1.30 (m, 3H) 1.30-1.42 (m, 2H) 1.78 (dd, J=28.30, 11.90 Hz,
5H) 2.20 (d, J=7.03 Hz, 2H) 2.28-2.39 (m, 1H) 3.64 (s, 3H)
3.83-3.97 (m, 2H) 4.04-4.14 (m, 1H) 7.00 (d, J=8.20 Hz, 2H) 7.19
(d, J=8.59 Hz, 2H) 8.53 (s, 1H).
[0133] Step 3. The title compound of Preparation 2 was prepared as
follows. 4M HCl in dioxane (25 mL) was added to a solution of
methyl
[trans-4-(4-{[(2R)-2-{[tert-butyl(dimethyl)silyl]oxy}propyl][(4,6-dichlor-
opyrimidin-5-yl)carbonyl]amino}phenyl)cyclohexyl]acetate (3.95 g,
6.72 mmol), from Step 2, in methanol (50 mL). The mixture was
stirred at 23.degree. C. for 30 minutes. The reaction mixture was
concentrated to remove the solvent. The residue was dissolved in
acetonitrile (200 mL), then K.sub.2CO.sub.3 (1.86 g, 13.5 mmol) and
5 Angstrom molecular sieves (1.0 g) were added to it. The reaction
mixture was stirred at 80.degree. C. for 30 hours. EtOAc (250 mL)
and water (250 mL) were added to reaction mixture. The organic
layer was separated and dried over MgSO.sub.4 and concentrated. The
crude material was purified by a 120 g silica gel column eluted
with 30-50% EtOAc in heptane to give a colorless oil 1.85 g (61%)
as the title compound. m/z=444.1 (M+1). 1H NMR (400 MHz,
chloroform-d) .delta. 1.09-1.23 (m, 2H) 1.43 (d, J=6.64 Hz, 3H)
1.44-1.57 (m, 2H) 1.80-1.96 (m, 5H) 2.26 (d, J=7.05 Hz, 2H)
2.44-2.55 (m, 1H) 3.68 (s, 3H) 3.80-3.95 (m, 2H) 5.00-5.12 (m, 1H)
7.29 (s, 4H) 8.76 (s, 1H).
Preparation 3
Methyl(trans-4-{4-[(7S)-4-chloro-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f]-
[1,4]oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetate
##STR00021##
[0135] The compound
methyl(trans-4-{4-[(7S)-4-chloro-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f-
][1,4]oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetate was prepared as
follows.
[0136] Step 1. (S)-1-(Tert-butyldimethylsilyloxy)propan-2-amine,
shown below, was prepared as follows.
##STR00022##
[0137] Tert-butyldimethylsilyl chloride (22.1 g, 146 mmol) was
added to a stirred solution of the (S)-2-aminopropan-1-ol (10 g,
130 mmol) and TEA (23.9 ml, 172 mmol) in 325 ml of dichloromethane.
The heterogeneous mixture was stirred at room temperature for 18
hours. The reaction was diluted into EtOAc and the organic layer
was washed with brine then dried over sodium sulfate and
concentrated in vacuo to recover 20.37 g of product. 1H NMR (400
MHz, chloroform-d) .delta. 0.05 (s, 6H) 0.89 (s, 9H) 1.01 (d,
J=6.64 Hz, 3H) 2.87-3.07 (m, 1H) 3.18-3.35 (m, 1H) 3.51 (dd,
J=9.75, 4.35 Hz, 1H) m/z=190.2 (M+1).
[0138] Step 2.
2-((1S,4r)-4-(4-((S)-1-(tert-butyldimethylsilyloxy)propan-2-ylamino)pheny-
l)cyclohexyl)acetic acid, shown below, was prepared as follows
##STR00023##
[0139] A mixture of
methyl[trans-4-(4-{[(trifluoromethyl)sulfonyl]oxy}-phenyl)cyclohexyl]acet-
ate (1 g, 3 mmol),
(S)-1-(tert-butyldimethylsilyloxy)-propan-2-amine (0.597 g, 3.16
mmol), palladium(II) acetate (0.061 g, 0.271 mmol), X--PHOS (0.125
g, 0.263 mmol), cesium carbonate (1.03 g, 3.16 mmol) were diluted
in 26.3 ml of toluene. The mixture was purged with nitrogen flow
for a few minutes and then capped tightly. The reaction was then
heated to 120.degree. C. with stirring for 16 hours. The reaction
was subsequently cooled, and diluted with EtOAc. The organic
mixture was washed with water and brine, dried over MgSO.sub.4 and
concentrated. The product was columned on silica from 5-20% EtOAc
in heptane to recover 0.82 g of methyl trans
2S-(4-(4-(1-(tert-butyldimethylsilyloxy)propan-2-ylamino)phenyl)cyclohexy-
l)acetate. 1H NMR (400 MHz, chloroform-d) .delta. 0.03 (d, J=4.98
Hz, 6H) 0.89 (s, 9H) 1.05-1.19 (m, 2H) 1.18 (d, J=6.22 Hz, 3H) 1.44
(q, J=12.44 Hz, 2H) 1.76-1.93 (m, 5H) 2.23 (d, J=7.05 Hz, 2H) 2.34
(t, J=12.03 Hz, 1H) 3.42-3.65 (m, 3H) 3.67 (s, 3H) 6.55 (d, J=8.29
Hz, 2H) 6.99 (d, J=8.29 Hz, 2H). m/z=420.3 (M+1)
[0140] Step 3.
Methyl[trans-4-(4-{[(3S)-2-{[tert-butyl(dimethyl)silyl]oxy}propyl]-[(4,6--
dichloropyrimidin-5-yl)carbonyl]amino}phenyl)cyclohexyl]acetate,
shown below, was prepared as follows:
##STR00024##
[0141] 4,6-Dichloropyrimidine-5-carbonyl chloride (0.454 g, 2.15
mmol) was added to a solution of methyl trans
2S-(4-(4-(1-(tert-butyldimethylsilyloxy)propan-2-ylamino)phenyl)cyclohexy-
l)acetate (0.82 g, 2.0 mmol), from Step 2, and diispropylethylamine
(0.418 ml, 2.4 mmol) in 4.16 ml of MTHF at 0.degree. C. and stirred
cold for 1.5 hours. Added water and EtOAc; separated layers. Washed
organic layer with Saturated sodium bicarbonate, brine then dried
and concentrated. Columned product on silica from 10-30% EtOAc in
heptane to recover 0.95 g of product. 1H NMR (400 MHz,
chloroform-d) .delta. 0.06 (d, J=4.98 Hz, 6H) 0.90 (s, 9H)
0.99-1.18 (m, 2H) 1.17 (d, J=6.64 Hz, 3H) 1.28-1.45 (m, 2H)
1.73-1.94 (m, 5H) 2.23 (d, J=6.64 Hz, 2H) 2.38 (t, J=11.61 Hz, 1H)
3.51-3.66 (m, 2H) 3.67 (s, 3H) 4.84-5.03 (m, 1H) 7.03 (d, J=8.71
Hz, 2H) 7.34 (br. s., 2H) 8.50 (s, 1H). m/z=597.2 (M+4)
[0142] Step 4.
Methyl[trans-4-(4-{[(3S)-2-{oxy}propyl][(4,6-dichloropyrimidin-5-yl)carbo-
nyl]amino}phenyl)cyclohexyl]acetate, shown below, was prepared as
follows.
##STR00025##
[0143]
Methyl[trans-4-(4-{[(3S)-2-{[tert-butyl(dimethyl)silyl]oxy}propyl][-
(4,6-dichloropyrimidin-5-yl)carbonyl]amino}phenyl)cyclohexyl]acetate
(0.95 g, 1.6 mmol), from Step 3, and 7.4 ml of 1% HCl-methanol (3
mL of 36% aqueous HCl and 97 mL of methanol) were stirred at room
temperature and monitored by LC-MS. The reaction mixture was
concentrated to remove methanol. The crude material was dissolved
into EtOAc (200 ml). The EtOAc solution was washed with NaHCO.sub.3
aqueous solution (2.times.100 ml), water (100 ml) and brine (100
mL), dried over MgSO.sub.4 and concentrated to recover 0.77 g of
product. 1H NMR (400 MHz, chloroform-d) .delta. 1.03-1.17 (m, 2H)
1.19 (d, J=7.05 Hz, 3H) 1.29-1.46 (m, 2H) 1.69-1.91 (m, 5H) 2.23
(d, J=7.05 Hz, 2H) 2.38 (t, J=12.23 Hz, 1H) 3.60 (dd, J=11.61, 9.12
Hz, 1H) 3.67 (s, 3H) 3.78 (dd, J=11.82, 4.35 Hz, 1H) 4.85-5.04 (m,
1H) 7.07 (d, J=8.71 Hz, 2H) 7.33 (br. s., 2H) 8.52 (s, 1H)
m/z=483.1 (M+4).
[0144] Step 5. The title compound of Preparation 3 was prepared as
follows.
Methyl[trans-4-(4-{[(3S)-2-{oxy}propyl][(4,6-dichloropyrimidin-5-
-yl)carbonyl]-amino}phenyl)cyclohexyl]acetate (0.8 g, 1.66 mmol),
potassium carbonate (0.46 g, 3.33 mmol) and 200 mg of molecular
sieves were mixed into 41.6 ml of acetonitrile. The mixture was
added to sealed tube and heated at 80.degree. C. overnight. The
reaction mixture was concentrated and then diluted with EtOAc and
water. Separated layers and washed organic layer with brine, then
dried and concentrated. Columned product on silica in 40% ethyl
acetate in heptane to recover 0.22 g. 1H NMR (400 MHz,
chloroform-d) .delta. 1.06 (d, J=7.05 Hz, 3H) 1.10-1.25 (m, 2H)
1.42-1.55 (m, 2H) 1.73-1.98 (m, 5H) 2.26 (d, J=6.64 Hz, 2H)
2.41-2.59 (m, 1H) 3.69 (s, 3H) 4.27-4.42 (m, 1H) 4.45-4.70 (m, 2H)
7.17 (d, J=8.29 Hz, 2H) 7.30 (d, J=8.71 Hz, 2H) 8.76 (s, 1H)
m/z=444.0 (M+1).
Preparation 4
Methyl
2-(4-(4-(4-chloro-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5-
H)-yl)-3-fluorophenyl)cyclohexyl)acetate
##STR00026##
[0146] Methyl
2-(4-(4-(4-chloro-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-yl)-
-3-fluorophenyl)cyclohexyl)acetate, shown above, was prepared as
follows.
[0147] Step 1. 1-(Bromomethyl)benzene (4.92 g, 28.8 mmol) and
K.sub.2CO.sub.3 (3.98 g, 28.8 mmol) were added to a solution of
4-bromo-2-fluorophenol (5.0 g, 26 mmol) in acetone. The reaction
mixture was stirred at room temperature for 18 hours. Water (100
mL) and EtOAc (100 mL) were added, the organic layer was separated
and dried over MgSO.sub.4 and concentrated to give
1-(benzyloxy)-4-bromo-2-fluorobenzene as a yellow solid 7.06 g.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.24-7.41 (m, 5H), 7.20
(m, 1H), 7.16 (m, 1H), 6.82 (m, 1H), 5.10 (m, 2H).
[0148] Step 2. Methyl
2-(4-(4-(benzyloxy)-3-fluorophenyl)cyclohex-3-enyl)acetate, shown
below, was prepared as follows.
##STR00027##
[0149] Pd(PPh.sub.3).sub.4 (16.2 mg, 0.014 mmol) and CsCO.sub.3
(142 mg, 0.428 mmol) were added to a solution of
1-(benzyloxy)-4-bromo-2-fluorobenzene (130 mg, 0.464 mmol) and
methyl
2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)acetate
(100 mg, 0.357 mmol), shown below, and prepared as described in
Preparation 4A,
##STR00028##
in THF (2 mL) under nitrogen.
[0150] The reaction mixture was refluxed with stirring for 24
hours. LC-MS showed desired product. The reaction mixture was
cooled, diluted with EtOAc (30 mL), washed with water and brine,
dried over MgSO.sub.4 and concentrated. The crude material was
purified by chromatography (12 g silica gel column eluted with
5-10% EtOAc:heptane) to give methyl
2-(4-(4-(benzyloxy)-3-fluorophenyl)cyclohex-3-enyl)acetate as a
white solid (110 mg). .sup.1H NMR (400MHz, CDCl.sub.3): .delta.
7.28-7.42 (m, 5H), 7.10 (dd, 1H), 7.0 (d, 1H), 6.88 (t, 1H), 6.0
(m, 1H), 5.1 (s, 2H), 3.66 (s, 3H), 2.38 (m, 2H), 2.3 (d, 2H), 2.2
(m, 1H), 2.1 (m,1H), 1.9 (m,2H), 1.4 (m, 1H). m/z=355.4(M+1).
[0151] Step 3. Methyl
2-(4-(3-fluoro-4-hydroxyphenyl)cyclohexyl)acetate, shown below, was
prepared as follows.
##STR00029##
[0152] Methyl
2-(4-(4-(benzyloxy)-3-fluorophenyl)cyclohex-3-enyl)acetate (520 mg,
1.47 mmol) was dissolved in ethanol (15 mL) and EtOAc (15 mL). 20%
Pd(OH).sub.2/C (150 mg) was added to it. The reaction mixture was
shaken under 50 psi hydrogen for 20 hours. The mixture was
filtrated through diatomaceous earth (Celite.RTM.) to remove
catalyst and then concentrated. The purification was done by
chromatography (12 g RediSep.RTM. silica gel column (Teledyne ISCO,
Lincoln, Nebr.) with 5-10% EtOAc:heptane) to give methyl
2-(4-(4-(benzyloxy)-3-fluorophenyl)cyclohex-3-enyl)acetate as a
colorless oil (320 mg). .sup.1H NMR (400MHz, CDCl3): .delta.
6.80-7.0 (m, 3H), 5.58 (d, 1H), 3.63 (s, 3H), 2.24-2.5 (m, 1H),
2.4, 2.2 (d, 2H), 1.8 (m,3H), 1.6 (m, 4H), 1.4 (m, 1H), 1.12 (m,
1H). m/z=265.3 (M-1).
[0153] Step 4. Methyl
2-(4-(3-fluoro-4-(trifluoromethylsulfonyloxy)phenyl)cyclohexyl)acetate,
shown below, was prepared as follows.
##STR00030##
[0154] To a stirred solution of phenol (310 mg, 1.16 mmol) and TEA
(0.24 mL, 1.75 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added a triflic
anhydride solution (413 mg, 1.46 mmol) dropwise. The mixture was
stirred at 0.degree. C. for 2 hours. The reaction mixture was
washed with water (.times.2) and brine (.times.1), dried over
Na.sub.2SO.sub.4 and filtered. The organic solution was
concentrated and purified by chromatography (12 g silica gel column
with 3-15% EtOAc:heptane) to give methyl
2-(4-(3-fluoro-4-(trifluoromethylsulfonyloxy)phenyl)cyclohexyl)ace-
tate as a colorless oil (380 mg). m/z=397.4 (M-1).
[0155] Step 5. Methyl
2-(4-(4-(2-(tert-butyldimethylsilyloxy)ethylamino)-3-fluorophenyl)cyclohe-
xyl)acetate, shown below, was prepared as follows.
##STR00031##
[0156] A mixture of methyl
2-(4-(3-fluoro-4-(trifluoromethylsulfonyloxy)phenyl)cyclohexyl)acetate
(190 mg, 0.477 mmol), 2-{[tert-butyl(dimethyl)silyl]oxy}ethanamine
(100 mg, 0.572 mmol), cesium carbonate (171 mg, 0.525 mmol),
palladium acetate (11 mg, 0.048 mmol) and (X--PHOS) (23 mg, 0.048
mmol) in toluene (5 mL), under nitrogen, was heated in a sealed
tube at 120.degree. C. for 16 hours. The reaction was cooled,
diluted into EtOAc, washed with water (2.times.), saturated aqueous
brine, dried over sodium sulfate and concentrated to afford a dark
oil. Chromatography (12 g silica gel column, 3-15% EtOAc:heptane)
afforded methyl
2-(4-(4-(2-(tert-butyldimethyl-silyloxy)ethylamino)-3-fluorophenyl)-cyclo-
hexyl)acetate as a light-yellow oil, 110 mg. 1H NMR (400 MHz,
CDCl3); .delta. 7.2 (m, 1H), 6.95 (m, 1H), 6.8 (m, 1H), 3.8 (m,
2H), 3.64 (s, 3H), 3.2 (m, 2H), 2.5, 2.3 (m, 1H), 2.4, 2.2 (d, 2H),
1.85 (m, 2H), 1.64 (m, 5H), 1.4 (m, 1H), 1.1 (m, 1H), 0.88 (s, 9H),
0.06 (s, 6H). m/z=424.2 (M+1).
[0157] Step 6. Methyl
2-(4-(4-(N-(2-(tert-butyldimethylsilyloxy)ethyl)-4,6-dichloropyrimidine-5-
-carboxamido)-3-fluorophenyl)cyclohexyl)acetate, shown below, was
prepared as follows.
##STR00032##
[0158] A mixture of 4,6-dichloropyrimidine-5-carbonyl chloride
(82.5 mg, 0.390), methyl
2-(4-(4-(2-(tert-butyldimethylsilyloxy)ethylamino)-3-fluorophenyl)cyclohe-
xyl)acetate (110 mg, 0.26 mmol) and TEA (0.054 mL, 0.39 mmol) in
THF (2 mL) was stirred at room temperature under nitrogen for 14
hours. The reaction mixture was concentrated to remove THF. The
residue was diluted with EtOAc, washed with water, dried over
MgSO.sub.4 and concentrated. The crude material was purified by a
12 g silica gel column eluted with 3-15% EtOAc in heptane to give
methyl
2-(4-(4-(N-(2-(tert-butyldimethylsilyloxy)ethyl)-4,6-dichloropyrimidine-5-
-carboxamido)-3-fluorophenyl)cyclohexyl)acetate a colorless oil (60
mg). m/z=598.2 (M+1).
[0159] Step 7. The title compound of Preparation 4 was prepared as
follows. 4M HCl in dioxane (0.5 mL) was added to a solution of
methyl
2-(4-(4-(N-(2-(tert-butyldimethylsilyloxy)ethyl)-4,6-dichloropyrimidine-5-
-carboxamido)-3-fluorophenyl)cyclohexyl)acetate (30 mg, 0.05 mmol)
in methanol (1 mL). The mixture was stirred at 23.degree. C. for 30
minutes. The reaction mixture was concentrated to remove solvent.
The residue was dissolved in acetonitrile (1 mL) and NEt.sub.3
(0.05 mL) was added to it. The reaction mixture was stirred at
80.degree. C. for 18 hours. EtOAc (15 mL) and water (15 mL) were
added to reaction mixture. The organic layer was separated and
dried over MgSO.sub.4 and concentrated. The crude material was
purified by a 4 g silica gel column eluted with 30-50% EtOAc in
heptane to give methyl
2-(4-(4-(4-chloro-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-yl)-
-3-fluorophenyl)cyclohexyl)acetate a colorless oil (20 mg).
m/z=448.4(M+1).
Preparation 4A
Methyl
2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)a-
cetate
##STR00033##
[0161] Methyl
2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)acetate
was prepared as follows.
[0162] NaH (9.2 g, 384 mmol, 1.5 eq) was suspended in THF (850 ml).
This was cooled to .degree. C. and trimethylphosphonoacetate (40
ml, 277 mmol, 1.1 eq) was added dropwise keeping T<10.degree. C.
Mixture was then recooled to 0.degree. C. and stirred for 15 min.
Then compound 1 (40 g, 256 mmol, 1.0 eq) in THF (300 ml) was added
dropwise. The reaction mixture was stirred over night allowing it
to warm to room temperature. The mixture was poured into water (500
ml) and EtOAc (300 ml) were added, the layers were separated. The
aqueous layer was extracted with EtOAc (400 ml). The combined
organic layers were dried (Na.sub.2SO.sub.4) and evaporated. Yield:
51.6 g of methyl 2-(4-(1,3-dioxalane)cyclohexylidene)acetate;
.sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 1.78 (q, 4H), 2.37 (t,
2H), 2.99 (t, 2H), 3.68 (s, 3H), 3.99 (s, 4H), 5.65 (s, 1H)
[0163] Methyl 2-(4-(1,3-dioxalane)cyclohexylidene)acetate (51.6 g)
and 10% Pd/C (50% wet, (5 table-spoons)) was added to EtOAc (1 L).
The reaction mixture was degassed and a H.sub.2-balloon was
mounted. This was stirred over night at room temperature. The
reaction mixture was filtered over celite and rinsed with EtOAc.
The filtrate was evaporated. Yield: 51.1 g of methyl
2-(4-(1,3-dioxalane)cyclohexyl)acetate. .sup.1H NMR (CDCl.sub.3,
300 MHz) .delta. 1.35 (m, 2H), 1.57 (m, 2H), 1.76 (m, 4H), 1.82 (m,
1H), 2.22 (d, 2H), 3.65 (s, 3H), 3.95 (s, 4H)
[0164] Methyl 2-(4-(1,3-dioxalane)cyclohexyl)acetate (51.1 g, 238
mmol, 1.0 eq) was dissolved in THF (2 L). This was cooled to
0.degree. C. and 2 N HCl (1.4 L, 2.8 mol, 12.0 eq) was added
dropwise in 2.5 hours keeping T=0.degree. C. The reaction mixture
was stirred over night allowing it to warm to room temperature.
Then water (1 L) and EtOAc (1 L) were added and the layers were
separated. The aqueous layer was extracted with EtOAc (1 L). The
combined organic layers were washed with brine (1 L) and dried
(Na.sub.2SO.sub.4). The mixture was evaporated and purified by
column chromatography (silica: EtOAc/heptane=1/4). Yield: 21.9 g,
54% (clear oil) of methyl 2-(4-oxocyclohexyl)acetate. .sup.1H NMR
(CDCl.sub.3, 300 MHz) .delta. 1.50 (m, 2H), 2.16 (m, 2H), 2.36 (d,
2H), 2.38 (m, 1H), 2.39 (m, 4H), 3.71 (s, 3H)
[0165] Methyl 2-(4-oxocyclohexyl)acetate (8.13 g, 47.9 mmol, 1.0
eq) and 2,-di-tert.butyl-4-methylpyridine (11.3 g, 55 mmol, 1.15
eq) were dissolved in dry DCM (300 ml). To the reaction mixture
Tf.sub.2O (8.35 ml, 50 mmol, 1.05 eq) was added dropwise and
stirred for 2 h. The reaction mixture turned from orange to green
while adding Tf.sub.2O and after 30 min a suspension was formed.
The suspension was filtered and the filtrate was evaporated. Crude
mixture was dissolved in DCM (200 ml) and filtered. The filtrate
was evaporated. Yield: 14.3 g, 99% (green semi-solid) of methyl
2-(4-(trifluoromethyl-sulfonyloxy)cyclohex-3-enyl)acetate. .sup.1H
NMR (CDCl.sub.3, 300 MHz) .delta. 1.92 (m, 2H), 2.18 (m, 2H), 2.31
(d, 2H), 2.36 (m, 2H), 2.42 (m, 1H), 3.65 (s, 3H), 5.74 (t, 1H)
[0166] Methyl
2-(4-(trifluoromethyl-sulfonyloxy)cyclohex-3-enyl)acetate (21.9 g,
72.4 mmol, 1.0 eq), bis(pinacolato)diboron (16.7 g, 65.8 mmol, 0.9
eq), KOAc (21.5 g, 219 mmol, 3.0 eq) and PdCl.sub.2(dppf) (1.6 g,
2.19 mmol, 0.03 eq) were dissolved in DMSO (550 ml). The reaction
mixture was bubbled through with N.sub.2-gas for 15 min and then
heated to 50.degree. C. and stirred for 2.5 h. To the reaction
mixture water (400 ml) and EtOAc (600 ml) were added and the layers
were separated. The aqueous layer was extracted with EtOAc (600
ml). The combined organic layers were washed with brine (500 ml)
and dried (Na.sub.2SO.sub.4). The mixture was evaporated and
purified by column chromatography (silica: EtOAc/heptane=5/95).
Yield: 9.15 g, 45% (brown-yellow oil) of methyl
2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)acetate-
. .sup.1H NMR (CDCl.sub.3, 300 MHz) .delta. 1.25 (s, 12H), 1.77 (m,
2H), 1.83 (m, 1H), 2.07 (m, 1H), 2.14 (m, 1H), 2.20 (m, 2H), 2.56
(d, 2H), 3.67 (s, 3H), 6.50 (t, 1H).
EXAMPLE 1
Methyl{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6-
-(5H)-yl)phenyl]cyclohexyl}acetate
##STR00034##
[0168] The compound methyl
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6-(5H)-
-yl)phenyl]cyclohexyl}acetate, shown above, was prepared as
follows.
[0169] A solution of methyl
{trans-4-[4-(4-chloro-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6-(5H-
)-yl)phenyl]cyclohexyl}acetate, from Preparation 1, (5.29 g, 12.3
mmol) in 0.5M ammonia in p-dioxane (120 mL) was stirred at room
temperature for 24 hours. The reaction mixture was concentrated in
vacuo, diluted into EtOAc (1 L), washed with water, saturated
aqueous brine, dried over sodium sulfate and concentrated in vacuo
to afford the title compound as an off-white solid, 5.04 g. .sup.1H
NMR (400 MHz, CDCl.sub.3): .delta. 8.22 (s, 1H), 8.16 (br s, 1H),
7.23 (d, 2H), 7.16 (d, 2H), 5.75 (br s, 1H), 4.63 (m, 2H), 3.98 (m,
2H), 3.64 (s, 3H), 2.44 (m, 1H), 2.21 (m, 2H), 1.81 (m, 5H), 1.42
(m, 2H), 1.10 (m, 2H). m/z=411.3 (M+1). IC.sub.50 34.5 nM (range
30-40 nM).
EXAMPLE 2
{Trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-y-
l)phenyl]cyclohexyl}acetic acid
##STR00035##
[0171] The compound
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6-(5H)-
-yl)phenyl]cyclohexyl}acetate, shown above, was prepared as
follows.
[0172] A stirred solution of methyl
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6-(5H)-
-yl)phenyl]cyclohexyl}acetate (5.05 g, 12.3 mmol), from Example 1,
and 1N aqueous lithium hydroxide (36.9 mL) in p-dioxane (96 mL) and
water (27 mL) was stirred at 50.degree. C. for one hour. After
cooling, the reaction solution was adjusted to pH.about.3.5 with 6N
aqueous hydrochloric acid and the mixture was concentrated to near
dryness. This residue was slurried in water (40 mL) for 1 hour,
filtered, the solids washed with water (2.times.20 mL), ether
(3.times.30 mL) and dried in vacuo to afford the title compound as
an off-white solid, 4.58 g. .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. 8.12 (s, 1H), 7.58 (br s, 2H) 7.21 (s, 4H), 4.56 (m, 2H),
3.92 (m, 2H), 2.42 (m, 1H), 2.08 (m, 2H), 1.75 (m, 5H), 1.42 (q,
2H), 1.05 (q, 2H). m/z=397.3 (M+1). IC.sub.50 19.1 nM (range
5.2-63.6 nM).
EXAMPLE 3
(Trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]ox-
azepin-6(5H)-yl]phenyl}cyclohexyl)acetic acid
##STR00036##
[0174] The compound
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H )-yl]phenyl}cyclohexyl)acetic acid was prepared as
follows.
[0175] A mixture of methyl
(trans-4-{4-[(8R)-4-chloro-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-
oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetate, from Preparation 2,
(1.50 g, 3.38 mmol) in 0.5M ammonia in dioxane (20 mL) was stirred
at 50.degree. C., in a tightly capped flask, for 6 hours. The
reaction mixture was concentrated to give a white solid, which was
carried on to the next step without further purification. LiOH (247
mg, 9.89 mmol) was added to a solution of the white solid in
THF/MeOH/water (30 mL, 3:2:1) and then the resulting solution was
stirred at 23.degree. C. for 18 hours. 1M HCl solution was added to
reaction solution to adjust pH to about 3. 20% i-propanol in DCM
(130 mL) was added to extract reaction mixture. The organic layer
was separated and dried over MgSO.sub.4 and concentrated to give a
solid. Purification was done by chromatography (80 g, silica gel
column) with methanol/DCM from 2-6% to give a white solid 1210 mg
(89%) as the title compound. m/z=411.1(M+1). 1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 1.11-1.25 (m, 2H) 1.36 (d, J=6.64 Hz,
3H) 1.53 (q, J=12.88 Hz, 2H) 1.75-1.96 (m, 5H) 2.21 (d, J=7.03 Hz,
2H) 2.46-2.58 (m, 1H) 3.80-3.96 (m, 2H) 4.92-5.03 (m, 1H) 7.25 (d,
2H) 7.31 (d, 2H) 8.17 (s, 1H). IC.sub.50 19.3 nM (range 7.0-30.4
nM).
EXAMPLE 4
Methyl
(trans-4-{4-[(7S)-4-amino-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f]-
[1,4]oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetate
##STR00037##
[0177] The compound methyl
(trans-4-{4-[(7S)-4-amino-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl}cyclohexyl)acetate was prepared as
follows.
[0178] Methyl
(trans-4-{4-[(7S)-4-chloro-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-
oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetate (0.22 g, 0.496 mmol),
from Preparation 2, was diluted in 24.8 ml of 0.5M ammonia in
dioxane and heated to 50.degree. C., in a tightly capped vial for
four hours. After completion, reaction was concentrated to recover
0.26 g of methyl
(trans-4-{4-[(7S)-4-amino-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl cyclohexyl)acetate. 1H NMR (400 MHz,
chloroform-d) .delta. ppm 1.04-1.26 (m, 2 H) 1.33 (d, J=7.05 Hz,
3H) 1.40-1.60 (m, 2H) 1.74-2.04 (m, 5H) 2.26 (d, J=6.64 Hz, 2H)
2.43-2.64 (m, 1H) 3.68 (s, 3H) 3.95-4.16 (m, 1H) 4.46-4.73 (m, 2H)
7.10 (d, J=8.29 Hz, 2H) 7.29 (d, J=8.29 Hz, 2H) 8.22 (s, 1H)
m/z=425.1 (M+1).
EXAMPLE 5
(Trans-4-{4-[(7S)-4-amino-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]ox-
azepin-6(5H)-yl]phenyl cyclohexyl)acetic acid
##STR00038##
[0180] The compound
(trans-4-{4-[(7S)-4-amino-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl cyclohexyl)acetic acid was prepared as
follows.
[0181] Lithium hydroxide (11.3 mg, 0.472 mmol) was added to a
solution of methyl
(trans-4-{4-[(7S)-4-amino-7-methyl-5-oxo-7,8-dihydropyrimido[5,4-f-
][1,4]oxazepin-6(5H)-yl]phenyl cyclohexyl)acetate (50 mg, 0.12
mmol), from Example 4, in 2.76 ml of THF/methanol/water (3:2:1) and
the resulting solution was stirred at room temperature. After
completion, reaction was acidified with 1N NaOH until acidic pH,
and concentrated reaction mixture. EtOAc and water were added, and
repulped product for two hours. Filtered and dried to recover 2.1
mg of product. 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 0.99-1.19
(m, 2H) 1.08 (d, 3H) 1.33-1.53 (m, 2H) 1.57-1.88 (m, 5H) 2.06 (d,
J=6.65 Hz, 2H) 2.30-2.61 (m, 1H) 3.93-4.19 (m, 1H) 4.38-4.69 (m,
2H) 7.13 (d, J=8.31 Hz, 2H) 7.26 (d, J=8.31 Hz, 2H) 8.07 (s, 1H)
m/z=411.2 (M+1). IC.sub.50 33.9 nM.
EXAMPLE 6
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-yl)-3-
-fluorophenyl)cyclohexyl)acetic acid
##STR00039##
[0183] The title compound was prepared as follows. A mixture of
methyl
2-(4-(4-(4-chloro-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-yl)-
-3-fluorophenyl)cyclohexyl)acetate (20 mg, 0.045 mmol), from
Preparation 4, in 0.5M ammonia in dioxane was stirred at 50.degree.
C., in a tightly capped flask, for 3 hours. The reaction mixture
was concentrated to give a white solid, which was carried on to the
next step without further purification. LiOH (4.5 mg, 0.18 mmol)
was added to a solution of the white solid in THF/methanol/water
(2.4 mL, 3:2:1) and the resulting solution was stirred at
23.degree. C. for 16 hours. 1M HCl solution was added to reaction
solution to adjust the pH about 3. 20% i-propanol in DCM (30 mL)
was added to extract reaction mixture. The organic layer was
separated and dried over MgSO.sub.4 and concentrated to give a
solid. Purification was done by chromatography (4 g, silica gel
column) with methanol/DCM from 2-6% to give a white solid 12 mg as
the title compound. .sup.1H NMR (400 MHz, CD3OD): .delta. 8.18 (s,
1H), 7.28 (m, 1H), 7.16 (m,2H), 4.66 (m, 2H), 3.98 (m, 2H),
2.64-2.5 (m, 1H), 2.4, 2.2 (d, 2H), 2.22 (m, 1H), 1.90 (m, 2H), 1.7
(m, 4H), 1.54 (m, 2H), 1.2 (m, 2H). m/z=415.4(M+1). IC.sub.50 55.5
nM (range 29.1-89.6 nM).
[0184] The compounds of the Examples 7-16, having the following
structure, were prepared using the procedures described in Examples
1-4.
##STR00040##
EXAMPLE 7
[0185] (R.sup.1.dbd.H, R.sup.2.dbd.CH.sub.3, R.sup.3.dbd.CH.sub.3,
R.sup.4.dbd.H, R.sup.9.dbd.CH.sub.2CH.sub.3)-ethyl
{trans-4-[4-(4-amino-8,8-dimethyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]ox-
azepin-6(5H)-yl)phenyl]cyclohexyl}acetate m/z=453.3 (M+1) m/z=453.3
(M+1); IC.sub.50 183 nM
EXAMPLE 8
[0186] (R.sup.1.dbd.H, R.sup.2.dbd.CH.sub.3, R.sup.3.dbd.CH.sub.3,
R.sup.4.dbd.H,
R.sup.9.dbd.H)-{trans-4-[4-(4-amino-8,8-dimethyl-5-oxo-7,8-dihydropyrimid-
o[5,4-f][1,4]oxazepin-6(5H)-yl)phenyl]cyclohexyl}acetic acid
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.18 (s, 1H), 7.59 (br
s, 1H) 7.20 (m, 4H), 3.79 (m, 2H), 2.42 (m, 1H), 2.08 (m, 2H), 1.78
(m, 4H), 1.67 (m, 1H), 1.43 (m, 2H), 1.24 (s, 6H), 1.05 (m, 2H).
m/z=425.3 (M+1). IC.sub.50 146 nM (range 125.0-170.0 nM)
EXAMPLE 9
[0187] (R.sup.1.dbd.CH.sub.3, R.sup.2.dbd.H, R.sup.3.dbd.H,
R.sup.4.dbd.H, R.sup.9.dbd.CH.sub.3)-methyl
{trans-4-[4-(4-amino-2-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}acetate. 1H NMR (400 MHz,
CHLOROFORM-d) ppm .delta. 1.04-1.32 (m, 2H) 1.39-1.56 (m, 2H)
1.77-1.98 (m, 5H) 2.15-2.32 (m, 2H) 2.38 (s, 0H) 2.41-2.60 (m, 4H)
3.68 (s, 3H) 3.91-4.03 (m, 2H) 4.56-4.73 (m, 2H) 5.60 (br. s., 1H)
7.12-7.22 (m, 2H) 7.22-7.36 (m, 2H) 8.23 (br. s., 1H); m/z=425.3
(M+1). IC.sub.50 239 nM
EXAMPLE 10
[0188] (R.sup.1.dbd.CH.sub.3, R.sup.2.dbd.H, R.dbd.H,
R.sup.4.dbd.H,
R.sup.9.dbd.H)-{trans-4-[4-(4-amino-2-methyl-5-oxo-7,8-dihydropyrimido[5,-
4-f][1,4]oxazepin-6(5H)--I)phenyl]cyclohexyl}acetic acid .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 6.98 (m, 2H) 6.85 (m, 2H), 4.43
(m, 2H), 3.78 (m, 2H), 2.20 (m, 1H), 2.08 (s, 3H), 1.90 (m, 2H),
1.59 (m, 5H), 1.20 (m, 2H), 0.83 (m, 2H). m/z=411.4 (M+1).
IC.sub.50 59.4 nM (range 49.9-78.2 nM)
EXAMPLE 11
[0189] (R.sup.1.dbd.H, R.sup.2.dbd.CH.sub.3, R.sup.3.dbd.H,
R.sup.4.dbd.H,
R.sup.9.dbd.H)-{4-[4-(4-Amino-8-methyl-5-oxo-7,8-dihydro-5H-9-oxa-1,3,6-t-
riaza-benzocyclohepten-6-yl)-phenyl]-cyclohexyl}-acetic acid 1H NMR
(400 MHz, METHANOL-d.sub.4) .delta. ppm 1.11-1.27 (m, 2H) 1.37 (d,
J=4.98 Hz, 3H) 1.46-1.62 (m, 2H) 1.76-2.00 (m, 5H) 2.21 (d, J=5.39
Hz, 2H) 2.45-2.61 (m, 1H) 3.80-3.98 (m, 2H) 4.93-5.05 (m, 1H)
7.20-7.28 (m, 2H) 7.29-7.36 (m, 2H) 8.17 (s, 1H) m/z=411.4, 409.5
(M+1). IC.sub.50 125 nM (range 52.8-492.0 nM)
EXAMPLE 12
[0190] (R.sup.1.dbd.H, R.sup.2=CH.sub.3, R.sup.3.dbd.H,
R.sup.4.dbd.H,
R.sup.9.dbd.H)-{4-[4-((S)-4-Amino-8-methyl-5-oxo-7,8-dihydro-5H-9-oxa-1,3-
,6-triaza-benzocyclohepten-6-yl)-phenyl]-cyclohexyl}-acetic acid
.sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm 1.15-1.25 (m,
2H) 1.37 (d, J=6.64 Hz, 3H) 1.53 (q, J=12.75 Hz, 2H) 1.77-1.96 (m,
5H) 2.21 (d, J=7.03 Hz, 2H) 2.45-2.59 (m, 1H) 3.85-3.92 (m, 2H)
4.94-5.04 (m, 1H) 7.22-7.28 (m, 2H) 7.28-7.35 (m, 2H) 8.17 (s, 1H);
m/z=411.4, 409.5 (M+1). IC.sub.50 625 nM
EXAMPLE 13
[0191] (R.sup.1.dbd.H, R.sup.2.dbd.CH.sub.2CH.sub.3, R.sup.3.dbd.H,
R.sup.4.dbd.H,
R.sup.9.dbd.H)-{4-[4-(4-Amino-8-ethyl-5-oxo-7,8-dihydro-5H-9-oxa-1,3,6-tr-
iaza-benzocyclohepten-6-yl)-phenyl]-cyclohexyl}-acetic acid
m/z=425.2 (M+1) IC.sub.50 1330 nM (range 1070-1660 nM)
EXAMPLE 14
[0192] (R.sup.1.dbd.H, R.sup.2.dbd.H, R.sup.3.dbd.H,
R.sup.4=CH.sub.3,
R.sup.9.dbd.H)-(trans-4-{4-[(7R)-4-amino-7-methyl-5-oxo-7,8-dihydropyrimi-
do[5,4-f][1,4]-oxazepin-6(5H)-yl]phenyl cyclohexyl)acetic acid
.sup.1H NMR (400 MHz, DMSO-d6) .delta. ppm 1.00-1.16 (m, 2H) 1.08
(d, 3H) 1.34-1.52 (m, 2H) 1.67-1.86 (m,5H) 2.11 (d, J=7.06 Hz, 2H)
2.36-2.45 (m, 1H) 4.00-4.11 (m, 1H) 4.44-4.54 (m, 1H) 4.61 (d,
J=11.22 Hz, 1H) 7.14 (d, J=8.31 Hz, 2H) 7.26 (d, J=8.31 Hz, 2H)
8.07 (s, 1H), m/z=411.2 (M+1). IC.sub.50 24.7 nM (range 20.2-36.7
nM)
EXAMPLE 15
[0193] (R.sup.1.dbd.H, R.sup.2.dbd.H, R.sup.3.dbd.H,
R.sup.4.dbd.CH.sub.3,
R.sup.9.dbd.H)-{4-[4-(4-Amino-7-methyl-5-oxo-7,8-dihydro-5H-9-oxa-1,3,6-t-
riaza-benzocyclohepten-6-yl)-phenyl]-cyclohexyl}-acetic acid 1H NMR
(400 MHz, METHANOL-d4) .delta. ppm 1.08-1.34 (m, 2H) 1.26 (d, 3H)
1.45-1.63 (m, 2H) 1.76-2.01 (m, 5H) 2.22 (d, J=7.06 Hz, 2H) 2.54
(t, J=7.69 Hz, 1H) 4.16-4.37 (m, 1H) 4.65-4.87 (m, 2H) 7.17(d,
J=8.31 Hz, 2H) 7.33 (d, J=8.31 Hz, 2H) 8.24 (s, 1H) m/z=411.2
(M+1). IC.sub.50 27.3 nM (range 13.6-77.9 nM)
EXAMPLE 16
[0194] shown below,
{cis-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-yl-
)phenyl]cyclohexyl}acetic acid
##STR00041##
IC.sub.50 18.1 nM (range 16.0-20.5 nM)
[0195] The compounds of Examples 17-25, having one of the following
structures, using starting materials prepared analogous to the
method for preparing the compound of Preparation 4, were prepared
my the method of Scheme 1 by the methods of Examples 3 and 6.
TABLE-US-00001 ##STR00042## DGAT-1 DGAT-1 IC.sub.50 Range Ex.
R.sup.3 R.sup.5d R.sup.5a IC.sub.50 (nM) (nM) 17 CH.sub.3 H F
<6.21 <3-15.5 18 H H Cl 5.82 4.4-6.9 19 H H F <8.26
<3-15.5 20 H H CH.sub.3 13 8.6-17.2 21 CH.sub.3 H CH.sub.3 14.8
6.1-37.4 22 CH.sub.3 F H 30 27.9-34.5 23 H CH.sub.3 H 1410
643-2270
EXAMPLE 17
[0196]
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-
oxazepin-6(5H)-yl)-2-fluorophenyl)cyclohexyl)acetic acid (400 MHz,
CD3OD): .delta. 8.18 (s, 1H), 7.38 (m, 1H), 7.10 (m, 2H), 4.96 (m,
1H), 3.90 (m, 2H), 2.9 (m, 1H), 2.8, 2.3 (m, 1H), 2.4, 2.2 (d, 2H),
1.90 (m, 2H), 1.7 (m, 2H), 1.54 (m, 2H), 1.4 (d, 3H), 1.2 (m,
2H)
EXAMPLE 18
[0197]
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H-
)-yl)-2-chlorophenyl)cyclohexyl)acetic acid (400 MHz, CD3OD):
.delta. 8.18 (s, 1H), 7.42 (m, 2H), 7.14 (m, 1H), 4.66 (m, 2H),
3.98 (m, 2H), 3.0 (m, 1H), 2.3 (m, 1H), 2.44, 2.2 (d, 2H),
1.95-1.42 (m, 6H), 1.2 (m, 1H)
EXAMPLE 19
[0198]
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H-
)-yl)-2-fluorophenyl)cyclohexyl)acetic acid (400 MHz, CD3OD):
.delta. 8.18 (s, 1H), 7.38 (m, 1H), 7.1 (m, 2H), 4.68 (m, 2H), 4.02
(m, 2H), 2.9 (m, 1H), 2.24 (m, 1H), 2.4, 2.2 (d, 2H), 1.90 (m, 2H),
1.7 (m, 2H), 1.54 (m, 2H), 1.2 (m, 2H)
EXAMPLE 20
[0199]
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H-
)-yl)-2-methylphenyl)cyclohexyl)acetic acid (400 MHz, CD3OD):
.delta. 8.18 (s, 1H), 7.32 (m, 1H), 7.08 (m, 2H), 4.68 (m, 2H),
3.98 (m, 2H), 2.78 (m, 1H), 2.38 (s, 3H), 2.32 (m, 1H), 2.4, 2.2
(d, 2H), 1.90-1.5 (m, 7H), 1.2 (m, 7H)
EXAMPLE 21
[0200]
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-
oxazepin-6(5H)-yl)-2-methylphenyl)cyclohexyl)acetic acid (400 MHz,
CD3OD): .delta. 8.18 (s, 1H), 7.36 (m, 1H), 7.10 (m, 2H), 4.96 (m,
1H), 3.92 (m, 2H), 2.78 (m,1H), 2.34 (s, 3H), 2.3 (m, 1H), 2.5, 2.2
(d, 2H), 1.90 (m, 1H), 1.7 (m, 3H), 1.54 (m, 3H), 1.38 (d, 3H), 1.2
(m, 1H)
EXAMPLE 22
[0201]
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-
oxazepin-6(5H)-yl)-3-fluorophenyl)cyclohexyl)acetic acid (400 MHz,
CD3OD): .delta. 8.18 (s, 1H), 7.28 (m, 1H), 7.08 (m, 2H), 4.96 (m,
1H), 3.92 (m, 2H), 2.6 (m, 1H), 2.4, 2.2 (d, 2H), 2.3 (m, 1H), 1.90
(m, 2H), 1.7 (m, 4H), 1.5 (m, 1H), 1.4 (d, 3H), 1.2 (m, 1H)
EXAMPLE 23
[0202]
2-(4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]-oxazepin-6(5-
H)-yl)-3-methylphenyl)cyclohexyl)acetic acid-1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 1.12-1.25 (m, 1H) 1.45-1.59 (m, 1H)
1.62-1.76 (m, 3H) 1.88 (t, J=15.61 Hz, 3H) 2.22 (d, J=3.51 Hz, 3H)
2.26 (d, J=3.90 Hz, 1H) 2.43 (d, J=7.81 Hz, 2H) 2.46-2.55 (m, 1H)
3.78-4.05 (m, 2H) 4.60-4.78 (m, 2H) 7.07-7.26 (m, 3H) 8.16 (s, 1H);
LCMS was 411.4(t=2.0 min).
EXAMPLES 24-25
TABLE-US-00002 ##STR00043## [0203] Ex. ##STR00044## R.sup.3
R.sup.5d R.sup.5a DGAT-1IC.sub.50 (nM) DGAT-1IC.sub.50Range(nM) 24
Cis CH.sub.3 H F 4.6 1.7-13.2 25 Trans CH.sub.3 H F 12.3
10.0-17.0
EXAMPLE 24
[0204]
(cis4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,-
4]oxazepin-6(5H)-yl]-2-fluorophenyl}cyclohexyl)acetic acid (400
MHz, CD3OD): .delta. 8.18 (s, 1H), 7.38 (m, 1H), 7.10 (m, 2H), 4.96
(m, 1H), 3.90 (m, 2H), 2.9 (m, 1H), 2.4 (d, 2H), 2.3 (m, 1H), 1.90
(m, 1H), 1.7-1.5 (m, 6H), 1.4 (d, 3H), 1.2 (m, 1H)
EXAMPLE 25
[0205]
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f]-
[1,4]oxazepin-6(5H)-yl]-2-fluorophenyl}cyclohexyl)acetic acid (400
MHz, CD3OD): .delta. 8.18 (s, 1H), 7.38 (m, 1H), 7.10 (m, 2H), 4.96
(m, 1H), 3.88 (m, 2H), 2.8 (m, 1H), 2.2 (d, 2H), 1.90 (m, 5H), 1.58
(q, 2H), 1.4 (d, 3H), 1.2 (m, 2H).
EXAMPLE 26
2-{Trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-
-yl)phenyl]cyclohexyl}-N-(methylsulfonyl)acetamide
##STR00045##
[0207] The compound
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H-
)-yl)phenyl]cyclohexyl}-N-(methylsulfonyl)acetamide was prepared as
follows.
[0208] To a stirred solution of
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}acetic acid (50 mg, 0.13 mmol), from Example
2, N-methylsulfonamide (30 mg, 0.32 mmol) were added HOBT (29 mg,
0.19 mmol), EDCl (30 mg, 0.32 mmol) and TEA (19.1 mg, 0.19 mmol).
After 18 hours, the reaction mixture was diluted into EtOAc, washed
with water, dried over sodium sulfate and concentrated in vacuo.
Reverse-phase chromatography afforded the desired product as a
white solid, 5.3 mg. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
8.17 (s, 1H), 7.64 (br s, 2H) 7.20 (s, 4H), 4.58 (m, 2H), 3.92 (m,
2H), 3.20 (s, 3H), 2.42 (m, 1H), 2.17 (m, 2H), 1.73 (m, 5H), 1.40
(m, 2H), 1.05 m, 2H). m/z=474.4 (M+1). IC.sub.50 44.5 nM (range
34.2-75.0 nM).
EXAMPLE 27
(Trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]ox-
azepin-6(5H)-yl]phenyl}cyclohexyl)acetamide
##STR00046##
[0210] The compound
2-((1S,4s)-4-(4-((R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,-
4]oxazepin-6(5H)-yl)phenyl)cyclohexyl)acetamide was prepared as
follows.
[0211]
(Trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f]-
-[1,4]oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetic acid (1100 mg,
2.68 mmol), from Example 3, was added into DCM (20 mL) and DMF
(0.02 mL). Oxalyl chloride (0.47 mL, 5.36 mmol) was then added
dropwise at 0.degree. C., and the resulting mixture was warmed to
room temperature and stirred for 2 hours. The mixture was
concentrated to afforded a pale yellow solid (acyl chloride) and
dried on a vacuum pump for 1 hour. The acyl chloride was then
dissolved in 30 mL of 0.5N NH.sub.3 in dioxane and the reaction was
stirred for 90 minutes. 20% i-propanol in DCM (130 mL) and water
(100 mL) were added to reaction mixture. The organic layer was
separated and dried over MgSO.sub.4 and concentrated to give a
solid. Purification was done by chromatography (80 g, silica gel
column) with methanol/DCM from 2-10% to give a white solid 950 mg
(87%) as the title compound. m/z=410.1 (M+1). 1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 1.10-1.24 (m, 2H) 1.36 (d, J=6.24 Hz,
3H) 1.45-1.60 (m, 2H) 1.73-1.95 (m, 5H) 2.12 (d, J=7.03 Hz, 2H)
2.52 (t, J=12.29 Hz, 1H) 3.80-3.96 (m, 2H) 4.92-5.03 (m, 1H)
7.22-7.27 (m, 2H) 7.27-7.34 (m, 2H) 8.17 (s, 1H). IC.sub.50 52.2 nM
(range 11.4-68.1 nM).
[0212] The compounds of the following Examples 28-46 were prepared
utilizing the method of Examples 26-27 wherein an amide is formed
by reacting a carboxylic acid with an amine. The compounds of
Examples 33-44 were separated using reverse phase column
chromatography utilizing YMC ODS-AQ (2.0.times.50 mm 5 .mu.m),
which is a reversed-phase packing material with both a hydrophobic
high carbon loading and a relatively hydrophilic surface (YMC Co.,
Ltd., Tokyo, Japan) and 0.05% TFA in water as an eluent.
EXAMPLE 28
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepi-
n-6(5H)-yl)-2-fluorophenyl)cyclohexyl)acetamide
##STR00047##
[0214] The titled compound was prepared by the methods of Examples
6 and 27. 1H NMR (400 MHz, DMSO): .delta. 8.18 (s, 1H), 7.6 (br s,
1H), 7.34 (m, 3H), 7.20 (m,2H), 6.72 (br s, 1H), 4.9 (m, 1H), 3.82
(m, 2H), 2.87 (m,1H), 2.2 (m, 3H), 1.7-1.44 (m, 8H), 1.2 (d, 3H).
IC.sub.50 11.3 nM (range 7.3-19.9 nM).
TABLE-US-00003 ##STR00048## IC.sub.50 DGAT-1 DGAT-1 Range Ex.
R.sup.10 IC.sub.50 (nM) Ex. R.sup.10 IC.sub.50 (nM) (nM) 29
S(O).sub.2CF.sub.3 57.9 30 H 31.7 11.4-68.1 31 Tetrazole 18.3 32
CH.sub.2CH.sub.3 93.7 68.6-135.0 33 Cyclohexyl 11.1 34 cyclopentyl
64.3 35 2-indanyl 24.9 36 t-butyl 164 65.1-394.0 37 ##STR00049##
81.5 38 ##STR00050## 54.1 39 ##STR00051## 56.8 40 ##STR00052## 51.5
41 ##STR00053## 65.3 42 ##STR00054## 69.9 43 ##STR00055## 55.8 44
CH.sub.3 85.5 41.1-72.1 45 ##STR00056## 43.5 46 i-propyl 76.9
58.1-104
EXAMPLE 29
[0215]
2-{Trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepi-
n-6(5H)-yl)phenyl]cyclohexyl}-N-[(trifluoromethyl)sulfonyl]acetamide
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.23 (s, 1H), 8.10 (br
s, 2H) 7.22 (m, 4H), 4.62 (m, 2H), 3.99 (m, 2H), 2.40 (m, 1H), 2.01
(m, 2H), 1.71 (m, 5H), 1.39 (m, 2H), 1.00 (m, 2H). m/z=528.4
(M+1)
EXAMPLE 30
[0216]
2-{Trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepi-
n-6(5H)-yl)phenyl]cyclohexyl}acetamide .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 8.17 (s, 1H), 7.59 (br s, 2H) 7.24 (s, 4H),
4.58 (m, 2H), 3.96 (m, 2H), 2.44 (m, 1H), 1.97 (m, 2H), 1.78 (m,
4H), 1.68 (m, 1H), 1.42 (m, 2H), 1.03 (m, 2H). m/z=396.2 (M+1)
EXAMPLE 31
[0217]
2-{Trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepi-
n-6(5H)-yl)phenyl]cyclohexyl}-N-1H-tetrazol-5-ylacetamide .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. 8.17 (s, 1H), 7.63 (br s, 2H)
7.22 (m, 4H), 4.57 (m, 2H), 3.90 (m, 2H), 2.40 (m, 1H), 2.33 (m,
2H), 1.78 (m, 5H), 1.40 (m, 2H), 1.10 (m, 2H). m/z=464.4 (M+1)
EXAMPLE 32
[0218]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepi-
n-6(5H)-yl)phenyl]cyclohexyl}-N-ethylacetamide .sup.1H NMR (400
MHz, DMSO-d6) .delta. ppm 8.11 (s, 1H) 7.74 (t, J=5.39 Hz, 1H) 7.55
(br. s., 2H) 7.18-7.27 (m, 4H) 4.49-4.57 (m, 2H) 3.86-3.95 (m, 2H)
2.94-3.06 (m, 2H) 2.36-2.48 (m, 1H) 1.92 (d, J=6.64 Hz, 1H) 1.73
(t, J=9.75 Hz, 6H) 1.32-1.48 (m, 2H) 0.88-1.09 (m, 5H)
EXAMPLE 33
[0219]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-cyclohexylacetamide Mass Spectrum
M/Z (M+1) 478; RT=2.843
EXAMPLE 34
[0220]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-cyclopentylacetamide Mass Spectrum
M/Z (M+1) 464; RT=2.6
EXAMPLE 35
[0221]
2-{trans-4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl)cyclohexyl}-N-2-indanyl-acetamide Mass Spectrum
M/Z (M+1) 512; RT=2.586
EXAMPLE 36
[0222]
2-{trans-4-(4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl)cyclohexyl}-N-tert-butylacetamide Mass Spectrum
M/Z (M+1) 452; RT=2.633
EXAMPLE 37
[0223]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-[(1S)-1,2,2-trimethylpropyl]acetamide
Mass Spectrum M/Z (M+1) 480; RT=2.597
EXAMPLE 38
[0224]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-(1,1-dimethylpropyl)acetamide Mass
Spectrum M/Z (M+1) 466; RT=2.763
EXAMPLE 39
[0225]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-(2,2-dimethylpropyl)acetamide Mass
Spectrum M/Z (M+1) 466; RT=2.498
EXAMPLE 40
[0226]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-[(2S)-2-methylbutyl]acetamide Mass
Spectrum M/Z (M+1) 466; RT=2.508
EXAMPLE 41
[0227]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-(1-ethylpropyl)acetamide Mass
Spectrum M/Z (M+1) 466; RT=2.463
EXAMPLE 42
[0228]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-(1,2-dimethylpropyl)acetamide Mass
Spectrum M/Z (M+1) 466; RT=2.991
EXAMPLE 43
[0229]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-[(1S)-1-methylbutyl]acetamide Mass
Spectrum M/Z (M+1) 466; RT=2.808
EXAMPLE 44
[0230]
2-{4-[4-(4-Amino-5-oxo-7,8-dihydro-5H-9-oxa-1,3,6-triaza-benzocyclo-
hepten-6-yl)-phenyl]-cyclohexyl}-N-methyl-acetamide 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 0.89-1.15 (m, 2H) 1.30-1.50 (m, 2H)
1.59-1.83 (m, 5H) 1.85-1.98 (m, 2H) 2.39-2.59 (m, 2H) 3.02-3.38 (m,
3H) 3.83-3.96 (m, 2H) 4.46-4.59 (m, 2H) 7.15-7.29 (m, 2H) 7.47-7.59
(m, 2H) 7.66 (br. s., 2H) 8.11 (s, 1H).
EXAMPLE 45
[0231]
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f]-[1,4]oxazep-
in-6(5H)-yl)phenyl]cyclohexyl}-N-(2-methylbutyl)acetamide Mass
Spectrum M/Z (M+1) 466; RT=2.822
EXAMPLE 46
{Trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-y-
l)phenyl]cyclohexyl}acetonitrile
##STR00057##
[0233] The compound
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}acetonitrile was prepared as follows.
[0234] To a stirred slurry of
2-{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H-
)-yl)phenyl]cyclohexyl}acetamide (45 mg, 0.11 mmol), from Example
30, in THF (1 mL) was added DMF (0.002 mL) and oxalyl chloride
(0.05 mL, 0.6 mmol). After two hours of stirring at room
temperature, the reaction was quenched with aqueous sodium
bicarbonate and extracted with EtOAc. The organic layer was washed
with water, dried over sodium sulfate and concentrated in vacuo to
afford the title compound as a light-yellow colored solid, 34 mg.
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.17 (s, 1H), 7.63 (br
s, 2H) 7.22 (m, 4H), 4.57 (m, 2H), 3.90 (m, 2H), 2.43 (m, 3H), 1.78
(m, 4H), 1.63 (m, 1H), 1.43 (m, 2H), 1.08 (m, 2H). m/z=378.3 (M+1).
IC.sub.50 64.3 nM (range 51.2-91.1 nM).
EXAMPLE 47
(Trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]ox-
azepin-6(5H)-yl]phenyl}cyclohexyl)acetonitrile
##STR00058##
[0236] The compound
(trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl]phenyl}cyclohexyl)acetonitrile was prepared as
follows.
[0237]
(Trans-4-{4-[(8R)-4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f]-
[1,4]oxazepin-6(5H)-yl]phenyl}cyclohexyl)acetic acid, from Example
27, (500 mg, 1.22 mmol) was dissolved in THF (12 mL) and DMF (0.01
mL, 0.12 mmol), and oxalyl chloride (0.5 mL, 6 mmol) was added
dropwise to it at room temperature and the resulting mixture was
stirred at room temperature for 2 hours. The reaction mixture was
quenched with a carefully added NaHCO.sub.3 solution and water, and
then was extracted with EtOAc (2.times.100 mL). The organic layers
were combined and washed with water, dried over MgSO.sub.4 and
concentrated to give some crude product. The material was
chromatographed on 40 g silica gel column with 1-5% methanol/DCM to
give 250 mg (53%) white solid as the title compound. m/z=392.2
(M+1). 1H NMR (400 MHz, chloroform-d) .delta. 1.19-1.37 (m, 2H)
1.42-1.58 (m, 5H) 1.69-1.84 (m, 1H) 1.90-2.05 (m, 4H) 2.31 (d,
J=6.65 Hz, 2H) 2.46-2.60 (m, 1H) 3.76-3.96 (m, 2H) 4.86-5.00 (m,
1H) 5.58 (br. s., 1H) 7.20 (d, 2H) 7.27 (d, 2H) 7.99 (br. s., 1H)
8.29 (s, 1H). IC.sub.50 38.4 nM (range 30.1-48.1 nM).
EXAMPLE 48
(R)-2-(4-(4-(4-amino-8-methyl-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepi-
n-6(5H)-yl)-2-fluorophenyl)cyclohexyl)acetonitrile
##STR00059##
[0239] The titled compound was prepared by the methods of Examples
6 and 47. 1H NMR (400 MHz, CD3OD): .delta. 8.18 (s, 1H), 7.38 (m,
1H), 7.10 (m,2H), 4.96 (m, 1H), 3.90 (m, 2H), 2.84 (m, 1H), 2.6,
2.4 (d, 2H), 1.96-1.56 (m, 4H), 1.4 (d, 3H), 1.28 (m, 1H), 0.88 (m,
4H). IC.sub.50 12.9 nM (range 4.5-22.2 nM).
EXAMPLE 49
4-[4-(4-Amino-5-oxo-7,8-dihydro-5H-9-oxa-1,3,6-triaza-benzocyclohepten-6-y-
l)-phenyl]-cyclohexanecarboxylic acid
##STR00060##
[0241] Prepared according the methods described in Example 1. 1H
NMR (400 MHz, DMSO-d6) .delta. ppm 1.37-1.55 (m, 4H) 1.81 (br. s.,
2H) 1.99 (br. s., 2H) 2.25 (br. s., 2H) 3.85-4.03 (m, 2H) 4.45-4.61
(m, 2H) 7.20-7.32 (m, 4H) 7.58 (s, 2H) 8.15 (s, 1H). m/z=381 (M-1).
IC.sub.50 7.8 nM (range 5.8-10.5 nM)
EXAMPLE 50
4-[4-(4-Amino-5-oxo-7,8-dihydro-5H-9-oxa-1,3,6-triaza-benzocyclohepten-6-y-
l)-phenyl]-cyclohexanecarboxylic acid methyl ester
##STR00061##
[0243] Prepared according the methods described in Example 1. 1H
NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.32-1.68 (m, 5H) 1.87-1.98
(m, 2H) 2.03-2.14 (m, 2H) 2.22-2.38 (m, 1H) 2.46-2.58 (m, 1H)
3.59-3.70 (m, 2H) 3.91-4.03 (m, 2H) 4.55-4.72 (m, 2H) 5.64 (br. s.,
1H) 7.10-7.30 (m, 4H) 8.14 (br. s., 1H) 8.25 (s, 1H). m/z=397.4
(M+1). IC.sub.50 47.3 nM (range 45.2-49.5 nM)
EXAMPLE 51
4-Amino-6-{4-[4-(1H-tetrazol-5-ylmethyl)-cyclohexyl]-phenyl}-7,8-dihydro-6-
H-9-oxa-1,3,6-triaza-benzocyclohepten-5-one
##STR00062##
[0245] To a cooled 0.degree. C., stirred solution of
trimethylaluminum (2M in toluene, 0.33 mL) in toluene (0.66 mL)
were added trimethylsilylazide (0.86 mL) and
{trans-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)--
yl)phenyl]cyclohexyl}acetonitrile (25 mg). This mixture was heated
at 80.degree. C. for 40 hours, cooled, concentrated in vacuo and
chromatographed (4 g silica gel column, 0-10% methanol:chloroform)
to afford the title compound as a white solid, 2.8 mg. 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 0.96-1.28 (m, 2H) 1.25-1.45 (m, 2H)
1.62-1.80 (m, 5H) 2.31-2.50 (m, 1H) 2.62-2.76 (m, 2H) 3.85-3.95 (m,
2H) 4.45-4.57 (m, 2H) 7.13-7.29 (m, 4H) 7.55 (br. s., 2H) 8.10 (s,
1H) m/z=421.3 (M+1). IC.sub.50 76.2 nM (range 47.5-163 nM).
[0246] The following compounds of the present invention, as
described in Examples 52-57, were prepared, or were derived from
compounds prepared, by the method of Scheme 2 using the compound of
Preparation 5.
Preparation 5
4-Amino-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-one
##STR00063##
[0248]
4-Amino-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(-
6H)-one was prepared as follows.
[0249] Step 1.
N-(2-{[Tert-butyl(dimethyl)silyl]oxy}ethyl)-4-iodoaniline, shown
below, was prepared as follows,
##STR00064##
[0250] To a stirred slurry of sodium hydride (464 mg 60% in oil,
11.6 mmol) in THF (5 mL) was added a solution of 4-iodoaniline
(1.27 grams, 5.80 mmol) in THF (1 mL) dropwise. After 15 minutes,
(2-bromoethoxy)(tert-butyl)dimethyl silane (1.39 g, 5.83 mmol) was
added and the resulting slurry was stirred for 40 hours. The
reaction was quenched with water, partitioned between EtOAc and
saturated aqueous ammonium chloride. The aqueous layer was washed
with EtOAc (2.times.) and the combined organic layers were
saturated aqueous brine, dried over magnesium sulfate and
concentrated in vacuo to afford an oil. Chromatography (40 grams
silica gel column, 0-20% EtOAc:heptane) afforded the title compound
as a gummy solid, 1.41 grams. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 7.41, (d, 2H), 6.40 (d, 2H), 4.08 (br s, 1H), 3.81 (m, 2H),
3.18 0.92 (s, 9H), 0.04 (d, 6H). m/z=378.2 (M+1).
[0251] Step 2.
4,6-Dichloro-N-(2-hydroxyethyl)-N-(4-iodophenyl)pyrimidine-5-carboxamide,
shown below, was prepared as follows,
##STR00065##
[0252] To a cooled (ice/water), stirred solution of
N-(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)-4-iodoaniline (1.34 g,
2.78 mmol) and diethylisopropylamine (0.7 mL, 4.0 mmol) in MTHF (5
mL) was added 4,6-dichloroprimidine-5-carbonyl chloride (560 mg,
2.65 mmol). After 5 minutes, the cooling bath was removed and the
slurry was stirred for 5 hours. Aqueous hydrochloric acid (1N, 5
mL) was added and the mixture was allowed to stand for 72 hours.
The layers were separated and the aqueous layer was washed with
MTHF (2.times.). The combined organic layers were washed with
saturated aqueous sodium bicarbonate, saturated aqueous brine,
dried over sodium sulfate and concentrated in vacuo to afford an
oil. Chromatography (40 g silica gel column, 20-100% EtOAc:heptane)
afforded the title compound, 542 mg. 1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.64 (s, 1H), 7.62 (d, 2H), 7.17 (d, 2H), 4.07 (t, 2H),
3.91 (q, 2H). m/z=438.0 (M+1).
[0253] Step 3.
4-Chloro-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-o-
ne, shown below, was prepared as follows.
##STR00066##
[0254] A stirred slurry of
4,6-dichloro-N-(2-hydroxyethyl)-N-(4-iodophenyl)pyrimidine-5-carboxamide
(543 mg, 1.24 mmol) and potassium carbonate (350 mg, 2.53 mmol) in
DMF was heated at 80.degree. C. for 2 hours. The reaction was
cooled, filtered, solids washed with small portion of DMF and the
combined filtrates were concentrated in vacuo. The residue was
chromatographed (40 g of silica gel, 0-70% EtOAc:heptane) to afford
4-chloro-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-o-
ne as a white solid, 427 mg. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.78 (s, 1H), 7.80 (d, 2H), 7.17 (d, 2H), 4.77 (m, 2H),
4.03 (m, 2H). m/z=402.0 (M+1).
[0255] Step 4. A solution of
4-chloro-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-o-
ne (279 mg, 0.70 mmol) in 0.5 N ammonia in p-dioxane was stirred in
a sealed tube for 18 hours. Concentration of the reaction mixture,
followed by chromatography (10 g silica gel, 0-5%
methanol:chloroform) afforded the title compound of Preparation 5
as a white solid, 122 mg. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.18 (s, 1H), 7.80 (d, 2H), 7.60 (br s, 2H), 7.19 (d, 2H),
4.60 (m, 2H), 3.96 (m, 2H). m/z=383.1 (M+1).
EXAMPLE 52
Methyl
(+)-{4-[4-(4-Amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(-
5H)-yl)phenyl]cyclohex-3-en-1-yl}acetate
##STR00067##
[0257] Methyl
(.+-.)-{4-[4-(4-Amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-
-yl)phenyl]cyclohex-3-en-1-yl}acetate, shown above, was prepared as
follows.
[0258] A stirred solution of
4-amino-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-on-
e (40 mg, 0.10 mmol),
methyl[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl]-
acetate (38 mg, 0.14 mmol), cesium carbonate (42 mg, 0.28 mmol) and
palladium tetrakis(triphenyl-phosphine) (6 mg, 0.005 mmol) in
1,2-dimethoxyethane (1.0 mL) was heated at 100.degree. C. for 18
hours. The reaction mixture was cooled, concentrated and
chromatographed to afford a white solid, 19 mg. IC.sub.50 84.3
nM.
EXAMPLE 53
(+)-{4-[4-(4-Amino-5-oxo-7,8-dihydropyrimido[5.4-f][1,4]oxazepin-6(5H-yl)p-
henyl]cyclohex-3-en-1-yl}acetic acid
##STR00068##
[0260] To form the named compound, shown above, the compound of
Example 52 was hydrolyzed utilizing the conditions described in
Example 2 to afford the title compound as light brown solid, 11 mg.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.07 (br. s., 1H),
8.16 (s, 1H), 7.61 (br. s., 2H), 7.44 (d, J=8.30 Hz, 2H), 7.31 (d,
2H), 6.13 (br. s., 1H), 4.58 (t, 2H), 3.97 (br. s., 2H), 2.42 (br.
s., 2H), 2.32 (d, 2H), 2.23 (d, 2H), 1.92-2.03 (m, 1H), 1.83-1.93
(m, 1H), 1.31-1.45 (m, 1H). m/z=395.3 (M+1). IC.sub.50 36.4 nM
(range 33.8-39.3 nM).
EXAMPLE 54
Methyl{4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-6(5H)-y-
l)phenoxy]phenyl}acetate
##STR00069##
[0262] A stirred slurry of
4-amino-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-on-
e (38 mg, 0.10 mmol), methyl 4-hydroxyphenylacetic acid (25 mg,
0.15 mmol), cesium carbonate (78 mg, 0.24 mmol), N,N-dimethyl
glycine (10.3 mg, 0.10 mmol) and copper (I) iodide (6 mg, 0.03
mmol) in p-dioxane (0.4 mL) were heated at 90.degree. C. for 18
hours. The reaction mixture was cooled, concentrated in vacuo and
chromatographed (0-5% methanol:chloroform) to afford the title
compound as a solid, 2.4 mg. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.31 (s, 1H), 7.25 (d, 2H), 7.20 (d, 2H), 7.07 (d, 2H),
7.02 (d, 2H), 4.69 (m, 2H), 4.00 (m, 2H), 3.68 (s, 3H). m/z=421.2
(M+1). IC.sub.50 254 nM.
EXAMPLE 55
.+-.-{(1R,3aS,4R,6aR)-4-[4-(4-Amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]o-
xazepin-6(5H)-yl)phenyl]octahydropentalen-1-yl}acetic acid
##STR00070##
[0264] The named compound, shown above, was prepared as
follows.
[0265] Step 1. Dimethyl
(.+-.)-{(1R,3aS,4R,6aS)-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,-
4]oxazepin-6(5H)-yl)phenyl]octahydropentalen-1-yl}malonate was
prepared as follows. A slurry of
4-amino-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-on-
e (207 mg, 0.54 mmol), 1,5-cyclo octadiene (119 mg, 1.1 mmol),
dimethylmalonate (347 mg, 2.6 mmol), tetra-N-butylammonium chloride
(165 mg, 0.59 mmol), sodium bicarbonate (284 mg, 3.4 mmol) and
dipalladium(0)tris(dibenzylideneacetone)-chloroform (26 mg, 0.02
mmol) in dimethylsulfoxide (6 mL) was stirred at 80.degree. C. for
65 hr. The reaction mixture was cooled, concentrated and
chromatographed to afford the title compound, 197 mg. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.25 (s, 1H), 7.29 (d, 2H), 7.18 (d,
2H), 4.65 (m, 2H), 3.75 (s, 3H), ), 3.72 (s, 3H), 3.38 (d, 1H),
2.80 (m, 1H), 2.43 (m, 2H), 1.97 (m, 1H), 1.75-1.32 (m, 7H).
m/z=495.3 (M+1).
[0266] Step 2.
(.+-.)-{(1R,3aS,4R,6aS)-4-[4-(4-Amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,-
4]oxazepin-6(5H)-yl)phenyl]octahydropentalen-1-yl}malonic acid was
prepared as follows. A solution of dimethyl
(.+-.)-{(1R,3aS,4R,6aS)-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,-
4]oxazepin-6(5H)-yl)phenyl]octahydropentalen-1-yl}malonate (190 mg,
0.38 mmol) and lithium hydroxide monohydrate (166 mg, 3.9 mmol) in
p-dioxane/water (8:3-11 mL) was stirred at 50.degree. C. for 3
hours. The reaction was concentrated to remove the p-dioxane, water
(2 mL) added and the mixture extracted with EtOAc (5 mL). The
aqueous layer was adjusted to pH.about.3 with 2N aqueous
hydrochloric acid, the solids filtered and dried in vacuo to afford
the title compound as a white solid, 179 mg. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.17 (s, 1H), 7.58 (br s, 2H), 7.13 (d, 2H),
7.10 (d, 2H), 4.57 (m, 2H), 3.95 (m, 2H), 3.08 (d, 1H), 2.64 (m,
1H), 2.39 (m, 2H), 2.24 (m, 1H), 1.87-1.10 (m, 7H). m/z=467.3
(M+1).
[0267] Step 3. The title compound was prepared as follows. A
solution of
(.+-.)-{(1R,3aS,4R,6aS)-4-[4-(4-amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,-
4]oxazepin-6(5H)-yl)phenyl]octahydropentalen-1-yl}malonic acid (87
mg, 0.19 mmol) in xylenes (3 mL) and dimethylsulfoxide (0.5 mL) was
stirred at 150.degree. C. for 2 hours. After cooling the reaction
mixture was diluted in EtOAc, washed with water and a precipitate
formed in the organic layer. This solid was filtered and dried
under vacuum to afford the title compound as a white solid, 24 mg.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.17 (s, 1H), 7.60 (br
s, 2H), 7.15 (d, 2H), 7.12 (d, 2H), 4.58 (m, 2H), 3.96 (m, 2H),
3.13 (s, 1H), 2.62-1.12 (m, 13H). m/z=423.3 (M+1). IC.sub.50<10
nM (range<3-27.2 nM).
EXAMPLE 56
(1R,5R,6S)-6-[4-(4-Amino-5-oxo-7,8-dihydro-5H-9-oxa-1,3,6-triaza-benzocycl-
ohepten-6-yl)-phenyl]-tricyclo[3.2.1.0*2.4*]octane-3-carboxylic
acid methyl ester
##STR00071##
[0269] The named was prepared as follows. A slurry of
4-amino-6-(4-iodophenyl)-7,8-dihydropyrimido[5,4-f][1,4]oxazepin-5(6H)-on-
e (340 mg, 0.89 mmol), tricycle[3.2.1.0*2,4*]oct-6-ene-3-carboxylic
acid ethyl ester (240 mg, 1.35 mmol), tetrabutylammonium iodide (67
mg, 0.18 mmol), piperidine (230 mg, 2.7 mmol), formic acid (170 mg,
0.14 mmol) and dichlorobis(acetonitrile) palladium(II) (23 mg, 0.09
mmol) in DMF (0.36 mL) was stirred at 125.degree. C. for 16 hours.
The reaction mixture was cooled, diluted into EtOAc, washed with
water, saturated aqueous brine, dried over magnesium sulfate and
concentrated in vacuo to afford an oil. Chromatography (20 g of
silica gel, 1-5% methanol:chloroform) afforded a solid, 127 mg.
IC.sub.50 78.3 nM.
EXAMPLE 57
(.+-.)-(1R,5R,6S)-6-[4-(4-Amino-5-oxo-7,8-dihydropyrimido[5,4-f][1,4]oxaze-
pin-6(5H)-yl)phenyl]tricyclo[3.2.1.0.about.2.4.about.]octane-3-carboxylic
acid
##STR00072##
[0271] The title compound was prepared from the compound of Example
56 by hydrolyzing the ester as described in Example 2 to afford an
off-white solid, 27 mg. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
8.19 (s, 1H), 7.75 (br s, 2H), 7.24 (m, 4H), 4.60 (m, 2H), 3.97 (m,
2H), 2.89 (s, 1H), 2.42 (m, 1H), 1.83 (m, 2H), 1.60 (m, 2H), 1.38
(AB pattern, 2H), 0.90 (s, 2H). m/z=407.4 (M+1). IC.sub.50 34.7 nM
(range 26.7-58.6 nM).
Biological Protocols
[0272] The utility of the compounds of formula (1), the
pharmaceutically acceptable salts of the compounds, (such as are
described herein) in animals, particularly mammals (e.g., humans)
may be demonstrated by the activity thereof in conventional assays
known to one of ordinary skill in the relevant art, including the
in vitro and in vivo assays discussed below. Such assays also
provide a means whereby the activities of the compounds of formula
(1) can be compared with the activities of other known
compounds.
In Vitro Assay for DGAT-1 Inhibition
[0273] Human full-length diacylglycerol:acylCoA acyltransferase 1
(DGAT-1) was expressed in Sf9 insect cells which are then lysed and
a crude membrane fraction (105,000.times.g pellet) was prepared.
The DGAT-1 gene is a human DGAT-1 gene described in J. Biol. Chem.
273:26765, 1998 and U.S. Pat. No. 6,100,077.
[0274] In vitro inhibition of DGAT-1 was measured using a
modification, further described below, of the assay methodology
described in U.S. Pat. No. 6,994,956 B2.
[0275] The cells were cultured as follows. Sf9 cells (20 L) were
infected with 4 mL of DGAT1 Baculovirus Infected Insect Cells
(BIIC) for 72 hours in a Wave Bioreactor System 20/50P (Wave
Biotec/GE Healthcare).
[0276] Crude DGAT-1 microsomes were prepared as follows. Cell
pellets were washed once with ice-cold Dulbecco's
phosphate-buffered saline. Cells were collected in tabletop
centrifuge (Beckman GS-6KR), 15 minutes, 2000.times.g, 4.degree. C.
Twenty (20) mL of ice-cold Microsome Buffer (MB) was added per 5 g
of cell pellet. The suspension was passed through a microfluidizer
3 times (18K psi). The lysate was transferred to centrifuge tubes
and centrifuged for 20 minutes at 5000.times.g (Beckman-Coulter,
Inc. Allegra.RTM. 64R High-Speed Refrigerated Benchtop Centrifuge,
F0650 rotor) at 4.degree. C. The supernatant was transferred to
ultracentrifuge tubes and centrifuged at 125,000.times.g for 1 hr
in a Beckman Ti-45 rotor, 4.degree. C. The supernatant fluid was
discarded. The pellet was resuspended in 70 mL of MB by sonication.
The microsome concentration was determined using Bio-Rad Protein DC
Protein Assay. The microsomes were filter sterilized with a 0.22
.mu.m filter. The samples were portioned, flash frozen and stored
at -80.degree. C.
[0277] The Microsome Buffer, used for microsome preparation, was
prepared by conventional means and contained 125 mM sucrose, 3 mM
imidazole, 0.2 .mu.g/mL aprotinin, 0.2 .mu.g/mL leupeptin and 5 mM
dithiothreitol (Cleland's reagent), DGAT-1 activity was measured in
384-well format in a total assay volume of 25 .mu.l that contained,
Hepes buffer (50 mM, pH 7.5), MgCl.sub.2 (10 mM), bovine serum
albumin (0.6 mg/ml), [.sup.14C]decanoylCoA (20 .mu.M, 58 Ci/mol)
and membranes (25 .mu.g/ml) into which 1,2 dioleoyl-sn-glycerol (75
.mu.M) in acetone has already been incorporated. Inhibitors in DMSO
were pre-incubated with membranes before initiating the DGAT-1
reaction by the addition of decanoylCoA. Two control DGAT-1
reactions were also incubated in parallel: 1) DMSO without
inhibitor to measure zero percent effect of inhibition and 2) and a
maximally inhibited DGAT-1 reaction ("blank") incubated with 1
.mu.M {trans-4-[4-(4-amino-2,7,7-trimethyl-7
H-pyrimido[4,5-b][1,4]oxazin-6-yl)phenyl]cyclohexyl}acetic acid
(WO2004/047755). The DMSO concentration was 2.5%. The inhibitors
were present at a range of eight concentrations to generate an
apparent IC.sub.50 for each compound. The eight inhibitor
concentration employed ranged from 10 .mu.M to 3 nM (from high to
low concentration). Specifically, the eight concentrations used
were 10 .mu.M, 3 .mu.M, 1 .mu.M, 300 nM, 100 nM, 30 nM, 10 nM and 3
nM.
[0278] The reactions were allowed to proceed for 1.5 h at room
temperature and then terminated by the addition of 10 .mu.l of HCl
(0.5 M). Reaction mixtures were neutralized by the addition of 15
.mu.l of tris(hydroxy-methyl)aminomethane (1M, pH 8.0) and then
mixed by trituration with 37.5 .mu.l of Microscint.TM.-E (Perkin
Elmer). Plates contents were allowed to partition for 15 to 30 min
before .sup.14C was measured in a scintillation spectrometer
(Wallac Microbeta Trilux 1450-030 liquid scintillation counter 12
detector in the top-count DPM mode). Percent inhibition of test
compounds was computed as 100-((DPM DMSO uninhibited-DPM test
compound)/(DPM DMSO uninhibited)).
[0279] The compounds of the present invention, described in
Examples 1-3 and 5-57, were tested for in vitro DGAT-1 inhibition,
and were found to generally exhibit DGAT-1 inhibition with
IC.sub.50 values of 1000 nM or less. Where this DGAT-1 inhibition
assay was performed on a compound more than once, an inhibition
range is also provided for that compound. Preferably, the compounds
of the present invention exhibit DGAT-1 inhibition with IC.sub.50
values of 100 nM or less.
In Vivo Assay for Glucose Lowering
[0280] Oral glucose tolerance tests ("OGTT") have been in use in
humans since, at least, the 1930s, Pincus et al., Am. J. Med. Sci,
188: 782 (1934), and are routinely used in the diagnosis of human
diabetes, though not to evaluate the efficacy of therapeutic agents
in patients.
[0281] KK mice have been used to evaluate glitazones (Fujita et al.
Diabetes 32:804-810 (1983); Fujiwara et al., Diabetes 37: 1549-48
(1988); Izumi et al. Biopharm Durg. Dispos. 18:247-257 (1997)),
metformin (Reddi et al. Diabet. Metabl. 19:44-51 (1993)),
glucosidase inhibitors (Hamada et al. Jap. Pharmacol. Ther.
17:17-28 (1988); Matsuo et al. Am. J. Clin. Nutr. 55:314S-317S
(1992)), and the extra-pancreatic effects of sulfonylureas (Kameda
et al Arzenim. Forsch./Drug Res. 32:39044 (1982); Muller et al.
Horm. Metabl. Res. 28:469-487 (199)).
[0282] KK mice are derived from an inbred line first established by
Kondo et al. (Kondo et al. Bull. Exp. Anim. 6:107-112 (1957)). The
mice spontaneously develop a hereditary form of polygenic diabetes
that progresses to cause renal, retinal and neurological
complications analogous to those seen in human diabetic subjects,
but they do not require insulin or other medication for survival.
Another aspect of the invention is directed to the use of KK mice
to evaluate the effects of insulin secretagogue agents in the
context of an oral glucose tolerance test.
In Vivo Assay for Food Intake
[0283] The following screen may be used to evaluate the efficacy of
test compounds for inhibiting food intake in Sprague-Dawley rats
after an overnight fast.
[0284] Male Sprague-Dawley rats are individually housed and fed
powdered chow. They are maintained on a 12 hour light/dark cycle
and received food and water ad libitum. The animals are acclimated
to the vivarium for a period of one week before testing is
conducted. Testing is completed during the light portion of the
cycle.
[0285] To conduct the food intake efficacy screen, rats are
transferred to individual test cages without food the afternoon
prior to testing, and the rats are fasted overnight. After the
overnight fast, rats are dosed the following morning with vehicle
or test compounds. A known antagonist is dosed (3 mg/kg) as a
positive control, and a control group receives vehicle alone (no
compound). The test compounds are dosed at ranges between 0.1 and
100 mg/kg depending upon the compound. The standard vehicle is 0.5%
(w/v) methylcellulose in water and the standard route of
administration is oral. However, different vehicles and routes of
administration may be used to accommodate various compounds when
required. Food is provided to the rats 30 minutes after dosing and
an Oxymax automated food intake system (Columbus Instruments,
Columbus, Ohio) is started. Individual rat food intake is recorded
continuously at 10-minute intervals for a period of two hours. When
required, food intake is recorded manually using an electronic
scale; food is weighed every 30 minutes after food is provided up
to four hours after food is provided. Compound efficacy is
determined by comparing the food intake pattern of compound-treated
rats to vehicle and the standard positive control.
* * * * *