U.S. patent application number 14/628205 was filed with the patent office on 2015-06-18 for cyclic bridgehead ether dgat1 inhibitors.
This patent application is currently assigned to NOVARTIS AG. The applicant listed for this patent is Xin Chen, Yiping Ding, Rohit Duvadie, Yu Gai, Tyler Harrison, Jay Larrow, Qian Liu, Justin Mao, Sejal Patel, Jiong Ye, Frederic Zecri, Rui Zheng, Xuchun Zheng, Yizong Zhou. Invention is credited to Xin Chen, Yiping Ding, Rohit Duvadie, Yu Gai, Tyler Harrison, Jay Larrow, Qian Liu, Justin Mao, Sejal Patel, Jiong Ye, Frederic Zecri, Rui Zheng, Xuchun Zheng, Yizong Zhou.
Application Number | 20150166563 14/628205 |
Document ID | / |
Family ID | 48577190 |
Filed Date | 2015-06-18 |
United States Patent
Application |
20150166563 |
Kind Code |
A1 |
Patel; Sejal ; et
al. |
June 18, 2015 |
CYCLIC BRIDGEHEAD ETHER DGAT1 INHIBITORS
Abstract
The invention relates to compounds of formula (I): ##STR00001##
useful for treating disorders mediated by acyl coA-diacylglycerol
acyl transferase 1 (DGAT1), e.g. metabolic disorders. The invention
also provides methods of treating such disorders, and compounds and
compositions etc. for their treatment.
Inventors: |
Patel; Sejal; (Lexington,
MA) ; Mao; Justin; (North Reading, MA) ; Liu;
Qian; (Malden, MA) ; Zheng; Rui; (Chestnut
Hill, MA) ; Harrison; Tyler; (Somerville, MA)
; Duvadie; Rohit; (Arlington, MA) ; Chen; Xin;
(Lexington, MA) ; Zecri; Frederic; (Brookline,
MA) ; Larrow; Jay; (Cambridge, MA) ; Zheng;
Xuchun; (Changshu, CN) ; Zhou; Yizong;
(Changshu, CN) ; Ye; Jiong; (Changshu, CN)
; Ding; Yiping; (Changshu, CN) ; Gai; Yu;
(Changshu, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Patel; Sejal
Mao; Justin
Liu; Qian
Zheng; Rui
Harrison; Tyler
Duvadie; Rohit
Chen; Xin
Zecri; Frederic
Larrow; Jay
Zheng; Xuchun
Zhou; Yizong
Ye; Jiong
Ding; Yiping
Gai; Yu |
Lexington
North Reading
Malden
Chestnut Hill
Somerville
Arlington
Lexington
Brookline
Cambridge
Changshu
Changshu
Changshu
Changshu
Changshu |
MA
MA
MA
MA
MA
MA
MA
MA
MA |
US
US
US
US
US
US
US
US
US
CN
CN
CN
CN
CN |
|
|
Assignee: |
NOVARTIS AG
Basel
CH
|
Family ID: |
48577190 |
Appl. No.: |
14/628205 |
Filed: |
February 20, 2015 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
13871586 |
Apr 26, 2013 |
8993619 |
|
|
14628205 |
|
|
|
|
61639341 |
Apr 27, 2012 |
|
|
|
61787695 |
Mar 15, 2013 |
|
|
|
Current U.S.
Class: |
514/275 ;
514/337; 514/363; 514/364; 514/374; 544/331; 546/268.7; 546/269.1;
546/271.4; 548/138; 548/143; 548/236 |
Current CPC
Class: |
C07D 493/08 20130101;
A61P 25/28 20180101; A61K 45/06 20130101; A61P 3/10 20180101; A61K
31/4439 20130101; A61P 7/00 20180101; C07D 413/14 20130101; A61P
35/00 20180101; C07D 413/12 20130101; A61P 17/04 20180101; A61P
27/02 20180101; A61P 31/12 20180101; C07D 417/12 20130101; A61K
31/4245 20130101; A61K 31/433 20130101; A61P 3/06 20180101; A61P
1/18 20180101; A61P 3/04 20180101; A61P 1/16 20180101; A61K 31/422
20130101; C07D 417/14 20130101; A61K 31/506 20130101; A61P 3/00
20180101; A61P 43/00 20180101; A61P 3/08 20180101; A61K 31/4439
20130101; A61K 2300/00 20130101; A61K 31/506 20130101; A61K 2300/00
20130101; A61K 31/433 20130101; A61K 2300/00 20130101; A61K 31/4245
20130101; A61K 2300/00 20130101; A61K 31/422 20130101; A61K 2300/00
20130101 |
International
Class: |
C07D 493/08 20060101
C07D493/08; A61K 31/4439 20060101 A61K031/4439; C07D 417/14
20060101 C07D417/14; A61K 31/422 20060101 A61K031/422; C07D 417/12
20060101 C07D417/12; A61K 31/433 20060101 A61K031/433; C07D 413/12
20060101 C07D413/12; A61K 31/4245 20060101 A61K031/4245; C07D
413/14 20060101 C07D413/14; A61K 31/506 20060101 A61K031/506 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 15, 2013 |
CN |
PCT/CN2013/072735 |
Claims
1. A compound according to formula (I) or a salt or solvate
thereof: ##STR00131## Wherein p is 1, 2 or 3; X is O or CH.sub.2; Y
is O, CH.sub.2 or absent, wherein exactly one of X and Y is O;
Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.4 are each, independently, N or
CH; L is C(O) or absent; and A is a substituted oxazole, thiazole,
oxadiazole or thiadiazole substituted with at least one
C.sub.1-6alkyl, C.sub.3-7cycloalkyl or C.sub.1-6haloalkyl.
2. The compound according to claim 1, wherein the compound is of
formula (II) or a salt or solvate thereof: ##STR00132##
3. The compound according to claim 1, wherein the compound is of
formula (III) or a salt or solvate thereof: ##STR00133##
4. The compound according to claim 1, wherein the compound is of
formula (IV) or a salt or solvate thereof: ##STR00134##
5. The compound according to claim 1, or a salt or solvate thereof,
wherein p is 1.
6. The compound according to claim 1, or a salt or solvate thereof,
wherein p is 2.
7. The compound according to claim 1 or a salt or solvate thereof,
wherein Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.4 are all CH.
8. The compound according to claim 1, or a salt or solvate thereof,
wherein Z.sub.1 is N and Z.sub.2, Z.sub.3 and Z.sub.4 are each
CH.
9. The compound according to claim 1, or a salt or solvate thereof,
wherein Z.sub.2 is N and Z.sub.1, Z.sub.3 and Z.sub.4 are each
CH.
10. The compound according to claim 1, or a salt or solvate
thereof, wherein Z.sub.1 and Z.sub.2 are both N and Z.sub.3 and
Z.sub.4 are both CH.
11. The compound according to claim 1, or a salt or solvate
thereof, wherein L is C(O).
12. The compound according to claim 1, or a salt or solvate
thereof, wherein L is absent.
13. The compound according to claim 1, or a salt or solvate
thereof, wherein A is selected from: ##STR00135##
14. The compound according to claim 1, or a salt or solvate
thereof, wherein A is selected from: ##STR00136##
15. The compound according to claim 1, or a salt or solvate
thereof, wherein A is selected from ##STR00137##
16. The compound of claim 1, or a salt or solvate thereof, which
compound is selected from:
2-(4-(4'-((5-cyclobutyl-1,3,4-thiadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid;
2-(4-(4'-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid;
2-(4-(4-(5-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)pyridin-2-y-
l)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid;
2-(4-(4-(5-(2-ethyl-4-methyloxazole-5-carboxamido)pyridin-2-yl)phenyl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetic acid;
2-(4-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1'-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetic acid;
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid;
2-(1-(4'-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid;
2-(1-(4'-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)-[1,1'-biphen-
yl]-4-yl)-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid;
2-(1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-
-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid;
2-(1-(4-(5-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)pyridin-2-y-
l)phenyl)-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid;
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid;
3-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoic acid;
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)acetic acid; or a salt or solvate
thereof.
17. A pharmaceutical composition comprising one or more
pharmaceutically acceptable carriers and a therapeutically
effective amount of a compound of claim 1.
18. A combination, in particular a pharmaceutical combination,
comprising a therapeutically effective amount of the compound
according to claim 1 and a second therapeutically active agent.
19. A method for the treatment of a disease or condition mediated
by DGAT1 activity in a subject, wherein the method comprises
administering to the subject a therapeutically effective amount of
the compound according to claim 1.
20. The method of claim 19, in which the disease or condition is
selected from the group consisting of HCV, impaired glucose
tolerance, Type II diabetes or obesity.
21. A method of treating HCV, impaired glucose tolerance, Type II
diabetes or obesity comprising administering to a subject in need
thereof an effective amount of a composition comprising a compound
of claim 1.
Description
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional
Application No. 61/639,341 filed on Apr. 27, 2012, U.S. Provisional
Application No. 61/787,695 filed Mar. 15, 2013, and International
Application No. PCT/CN2013/072735 filed Mar. 15, 2013, the contents
of which are incorporated herein by reference in their
entireties.
TECHNICAL FIELD
[0002] This invention relates to compounds useful for treating
disorders mediated by acyl coA-diacylglycerol acyl transferase 1
(DGAT1), e.g. metabolic disorders. The invention also provides
methods of treating such disorders, and compounds and compositions
etc. for their treatment.
BACKGROUND ART
[0003] Although triglycerides (also known as "triacylglycerides")
are essential for normal physiology, excess triglyceride
accumulation results in obesity and, particularly when it occurs in
nonadipose tissues, is associated with insulin resistance. Obesity
increases the risk of many common and serious conditions, including
coronary heart disease, hypertension, dyslipidemia,
atherosclerosis, type-II diabetes, stroke, osteoarthritis,
restrictive pulmonary disease, sleep apnoea, certain types of
cancers and inflammatory disorders. The standard treatment for
obesity is calorific restriction and increase of physical exercise.
However, such approaches are rarely successful and pharmaceutical
treatments are required to correct these metabolic disorders.
[0004] A potential therapy for these conditions therefore involves
inhibiting triglyceride synthesis.
[0005] Diacylglycerol acyl-transference (DGAT) is an enzyme that
catalyzes the last step in triacylglycerol biosynthesis. DGAT
catalyzes the coupling of a 1,2-diacylglycerol with a fatty
acyl-CoA resulting in Coenzyme A and triacylglycerol. Two enzymes
that display DGAT activity have been identified: DGAT1 (acyl
coA-diacylglycerol acyl transferase 1) [Cases et al., Proc. Natl.
Acad. Sci. 1998, 95:13018-13023] and DGAT2 (acyl coA-diacylglycerol
acyl transferase 2) [Cases et al., J. Biol. Chem. 2001,
276:38870-38876].
[0006] DGAT1 and DGAT2 do not share significant protein sequence
homology. Importantly, however, DGAT1 knockout mice are protected
from high fat diet-induced weight gain and insulin resistance
[Smith et al., Nature Genetics 2000, 25:87-90]. The phenotype of
the DGAT1 knockout mice suggests that DGAT1 inhibitors would be
useful for the treatment of obesity and obesity-associated
complications [Smith et al., Nature Genetics 2000, 25:87-90].
[0007] There is therefore a need for compounds which inhibit the
activity of DGAT1.
DISCLOSURE OF THE INVENTION
[0008] The inventors have found compounds of formula (I) that are
useful for inhibiting the activity of DGAT1.
[0009] Accordingly, in a first embodiment of the invention, there
is provided a compound of formula (I) or a salt or solvate
thereof:
##STR00002##
wherein p is 1, 2 or 3; X is O or CH.sub.2; Y is O, CH.sub.2 or
absent, wherein exactly one of X and Y is O; Z.sub.1, Z.sub.2,
Z.sub.3 and Z.sub.4 are each, independently, N or CH; L is C(O) or
absent; and A is a substituted oxazole, thiazole, oxadiazole or
thiadiazole substituted with at least one C.sub.1-6alkyl,
C.sub.3-7cycloalkyl or C.sub.1-6haloalkyl.
[0010] In a second embodiment according to the first embodiment,
the invention is a compound of formula (I) wherein p is 1.
[0011] In a third embodiment according to the first embodiment, the
invention is a compound of formula (I) wherein p is 2.
[0012] In a fourth embodiment, the invention is a compound of
formula (II) or a salt or solvate thereof:
##STR00003##
[0013] In fifth embodiment, the invention is a compound of formula
(III) or a salt or solvate thereof:
##STR00004##
[0014] In sixth embodiment, the invention is a compound of formula
(IV) or a salt or solvate thereof:
##STR00005##
[0015] In a seventh embodiment according to any one of the first to
sixth embodiments, the invention is a compound according formula
(I), (II), (III) and (IV) wherein the variables Z.sub.1, Z.sub.2,
Z.sub.3 and Z.sub.4 are all CH.
[0016] In an eighth embodiment according to any one of the first to
sixth embodiments, the invention is a compound according formula
(I), (II), (III) and (IV) wherein the variable Z.sub.1 is N and the
variables Z.sub.2, Z.sub.3 and Z.sub.4 are each CH.
[0017] In a ninth embodiment according to any one of the first to
sixth embodiments, the invention is a compound according formula
(I), (II), (III) and (IV) wherein the variable Z.sub.2 is N and the
variables Z.sub.1, Z.sub.3 and Z.sub.4 are each CH.
[0018] In a tenth embodiment according to any one of the first to
sixth embodiments, the invention is a compound according formula
(I), (II), (III) and (IV) wherein the variables Z.sub.1 and Z.sub.2
are both N and the variables Z.sub.3 and Z.sub.4 are both CH.
[0019] In an eleventh embodiment according to any one of the first
to tenth embodiments, the invention is a compound according formula
(I), (II), (III) and (IV) wherein the variable L is C(O).
[0020] In a twelfth embodiment according to any one of the first to
tenth embodiments, the invention is a compound according formula
(I), (II), (III) and (IV) wherein the variable wherein L is
absent.
[0021] In a thirteenth embodiment according to any one of the first
to twelfth embodiments, the invention is a compound according
formula (I), (II), (III) and (IV) wherein the variable A is
selected from:
##STR00006##
[0022] In an fourteenth embodiment according to any one of the
first to tenth and twelfth embodiments, the invention is a compound
according to formula (I), (II), (III) or (IV) or a salt or solvate
thereof, wherein L is absent and A is selected from:
##STR00007##
[0023] In an fifteenth embodiment according to any one of the first
to eleventh embodiments, the invention is a compound of formula
(I), (II), (III) or (IV) or a salt or solvate thereof, wherein L is
C(O) and A is selected from
##STR00008##
[0024] In a sixteenth embodiment according to any one of the first
to fifteenth embodiments, the invention is a pharmaceutical
composition comprising one or more pharmaceutically acceptable
carriers and a therapeutically effective amount of a compound
according to formula (I), (II), (III) and (IV).
[0025] In a seventeenth embodiment according to any one of the
first to fifteenth embodiments, the invention is a combination, in
particular a pharmaceutical combination, comprising a
therapeutically effective amount of the compound according to
formula (I), (II), (III) and (IV) and a second therapeutically
active agent.
[0026] In an eighteenth embodiment according to any one of the
first to fifteenth embodiments, the invention is a method for the
treatment of a disease or condition mediated by DGAT1 activity in a
subject, wherein the method comprises administering to the subject
a therapeutically effective amount of the compound according to
formula (I), (II), (III) and (IV).
[0027] In a nineteenth embodiment according to any one of the first
to fifteenth and eighteenth embodiments, the disease or condition
is selected from the group consisting of HCV, impaired glucose
tolerance, Type II diabetes or obesity.
[0028] In a twentieth embodiment according to any one of the first
to fifteenth embodiments, the invention is a method of treating
HCV, impaired glucose tolerance, Type II diabetes or obesity
comprising administering to a subject in need thereof an effective
amount of a composition comprising a compound of to formula (I),
(II), (III) and (IV).
[0029] In a twentyfirst embodiment according to any one of the
first to fifteenth embodiments, the invention is a compound
according to formula (I), (II), (III) and (IV), for use as a
medicament.
[0030] In a twentysecond embodiment according to any one of the
first to fifteenth embodiments, the invention is the use of a
compound according to formula (I), (II), (III) and (IV), in the
manufacture of a medicament for the treatment of a disease or
condition mediated by DGAT1 activity.
[0031] In a twentythird embodiment according to any one of the
first to fifteenth embodiments, the invention is the use of a
compound to formula (I), (II), (III) and (IV), for the treatment of
HCV, impaired glucose tolerance, Type II diabetes or obesity.
[0032] In a twentyfourth embodiment according to any one of the
first to fifteenth embodiments, the invention is a method for the
prevention, delay of progression or treatment of a disease
exacerbated by inadequate phosphatidylcholine production,
comprising: administering to a warm-blooded animal in need thereof
a therapeutically effective amount of a DGAT1 inhibitor of formula
(I), (II), (III) or (IV). In an exemplary embodiment, the
warm-blooded animal is a human.
[0033] In a twentyfifth embodiment according to any one of the
first to fifteenth embodiments, the invention is use of a DGAT1
inhibitor of formula (I), (II), (III) or (IV) for the preparation
of a pharmaceutical composition for the treatment of a disorder or
disease exacerbated by inadequate phosphatidylcholine production in
a subject mediated by the inhibition of DGAT1.
[0034] In a twentysixth embodiment according to any one of the
first to fifteenth embodiments, the invention is a DGAT1 inhibitor
of formula (I), (II), (III), (IV), or a pharmaceutically acceptable
salt or ester thereof; for use in the prevention, delay of
progression or treatment of a disease or condition which is
selected from chylomicronemia 5 syndrome, familial chylomicronemia
syndrome, and Type V hyperlipoproteinemia.
[0035] In a twentyseventh embodiment according to any one of the
first to fifteenth embodiments, the invention is a DGAT1 inhibitor
of formula (I), (II), (III) or (IV), or a pharmaceutically
acceptable salt or ester thereof, for use in the reduction of
postprandial triglyceride levels in patients suffering from a
disease or condition which is selected from chylomicronemia
syndrome, familial chylomicronemia syndrome, and Type V
hyperlipoproteinemia.
[0036] In a twentyeighth embodiment according to any one of the
first to fifteenth embodiments, the invention is a DGAT1 inhibitor
of formula (I), (II), (III) or (IV), or a pharmaceutically
acceptable salt or ester thereof, for use in the prevention, delay
of progression or treatment of pancreatitis in patients suffering
from a disease or condition which is selected from chylomicronemia
syndrome, familial chylomicronemia syndrome, and Type V
hyperlipoproteinemia.
[0037] In a twentyninth embodiment according to any one of the
first to fifteenth embodiments, the invention is a DGAT1 inhibitor
of formula (I), (II), (III) or (IV), or a pharmaceutically
acceptable salt or ester thereof, for use in the prevention, delay
of progression or treatment of a symptom selected from recurrent
episodes of pancreatitis, deposition of triglycerides in the skin
in the form of eruptive xanthomas, hepatosplenomegaly, milky white
triglyceride in the blood vessels in the back of the eye (lipemia
retinalis), and mild neuro-cognitive deficits.
[0038] In a thirtieth embodiment according to the first embodiment,
the invention is a compound according formula (I), (II), (III) and
(IV) wherein the compound is selected from at least one of: [0039]
2-(4-(4'-((5-cyclobutyl-1,3,4-thiadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid; [0040]
2-(4-(4'-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid; [0041]
2-(4-(4-(5-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)pyridin-2-y-
l)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid; [0042]
2-(4-(4-(5-(2-ethyl-4-methyloxazole-5-carboxamido)pyridin-2-yl)phenyl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetic acid; [0043]
2-(4-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1'-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetic acid; [0044]
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid; [0045]
2-(1-(4'-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid; [0046]
2-(1-(4'-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)-[1,1'-biphen-
yl]-4-yl)-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid; [0047]
2-(1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-
-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid; [0048]
2-(1-(4-(5-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)pyridin-2-y-
l)phenyl)-2-oxabicyclo[2.2.2]octan-4-yl)acetic acid; [0049]
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid; [0050]
3-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoic acid; [0051]
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)acetic acid; or a salt or solvate
thereof.
Compounds of Formula (I)-(IV) etc. and Derivatives Thereof
[0052] As used herein, the terms "compounds of the invention" and
"compound of formula (I)" etc. include pharmaceutically acceptable
derivatives thereof and polymorphs, isomers and isotopically
labelled variants thereof. Furthermore, the term "compounds of the
invention" and "compound of formula (I)" etc include compounds of
formula (II), (111) and (IV), and the embodiments thereof disclosed
herein.
Pharmaceutically Acceptable Derivatives
[0053] The term "pharmaceutically acceptable derivative" includes
any pharmaceutically acceptable salt, solvate, hydrate or prodrug
of a compound of formula (I). In one embodiment, the
pharmaceutically acceptable derivatives are pharmaceutically
acceptable salts, solvates or hydrates of a compound of formula
(I).
Pharmaceutically Acceptable Salts
[0054] As used herein, the terms "salt" or "salts" refers to an
acid addition or base addition salt of a compound of the invention.
"Salts" include in particular "pharmaceutical acceptable salts".
The term "pharmaceutically acceptable salts" refers to salts that
retain the biological effectiveness and properties of the compounds
of this invention and, which typically are not biologically or
otherwise undesirable. In many cases, the compounds of the present
invention are capable of forming acid and/or base salts by virtue
of the presence of amino and/or carboxyl groups or groups similar
thereto.
[0055] Pharmaceutically acceptable acid addition salts can be
formed with inorganic acids and organic acids, e.g., acetate,
aspartate, benzoate, besylate, bromide/hydrobromide,
bicarbonate/carbonate, bisulfate/sulfate, camphorsulfonate,
chloride/hydrochloride, chlortheophyllonate, citrate,
ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate,
hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate,
laurylsulfate, malate, maleate, malonate, mandelate, mesylate,
methylsulphate, naphthoate, napsylate, nicotinate, nitrate,
octadecanoate, oleate, oxalate, palmitate, pamoate,
phosphate/hydrogen phosphate/dihydrogen phosphate,
polygalacturonate, propionate, stearate, succinate,
sulfosalicylate, tartrate, tosylate and trifluoroacetate salts.
[0056] Inorganic acids from which salts can be derived include, for
example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, and the like.
[0057] Organic acids from which salts can be derived include, for
example, acetic acid, propionic acid, glycolic acid, oxalic acid,
maleic acid, malonic acid, succinic acid, fumaric acid, tartaric
acid, citric acid, benzoic acid, mandelic acid, methanesulfonic
acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic
acid, and the like. Pharmaceutically acceptable base addition salts
can be formed with inorganic and organic bases.
[0058] Inorganic bases from which salts can be derived include, for
example, ammonium salts and metals from columns I to XII of the
periodic table. In certain embodiments, the salts are derived from
sodium, potassium, ammonium, calcium, magnesium, iron, silver,
zinc, and copper; particularly suitable salts include ammonium,
potassium, sodium, calcium and magnesium salts.
[0059] Organic bases from which salts can be derived include, for
example, primary, secondary, and tertiary amines, substituted
amines including naturally occurring substituted amines, cyclic
amines, basic ion exchange resins, and the like. Certain organic
amines include isopropylamine, benzathine, cholinate,
diethanolamine, diethylamine, lysine, meglumine, piperazine and
tromethamine.
[0060] The pharmaceutically acceptable salts of the present
invention can be synthesized from a basic or acidic moiety, by
conventional chemical methods. Generally, such salts can be
prepared by reacting free acid forms of these compounds with a
stoichiometric amount of the appropriate base (such as Na, Ca, Mg,
or K hydroxide, carbonate, bicarbonate or the like), or by reacting
free base forms of these compounds with a stoichiometric amount of
the appropriate acid. Such reactions are typically carried out in
water or in an organic solvent, or in a mixture of the two.
Generally, use of non-aqueous media like ether, ethyl acetate,
ethanol, isopropanol, or acetonitrile is desirable, where
practicable. Lists of additional suitable salts can be found, e.g.,
in "Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing
Company, Easton, Pa., (1985); and in "Handbook of Pharmaceutical
Salts: Properties, Selection, and Use" by Stahl and Wermuth
(Wiley-VCH, Weinheim, Germany, 2002).
Solvates & Hydrates
[0061] The compounds of the invention may exist in both unsolvated
and solvated forms. The term "solvate" includes molecular complexes
comprising a compound of the invention and one or more
pharmaceutically acceptable solvent molecules such as water or
C.sub.1-6 alcohols, e.g. ethanol. The term "hydrate" means a
"solvate" where the solvent is water.
Prodrugs
[0062] Furthermore, the compounds of the present invention,
including their salts, can also be obtained in the form of their
hydrates, or include other solvents used for their crystallization.
The compounds of the present invention may inherently or by design
form solvates with pharmaceutically acceptable solvents (including
water); therefore, it is intended that the invention embrace both
solvated and unsolvated forms. The term "solvate" refers to a
molecular complex of a compound of the present invention (including
pharmaceutically acceptable salts thereof) with one or more solvent
molecules. Such solvent molecules are those commonly used in the
pharmaceutical art, which are known to be innocuous to the
recipient, e.g., water, ethanol, and the like. The term "hydrate"
refers to the complex where the solvent molecule is water.
[0063] The compounds of the present invention, including salts,
hydrates and solvates thereof, may inherently or by design form
polymorphs.
Description of Isomeric Forms and Separation Methods
[0064] Any asymmetric atom (e.g., carbon or the like) of the
compound(s) of the present invention can be present in racemic or
enantiomerically enriched, for example the (R)-, (S)- or
(R,S)-configuration. In certain embodiments, each asymmetric atom
has at least 50% enantiomeric excess, at least 60% enantiomeric
excess, at least 70% enantiomeric excess, at least 80% enantiomeric
excess, at least 90% enantiomeric excess, at least 95% enantiomeric
excess, or at least 99% enantiomeric excess in the (R)- or
(S)-configuration. Substituents at atoms with unsaturated double
bonds may, if possible, be present in cis-(Z)- or
trans-(E)-form.
[0065] Accordingly, as used herein a compound of the present
invention can be in the form of one of the possible isomers,
rotamers, atropisomers, tautomers or mixtures thereof, for example,
as substantially pure geometric (cis or trans) isomers,
diastereomers, optical isomers (antipodes), racemates or mixtures
thereof.
[0066] Any resulting mixtures of isomers can be separated on the
basis of the physicochemical differences of the constituents, into
the pure or substantially pure geometric or optical isomers,
diastereomers, racemates, for example, by chromatography and/or
fractional crystallization.
[0067] Any resulting racemates of final products or intermediates
can be resolved into the optical antipodes by known methods, e.g.,
by separation of the diastereomeric salts thereof, obtained with an
optically active acid or base, and liberating the optically active
acidic or basic compound. In particular, a basic moiety may thus be
employed to resolve the compounds of the present invention into
their optical antipodes, e.g., by fractional crystallization of a
salt formed with an optically active acid, e.g., tartaric acid,
dibenzoyl tartaric acid, diacetyl tartaric acid, di-O,O'-p-toluoyl
tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic
acid. Racemic products can also be resolved by chiral
chromatography, e.g., high pressure liquid chromatography (HPLC)
using a chiral adsorbent.
Amorphous & Crystalline Forms
[0068] The compounds of the invention may exist in solid states
from amorphous through to crystalline forms. All such solid forms
are included within the invention.
Isomeric Forms
[0069] As used herein, the term "an optical isomer" or "a
stereoisomer" refers to any of the various stereo isomeric
configurations which may exist for a given compound of the present
invention and includes geometric isomers. It is understood that a
substituent may be attached at a chiral center of a carbon atom.
The term "chiral" refers to molecules which have the property of
non-superimposability on their mirror image partner, while the term
"achiral" refers to molecules which are superimposable on their
mirror image partner. Therefore, the invention includes
enantiomers, diastereomers or racemates of the compound.
"Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term is used to
designate a racemic mixture where appropriate.
[0070] "Diastereoisomers" are stereoisomers that have at least two
asymmetric atoms, but which are not mirror-images of each other.
The absolute stereochemistry is specified according to the
Cahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer
the stereochemistry at each chiral carbon may be specified by
either R or S. Resolved compounds whose absolute configuration is
unknown can be designated (+) or (-) depending on the direction
(dextro- or levorotatory) which they rotate plane polarized light
at the wavelength of the sodium D line. Certain compounds described
herein contain one or more asymmetric centers or axes and may thus
give rise to enantiomers, diastereomers, and other stereoisomeric
forms that may be defined, in terms of absolute stereochemistry, as
(R)- or (S)-.
[0071] Depending on the choice of the starting materials and
procedures, the compounds can be present in the form of one of the
possible isomers or as mixtures thereof, for example as pure
optical isomers, or as isomer mixtures, such as racemates and
diastereoisomer mixtures, depending on the number of asymmetric
carbon atoms. The present invention is meant to include all such
possible isomers, including racemic mixtures, diasteriomeric
mixtures and optically pure forms. Optically active (R)- and
(S)-isomers may be prepared using chiral synthons or chiral
reagents, or resolved using conventional techniques. If the
compound contains a double bond, the substituent may be E or Z
configuration. If the compound contains a disubstituted cycloalkyl,
the cycloalkyl substituent may have a cis- or trans-configuration.
All tautomeric forms are also intended to be included.
Isotopic Labeling
[0072] Any formula given herein is also intended to represent
unlabeled forms as well as isotopically labelled forms of the
compounds. Isotopically labelled compounds have structures depicted
by the formulas given herein except that one or more atoms are
replaced by an atom having a selected atomic mass or mass number.
Examples of isotopes that can be incorporated into compounds of the
invention include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorous, fluorine, and chlorine, such as .sup.2H, .sup.3H,
.sup.11C, .sup.13C, .sup.14C, .sup.15N, .sup.18F .sup.31P,
.sup.32P, .sup.35S, .sup.36Cl, .sup.125I respectively. The
invention includes various isotopically labelled compounds as
defined herein, for example those into which radioactive isotopes,
such as .sup.3H and .sup.14C, or those into which non-radioactive
isotopes, such as .sup.2H and .sup.13C are present. Such
isotopically labelled compounds are useful in metabolic studies
(with .sup.14C), reaction kinetic studies (with, for example
.sup.2H or .sup.3H), detection or imaging techniques, such as
positron emission tomography (PET) or single-photon emission
computed tomography (SPECT) including drug or substrate tissue
distribution assays, or in radioactive treatment of patients. In
particular, an .sup.18F or labelled compound may be particularly
desirable for PET or SPECT studies. Isotopically-labelled compounds
of formula (I) can generally be prepared by conventional techniques
known to those skilled in the art or by processes analogous to
those described in the accompanying Examples and Preparations using
an appropriate isotopically-labelled reagents in place of the
non-labelled reagent previously employed.
[0073] Further, substitution with heavier isotopes, particularly
deuterium (i.e., .sup.2H or D) may afford certain therapeutic
advantages resulting from greater metabolic stability, for example
increased in vivo half-life or reduced dosage requirements or an
improvement in therapeutic index. It is understood that deuterium
in this context is regarded as a substituent of a compound of the
formula (I). The concentration of such a heavier isotope,
specifically deuterium, may be defined by the isotopic enrichment
factor. The term "isotopic enrichment factor" as used herein means
the ratio between the isotopic abundance and the natural abundance
of a specified isotope. If a substituent in a compound of this
invention is denoted deuterium, such compound has an isotopic
enrichment factor for each designated deuterium atom of at least
3500 (52.5% deuterium incorporation at each designated deuterium
atom), at least 4000 (60% deuterium incorporation), at least 4500
(67.5% deuterium incorporation), at least 5000 (75% deuterium
incorporation), at least 5500 (82.5% deuterium incorporation), at
least 6000 (90% deuterium incorporation), at least 6333.3 (95%
deuterium incorporation), at least 6466.7 (97% deuterium
incorporation), at least 6600 (99% deuterium incorporation), or at
least 6633.3 (99.5% deuterium incorporation).
[0074] Pharmaceutically acceptable solvates in accordance with the
invention include those wherein the solvent of crystallization may
be isotopically substituted, e.g. D.sub.2O, d.sub.6-acetone,
d.sub.6-DMSO.
Co-Crystals
[0075] Compounds of the invention, i.e. compounds of formula (I)
that contain groups capable of acting as donors and/or acceptors
for hydrogen bonds may be capable of forming co-crystals with
suitable co-crystal formers. These co-crystals may be prepared from
compounds of formula (I) by known co-crystal forming procedures.
Such procedures include grinding, heating, co-subliming,
co-melting, or contacting in solution compounds of formula (I) with
the co-crystal former under crystallization conditions and
isolating co-crystals thereby formed. Suitable co-crystal formers
include those described in WO 2004/078163. Hence the invention
further provides co-crystals comprising a compound of formula
(I).
Treatment of Diseases and Conditions
[0076] Compounds of formula (I) have been found to be inhibitors of
DGAT1.
[0077] The invention provides a compound of formula (I) for use in
therapy. The invention further provides a pharmaceutical
composition comprising a compound of formula (I) in combination
with a pharmaceutically acceptable excipient.
[0078] The invention further provides a method for the treatment of
a disease or condition mediated by DGAT1, comprising the step of
administering a therapeutically effective amount of a compound of
formula (I) to a patient. The invention also provides the use of a
compound of formula (I) in the manufacture of a medicament for the
treatment of a disease or condition mediated by DGAT1. The
invention also provides a compound of formula (I) for use in
treating a disease or condition mediated by DGAT1.
[0079] The invention also provides a crystal of DGAT1 and a
compound of formula (I). Such crystals can be used for X-ray
diffraction studies of DGAT1 inhibition, e.g. to provide atomic
structural information in order to aid rational design of further
DGAT1 inhibitors.
[0080] The DGAT1 inhibitory activity of the compounds of the
invention may be demonstrated by the DGAT1 assay disclosed herein
(see "DGAT1 Inhibition Assay"). Preferred compounds of the
invention have an IC.sub.50 in the DGAT1 Inhibition Assay of
<100 .mu.M, in one embodiment <10 .mu.M, in another
embodiment <1 .mu.M, in another embodiment <100 nM, and in
another embodiment <10 nM.
Diseases and Conditions Mediated by DGAT1
[0081] The invention is useful for the treatment of a disease or
condition mediated by DGAT1. Diseases and conditions mediated by
DGAT1 include: metabolic disorders such as obesity, diabetes (e.g.
Type II diabetes), anorexia nervosa, bulimia, cachexia, syndrome X,
insulin resistance, glucose tolerance, hypoglycemia, hyperglycemia,
hyperuricemia, hyperinsulinemia, hypercholesterolemia,
hyperlipidemia, dyslipidemia, mixed dyslipidemia,
hypertriglyceridemia, pancreatitis, and nonalcoholic fatty liver
disease; cardiovascular diseases, such as atherosclerosis,
arteriosclerosis, acute heart failure, congestive heart failure,
coronary artery disease, cardiomyopathy, myocardial infarction,
angina pectoris, hypertension, hypotension, stroke, ischemia,
myocardial ischemia, ischemic reperfusion injury, aneurysm,
restenosis, and vascular stenosis; neoplastic diseases, such as
solid tumors, skin cancer, melanoma, lymphoma, and endothelial
cancers, for example, breast cancer, lung cancer, colorectal
cancer, stomach cancer, other cancers of the gastrointestinal tract
(e.g. esophageal cancer and pancreatic cancer), prostate cancer,
kidney cancer, liver cancer, bladder cancer, cervical cancer,
uterine cancer, testicular cancer, and ovarian cancer;
dermatological conditions, such as acne vulgaris; hepatitis C virus
(HCV); pathogens that target lipid droplets (e.g. dengue and
chlamydia); and infectious agents which require lipid droplets
and/or triglycerides in their lifecycle.
[0082] One embodiment, the disease or condition mediated by DGAT 1
is impaired glucose tolerance (IGT), Type II diabetes or
obesity.
[0083] As used herein a patient is suffering from "obesity" if the
patient exhibits at least one of: [0084] a body mass index (BMI),
i.e. the patient's mass (in kg) divided by the square of the
patient's height (in m), of 30 or more; [0085] an absolute waist
circumference of >102 cm in men or >88 cm in women; [0086] a
waist-to-hip ratio >0.9 in men or >0.85 in women; or [0087] a
percent body fat >25% in men or >30% in women.
[0088] As used herein a patient is suffering from "Type II
diabetes" if they meet the World Health Organization criteria for
Diabetes diagnosis (Definition and diagnosis of diabetes mellitus
and intermediate hyperglycemia, WHO, 2006), i.e. the patient
exhibits at least one of: [0089] a fasting plasma glucose
.gtoreq.7.0 mmol/l (126 mg/dl); or [0090] a venous plasma glucose
.gtoreq.11.1 mmol/l (200 mg/dl) 2 hours after ingestion of 75 g
oral glucose load.
[0091] As used herein a patient is suffering from "IGT" if they
meet the World Health Organization criteria for IGT diagnosis
(Definition and diagnosis of diabetes mellitus and intermediate
hyperglycemia, WHO, 2006), i.e. the patient exhibits both of:
[0092] a fasting plasma glucose <7.0 mmol/l (126 mg/dl); and
[0093] a venous plasma glucose .gtoreq.7.8 and <11.1 mmol/l (200
mg/dl) 2 hours after ingestion of 75 g oral glucose load.
[0094] In yet another aspect, the invention is useful as an
anorectic.
[0095] In one embodiment, the disease or condition mediated by DGAT
1 is HCV (Harris C, Hernandez C, Carpentier A, Kaehlcke K,
Rosenberg A R, Farese R V Jr, Ott M Efficient hepatitis C virus
particle formation requires diacylglycerol acyltransferase-1.
Herker E, Nat Med. 2010 November; 16(11):1295-8. & Charles
Harris, Eva Herker, Robert V. Farese Jr., Melanie Ott, The Journal
of Biological Chemistry, 286, 42615-42625.)
[0096] In another embodiment, the disease or condition mediated by
DGAT1 is myocardial ischemia Stanley, W. C., Expert opinion in
Investig. Drugs; 11(5): 615-629, 2002; and Dyck, J. R. and
Lopaschuk, G. D., J. Mol. Cell. Cardiol. 34(9): 1099-1109,
2002].
[0097] In one embodiment, increasing cellular phosphatidyl choline
is achieved by DGAT1 inhibition and used as an approach to the
therapeutic increase of phosphatidylcholine in plasma lipoproteins
and intestinal epithelium (Kent C, Biochim. Biophys. Acta, 1733:
53-66, 2005; Coleman R A, Prog. Lipid Res., 34: 134-176, 2004; Goni
F M, et al. Prog. Lipid Res. 38: 1-48, 1999; Jenkins G M, et al.,
Cell. Mol. Life Sci. 62: 2305-2316, 2005; Becker K P, et al. Cell
Mol. Life Sci. 62: 1448-1461, 2005; Kruit J K, et al., World J.
Gastroenterol., 12: 6429-6439, 2006; Lewis G F, Curr. Opin.
Cardiol., 21: 345-352, 2006; Ehehalt R, Scand. J. of
Gastroenterology, 39: 737-742; Stremmel W, Gut, 54: 966-971, 2005;
Treede I, J. Biol. Chem., 282: 27155-27164, 2007; Cases et al,
Proc. Natl. Acad. Sci. 95:13018-13023, 1998; Cases et al, J. Biol.
Chem. 276:38870-38876, 2001; and Smith et al, Nature Genetics
25:87-90, 2000).
[0098] In another embodiment, the present invention relates to the
use of a DGAT1 inhibitor, or a pharmaceutically acceptable salt or
ester thereof, for the treatment of, or the prevention, delay of
progression, or treatment of a disease or condition which is
selected from chylomicronemia syndrome, familial chylomicronemia
syndrome and Type V hyperlipoproteinemia. The present invention
further relates to the use of a pharmaceutical composition
comprising a DGAT1 inhibitor, or a pharmaceutically acceptable salt
or ester thereof, for the prevention, delay of progression, or
treatment of a disease or condition which is selected from
chylomicronemia syndrome, familial chylomicronemia syndrome and
Type V hyperlipoproteinemia.
[0099] Hyperlipidemia, or the presence of elevated levels of lipids
in the bloodstream, can take the form of hypercholesterolemia
(elevated cholesterol), hypertriglyceridemia (elevated
triglyceride) or a combination of the two. Hypercholesterolemia,
which can further be subdivided, is typically associated with
increased risk of atherosclerosis cardiovascular disease.
Hypertriglyceridemia occurs when the body's production or intake of
triglyceride exceeds the body's ability to metabolize or remove the
triglyceride from the bloodstream. The most severe form of
hypertriglyceridemia is chylomicronemia (also called
hyperchylomicronemia), and is associated with an increased risk of
pancreatitis. Chylomicrons are lipoprotein particles that carry
absorbed dietary fat from the gut to other body tissues via the
bloodstream, and are typically present only during meal times.
Chylomicronemia is defined as having the presence of chylomicrons
in the bloodstream during times of fasting, and is typically
associated with total plasma triglyceride levels above 1000
mg/dL.
[0100] The chylomicronemia syndrome refers to a set of clinical
complications associated with high chylomicron levels. Typically,
patients with the chylomicronemia syndrome have markedly elevated
fasting triglyceride levels (1000-2000 mg/dL) with profound
excursions (up to 5000 mg/dL and higher) following oral fat intake.
The massively elevated plasma triglyceride levels are associated
with a number of clinical findings and complications including
recurrent episodes of pancreatitis, deposition of triglycerides in
the skin in the form of eruptive xanthomas, epatosplenomegaly, a
milky pink appearance of the blood vessels in the back of the eye
(lipemia retinalis), and mild neuro-cognitive deficits.
[0101] The chylomicronemia syndrome can be further subdivided into
two groups based on ultracentrifugation of lipoprotein species (see
"A system for phenotyping hyperlipoproteinemia", Fredrickson D. S.,
Lees R. S. Circulation, 1965 March; 31, pp. 321-327).
[0102] Fredrickson classification Type I, also known as the
familial chylomicronemia syndrome (FCS), patients have accumulation
of only chylomicrons in the bloodstream whereas Fredrickson
classification Type V, also known as Type V hyperlipoproteinemia,
patients have accumulation of both chylomicrons and very low
density lipoproteins (VLDL) in the bloodstream.
[0103] The familial chylomicronemia syndrome (FCS or Type I
hyperlipoproteinemia) is caused by a homozygous or compound
heterozygous defect in the clearance of chylomicrons from the
bloodstream. The most common cause of FCS is a defect in
lipoprotein lipase (LPL), the protein that hydrolyzes triglycerides
carried on chylomicrons. Other causes of FCS include defects in
apolipoprotein CII (apoCII, a co-activator of LPL) or
glycosylphosphatidylinositol-anchored high-density
lipoprotein-binding protein 1 (GPIHBP1, an anchoring protein of
LPL).
[0104] Type I patients are usually identified by early onset as
youth of hypertriglyceridemia and pancreatitis. Thus, patients with
FCS typically present in childhood with massively elevated
triglyceride levels (>2,000 mg/dL), and recurrent bouts of
abdominal pain due to pancreatitis. Into adulthood, the
triglyceride levels remain elevated, and patients typically
experience multiple episodes of abdominal pain and pancreatitis,
which can result in hospitalization and death.
[0105] Patients also experience other manifestations including
eruptive xanthomas, lipemia retinalis, hepatosplenomegaly, and mild
neuro-cognitive deficits. The main therapeutic goal in FCS
treatment is to prevent or treat pancreatitis via the reduction of
triglycerides.
[0106] Unfortunately, standard lipid-lowering therapies, such as
fibrates, omega-3 fatty acids, statins and nicotinic acid
derivatives (niacin), are not effective in lowering triglycerides
in patients with FCS. Therefore, the standard of care therapy for
FCS patients is a very low fat diet 10% by calories), something
which is very difficult to stay compliant with throughout a
lifetime [The Familial Chylomicronemia Syndrome. Santamarina-Fojo
S. Lipid Disorders 1998. 27(3): 551-567].
[0107] Another approach to treat FCS that is under investigation is
gene therapy using a replication-deficient Adeno-Associated Viral
vector to deliver a naturally-occurring, "beneficial" variant of
LPL (Glybera.RTM.) intramuscularly. However this treatment is only
transiently effective and requires immunosuppression with
mycophenolate, cyclosporine, and steroids [Alipogene tiparvovec,
and adeno-associated virus encoding the Ser(447)X variant of human
lipoprotein lipase gene for the treatment of patients with
lipoprotein lipase deficiency. Burnett J R., Hooper A J. Curr Opin
Mol Ther 2009. 6:681-691].
[0108] At present there is thus no effective pharmacotherapy for
treating FCS and there is thus a need for new methods of treating
familial chylomicronemia syndrome (FCS), also known as Type I
hyperlipoproteinemia.
[0109] Type V hyperlipoproteinemia patients represent a second
group at risk for the chylomicronemia syndrome and are usually
diagnosed by severe hypertriglyceridemia as adults. This is a
heterogeneous group at the extreme end of a spectrum of
multifactorial hypertriglyceridemia.
[0110] Patients with Type V hyperlipoproteinemia generally have
both an underlying genetic cause and one or more acquired causes of
hypertriglyceridemia. The underlying genetic causes include well
characterized dyslipidemia such as familial combined hyperlipidemia
(Type IIA), dysbetalipoproteinemia (Type III) and familial
hypertriglyceridemia (Type VI), and a group of less well
characterized dyslipidemias (e.g. heterozygous LPL deficiency,
defects in apoA & apoC genes, defects in fatty acid binding and
transport proteins).
[0111] Acquired causes of hypertriglyceridemia include comorbid
diseases (e.g. type 2 diabetes, obesity, insulin resistance,
lipodystrophy, hypothyroidism), medications (e.g. beta blockers,
thiazide diuretics, estrogen, glucocorticoids, transplant
medications), and other factors (e.g. pregnancy, alcohol
intake).
[0112] The primary goal of therapy in Type V patients is to reduce
the triglyceride levels, and therefore reduce the risk of
pancreatitis. Most patients can be successfully treated by
addressing the underlying acquired cause(s) of the elevated
triglycerides, such as reducing the amount of dietary fat intake,
treating uncontrolled co-morbid diseases such as T2DM (Type 2
diabetes mellitus), discontinuing offending medications, and
initiating lipid lowering medications such as fibrates, omega-3
fatty acids, or nicotinic acid derivatives (niacin)
[Chylomicronemia Syndrome. Chait A., Brunzell J. Adv Intern Med 5
1992. 37:249-73.].
[0113] Despite optimal therapy, some Type V patients continue to
have elevated triglyceride levels. There is thus a need for new
methods of treating Type V hyperlipoproteinemia.
Therapeutic Definitions
[0114] As used herein, "treatment" includes curative and
prophylactic treatment. As used herein, a "patient" means an
animal, preferably a mammal, preferably a human, in need of
treatment.
[0115] The amount of the compound of the invention administered
should be a therapeutically effective amount where the compound or
derivative is used for the treatment of a disease or condition and
a prophylactically effective amount where the compound or
derivative is used for the prevention of a disease or
condition.
[0116] The term "therapeutically effective amount" used herein
refers to the amount of compound needed to treat or ameliorate a
targeted disease or condition. The term "prophylactically effective
amount" used herein refers to the amount of compound needed to
prevent a targeted disease or condition. The exact dosage will
generally be dependent on the patient's status at the time of
administration. Factors that may be taken into consideration when
determining dosage include the severity of the disease state in the
patient, the general health of the patient, the age, weight,
gender, diet, time, frequency and route of administration, drug
combinations, reaction sensitivities and the patient's tolerance or
response to therapy. The precise amount can be determined by
routine experimentation, but may ultimately lie with the judgement
of the clinician. Generally, an effective dose will be from 0.01
mg/kg/day (mass of drug compared to mass of patient) to 1000
mg/kg/day, e.g. 1 mg/kg/day to 100 mg/kg/day. Compositions may be
administered individually to a patient or may be administered in
combination with other agents, drugs or hormones.
Administration & Formulation
General
[0117] For pharmaceutical use, the compounds of the invention may
be administered as a medicament by enteral or parenteral routes,
including intravenous, intramuscular, subcutaneous, transdermal,
airway (aerosol), oral, intranasal, rectal, vaginal and topical
(including buccal and sublingual) administration. The compounds of
formula (I) should be assessed for their biopharmaceutical
properties, such as solubility and solution stability (across pH),
permeability, etc., in order to select the most appropriate dosage
form and route of administration for treatment of the proposed
indication.
[0118] The compounds of the invention may be administered as
crystalline or amorphous products. The compounds of the invention
may be administered alone or in combination with one or more other
compounds of the invention or in combination with one or more other
drugs (or as any combination thereof). Generally, they will be
administered as a formulation in association with one or more
pharmaceutically acceptable excipients. The term "excipient"
includes any ingredient other than the compound(s) of the invention
which may impart either a functional (e.g. drug release rate
controlling) and/or a non-functional (e.g. processing aid or
diluent) characteristic to the formulations. The choice of
excipient will to a large extent depend on factors such as the
particular mode of administration, the effect of the excipient on
solubility and stability, and the nature of the dosage form.
[0119] Typical pharmaceutically acceptable excipients include:
[0120] diluents, e.g. lactose, dextrose, sucrose, mannitol,
sorbitol, cellulose and/or glycine; [0121] lubricants, e.g. silica,
talcum, stearic acid, its magnesium or calcium salt and/or
polyethyleneglycol; [0122] binders, e.g. magnesium aluminum
silicate, starch paste, gelatin, tragacanth, methylcellulose,
sodium carboxymethylcellulose and/or polyvinylpyrrolidone; [0123]
disintegrants, e.g. starches, agar, alginic acid or its sodium
salt, or effervescent mixtures; and/or [0124] absorbants,
colorants, flavors and/or sweeteners.
[0125] A thorough discussion of pharmaceutically acceptable
excipients is available in Gennaro, Remington: The Science and
Practice of Pharmacy 2000, 20th edition (ISBN: 0683306472).
Accordingly, in one embodiment, the present invention provides a
pharmaceutical composition comprising a compound of formula (I) and
a pharmaceutically acceptable excipient.
General Galenic Aspects
[0126] In another aspect, the present invention provides a
pharmaceutical composition comprising a compound of the present
invention, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier. The pharmaceutical composition
can be formulated for particular routes of administration such as
oral administration, parenteral administration, and rectal
administration, etc. In addition, the pharmaceutical compositions
of the present invention can be made up in a solid form (including
without limitation capsules, tablets, pills, granules, powders or
suppositories), or in a liquid form (including without limitation
solutions, suspensions or emulsions). The pharmaceutical
compositions can be subjected to conventional pharmaceutical
operations such as sterilization and/or can contain conventional
inert diluents, lubricating agents, or buffering agents, as well as
adjuvants, such as preservatives, stabilizers, wetting agents,
emulsifiers and buffers, etc.
[0127] Typically, the pharmaceutical compositions are tablets or
gelatin capsules comprising the active ingredient together with
a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol,
cellulose and/or glycine; b) lubricants, e.g., silica, talcum,
stearic acid, its magnesium or calcium salt and/or
polyethyleneglycol; for tablets also c) binders, e.g., magnesium
aluminum silicate, starch paste, gelatin, tragacanth,
methylcellulose, sodium carboxymethylcellulose and/or
polyvinylpyrrolidone; if desired d) disintegrants, e.g., starches,
agar, alginic acid or its sodium salt, or effervescent mixtures;
and/or e) absorbents, colorants, flavors and sweeteners.
[0128] Tablets may be either film coated or enteric coated
according to methods known in the art.
[0129] Suitable compositions for oral administration include an
effective amount of a compound of the invention in the form of
tablets, lozenges, aqueous or oily suspensions, dispersible powders
or granules, emulsion, hard or soft capsules, or syrups or elixirs.
Compositions intended for oral use are prepared according to any
method known in the art for the manufacture of pharmaceutical
compositions and such compositions can contain one or more agents
selected from the group consisting of sweetening agents, flavoring
agents, coloring agents and preserving agents in order to provide
pharmaceutically elegant and palatable preparations. Tablets may
contain the active ingredient in admixture with nontoxic
pharmaceutically acceptable excipients which are suitable for the
manufacture of tablets. These excipients are, for example, inert
diluents, such as calcium carbonate, sodium carbonate, lactose,
calcium phosphate or sodium phosphate; granulating and
disintegrating agents, for example, corn starch, or alginic acid;
binding agents, for example, starch, gelatin or acacia; and
lubricating agents, for example magnesium stearate, stearic acid or
talc. The tablets are uncoated or coated by known techniques to
delay disintegration and absorption in the gastrointestinal tract
and thereby provide a sustained action over a longer period. For
example, a time delay material such as glyceryl monostearate or
glyceryl distearate can be employed. Formulations for oral use can
be presented as hard gelatin capsules wherein the active ingredient
is mixed with an inert solid diluent, for example, calcium
carbonate, calcium phosphate or kaolin, or as soft gelatin capsules
wherein the active ingredient is mixed with water or an oil medium,
for example, peanut oil, liquid paraffin or olive oil.
[0130] Certain injectable compositions are aqueous isotonic
solutions or suspensions, and suppositories are advantageously
prepared from fatty emulsions or suspensions. Said compositions may
be sterilized and/or contain adjuvants, such as preserving,
stabilizing, wetting or emulsifying agents, solution promoters,
salts for regulating the osmotic pressure and/or buffers. In
addition, they may also contain other therapeutically valuable
substances. Said compositions are prepared according to
conventional mixing, granulating or coating methods, respectively,
and contain about 0.1-75%, or contain about 1-50%, of the active
ingredient.
[0131] Suitable compositions for transdermal application include an
effective amount of a compound of the invention with a suitable
carrier. Carriers suitable for transdermal delivery include
absorbable pharmacologically acceptable solvents to assist passage
through the skin of the host. For example, transdermal devices are
in the form of a bandage comprising a backing member, a reservoir
containing the compound optionally with carriers, optionally a rate
controlling barrier to deliver the compound of the skin of the host
at a controlled and predetermined rate over a prolonged period of
time, and means to secure the device to the skin.
[0132] Suitable compositions for topical application, e.g., to the
skin and eyes, include aqueous solutions, suspensions, ointments,
creams, gels or sprayable formulations, e.g., for delivery by
aerosol or the like. Such topical delivery systems will in
particular be appropriate for dermal application, e.g., for the
treatment of skin cancer, e.g., for prophylactic use in sun creams,
lotions, sprays and the like. They are thus particularly suited for
use in topical, including cosmetic, formulations well-known in the
art. Such may contain solubilizers, stabilizers, tonicity enhancing
agents, buffers and preservatives.
[0133] As used herein a topical application may also pertain to an
inhalation or to an intranasal application. They may be
conveniently delivered in the form of a dry powder (either alone,
as a mixture, for example a dry blend with lactose, or a mixed
component particle, for example with phospholipids) from a dry
powder inhaler or an aerosol spray presentation from a pressurised
container, pump, spray, atomizer or nebuliser, with or without the
use of a suitable propellant.
Dosage Forms
[0134] The pharmaceutical composition or combination of the present
invention can be in unit dosage of about 1-1000 mg of active
ingredient(s) for a subject of about 50-70 kg, or about 1-500 mg or
about 1-250 mg or about 1-150 mg or about 0.5-100 mg, or about 1-50
mg of active ingredients. The therapeutically effective dosage of a
compound, the pharmaceutical composition, or the combinations
thereof, is dependent on the species of the subject, the body
weight, age and individual condition, the disorder or disease or
the severity thereof being treated. A physician, clinician or
veterinarian of ordinary skill can readily determine the effective
amount of each of the active ingredients necessary to prevent,
treat or inhibit the progress of the disorder or disease.
[0135] The above-cited dosage properties are demonstrable in vitro
and in vivo tests using advantageously mammals, e.g., mice, rats,
dogs, monkeys or isolated organs, tissues and preparations thereof.
The compounds of the present invention can be applied in vitro in
the form of solutions, e.g., aqueous solutions, and in vivo either
enterally, parenterally, advantageously intravenously, e.g., as a
suspension or in aqueous solution. The dosage in vitro may range
between about 10.sup.-3 molar and 10.sup.-9 molar concentrations. A
therapeutically effective amount in vivo may range depending on the
route of administration, between about 0.1-500 mg/kg, or between
about 1-100 mg/kg.
[0136] The activity of a compound according to the present
invention can be assessed by the following in vitro & in vivo
methods.
Combination Therapy
[0137] The compound of formula (I) may be administered alone, or
may be administered in combination with another therapeutic agent
(i.e. a different agent to the compound of formula (I)).
Preferably, the compound of the invention and the other therapeutic
agent are administered in a therapeutically effective amount.
[0138] The compound of the present invention may be administered
either simultaneously with, or before or after, the other
therapeutic agent. The compound of the present invention may be
administered separately, by the same or different route of
administration, or together in the same pharmaceutical
composition.
[0139] In one embodiment, the invention provides a product
comprising a compound of formula (I) and another therapeutic agent
as a combined preparation for simultaneous, separate or sequential
use in therapy. In one embodiment, the therapy is the treatment of
a disease or condition mediated by DGAT1. Products provided as a
combined preparation include a composition comprising the compound
of formula (I) and the other therapeutic agent together in the same
pharmaceutical composition, or the compound of formula (I) and the
other therapeutic agent in separate form, e.g. in the form of a
kit.
[0140] In one embodiment, the invention provides a pharmaceutical
composition comprising a compound of formula (I) and another
therapeutic agent. Optionally, the pharmaceutical composition may
comprise a pharmaceutically acceptable excipient, as described
above in "Administration & Formulation".
[0141] In one embodiment, the invention provides a kit comprising
two or more separate pharmaceutical compositions, at least one of
which contains a compound of formula (I). In one embodiment, the
kit comprises means for separately retaining said compositions,
such as a container, divided bottle, or divided foil packet. An
example of such a kit is a blister pack, as typically used for the
packaging of tablets, capsules and the like.
[0142] The kit of the invention may be used for administering
different dosage forms, for example, oral and parenteral, for
administering the separate compositions at different dosage
intervals, or for titrating the separate compositions against one
another. To assist compliance, the kit of the invention typically
comprises directions for administration.
[0143] In the combination therapies of the invention, the compound
of the invention and the other therapeutic agent may be
manufactured and/or formulated by the same or different
manufacturers. Moreover, the compound of the invention and the
other therapeutic may be brought together into a combination
therapy: (i) prior to release of the combination product to
physicians (e.g. in the case of a kit comprising the compound of
the invention and the other therapeutic agent); (ii) by the
physician themselves (or under the guidance of the physician)
shortly before administration; (iii) in the patient themselves,
e.g. during sequential administration of the compound of the
invention and the other therapeutic agent.
[0144] Accordingly, the invention provides the use of a compound of
formula (I) in the manufacture of a medicament for treating a
disease or condition mediated by DGAT1, wherein the medicament is
prepared for administration with another therapeutic agent. The
invention also provides the use of a another therapeutic agent in
the manufacture of medicament for treating a disease or condition
mediated by DGAT1, wherein the medicament is prepared for
administration with a compound of formula (I).
[0145] The invention also provides a compound of formula (I) for
use in a method of treating a disease or condition mediated by
DGAT1, wherein the compound of formula (I) is prepared for
administration with another therapeutic agent. The invention also
provides another therapeutic agent for use in a method of treating
a disease or condition mediated by DGAT1, wherein the other
therapeutic agent is prepared for administration with a compound of
formula (I). The invention also provides a compound of formula (I)
for use in a method of treating a disease or condition mediated by
DGAT1, wherein the compound of formula (I) is administered with
another therapeutic agent. The invention also provides another
therapeutic agent for use in a method of treating a disease or
condition mediated by DGAT1, wherein the other therapeutic agent is
administered with a compound of formula (I).
[0146] The invention also provides the use of a compound of formula
(I) in the manufacture of a medicament for treating a disease or
condition mediated by DGAT1, wherein the patient has previously
(e.g. within 24 hours) been treated with another therapeutic agent.
The invention also provides the use of another therapeutic agent in
the manufacture of a medicament for treating a disease or condition
mediated by DGAT1, wherein the patient has previously (e.g. within
24 hours) been treated with a compound of formula (I).
[0147] In one embodiment, the other therapeutic agent is selected
from: [0148] antidiabetic agents, such as insulin, insulin
derivatives and mimetics; insulin secretagogues such as the
sulfonylureas, e.g. Glipizide, glyburide and Amaryl; insulinotropic
sulfonylurea receptor ligands such as meglitinides, e.g.
nateglinide and repaglinide; protein tyrosine phosphatase-1B
(PTP-1B) inhibitors such as PTP-112; Cholesteryl ester transfer
protein (CETP) inhibitors such as torcetrapib, GSK3 (glycogen
synthase kinase-3) inhibitors such as SB-517955, SB-4195052,
SB-216763, NN-57-05441 and NN-57-05445; RXR ligands such as GW-0791
and AGN-194204; sodium-dependent glucose cotransporter inhibitors
such as T-1095; glycogen phosphorylase A inhibitors such as BAY
R3401; biguanides such as metformin; alpha-glucosidase inhibitors
such as acarbose; GLP-1 (glucagon like peptide-1), GLP-1 analogs
such as Exendin-4 and GLP-1 mimetics; and DPPIV (dipeptidyl
peptidase IV) inhibitors such as vildagliptin; [0149] hypolipidemic
agents such as 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA)
reductase inhibitors, e.g. lovastatin, pitavastatin, simvastatin,
pravastatin, cerivastatin, mevastatin, velostatin, fluvastatin,
dalvastatin, atorvastatin, rosuvastatin and rivastatin; squalene
synthase inhibitors; FXR (farnesoid X receptor) and LXR (liver X
receptor) ligands; cholestyramine; fibrates; nicotinic acid and
aspirin; [0150] anti-obesity agents such as orlistat or rimonabant;
[0151] anti-hypertensive agents, e.g. loop diuretics such as
ethacrynic acid, furosemide and torsemide; angiotensin converting
enzyme (ACE) inhibitors such as benazepril, captopril, enalapril,
fosinopril, lisinopril, moexipril, perinodopril, quinapril,
ramipril and trandolapril; inhibitors of the Na--K-ATPase membrane
pump such as digoxin; neutralendopeptidase (NEP) inhibitors;
ACE/NEP inhibitors such as omapatrilat, sampatrilat and fasidotril;
angiotensin II antagonists such as candesartan, eprosartan,
irbesartan, losartan, telmisartan and valsartan, in particular
valsartan; renin inhibitors such as ditekiren, zankiren,
terlakiren, aliskiren, RO 66-1132 and RO-66-1168; .beta.-adrenergic
receptor blockers such as acebutolol, atenolol, betaxolol,
bisoprolol, metoprolol, nadolol, propranolol, sotalol and timolol;
inotropic agents such as digoxin, dobutamine and milrinone; calcium
channel blockers such as amlodipine, bepridil, diltiazem,
felodipine, nicardipine, nimodipine, nifedipine, nisoldipine and
verapamil; aldosterone receptor antagonists; and aldosterone
synthase inhibitors; [0152] agonists of peroxisome
proliferator-activator receptors, such as fenofibrate,
pioglitazone, rosiglitazone, tesaglitazar, BMS-298585, L-796449,
the compounds specifically described in the patent application WO
2004/103995 i.e. compounds of examples 1 to 35 or compounds
specifically listed in claim 21, or the compounds specifically
described in the patent application WO 03/043985 i.e. compounds of
examples 1 to 7 or compounds specifically listed in claim 19 and
especially
(R)-1-{4-[5-methyl-2-(4-trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benze-
nesulfonyl}-2,3-dihydro-1H-indole-2-carboxylic or a salt thereof;
and [0153] the specific anti-diabetic compounds described in Expert
Opin Investig Drugs 2003, 12(4): 623-633, FIGS. 1 to 7.
General Methods of Preparation
[0154] Specific methods for the preparation of the compounds of the
invention are disclosed in detail below in the Examples.
[0155] In general, compounds of formula (I) may be prepared by the
reaction schemes described below.
[0156] Compound of the invention in which ring A is a oxadiazole
can be prepared by reacting the aniline (1) with
1,1'-thiocarbonyldipyridin-2(1H)-one (2). The isothiocyanate (3) is
then reacted with carbohydrazides (4) to form the
hydrazinecarbothioamide (5). Cyclization with
3-(ethyliminomethyleneamino)-N,N-dimethyl-propan-1-amine (EDC) to
form a compound of the invention represented by formula I.
Compounds represented by formula IA in which ring A is a oxadiazole
and ring B is a phenyl can be prepared using the reactions of
Scheme I but also in the alternative method shown in Scheme II and
III.
[0157] Compounds represented by formula IA in which ring A is
oxadiazole and B is phenyl can be prepared from the Suzuki coupling
of the pinacol boronate ester (6) and the corresponding biphenyl
bromide (7), as indicated in Scheme II. Alternatively, compounds
represented by formula IA in which ring A is oxadizole and B is
pyridine can be prepared by reacting the oxadiazole (8) with the
corresponding pinacol boronate ester (9), as indicated in Scheme
III. Compounds represented by formula IIA in which ring A is a
thiadiazoles can be prepared using the reactions in scheme 1.
Cyclization of the hydrazinecarbothioamide with sulphuric acid in
the presence of ethanol afford the compound of the invention
represented by formula IIA.
##STR00009##
##STR00010##
##STR00011##
[0158] In general, compounds of formula (IVA) and (VA) may be
prepared by the reaction schemes below.
##STR00012## ##STR00013##
[0159] 4-phenyl-2-oxabicyclo[2.2.2]octane can be prepared by
reacting the ketone (12) with methyl acrylate (13). Subsequent ring
closure followed by decarboxylation afforded the cyclohexanone
compound (14), which is protected to form the acetal (15).
Reduction, followed by deprotection of the acetal formed the
primary alcohol (17). Reaction with trimethyl phosphonacetate,
followed by base promoted intra-molecular cyclization allowed the
formation of a compound of the invention represented by formula
IVA.
[0160] 1-phenyl-2-oxabicyclo[2.2.2]octane can be prepared by
reacting the ketone (20) with Comin's Reagent in the presence of
base to afford intermediate (21). Suzuki coupling with the boronic
acid (22) followed by reduction and cyclization afforded the
primary alcohol (24). Oxidation, followed by homologation afforded
the enol ether (25). Oxidation and subsequent reduction allowed the
formation of a compound of the invention represented by formula
VA.
##STR00014## ##STR00015##
[0161] In general, compounds of formula (XIA) may be prepared by
the reaction scheme below.
[0162] 7-oxabicyclo[2.2.1]heptan-1-yl compounds can be prepared by
reacting the ketone (47) with the in-situ generated Grignard
reagent (48), affording the tertiary alcohol (49). Deprotection,
followed by Wittig generated the ester (50). Finally, a base
promoted intra-molecular cyclization allowed the formation of a
compound of the invention represented by formula XIA.
##STR00016##
[0163] In general, compounds of formula (XIIA) may be prepared by
the reaction scheme below.
[0164] 7-oxabicyclo[2.2.1]heptan-1-yl propanoic acid compounds can
be prepared by reacting the ketone (47) with phosphonium-ylide via
Wittig reaction, followed by deprotection to afford the alkene
(51). Reaction with the in-situ generated Grignard reagent (48)
afforded the tertiary alcohol (52). Iodine promoted cyclization
generated the bicyclo ring intermediate (53), which can be
converted to the bis-methyl ester (55) via alkylation with dimethyl
malonate (54). Final decarboxylation step allowed the formation of
a compound of the invention represented by formula XIIA.
##STR00017##
Chemical Groups
[0165] As used herein, the term "halogen" (or halo) refers to
fluorine, bromine, chlorine or iodine, in particular fluorine,
chlorine. Halogen-substituted groups and moieties, such as alkyl
substituted by halogen (haloalkyl) can be mono-, poly- or
per-halogenated.
[0166] As used herein, the term "hetero atoms" refers to nitrogen
(N), oxygen (O) or sulphur (S) atoms, in particular nitrogen or
oxygen.
[0167] As used herein, the term "alkyl" refers to a fully saturated
branched or unbranched hydrocarbon moiety having up to 20 carbon
atoms. Unless otherwise provided, alkyl refers to hydrocarbon
moieties having 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 7
carbon atoms, or 1 to 4 carbon atoms. Representative examples of
alkyl include, but are not limited to, methyl, ethyl, n-propyl,
iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl,
isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl,
2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl and the
like. A substituted alkyl is an alkyl group containing one or more,
such as one, two or three substituents selected from halogen,
hydroxy or alkoxy groups.
[0168] As used herein, the term "alkylene" refers to divalent alkyl
group as defined herein above having 1 to 20 carbon atoms. It
comprises 1 to 20 carbon atoms, Unless otherwise provided, alkylene
refers to moieties having 1 to 16 carbon atoms, 1 to 10 carbon
atoms, 1 to 7 carbon atoms, or 1 to 4 carbon atoms. Representative
examples of alkylene include, but are not limited to, methylene,
ethylene, n-propylene, iso-propylene, n-butylene, sec-butylene,
iso-butylene, tert-butylene, n-pentylene, isopentylene,
neopentylene, n-hexylene, 3-methylhexylene, 2,2-dimethylpentylene,
2,3-dimethylpentylene, n-heptylene, n-octylene, n-nonylene,
n-decylene and the like.
[0169] A substituted alkylene is an alkylene group containing one
or more, such as one, two or three substituents selected from
halogen, hydroxy or alkoxy groups.
[0170] As used herein, the term "haloalkyl" refers to an alkyl as
defined herein, which is substituted by one or more halo groups as
defined herein. The haloalkyl can be monohaloalkyl, dihaloalkyl or
polyhaloalkyl including perhaloalkyl. A monohaloalkyl can have one
iodo, bromo, chloro or fluoro within the alkyl group. Dihaloalky
and polyhaloalkyl groups can have two or more of the same halo
atoms or a combination of different halo groups within the alkyl.
Typically the polyhaloalkyl contains up to 12, or 10, or 8, or 6,
or 4, or 3, or 2 halo groups. Non-limiting examples of haloalkyl
include fluoromethyl, difluoromethyl, trifluoromethyl,
chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl,
heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl,
difluoroethyl, difluoropropyl, dichloroethyl and dichloropropyl. A
perhalo-alkyl refers to an alkyl having all hydrogen atoms replaced
with halo atoms.
[0171] As used herein, the term "alkoxy" refers to alkyl-O--,
wherein alkyl is defined herein above. Representative examples of
alkoxy include, but are not limited to, methoxy, ethoxy, propoxy,
2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy,
cyclopropyloxy-, cyclohexyloxy- and the like. Typically, alkoxy
groups have 1-16, 1-10, 1-7, more preferably 1-4 carbon atoms.
[0172] A substituted alkoxy is an alkoxy group containing one or
more, such as one, two or three substituents selected from halogen,
hydroxy or alkoxy groups.
[0173] Similarly, each alkyl part of other groups like
"alkylaminocrabonyl", "alkoxyalkyl", "alkoxycarbonyl",
"alkoxy-carbonylalkyl", "alkylsulfonyl", "alkylsulfoxyl",
"alkylamino", "haloalkyl" shall have the same meaning as described
in the above-mentioned definition of "alkyl".
[0174] As used herein, the term "cycloalkyl" refers to saturated or
unsaturated monocyclic, bicyclic, tricyclic or spirocyclic
hydrocarbon groups of 3-12 carbon atoms. Unless otherwise provided,
cycloalkyl refers to cyclic hydrocarbon groups having between 3 and
9 ring carbon atoms or between 3 and 7 ring carbon atoms.
[0175] A substituted cycloalkyl is a cycloalkyl group substituted
by one, or two, or three, or more substituents independently
selected from the group consisting of hydroxyl, thiol, cyano,
nitro, oxo, alkylimino, C.sub.1-C.sub.4-alkyl,
C.sub.1-C.sub.4-alkenyl, C.sub.1-C.sub.4-alkynyl,
C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkyl,
C.sub.1-C.sub.4-alkenyloxy, C.sub.1-C.sub.4-alkynyloxy, halogen,
C.sub.1-C.sub.4alkylcarbonyl, carboxy,
C.sub.1-C.sub.4alkoxycarbonyl, amino, C.sub.1-C.sub.4-alkylamino,
di-C.sub.1-C.sub.4-alkylamino, C.sub.1-C.sub.4-alkylaminocarbonyl,
di-C.sub.1-C.sub.4-alkylaminocarbonyl,
C.sub.1-C.sub.4-alkylcarbonylamino,
C.sub.1-C.sub.4-alkylcarbonyl(C.sub.1-C.sub.4-alkyl)amino,
sulfonyl, sulfamoyl, alkylsulfamoyl,
C.sub.1-C.sub.4alkylaminosulfonyl where each of the afore-mentioned
hydrocarbon groups (e.g., alkyl, alkenyl, alkynyl, alkoxy residues)
may be further substituted by one or more residues independently
selected at each occurrence from halogen, hydroxyl or
C.sub.1-C.sub.4-alkoxy groups. Exemplary monocyclic hydrocarbon
groups include, but are not limited to, cyclopropyl, cyclobutyl,
cyclopentyl, cyclopentenyl, cyclohexyl and cyclohexenyl and the
like. Exemplary bicyclic hydrocarbon groups include bornyl, indyl,
hexahydroindyl, tetrahydronaphthyl, decahydronaphthyl,
bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl,
6,6-dimethylbicyclo[3.1.1]heptyl,
2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl and the
like. Exemplary tricyclic hydrocarbon groups include adamantyl and
the like.
[0176] Similarly, each cycloalkyl part of other groups like
"cycloalkyloxy", "cycloalkoxyalkyl", "cycloalkoxycarbonyl",
"cycloalkoxy-carbonylalkyl", "cycloalkylsulfonyl", "halocycloalkyl"
shall have the same meaning as described in the above-mentioned
definition of "alkyl".
[0177] As used herein, the term "aryl" refers to an aromatic
hydrocarbon group having 6-20 carbon atoms in the ring portion.
Typically, aryl is monocyclic, bicyclic or tricyclic aryl having
6-20 carbon atoms. Furthermore, the term "aryl" as used herein,
refers to an aromatic substituent which can be a single aromatic
ring, or multiple aromatic rings that are fused together.
Non-limiting examples include phenyl, naphthyl or
tetrahydronaphthyl.
[0178] A substituted aryl is an aryl group substituted by 1-5 (such
as one, or two, or three) substituents independently selected from
the group consisting of hydroxyl, thiol, cyano, nitro,
C.sub.1-C.sub.4-alkyl, C.sub.1-C.sub.4-alkenyl,
C.sub.1-C.sub.4-alkynyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkyl, C.sub.1-C.sub.4-alkenyloxy,
C.sub.1-C.sub.4-alkynyloxy, halogen, C.sub.1-C.sub.4-alkylcarbonyl,
carboxy, C.sub.1-C.sub.4-alkoxycarbonyl, amino,
C.sub.1-C.sub.4-alkylamino, di-C.sub.1-C.sub.4-alkylamino,
C.sub.1-C.sub.4alkylaminocarbonyl,
di-C.sub.1-C.sub.4alkylaminocarbonyl,
C.sub.1-C.sub.4-alkylcarbonylamino,
C.sub.1-C.sub.4-alkylcarbonyl(C.sub.1-C.sub.4-alkyl)amino,
sulfonyl, sulfamoyl, alkylsulfamoyl,
C.sub.1-C.sub.4-alkylaminosulfonyl where each of the
afore-mentioned hydrocarbon groups (e.g., alkyl, alkenyl, alkynyl,
alkoxy residues) may be further substituted by one or more residues
independently selected at each occurrence from halogen, hydroxyl or
C.sub.1-C.sub.4-alkoxy groups.
[0179] Similarly, each aryl part of other groups like "aryloxy",
"aryloxyalkyl", "aryloxycarbonyl", "aryloxy-carbonylalkyl" shall
have the same meaning as described in the above-mentioned
definition of "aryl".
[0180] As used herein, the term "heterocyclyl" refers to a
heterocyclic radical that saturated or partially saturated and is
preferably a monocyclic or a polycyclic ring (in case of a
polycyclic ring particularly a bicyclic, tricyclic or spirocyclic
ring); and has 3 to 24, more preferably 4 to 16, most preferably 5
to 10 and most preferably 5 or 6 ring atoms; wherein one or more,
preferably one to four, especially one or two ring atoms are a
heteroatom (the remaining ring atoms therefore being carbon). The
bonding ring (i.e. the ring connecting to the molecule) preferably
has 4 to 12, especially 5 to 7 ring atoms. The term heterocyclyl
excludes heteroaryl. The heterocyclic group can be attached at a
heteroatom or a carbon atom. The heterocyclyl can include fused or
bridged rings as well as spirocyclic rings. Examples of
heterocycles include tetrahydrofuran (THF), dihydrofuran,
1,4-dioxane, morpholine, 1,4-dithiane, piperazine, piperidine,
1,3-dioxolane, imidazolidine, imidazoline, pyrroline, pyrrolidine,
tetrahydropyran, dihydropyran, oxathiolane, dithiolane,
1,3-dioxane, 1,3-dithiane, oxathiane, thiomorpholine, and the
like
[0181] A substituted heterocyclyl is a heterocyclyl group
independently substituted by 1-5 (such as one, or two, or three)
substituents selected from hydroxyl, thiol, cyano, nitro, oxo,
alkylimino, C.sub.1-C.sub.4-alkyl, C.sub.1-C.sub.4-alkenyl,
C.sub.1-C.sub.4-alkynyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkyl, C.sub.1-C.sub.4-alkenyloxy,
C.sub.1-C.sub.4-alkynyloxy, halogen, C.sub.1-C.sub.4alkylcarbonyl,
carboxy, C.sub.1-C.sub.4-alkoxycarbonyl, amino,
C.sub.1-C.sub.4-alkylamino, di-C.sub.1-C.sub.4alkylamino,
C.sub.1-C.sub.4alkylaminocarbonyl,
di-C.sub.1-C.sub.4alkylaminocarbonyl,
C.sub.1-C.sub.4-alkylcarbonylamino,
C.sub.1-C.sub.4alkylcarbonyl(C.sub.1-C.sub.4-alkyl)amino, sulfonyl,
sulfamoyl, alkylsulfamoyl, C.sub.1-C.sub.4alkylaminosulfonyl where
each of the afore-mentioned hydrocarbon groups (e.g., alkyl,
alkenyl, alkynyl, alkoxy residues) may be further substituted by
one or more residues independently selected at each occurrence from
halogen, hydroxyl or C.sub.1-C.sub.4-alkoxy groups.
[0182] Similarly, each heterocyclyl part of other groups like
"heterocyclyloxy", "heterocyclyloxyalkyl",
"heterocyclyloxycarbonyl" shall have the same meaning as described
in the above-mentioned definition of "heterocyclyl".
[0183] As used herein, the term "heteroaryl" refers to a 5-14
membered monocyclic- or bicyclic- or tricyclic-aromatic ring
system, having 1 to 8 heteroatoms. Typically, the heteroaryl is a
5-10 membered ring system (e.g., 5-7 membered monocycle or an 8-10
membered bicycle) or a 5-7 membered ring system. Typical heteroaryl
groups include 2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl,
2-, 4-, or 5-imidazolyl, 3-, 4-, or 5-pyrazolyl, 2-, 4-, or
5-thiazolyl, 3-, 4-, or 5-isothiazolyl, 2-, 4-, or 5-oxazolyl, 3-,
4-, or 5-isoxazolyl, 3- or 5-1,2,4-triazolyl, 4- or
5-1,2,3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridyl, 3- or
4-pyridazinyl, 3-, 4-, or 5-pyrazinyl, 2-pyrazinyl, and 2-, 4-, or
5-pyrimidinyl.
[0184] The term "heteroaryl" also refers to a group in which a
heteroaromatic ring is fused to one or more aryl, cycloaliphatic,
or heterocyclyl rings, where the radical or point of attachment is
on the heteroaromatic ring. Nonlimiting examples include 1-, 2-,
3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or
7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-,
or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-,
6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-, or
8-quinoliyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinoliyl, 1-, 4-, 5-,
6-, 7-, or 8-phthalazinyl, 2-, 3-, 4-, 5-, or 6-naphthyridinyl, 2-,
3-, 5-, 6-, 7-, or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7-, or
8-cinnolinyl, 2-, 4-, 6-, or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-,
7-, or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or
8-carbzaolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-carbolinyl, 1-, 2-,
3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl, 1-, 2-, 3-, 4-, 5-,
6-, 7-, 8-, or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or
9-perimidinyl, 2-, 3-, 4-, 5-, 6-, 8-, 9-, or 10-phenathrolinyl,
1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-,
7-, 8-, 9-, or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-,
or 10-phenoxazinyl, 2-, 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-,
8-, 9-, or 10-benzisoqinolinyl, 2-, 3-, 4-, or
thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or
11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6-, or
7-2H-furo[3,2-b]-pyranyl, 2-, 3-, 4-, 5-, 7-, or
8-5H-pyrido[2,3-d]-o-oxazinyl, 1-, 3-, or
5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or
54H-imidazo[4,5-d]thiazolyl, 3-, 5-, or
8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5-, or
6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8-, or
9-furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10, or
11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6-, or
7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-,
6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-,
4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or
9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-,
5-, 6-, 7-, 8-, 9-, 10-, or 11-1H-pyrrolo[1,2-b][2]benzazapinyl.
Typical fused heteroary groups include, but are not limited to 2-,
3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or
8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-,
6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-,
4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or
7-benzothiazolyl.
[0185] A substituted heteroaryl is a heteroaryl group containing
one or more substituents selected from hydroxyl, thiol, cyano,
nitro, C.sub.1-C.sub.4-alkyl, C.sub.1-C.sub.4-alkenyl,
C.sub.1-C.sub.4-alkynyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkyl, C.sub.1-C.sub.4alkenyloxy,
C.sub.1-C.sub.4alkynyloxy, halogen, C.sub.1-C.sub.4-alkylcarbonyl,
carboxy, C.sub.1-C.sub.4-alkoxycarbonyl, amino,
C.sub.1-C.sub.4-alkylamino, di-C.sub.1-C.sub.4-alkylamino,
C.sub.1-C.sub.4-alkylaminocarbonyl,
di-C.sub.1-C.sub.4-alkylaminocarbonyl,
C.sub.1-C.sub.4-alkylcarbonylamino,
C.sub.1-C.sub.4-alkylcarbonyl(C.sub.1-C.sub.4-alkyl)amino,
sulfonyl, sulfamoyl, alkylsulfamoyl,
C.sub.1-C.sub.4-alkylaminosulfonyl where each of the
afore-mentioned hydrocarbon groups (e.g., alkyl, alkenyl, alkynyl,
alkoxy residues) may be further substituted by one or more residues
independently selected at each occurrence from halogen, hydroxyl or
C.sub.1-C.sub.4-alkoxy groups.
[0186] Similarly, each heteroaryl part of other groups like
"heteroaryloxy", "heteroaryloxyalkyl", "heteroaryloxycarbonyl"
shall have the same meaning as described in the above-mentioned
definition of "heteroaryl".
General
[0187] The term "comprising" encompasses "including" as well as
"consisting" e.g. a composition "comprising" X may consist
exclusively of X or may include something additional e.g. X+Y.
[0188] The word "substantially" does not exclude "completely" e.g.
a composition which is "substantially free" from Y may be
completely free from Y. Where necessary, the word "substantially"
may be omitted from the definition of the invention.
[0189] The term "about" in relation to a numerical value x means,
for example, x.+-.10%.
MODES FOR CARRYING OUT THE INVENTION
[0190] The following Examples are intended to illustrate the
invention and are not to be construed as being limitations thereon.
If not mentioned otherwise, all evaporations are performed under
reduced pressure, between about 50 mmHg and 100 mmHg. The structure
of final products, intermediates and starting materials is
confirmed by standard analytical methods, e.g., microanalysis,
melting point (m.p.) and spectroscopic characteristics, e.g. MS, IR
and NMR. Abbreviations used are those conventional in the art.
[0191] The conditions for determining the mass and the retention
times were as follows:
Condition E (LCMS: RXNMON_Neutral)
[0192] LC-MS method with Broad range (5-95%) gradient with neutral
mobile phase (5 mM NH4+HCOO-). Electrospray mass spectra (+) and
(-), DAD-UV chromatogram 210-400 nm, Gradient: 5-95% MeCN in 2 min
(2 mL/min), 2 .mu.L injection. Column: Inertsil C.sub.8-3,
3.0.times.433 mm.times.3.0 .mu.m, 40 deg C.
Condition Z (HR/MS, Pre OAA)
[0193] 1.0 mL/min flow rate. 5% to 95% Acetonitrile (with 0.05%
formic acid) gradient in 9.50 min, Aqueous phase modified with 0.1%
formic acid. Column: Inertsil ODS-4 C18, 3 um, 3.0.times.100 mm.
LCUV/ESI-MS data was recorded on an Agilent 6220 with resolution of
11000 (FWHM).
Condition L (QT2, OAA HR/MS)
[0194] 1.0 mL/min flow rate with the gradient from 2% to 98% ACN in
4.40 min, 3.75 mM Ammonium Acetate and 0.0005% Formic Acid used as
the modifier additive in the Aqueous Phase. 0.04% of Formic Acid
used as the modifier in the Organic Phase. Acquity UPLC CSH Ca18
2.1.times.50 mm 1.7 um column at 50 deg C., LCUV/ESI-MS data was
recorded on an Acquity G2 Xevo QTof with resolution of >20000
(FWHM).
Condition M (SQ2, Purity-NpH)
[0195] 1.0 mL/min flow rate with the gradient from 2% to 98% ACN in
4.40 min, 3.75 mM Ammonium Acetate and 2% acetonitrile used as the
modifier additive in the Aqueous Phase. No additive was used as the
modifier in the Organic Phase. Acquity UPLC CSH C18 2.1.times.50 mm
1.7 um column at 50 deg C.
Condition R (LCMS: RXNMON_Neutral, ZQ1)
[0196] LC-MS method with Broad range (5-95%) gradient with neutral
mobile phase (5 mM NH4+HCOO-). Electrospray mass spectra (+) and
(-), DAD-UV chromatogram 210-400 nm, Gradient: 5-95% MeCN in 2 min
(2 mL/min), 2 .mu.L injection. Column: X-bridge C18, 3.0
cm.times.30 mm.times.3.5 .mu.m, 40.degree. C.
Condition W (SQ4, RXNMON-Acidic)
[0197] 1.0 mL/min flow rate with the gradient from 2% to 98% ACN in
1.70 min, 3.75 mM Ammonium Acetate and 0.05% Formic Acid used as
the modifier additive in the Aqueous Phase. 0.04% of Formic Acid
used as the modifier in the Organic Phase. Acquity UPLC BEH C18
2.1.times.50 mm 1.7 um column at 50 deg C.
TABLE-US-00001 Ex. # MW Structure IUPAC Name 1 462.54 ##STR00018##
2-(4-(4-(5-(5-tert-butyl-1,3,4- oxadiazol-2-ylamino)pyridin-
2-yl)phenyl)-2- oxabicyclo[2.2.2]octan-1- yl)acetic acid 2 460.52
##STR00019## 2-(4-(4-(5-((5-cyclobutyl- 1,3,4-oxadiazol-2-
yl)amino)pyridin-2- yl)phenyl)-2- oxabicyclo[2.2.2]octan-1-
yl)acetic acid 3 476.59 ##STR00020## 2-(4-(4-(5-((5-cyclobutyl-
1,3,4-thiadiazol-2- yl)amino)pyridin-2- yl)phenyl)-2-
oxabicyclo[2.2.2]octan-1- yl)acetic acid 4 477.58 ##STR00021##
2-(4-(4-(2-(5-cyclobutyl- 1,3,4-thiadiazol-2- ylamino)pyrimidin-5-
yl)phenyl)-2- oxabicyclo[2.2.2]octan-1- yl)acetic acid 5 476.59
##STR00022## 2-(4-(4-(6-((5-cyclobutyl- 1,3,4-thiadiazol-2-
yl)amino)pyridin-3- yl)phenyl)-2- oxabicyclo[2.2.2]octan-1-
yl)acetic acid 6 475.60 ##STR00023## 2-(4-(4'-(5-cyclobutyl-1,3,4-
thiadiazol-2- ylamino)biphenyl-4-yl)-2- oxabicyclo[2.2.2]octan-1-
yl)acetic acid 7 459.54 ##STR00024## 2-(4-(4'-(5-cyclobutyl-1,3,4-
oxadiazol-2- ylamino)biphenyl-4-yl)-2- oxabicyclo[2.2.2]octan-1-
yl)acetic acid 8 461.55 ##STR00025## 2-(4-(4'-(5-tert-butyl-1,3,4-
oxadiazol-2- ylamino)biphenyl-4-yl)-2- oxabicyclo[2.2.2]octan-1-
yl)acetic acid 9 445.51 ##STR00026## 2-(4-(4'-((5-cyclopropyl-
1,3,4-oxadiazol-2-yl)amino)- [1,1'-biphenyl]-4-yl)-2-
oxabicyclo[2.2.2]octan-1- yl)acetic acid 10 514.49 ##STR00027##
2-(4-(4-(5-(2-methyl-5- (trifluoromethyl)oxazole-4-
carboxamido)pyridin-2- yl)phenyl)-2- oxabicyclo[2.2.2]octan-1-
yl)acetic acid 11 474.55 ##STR00028## 2-(4-(4'-(2-ethyl-N,4-
dimethyloxazole-5- carboxamido)-[1,1'-biphenyl]- 4-yl)-2-
oxabicyclo[2.2.2]octan-1- yl)acetic acid 12 515.48 ##STR00029##
2-(4-(4-(5-(2-methyl-5- (trifluoromethyl)oxazole-4-
carboxamido)pyridin-2- yl)phenyl)-2- oxabicyclo[2.2.2]octan-1-
yl)acetic acid 13 475.54 ##STR00030## 2-(4-(4-(5-(2-ethyl-4-
methyloxazole-5- carboxamido)pyridin-2- yl)phenyl)-2-
oxabicyclo[2.2.2]octan-1- yl)acetic acid 14 461.55 ##STR00031##
2-(4-(4-(5-((5-(tert- butyl)oxazol-2- yl)amino)pyridin-2-
yl)phenyl)-2- oxabicyclo[2.2.2]octan-1- yl)acetic acid 15 460.56
##STR00032## 2-(4-(4'-((5-(tert- butyl)oxazol-2-yl)amino)-
[1,1'-biphenyl]-4-yl)-2- oxabicyclo[2.2.2]octan-1- yl)acetic acid
16 461.55 ##STR00033## 2-(4-(4'-((5-isobutyl-1,3,4-
oxadiazol-2-yl)amino)-[1,1'- biphenyl]-4-yl)-2-
oxabicyclo[2.2.2]octan-1- yl)acetic acid 17 475.58 ##STR00034##
2-(4-(4'-((5-neopentyl-1,3,4- oxadiazol-2-yl)amino)-[1,1'-
biphenyl]-4-yl)-2- oxabicyclo[2.2.2]octan-1- yl)acetic acid 18
474.55 ##STR00035## 2-(1-(4'-(2-ethyl-4- methyloxazole-5-
carboxamido)-[1,1'-biphenyl]- 4-yl)-2- oxabicyclo[2.2.2]octan-4-
yl)acetic acid 19 459.54 ##STR00036##
2-(1-(4'-((5-cyclobutyl-1,3,4- oxadiazol-2-yl)amino)-[1,1'-
biphenyl]-4-yl)-2- oxabicyclo[2.2.2]octan-4- yl)acetic acid 20
461.55 ##STR00037## 2-(1-(4'-((5-(tert-butyl)-1,3,4-
oxadiazol-2-yl)amino)-[1,1'- biphenyl]-4-yl)-2-
oxabicyclo[2.2.2]octan-4- yl)acetic acid 21 460.52 ##STR00038##
2-(1-(4-(5-((5-cyclobutyl- 1,3,4-oxadiazol-2- yl)amino)pyridin-2-
yl)phenyl)-2- oxabicyclo[2.2.2]octan-4- yl)acetic acid 22 447.53
##STR00039## 2-(4-(4'-((5-(tert-butyl)-1,3,4-
oxadiazol-2-yl)amino)-[1,1'- biphenyl]-4-yl)-7-
oxabicyclo[2.2.1]heptan-1- yl)acetic acid 23 461.55 ##STR00040##
3-(4-(4'-((5-(tert-butyl)-1,3,4- oxadiazol-2-yl)amino)-[1,1'-
biphenyl]-4-yl)-7- oxabicyclo[2.2.1]heptan-1- yl)propanoic acid
Example 1
2-(4-(4-(5-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)pyridin-2-yl)phenyl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
1-(4-bromophenyl)-4-oxocyclohexanecarboxylate
##STR00041##
[0199] The title compound was prepared analogous to the literature
procedure (J. Org. Chem., 72, 7455, 2007) starting from methyl
2-(4-bromophenyl)acetate (5.78 g, 25.2 mmol) and methyl acrylate
(4.78 g, 55.5 m mol). Column purification afforded the title
compound as clear oil (4.64 g, 59% yield). LC/MS, ESI-MS(+): 311.1,
RT: 1.26 (Condition E).
Step 2. Synthesis of methyl
8-(4-bromophenyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate
##STR00042##
[0201] To a round bottom flask containing methyl
1-(4-bromophenyl)-4-oxocyclohexanecarboxylate (4.64 g, 14.91 mmol)
was added toluene (33 ml), ethylene glycol (8.32 ml, 149 mmol) and
p-toluenesulfonic acid (85 mg, 0.45 mmol) at room temperature. The
mixture was heated to 80 C for 2 hours. The content was then cooled
to room temperature and quenched with a saturated solution of
sodium bicarbonate. The organic portion was washed with saturated
sodium bicarbonate and the aqueous portion extracted with MTBE
twice. The combined organic portion was dried over sodium sulfate
and concentrated to afford a crude oil. Column purification
afforded the title compound as viscous oil (5.30 g, quantitative
yield). LC/MS, ESI-MS(+): 357.1, RT: 1.38 (Condition E).
Step 3. Synthesis of
(8-(4-bromophenyl)-1,4-dioxaspiro[4.5]decan-8-yl)methanol
##STR00043##
[0203] To a round bottom flask containing methyl
8-(4-bromophenyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate (5.30 g,
14.92 mmol) was added anhydrous dichloromethane (60 ml) at room
temperature. The content was then cool to -78.degree. C. under a
dry ice-acetone bath and DIBAL-H (31.1 ml, 37.30 mmol) in toluene
was added dropwise slowly. When the addition was completed, the
mixture was stirred at -78.degree. C. for 30 minutes or until the
consumption of the starting material as indicated by thin layer
chromatography (TLC). The reaction was then quenched by the slow
addition of a pH 8 buffered solution (prepared by mixing 0.53 mL
ammonium hydroxide solution and 8.8 mL of saturated ammonium
chloride solution) at -78.degree. C. The mixture was then allowed
to warm back up to room temperature and stirred for a further 45
minutes. Solid magnesium sulfate (8 g) was added and the mixture
stirred at room temperature for another 30 minutes to afford free
flowing slurry. The content was filtered and concentrated to afford
the title compound as clear oil, which was used in the next step
without purification (4.88 g). LC/MS, ESI-MS: non-ionizable, RT:
1.05 (Condition E).
Step 4. Synthesis of
4-(4-bromophenyl)-4-(hydroxymethyl)cyclohexanone
##STR00044##
[0205] To a round bottom flask containing
(8-(4-bromophenyl)-1,4-dioxaspiro[4.5]decan-8-yl)methanol (4.88 g,
14.91 mmol) was added acetone (34 ml) and water (17 ml) at room
temperature. p-toluenesulfonic acid (57 mg, 0.30 mmol) was then
added and the mixture heated to 75.degree. C. for 1 hour. The
content was cooled to room temperature and concentrated under
reduced pressure to remove the excess acetone. The resulting
aqueous mixture was extracted with EtOAc twice. The combined
organic portion was dried over sodium sulfate and concentrated to
afford the title compound as white solid. (4.22 g) LC/MS, ESI-MS:
non-ionizable, RT: 1.05 (Condition E).
Step 5. Synthesis of methyl
2-(4-(4-bromophenyl)-4-(hydroxymethyl)cyclohexylidene)acetate
##STR00045##
[0207] To a round bottom flask containing methanol (60 ml) was
added sodium hydride (0.78 g, 19.37 mmol) portion-wise at room
temperature under stirring, Trimethyl phosphonoacetate (2.58 ml,
17.88 mmol) was added drop-wise and the resulting mixture was
stirred at room temperature for 30 minutes. To this mixture was
then added 4-(4-bromophenyl)-4-(hydroxymethyl)cyclohexanone (4.22
g, 14.90 mmol) in several portions and stirred at room temperature
for overnight. The reaction mixture was then quenched with a
saturated solution of ammonium chloride and subsequently
concentrated to remove excess methanol. The residue was taken in
saturated ammonium chloride solution and extracted twice with
EtOAc. The combined organic portion was dried over sodium sulfate
and concentrated to afford a crude product. Column purification
afforded the title compound as viscous oil (4.36 g, 86% yield).
LC/MS, ESI-MS: non-ionizable, RT: 1.32 (Condition E).
Step 6. Synthesis of methyl
2-(4-(4-bromophenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00046##
[0209] To a round bottom flask containing methyl
2-(4-(4-bromophenyl)-4-(hydroxymethyl)cyclohexylidene)acetate (4.36
g, 12.85 mmol) was added 1,4-dioxane (161 ml). The clear solution
was cooled in an ice-bath and sodium hydride (0.67 g, 16.75 mmol)
was then added portion-wise. After the addition, the mixture was
warmed to room temperature and stirred for 10 minutes. After 10
minutes of stirring, the content was heated to 100.degree. C. in an
oil bath (with a reflux condenser) for 30 minutes. The reaction
mixture was cooled to room temperature and quenched with a
saturated solution of ammonium chloride and subsequently
concentrated to remove excess dioxane. The residue was taken in
saturated ammonium chloride solution and extracted twice with
EtOAc. The combined organic portion was dried over sodium sulfate
and concentrated to afford a crude product. Column purification
afforded the title compound as white solid (3.4 g, 78% yield).
LC/MS, ESI-MS: 341.2 (M+H.sup.+), RT: 1.40 (Condition E).
Step 7. Synthesis of methyl
2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-1-yl)acetate
##STR00047##
[0211] To a round bottom flask containing methyl
2-(4-(4-bromophenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate (3.43 g,
10.11 mmol) was added bis(pinacolato)diboron (3.85 g, 15.17 mmol),
potassium acetate (2.98 g, 30.3 mmol) and anhydrous 1,4-dioxane (48
ml). The mixture was stirred and degassed with nitrogen at room
temperate for 15 minutes. PdCl.sub.2(dppf) (0.41 g, 0.51 mmol) was
then added and the mixture heated to 80.degree. C. for overnight.
The dark mixture was then cooled to room temperature and filtered
over a pad of celite. The resulting filtrate was concentrated and
column purified to afford the title compound as off-white solid
(3.87 g, 99% yield). LC/MS, ESI-MS(+): 387.4, RT: 1.48 (Condition
E).
Step 8. Synthesis of
methyl2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)-
acetate
##STR00048##
[0213] To a microwave vial containing methyl
2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-1-yl)acetate (100 mg, 0.26 mmol),
6-bromopyridin-3-amine (49 mg, 0.28 mmol) and
Pd.sub.2(PPh.sub.3).sub.4 (60 mg, 0.05 mmol) was added 1,4-dioxane
(1.3 ml) at room temperature. 2N sodium carbonate (0.39 ml, 0.78
mmol) was added and the mixture was subjected to microwave
radiation at 120.degree. C. for 50 minutes. The resulting mixture
was concentrated to dryness and taken up in EtOAc, the slurry was
dried over sodium sulfate and filtered through a pad of celite. The
filtrate was concentrated and purified by column chromatography to
afford the title compound (120 mg, 65% yield). LC/MS, ESI-MS(+):
353.4, RT: 1.12 (Condition E).
Step 9. Synthesis of methyl
2-(4-(4-(5-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)pyridin-2-yl)phenyl)-2-
-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00049##
[0215] To a reaction vial containing
methyl2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)-
acetate (120 mg, 0.23 mmol) was added dichloromethane (2 ml) at
room temperature. 1,1'-thiocarbonyldipyridin-2(1H)-one (59 mg, 0.26
mmol) was added and stirred at room temperature for 1 hour to
afford a bright orange mixture. Pivalohydrazine (40 mg, 0.35 mmol)
was then added in one portion and the mixture stirred at room
temperature for another hour. 1-ethyl-3-(3-dimethylaminopropyl)
carbodiimide (80 mg, 0.42 mmol) was added and the mixture was
stirred at room temperature for overnight. The content was then
concentrated to dryness and the resulting residue was taken up in
water to form a nice slurry. The slurry was filtered and washed
with water, dried to afford the title compound as off-white solid
(55 mg, 50% yield). LC/MS, ESI-MS(+): 477.4, RT: 1.31 (Condition
E).
Step 10. Synthesis of
2-(4-(4-(5-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)pyridin-2-yl)phenyl)-2-
-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00050##
[0217] To a reaction vial containing methyl
2-(4-(4-(5-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)pyridin-2-yl)phenyl)-2-
-oxabicyclo[2.2.2]octan-1-yl)acetate (55 mg, 0.12 mmol) was added
THF (1.5 ml) and a solution of 1N NaOH (0.35 ml, 0.35 mmol). The
mixture was stirred vigorously at room temperature for overnight.
The resulting mixture was concentrated to dryness and the residue
was taken up in water. The mixture was acidified with 1N HCl to a
pH of between 3 and 4 affording a thick residue. The residue was
concentrated to dryness and taken up in MeOH to form fine slurry.
The slurry was filtered and washed with small amount of MeOH and
water to afford the title compound as white solid after drying (36
mg, 67% yield). HR/MS (M+H).sup.+. found 463.2354, calc. 463.2345.
RT: 5.76 (Condition Z). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.09 (br. s, 1H) 10.78 (br. s, 1H) 8.76 (d, J=2.78 Hz, 1H)
8.04-8.13 (m, 1H) 7.90-8.01 (m, 3H) 7.41 (d, J=8.59 Hz, 2H) 3.90
(s, 2H) 2.29 (s, 2H) 1.82-2.08 (m, 8H) 1.36 (s, 9H)
Example 2
2-(4-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)--
2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
2-(4-(4-(5-(5-cyclobutyl-1,3,4-oxadiazol-2-ylamino)pyridin-2-yl)phenyl)-2-
-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00051##
[0219] The title compound was prepared analogous to Example1, Step
9 starting from
methyl2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)-
acetate (35 mg, 0.10 mmol) and cyclobutylcarbohydrazide (17 mg,
0.15 mmol). Column purification afforded the title compound as
white solid after drying (38 mg, 81% yield). LC/MS, ESI-MS(+):
475.25, RT: 1.14 (Condition E).
Step 2. Synthesis of
2-(4-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00052##
[0221] To a mixture of methyl
2-(4-(4-(5-(5-cyclobutyl-1,3,4-oxadiazol-2-ylamino)pyridin-2-yl)phenyl)-2-
-oxabicyclo[2.2.2]octan-1-yl)acetate (38 mg, 0.08 mmol) and THF (1
ml) was added NaOH (0.24 ml, 0.24 mmol). The mixture was stirred at
room temperature for 7 hour. The resulting content was concentrated
and purified on prep-HPLC to afford the title compound as white
solid after drying (15 mg, 41% yield). HR/MS (M+H).sup.+. found
461.2188, calc. 461.2184. RT: 5.63 (Condition Z). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 1.85-2.13 (m, 10H) 2.27-2.40 (m, 6H)
3.69 (quind, J=8.43, 8.43, 8.43, 8.43, 0.88 Hz, 1H) 3.91 (s, 2H)
7.41 (d, J=8.59 Hz, 2H) 7.90-8.01 (m, 3H) 8.10 (dd, J=8.59, 2.78
Hz, 1H) 8.75 (d, J=3.03 Hz, 1H)
Example 3
2-(4-(4-(5-((5-cyclobutyl-1,3,4-thiadiazol-2-yl)amino)pyridin-2-yl)phenyl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of ethyl/methyl
2-(4-(4-(5-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyridin-2-yl)phenyl)--
2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00053##
[0223] To a solution of
methyl2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)-
acetate (57 mg, 0.16 mmol) in DCM (0.5 ml) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (39 mg, 0.17 mmol) and the
mixture was stirred at room temperature. After 30 minutes of
stirring, cyclobutylcarbohydrazide (28 mg, 0.24 mmol) was added and
the reaction was stirred at room temperature for 2 hour. Afterward,
a solution of concentrated H.sub.2SO.sub.4 (0.05 ml, 0.94 mmol) was
added and the mixture stirred at room temperature for 3 hour. The
mixture was taken up in EtOAc and washed with NaHCO.sub.3, then
brine. The organic portion was dried over sodium MgSO.sub.4,
filtered and concentrated to afford a crude product. Column
purification afforded the title compound containing approximately
25% of ethyl ester (total yield 33 mg,). The mixture was used in
the next step without additional purification. LC/MS, ESI-MS(+):
Methyl ester, 491.7, RT: 1.20 (Condition E). Ethyl ester, 505.6,
RT: 1.26 (Condition E).
Step 2. Synthesis of
2-(4-(4-(5-((5-cyclobutyl-1,3,4-thiadiazol-2-yl)amino)pyridin-2-yl)phenyl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00054##
[0225] The title compound was prepared analogous to Example 2, Step
2 starting from ethyl/methyl
2-(4-(4-(5-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyridin-2-yl)phenyl)--
2-oxabicyclo[2.2.2]octan-1-yl)acetate (33 mg, 0.07 mmol) and 1N
NaOH (0.20 ml, 0.20 mmol) solution. Prep-HPLC purification afforded
the title compound as white solid after drying (18 mg, 56% yield).
HR/MS (M+H).sup.+. found 477.1961, calc. 477.1955. RT: 5.93
(Condition Z).). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
1.85-2.08 (m, 10H) 2.30 (s, 4H) 2.35-2.44 (m, 2H) 3.84 (quind,
J=8.49, 8.49, 8.49, 8.49, 0.88 Hz, 1H) 3.90 (s, 2H) 7.41 (d, J=8.59
Hz, 2H) 7.93 (d, J=8.59 Hz, 1H) 7.97 (d, J=8.59 Hz, 2H) 8.25 (dd,
J=8.72, 2.65 Hz, 1H) 8.77 (d, J=2.78 Hz, 1H)
Example 4
2-(4-(4-(2-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyrimidin-5-yl)phenyl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
2-(4-(4-(2-aminopyrimidin-5-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acet-
ate
##STR00055##
[0227] To a reaction vial containing methyl
2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-1-yl)acetate (200 mg, 0.52 mmol),
5-bromopyrimidin-2-amine (108 mg, 0.62 mmol) and K.sub.3PO.sub.4
(132 mg, 0.62 mmol) was added dimethoxyethane (4.8 ml). The mixture
was stirred at room temperature and degassed after the addition of
EtOH (1.6 ml) and water (0.64 ml). PdCl.sub.2(dppf) DCM adduct (21
mg, 0.03 mmol) was added and the mixture was heated to 80 C in an
oil bath for 3 hours. The reaction mixture was then concentrated to
dryness and taken up in DCM. The slurry was dried over sodium
sulfate and filtered through a pad of celite. The filtrate was
concentrated and purified by column chromatography to afford the
title compound (193 mg, 84% yield). LC/MS, ESI-MS(+): 354.3, RT:
1.06 (Condition E).
Step 2. Synthesis of methyl
2-(4-(4-(2-(2-cyclobutanecarbonyl)hydrazinecarbothioamido)pyrimidin-5-yl)-
phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00056##
[0229] To a reaction vial containing methyl
2-(4-(4-(2-aminopyrimidin-5-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acet-
ate (40 mg, 0.11 mmol) was added dichloromethane (1 ml) at room
temperature. 1,1'-thiocarbonyldipyridin-2(1H)-one (29 mg, 0.12
mmol) was added and the resulting mixture was heated to 50.degree.
C. for overnight. The mixture was cool down to room temperature and
cyclobutanecarbohydrazide (19 mg, 0.17 mmol) was then added and
stirred at room temperature for another hour. The content was then
concentrated to dryness and the resulting residue was taken up in
MeOH to form a nice slurry. The slurry was filtered and washed with
small amount of MeOH to afford the title compound as off-white
solid after drying (48 mg, 83% yield). LC/MS, ESI-MS(+): 510.4, RT:
1.26 (Condition E)
Step 3. Synthesis of ethyl/methyl
2-(4-(4-(2-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyrimidin-5-yl)phenyl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00057##
[0231] To a reaction vial containing methyl
2-(4-(4-(2-(2-cyclobutanecarbonyl)
hydrazinecarbothioamido)pyrimidin-5-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-
-yl)acetate (49 mg, 0.10 mmol) was added EtOH (0.6 ml) at room
temperature. A solution of concentrated H.sub.2SO.sub.4 (0.03 ml,
0.56 mmol) was added and the mixture heated to 80 C for 1 hour. The
resulting mixture was cooled to room temperature and quenched with
a solution of saturated NaHCO.sub.3 dropwise. The content was
concentrated to dryness and subsequently taken up in DCM and dried
over sodium sulfate. The mixture was filtered and concentrated to
afford a crude residue. Column purification afforded the title
compound containing approximately 18% of methyl ester (total yield
41 mg,). The mixture was used in the next step without additional
purification. LC/MS, ESI-MS(+): Methyl ester, 492.2, RT: 1.42
(Condition E). Ethyl ester, 506.2, RT: 1.50 (Condition E).
Step 4. Synthesis of
2-(4-(4-(2-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyrimidin-5-yl)phenyl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00058##
[0233] To a reaction vial containing a mixture of ethyl/methyl
2-(4-(4-(2-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyrimidin-5-yl)phenyl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetate (41 mg, 0.08 mmol) was
added THF (1 ml) and MeOH (1 ml) at room temperature. NaOH (0.24
ml, 0.24 mmol) was added and the mixture stirred at room
temperature for overnight. The reaction mixture was then
concentrated to dryness and taken up in water. The mixture was
acidified with 1N HCl to a pH of between 3 and 4 affording thick
slurry. The slurry is filtered and washed with small amount of
water to afford the title compound as white solid after drying (37
mg, 97% yield). HR/MS (M+H).sup.+. found 478.1901, calc. 478.1913.
RT: 2.68 (Condition L). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.16 (br. s, 2H) 8.95 (s, 2H) 7.69 (d, J=8.59 Hz, 2H) 7.44 (d,
J=8.59 Hz, 2H) 3.90 (s, 3H) 2.37-2.47 (m, 2H) 2.24-2.36 (m, 4H)
1.82-2.12 (m, 10H).
Example 5
2-(4-(4-(6-((5-cyclobutyl-1,3,4-thiadiazol-2-yl)amino)pyridin-3-yl)phenyl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
2-(4-(4-(6-aminopyridin-3-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e
##STR00059##
[0235] The title compound was prepared analogous to Example 4, Step
1 starting from methyl
2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-1-yl)acetate (200 mg, 0.52 mmol) and
5-iodopyridin-2-amine (137 mg, 0.62 mmol). The reaction was
performed at 80.degree. C. for overnight. After workup and column
purification afforded the title compound as off-white solid after
drying (133 mg, 58% yield). LC/MS, ESI-MS(+):353.3, RT: 1.12
(Condition E).
Step 2. Synthesis of ethyl/methyl
2-(4-(4-(6-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyridin-3-yl)phenyl)--
2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00060##
[0237] To a reaction vial containing methyl
2-(4-(4-(6-aminopyridin-3-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (40 mg, 0.11 mmol) and DCM (1 ml) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (29 mg, 0.12 mmol). The
mixture was stirred at room temperature for 1 hour.
Cyclobutanecarbohydrazine (19 mg, 0.17 mmol) was then added and the
reaction stirred at room temperature for another hour. The mixture
was then concentrated to dryness, EtOH (1 ml), followed by a
solution of concentrated H.sub.2SO.sub.4 (0.04 ml, 0.66 mmol) was
added. The mixture was heated to 80.degree. C. for 1 hour. The
resulting mixture was cooled to room temperature and quenched with
a solution of saturated NaHCO.sub.3 dropwise. The content was
concentrated to dryness and subsequently taken up in DCM and dried
over sodium sulfate. The mixture was filtered and concentrated to
afford a crude residue. Column purification afforded the title
compound containing approximately 47% of methyl ester (total yield
48 mg,). The mixture was used in the next step without additional
purification. LC/MS, ESI-MS(+): Methyl ester, 491.2, RT: 2.88
(Condition M). Ethyl ester, 505.2, RT: 3.06 (Condition M).
Step 3. Synthesis of
2-(4-(4-(6-((5-cyclobutyl-1,3,4-thiadiazol-2-yl)amino)pyridin-3-yl)phenyl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00061##
[0239] To a reaction vial containing a mixture of ethyl/methyl
2-(4-(4-(6-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)pyridin-3-yl)phenyl)--
2-oxabicyclo[2.2.2]octan-1-yl)acetate (48 mg, 0.10 mmol) was added
THF (0.5 ml) and MeOH (0.5 ml) at room temperature. 1N NaOH (0.29
ml, 0.29 mmol) was added and the mixture was stirred at RT for
overnight. The mixture was concentrated to dryness and taken in
water to form a slurry. HCl was added adjusting the pH to 3 and 4.
The resulting slurry was filtered, washed with water to afford the
title compound as white solid after drying (36.8 mg, 79% yield).
HR/MS (M+H).sup.+. found 477.1949, calc. 477.1960. RT: 2.83
(Condition L). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
10.95-12.82 (m, 2H) 8.58 (d, J=2.27 Hz, 1H) 7.91-8.14 (m, 1H) 7.62
(d, J=8.34 Hz, 2H) 7.41 (d, J=8.59 Hz, 2H) 7.14 (d, J=8.59 Hz, 1H)
3.79-3.95 (m, 3H) 2.24-2.47 (m, 6H) 1.82-2.13 (m, 10H).
Example 6
2-(4-(4'-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
2-(4-(4'-nitrobiphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00062##
[0241] To a round bottom flask containing methyl
2-(4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-1-yl)acetate (800 m g, 2.07 mmol),
1-iodo-4-nitrobenzene (619 mg, 2.49 mmol) and K.sub.3PO.sub.4 (528
mg, 2.49 mmol) was added dimethoxyethane (24 ml) at room
temperature. The mixture was stirred at room temperature and
degassed after the addition of EtOH (8 ml) and water (3.2 ml).
PdCl.sub.2(dppf)dichloromethane adduct (85 mg, 0.10 mmol) was added
and the mixture was heated to 80 C in an oil bath for overnight.
The reaction mixture was then concentrated to dryness and taken up
in DCM. The slurry was dried over sodium sulfate and filtered
through a pad of celite. The filtrate was concentrated and purified
by column chromatography affording the title compound as off-white
solid (624 mg, 79% yield). LC/MS, ESI-MS(+): 382.4, RT: 1.49
(Condition E).
Step 2. Synthesis of methyl
2-(4-(4'-aminobiphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)
acetate
##STR00063##
[0243] To a round bottom flask containing methyl
2-(4-(4'-nitrobiphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate
(716 mg, 1.88 mmol) was added Pd(OH).sub.2 on activated carbon (395
mg, 2.82 mmol). EtOAc (15 ml) and MeOH (3 ml) were added and the
mixture was stirred under 1 atm. of hydrogen at room temperature
for 2 hours. The resulting mixture was filtered through a pad of
celite and concentrated to afford a crude residue. Column
purification affords the title compound as off-white solid after
drying (595 mg, 90% yield). LC/MS, ESI-MS(+): 352.3, RT: 1.29
(Condition E).
Step 3. Synthesis of methyl 2-(4-(4'-(2-(cyclobutanecarbonyl)
hydrazinecarbothioamido)
biphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl) acetate
##STR00064##
[0245] The title compound was prepared analogous to Example 4, Step
2, starting from methyl
2-(4-(4'-aminobiphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl) acetate
(500 mg, 1.48 mmol) with 1,1'-thiocarbonyldipyridin-2(1H)-one (379
mg, 1.63 mmol) and cyclobutanecarbohydrazide (254 mg, 2.22 mmol).
Water titration of the crude residue affords the title compound as
off-white solid after drying (696 mg, 93% yield). LC/MS, ESI-MS(+):
508.4, RT: 1.29 (Condition E).
Step 4. Synthesis of ethyl/methyl
2-(4-(4'-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)biphenyl-4-yl)-2-oxabic-
yclo[2.2.2]octan-1-yl)acetate
##STR00065##
[0247] The title compound was prepared analogous to Example 4, Step
3 starting from methyl 2-(4-(4'-(2-(cyclobutanecarbonyl)
hydrazinecarbothioamido)
biphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl) acetate (300 mg, 0.59
mmol) with concentrated H.sub.2SO.sub.4 (0.19 ml, 3.43 mmol) in
EtOH. Column purification affords the title compound containing
approximately 24% of methyl ester (total yield 235 mg,). The
mixture was used in the next step without additional purification.
LC/MS, ESI-MS(+): Methyl ester, 490.4, RT: 1.47 (Condition E).
Ethyl ester, 504.4, RT: 1.52 (Condition E).
Step 5. Synthesis of
2-(4-(4'-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)biphenyl-4-yl)-2-oxabic-
yclo[2.2.2]octan-1-yl)acetic acid
##STR00066##
[0249] To a reaction vial containing a mixture of ethyl/methyl
2-(4-(4'-(5-cyclobutyl-1,3,4-thiadiazol-2-ylamino)biphenyl-4-yl)-2-oxabic-
yclo[2.2.2]octan-1-yl)acetate (235 mg, 0.47 mmol) was added THF (2
ml) and MeOH (2 ml) at room temperature. 1N NaOH (1.4 ml, 1.4 mmol)
was added and the mixture was stirred at RT for overnight. The
mixture was concentrated to dryness and taken in water to form a
nice slurry. The slurry was then filtered and washed with water to
afford the title compound as sodium salt. To a round bottom flask
containing the sodium salt was added a mixture of 1:1 acetonitrile
and water (total volume 10 ml) at room temperature. The mixture was
acidified with 1N HCl to a pH of between 3 and 4 affording a thick
slurry. The mixture was then concentrated to remove excess
acetonitrile and filtered. The filter cake was washed with water to
afford the title compound as white solid after drying (178 mg, 80%
yield). HR/MS (M+H).sup.+. found 476.2014, calc. 476.2008. RT: 2.89
(Condition L). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
10.48 (br. s, 1H) 7.68 (d, J=8.84 Hz, 2H) 7.62 (d, J=8.84 Hz, 2H)
7.57 (d, J=8.59 Hz, 2H) 7.37 (d, J=8.34 Hz, 2H) 3.87 (s, 2H)
3.79-3.85 (m, 1H) 2.20-2.45 (m, 4H) 1.76-2.19 (m, 12H)
Example 7
2-(4-(4'-(5-cyclobutyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicyc-
lo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
2-(4-(4'-(5-cyclobutyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetate
##STR00067##
[0251] To a reaction vial containing methyl
2-(4-(4'-(2-(cyclobutanecarbonyl) hydrazinecarbothioamido)
biphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl) acetate (300 mg, 0.59
mmol) was added DCM (6 ml) at room temperature.
1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (204 mg, 1.06 mmol)
was added and the mixture was stirred at room temperature for
overnight. The reaction mixture was purified directly via column
chromatography affording the title compound as off-white solid
after drying (252 mg, 90% yield). LC/MS, ESI-MS(+): 474.4, RT: 1.40
(Condition E).
Step 2. Synthesis of
2-(4-(4'-(5-cyclobutyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetic acid
##STR00068##
[0253] The title compound was prepared analogous to Example 4, Step
4, starting from methyl
2-(4-(4'-(5-cyclobutyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetate (252 mg, 0.53 mmol) and NaOH (1.60 ml,
1.60 mmol). After acidification with 1N HCl, the slurry was
filtered and washed with water to afford the title compound as
white solid after drying (226 mg, 92% yield). HR/MS (M+H).sup.+.
found 460.2221, calc. 476.2236. RT: 2.68 (Condition L). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 10.53 (s, 1H) 7.62 (s, 4H) 7.57
(d, J=8.59 Hz, 2H) 7.38 (d, J=8.59 Hz, 2H) 3.89 (s, 2H) 3.62-3.73
(m, 1H) 2.22-2.41 (m, 6H) 1.82-2.11 (m, 10H)
Example 8
2-(4-(4'-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicyc-
lo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
2-(4-(4'-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetate
##STR00069##
[0255] The title compound was prepared analogous to Example 1, Step
9 starting from methyl
2-(4-(4'-aminobiphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl) acetate
(95 mg, 0.28 mmol). The crude residue was triturated in MeOH and
filtered to afford the title compound as off-white solid after
drying (117 mg, 87% yield). LC/MS, ESI-MS(+): 476.4, RT: 1.42
(Condition E).
Step 2. Synthesis of
2-(4-(4'-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetic acid
##STR00070##
[0257] The title compound was prepared analogous to Example 4, Step
4 starting from methyl
2-(4-(4'-(5-tert-butyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetate (82 mg, 0.17 mmol) and NaOH (0.52 ml,
0.52 mmol). After acidification with 1N HCl, the slurry was
filtered and washed with water to afford the title compound as
white solid after drying (75 mg, 95% yield). HR/MS (M+H).sup.+.
found 462.2396, calc. 462.2392. RT: 2.75 (Condition L). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 12.07 (s, 1H) 10.51 (s, 1H)
7.62 (s, 4H) 7.58 (d, J=8.34 Hz, 2H) 7.38 (d, J=8.59 Hz, 2H) 3.90
(s, 2H) 2.33 (s, 2H) 1.82-2.07 (m, 8H) 1.36 (s, 9H)
Example 9
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)--
2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of methyl
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00071##
[0259] The title compound was prepared analogous to Example 1, Step
9 starting from methyl
2-(4-(4'-aminobiphenyl-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl) acetate
(335 mg, 0.95 mmol). The crude residue was purified by column
chromatography to afford the title compound as off-white solid
after drying (117 mg, 87% yield). LC/MS, ESI-MS(+): 460.2, RT: 1.34
(Condition W).
Step 2. Synthesis of
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid sodium salt
##STR00072##
[0261] The title compound was prepared analogous to Example 4, Step
5 starting from methyl
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetate (413 mg, 0.90 mmol) and
NaOH (2.70 ml, 2.70 mmol). After completion of the reaction, the
mixture was concentrated to dryness and taken up in water. The
resulting slurry was filtered and washed with water to afford the
title compound as white solid after drying (392 mg, 93% yield).
HR/MS (M+H).sup.+. found 446.2077, calc. 446.2080. RT: 1.54
(Condition L). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
10.54 (br. s., 1H) 7.58-7.64 (m, 4H) 7.55 (d, J=8.34 Hz, 2H) 7.36
(d, J=8.34 Hz, 2H) 3.84 (s, 2H) 2.16-2.27 (m, 2H) 2.13 (tt, J=8.37,
5.02 Hz, 1H) 2.00 (s, 2H) 1.88-1.99 (m, 2H) 1.69-1.88 (m, 4H)
1.03-1.10 (m, 2H) 0.91-0.98 (m, 2H)
Step 3. Synthesis of
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00073##
[0263] To a reaction vial contaminions
2-(4-(4'-((5-cyclopropyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid sodium salt (195 mg,
0.42 mmol) was added water (5 ml). The mixture was stirred and
hydrochloric acid was added dropwise until the entire mixture
achieved a pH between 3 and 4. The resulting slurry was filtered,
washed with water to afford the title compound as white solid after
drying (183 mg, 99% yield). HR/MS (M+H).sup.+. found 446.2070,
calc. 446.2080. RT: 1.66 (Condition L). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.01 (br. s, 1H) 10.43 (s, 1H) 7.52-7.67
(m, 6H) 7.38 (d, J=8.59 Hz, 2H) 3.89 (s, 2H) 2.32 (s, 2H) 2.08-2.19
(m, 1H) 1.79-2.07 (m, 8H) 1.02-1.12 (m, 2H) 0.91-0.99 (m, 2H).
Example 10
2-(4-(4'-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)-[1,1-biphenyl-
]-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of
2-methyl-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-5-(tri-
fluoromethyl)oxazole-4-carboxamide
##STR00074##
[0265] To a stirred solution of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) aniline (420 mg,
1.917 mmol) in anhydrous DMF (9.6 mL) was added
2-methyl-5-(trifluoromethyl)oxazole-4-carboxylic acid (449 mg,
2.300 mmol) followed by DIPEA (1.0 mL, 5.75 mmol) and the mixture
was stirred at room temperature for 10 minutes. HATU (875 mg, 2.300
mmol) was added and the mixture stirred at room temperature for 16
hours. The mixture was taken up in 1:1 mixture of brine and water
and extracted with EtOAc. The organic portion was dried over sodium
sulfate, filtered and concentrated to afford a brown oil, which was
purified by flash column chromatography to afford the title
compound as pale yellow solid after drying (360 mg, 47% yield).
LC/MS, ESI-MS (+): 397.1, RT: 1.49 (Condition E).
Step 2. Synthesis of methyl
2-(4-(4'-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)-[1,1'-biphen-
yl]-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00075##
[0267] To a mixture of
2-methyl-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-5-(tri-
fluoromethyl)oxazole-4-carboxamide (300 mg, 0.757 mmol) and methyl
2-(4-(4-bromophenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate (257 mg,
0.757 mmol) in dioxane (3 ml) was added H.sub.2O (0.33 ml) at room
temperature. CsF (345 mg, 2.272 mmol) and Pd(amphos)Cl.sub.2 (53.6
mg, 0.076 mmol) were added and the mixture was degassed several
times before being heated to 90.degree. C. for overnight. The
reaction mixture was cooled to room temperature and directly
purified by column chromatography to afford the title compound as
white solid after drying (200 mg, 50% yield). LC/MS, ESI-MS (+):
529.0, RT: 1.52 (Condition E).
Step 3. Synthesis of
2-(4-(4-(5-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)pyridin-2-y-
l)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00076##
[0269] To a solution of methyl
2-(4-(4'-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)biphenyl-4-yl-
)-2-oxabicyclo[2.2.2]octan-1-yl)acetate (200 mg, 0.378 mmol) in THF
(4 ml) at room temperature was added 1N LiOH (0.454 ml, 0.454
mmol). The mixture was stirred at room temperature for overnight.
Water (4 ml) was added and the mixture was quenched with 1N HCl
(0.454 ml, 0.454 mmol). The resulting slurry was stirred for 15
minutes and filtered. The solid was triturated with hot EtOAc and
filtered to afford the title compound as white solid after drying
(116 mg, 60% yield). HR/MS (M+H).sup.+. found 515.1788, calc.
515.1782. RT: 2.96 (Condition L). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.08 (s, 1H), 10.66 (s, 1H), 7.88 (d,
J=8.0 Hz, 2H), 7.62 (m, 4H), 7.39 (d, J=8.00 Hz, 2H), 3.95 (s, 2H),
2.65 (s, 3H), 2.33 (s, 3H), 2.01-1.81 (m, 8H).
Example 11
2-(4-(4'-(2-Ethyl-4-methyloxazole-5-carboxamido)biphenyl-4-yl)-2-oxabicycl-
o[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of
2-ethyl-4-methyl-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl-
)oxazole-5-carboxamide
##STR00077##
[0271] To a stirred solution of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) aniline (360 mg,
1.643 mmol) in anhydrous DMF (8.2 mL) was added
2-ethyl-4-methyloxazole-5-carboxylic acid (306 mg, 1.972 mmol),
followed by DIPEA (0.86 mL, 4.93 mmol) and the mixture was stirred
at room temperature for 10 minutes. HATU (750 mg, 1.972 mmol) was
added and the mixture was stirred at room temperature for 16 hour.
The mixture was taken up in 1:1 mixture of brine and water (40 mL)
and extracted with EtOAc (60 mL). The organic portion was dried
over sodium sulfate, filtered and concentrated to afford a brown
oil, which was purified by flash column chromatography to afford
the title compound (305 mg, 52% yield) as a white solid. LC/MS,
ESI-MS (+): 357.1, RT: 1.35 (Condition E).
Step 2. Synthesis of methyl
2-(4-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1'-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetate
##STR00078##
[0273] To a solution of
2-ethyl-4-methyl-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl-
)oxazole-5-carboxamide (200 mg, 0.561 mmol) and methyl
2-(4-(4-bromophenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetate (190 mg,
0.561 mmol) in dioxane (3 ml) was added H.sub.2O (0.33 ml). CsF
(256 mg, 1.684 mmol) and Pd(amphos)Cl.sub.2 (39.8 mg, 0.056 mmol)
were then added and the mixture was degassed before being heated at
90.degree. C. for overnight. The reaction mixture was then cooled
to room temperature and directly purified by column chromatography
to afford the title compound as white solid after drying (159 mg,
58% yield). LC/MS, ESI-MS (+): 489.0, RT: 1.40 (Condition E).
Step 3. Synthesis of
2-(4-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1-biphenyl]-4-yl)-2-o-
xabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00079##
[0275] To a solution of
2-(4-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1'-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetate (159 mg, 0.325 mmol) in THF (3
ml) at room temperature was added 1N LiOH (0.391 ml, 0.391 mmol).
The mixture was stirred at room temperature overnight. The mixture
was quenched with 1N HCl (0.391 ml, 0.391 mmol) and diluted with
H.sub.2O. The THF was removed in vacuo to afford off-white slurry.
The slurry was stirred for 15 min then filtered. The filter cake
was washed with H.sub.2O, heptane, and dried to afford the title
compound as off-white sold (135 mg, 87% yield).
[0276] HR/MS (M+H).sup.+. found 475.2222, calc. 475.2233. RT: 2.63
(Condition L). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
12.08 (s, 1H), 10.18 (s, 1H), 7.82 (d, J=8.0 Hz, 2H), 7.62 (m, 4H),
7.39 (d, J=8.0 Hz, 2H), 3.90 (s, 2H), 2.85 (q, J=8.0 Hz, 2H), 2.49
(s, 3H), 2.33 (s, 2H), 2.06-1.85 (m, 8H), 1.33 (t, J=8.0 Hz,
3H).
Example 12
2-(4-(4-(5-(2-methyl-5-(trifluoromethyl)oxazole-4-carboxamido)pyridin-2-yl-
)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00080##
[0278] To a solution of methyl
2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (212 mg, 0.602 mmol) in DMF (3 ml) at rt under N.sub.2 was added
2-methyl-5-(trifluoromethyl)oxazole-4-carboxylic acid (141 mg,
0.722 mmol) and Et.sub.3N (0.167 ml, 1.203 mmol. The mixture was
stirred for 5 minutes before the addition of HATU (274 mg, 0.722
mmol) at room temperature. Upon completion of reaction, the mixture
was diluted with H.sub.2O (25 ml) and extracted with EtOAc (50 ml).
The organic portion was washed with brine, dried over sodium
sulfate, filtered and concentrated to afford the crude methyl
ester. LC/MS, ESI-MS (+): 530.0, RT: 1.37 (Condition E). The crude
methylester was suspended in THF (4 ml) and treated with 1N LiOH
(0.722 ml, 0.722 mmol). Upon completion of reaction, the mixture
was diluted with H.sub.2O and brought to pH 5 with acetic acid. The
resulting slurry was filtered, washed with H.sub.2O and dried at
80.degree. C. for several hours. The crude solid was triturated
with hot EtOH, AcN, and Et.sub.2O to give the title compound as
off-white solid after drying (65 mg, 19% yield). HR/MS (M+H).sup.+.
found 516.1749, calc. 516.1735, RT: 2.76 (Condition L). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 12.08 1 (s, 1H), 10.92 (s, 1H),
9.03 (s, 1H), 8.30 (d, J=8.0 Hz, 1H), 7.99 (d, J=16.0 Hz, 2H), 7.94
(d, J=16.0 Hz, 1H), 7.44 (d, J=8.00 Hz, 2H), 3.91 (s, 2H), 2.63 (s,
3H), 2.33 (s, 2H), 2.08-1.85 (m, 8H).
Example 13
2-(4-(4-(5-(2-ethyl-4-methyloxazole-5-carboxamido)pyridin-2-yl)phenyl)-2-o-
xabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00081##
[0280] The title compound was prepared analogous to Example 12,
starting from methyl
2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (114 mg, 0.32 mmol) and 2-ethyl-4-methyloxazole-5-carboxylic acid
(60.2 mg, 0.39 mmol). The crude product was purified on prep-HPLC
to afford the title compound as off-white solid after drying (28
mg, 17% yield). HR/MS (M+H).sup.+. found 476.2166, calc. 476.2185,
RT: 2.76 (Condition L). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 10.55 (s, 1H), 8.98 (s, 1H), 8.24 (d, J=8.0 Hz, 1H), 8.01 (d,
J=12.0 Hz, 2H), 7.95 (d, J=12.0 Hz, 1H), 7.43 (d, J=8.0 Hz, 2H),
3.91 (s, 2H), 2.86 (q, J=8.0 Hz, 2H), 2.36 (s, 2H), 2.25 (s, 2H),
2.09-1.90 (m, 8H), 1.36 (t, J=8.0 Hz, 3H).
Example 14
2-(4-(4-(5-((5-(tert-butyl)oxazol-2-yl)amino)pyridin-2-yl)phenyl)-2-oxabic-
yclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of Benzyl
2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e
##STR00082##
[0282] To a mixture of methyl
2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (286 mg, 0.812 mmol) in neat benzyl alcohol (6 mL, 57.9 mmol) was
added titanium(IV) isopropoxide (1.5 mL, 5.12 mmol) and 4 A
molecular sieves (5 g, 0.812 mmol). The reaction mixture was heated
at 130.degree. C. overnight. The reaction was quenched by saturated
NaHCO3 and was extracted with EtOAc. The combined organic layer was
filtered and washed with brine and dried over anhydrous sodium
sulfate, filtered and concentrated. The residue was purified by
prep-HPLC (15 to 90% ACN-water (0.1% NH4OH) with X-bridge C18
column) to give the title compound after drying (150 mg, 43%
yield). LC/MS, ESI-MS (+): 429.1, RT: 1.41 (Condition E).
Step 2. Synthesis of benyl
2-(4-(4-(5-((5-(tert-butyl)oxazol-2-yl)amino)pyridin-2-yl)phenyl)-2-oxabi-
cyclo[2.2.2]octan-1-yl)acetate
##STR00083##
[0284] To a solution of benzyl
2-(4-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (150 mg, 0.350 mmol) in DCM (6 ml) at room temperature was added
1,1'-thiocarbonyldi-2(1H)-pyridone (85 mg, 0.368 mmol) and the
reaction was stirred at room temperature for 0.5 hr. To the mixture
was added 1-amino-3,3-dimethylbutan-2-one (80 mg, 0.525 mmol) and
DIPEA (0.110 ml, 0.630 mmol) and the mixture was stirred at
40.degree. C. or until the reaction was completed. To the mixture
was added EDC.HCl (268 mg, 1.400 mmol) and the reaction was stirred
at 40.degree. C. overnight. The reaction mixture was concentrated
and purified by flash chromatography (10 to 50% EtOAc/Heptane) to
give the title compound (100 mg, 51.8% yield). LC/MS, ESI-MS(+) m/z
552.1, RT: 1.73 (Condition R).
Step 3. Synthesis of
2-(4-(4-(5-((5-(tert-butyl)oxazol-2-yl)amino)pyridin-2-yl)phenyl)-2-oxabi-
cyclo[2.2.2]octan-1-yl)acetic acid
##STR00084##
[0286] Benyl
2-(4-(4-(5-((5-(tert-butyl)oxazol-2-yl)amino)pyridin-2-yl)phenyl)-2-oxabi-
cyclo[2.2.2]octan-1-yl)acetate (100 mg, 0.181 mmol) was dissolved
in EtOAc/THF and hydrogenated with 10% Pd(OH).sub.2/C under H.sub.2
balloon for 3 hr. The reaction mixture was filtered and the
filtrate was concentrated. The residue was purified by prep-HPLC to
give the title compound (28 mg, 31.5% yield). HR/MS [M+H].sup.+.
found 462.2390, calc. 462.2393. RT: 2.83 (Condition L). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 1.26 (s, 9H), 1.86-2.03 (m,
8H), 2.32 (s, 2H), 3.90 (s, 2H), 6.60 (s, 1H), 7.39 (d, J=8.59 Hz,
2H), 7.87 (d, J=8.59 Hz, 1H), 7.95 (d, J=8.59 Hz, 2H), 8.15 (dd,
J=8.84, 2.78 Hz, 1H), 8.77 (d, J=2.78 Hz, 1H), 10.36 (br. s.,
1H).
Example 15
2-(4-(4'-((5-(tert-butyl)oxazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-2-oxabicy-
clo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of benzyl
2-(4-(4'-amino-[1,1'-biphenyl]-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e
##STR00085##
[0288] To a mixture of methyl
2-(4-(4'-amino-[1,1'-biphenyl]-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (0.323 g, 0.918 mmol) in toluene (35 ml) was added benzyl alcohol
(2.377 ml, 22.95 mmol), titanium(IV) isopropoxide (0.807 ml, 2.75
mmol) and 4 A molecular sieves (5 g, 0.918 mmol). The reaction
mixture was heated at 120.degree. C. for 48 hours. The resulting
mixture was taken up in EtOAc and filtered. The filtrate was
subsequently washed with water, then brine. The organic portion
dried over anhydrous sodium sulfate, filtered and concentrated. The
residue was purified by flash chromatography (10 to 50%
EtOAc/Heptane) to give the title compound after drying (270 mg,
yield 68.8% yield). LC/MS, ESI-MS(+) m/z 428.1, RT 1.59 (Condition
R).
Step 2. Synthesis of benzyl
2-(4-(4'-((5-(tert-butyl)oxazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-2-oxabic-
yclo[2.2.2]octan-1-yl)acetate
##STR00086##
[0290] The title compound was prepared analogous to Example 14,
step 2 starting from benzyl
2-(4-(4'-amino-[1,1'-biphenyl]-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (240 mg, 0.32 mmol). After workup the crude product was purified
by column chromatography to give the title compound (200 mg, 89%
yield). LC/MS, ESI-MS(+) m/z 551.1, RT 1.82 (Condition R).
Step 3.
2-(4-(4'-((5-(tert-butyl)oxazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)--
2-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00087##
[0292] The title compound was prepared analogous to Example 14,
step 3 starting from benzyl
2-(4-(4'-((5-(tert-butyl)oxazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)-2-oxabi-
cyclo[2.2.2]octan-1-yl)acetate (200 mg, 0.36 mmol). After workup
the crude product was purified by prep-HPLC to give the title
compound (37 mg, 22% yield). HR/MS [M+H].sup.+. found 461.2427,
calc. 461.2440. RT: 2.85 (Condition L). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 1.26 (s, 9H), 1.82-2.07 (m, 8H), 2.33 (s,
2H), 3.89 (s, 2H), 6.58 (s, 1H), 7.37 (d, J=8.59 Hz, 2H), 7.51-7.61
(m, 4H), 7.61-7.70 (m, 2H), 10.16 (s, 1H).
Example 16
2-(4-(4'-((5-isobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-2-o-
xabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of
5-isobutyl-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,-
4-oxadiazol-2-amine
##STR00088##
[0294] To a stirred solution of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) aniline (200 mg,
0.91 mmol) in methylene chloride (4.5 ml) was added
1,1'-thiocarbonyldipyridin-2(1H)-one (212 mg, 0.91 mmol) and
stirred at room temperature for 1 hour. Upon complete formation of
the isothiocyanate, 3-methylbutanehydrazide (159 mg, 1.37 mmol) was
added to the mixture and stirred at room temperature for overnight.
EDC (315 mg, 1.64 mmol) was then added and the reaction stirred at
room temperature for 14 hours. The reaction was concentrated to
dryness and taken up in water to form a slurry. The slurry was
filtered and washed with water to afford the crude product, which
was purified by column chromatography to afford the title compound
as off-white solid after drying (217 mg, 69% yield). LC/MS,
ESI-MS(+) m/z 344.1, RT 1.45 (Condition R).
Step 2. Synthesis of methyl
2-(4-(4'-(5-isobutyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicycl-
o[2.2.2]octan-1-yl)acetate
##STR00089##
[0296] The title compound was prepared analogous to Example 10,
step 2 starting from
5-isobutyl-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,-
4-oxadiazol-2-amine (61 mg, 0.18 mmol). After workup the crude
product was purified by column chromatography to afford the title
compound after drying (56 mg, 79% yield). LC/MS, ESI-MS(+) m/z
476.1, RT 1.50 (Condition R).
Step 3. Synthesis of
2-(4-(4'-((5-isobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00090##
[0298] The title compound was prepared analogous to Example 4, Step
4 starting from methyl
2-(4-(4'-(5-isobutyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicycl-
o[2.2.2]octan-1-yl)acetate (56 mg, 0.12 mmol) and NaOH (0.35 ml,
0.35 mmol). After acidification with 1N HCl, the slurry was
filtered and washed with water to afford the title compound as
white solid after drying (52 mg, 97% yield). HR/MS (M+H).sup.+.
found 462.2399, calc. 462.2406. RT: 2.40 (Condition L). .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 10.02-10.86 (m, 1H) 7.62 (s,
5H) 7.53-7.59 (m, 2H) 7.37 (d, J=8.59 Hz, 2H) 3.85 (s, 2H) 2.65 (d,
J=7.07 Hz, 2H) 2.11-2.25 (m, 2H) 2.00-2.09 (m, 3H) 1.90-2.00 (m,
2H) 1.81 (d, J=13.39 Hz, 4H) 0.97 (d, J=6.82 Hz, 6H)
Example 17
2-(4-(4'-((5-neopentyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-1-yl)acetic acid
Step 1. Synthesis of 3,3-dimethylbutanehydrazide
##STR00091##
[0300] To a mixture of methyl 3,3-dimethylbutanoate (500 mg, 3.84
mmol) in EtOH (12 ml) was added hydrazine monohydrate (1.87 ml,
38.4 mmol). The mixture was heated to 80.degree. C. under reflux
for overnight. The reaction was cooled back down to room
temperature and concentrated to dryness affording a crude residue.
The residue was dried under high vacuum to afford the title
compound as crystalline solid (90 mg, 18% yield). .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 8.69-9.12 (s, 1H) 4.13 (br. s, 2H)
1.89 (s, 2H) 0.94 (s, 9H).
Step 2. Synthesis of benzyl
2-(4-(4'-(5-neopentyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicyc-
lo[2.2.2]octan-1-yl)acetate
##STR00092##
[0302] The title compound was prepared analogous to Example 1, Step
9 starting from benzyl
2-(4-(4'-amino-[1,1'-biphenyl]-4-yl)-2-oxabicyclo[2.2.2]octan-1-yl)acetat-
e (30 mg, 0.07 mmol) and 3,3-dimethylbutanehydrazide (14 mg, 0.11
mmol). The crude residue was purified by column chromatography to
afford the title compound as off-white solid after drying (34 mg,
86% yield). LC/MS, ESI-MS(+): 566.1, RT: 1.73 (Condition R).
Step 3. Synthesis of
2-(4-(4'-((5-neopentyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-2-
-oxabicyclo[2.2.2]octan-1-yl)acetic acid
##STR00093##
[0304] The title compound was prepared analogous to Example 14,
Step 3 starting from benzyl
2-(4-(4'-(5-neopentyl-1,3,4-oxadiazol-2-ylamino)biphenyl-4-yl)-2-oxabicyc-
lo[2.2.2]octan-1-yl)acetate (34 mg, 0.06 mmol) I). After
hydrogenation, the mixture was filtered and the filtercake was
washed with hot MeOH. The resulting filtrate was concentrated to
afford a crude solid. The crude material was triturated with DCM,
filtered and washed with a small amount of DCM to afford the title
compound as white solid after drying (10 mg, 34% yield). HR/MS
(M+H).sup.+. found 476.2535, calc. 462.2549. RT: 2.54 (Condition
L). .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.44-10.60 (m,
1H) 7.63 (s, 4H) 7.57 (d, J=8.59 Hz, 2H) 7.37 (d, J=8.59 Hz, 2H)
3.87 (s, 2H) 2.66 (s, 2H) 2.18 (s, 2H) 1.80-2.12 (m, 8H) 1.01 (s,
9H).
Example 18
{1-[4'-(5-tert-butyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bicy-
clo[2.2.2]oct-4-yl}-acetic acid
Step 1. Synthesis of
4-trifluoromethanesulfonyloxy-cyclohex-3-ene-1,1-dicarboxylic acid
diethyl ester
##STR00094##
[0306] An oven-dried flask charged with diisopropylamine (4.90 ml,
34.7 mmol) and tetrahydrofuran (30 ml), was cooled to 0.degree. C.
under nitrogen atmosphere. To the reaction mixture was added
n-butyl lithium (1.6M in hexanes, 21.67 ml, 34.7 mmol) dropwise and
then stirred at 0.degree. C. for 15 minutes. The reaction mixture
was cooled to -78.degree. C. and added the solution of
4-oxo-cyclohexane-1,1-dicarboxylic acid diethyl ester (7.0 g, 28.9
mmol) in tetrahydrofuran (15 ml) over 2 minutes. The reaction
mixture was stirred at -30.degree. C. for 30 minutes, and then
cooled to -78.degree. C., and followed by adding the solution of
2-[N,N-bis(trifluoromethylsulfonyl)amino]-5-chloropyridine (12.48
g, 31.8 mmol) in tetrahydrofuran (15 ml). The reaction mixture was
allowed to warm to room temperature and stirred for 2 hours. The
reaction mixture was concentrated and the resulting oil was
partitioned between ethyl acetate/heptane (3/1; 200 ml) and water
(100 ml). The layers were separated and the organic phase was
washed with water (5.times.50 ml), dried over Na.sub.2SO.sub.4, and
concentrated to afford the crude
4-trifluoromethanesulfonyloxy-cyclohex-3-ene-1,1-dicarboxylic acid
diethyl ester (12.8 g, 80% purity, 94% yield) as brown oil. .sup.1H
NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.27 (t, J=7.20 Hz, 6H)
2.25-2.35 (m, 2H) 2.43 (dd, J=6.19, 1.89 Hz, 2H) 2.72-2.80 (m, 2H)
4.21 (q, J=7.20 Hz, 4H) 5.69-5.82 (m, 1H).
Step 2. Synthesis of
4-(4-chloro-phenyl)-cyclohex-3-ene-1,1-dicarboxylic acid diethyl
ester
##STR00095##
[0308] A 100 ml heavy wall round bottom flask was charged with the
crude intermediate
4-trifluoromethanesulfonyloxy-cyclohex-3-ene-1,1-dicarboxylic acid
diethyl ester (4.0 g, 80% purity, 8.55 mmol, 1.0 equiv) from last
step, PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2 adduct (0.628 g, 0.769
mmol, 0.09 equiv), 4-chloro-phenylboric acid (2.67 g, 17.1 mmol,
2.0 equiv), tripotassium phosphate (4.54 g, 21.37 mmol, 2.5 equiv),
ethanol (8 ml), dimethoxyethane (32.0 ml) and t-butyl acetylene
(7.9 g, 97 mmol, 2.0 equiv). The flask was sealed, flushed with
nitrogen for three times and stirred at 80.degree. C. for 16 hours.
After cooled at room temperature, the reaction mixture was filtered
and washed with ethyl acetate (20 ml). The filtrate was
concentrated under vacuum and the residue was purified by
chromatography (heptanes/ethyl acetate 2/1) to give
4-(4-chloro-phenyl)-cyclohex-3-ene-1,1-dicarboxylic acid diethyl
ester (2.2 g, 76%) as colorless oil. .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 1.27 (t, J=8.00 Hz, 6H) 2.29 (t, J=6.38
Hz, 2H) 2.46 (td, J=6.25, 2.02 Hz, 2H) 2.65-2.85 (m, 2H) 4.21 (q,
J=8.00 Hz, 4H) 6.07 (m, 1H) 7.26 (s, 4H).
Step 3. Synthesis of
[4-(4-chloro-phenyl)-1-hydroxymethyl-cyclohex-3-enyl]-methanol
##STR00096##
[0310] An oven-dried flask under nitrogen was charged with
4-(4-chloro-phenyl)-cyclohex-3-ene-1,1-dicarboxylic acid diethyl
ester (6.5 g, 19.30 mmol) in tetrahydrofuran (80 ml), and at
-78.degree. C. was added lithium alumina hydride (30.9 ml, 30.9
mmol) in one portion. The mixture was allowed to warm to room
temperature and stirred for 2 hours. The reaction was quenched with
saturated aqueous sodium chloride solution (5 ml) at 0.degree. C.
The mixture was filtered and the filtrate was extracted with ethyl
acetate (3.times.100 ml), and the residue solid was washed with
acetone (100 ml). The organic solvents were combined, dried over
Na.sub.2SO.sub.4, and concentrated. The residue was purified by
chromatography (heptanes/acetone 1/2) to afford
[4-(4-chloro-phenyl)-1-hydroxymethyl-cyclohex-3-enyl]-methanol (4.7
g, 96%) as white solid. .sup.1H NMR (400 MHz, METHANOL-d.sub.4)
.delta. ppm 1.64-1.74 (m, 2H) 2.04 (d, J=3.66 Hz, 2H) 2.37-2.46 (m,
2H) 3.50 (td, J=10.80, 8.80 Hz, 4H) 6.03-6.11 (m, 1H) 7.27 (d,
J=8.40 Hz, 2H) 7.36 (d, J=8.40 Hz, 2H).
Step 4. Synthesis of
[6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-methanol
##STR00097##
[0312] To a stirred solution of
[4-(4-chloro-phenyl)-1-hydroxymethyl-cyclohex-3-enyl]-methanol (4.7
g, 18.60 mmol) in tetrahydrofuran (280 ml) at 0.degree. C., was
added the solution of N-bromosuccinimde (3.48 g, 19.53 mmol) in
dichloromethane (150 ml). The reaction mixture was allowed to warm
to room temperature for 1 hour. The mixture was concentrated and
the residue was purified by chromatography (heptanes/acetone 1/2)
to afford
[6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-methanol
(4.4 g, 71%) as colorless oil. .sup.1H NMR (400 MHz, CHLOROFORM-d)
.delta. ppm 1.79 (d, J=11.62 Hz, 2H) 2.07-2.25 (m, 2H) 2.59-2.71
(m, 1H) 2.82-2.95 (m, 1H) 3.42 (s, 2H) 3.91 (s, 2H) 4.33 (d, J=9.60
Hz, 1H) 7.30 (d, J=8.72 Hz, 2H) 7.42 (d, J=8.59 Hz, 2H).
Step 5. Synthesis of
6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]octane-4-carbaldehyde
##STR00098##
[0314] To a stirred solution of
[6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-methanol
(2.47 g, 7.45 mmol) in dichloromethane (100 ml) at -78.degree. C.,
was added Dess-Martin periodinane (3.32 g, 7.82 mmol). The reaction
mixture was allowed to warm to 0.degree. C. for 30 minutes and then
room temperature for 60 minutes. Most of the solvent was removed by
evaporation under vacuum at the room temperature and the residue
was purified by flash chromatography (heptanes/acetone 1/2) to
afford
6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]octane-4-carbaldehyde
(2.0 g, 81%) as colorless oil. .sup.1H NMR (400 MHz, CHLOROFORM-d)
.delta. ppm 1.98-2.21 (m, 3H) 2.34-2.44 (m, 1H) 2.69-2.80 (m, 1H)
2.82-2.92 (m, 1H) 3.92-4.08 (m, 2H) 4.21-4.32 (m, 1H) 7.25 (d,
J=8.60 Hz, 2H) 7.34 (d, J=8.60 Hz, 2H) 9.44 (s, 1H).
Step 6. Synthesis of
6-bromo-1-(4-chloro-phenyl)-4-(2-methoxy-vinyl)-2-oxa-bicyclo[2.2.2]octan-
e
##STR00099##
[0316] To a solution of triphenylphosphine methoxymethane chloride
(3.51 g, 10.24 mmol) in tetrahydrofuran (20 ml) under nitrogen at
room temperature, was added potassium t-butoxide (1.149 g, 10.24
mmol). The reddish solution was stirred at room temperature for 90
minutes. Then a solution of
6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]octane-4-carbaldehyde
(2.25 g, 6.83 mmol) in tetrahydrofuran (6 ml) was added to the
reaction mixture. The reaction mixture was stirred at room
temperature for 10 minutes and then quenched with water (2 ml). The
mixture was partitioned between brine (15 ml) and ethyl acetate (60
ml). The aqueous layer was extracted with ethyl acetate (2.times.30
ml) and the combined organic extracts were washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated. The residue was purified
by flash chromatography (heptanes/acetone 2/1) to afford the cis
and trans mixtures of
6-bromo-1-(4-chloro-phenyl)-4-(2-methoxy-vinyl)-2-oxa-bicyclo[2.2.2]octan-
e (2.32 g, 95%) as colorless oil. .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 1.74-2.99 (m, 6H) 3.54 (3.89 (m, 4H)
3.91-6.36 (m, 5H) 7.28-7.43 (m, 4H).
Step 7. Synthesis of
[6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-acetic
acid methyl ester
##STR00100##
[0318] To a stirred solution of the cis and trans mixtures of
6-bromo-1-(4-chloro-phenyl)-4-(2-methoxy-vinyl)-2-oxa-bicyclo[2.2.2]octan-
e (1.7 g, 4.75 mmol) in dichloromethane (12 ml) and tetrahydrofuran
(25 ml) at room temperature, was portionally added pyridinium
chlorochromate (3.07 g, 14.26 mmol). The reaction mixture was
stirred at room temperature for 2 hours. More pyridinium
chlorochromate (2.05 g, 9.51 mmol) was added, and the mixture was
stirred for another 60 hours at room temperature. The mixture was
filtrated and the filtrate was concentrated, and diluted with ethyl
acetate (100 ml). The organic phase was washed with water, brine
and dried over Na.sub.2SO.sub.4, and concentrated. The residue was
purified by flash chromatography (heptanes/acetone 2/1) to afford
[6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-acetic
acid methyl ester (1.08 g, 61%) as colorless oil. .sup.1H NMR (400
MHz, CHLOROFORM-d) .delta. ppm 1.69-1.79 (m, 1H) 1.87-1.98 (m, 1H)
2.01-2.11 (m, 1H) 2.14 (s, 2H) 2.23-2.30 (m, 1H) 2.52-2.63 (m, 1H)
2.75-2.87 (m, 1H) 3.62 (s, 3H) 3.84 (dd, J=10.11, 2.65 Hz, 2H)
4.19-4.29 (m, 1H) 7.23 (d, J=8.80 Hz, 2H) 7.33 (d, J=8.80 Hz,
2H).
Step 8. Synthesis of
[1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-acetic acid
methyl ester
##STR00101##
[0320] To a stirred solution of
[6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-acetic
acid methyl ester (0.75 g, 2.007 mmol) and nickel (II) chloride
(0.182 g, 1.405 mmol) in methanol (30 ml), ethyl acetate (15 ml),
tetrahydrofuran (15 ml) and chlorobenzene (2 ml) at -30.degree. C.,
was added carefully sodium borohydride (0.228 g, 6.02 mmol). The
reaction mixture was allowed to warm to room temperature for 40
minutes. The solvent was removed under vacuum at room temperature
and then water (20 ml) and EtOAc (100 ml) were added. The organic
phase was separated and washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
flash chromatography (heptanes/acetone 2/1) to afford
[1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-acetic acid
methyl ester (0.39 g, 66%) as white solid. .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 1.65-1.81 (m, 4H) 1.96 (t, J=8.00 Hz, 4H)
2.11 (s, 2H) 3.62 (s, 3H) 3.87 (s, 2H) 7.20 (d, J=8.60 Hz, 2H) 7.25
(d, J=8.60 Hz, 2H).
Step 9. Synthesis of
{1-[4'-(5-tert-butyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bic-
yclo[2.2.2]oct-4-yl}-acetic acid methyl ester
##STR00102##
[0322] To a solution of
[1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-acetic acid
methyl ester (0.29 g, 0.984 mmol) and
(5-tert-butyl-[1,3,4]oxadiazol-2-yl)-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxa-
borolan-2-yl)-phenyl]-amine (0.338 g, 0.984 mmol) in dioxane (8 ml)
and water (0.8 ml), was added caesium fluoride (0.448 g, 2.95 mmol)
and Pd(amphos)Cl.sub.2 (0.070 g, 0.098 mmol). The mixture was
flushed with nitrogen for three times and heated at 90.degree. C.
for 16 hours. The reaction mixture was filtered, and the solid was
washed with water (10 ml) and ethyl acetate (3 ml) to afford crude
{1-[4'-(5-tert-butyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bic-
yclo[2.2.2]oct-4-yl}-acetic acid methyl ester (0.27 g, 58%) as
white solid. [MS+1] 476.31.
Step 10. Synthesis of
{1-[4'-(5-tert-butyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bic-
yclo[2.2.2]oct-4-yl}-acetic acid
##STR00103##
[0324] A 100 ml round bottom flask was charged with
{1-[4'-(5-tert-butyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bic-
yclo[2.2.2]oct-4-yl}-acetic acid methyl ester (0.27 g, 0.454 mmol),
and lithium hydroxide monohydrate (0.229 g, 5.45 mmol) in
tetrahydrofuran (20 ml), ethanol (10 ml) and water (8 ml) at room
temperature. The mixture was heated at 50.degree. C. for 2 hours.
Most of the organic solvent was evaporated under vacuum and the
mixture was acidified to PH=4 at 0.degree. C.
[0325] The solvents were then removed by lipherlizer. The mixture
was added ethanol (40 ml) and heated to 70.degree. C., and then was
cooled to room temperature. The mixture was filtered and the solid
was washed with ethanol (3 ml) and water (10 ml) to afford
{1-[4'-(5-tert-butyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bic-
yclo[2.2.2]oct-4-yl}-acetic acid (150 mg, 71%) as white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.64-1.89 (m, 6H)
2.00-2.13 (m, 4H) 3.85 (s, 2H) 7.42 (d, J=8.40 Hz, 2H) 7.55 (d,
J=8.50 Hz, 2H) 7.62 (s, 4H) 10.46 (br. s, 1H). LC/MS, [MS+] 461.23,
RT: 2.66 (Condition M).
Example 19
{1-[4'-(5-cyclobutyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bicy-
clo[2.2.2]oct-4-yl}-acetic acid
Step 1. methyl
2-(1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-4-yl)acetate
##STR00104##
[0327] A vial was charged with
[6-bromo-1-(4-chloro-phenyl)-2-oxa-bicyclo[2.2.2]oct-4-yl]-acetic
acid methyl ester (80 mg, 0.214 mmol), pinoco boronate (163 mg,
0.642 mmol), potassium acetate (63.0 mg, 0.642 mmol), Pd(X-phos)
(31.6 mg, 0.043 mmol) X-phos (30.6 mg, 0.064 mmol) and Dioxane (2
ml) at room temperature. The vial was sealed, flushed with nitrogen
for three times and heated at 102.degree. C. for 16 h. The mixture
was cooled down and diluted with dioxane (5 ml) and EtOAc (50 ml),
filtered and the filtrate was concentrated. The residue was added
dioxane (30 ml) and filtered again, and the filtrate was purified
by flash chromatography (heptanes/ethyl acetate 1/1) to afford
methyl
2-(1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-4-yl)acetate (37 mg, 80% purity, 36% yield) as white
solid. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.26 (s,
12H) 1.66-1.80 (m, 4H) 1.89-2.04 (m, 4H) 2.11 (s, 2H) 3.60 (s, 3H)
3.87 (s, 2H) 7.32 (d, J=8.21 Hz, 2H) 7.69 (d, J=8.08 Hz, 2H).
Step 2. Synthesis of methyl
2-(1-(4'-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)--
2-oxabicyclo[2.2.2]octan-4-yl)acetate
##STR00105##
[0329] A vial was charged with methyl
2-(1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-4-yl)acetate (20 mg, 0.052 mmol),
N-(4-bromophenyl)-5-cyclobutyl-1,3,4-oxadiazol-2-amine (30.5 mg,
0.104 mmol), PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2 adduct (6.34 mg,
7.77 .mu.mol), potassium phosphate (21.98 mg, 0.104 mmol),
dimethoxyethane (0.6 ml), methanol (0.2 ml), and water (0.1 ml) at
room temperature. The vial was sealed, flushed with nitrogen for
three times and heated at 80.degree. C. for 16 hours. The mixture
was purified by flash chromatography (heptanes/acetone 1/1, washed
with acetone/ethanol 1/1) to afford methyl
2-(1-(4'-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-4-yl)acetate (40 mg, 40% purity, 65%
yield) as white solid. LC/MS, ESI-MS(+1) m/z 474.19, RT 1.41
(Condition E).
Step 3. Synthesis of
{1-[4'-(5-cyclobutyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2-oxa-bic-
yclo[2.2.2]oct-4-yl}-acetic acid
##STR00106##
[0331] A flask was charged with methyl
2-(1-(4'-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-yl)-
-2-oxabicyclo[2.2.2]octan-4-yl)acetate (0.040 g, 0.034 mmol),
lithium hydroxide monohydrate (0.043 g, 1.040 mmol),
tetrahydrofuran (1 ml), ethanol (0.3 ml) and water (0.3 ml), and
stirred at 50.degree. C. for 2 hours. The mixture was spurified by
HPLC (0.1% NH.sub.4OH, 10-50% acetonitrile/water, 20 minutes run,
retention time .about.7 minutes) to afford
{1-[4'-(5-cyclobutyl-[1,3,4]oxadiazol-2-ylamino)-biphenyl-4-yl]-2--
oxa-bicyclo[2.2.2]oct-4-yl}-acetic acid (6 mg, 39%) as white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.67-1.89 (m, 5H)
1.89-2.00 (m, 1H) 2.00-2.14 (m, 5H) 2.26-2.39 (m, 5H) 3.62-3.73 (m,
1H) 3.86 (s, 2H) 7.43 (d, J=8.40 Hz, 2H) 7.55 (d, J=8.50 Hz, 2H)
7.62 (s, 4H) 10.39-10.59 (br. s, 1H). LC/MS, [MS+] 459.22, RT: 2.61
(Condition M).
Example 20
(1-{4'-[(2-ethyl-4-methyl-oxazole-5-carbonyl)-amino]-biphenyl-4-yl}-2-oxa--
bicyclo[2.2.2]oct-4-yl)-acetic acid
Step 1. Synthesis of methyl
2-(1-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1-biphenyl]-4-yl)-2-o-
xabicyclo[2.2.2]octan-4-yl)acetate
##STR00107##
[0333] A vial was charged with methyl
2-(1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxabicyclo-
[2.2.2]octan-4-yl)acetate (14 mg, 0.036 mmol),
N-(4-bromophenyl)-2-ethyl-4-methyloxazole-5-carboxamide (22.4 mg,
0.072 mmol), PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2 adduct (4.44 mg,
5.44 .mu.mol), potassium phosphate (15.4 mg, 0.072 mmol),
dimethoxyethane (0.6 ml), ethanol (0.2 ml), and water (0.1 ml) at
room temperature. The vial was sealed, flushed with nitrogen for
three times and heated at 80.degree. C. for 16 hours. The mixture
was purified by flash chromatography (heptanes/acetone 1/1, washed
with acetone/ethanol 1/1) to afford methyl
2-(1-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1'-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-4-yl)acetate (25 mg, 40% purity, 56% yield)
as white solid. LC/MS, ESI-MS(+1) m/z 489.1, RT 1.53 (Condition
E).
Step 2. Synthesis of
(1-{4'-[(2-ethyl-4-methyl-oxazole-5-carbonyl)-amino]-biphenyl-4-yl}-2-oxa-
-bicyclo[2.2.2]oct-4-yl)-acetic acid
##STR00108##
[0335] A flask was charged with methyl
2-(1-(4'-(2-ethyl-4-methyloxazole-5-carboxamido)-[1,1'-biphenyl]-4-yl)-2--
oxabicyclo[2.2.2]octan-4-yl)acetate (0.025 g, 0.020 mmol), lithium
hydroxide monohydrate (0.030 g, 0.716 mmol), tetrahydrofuran (1
ml), ethanol (0.3 ml) and water (0.3 ml), and stirred at 50.degree.
C. for 2 hours. The mixture was spurified by HPLC (0.1% NH.sub.4OH,
10-60% acetonitrile/water, 20 minutes run, retention time .about.7
minutes) to afford
(1-{4'-[(2-ethyl-4-methyl-oxazole-5-carbonyl)-amino]-biphenyl-4-yl-
}-2-oxa-bicyclo[2.2.2]oct-4-yl)-acetic acid (4 mg, 41%) as white
solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.31 (t,
J=7.58 Hz, 3H) 1.68-1.90 (m, 6H) 2.02-2.16 (m, 4H) 2.40 (s, 3H)
2.84 (q, J=7.58 Hz, 1H) 3.87 (s, 2H) 7.44 (d, J=8.46 Hz, 2 H) 7.58
(d, J=8.59 Hz, 2H) 7.63 (d, J=8.72 Hz, 2H) 7.83 (d, J=8.72 Hz, 2H)
10.13 (s, 1H). LC/MS, [MS+] 474.22, RT: 2.65 (Condition M).
Example 21
(1-{4-[5-(5-cyclobutyl-[1,3,4]oxadiazol-2-ylamino)-pyridin-2-yl]-phenyl}-2-
-oxa-bicyclo[2.2.2]oct-4-yl)-acetic acid
Step 1. Synthesis of 2-(4-chlorophenyl)-5-nitropyridine
##STR00109##
[0337] A flask was charged with 2-bromo-5-nitropyridine (4 g, 19.71
mmol), 4-chlorophenylboronic acid (4.54 g, 27.6 mmol), Pd(PPh3)4
(1.139 g, 0.985 mmol), potassium carbonate (8.16 g, 59.1 mmol),
dioxane (40 ml) and water (20 ml) at room temperature. The flask
was sealed, flushed with nitrogen for three times and heated at
100.degree. C. for 3 hours. The reaction mixture was diluted with
EtOAc (100 ml) and filtered. The filtrate was concentrated. The
residue was washed with water by filtration. The afforded solid was
dissolved in EtOAc (200 ml) and washed with saturated aqueous
sodium bicarbonate solution and brine, dried over Na.sub.2SO.sub.4,
concentrated, and dried under vacuum to afford crude
2-(4-chlorophenyl)-5-nitropyridine (4.8 g, 75% purity, 78% yield)
as yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 7.27-7.36
(m, 4H) 7.51-7.61 (m, 3H).
Step 2. Synthesis of
5-nitro-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pyridine
##STR00110##
[0339] A flask was charged with 2-(4-chlorophenyl)-5-nitropyridine
(4.48 g, 14.26 mmol), pinoco boronate (14.48 g, 57.0 mmol),
potassium acetate (5.59 g, 57.0 mmol), Pd-(Xphos) (1.404 g, 1.900
mmol), X-phos (1.359 g, 2.85 mmol) and dioxane (50 ml) at room
temperature. The flask was sealed, flushed with nitrogen for three
times and heated at 115.degree. C. for 72 hours. The mixture was
cooled down to room temperature and diluted with dioxane (50 ml)
and EtOAc (100 ml), filtered and the filtrate was concentrated. The
residue was added dioxane (30 ml) and filtered, the filtrate was
purified by flash chromatography (heptanes/acetone 1/2) to afford
5-nitro-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)p-
yridine (3.9 g, 84%) as yellow solid. .sup.1H NMR (400 MHz,
CHLOROFORM-d) ppm 1.37 (s, 9H) 7.43-8.15 (m, 7H).
Step 3. diethyl
4'-(5-nitropyridin-2-yl)-5,6-dihydro-[1,1'-biphenyl]-4,4(3H)-dicarboxylat-
e
##STR00111##
[0341] A flask was charged with
5-nitro-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pyridine
(3.9 g, 10.73 mmol) and potassium carbonate (6.77 g, 48.3 mmol) in
dioxane (60 ml) and Water (25 ml), and stirred at room temperature
for 10 minutes under nitrogen. Then Pd(PPh.sub.3).sub.4 (1.240 g,
1.073 mmol) and diethyl
4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1,1-dicarboxylate
(4.22 g, 11.27 mmol) were added to the reaction mixture. The flask
was sealed, flushed with nitrogen for three times and heated at
100.degree. C. for 1 hour. The organic solvent was evaporated under
vacuum and EtOAc (3.times.100 ml) was used to extract and the
combined organic solvent was washed by saturated aqueous sodium
bicarbonate solution, brine, and dried over Na.sub.2SO.sub.4, then
concentrated. The residue was purified by flash chromatography
(heptanes/acetone 1/2) to afford diethyl
4'-(5-nitropyridin-2-yl)-5,6-dihydro-[1,1'-biphenyl]-4,4(3H)-dicarboxylat-
e (2.8 g, 61%) as yellow solid. .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 1.25 (t, J=7.20 Hz, 6H) 2.30 (dt,
J=6.28, 3.24 Hz, 2H) 2.53 (dd, J=4.67, 1.77 Hz, 2H) 2.76 (dd,
J=3.98, 2.08 Hz, 2H) 4.20 (q, J=7.20 Hz, 4H) 6.19 (s, 1H) 7.06-8.88
(m, 7H).
Step 4.
(1-(4-(5-aminopyridin-2-yl)phenyl)-6-bromo-2-oxabicyclo[2.2.2]octa-
n-4-yl)methanol
##STR00112##
[0343] An oven-dried flask under nitrogen charged with diethyl
4'-(5-nitropyridin-2-yl)-5,6-dihydro-[1,1'-biphenyl]-4,4(3H)-dicarboxylat-
e (1.3 g, 3.06 mmol) in tetrahydrofuran (60 ml), was added lithium
alumina hydride (15.31 ml, 15.31 mmol) in one portion at 0.degree.
C. The mixture was allowed to warm to room temperature and stirred
for 2 hours. The reaction was quenched with saturated aqueous
sodium chloride solution at 0.degree. C. The mixture was extreacted
with EtOAc (2.times.150 ml), and the combined organic phase was
dried over Na.sub.2SO.sub.4, and concentrated to afford the crude
intermediate, which was carried to the next step without further
purification.
[0344] To a stirred solution of crude intermediate from last step
in tetrahydrofuran (100 ml) at -78.degree. C., was added
N-bromosuccinimide (0.534 g, 3.00 mmol) in dichloromethane (50 ml).
The reaction mixture was stirred at room temperature for overnight.
The mixture was concentrated and the residue was purified by flash
chromatography (heptanes/acetone 1/2) to afford
(1-(4-(5-aminopyridin-2-yl)phenyl)-6-bromo-2-oxabicyclo[2.2.2]octan-4-yl)-
methanol (0.24 g, 21%) as yellow solid. .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 2.03-2.10 (m, 2H) 2.13-2.19 (m, 2H)
2.41-2.53 (m, 2H) 3.45-3.59 (m, 4H) 6.07-6.24 (m, 1H) 7.48 (d,
J=8.59 Hz, 2H) 7.77 (d, J=8.72 Hz, 1H) 7.84 (d, J=8.46 Hz, 2H) 8.13
(dd, J=8.72, 2.53 Hz, 1H) 8.74 (d, J=2.27 Hz, 1H). LC/MS, not
ionized; RT: 0.84; (Condition E).
Step 5. Synthesis of
(1-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-4-yl)methanol
##STR00113##
[0346] To a stirred solution of
(1-(4-(5-aminopyridin-2-yl)phenyl)-6-bromo-2-oxabicyclo[2.2.2]octan-4-yl)-
methanol (240 mg, 0.617 mmol) in toluene (8 ml) and tetrahydrofuran
(8 ml) at 40.degree. C. in a microwave vial under nitrogen, was
added the mixture of AIBN (40.5 mg, 0.247 mmol) and Bu.sub.3SnH
(0.980 ml, 3.70 mmol) in toluene at 40.degree. C. The reaction
mixture was heated at 100.degree. C. for 3 hours. Most of solvent
was removed by evaporation under vacuum, and the residue was
purified by chromotography (heptanes/acetone 1/2) to afford
(1-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-4-yl)methanol
(60 mg, 31%) as yellow solid. LC/MS, [MS+1] 311.1, RT: 0.80;
(Condition E).
Step 6.
(1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)ph-
enyl)-2-oxabicyclo[2.2.2]octan-4-yl)methanol
##STR00114##
[0348] To a solution of
(1-(4-(5-aminopyridin-2-yl)phenyl)-2-oxabicyclo[2.2.2]octan-4-yl)methanol
(50 mg, 0.161 mmol) in dichloromethane (2 ml) and tetrahydrofuran
(1 ml) at room temperature, was added
1,1'-thiocarbonyldipyridin-2(1H)-one (41.2 mg, 0.177 mmol). The
mixture was stirred at room temperature for 1 hour.
[0349] Cyclobutanecarbohydrazide (0.028 g, 0.242 mmol) in
dichloromethane (4 ml) was added to the reaction mixture. The
mixture was stirred at room temperature for 1 hour, and EDC.HCl
(0.093 g, 0.483 mmol) was added. The mixture was stirred at room
temperature for 16 hours. The reaction mixture was diluted with
dichloromethane (30 ml) and was filtered to give
1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-2--
oxabicyclo[2.2.2]octan-4-yl)methanol (30 mg, 43%) as white solid.
.sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm 1.56-2.53 (m,
14H) 3.63-3.78 (m, 2H) 3.94 (s, 2H) 4.10 (d, J=7.07 Hz, 1H)
7.42-7.57 (m, 2H) 7.78-7.92 (m, 3H) 8.15 (dd, J=8.72, 2.78 Hz, 1H)
8.71 (dd, J=2.72, 0.57 Hz, 1H). LC/MS, [MS+1] 433.1, RT: 1.11;
(Condition E).
Step 7.
1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phe-
nyl)-2-oxabicyclo[2.2.2]octane-4-carbaldehyde
##STR00115##
[0351] To a stirred solution of
1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-2--
oxabicyclo[2.2.2]octan-4-yl)methanol (35 mg, 0.081 mmol) in
dichloromethane (1 ml) and DMSO (0.5 ml) at 0.degree. C., was added
diisopropylethylamine (0.057 ml, 0.324 mmol) and Py.SO.sub.3 (25.8
mg, 0.162 mmol). The reaction mixture was stirred at room
temperature for 2 hours. The reaction mixture was purified by flash
chromatography (heptanes/acetone 1/1) to afford
1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-2--
oxabicyclo[2.2.2]octane-4-carbaldehyde (30 mg, 86%) as yellow
solid. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.83-2.56
(m, 14H) 3.56-3.74 (m, 1H) 4.15 (s, 2H) 7.50 (d, J=8.46 Hz, 2H)
7.77 (d, J=8.59 Hz, 1H) 7.94 (d, J=8.46 Hz, 2H) 8.16-8.37 (m, 1H)
8.56-8.72 (m, 1H) 9.55 (s, 1H). LC/MS, [MS+1] 431.1, RT: 1.24;
(Condition E).
Step 8.
5-cyclobutyl-N-(6-(4-(4-(2-methoxyvinyl)-2-oxabicyclo[2.2.2]octan--
1-yl)phenyl)pyridin-3-yl)-1,3,4-oxadiazol-2-amine
##STR00116##
[0353] To a stirred suspension of triphenylphosphine methoxymethane
chloride (28.7 mg, 0.084 mmol) in tetahydrofuran (1 ml) was flushed
with nitrogen for three times and at 0.degree. C., and was added
LiHMDS (0.079 ml, 0.084 mmol). The reddish solution was stirred at
0.degree. C. for 30 minutes, and then at -78.degree. C., a solution
of
1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-2--
oxabicyclo[2.2.2]octane-4-carbaldehyde (30 mg, 0.070 mmol) in
tetrahydrofuran (0.7 ml) was added dropwise. The mixture was
allowed to warm to 0.degree. C. for 30 minutes. The reaction was
quenched with water (2 ml), and the mixture was partitioned between
brine (15 ml) and ethyl acetate (50 ml), aqueous layer was
extracted with ethyl acetate (2.times.50 ml). The combined organic
extracts were washed with brine and dried over Na.sub.2SO.sub.4,
and concentrated. The residue was purified by flash chromatography
(heptanes/acetone 1/1) to afford
5-cyclobutyl-N-(6-(4-(4-(2-methoxyvinyl)-2-oxabicyclo[2.2.2]octan-1-yl)ph-
enyl)pyridin-3-yl)-1,3,4-oxadiazol-2-amine (10 mg, 31%) as yellow
solid. .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm
1.83-2.51 (m, 14H) 3.56 (s, 3H) 3.63-3.76 (m, 1H) 4.00 (d, J=6.80
Hz, 1H) 4.06 (s, 2H) 5.87 (d, J=6.80 Hz, 1H) 7.49 (d, J=8.72 Hz,
2H) 7.78-7.93 (m, 3H) 8.17 (dd, J=24.00, 8.80 Hz, 1H) 8.71 (d,
J=16.80 Hz, 1H). LC/MS, [MS+1] 459.1, RT: 1.44, 1.48; (Condition
E).
Step 9. Synthesis of
(1-{4-[5-(5-cyclobutyl-[1,3,4]oxadiazol-2-ylamino)-pyridin-2-yl]-phenyl}--
2-oxa-bicyclo[2.2.2]oct-4-yl)-acetic acid
##STR00117##
[0355] To a stirred solution of
5-cyclobutyl-N-(6-(4-(4-(2-methoxyvinyl)-2-oxabicyclo[2.2.2]octan-1-yl)ph-
enyl)pyridin-3-yl)-1,3,4-oxadiazol-2-amine (10 mg, 0.022 mmol) in
tetrahydrofuran (1 ml) at room temperature, was added 1N aqueous
HCl solution (0.436 ml, 0.436 mmol). The reaction mixture was
stirred for 1 h at room temperature. Most of solvent was removed by
evaporation under vacuum, and the residue was purified by flash
chromotography to afford
2-(1-(4-(5-((5-cyclobutyl-1,3,4-oxadiazol-2-yl)amino)pyridin-2-yl)phenyl)-
-2-oxabicyclo[2.2.2]octan-4-yl)acetaldehyde intermediate.
[0356] To a stirred solution of the intermediate from last step and
2-methyl-2-butene (0.112 ml, 0.225 mmol) in t-butanol (1 ml), water
(0.3 ml), tetahydrofuran (1.000 ml) cooling with ice-cold bath, was
added NaClO.sub.2 (3.31 mg, 0.029 mmol). The reaction mixture was
stirred for 1 hour at room temperature. The mixture was filtered
and the filtrate was purified by HPLC (0.1% NH.sub.4OH, 15-100%
acetonitrile/water) to afford
(1-{4-[5-(5-cyclobutyl-[1,3,4]oxadiazol-2-ylamino)-pyridin-2-yl]-phenyl}--
2-oxa-bicyclo[2.2.2]oct-4-yl)-acetic acid (5 mg, 43%) as white
solid. .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm
1.26-1.38 (m, 3H) 1.81-1.92 (m, 4H) 1.97-2.08 (m, 3H) 2.10 (s, 2H)
2.11-2.22 (m, 4H) 3.68-3.75 (m, 1H) 4.00 (s, 2H) 7.50 (d, J=8.40
Hz, 2H) 7.80-7.89 (m, 3H) 8.16 (dd, J=9.60, 2.80 Hz, 1H) 8.71 (dd,
J=2.80, 0.60 Hz, 1H). LC/MS, [MS+] 460.21, RT: 2.45 (Condition
M).
Example 22
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-
-7-oxabicyclo[2.2.1]heptan-1-yl)acetic acid
Step 1. Synthesis of
8-(4-bromophenyl)-1,4-dioxaspiro[4.5]decan-8-ol
##STR00118##
[0358] To a flask charged with dry, freshly ground magnesium
turnings (187 mg, 7.7 mmol) and I.sub.2 (81 mg, 0.32 mmol) in THF
(10 mL) was added a solution of 1,4-dibromobenzene (1.81 g, 7.7
mmol) in THF (10 mL) slowly. The resulting mixture was stirred at
rt for 30 min. The resultant pale yellow Grignard solution was
cooled to -78.degree. C. and to it was added a solution of
1,4-dioxaspiro[4.5]decan-8-one (1 g, 6.4 mmol) in THF (10 mL)
slowly dropwise. The reaction was stirred at -78.degree. C. for 20
min before being allowed to stir and warm to rt for 15 hr. The
reaction was quenched with a saturated aqueous NH.sub.4Cl solution
and extracted with methyl tert-butyl ether. The organic phase was
washed with a saturated aqueous NaCl solution, dried over
MgSO.sub.4, filtered and concentrated by rotary evaporation in
vacuo to afford 2.2 g of crude orange oil. Purification of the
crude material by column chromatography (50 g silica gel, 0-50%
EtOAc:Hept, monitor for 225 nm) afforded 0.99 g of the title
compound as a white solid.
[0359] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. ppm 1.47-1.58
(m, 2H), 1.58-1.70 (m, 2H), 1.82-2.02 (m, 4H), 3.88 (s, 4H), 4.99
(s, 1H), 7.40 (d, J=8.6 Hz, 2H), 7.49 (d, J=8.6 Hz, 2H). ESI-MS
m/z: not ionized [M+H].sup.+, retention time 1.29 min (condition
E).
Step 2. Synthesis of 4-(4-bromophenyl)-4-hydroxycyclohexanone
##STR00119##
[0361] To a mixture of
8-(4-bromophenyl)-1,4-dioxaspiro[4.5]decan-8-ol (0.99 g, 3.2 mmol)
in acetone (10 mL) and water (5 mL) was added Ts-OH (12 mg, 0.063
mmol) and the reaction was let stir in at 75.degree. C. oil bath
for 1 hr. The resulting reaction mixture was cooled to rt and the
acetone was removed by rotary evaporation in vacuo. The resulting
aqueous mixture was extracted with ethyl acetate. The organic phase
was dried over MgSO.sub.4, filtered and concentrated by rotary
evaporation in vacuo to afford 0.9 g of a crude pale yellow solid
which was used without further purification. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. ppm 1.84-1.98 (m, 2H), 2.06-2.29 (m, 4H),
2.76 (td, J=13.8, 6.3 Hz, 2H), 5.47 (s, 1H), 7.51 (s, 4H). ESI-MS
m/z: not ionized [M+H].sup.+, retention time 1.18 min (condition
E).
Step 3. Synthesis of methyl
2-(4-(4-bromophenyl)-4-hydroxycyclohexylidene)acetate
##STR00120##
[0363] To a solution of NaH (60% dispersion in mineral oil, 70 mg,
1.7 mmol) in methanol (6.7 mL) was added trimethylphosphonoacetate
(0.23 mL, 1.6 mmol) and the reaction was let stir at rt for 30 min.
To the reaction was added 4-(4-bromophenyl)-4-hydroxycyclohexanone
(360 mg, 1.3 mmol) and the reaction was let stir at rt for 20 h.
The resulting reaction mixture was quenched with saturated aqueous
ammonium chloride solution and extracted with ethyl acetate. The
organic phase was dried over MgSO.sub.4, filtered and concentrated
by rotary evaporation in vacuo to afford 530 mg of a crude opaque
oil. Purification of the crude material by column chromatography
(25 g silica gel, 0-60% EtOAc:Hept, monitor 230 nm) afforded 51 mg
(12%) of methyl
2-(4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)acetate as a
colorless oil (ESI-MS m/z: 327.1 [M+H].sup.+, retention time 1.56
min (condition E)) and 275 mg (63%) of the title compound as a
colorless oil (ESI-MS m/z: n/a [M+H].sup.+, retention time 1.42 min
(condition E)). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. ppm
1.71-1.93 (m, 4H), 2.19 (d, J=13.1 Hz, 1H), 2.29-2.44 (m, 1H), 2.66
(td, J=12.6, 4.7 Hz, 1H), 3.56-3.69 (m, 4H), 5.24 (s, 1H), 5.72 (s,
1H), 7.41-7.52 (m, 4H). .sup.13C NMR (100 MHz, DMSO-d.sub.6):
.delta. ppm 166.1, 162.5, 148.7, 130.6, 127.1, 119.3, 112.6, 70.8,
50.6, 38.7, 32.4, 24.5.
Step 4. Synthesis of methyl
2-(4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)acetate
##STR00121##
[0365] To a solution of methyl
2-(4-(4-bromophenyl)-4-hydroxycyclohexylidene)acetate (190 mg, 0.58
mmol) in acetonitrile (7 mL) was added Cs.sub.2CO.sub.3 (381 mg,
1.17 mmol) and the reaction was let stir in a 50.degree. C. oil
bath for 24 h. The resulting reaction mixture was diluted with
ethyl acetate and washed with water, followed by a saturated
aqueous NaCl solution. The organic phase was dried over MgSO.sub.4,
filtered and concentrated by rotary evaporation in vacuo to afford
188 mg of crude orange film. The crude material was purified by
column chromatography (24 g silica gel, 0-40% EtOAc:Hept, monitor
230 nm) to afford 64 mg (34%) of the title compound as a colorless
film that solidified upon standing. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. ppm 1.56-1.81 (m, 4H), 1.85-1.97 (m, 2H),
2.14 (td, J=9.9, 3.9 Hz, 2H), 2.92 (s, 2H), 3.63 (s, 3H), 7.32 (d,
J=8.6 Hz, 2H), 7.53 (d, J=8.6 Hz, 2H). .sup.13C NMR (100 MHz,
DMSO-d.sub.6): .delta. ppm 170.3, 142.0, 130.9, 127.1, 119.8, 86.0,
83.2, 51.3, 37.6, 35.4 (additional signal likely hidden under DMSO
solvent signal). ESI-MS m/z: 327.0 [M+H].sup.+, retention time 1.55
min (condition E).
Step 5. Synthesis of methyl
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)acetate
##STR00122##
[0367] To a flask charged with methyl
2-(4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)acetate (106
mg, 0.33 mmol),
5-(tert-butyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)p-
henyl)-1,3,4-oxadiazol-2-amine (117 mg, 0.34 mmol),
Pd(amphos)Cl.sub.2 (23 mg, 0.033 mmol) and CsF (150 mg, 0.98 mmol)
under N.sub.2 was added 1,4-dioxane (2.9 mL) and water (0.33 mL).
The mixture was sparged with N.sub.2 for 10 min and set to stir in
a 90.degree. C. oil bath for 2 hr. The reaction mixture was diluted
with ethyl acetate, washed with a saturated aqueous NaHCO.sub.3
solution, then water, followed by a saturated aqueous NaCl
solution. The organic phase was dried over MgSO.sub.4, filtered and
concentrated by rotary evaporation in vacuo to afford 210 mg crude
orange solid. The crude material was triturated with warm
acetonitrile to afford 113 mg of the title compound as a white
solid which was used without further purification. .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 1.36 (s, 9H), 1.66-1.83 (m, 4H),
1.88-2.00 (m, 2H), 2.10-2.21 (m, 2H), 2.94 (s, 2H), 3.64 (s, 3H),
7.42 (d, J=8.6 Hz, 2H), 7.55-7.71 (m, 6H), 10.52 (s, 1H). ESI-MS
m/z: 462.1 [M+H].sup.+, retention time 1.51 min (condition E).
Step 6. Synthesis of
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl-
)-7-oxabicyclo[2.2.1]heptan-1-yl)acetic acid
##STR00123##
[0369] To a solution of methyl
2-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)acetate (110 mg, 0.24 mmol) in
THF (4 mL) and Methanol (1 mL) was added 1N NaOH (0.72 mL, 0.72
mmol) and the reaction was let stir at rt for 2 hr. The resulting
yellow reaction mixture was partially concentrated by rotary
evaporation in vacuo (rt water bath). The yellow solution was
diluted with MeOH, filtered and purified on preparative HPLC
(10-50% MeCN:5 mM NH4OH) to afford 52 mg (49%) of the title
compound as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. ppm 1.36 (s, 9H), 1.63-1.82 (m, 4H), 1.90-2.02 (m, 2H),
2.08-2.21 (m, 2H), 2.79 (s, 2H), 7.42 (d, J=8.3 Hz, 2H), 7.56-7.70
(m, 6H), 10.51 (br. s., 1H). ESI-MS m/z: 448.1 [M+H].sup.+,
retention time 1.08 min (condition E). HR/MS [M+H].sup.+. found
448.2236, calc. 448.2218. RT: 2.27 (Condition L).
Example 23
3-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1-biphenyl]-4-yl)-
-7-oxabicyclo[2.2.1]heptan-1-yl)propanoic acid
Step 1. Synthesis of 4-methylenecyclohexanone
##STR00124##
[0371] To a solution of methyl triphenylphosphonium bromide (13.7
g, 38 mmol) in 1,4-dioxane (60 mL) was added potassium
tert-butoxide (4.3 g, 38 mmol). The resulting yellow mixture was
cooled to 0.degree. C. in an ice-water bath and
1,4-dioxaspiro[4.5]decan-8-one (5 g, 32 mmol) was added slowly over
15 min as a solution in 1,4-dioxane (15 mL) via dropping funnel.
The reaction was let stir and warm to rt over 1 h. The resulting
brown-yellow reaction mixture was partitioned between methyl
tert-butyl ether and saturated aqueous NH.sub.4Cl solution. The
aqueous phase was extracted with methyl tert-butyl ether. The
combined organic phases were washed with a saturated aqueous NaCl
solution, dried over MgSO.sub.4, filtered and concentrated by
rotary evaporation in vacuo to .about.20 mL. The resultant beige
suspension was diluted with heptane and filtered. The filtrated was
concentrated by rotary evaporation in vacuo to afford .about.7 g of
crude 8-methylene-1,4-dioxaspiro[4.5]decane as a yellow liquid
containing residual heptane.
[0372] The crude liquid was taken up in acetone (15 mL) and water
(15 mL) and to this was added toluenesulfonic acid (5 mg). The
reaction was let stir at 50.degree. C. for 8 h. The resulting
yellow reaction mixture was concentrated to .about.20 mL by rotary
evaporation in vacuo and partitioned between methyl tert-butyl
ether and saturated aqueous NH.sub.4Cl solution. The aqueous phase
was extracted with methyl tert-butyl ether. The combined organic
phases were washed with a saturated aqueous NaCl solution, dried
over MgSO.sub.4, filtered and concentrated to -5 mL by rotary
evaporation in vacuo to afford .about.5 g of yellow liquid that was
used without further purification.
Step 2. Synthesis of 1-(4-bromophenyl)-4-methylenecyclohexanol
##STR00125##
[0374] To a flask charged with dry, freshly ground magnesium
turnings (1.1 g, 45 mmol) and iodine (25 mg, 0.1 mmol) in THF (50
mL) was added a solution of 1,4-dibromobenzene (10.6 g, 45.0 mmol)
in THF (30 mL) slowly by dropping funnel over 30 min. The resulting
mixture was stirred at rt for 1 hr. The resultant grey Grignard
solution was cooled to -78.degree. C. and to it was added a
solution of 4-methylenecyclohexanone (3.3 g, 30 mmol) in THF (20
mL) slowly dropwise over 20 min. The reaction was stirred at
-78.degree. C. for 30 min before being allowed to stir and warm to
rt for 3 h. The resulting reaction mixture was quenched with a
saturated aqueous NH.sub.4Cl solution and extracted with methyl
tert-butyl ether. The combined organic phases were washed with a
saturated aqueous NaCl solution, dried over MgSO.sub.4, filtered
and concentrated by rotary evaporation in vacuo to afford a crude
yellow oil. Purification of the crude material by column
chromatography (80 g silica gel, 0-50% EtOAc:Hept, monitor for 225
nm) afforded 2.94 g of a colorless oil that was used without
further purification. ESI-MS m/z: not ionized [M+H].sup.+,
retention time 1.50 min (condition E).
Step 3. Synthesis of
1-(4-bromophenyl)-4-(iodomethyl)-7-oxabicyclo[2.2.1]heptanes
##STR00126##
[0376] To a suspension of Na.sub.2CO.sub.3 (0.63 g, 6.0 mmol) in a
solution of 1-(4-bromophenyl)-4-methylenecyclohexanol (1.06 g, 4.0
mmol) in acetonitrile (20 mL) was added iodine (2.0 g, 7.9 mmol)
portion-wise over 20 min and the reaction was let stir at rt for 4
h. The resulting dark red reaction was diluted with EtOAc, washed
sequentially with a saturate aqueous Na.sub.2S.sub.2O.sub.3
solution, water followed by a saturated aqueous NaCl solution. The
organic phase was dried over MgSO.sub.4, filtered and concentrated
by rotary evaporation in vacuo to afford a crude yellow residue.
Purification of the crude material by column chromatography (12 g
silica gel, 0-30% EtOAc:Hept, monitor 225 nm) afforded 1 g white
solid that was used without further purification. .sup.1H NMR (400
MHz, DMSO-d.sub.6): .delta. ppm 1.62-1.86 (m, 4H), 1.87-1.98 (m,
2H), 2.17-2.29 (m, 2H), 3.69 (s, 2H), 7.32 (d, J=8.6 Hz, 2H), 7.54
(d, J=8.6 Hz, 2H). ESI-MS m/z: not ionized [M+H].sup.+, retention
time 1.73 min (condition E).
Step 4. Synthesis of dimethyl
2-((4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)methyl)malonate
##STR00127##
[0378] To a solution of dimethyl malonate (0.47 mL, 4.1 mmol) in
anhydrous dimethyl acetamide (5.6 mL) cooled in an ice-water bath
was added NaH (60% dispersion in mineral oil, 163 mg, 4.1 mmol)
portion-wise and the reaction was let stir at rt for 30 min until
homogeneous. To this was added
1-(4-bromophenyl)-4-(iodomethyl)-7-oxabicyclo[2.2.1]heptane (440
mg, 1.12 mmol) and the reaction was stirred in a microwave reactor
at 150.degree. C. for 45 min. The resulting yellow reaction mixture
was quenched with a saturated aqueous NH.sub.4Cl solution and
extracted with EtOAc. The organic phase was dried over MgSO.sub.4,
filtered and concentrated by rotary evaporation in vacuo to afford
a crude yellow oil. Purification of the crude material by column
chromatography (40 g silica gel, 0-30% EtOAc:Hept, monitor 225 nm)
afforded 108 mg (28%) of methyl
3-(4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoate
(decaboxylated material) as a clear yellow film and 245 mg (55%) of
the title compound as a clear yellow film. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. ppm 1.54-1.79 (m, 6H), 2.06-2.16 (m, 2H),
2.39 (d, J=7.1 Hz, 2H), 3.66 (s, 6H), 3.73 (t, J=7.1 Hz, 1H), 7.27
(d, J=8.6 Hz, 2H), 7.54 (d, J=8.6 Hz, 2H). ESI-MS m/z: 399.0
[M+H].sup.+, retention time 1.59 min (condition E).
Step 5. Synthesis of methyl
3-(4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoate
##STR00128##
[0380] To a solution of dimethyl
2-((4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)methyl)malonate
(305 mg, 0.77 mmol) in DMSO (3.0 mL) and water (0.03 mL) was added
lithium chloride (98 mg, 2.3 mmol) and the reaction was stirred in
a microwave reactor at 180.degree. C. for 20 min. The resulting
yellow reaction mixture was diluted with water and extracted with
EtOAc. The organic phase was dried over MgSO.sub.4, filtered and
concentrated by rotary evaporation in vacuo to afford 289 mg of a
crude clear yellow film. Purification of the crude material by
column chromatography (24 g silica gel, 0-30% EtOAc:Hept, monitor
230 nm) afforded 194 mg colorless film that solidified upon
standing and which was used without further purification. .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. ppm 1.54-1.67 (m, 4H),
1.68-1.79 (m, 2H), 2.04-2.16 (m, 4H), 2.42-2.48 (m, 2H), 3.61 (s,
3H), 7.31 (d, J=8.6 Hz, 2H), 7.53 (d, J=8.6 Hz, 2H). ESI-MS m/z:
341.1 [M+H].sup.+, retention time 1.59 min (condition E).
Step 6. Synthesis of methyl
3-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoate
##STR00129##
[0382] To a flask charged with methyl
3-(4-(4-bromophenyl)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoate (103
mg, 0.30 mmol),
5-(tert-butyl)-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)--
1,3,4-oxadiazol-2-amine (109 mg, 0.32 mmol), Pd(amphos)Cl.sub.2 (21
mg, 0.030 mmol) and CsF (138 mg, 0.91 mmol) under N.sub.2 was added
1,4-dioxane (2.7 mL) and water (0.30 mL). The mixture was sparged
with N.sub.2 for 10 min and stirred in a 90.degree. C. oil bath for
2 h. The resulting reaction mixture was diluted with
dichloromethane and washed with a saturated aqueous NaHCO.sub.3
solution. The organic layer was warmed and filtered (no product
evident in filtercake). The filtrate was concentrated to afford 200
mg of crude orange solid. The crude material was washed with
triturated with acetonitrile and filtered to afford 106 mg of pale
orange solid which was used without further purification. .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. ppm 1.36 (s, 9H), 1.56-1.83
(m, 6H), 2.04-2.20 (m, 4H), 3.62 (s, 3H), 7.42 (d, J=8.6 Hz, 2H),
7.55-7.69 (m, 6H), 10.51 (s, 1H) (additional 2H signal likely
hidden under DMSO solvent peak). ESI-MS m/z: 476.0 [M+H].sup.+,
retention time 1.55 min (condition E).
Step 7. Synthesis of
3-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoic acid
##STR00130##
[0384] To a solution of methyl
3-(4-(4'-((5-(tert-butyl)-1,3,4-oxadiazol-2-yl)amino)-[1,1'-biphenyl]-4-y-
l)-7-oxabicyclo[2.2.1]heptan-1-yl)propanoate (100 mg, 0.21 mmol) in
THF (3.4 mL) and methanol (0.85 mL) was added 1N NaOH solution
(0.63 mL, 0.63 mmol) and the reaction was let stir at rt for 2 h.
The resulting yellow reaction mixture was partially concentrated by
rotary revaporation (rt water bath) to remove THF. The resulting
yellow mixture was diluted with methanol, filtered and purified on
preparative HPLC (10-50% acetonitrile:5 mM NH4OH) and lyophilized
to afford 83 mg (86%) of the title compound as a white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. ppm 1.36 (s, 9H),
1.56-1.82 (m, 6H), 2.01-2.10 (m, 2H), 2.10-2.19 (m, 2H), 2.30-2.39
(m, 2H), 7.43 (d, J=8.6 Hz, 2H), 7.60 (d, J=8.6 Hz, 2H), 7.62-7.67
(m, 4H) 10.51 (br. s., 1H). .sup.13C NMR (100 MHz, DMSO-d.sub.6):
.delta. ppm 174.5, 165.9, 159.5, 141.7, 138.3, 138.2, 133.1, 127.1,
125.8, 125.4, 117.1, 86.1, 85.5, 37.9, 34.9, 31.7, 30.2, 30.1,
27.8. ESI-MS m/z: 462.1 [M+H].sup.+, retention time 1.13 min
(condition E). HR/MS [M+H].sup.+. found 462.2371, calc. 462.2393.
RT: 1.96 (Condition L).
Biological Assays
[0385] The activity of compounds according to the invention can be
assessed by the following inhibition assay.
DGAT1 Inhibition Assay
[0386] The enzyme preparation used in this assay is a membrane
preparation from Sf9 cells overexpressing human (His).sub.6DGAT1.
During all steps samples were chilled to 4.degree. C. Sf9 cells
expressing human (His).sub.6DGAT1 were thawed at room temperature
and re-suspended at a 10:1 ratio (mL buffer/g of cells) in 50 mM
HEPES, lx Complete Protease Inhibitor, pH 7.5. The re-suspended
pellet was homogenized for 1 min using a Brinkman PT 10/35
homogenizer with a 20 mm generator. Cells were lysed using Avestin
Emulsiflex (chilled to 4.degree. C.) at 10000-15000 psi. Lysate was
centrifuged at 100,000.times.g for 1 h at 4.degree. C. Supernatant
was removed and pellets were re-suspended in 50 mM HEPES, 1.times.
Complete Protease Inhibitor, pH 7.5 at 1/6 the volume of
supernatant. Re-suspended pellets were pooled and homogenized with
10 strokes of a Glas-Col motor driven teflon pestle on setting 70.
The protein concentration of the membrane preparation was
quantified using BCA protein assay with 1% SDS. The membrane
preparation was aliquoted, frozen on dry ice, and stored at
-80.degree. C.
[0387] For 50 mL, 25 mL of 0.2 M HEPES stock buffer, 0.5 mL of 1 M
MgCl.sub.2 (5 mM final concentration), and 24.5 mL of milli-Q
H.sub.2O are added to the 55 mL Wheaton Potter-Elvehjem
homogenizer. Enzyme preparation (0.1 mL) is added to buffer and the
mixture is homogenized with 5 strokes on ice using the Glas-Col
variable speed homogenizer system on setting 70.
[0388] For 50 mL, 0.5 mL 10 mM diolein is added to 9.5 mL of EtOH
in a 50 mL Falcon screw cap conical centrifuge tube. Five mL of 10
mM sodium acetate pH 4.5 is added followed by 0.5 mL of 10 mM
oleoyl-CoA. Finally, the remaining 4.5 mL of 10 mM sodium acetate
pH 4.5 is added followed by 30 mL of milli-Q H20. The solution
should be gently agitated by hand to induce mixing. The final
concentrations of EtOH and sodium acetate are 20% and 2 mM,
respectively.
[0389] Dry compounds are dissolved in the appropriate volume of
DMSO to a final concentration of 10 mM. A 10-point, 3-fold dose
response is used to evaluate compound potency. All dilutions are
performed in DMSO in a Greiner 384-well microplate. [0390] 1. 2
.mu.L of compound in DMSO is added to the appropriate wells. 2
.mu.L of DMSO is added to 100% activity and 100% inhibition
controls. [0391] 2. 25 .mu.L of enzyme mix is added to all wells
and plate(s) are incubated for 10 min at RT. [0392] 3. 10 .mu.L of
20% acetic acid quench is added to 100% inhibition control wells.
Plate(s) are vortexed using Troemner multi-tube vortexer (setting 7
for 10 sec). [0393] 4. 25 .mu.L of substrate mix is added to all
wells. Plate(s) are vortexed using Troemner multi-tube vortexer
(setting 7 for 10 sec). Plate(s) are incubated for 30 min at RT.
[0394] 5. 10 .mu.L of 20% acetic acid quench is added to all wells.
Plate(s) are vortexed using Troemner multi-tube vortexer (setting 7
for 10 sec). [0395] 6. 50 .mu.L of 1-butanol w/glyceryl
tripalmitoleate internal standard is added to all wells. [0396] 7.
Plate(s) are sealed with super pierce strong plate sealer using the
thermo-sealer. [0397] 8. Plate(s) are vortexed using Troemner
multi-tube vortexer (setting 10 for 5 min). [0398] 9. Plate(s) are
centrifuged at 162.times.g (1000 rpm for GH-3.8 rotor) for 5 min
using Beckman GS-6R tabletop centrifuge. Samples were analyzed by
LC/MS/MS using a Waters 1525.mu. LC and Quattro Micro API MS. Where
indicated, tripalmitolein was used as an internal standard to
control for instrument variation.
HPLC Conditions:
[0399] Column: Thermo Betabasic 4, 2.1.times.20 mm
[0400] Solvent: 10 mM Ammonium Formate, 0.1% Formic Acid, 2% water,
98% Methanol
[0401] Isocratic run 0.5 ml per minute
[0402] Run time 1 minute
[0403] Data is converted to % inhibition prior to curve fitting
using the following equation:
% Inhibition = ( response compound - response 100 % inhibition
control ) ( response 100 % activity control - response 100 %
inhibition control .times. 100 ##EQU00001##
[0404] Using the method described above, the compounds of the
present invention were shown to possess inhibitory activity with
IC.sub.50 values ranging from 0.001 .mu.M to 100 .mu.M.
Cellular Assay to Measure Activity of DGAT1 Inhibitors in Mammalian
Cells.
[0405] C2C12 cells are an immortal mouse skeletal muscle cell line
showing an 8-fold enrichment for M DGAT1 versus DGAT2. C2C12 cells
were routinely cultured in 150 cm2 flasks with DMEM (25 mM glucose)
containing 10% FBS, 4 mM L-glutamine, 100 U/ml penicillin and 100
.mu.g/ml streptomycin (30 ml per flask), at 37.degree. C., 5% CO2
and 95% humidity. All studies were performed on cells at passage 10
or less.
[0406] C2C12 cells were seeded in 96-well plates in DMEM containing
4.5 mM glucose and 10% FBS, 18 h (at 37.degree. C.) prior to assay
(all wells of the plate are used). Following 18 h incubation,
seeding medium was then replaced with DMEM (5 mM glucose)
containing 250 .mu.M oleate (complexed to BSA) and compounds or
DMSO, for 2 h (at 37.degree. C.). Compounds were added at 1:3
dilution, 11 points and DMSO control, the starting concentration
was usually 40 .mu.M. Each point was dosed in quadruplicate
allowing 2 compounds to be dosed per plate. The medium was removed
at the end of the incubation and 200 .mu.l/well of 1-butanol added.
The 1-butanol contained an internal standard, tripalmitolein (2
.mu.M). The plates were sealed with an adhesive plate sealer and
left at room temperature for at least 30 min, then spun down at
209.times.g for 5 min.
[0407] Butanolic extracts were transferred to 384-well LC-MS plates
(80 .mu.l/well) and the plate heatsealed with a foil plate sealer.
The 384-well plates containing sample were spun down at 209.times.g
for 5 min prior to loading on the LC-MS.
[0408] Samples were analyzed by LC/MS using a Waters 1525.mu. LC
and Quattro Micro API MS.
[0409] Where indicated, tripalmitolein was used as an internal
standard to control for instrument variation. HPLC conditions as
above.
[0410] Table 1 below shows the inhibitory activity (IC.sub.50
values) of representative compounds to human DGAT1.
TABLE-US-00002 TABLE 1 Activities of compounds of the invention in
the DGAT1 assay DGAT1- Exam- DGAT1 C2C12 ple Name IC.sub.50 (.mu.M)
IC.sub.50 (.mu.M) 1 2-(4-(4-(5-(5-tert-butyl-1,3,4- 0.345 0.288
oxadiazol-2-ylamino)pyridin-2-
yl)phenyl)-2-oxabicyclo[2.2.2]octan-1- yl)acetic acid 2
2-(4-(4-(5-((5-cyclobutyl-1,3,4- 0.12 0.4125
oxadiazol-2-yl)amino)pyridin-2-
yl)phenyl)-2-oxabicyclo[2.2.2]octan-1- yl)acetic acid 3
2-(4-(4-(5-((5-cyclobutyl-1,3,4- 0.0535 0.015
thiadiazol-2-yl)amino)pyridin-2-
yl)phenyl)-2-oxabicyclo[2.2.2]octan-1- yl)acetic acid 4
2-(4-(4-(2-(5-cyclobutyl-1,3,4- 14 2.5
thiadiazol-2-ylamino)pyrimidin-5-
yl)phenyl)-2-oxabicyclo[2.2.2]octan-1- yl)acetic acid 5
2-(4-(4-(6-((5-cyclobutyl-1,3,4- 0.375 1.24
thiadiazol-2-yl)amino)pyridin-3-
yl)phenyl)-2-oxabicyclo[2.2.2]octan-1- yl)acetic acid 6
2-(4-(4'-(5-cyclobutyl-1,3,4-thiadiazol- 0.0125 0.002
2-ylamino)biphenyl-4-yl)-2- oxabicyclo[2.2.2]octan-1-yl)acetic acid
7 2-(4-(4'-(5-cyclobutyl-1,3,4-oxadiazol- 0.0902 0.016
2-ylamino)biphenyl-4-yl)-2- oxabicyclo[2.2.2]octan-1-yl)acetic acid
8 2-(4-(4'-(5-tert-butyl-1,3,4-oxadiazol- 0.099 0.009
2-ylamino)biphenyl-4-yl)-2- oxabicyclo[2.2.2]octan-1-yl)acetic acid
9 2-(4-(4'-((5-cyclopropyl-1,3,4- 230 528
oxadiazol-2-yl)amino)-[1,1'-biphenyl]-
4-yl)-2-oxabicyclo[2.2.2]octan-1- yl)acetic acid 10
2-(4-(4'-(2-ethyl-N,4-dimethyloxazole- 0.36 0.056
5-carboxamido)-[1,1'-biphenyl]-4-yl)-
2-oxabicyclo[2.2.2]octan-1-yl)acetic acid 11
2-(4-(4'-(2-Ethyl-4-methyloxazole-5- 0.93 0.056
carboxamido)biphenyl-4-yl)-.2- oxabicyclo[2.2.2]octan-1-yl)acetic
acid 12 2-(4-(4-(5-(2-methyl-5- 0.8775 NA
(trifluoromethyl)oxazole-4- carboxamido)pyridin-2-yl)phenyl)-2-
oxabicyclo[2.2.2]octan-1-yl)acetic acid 13
2-(4-(4-(5-(2-ethyl-4-methyloxazole-5- 0.68 0.098
carboxamido)pyridin-2-yl)phenyl)-2-
oxabicyclo[2.2.2]octan-1-yl)acetic acid 14
2-(4-(4-(5-((5-(tert-butyl)oxazol-2- 0.13 0.108
yl)amino)pyridin-2-yl)phenyl)-2- oxabicyclo[2.2.2]octan-1-yl)acetic
acid 15 2-(4-(4'-((5-(tert-butyl)oxazol-2- 0.044 0.007
yl)amino)-[1,1'-biphenyl]-4-yl)-2-
oxabicyclo[2.2.2]octan-1-yl)acetic acid 16
2-(4-(4'-((5-isobutyl-1,3,4-oxadiazol-2- 0.06 0.038
yl)amino)-[1,1'-biphenyl]-4-yl)-2-
oxabicyclo[2.2.2]octan-1-yl)acetic acid 17
2-(4-(4'-((5-neopentyl-1,3,4- 0.053 0.031
oxadiazol-2-yl)amino)-[1,1'-biphenyl]-
4-yl)-2-oxabicyclo[2.2.2]octan-1- yl)acetic acid 18
2-(1-(4'-(2-ethyl-5-methyloxazole-4- 2.9 0.16
carboxamido)-[1,1'-biphenyl]-4-yl)-2-
oxabicyclo[2.2.2]octan-4-yl)acetic acid 19
2-(1-(4'-((5-cyclobutyl-1,3,4- 0.26 0.006
oxadiazol-2-yl)amino)-[1,1'-biphenyl]-
4-yl)-2-oxabicyclo[2.2.2]octan-4- yl)acetic acid 20
2-(1-(4'-((5-(tert-butyl)-1,3,4- 0.12 0.016
oxadiazol-2-yl)amino)-[1,1'-biphenyl]-
4-yl)-2-oxabicyclo[2.2.2]octan-4- yl)acetic acid 21
2-(1-(4-(5-((5-cyclobutyl-1,3,4- 0.36 0.944
oxadiazol-2-yl)amino)pyridin-2-
yl)phenyl)-2-oxabicyclo[2.2.2]octan-4- yl)acetic acid 22
2-(4-(4'-((5-(tert-butyl)-1,3,4- 1.7 0.296
oxadiazol-2-yl)amino)-[1,1'-biphenyl]-
4-yl)-7-oxabicyclo[2.2.1]heptan-1- yl)acetic acid 23
3-(4-(4'-((5-(tert-butyl)-1,3,4- 0.084 0.267
oxadiazol-2-yl)amino)-[1,1'-biphenyl]-
4-yl)-7-oxabicyclo[2.2.1]heptan-1- yl)propanoic acid
CONCLUSIONS
[0411] It can be seen that the compounds of the invention are
useful as inhibitors of DGAT1 and therefore useful in the treatment
of diseases and conditions mediated by DGAT1 such as the metabolic
disorders disclosed herein.
[0412] It will be understood that the invention has been described
by way of example only and modifications may be made whilst
remaining within the scope and spirit of the invention.
* * * * *