U.S. patent application number 12/446594 was filed with the patent office on 2010-05-13 for farnesoid x receptor agonists.
This patent application is currently assigned to SMITHKLINE BEECHAM CORPORATION. Invention is credited to Jonathan York Bass, III, Justin Caravella, David Norman Deaton, Robert Blount McFadyen, Frank Navas, III, Paul Kenneth Spearing.
Application Number | 20100120775 12/446594 |
Document ID | / |
Family ID | 39325319 |
Filed Date | 2010-05-13 |
United States Patent
Application |
20100120775 |
Kind Code |
A1 |
Bass, III; Jonathan York ;
et al. |
May 13, 2010 |
Farnesoid X Receptor Agonists
Abstract
The present invention provides novel substituted isoxazole
compounds, pharmaceutical compositions, therapeutic uses and
processes for preparing the same.
Inventors: |
Bass, III; Jonathan York;
(Irvine, CA) ; Deaton; David Norman; (Durham,
NC) ; Caravella; Justin; (Durham, NC) ;
McFadyen; Robert Blount; (Durham, NC) ; Navas, III;
Frank; (Durham, NC) ; Spearing; Paul Kenneth;
(Durham, NC) |
Correspondence
Address: |
GLAXOSMITHKLINE;CORPORATE INTELLECTUAL PROPERTY, MAI B482
FIVE MOORE DR., PO BOX 13398
RESEARCH TRIANGLE PARK
NC
27709-3398
US
|
Assignee: |
SMITHKLINE BEECHAM
CORPORATION
Philadelphia
PA
|
Family ID: |
39325319 |
Appl. No.: |
12/446594 |
Filed: |
October 23, 2007 |
PCT Filed: |
October 23, 2007 |
PCT NO: |
PCT/US07/82170 |
371 Date: |
April 22, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60853886 |
Oct 24, 2006 |
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60855337 |
Oct 30, 2006 |
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60911954 |
Apr 16, 2007 |
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Current U.S.
Class: |
514/249 ;
514/266.2; 514/314; 544/284; 544/355; 546/170 |
Current CPC
Class: |
A61P 3/00 20180101; A61P
3/10 20180101; A61P 1/16 20180101; A61P 3/04 20180101; A61P 1/00
20180101; C07D 413/12 20130101; A61P 3/06 20180101; A61P 17/02
20180101; C07D 413/14 20130101; A61P 11/00 20180101; A61P 13/12
20180101; A61P 43/00 20180101 |
Class at
Publication: |
514/249 ;
546/170; 514/314; 544/284; 514/266.2; 544/355 |
International
Class: |
A61K 31/498 20060101
A61K031/498; C07D 215/20 20060101 C07D215/20; A61K 31/47 20060101
A61K031/47; C07D 413/02 20060101 C07D413/02; A61K 31/517 20060101
A61K031/517; C07D 241/36 20060101 C07D241/36; A61P 3/04 20060101
A61P003/04; A61P 3/10 20060101 A61P003/10; A61P 1/16 20060101
A61P001/16 |
Claims
1. A compound of formula (I): ##STR00282## wherein: each of
Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 is the same or different and
is independently selected from N, CH and C--R.sup.1, wherein one or
two of Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 is N, and at least one
of Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 is C--R.sup.1; each
R.sup.1 is the same or different and is independently selected from
alkyl, fluoroalkyl, --CO.sub.2H, --C(O)NH.sub.2, --CO.sub.2alkyl,
and an acid equivalent group; wherein at least one R.sup.1 is
--CO.sub.2H, --C(O)NH.sub.2, --CO.sub.2alkyl, or an acid equivalent
group; R.sup.2 is H, halo, alkyl or fluoroalkyl; a is 0, 1 or 2;
each R.sup.3 is the same or different and is independently selected
from halo, alkyl and fluoroalkyl; Z.sup.1 is --O--, --S-- or
--N(R.sup.8)--, wherein R.sup.8 is H or alkyl; b is 1, 2 or 3
R.sup.4 is selected from alkyl, 2,2,2-trifluoroethyl,
C.sub.3-6cycloalkyl, alkenyl, C.sub.3-6cycloalkenyl and
fluoro-substituted C.sub.3-6cycloalkyl, c and d are both 0 or c is
1 and d is 0 or 1; R.sup.5 is --C.sub.1-3alkylene-, Z.sup.2 is
--O--, --S(O).sub.e--, or --NH--, wherein e is 0, 1 or 2; Ring D is
C.sub.3-6cycloalkyl or C.sub.3-6cycloalkenyl, optionally
substituted one, two or times with alkyl or fluoroalkyl, or Ring D
is a moiety of formula i, ii or iii: ##STR00283## wherein: f is 1,
2 or 3; and each R.sup.6 is the same or different and is
independently selected from halo, alkyl, fluoroalkyl, --O-alkyl,
--O-fluoroalkyl, alkyl-OH, and alkenyl; or a pharmaceutically
acceptable salt or solvate thereof.
2. A compound of formula (I): ##STR00284## wherein: each of
Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 is the same or different and
is independently selected from N, CH and C--R.sup.1, wherein one of
Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 is N, and at least one of
Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 is C--R.sup.1; each R.sup.1
is the same or different and is independently selected from alkyl,
fluoroalkyl, --CO.sub.2H, --C(O)NH.sub.2, --CO.sub.2alkyl, and an
acid equivalent group; wherein at least one R.sup.1 is --CO.sub.2H,
--C(O)NH.sub.2, --CO.sub.2alkyl, or an acid equivalent group;
R.sup.2 is H, halo, alkyl or fluoroalkyl; a is 0, 1 or 2; each
R.sup.3 is the same or different and is independently selected from
halo, alkyl and fluoroalkyl; Z.sup.1 is --O--, --S-- or
--N(R.sup.8)--, wherein R.sup.8 is H or alkyl; b is 1, 2 or 3
R.sup.4 is selected from alkyl, 2,2,2-trifluoroethyl,
C.sub.3-6cycloalkyl, alkenyl, C.sub.3-6cycloalkenyl and
fluoro-substituted C.sub.3-6cycloalkyl; c and d are both 0 or c is
1 and d is 0 or 1; R.sup.5 is --C.sub.1-3alkylene-; Z.sup.2 is
--O--, --S(O).sub.e--, or --NH--, wherein e is 0, 1 or 2; Ring D is
C.sub.3-6cycloalkyl or C.sub.3-6cycloalkenyl, optionally
substituted one, two or times with alkyl or fluoroalkyl, or Ring D
is a moiety of formula i, ii or iii: ##STR00285## wherein: f is 1,
2 or 3; and each R.sup.6 is the same or different and is
independently selected from halo, alkyl, fluoroalkyl, --O-alkyl,
--O-fluoroalkyl, alkyl-OH, and alkenyl; or a pharmaceutically
acceptable salt or solvate thereof.
3. The compound according to claim 1, wherein Y.sup.4 is N and
Y.sup.3 is C--R.sup.1.
4. The compound according to claim 1, wherein Y.sup.2 is N and
Y.sup.3 C--R.sup.1.
5. The compound according to claim 1, wherein Y.sup.4 is N and
Y.sup.1 and Y.sup.3 are C--R.sup.1.
6. The compound according to claim 1, wherein each R.sup.1 is the
same or different and is independently selected from alkyl,
fluoroalkyl, --CO.sub.2H, --C(O)NH.sub.2 and --CO.sub.2alkyl;
wherein at least one R.sup.1 is --CO.sub.2H, --C(O)NH.sub.2 or
--CO.sub.2alkyl.
7. The compound according to claim 1, wherein at least one R.sup.1
is --CO.sub.2H or --CO.sub.2alkyl.
8. The compound according to claim 1, wherein R.sup.2 is H.
9. The compound according to claim 1, wherein a is 0.
10. The compound according to claim 1, wherein Z.sup.1 is
--O--.
11. The compound according to claim 1, wherein b is 1.
12. The compound according to claim 1, wherein R.sup.4 is alkyl or
2,2,2-trifluoroethyl or C.sub.2-6cycloalkyl.
13. The compound according to claim 1, wherein R.sup.4 is
isopropyl.
14. The compound according to claim 1, wherein c and d are both
0.
15. The compound according to claim 1, wherein c is 1 and R.sup.5
is methylene or ethylene.
16. The compound according to claim 1, wherein c is 1, d is 1 and
Z.sup.2 is --O--.
17. The compound according to claim 1, wherein Ring D is a moiety
of formula i, ii or iii: ##STR00286##
18. The compound according to claim 1, wherein Ring D is
##STR00287## and f is 2.
19. The compound according to claim 1 wherein Ring D is a moiety of
formula i, ii or iii and each R.sup.6 is the same and is halo or
alkyl.
20. A compound selected from
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2- quinolinecarboxylic acid;
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl}-3-quinolinecarboxylic acid;
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2,4-quinolinedicarboxylic acid;
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-quinolinecarboxylic acid;
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid;
6-[4-({[5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylic acid;
6-[4-({[5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylic acid
6-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-2-quinolinecarboxylic acid;
7-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-3-isoquinolinecarboxylic acid;
7-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylic acid;
6-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylic acid;
6-[4-({[5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}ox-
y)phenyl]-2-quinolinecarboxylic acid;
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylic acid;
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylic acid;
6-[4-({3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propyl}ox-
y)phenyl]-2-quinolinecarboxylic acid;
7-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-3-isoquinolinecarboxylic acid;
7-(4-{[(5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)met-
hyl]oxy}phenyl)-3-isoquinolinecarboxylic acid;
6-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylic acid;
6-(4-{[(5-(1-methylethyl)-3-{[(2,4,6-trifluorophenyl)oxy]methyl}-4-isoxaz-
olyl)methyl]oxy}phenyl)-2-quinolinecarboxylic acid;
6-[4-({[3-{[(2,6-dichlorophenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-{[(2,6-dichlorophenyl)amino]methyl}-5-(1-methylethyl)-4-isoxazo-
lyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazol-
yl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-{[(2,6-dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-{[(2,6-dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid;
6-(4-{[(3,5-dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinolinecarb-
oxylic acid;
6-[4-({[34(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methyl}o-
xy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-(2,4-dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-7-fluoro-2-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-methyl-2-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-3-methylphenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-2-methylphenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-methyl-2-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-methyl-3-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-1-isoquinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4- methyl-2-quinazolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio-
)phenyl]-2-quinolinecarboxylic acid;
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinoxalinecarboxylic acid; and pharmaceutically
acceptable salts or solvates thereof.
21.
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylic acid or a pharmaceutically
acceptable salt or solvate thereof.
22.
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylic acid.
23.
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylic acid potassium salt.
24. A pharmaceutical composition comprising a compound according to
claim 1 and a pharmaceutically acceptable carrier or diluent.
25. A pharmaceutical composition comprising
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid or a pharmaceutically acceptable
salt or solvate thereof and a pharmaceutically acceptable carrier
or diluent.
26. A method for the treatment of a condition mediated by decreased
FXR activity in a subject in need thereof, said method comprising
administering to said subject a therapeutically effective amount of
a compound according to claim 1.
27. A method for the treatment of obesity in a mammal in need
thereof comprising administering to said subject a therapeutically
effective amount of a compound according to claim 1.
28. A method for the treatment of diabetes mellitus in a mammal in
need thereof comprising administering to said subject a
therapeutically effective amount of a compound according to claim
1.
29. A method for the treatment of metabolic syndrome in a mammal in
need thereof comprising administering to said subject a
therapeutically effective amount of a compound according to claim
1.
30. A method for the treatment of cholestatic liver disease in a
mammal in need thereof comprising administering to said subject a
therapeutically effective amount of a compound according to claim
1.
31. A method for the treatment of organ fibrosis in a mammal in
need thereof comprising administering to said subject a
therapeutically effective amount of a compound according to claim
1.
32. A method for the treatment of liver fibrosis in a mammal in
need thereof, said method comprising administering to said subject
a therapeutically effective amount of a compound according to claim
1.
33. A method for the treatment of diabetes mellitus in a mammal in
need thereof comprising administering to said subject a
therapeutically effective amount of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid or a pharmaceutically acceptable
salt or solvate thereof.
34. A method for the treatment of cholestatic liver disease in a
mammal in need thereof comprising administering to said subject a
therapeutically effective amount of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid or a pharmaceutically acceptable
salt or solvate thereof.
35. A process for preparing a compound according to claim 1
comprising the steps of: a) reacting a compound of formula (II)
##STR00288## with a compound of formula (III) ##STR00289## wherein:
X.sup.1 is chloride, iodide, bromide, triflate, tosylate, nosylate,
besylate or mesylate, (preferably chloro); each R.sup.1 is the same
or different and is independently selected from alkyl, fluoroalkyl
or --CO.sub.2alkyl; wherein at least one R.sup.1 is
--CO.sub.2alkyl; and all other variables are as defined above for
formula (I) to prepare a compound of formula (I).
36-51. (canceled)
Description
BACKGROUND OF THE INVENTION
[0001] The present invention relates to farnesoid X receptors (FXR,
NR1H4). More particularly, the present invention relates to
compounds useful as agonists for FXR, pharmaceutical formulations
comprising such compounds, and therapeutic use of the same.
[0002] FXR is a member of the nuclear receptor class of
ligand-activated transcription factors. Physiological
concentrations of bile acids bind and activate FXR. [Parks, D. J.,
et al. 1999 Science 284:1365-1368; Makishima, M., et al. 1999
Science 284:1362-1365] Bile acids are amphipathic molecules that
form micelles and emulsify dietary lipids. This property also makes
bile acids cytotoxic if sufficient concentrations are achieved and
thus mechanisms have evolved to ensure bile acid concentrations are
tightly regulated. FXR plays a key role in regulating bile acid
homeostasis. [Makishima, M. 2005 J. Pharmacol. Sci. 97:177-183;
Kuipers, F., et al. 2004 Rev. Endocrine Metab. Disorders
5:319-326]
[0003] FXR is expressed in liver, intestine, kidney, and adrenal.
[Kuipers, F., et al. 2004 Rev. Endocrine Metab. Disorders
5:319-326] FXR target genes in hepatocytes include small
heterodimer partner (SHP, NR0B2) which encodes an atypical nuclear
receptor that represses transcription of genes such as CYP7A1
(encoding cholesterol 7.alpha.-hydroxylase), the first and rate
limiting step in the conversion of cholesterol to bile acid, CYP8B1
(encoding sterol 12.alpha.-hydroxylase) which controls the
hydrophobicity of the bile pool and NTCP (encoding the
sodium/taurocholate co-transporting polypeptide, SLC10A1) that
imports bile acids from the portal and systemic circulation into
the hepatocyte. [Goodwin, B., et al. 2000 Mol. Cell 6:517-526; del
Castillo-Olivares, A., et al 2001 Nucleic Acids Res. 29:4035-4042;
Denson, L. A., et al. 2001 Gastroenterology 121(1):140-147] Other
FXR target genes that are induced in liver include the canalicular
transporter BSEP (encoding the bile salt export pump, ABCB11) that
transports bile acids from the hepatocyte into the bile, multi-drug
resistance P glycoprotein-3 (MDR3) (encoding the canalicular
phospholipid flippase, ABCB4) that transports phospholipids from
the hepatocyte into the bile and MRP2 (encoding multidrug
resistance-related protein-2, ABCC2) that transports conjugated
bilirubin, glutathione and glutathione conjugates into bile.
[Ananthanarayanan, M., et al. 2001 J. Biol. Chem. 276:28857-28865;
Huang, L et al., 2003 J. Biol. Chem. 278:51085-51090; Kast, H. R.,
et al. 2002 J. Biol. Chem. 277:2908-2915.]
[0004] In the intestine FXR also induces expression of SHP which
represses transcription of the apical sodium dependent bile acid
transporter (ASBT, SLC10A2) gene which encodes the high affinity
apical sodium dependent bile acid transporter that moves bile acids
from the intestinal lumen into the enterocyte as part of the
enterohepatic recycling of bile acids. [Li, H., et al. 2005 Am. J.
Physiol. Gastrointest. Liver Physiol. 288:G60-G66] Ileal bile acid
binding protein (IBABP) gene expression is also induced by FXR
agonists in the enterocyte. [Grober, J. et al., 1999 J. Biol. Chem.
274:29749-29754] The function of this ileal bile acid binding
protein remains under investigation.
[0005] Cholestasis is a condition of reduced or arrested bile flow.
Unresolved cholestasis leads to liver damage such as that seen in
primary biliary cirrhosis (PBC) and primary sclerosing cholangitis
(PSC), two cholestatic liver diseases. FXR agonists have been shown
to protect the liver in rodent models of cholestatic liver disease.
[Liu, Y. et al. 2003 J. Clin. Invest. 112:1678-1687; Fiorucci, S.,
et al. 2005 J. Pharmacol. Exp. Ther. 313:604-612; Pellicciari, R.,
et al. 2002 J. Med. Chem. 45:3569-3572]
[0006] FXR is also expressed in hepatic stellate cells (HSC) which
play a role in deposition of extracellular matrix during the
fibrotic process. Treatment of cultured HSCs with the FXR agonist
6-ethyl-chenodeoxycholic acid (6EtCDCA) results in decreased
expression of fibrotic markers such as .alpha.-smooth muscle actin
and .alpha.1(I)collagen. 6EtCDCA has also been reported to prevent
development and promote resolution of hepatic fibrosis in multiple
rodent models of this disease. [Fiorucci, S., et al., 2004
Gastroenterology 127:1497-1512; Fiorucci, S., et al., 2005 J.
Pharmacol. Exp. Ther. 314:584-595.] According to Fiorucci et al.,
this anti-fibrotic effect is due to SHP inactivation of Jun and
subsequent repression of tissue inhibitor of metalloproteinase 1
(TIMP1) via the activation protein 1 (AP1) binding site on the
TIMP1 promoter.
[0007] Recently, S. Kliewer presented data at the Digestive
Diseases Week (DDW) Conference (2005) organized by the American
Association for the study of Liver Disease (AASLD) showing that
activation of FXR by the agonist GW4064 resulted in improved
mucosal barrier and decreased bacterial overgrowth in a bile
duct-ligated mouse model of cholestasis and intestinal bacterial
overgrowth. Dr. Kliewer showed data indicating decreased
translocation of bacteria to mesenteric lymph nodes in mice treated
with GW4064. This effect of GW4064 was lost in FXR null mice.
[Inagaki, T., et al. 2006 Proc. Nat. Acad. Sci., U. S. A.
103:3920-3925.]
[0008] The FXR agonist GW4064, when administered to mice on a
lithogenic diet, prevented the formation of cholesterol crystals in
the bile. This effect of the compound was lost in FXR null mice.
Moschetta, A., et al. 2004 Nat. Med. 10:1352-1358.
[0009] It has been suggested that GW4064 could improve lipid and
glucose homeostasis and insulin sensitivity in rodent diabetic and
insulin resistance models. Chen and colleagues [2006 Diabetes 55
suppl. 1: A200] demonstrated that when administered to mice on
high-fat diet, GW4064 decrease body weight and body fat mass, serum
glucose, insulin, triglyceride, and total cholesterol. GW4064 also
corrected glucose intolerance in those animals. In addition, GW4064
decreased serum insulin concentration, improved glucose tolerance
and enhanced insulin sensitivity in ob/ob mice [Cariou, B., et al.,
2006 J. Biol. Chem. 281:11039-11049]. In another study, it was
reported that GW4064 significantly improved hyperglycemia and
hyperlipidemia in diabetic db/db mice [Zhang, Y., et al. 2006 Proc.
Nat. Acad. Sci., U.S.A. 103:1006-1011].
SUMMARY OF THE INVENTION
[0010] As a first aspect, the present invention provides compounds
of formula (I):
##STR00001## [0011] wherein: [0012] each of Y.sup.1, Y.sup.2,
Y.sup.3 and Y.sup.4 is the same or different and is independently
selected from N, CH and C--R.sup.1, wherein one or two of Y.sup.1,
Y.sup.2, Y.sup.3 and Y.sup.4 is N, and at least one of Y.sup.1,
Y.sup.2, Y.sup.3 and Y.sup.4 is C--R.sup.1; [0013] each R.sup.1 is
the same or different and is independently selected from alkyl,
fluoroalkyl, --CO.sub.2H, --C(O)NH.sub.2, --CO.sub.2alkyl, and an
acid equivalent group; wherein at least one R.sup.1 is --CO.sub.2H,
--C(O)NH.sub.2, --CO.sub.2alkyl, or an acid equivalent group;
[0014] R.sup.2 is H, halo, alkyl or fluoroalkyl; [0015] a is 0, 1
or 2; [0016] each R.sup.3 is the same or different and is
independently selected from halo, alkyl and fluoroalkyl; [0017]
Z.sup.1 is --O--, --S-- or --N(R.sup.8)--, wherein R.sup.8 is H or
alkyl; [0018] b is 1, 2 or 3; [0019] R.sup.4 is selected from
alkyl, 2,2,2-trifluoroethyl, C.sub.3-6cycloalkyl, alkenyl,
C.sub.3-6cycloalkenyl and fluoro-substituted C.sub.3-6cycloalkyl;
[0020] c and d are both 0 or c is 1 and d is 0 or 1; [0021] R.sup.5
is --C.sub.1-3alkylene-; [0022] Z.sup.2 is --O--, --S(O).sub.e--,
or --NH--, wherein e is 0, 1 or 2; [0023] Ring D is
C.sub.3-6cycloalkyl or C.sub.3-6cycloalkenyl, optionally
substituted one, two, or three times with alkyl or fluoroalkyl, or
Ring D is a moiety of formula i, ii or iii:
##STR00002##
[0024] wherein:
[0025] f is 1, 2 or 3; and
[0026] each R.sup.6 is the same or different and is independently
selected from halo, alkyl, fluoroalkyl, --O-alkyl, --O-fluoroalkyl,
alkyl-OH, and alkenyl; [0027] or a pharmaceutically acceptable salt
or solvate thereof.
[0028] In a second aspect, the present invention provides a
pharmaceutical composition comprising a compound of formula (I).
The composition may further comprise a pharmaceutically acceptable
carrier or diluent.
[0029] In a third aspect, the present invention provides a method
for the treatment of a condition mediated by decreased FXR activity
in a subject in need thereof. The method comprises administering to
the subject a therapeutically effective amount of a compound of
formula (I).
[0030] In a fourth aspect, the present invention provides a method
for the treatment of obesity in a subject in need thereof. The
method comprises administering to the subject a therapeutically
effective amount of a compound of formula (I).
[0031] In a fifth aspect, the present invention provides a method
for the treatment of diabetes mellitus in a subject in need
thereof. The method comprises administering to the subject a
therapeutically effective amount of a compound of formula (I).
[0032] In a sixth aspect, the present invention provides a method
for the treatment of metabolic syndrome in a subject in need
thereof. The method comprises administering to the subject a
therapeutically effective amount of a compound of formula (I).
[0033] In a seventh aspect, the present invention provides a method
for the treatment of cholestatic liver disease in a subject in need
thereof. The method comprises administering to the subject a
therapeutically effective amount of a compound of formula (I).
[0034] In a eighth aspect, the present invention provides a method
for the treatment of organ fibrosis in a subject in need thereof.
The method comprises administering to the subject a therapeutically
effective amount of a compound of formula (I). In one embodiment,
the organ fibrosis is liver fibrosis.
[0035] In a ninth aspect, the present invention provides a method
for the treatment of liver fibrosis in a subject in need thereof.
The method comprises administering to the subject a therapeutically
effective amount of a compound of formula (I).
[0036] In a tenth aspect, the present invention provides a process
for preparing a compound of formula (I). The process comprises the
steps of: [0037] a) reacting a compound of formula (II)
[0037] ##STR00003## [0038] with a compound of formula (III)
##STR00004##
[0039] wherein: [0040] X.sup.1 is chloride, iodide, bromide,
triflate, tosylate, nosylate, besylate or mesylate, (preferably
chloro); [0041] each R.sup.1 is the same or different and is
independently selected from alkyl, fluoroalkyl or [0042]
--CO.sub.2alkyl; wherein at least one R.sup.1 is --CO.sub.2alkyl;
and [0043] all other variables are as defined above for formula (I)
to prepare a compound of formula (I); [0044] b) optionally
converting the compound of formula (I) into a pharmaceutically
acceptable salt or solvate thereof; [0045] c) optionally converting
the compound of formula (I) or a pharmaceutically acceptable salt
or solvate thereof into a different compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof.
[0046] In another aspect, the present invention provides another
process for preparing a compound of formula (I). This process
comprises the steps of: [0047] a) reacting a compound of formula
(IV)
[0047] ##STR00005## [0048] with a compound of formula (II) under
Mitsunobu reaction conditions
##STR00006##
[0049] wherein at least one R.sup.1 is --CO.sub.2alkyl and all
other variables are as defined above to prepare a compound of
formula (I); [0050] b) optionally converting the compound of
formula (I) into a pharmaceutically acceptable salt or solvate
thereof; and [0051] c) optionally converting the compound of
formula (I) or a pharmaceutically acceptable salt or solvate
thereof into a different compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof.
[0052] In another aspect, the present invention provides another
process for preparing a compound of formula (I). This process
comprises the steps of: [0053] a) reacting a compound of formula
(XLI) under Mitsunobu reaction conditions
[0053] ##STR00007## [0054] with a compound of formula i-a, ii-a, or
iii-a
##STR00008##
[0055] wherein at least one R.sup.1 is --CO.sub.2alkyl
[0056] Z.sup.2a is selected from --O--, --S--, --N(H)--, and
N(COCF.sub.3); and
[0057] all other variables are as defined above [0058] to prepare a
compound of formula (I-A)
##STR00009##
[0059] wherein Ring D is a moiety of formula i, ii or iii:
##STR00010## [0060] b) optionally converting the compound of
formula (I-A) into a pharmaceutically acceptable salt or solvate
thereof; and [0061] c) optionally converting the compound of
formula (I-A) or a pharmaceutically acceptable salt or solvate
thereof into a different compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof.
[0062] In another aspect, the present invention provides another
process for preparing a compound of formula (I). This process
comprises the steps of: [0063] a) reacting a compound of formula
(XXV)
##STR00011##
[0064] wherein: X.sup.2 is chloro, bromo, iodo, or triflate;
[0065] and at least one R.sup.1 is --CO.sub.2alkyl; [0066] with a
boronic acid or ester compound of formula (XLV) under Suzuki
coupling conditions
##STR00012##
[0067] wherein: [0068] R.sup.10 is H or alkyl; and [0069] all other
variables are as defined above [0070] to prepare a compound of
formula (I); [0071] b) optionally converting the compound of
formula (I) into a pharmaceutically acceptable salt or solvate
thereof; and [0072] c) optionally converting the compound of
formula (I) or a pharmaceutically acceptable salt or solvate
thereof into a different compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof.
[0073] In another aspect, the present invention provides another
process for preparing a compound of formula (I). The process
comprises the steps of: [0074] a) reacting a compound of formula
(XXV-5)
[0074] ##STR00013## [0075] with a boronic acid or ester of compound
of formula (XLV) under Suzuki coupling conditions
##STR00014##
[0076] wherein at least one R.sup.1 is C.sub.1-6alkyl or
fluoroalkyl;
[0077] R.sup.10 is H or alkyl; and
[0078] all other variables are as defined above to prepare a
compound of formula (I) wherein Y.sup.2 is N and Y.sup.4 is N;
[0079] b) optionally converting the compound of formula (I) into a
pharmaceutically acceptable salt or solvate thereof; and [0080] c)
optionally converting the compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof into a
different compound of formula (I) or a pharmaceutically acceptable
salt or solvate thereof.
[0081] In another aspect, the present invention provides another
process for preparing a compound of formula (I). The process
comprises the steps of: [0082] a) reacting a compound of formula
(XXV-6)
##STR00015##
[0083] with a boronic acid or ester of compound of formula (XLV)
under Suzuki coupling conditions wherein at least one R.sup.1 is
C.sub.1-6alkyl or fluoroalkyl;
[0084] R.sup.10 is H or alkyl; and
[0085] all other variables are as defined above [0086] to prepare a
compound of formula (I) wherein Y.sup.1 is N and Y.sup.4 is N;
[0087] b) optionally converting the compound of formula (I) into a
pharmaceutically acceptable salt or solvate thereof; and [0088] c)
optionally converting the compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof into a
different compound of formula (I) or a pharmaceutically acceptable
salt or solvate thereof.
[0089] In another aspect, the present invention provides a compound
of formula (I) for use in therapy. The present invention also
provides a compound of formula (I) for use in the treatment of a
condition mediated by decreased FXR activity in a subject; a
compound of formula (I) for use in the treatment of obesity in a
subject; a compound of formula (I) for use in the treatment of
diabetes mellitus in a subject; a compound of formula (I) for use
in the treatment of metabolic syndrome in a subject; a compound of
formula (I) for use in the treatment of cholestatic liver disease
in a subject; a compound of formula (I) for use in the treatment of
organ fibrosis in a subject; and a compound of formula (I) for use
in the treatment of liver fibrosis in a subject.
[0090] In another aspect, the present invention provides the use of
a compound of formula (I) for the preparation of a medicament for
the treatment of a condition mediated by decreased FXR activity in
a subject; the use of a compound of formula (I) for the preparation
of a medicament for the treatment of obesity; the use of a compound
of formula (I) for the preparation of a medicament for the
treatment of diabetes mellitus in a subject; the use of a compound
of formula (I) for the preparation of a medicament for the
treatment of metabolic syndrome in a subject; the use of a compound
of formula (I) for the preparation of a medicament for the
treatment of cholestatic liver disease in a subject; the use of a
compound of formula (I) for the preparation of a medicament for the
treatment of organ fibrosis in a subject; and the use of a compound
of formula (I) for the preparation of a medicament for the
treatment of liver fibrosis in a subject.
[0091] In another aspect, the present invention provides a
pharmaceutical composition comprising a compound of formula (I) for
use in the treatment of a condition mediated by decreased FXR
activity.
[0092] Further aspects of the present invention are described in
the description of particular embodiments, examples, and claims
which follow.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0093] As used herein, "a compound of the invention" or "a compound
of formula (I)" or "(I-A)," etc. means a compound of formula (I)
(or (I-A)) or a pharmaceutically acceptable salt or solvate thereof
Similarly, with respect to isolatable intermediates such as for
example, compounds of formula (II), (III), (IV), (V), (XL), (XLI)
and (XLII), the phrase "a compound of formula (number)" means a
compound having that formula or a pharmaceutically acceptable salt
or solvate thereof
[0094] As used herein, the term "alkyl" refers to aliphatic
straight or branched saturated hydrocarbon chains containing 1-8
carbon atoms. Examples of "alkyl" groups as used herein include but
are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, t-butyl, pentyl, hexyl, octyl and the like.
[0095] The term "fluoroalkyl" as used herein refers to an alkyl as
defined above substituted with one or more fluoro. In on particular
embodiment, fluoroalkyl refers to an alkyl substituted with two or
more fluoro. The term "di(fluoroalkyl)" refers to two alkyl as
defined above each substituted with one or more fluoro. For
example, the term "di(fluoromethyl)" refers to
--(CH.sub.2F).sub.2.
[0096] The term "alkylene" refers to a straight or branched alkyl
bridge, i.e., the group -alkyl-, wherein alkyl is as defined
above.
[0097] As used herein, the term "halo" refers to any halogen atom.
i.e., fluorine, chlorine, bromine or iodine.
[0098] As used herein, the term "alkenyl" refers to an aliphatic
straight or branched unsaturated hydrocarbon chain containing 2-8
carbon atoms and at least one and up to three carbon-carbon double
bonds. Examples of "alkenyl" groups as used herein include but are
not limited to ethenyl and propenyl.
[0099] The term "alkenylene" refers to a straight or branched
alkenyl bridge, i.e., the group -alkenyl-, wherein alkenyl is as
defined above.
[0100] As used herein, the term "cycloalkyl" refers to a
non-aromatic monocyclic carbocyclic ring having from 3 to 8 carbon
atoms (unless a different number of atoms is specified) and no
carbon-carbon double bonds. "Cycloalkyl" includes by way of example
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and
cyclooctyl. Particular cycloalkyl groups include
C.sub.3-6cycloalkyl.
[0101] As used herein, the term "cycloalkenyl" refers to a
non-aromatic monocyclic carbocyclic ring having from 3 to 8 carbon
atoms (unless a different number of atoms is specified) and from 1
to 3 carbon-carbon double bonds. "Cycloalkenyl" includes by way of
example cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl,
cycloheptenyl and cyclooctenyl. Particular cycloalkenyl groups
include C.sub.3-6cycloalkenyl.
[0102] As used herein, the term "optionally" means that the
subsequently described event(s) may or may not occur, and includes
both event(s) that occur and events that do not occur.
[0103] The present invention relates to compounds of formula
(I):
##STR00016## [0104] wherein: [0105] each of Y.sup.1, Y.sup.2,
Y.sup.3 and Y.sup.4 is the same or different and is independently
selected from N, CH and C--R.sup.1, wherein one or two of Y.sup.1,
Y.sup.2, Y.sup.3 and Y.sup.4 is N, and at least one of Y.sup.1,
Y.sup.2, Y.sup.3 and Y.sup.4 is C--R.sup.1; [0106] each R.sup.1 is
the same or different and is independently selected from alkyl,
fluoroalkyl, --CO.sub.2H, --C(O)NH.sub.2, --CO.sub.2alkyl, and an
acid equivalent group; wherein at least one R.sup.1 is --CO.sub.2H,
--C(O)NH.sub.2, --CO.sub.2alkyl, or an acid equivalent group;
[0107] R.sup.2 is H, halo, alkyl or fluoroalkyl; [0108] a is 0, 1
or 2; [0109] each R.sup.3 is the same or different and is
independently selected from halo, alkyl and fluoroalkyl; [0110]
Z.sup.1 is --O--, --S-- or --N(R.sup.8)--, wherein R.sup.8 is H or
alkyl; [0111] b is 1, 2 or 3 [0112] R.sup.4 is selected from alkyl,
2,2,2-trifluoroethyl, C.sub.3-6cycloalkyl, alkenyl,
C.sub.3-6cycloalkenyl and fluoro-substituted C.sub.3-6cycloalkyl;
[0113] c and d are both 0 or c is 1 and d is 0 or 1; [0114] R.sup.5
is --C.sub.1-3alkylene-; [0115] Z.sup.2 is --O--, --S(O).sub.e--,
or --NH--, wherein e is 0, 1 or 2; [0116] Ring D is
C.sub.3-6cycloalkyl or C.sub.3-6cycloalkenyl, optionally
substituted one, two, or three times with alkyl or fluoroalkyl, or
Ring D is a moiety of formula i, ii or iii:
##STR00017##
[0117] wherein:
[0118] f is 1, 2 or 3; and
[0119] each R.sup.6 is the same or different and is independently
selected from halo, alkyl, fluoroalkyl, --O-alkyl, --O-fluoroalkyl,
alkyl-OH, and alkenyl; [0120] or a pharmaceutically acceptable salt
or solvate thereof.
[0121] Any one or two of Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 may
be N. In one particular embodiment only one of Y.sup.1, Y.sup.2,
Y.sup.3 and Y.sup.4 is N. At least one of Y.sup.1, Y.sup.2, Y.sup.3
and Y.sup.4 is C--R.sup.1. In a particular embodiment, two of
Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 are independently C--R.sup.1.
In another particular embodiment, only one of Y.sup.1, Y.sup.2,
Y.sup.3 and Y.sup.4 is C--R.sup.1. In such embodiment one or two of
the other variables Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4 is N and
the remaining other variables Y.sup.1, Y.sup.2, Y.sup.3 and Y.sup.4
are CH.
[0122] In one embodiment, the compounds of formula (I) are defined
wherein Y.sup.2 is N and Y.sup.3 is C--R.sup.1. In one particular
version of this embodiment, Y.sup.1 and Y.sup.4 are selected from
CH and C--R.sup.1, more particularly both are CH. In another
embodiment, the compounds of formula (I) are defined wherein
Y.sup.1 is N and Y.sup.3 is C--R.sup.1. In one particular version
of this embodiment, Y.sup.2 and Y.sup.4 are both CH. In another
embodiment, the compounds of formula (I) are defined wherein
Y.sup.1 is N and Y.sup.4 is C--R.sup.1. In one particular version
of this embodiment, Y.sup.2 and Y.sup.3 are CH. In another
embodiment, the compounds of formula (I) are defined wherein
Y.sup.3 is N and Y.sup.4 is C--R.sup.1. In another embodiment, the
compounds of formula (I) are defined wherein Y.sup.4 is N and
Y.sup.2 is C--R.sup.1. In one particular version of this
embodiment, Y.sup.1 is CH and Y.sup.3 is C--R.sup.1. In one
preferred embodiment, the compounds of formula (I) are defined
wherein Y.sup.4 is N and Y.sup.3 is C--R.sup.1. In such embodiment,
Y.sup.1 and Y.sup.2 are preferably selected from CH and C--R.sup.1,
more particularly CH.
[0123] In an additional embodiment, the compounds of formula (I)
are defined wherein Y.sup.2 and Y.sup.4 are N. In one particular
version of this embodiment, Y.sup.1 and Y.sup.3 are C--R.sup.1.
[0124] Specific examples of the bicyclic fused N-heteroaryl
moiety:
##STR00018## [0125] include but are not limited to the
following:
[0125] ##STR00019## [0126] wherein each R.sup.1 is the same or
different and is independently selected from --CO.sub.2H,
--C(O)NH.sub.2, --CO.sub.2alkyl or acid equivalent group.
[0127] In one preferred embodiment, the bicyclic fused N-heteroaryl
moiety is:
##STR00020##
[0128] In another preferred embodiment, the bicyclic fused
N-heteroaryl moiety is:
##STR00021##
[0129] In third preferred embodiment, the bicyclic fused
N-heteroaryl moiety is:
##STR00022##
[0130] In a fourth preferred embodiment, the bicyclic fused
N-heteroaryl moiety is:
##STR00023##
[0131] In a fifth preferred embodiment, the the bicyclic fused
N-heteroaryl moiety is:
##STR00024##
[0132] In all embodiments, at least one of R.sup.1 is --CO.sub.2H,
--C(O)NH.sub.2, --CO.sub.2alkyl or acid equivalent group. In one
embodiment, the compound of formula (I) is defined wherein each
R.sup.1 is the same or different and is independently selected from
alkyl, fluoroalkyl, --CO.sub.2H, --C(O)NH.sub.2 and
--CO.sub.2alkyl. In such embodiment, at least one R.sup.1 is
--CO.sub.2H, --C(O)NH.sub.2 or --CO.sub.2alkyl. In one preferred
embodiment, at least one R.sup.1 is --CO.sub.2H or --CO.sub.2alkyl,
such as --CO.sub.2CH.sub.3.
[0133] In one embodiment of the invention, R.sup.2 is H.
[0134] In one embodiment, a is 0. In the embodiment, wherein a is
1, R.sup.3 is preferably halo (particularly F or Cl), CH.sub.3,
CF.sub.3, or CH.sub.2CH.sub.3.
[0135] In one embodiment, Z.sup.1 is --O--, --S-- or --N(H)--. In
one preferred embodiment, Z.sup.1 is O.
[0136] In one embodiment, b is 1 or 3. In one preferred embodiment,
b is 1.
[0137] In one embodiment, R.sup.4 is linear or branched alkyl or
2,2,2-trifluoroethyl or C.sub.3-6cycloalkyl. Specific examples of
groups defining R.sup.4 include but are not limited to methyl,
ethyl, 2,2,2-trifluoroethyl, propyl, isopropyl, n-butyl, t-butyl,
isobutyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. In
one embodiment, R.sup.4 is isopropyl, isobutyl, cyclopropyl,
cyclobutyl, cyclopentyl or trifluoroethyl. In one embodiment,
R.sup.4 is isopropyl, isobutyl, cyclopropyl or cyclobutyl. In one
particular embodiment, R.sup.4 is isopropyl or isobutyl. In one
preferred embodiment, R.sup.4 is isopropyl.
[0138] The invention includes compounds of formula I' wherein c and
d are both 0 and thus Ring D is bound directly to the isoxazole
ring as shown in formula (I'):
##STR00025## [0139] wherein all other variables are as defined
above.
[0140] The invention includes of formula (I'') wherein c is 1 and d
is 0 or 1 and thus Ring D is bound to C.sub.1-3 alkylene (R.sup.5)
or Z.sup.2 (when d is 1) as shown in formula (I'').
##STR00026## [0141] wherein all other variables are as defined
above.
[0142] In one embodiment, wherein c is 1, R.sup.5 is preferably
methylene or ethylene. In the embodiment wherein both c and d are
1, R.sup.5 is preferably methylene. In one embodiment, c is 1, d is
1 and Z.sup.2 is O. In one particular embodiment, c is 1, d is 1,
R.sup.5 is methylene and Z.sup.2 is O, as in formula (I''').
##STR00027## [0143] wherein all other variables are as defined
above. The invention includes compounds of formula I'''.
[0144] Ring D is C.sub.3-6cycloalkyl or C.sub.3-6cycloalkenyl,
optionally substituted one, two, or three times with alkyl or
fluoroalkyl, or Ring D is a moiety of formula i, ii or iii:
##STR00028##
[0145] wherein:
[0146] f is 1, 2 or 3; and
[0147] each R.sup.6 is the same or different and is independently
selected from halo, alkyl, fluoroalkyl, --O-alkyl, --O-fluoroalkyl,
alkyl-OH, and alkenyl;
[0148] In the embodiment wherein D is C.sub.3-6cycloalkyl or
C.sub.3-6cycloalkenyl optionally substituted one, two, or three
times with alkyl or fluoroalkyl, typically at least one alkyl or
fluoroalkyl substituent is adjacent to the point of attachment of
the cycloalkyl or cycloakenyl ring to the isoxazole R, R.sup.5 or
Z.sup.2 as the case may be. The following specific (but not
limiting) examples a, b, and c illustrate at least one substituent
adjacent to the point of attachment:
##STR00029##
[0149] In one embodiment, Ring D is C.sub.3-6cycloalkyl or
C.sub.3-6cycloalkenyl, optionally substituted symmetrically with
dialkyl or di(fluoroalkyl). Example a above illustrates a
C.sub.3-.sub.6cycloalkyl substituted symmetrically. More
particularly in one embodiment, Ring D is C.sub.3-6cycloalkyl or
C.sub.3-6cycloalkenyl, optionally substituted symmetrically with
di(fluoromethyl).
[0150] In one embodiment, Ring D is a moiety of formula i, ii or
iii. In one particular embodiment, Ring D is a moiety of formula i
or ii. In the embodiment wherein Ring D is
##STR00030##
f is preferably 2. More particularly, in one embodiment f is 2 and
Ring D is a moiety of formula i-b:
##STR00031##
[0151] In one embodiment, the compound of formula (I) is defined
wherein Ring D is a moiety of formula i, ii or iii and each R.sup.6
is the same and is halo or alkyl. In one particular embodiment,
Ring D is a moiety of formula i, ii or iii and each R.sup.6 is the
same and is F, Cl or methyl. In one preferred embodiment, Ring D
is
##STR00032##
f is 2 and each R.sup.6 is Cl.
[0152] In one particular preferred embodiment, Ring D is a moiety
of formula i-b
##STR00033##
and each R.sup.6 is Cl.
[0153] In one preferred embodiment, the invention provides
compounds of formula (I-B):
##STR00034## [0154] wherein [0155] R.sup.1 is --CO.sub.2H,
--C(O)NH.sub.2, --CO.sub.2alkyl, or an acid equivalent group; more
particularly --CO.sub.2H, --C(O)NH.sub.2 or --CO.sub.2alkyl; [0156]
R.sup.4 is isopropyl, isobutyl, cyclopropyl or cyclobutyl; and
[0157] both R.sup.6 are the same and are selected from F, Cl,
CH.sub.3 and CF.sub.3.
[0158] In another preferred embodiment, the invention provides
compounds of formula (I-Z)
##STR00035## [0159] wherein [0160] R.sup.1 is --CO.sub.2H,
--C(O)NH.sub.2, --CO.sub.2alkyl, or an acid equivalent group; more
particularly --CO.sub.2H, --C(O)NH.sub.2 or --CO.sub.2alkyl; [0161]
R.sup.4 is isopropyl, isobutyl, cyclopropyl or cyclobutyl; and
[0162] both R.sup.6 are the same and are selected from F, Cl,
CH.sub.3 and CF.sub.3.
[0163] The present invention contemplates and includes all
combinations and subsets of the particular groups defined
above.
[0164] Specific examples of particular compounds of the present
invention are selected from the group consisting of: [0165]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid; [0166]
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-quinolinecarboxylic acid; [0167]
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2,4-quinolinedicarboxylic acid; [0168]
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-quinolinecarboxylic acid; [0169]
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid; [0170]
6-[4-({[5-Cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylic acid; [0171]
6-[4-({[5-Cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylic acid [0172]
6-[4-({[5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-2-quinolinecarboxylic acid; [0173]
7-[4-({[5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-3-isoquinolinecarboxylic acid; [0174]
7-[4-({[5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylic acid; [0175]
6-[4-({[5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylic acid; [0176]
6-[4-({[5-Cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}ox-
y)phenyl]-2-quinolinecarboxylic acid; [0177]
6-{4-[({3-(2,6-Dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylic acid; [0178]
6-{4-[({3-(2,6-Dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylic acid; [0179]
6-[4-({3-[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propyl}ox-
y)phenyl]-2-quinolinecarboxylic acid; [0180]
7-[4-({[3-{[(2,6-Dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-3-isoquinolinecarboxylic acid; [0181]
7-(4-{[(5-Cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)met-
hyl]oxy}phenyl)-3-isoquinolinecarboxylic acid; [0182]
6-[4-({[3-{[(2,6-Dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinoline carboxylic acid; [0183]
6-(4-{[(5-(1-Methylethyl)-3-{[(2,4,6-trifluorophenyl)oxy]methyl}-4-isoxaz-
olyl)methyl]oxy}phenyl)-2-quinoline carboxylic acid; [0184]
6-[4-({[3-{[(2,6-Dichlorophenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinoline carboxylic acid; [0185]
6-[4-({[3-{[(2,6-Dichlorophenyl)amino]methyl}-5-(1-methylethyl)-4-isoxazo-
lyl]methyl}oxy)phenyl]-2-quinoline carboxylic acid; [0186]
6-[4-({[3-{[(2,6-Dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-ethyl}ox-
y [0187]
6-[4-({[3-{[(2,6-Dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl-
)-4-isoxazolyl]methyl}oxy)phenyl]-2-quinoline carboxylic acid;
[0188]
6-[4-({[3-{[(2,6-Dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinoline carboxylic acid; [0189]
6-[4-({[3-(3,5-Dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid; [0190]
6-(4-{[(3,5-Dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinolinecarb-
oxylic acid [0191]
6-[4-({[3-[(Cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylic acid; [0192]
6-[4-({[3-(2,4-Dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid; [0193]
6-[4-({[3-(2,4-Dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid; [0194]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-7-fluoro-2-quinolinecarboxylic acid; [0195]
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-methyl-2-quinolinecarboxylic acid; [0196]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-3-methylphenyl]-2-quinolinecarboxylic acid; [0197]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-2-methylphenyl]-2-quinolinecarboxylic acid; [0198]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-methyl-2-quinolinecarboxylic acid; [0199]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-methyl-3-quinolinecarboxylic acid; [0200]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-1-isoquinolinecarboxylic acid; [0201]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-methyl-2-quinazolinecarboxylic acid; [0202]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio-
)phenyl]-2-quinolinecarboxylic acid; [0203]
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinoxalinecarboxylic acid; [0204] and pharmaceutically
acceptable salts or solvates thereof.
[0205] One preferred compound of the invention is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid; or a pharmaceutically
acceptable salt or solvate thereof. In one particular embodiment,
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid; or pharmaceutically acceptable
salt or solvate thereof is in crystalline form. In one preferred
embodiment, the compound of the invention is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid (i.e. the form of the acid).
[0206] One preferred compound of the invention is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid; or a pharmaceutically
acceptable salt or solvate thereof. In one particular embodiment,
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid; or pharmaceutically
acceptable salt or solvate thereof is in crystalline form. In one
preferred embodiment, the compound of the invention is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid (i.e. the form of the
acid).
[0207] Certain compounds of formula (I) may exist in stereoisomeric
forms (e.g. they may contain one or more asymmetric carbon atoms).
The individual stereoisomers (enantiomers and diastereomers) and
mixtures of these are included within the scope of the present
invention. The present invention also covers the individual isomers
of the compounds represented by formula (I) as mixtures with
isomers thereof in which one or more chiral centers are
inverted.
[0208] Suitable pharmaceutically acceptable salts according to the
present invention will be readily determined by one skilled in the
art and will include, for example, salts prepared from inorganic
bases such as lithium hydroxide, sodium hydroxide, potassium
hydroxide, lithium hydride, sodium hydride, potassium hydride,
lithium carbonate, lithium hydrogen carbonate, sodium carbonate,
sodium hydrogen carbonate, potassium carbonate, potassium hydrogen
carbonate, as well as potassium tert-butoxide and organic bases
such as diethyl amine, lysine, arginine, choline,
tris(hydroxymethyl)aminomethane(tromethamine), triethanolamine,
diethanolamine, and ethanolamine. In one embodiment, the compounds
of formula (I) are in the form of the potassium salt.
[0209] When used in medicine, the salts of a compound of formula
(I) should be pharmaceutically acceptable, but pharmaceutically
unacceptable salts may conveniently be used to prepare the
corresponding free base or pharmaceutically acceptable salts
thereof. One specific example of a salt of a compound of the
invention is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid potassium salt.
[0210] As used herein, the term "solvate" refers to a crystal form
containing the compound of formula (I) or a pharmaceutically
acceptable salt thereof and either a stoichiometric or a
non-stoichiometric amount of a solvent. Solvents, by way of
example, include water (thus producing hydrates), methanol,
ethanol, or acetic acid. Hereinafter, reference to a compound of
formula (I) is to any physical form of that compound, unless a
particular form, salt or solvate thereof is specified.
[0211] Processes for preparing pharmaceutically acceptable salts
and solvates of the compounds of formula (I) are conventional in
the art. See, e.g., Burger's Medicinal Chemistry And Drug Discovery
5th Edition, Vol 1: Principles And Practice.
[0212] As will be apparent to those skilled in the art, in the
processes described below for the preparation of compounds of
formula (I), certain intermediates, may be in the form of
pharmaceutically acceptable salts and/or solvates of the compound.
Those terms as applied to any intermediate employed in the process
of preparing compounds of formula (I) have the same meanings as
noted above with respect to compounds of formula (I). Processes for
preparing pharmaceutically acceptable salts and solvates of such
intermediates are known in the art and are analogous to the process
for preparing pharmaceutically acceptable salts and solvates of the
compounds of formula (I).
[0213] In one embodiment, the compounds of formula (I) are FXR
agonists. As used herein, the term "FXR agonist" refers to
compounds which exhibit a pEC.sub.50 greater than 4 in the FXR
Cofactor Recruitment Assay described below. More particularly, FXR
agonists are compounds which exhibit a pEC.sub.50 greater than 5 in
the FXR Cofactor Recruitment Assay described below.
[0214] Compounds of formula (I) are useful in therapy in subjects
such as mammals, and particularly humans. In particular, the
compounds of formula (I) are useful in the treatment of a condition
mediated by decreased FXR activity in a subject such as a mammal,
particularly a human. As used herein, the term "treatment" includes
the prevention of occurrence of symptoms of the condition or
disease in the subject, the prevention of recurrence of symptoms of
the condition or disease in the subject, the delay of recurrence of
symptoms of the condition or disease in the subject, the decrease
in severity or frequency of outward symptoms of the condition or
disease in the subject, slowing or eliminating the progression of
the condition and the partial or total elimination of symptoms of
the disease or condition in the subject.
[0215] Conditions which have been reported to be mediated by a
decreased FXR activity include but are not limited to dyslipidemia
(Sinal, C., et al. 2000 Cell 102:731-744; Zhang, Y., et al., 2006
Proc. Nat. Acad. Sci., U.S.A., 103:1006-1011); cardiovascular
diseases such as atherosclerosis (Hanniman, E. A., et al., J. Lipid
Res. 2005, 46:2595-2604); obesity (Chen, L., et al., 2006 Diabetes
55 suppl. 1:A200; Cariou, B., et al., 2006 J. Biol. Chem.
281:11039-11049; Rizzo, G., et al. 2006 Mol. Pharmacol.
70:1164-1173); diabetes mellitus (Duran-Sandoval, D., et al., 2004
Diabetes 53:890-898; Bilz, S., et al., 2006 Am. J. Physiol.
Endocrinol. Metab. 290:E716-E722; Nozawa, H., 2005 Biochem.
Biophys. Res. Commun. 336:754-761; Duran-Sandoval, D., et al., 2005
Biochimie 87:93-98; Claudel, T., et al., 2005 Arterioscler. Thromb.
Vasc. Biol. 25:2020-2030; Duran-Sandoval, D., et al., 2005 J. Biol.
Chem. 280:29971-29979; Savkur, R. S., et al., 2005 Biochem.
Biophys. Res. Commun., 329:391-396; Cariou, B., et al., 2006 J.
Biol. Chem. 281:11039-11049; Ma, K., et al., 2006 J. Clin. Invest.
116:1102-1109; Zhang, Y., et al., 2006 Proc. Nat. Acad. Sci. U.S.A.
103:1006-1011); metabolic syndrome (Chen, L., et al., 2006 Diabetes
55 suppl. 1:A200); disorders of the liver such as cholestatic liver
disease (Liu, Y. et al., 2003 J. Clin. Invest. 112:1678-1687) and
cholesterol gallstone disease (Moschetta, A., et al., 2004 Nat.
Med. 10:1352-1358); organ fibrosis (Fiorucci, S., et al. 2004
Gastroenterology 127:1497-1512 and Fiorucci, S., et al., 2005 J.
Pharmacol. Exp. Ther. 314:584-595) including liver fibrosis
(Fiorucci, S., et al. 2004 Gastroenterology 127:1497-1512);
inflammatory bowel disease (Inagaki, T., et al., 2006 Proc. Nat.
Acad. Sci., U. S. A. 103:3920-3925); and liver regeneration (Huang,
W., et al., 2006 Science 312:233-236).
[0216] Compounds of formula (I) are believed to be useful for the
treatment of dyslipidemia in a subject, such as a mammal,
particularly a human. The compounds of the present invention are
currently believed to increase the flow of bile acid. Increased
flow of bile acids improves the flux of bile acids from the liver
to the intestine. FXR null mice demonstrate that FXR not only plays
a role in bile acid homeostasis, but also plays a role in lipid
homeostasis by virtue of the regulation of enzymes and transporters
that are involved in lipid catabolism and excretion.
[0217] Compounds of formula (I) are also believed to be useful for
lowering triglycerides in a subject, such as a mammal, particularly
a human. As used herein "lowering triglycerides" means lowering
triglycerides in a subject in need thereof below the initial level
of triglyercides in that subject before administration of a
compound of formula (I). For example, the compounds of formula (I)
may lower triglycerides by decreasing fat absorption, decreasing
hepatic triglyceride production or decreasing hepatic triglyceride
secretion. The compounds of formula (I) may also lower serum and
hepatic triglycerides.
[0218] By treating dyslipidemia, compounds of formula (I) are
currently believed to be useful in the treatment of
hypertriglyceridemia and hypercholesteronemia related
cardiovascular disease such as atherosclerosis in a subject such as
a mammal, particularly a human. Compounds of formula (I) are also
believed to be useful for the treatment of non-alcoholic fatty
liver disease and non-alcoholic steatohepatitis in a subject, such
as a mammal, particularly a human (Chen, L., et al., 2006 Diabetes
55 suppl. 1:A200; Watanabe, M., et al., 2004 J. Clin. Invest.,
113:1408-1418).
[0219] The compounds of formula (I) are useful for the treatment of
obesity in a subject, such as a mammal, particularly a human.
[0220] Compounds of formula (I) are also useful for the treatment
of diabetes mellitus in a subject, such as a mammal, particularly a
human. For example, the compounds of formula (I) are useful for the
treatment of type 2 diabetes. The effects of an FXR agonist,
GW4064, on body weight, glucose tolerance, serum glucose, serum
insulin, serum triglyceride, and liver triglyceride contents via
oral administration have been observed in an high-fat diet induced
insulin resistant, glucose intolerant, and obese mouse model (Chen,
L., et al., 2006 Diabetes 55 suppl. 1:A200). Male 20 to 25 g C57BL
mice (Charles River, Indianapolis, Ind.) were housed at 72.degree.
F. and 50% relative humidity with a 12 h light and dark cycle and
fed with standard rodent chow (Purina 5001, Harlan Teklad,
Indianapolis, Ind.) or a high-fat diet (TD93075, Harlan Teklad,
Indianapolis, Ind.) for seven weeks. After two weeks, mice on
high-fat diet were randomized to vehicle or treatment groups. There
were no significant difference in body weight, body fat mass, serum
glucose and insulin, and area under the curve (AUC) for glucose in
glucose tolerance test (GTT) between the vehicle group and the
treatment group. Starting from the fourth week, mice were given
either vehicle or GW4064 (100 mg/kg) twice a day orally. Mice on
the standard rodent chow were also given vehicle as a control. At
the end of the third week of compound treatment, a GTT was
performed and body composition was measured using the quantitative
magnetic resonance (QMR) method. At the end of the study (fourth
week of compound treatment), blood samples were taken from inferior
vena cava and tissue samples were collected for further analysis.
Blood glucose during GTT was measured using Bayer Glucometer
Elite.RTM. XL. Serum chemistry levels were measured using the
Instrumentation Laboratory Ilab600.TM. clinical chemistry analyzer
(Instrumentation Laboratory, Boston, Mass.). Liver triglyceride
contents were measured using the methanolic-KOH saponification
method and a triglyceride assay kit (GPO-TRINDER, Sigma
Diagnostics, St. Louis, Mo.). The results indicated that GW4064
reduced the high-fat diet induced body weight gain. It is believed
that the result may have been due to a decrease in fat mass. GW4064
also appeared to improve glucose tolerance, decreased serum
glucose, insulin and triglyceride, and reduced liver triglyceride
content. In addition, Cariou and colleagues treated male ob/ob mice
with GW4064 (30 mg/kg) intraperitoneally (2006 J. Biol. Chem.
281:11039-11049). GW4064 treatment did not alter body weight as
well as food intake. Whereas GW4064 had no effect on fasting blood
glucose in ob/ob mice, it decreased insulin concentration in the
treated group. GW4064 treated ob/ob mice also showed an improved
glucose tolerance and enhanced insulin sensitivity compared to
controls. In another study, it was reported that GW4064
significantly improved hyperglycemia and hyperlipidemia in diabetic
db/db mice (Zhang, Y., et al, 2006 Proc. Nat. Acad. Sci. U.S.A.
103:1006-1011). Oral GW4064 (30 mg/kg, bid) treatment decreased
blood glucose, serum .beta.-hydroxybutyrate, triglyceride, NEFA,
and total cholesterol in db/db mice. It was also demonstrated that
GW4064 treatment enhanced insulin signalling and glycogen storage
in the liver of db/db mice. These data suggest that FXR agonists,
including the compounds of the formula (I), may be used for the
treatment of obesity, insulin resistance, glucose intolerance,
diabetes mellitus, fatty liver disease and metabolic syndrome.
[0221] Compounds of formula (I) are also useful for the treatment
of metabolic syndrome in a subject, such as a mammal, particularly
a human. Metabolic syndrome is characterized by a group of
metabolic risk factors in one person. They include abdominal
obesity (excessive fat tissue in and around the abdomen),
atherogenic dyslipidemia (high triglycerides, low high density
lipoprotein (HDL) cholesterol and high low density lipoprotein
(LDL) cholesterol), elevated blood pressure, insulin resistance or
glucose intolerance, prothrombotic state and proinflammatory state.
People with metabolic syndrome are at increased risk of coronary
heart disease and atherosclerosis-related diseases (e.g., stroke
and peripheral vascular disease) and type 2 diabetes mellitus.
There are several clinical criteria for metabolic syndromes
including ATP III, WHO, and AACE (American Association of Clinical
Endocrinologists) (see tables, for review see Grundy, S. M., et
al., 2004 Circulation 109:433-438). The present invention provides
a method for the treatment of metabolic syndrome characterized by
abdominal obesity, atherogenic dyslipidemia and insulin resistance
with or without glucose interance, and may benefit other components
of metabolic syndrome in a subject.
TABLE-US-00001 TABLE 1 ATP III Clinical Identification of the
Metabolic Syndrome Risk Factor Defining Level Abdominal obesity,
given as waist circumference*.sup..dagger. Men >102 cm (>40
in) Women >88 cm (>35 in) Triglycerides .gtoreq.150 mg/dL HDL
cholesterol Men <40 mg/dL Women <50 mg/dL Blood pressure
.gtoreq.130/.gtoreq.85 mmHg Fasting glucose .gtoreq.110
mg/dL.sup..dagger-dbl. *Overweight and obesity are associated with
insulin resistance and the metabolic syndrome. However, the
presence of abdominal obesity is more highly correlated with the
metabolic risk factors than is an elevated BMI. Therefore, the
simple measure of waist circumference is recommended to identify
the body weight component of the metabolic syndrome.
.sup..dagger.Some male patients can develop multiple metabolic risk
factors when the waist circumference is only marginally increased,
eg, 94 to 102 cm (37 to 39 in). Such patients may have a strong
genetic contribution to insulin resistance. They should benefit
from changes in life habits, similarly to men with categorical
increases in waist circumference. .sup..dagger-dbl.The American
Diabetes Association has recently established a cutpoint of
.gtoreq.100 mg/dL, above which persons have either prediabetes
(impaired fasting glucose) or diabetes. This new cutpoint should be
applicable for identifying the lower boundary to define an elevated
glucose as one criterion for the metabolic syndrome.
TABLE-US-00002 TABLE 2 WHO Clinical Criteria for Metabolic Syndrome
Insulin resistance, identified by 1 of the following: Type 2
diabetes Impaired fasting glucose Impaired glucose tolerance or for
those with normal fasting glucose levels (<110 mg/dL), glucose
uptake below the lowest quartile for background population under
investigation under hyperinsulinemic, euglycemic conditions Plus
any 2 of the following: Antihypertensive medication and/or high
blood pressure (.gtoreq.140 mmHg systolic or .gtoreq.90 mmHg
diastolic) Plasma triglycerides .gtoreq.150 mg/dL (.gtoreq.1.7
mmol/L) HDL cholesterol <35 mg/dL (<0.9 mmol/L) in men or
<39 mg/dL (1.0 mmol/L) in women BMI > 30 kg/m.sup.2 and/or
waist:hip ratio >0.9 in men, >0.85 in women Urinary albumin
excretion rate .gtoreq.20 .mu.g/min or albumin:creatinine ratio
.gtoreq.30 mg/g
TABLE-US-00003 TABLE 3 AACE Clinical Criteria for Diagnosis of the
Insulin Resistance Syndrome* Risk Factor Components Cutpoints for
Abnormality Overweight/obesity BMI .gtoreq. 25 kg/m.sup.2 Elevated
triglycerides .gtoreq.150 mg/dL (1.69 mmol/L) Low HDL cholesterol
Men <40 mg/dL (1.04 mmol/L) Women <50 mg/dL (1.29 mmol/L)
Elevated blood pressure .gtoreq.130/85 mmHg 2-Hour postglucose
>140 mg/dL challenge Fasting glucose Between 110 and 126 mg/dL
Other risk factors Family history of type 2 diabetes, hypertension,
or CVD Polycystic ovary syndrome Sedentary lifestyle Advancing age
Ethnic groups having high risk for type 2 diabetes or CVD
*Diagnosis depends on clinical judgment based on risk factors.
[0222] Compounds of formula (I) are believed to be useful for the
treatment of cholestatic liver disease. For example, the compounds
of formula (I) are believed to be useful in the treatment of
primary biliary cirrhosis or primary sclerosing cholangitis. FXR
therefore is a target for the treatment of a number of cholestatic
liver diseases and non-alcoholic steatohepatitis. The compounds of
formula (I) are also believed to be useful for the treatment of
gall stones. For example, the compounds of formula (I) are believed
to be useful in the treatment of cholesterol gallstone disease. The
compounds of formula (I) are also believed to be useful for
decreasing liver lipid accumulation.
[0223] Compounds of formula (I) are also believed to be useful for
the treatment of organ fibrosis. Fibrotic disorders can be
characterized as acute or chronic, but share the common
characteristic of excessive collagen accumulation and an associated
loss of function as normal tissues are replaced or displaced by
fibrotic tissues. Acute forms of fibrosis include response to
trauma, infections, surgery, burns, radiation and chemotherapy.
Chronic forms of fibrosis may be due to viral infection, diabetes
mellitus, obesity, fatty liver, hypertension, scleroderma and other
chronic conditions that induce fibrosis.
[0224] Organs that are most commonly affected by fibrosis include
liver, kidney, and lung. Organ fibrosis can cause the progressive
loss of organ function. Retroperitoneal fibrosis (including
idiopathic retroperitoneal fibrosis) may not originate from any
major organ, but can involve and adversely affect the function of
organs such as the kidneys.
[0225] Accordingly, as used herein, the term fibrosis refers to all
recognized fibrotic disorders, including fibrosis due to
pathological conditions or diseases, fibrosis due to physical
trauma (`traumatic fibrosis`), fibrosis due to radiation damage,
and fibrosis due to exposure to chemotherapeutics. As used herein,
the term "organ fibrosis" includes but is not limited to liver
fibrosis, fibrosis of the kidneys, fibrosis of lung, and fibrosis
of the intestine. "Traumatic fibrosis" includes but is not limited
to fibrosis secondary to surgery (surgical scarring), accidental
physical trauma, burns, and hypertrophic scarring.
[0226] In one embodiment, compounds of formula (I) are useful for
the treatment of liver fibrosis in a subject, particularly a mammal
such as a human, in need of treatment thereof. As used herein,
"liver fibrosis" includes liver fibrosis due to any cause,
including but not limited to virally-induced liver fibrosis such as
that due to hepatitis B or C virus; exposure to alcohol (alcoholic
liver disease), certain pharmaceutical compounds including but not
limited to methotrexate, some chemotherapeutic agents, and chronic
ingestion of arsenicals or vitamin A in megadoses, oxidative
stress, cancer radiation therapy or certain industrial chemicals
including but not limited to carbon tetrachloride and
dimethylnitrosamine; and diseases such as primary biliary
cirrhosis, primary sclerosing colangitis, fatty liver, obesity,
non-alcoholic steatohepatitis, cystic fibrosis, hemochromatosis,
auto-immune hepatitis, and steatohepatitis. Current therapy in
liver fibrosis is primarily directed at removing the causal agent,
e.g., removing excess iron (e.g., in the case of hemochromatosis),
decreasing viral load (e.g., in the case of chronic viral
hepatitis), or eliminating or decreasing exposure to toxins (e.g.,
in the case of alcoholic liver disease). Anti-inflammatory drugs
such as corticosteroids and colchicine are also known for use in
treating inflammation that can lead to liver fibrosis. Other
strategies for treating liver fibrosis are under development (see,
e.g., Murphy, F., et al., 2002 Expert Opin. Invest. Drugs
11:1575-1585; Bataller, R. and Brenner, D. A., 2001 Sem. Liver Dis.
21:437-451). Thus in another embodiment, the present invention
provides a method for the treatment of liver fibrosis in a subject
which comprises administering a therapeutically effective amount of
a compound of formula (I) in combination with another therapeutic
agent useful for the treatment of symptoms associated with liver
fibrosis. Examples of therapeutic agents useful for the treatment
of symptoms associated with liver fibrosis include corticosteroids
and cholchicine.
[0227] The response of the liver to hepatocellular damage, similar
to wound healing in other tissues, includes inflammation and tissue
remodeling, with associated changes in the quantity and quality of
the extracellular matrix. Progressive accumulation of extracellular
matrix proteins, including collagen types I and III, eventually
distorts the architecture of the liver by forming fibrous scars,
resulting in disrupted blood flow and an eventual deterioration in
hepatic function. (Bissell, D. M. and Maher, J. J., "Hepatic
Fibrosis and Cirrhosis." Ed. Zakim, D. and Thomas, D. B., 4 ed. 2
vols. Philadelphia: Saunders, 2003. 395-416, Hanauske-Abel, H. M.,
"Fibrosis of the Liver: Representative Molecular Elements and Their
Emerging Role As Anti-Fibrotic Targets." Ed. Zakim, D., and Thomas,
D. B., 4 ed. 2 vols. Philadelphia: Saunders, 2003. 347-394).
Hepatic stellate cells (HSC) have been identified as important
mediators of the fibrotic process in the liver, and are believed to
be primarily responsible for the synthesis of excess extracellular
matrix seen in liver disease. Liver injury can result in quiescent
HSCs converting to activated myofibroblast-like cells that
proliferate, migrate, recruit inflammatory cells, and synthesize
collagens and other extracellular matrix proteins. (Bissell, D. M.
and Maher, J. J., "Hepatic Fibrosis and Cirrhosis." Ed. Zakim, D.
and Thomas, D. B., 4 ed. 2 vols. Philadelphia: Saunders, 2003.
395-416, Hanauske-Abel, H. M., "Fibrosis of the Liver:
Representative Molecular Elements and Their Emerging Role As
Anti-Fibrotic Targets." Ed. Zakim, D., and Thomas, D. B., 4 ed. 2
vols. Philadelphia: Saunders, 2003. 347-394). Various cytokines are
reported to activate HSCs, including transforming growth factor
.beta. (TGF.beta.). Following liver injury, HSCs synthesize
.alpha.-smooth muscle actin (.alpha.-SMA) as part of the migration
response, consequently a marked accumulation of .alpha.-SMA can be
seen at areas of active liver fibrogenesis. (Bissell, D. M. and
Maher, J. J., "Hepatic Fibrosis and Cirrhosis." Ed. Zakim, D. and
Thomas, D. B., 4 ed. 2 vols. Philadelphia: Saunders, 2003. 395-416,
Hanauske-Abel, H. M., "Fibrosis of the Liver: Representative
Molecular Elements and Their Emerging Role As Anti-Fibrotic
Targets." Ed. Zakim, D., and Thomas, D. B., 4 ed. 2 vols.
Philadelphia: Saunders, 2003. 347-394). Derangement of the normal
epithelial/mesenchymal interaction, characterised by cholangiocyte
damage/proliferation, can also lead to extracellular
matrix-producing and progressive fibrogenesis. [Pinzani, M., et
al., 2004 Digest. Liver Dis. 36:231-242.]
[0228] As is known in the art, liver fibrosis may be clinically
classified into five stages of severity (S0 to S4), usually based
on histological examination of a biopsy specimen. S0 indicates no
fibrosis, whereas S4 indicates cirrhosis. While various criteria
for staging the severity of liver fibrosis exist, in general early
stages of fibrosis are identified by discrete, localized areas of
scarring in one portal (zone) of the liver, whereas later stages of
fibrosis are identified by bridging fibrosis (scarring that crosses
zones of the liver).
[0229] Compounds of formula (I) are also useful for the treatment
of inflammatory bowel disease in a subject, such as a mammal,
particularly a human. Inflammatory bowel disease (IBD) is defined
as a group of idiopathic relapsing inflammatory disorders of the
bowel--the large or small intestine. The pathogenesis of IBD
remains obscure and may involve genetic, environmental and
immunological factors. [Drossman, D. A. 1999 Aliment Pharmacol.
Ther. 13(s2):3-14; Danese, S., et al. 2004 Autoimmunity Reviews 3:
394-400; Stokkers, P. C. F. and Hommes, D. W. 2004 Cytokine
28:167-173.] The most common types of inflammatory bowel disease
are ulcerative colitis and Crohn disease.
[0230] Compounds of formula (I) are also believed to be useful for
enhancing liver regeneration in a subject, such as a mammal,
particularly a human. For example, the compounds of formula (I) are
believed to be useful for enhancing liver regeneration for liver
transplantation.
[0231] The present invention provides a method for the treatment of
a condition mediated by decreased FXR activity, particularly a
condition in which a FXR agonist may be useful, in a subject, such
as a mammal, particularly a human, in need thereof. The present
invention also provides the use of a compound of formula (I) for
the preparation of a medicament for the treatment of a condition
mediated by decreased FXR activity, particularly a condition in
which a FXR agonist may be useful, in a subject, such as a mammal,
particularly a human in need thereof.
[0232] The present invention also provides a method for lowering
triglycerides in a subject, such as a mammal, particularly a human,
in need thereof. The present invention also provides the use of a
compound of formula (I) for the preparation of a medicament for
lowering triglycerides in a subject. In one embodiment, the
compound of formula (I) is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]meth-
yl}oxy)phenyl]-2-quinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof. In another embodiment, the compound of
formula (I) is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof.
[0233] The present invention provides a method for the treatment of
obesity in a subject, such as a mammal, particularly a human, in
need thereof. The present invention also provides the use of a
compound of formula (I) for the preparation of a medicament for the
treatment of obesity in a subject. In one embodiment, the compound
of formula (I) is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid or a pharmaceutically acceptable
salt thereof. In another embodiment, the compound of formula (I) is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof.
[0234] The present invention provides a method for the treatment of
diabetes mellitus in a subject, such as a mammal, particularly a
human, in need thereof. The present invention also provides the use
of a compound of formula (I) for the preparation of a medicament
for the treatment of diabetes mellitus in a subject. In one
embodiment, the compound of formula (I) is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid or a pharmaceutically acceptable
salt thereof. In another embodiment, the compound of formula (I) is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof.
[0235] The present invention provides a method for the treatment of
metabolic syndrome in a subject, such as a mammal, particularly a
human, in need thereof. The present invention also provides the use
of a compound of formula (I) for the preparation of a medicament
for the treatment of metabolic syndrome in a subject. In one
embodiment, the compound of formula (I) is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid or a pharmaceutically acceptable
salt thereof. In another embodiment, the compound of formula (I) is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof.
[0236] The present invention provides a method for the treatment of
cholestatic liver disease in a subject, such as a mammal,
particularly a human, in need thereof. The present invention also
provides the use of a compound of formula (I) for the preparation
of a medicament for the treatment of cholestatic liver disease in a
subject. In one embodiment, the compound of formula (I) is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid or a pharmaceutically acceptable
salt thereof. In another embodiment, the compound of formula (I) is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof.
[0237] The present invention provides a method for the treatment of
organ fibrosis in a subject, such as a mammal, particularly a
human, in need thereof. The present invention also provides the use
of a compound of formula (I) for the preparation of a medicament
for the treatment of organ fibrosis in a subject. In one
embodiment, the compound of formula (I) is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]meth-
yl}oxy)phenyl]-2-quinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof. In another embodiment, the compound of
formula (I) is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof.
[0238] The present invention provides a method for the treatment of
liver fibrosis in a subject, such as a mammal, particularly a
human, in need thereof. The present invention also provides the use
of a compound of formula (I) for the preparation of a medicament
for the treatment of liver fibrosis in a subject. In one
embodiment, the compound of formula (I) is
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]meth-
yl}oxy)phenyl]-2-quinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof. In another embodiment, the compound of
formula (I) is
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid or a pharmaceutically
acceptable salt thereof.
[0239] All of the methods of the present invention comprise the
step of administering a therapeutically effective amount of the
compound of formula (I) or a pharmaceutically acceptable salt or
solvate thereof. As used herein, the term "therapeutically
effective amount" refers to an amount of a compound of formula (I)
which is sufficient to achieve the stated effect in the subject to
which it is administered. Accordingly, a therapeutically effective
amount of a compound of formula (I) used in the method for the
treatment of a condition mediated by decreased FXR activity in a
human will be an amount sufficient for the treatment of the
condition mediated by decreased FXR activity in a human. A
therapeutically effective amount of a compound of formula (I) for
use in the method for the treatment of diabetes mellitus in a human
will be an amount sufficient for the treatment of diabetes mellitus
in a human. A therapeutically effective amount of a compound of
formula (I) for use in the method for the treatment of metabolic
syndrome in a human will be an amount sufficient for the treatment
of metabolic syndrome in a human. A therapeutically effective
amount of a compound of formula (I) for use in the method for the
treatment of organ (e.g., liver) fibrosis in a human will be an
amount sufficient for the treatment of organ fibrosis in a
human.
[0240] The amount of a compound of formula (I) which is required to
achieve the desired therapeutic or biological effect will depend on
a number of factors such as the use for which it is intended, the
means of administration, the recipient and the type and severity of
the condition or disease being treated, and will be ultimately at
the discretion of the attendant physician or veterinarian. In
general, a typical daily dose for the treatment of a disease or
condition mediated by decreased FXR activity in a human, for
instance, may be expected to lie in the range of from about 0.01
mg/kg to about 100 mg/kg for a 70 kg human. This dose may be
administered as a single unit dose or as several separate unit
doses or as a continuous infusion. Similar dosages would be
applicable for the treatment of other diseases, conditions and
therapies including diabetes mellitus and obesity in humans.
[0241] While it is possible that, for use in therapy, a
therapeutically effective amount of a compound of formula (I) may
be administered as the raw chemical, it is typically presented as
the active ingredient of a pharmaceutical composition or
formulation. Accordingly, the invention further provides a
pharmaceutical composition comprising a compound of the formula
(I). The pharmaceutical composition may further comprise one or
more pharmaceutically acceptable carriers or diluents. The
carrier(s) and/or diluent(s) must be acceptable in the sense of
being compatible with the other ingredients of the formulation and
not deleterious to the recipient thereof. In one particular
embodiment, the compound is in crystalline form. The invention also
specifically contemplates compositions comprising the potassium
salt of the aforementioned compound. In accordance with another
aspect of the invention there is also provided a process for the
preparation of a pharmaceutical formulation including admixing a
compound of the formula (I) with one or more pharmaceutically
acceptable carriers and/or diluents.
[0242] Pharmaceutical formulations may be presented in unit dose
form containing a predetermined amount of active ingredient per
unit dose. Such a unit may contain a therapeutically effective dose
of the compound of formula (I) or a fraction of a therapeutically
effective dose such that multiple unit dosage forms might be
administered at a given time to achieve the desired therapeutically
effective dose. Preferred unit dosage formulations are those
containing a daily dose or sub-dose, as herein above recited, or an
appropriate fraction thereof, of an active ingredient. Furthermore,
such pharmaceutical formulations may be prepared by any of the
methods well known in the pharmacy art.
[0243] Pharmaceutical formulations may be adapted for
administration by any appropriate route, for example by the oral
(including buccal or sublingual), rectal, nasal, topical (including
buccal, sublingual or transdermal), vaginal or parenteral
(including subcutaneous, intramuscular, intravenous or intradermal)
route. Such formulations may be prepared by any method known in the
art of pharmacy, for example by bringing into association the
active ingredient with the carrier(s) or excipient(s).
[0244] Pharmaceutical formulations adapted for oral administration
may be presented as discrete units such as capsules or tablets;
powders or granules; solutions or suspensions in aqueous or
non-aqueous liquids; edible foams or whips; or oil-in-water liquid
emulsions or water-in-oil liquid emulsions.
[0245] For instance, for oral administration in the form of a
tablet or capsule, the active drug component can be combined with
an oral, non-toxic pharmaceutically acceptable inert carrier such
as ethanol, glycerol, water and the like. Powders are prepared by
comminuting the compound to a suitable fine size and mixing with a
similarly comminuted pharmaceutical carrier such as an edible
carbohydrate, as, for example, starch or mannitol. Flavoring,
preservative, dispersing and coloring agent can also be
present.
[0246] Capsules are made by preparing a powder mixture as described
above, and filling formed gelatin sheaths. Glidants and lubricants
such as colloidal silica, talc, magnesium stearate, calcium
stearate or solid polyethylene glycol can be added to the powder
mixture before the filling operation. A disintegrating or
solubilizing agent such as agar-agar, calcium carbonate or sodium
carbonate can also be added to improve the availability of the
medicament when the capsule is ingested.
[0247] Moreover, when desired or necessary, suitable binders,
lubricants, disintegrating agents and coloring agents can also be
incorporated into the mixture. Suitable binders include starch,
gelatin, natural sugars such as glucose or beta-lactose, corn
sweeteners, natural and synthetic gums such as acacia, tragacanth
or sodium alginate, carboxymethyl-cellulose, polyethylene glycol,
waxes and the like. Lubricants used in these dosage forms include
sodium oleate, sodium stearate, magnesium stearate, sodium
benzoate, sodium acetate, sodium chloride and the like.
Disintegrators include, without limitation, starch, methyl
cellulose, agar, bentonite, xanthan gum and the like. Tablets are
formulated, for example, by preparing a powder mixture, granulating
or slugging, adding a lubricant and disintegrant and pressing into
tablets. A powder mixture is prepared by mixing the compound,
suitably comminuted, with a diluent or base as described above, and
optionally, with a binder such as carboxymethylcellulose, an
aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant
such as paraffin, a resorption accelerator such as a quaternary
salt and/or an absorption agent such as bentonite, kaolin or
dicalcium phosphate. The powder mixture can be granulated by
wetting with a binder such as syrup, starch paste, acadia mucilage
or solutions of cellulosic or polymeric materials and forcing
through a screen. As an alternative to granulating, the powder
mixture can be run through the tablet machine and the result is
imperfectly formed slugs broken into granules. The granules can be
lubricated to prevent sticking to the tablet forming dies by means
of the addition of stearic acid, a stearate salt, talc or mineral
oil. The lubricated mixture is then compressed into tablets. The
compounds of the present invention can also be combined with a free
flowing inert carrier and compressed into tablets directly without
going through the granulating or slugging steps. A clear or opaque
protective coating consisting of a sealing coat of shellac, a
coating of sugar or polymeric material and a polish coating of wax
can be provided. Dyestuffs can be added to these coatings to
distinguish different unit dosages.
[0248] Oral fluids such as solution, syrups and elixirs can be
prepared in dosage unit form so that a given quantity contains a
predetermined amount of active ingredient. Syrups can be prepared
by dissolving the compound in a suitably flavored aqueous solution,
while elixirs are prepared through the use of a non-toxic alcoholic
vehicle. Suspensions can be formulated by dispersing the compound
in a non-toxic vehicle. Solubilizers and emulsifiers such as
ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol
ethers, preservatives, flavor additive such as peppermint oil or
natural sweeteners or saccharin or other artificial sweeteners, and
the like can also be added.
[0249] Where appropriate, dosage unit formulations for oral
administration can be microencapsulated. The formulation can also
be prepared to prolong or sustain the release as for example by
coating or embedding particulate material in polymers, wax or the
like.
[0250] A compound of formula (I) can also be administered in the
form of liposome delivery systems, such as small unilamellar
vesicles, large unilamellar vesicles and multilamellar vesicles.
Liposomes can be formed from a variety of phospholipids, such as
cholesterol, stearylamine or phosphatidylcholines.
[0251] A compound of formula (I) may also be delivered by the use
of monoclonal antibodies as individual carriers to which the
compound molecules are coupled. The compounds may also be coupled
with soluble polymers as targetable drug carriers. Such polymers
can include polyvinylpyrrolidone, pyran copolymer,
polyhydroxypropylmethacrylamide-phenol,
polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine
substituted with palmitoyl residues. Furthermore, the compounds may
be coupled to a class of biodegradable polymers useful in achieving
controlled release of a drug, for example, polylactic acid,
polyepsilon caprolactone, polyhydroxy butyric acid,
polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates
and cross-linked or amphipathic block copolymers of hydrogels.
[0252] Pharmaceutical compositions adapted for transdermal
administration may be presented as discrete patches intended to
remain in intimate contact with the epidermis of the recipient for
a prolonged period of time. For example, the active ingredient may
be delivered from the patch by iontophoresis as generally described
in 1986 Pharmaceutical Research 3:318.
[0253] Pharmaceutical compositions adapted for topical
administration may be formulated as ointments, creams, suspensions,
lotions, powders, solutions, pastes, gels, sprays, aerosols or
oils.
[0254] For treatments of the eye or other external tissues, for
example mouth and skin, the compositions are preferably applied as
a topical ointment or cream. When formulated in an ointment, the
active ingredient may be employed with either a paraffinic or a
water-miscible ointment base. Alternatively, the active ingredient
may be formulated in a cream with an oil-in-water cream base or a
water-in-oil base.
[0255] Pharmaceutical compositions adapted for topical
administrations to the eye include eye drops wherein the active
ingredient is dissolved or suspended in a suitable carrier,
especially an aqueous solvent.
[0256] Pharmaceutical compositions adapted for topical
administration in the mouth include lozenges, pastilles and mouth
washes.
[0257] Pharmaceutical compositions adapted for rectal
administration may be presented as suppositories or as enemas.
[0258] Pharmaceutical compositions adapted for nasal administration
wherein the carrier is a solid include a coarse powder having a
particle size for example in the range of about 20 microns to about
500 microns which is administered in the manner in which snuff is
taken, i.e. by rapid inhalation through the nasal passage from a
container of the powder held close up to the nose. Suitable
formulations wherein the carrier is a liquid, for administration as
a nasal spray or as nasal drops, include aqueous or oil solutions
of the active ingredient.
[0259] Pharmaceutical compositions adapted for administration by
inhalation include fine particle dusts or mists, which may be
generated by means of various types of metered, dose pressurised
aerosols, nebulizers or insufflators.
[0260] Pharmaceutical compositions adapted for vaginal
administration may be presented as pessaries, tampons, creams,
gels, pastes, foams or spray formulations.
[0261] Pharmaceutical compositions adapted for parenteral
administration include aqueous and non-aqueous sterile injection
solutions which may contain anti-oxidants, buffers, bacteriostats
and solutes which render the formulation isotonic with the blood of
the intended recipient; and aqueous and non-aqueous sterile
suspensions which may include suspending agents and thickening
agents. The compositions may be presented in unit-dose or
multi-dose containers, for example sealed ampoules and vials, and
may be stored in a freeze-dried (lyophilized) condition requiring
only the addition of the sterile liquid carrier, for example water
for injections, immediately prior to use. Extemporaneous injection
solutions and suspensions may be prepared from sterile powders,
granules and tablets.
[0262] It should be understood that in addition to the ingredients
particularly mentioned above, the compositions may include other
agents conventional in the art having regard to the type of
formulation in question, for example those suitable for oral
administration may include flavouring agents.
[0263] In the above-described methods of treatment and uses, a
compound of formula (I) may be employed alone, in combination with
one or more other compounds of formula (I) or in combination with
other therapeutic agents. Thus, the present invention also
encompasses pharmaceutical compositions further comprising one or
more therapeutic agents. In one embodiment, the pharmaceutical
compositions further comprise one or more lipid-altering agents.
Examples of lipid-altering agents include but are not limited to
liver X receptor (LXR) agonists described in PCT Publication No.
WO02/24632 to GlaxoSmithKline.
[0264] Examples of other therapeutic agents include, but are not
limited to, 3-Hydroxy-3-Methyl-Glutaryl-CoA reductase inhibitors
such as statins (atorvastatin, fluvastatin, pravastatin,
lovastatin, cerivastatin, and nisvastatin); squalene epoxidase
inhibitors, squalene synthetase inhibitors, bile acid transport
inhibitors (BATi), human peroxisome proliferator activated receptor
(PPAR) gamma agonists such as rosiglitazone, troglitazone, and
pioglitazone and thiazolidinediones; PPAR .alpha. agonists such as
clofibrate, fenofibrate and gemfibronzil; PPAR dual .alpha./.gamma.
agonists; cyclooxygenase-2 (COX-2) inhibitors such as rofecoxib and
celecoxib; thrombin inhibitors; acyl-coenzyme A; cholesterol
acyltransferase (ACAT) inhibitors including selective ACAT
inhibitors; microsomal triglyceride transfer protein (MTP)
inhibitors; probucol, niacin; cholesterol absorption inhibitors;
bile acid sequestrants; LDL receptor inducers; platelet aggregation
inhibitors such as glycoprotein IIb/IIIa fibrinogen receptor
antagonists and aspirin; vitamin B6 and pharmaceutically acceptable
salts thereof; vitamin B12; folic acid or a pharmaceutically
acceptable salt or ester thereof; antioxidant vitamins such as C
and E and beta carotene; beta blockers; angiotensin II antagonists
such as losartan; antiotensin converting enzyme inhibitors such as
enalapril and captopril; calcium channel blockers such as
nifedipine and diltiazam; endothelian antagonists; agents other
than LXR ligands that enhance ATP-Binding Cassette Transporter-A1
gene expression; and bisphosphonate compounds such as alendronate
sodium.
[0265] The methods and uses employing these combinations may
comprise the administration of the compound of formula (I) and
another therapeutic agent either sequentially in any order or
simultaneously in separate or combined pharmaceutical compositions.
When combined in the same composition it will be appreciated that
the compounds must be stable and compatible with each other and the
other components of the composition and may be formulated for
administration. When formulated separately they may be provided in
any convenient formulation, in such a manner as are known for such
compounds in the art.
[0266] When a compound of formula (I) is used in combination with
another therapeutic agent, the dose of each compound may differ
from that when the compound is used alone. Appropriate doses will
be readily appreciated by those skilled in the art. The appropriate
dose of the compound(s) of formula (I) and the other
therapeutically active agent(s) and the relative timings of
administration will be selected in order to achieve the desired
combined therapeutic effect, and are within the expertise and
discretion of the attendant clinician.
[0267] Compounds of the invention can be made according to any
suitable method of organic chemistry. As will be apparent to those
skilled in the art and as depicted in the schemes which follow, the
order of the steps in each reaction is not critical to the practice
of the processes of the present invention. The reaction steps
depicted in each scheme may be carried out in any suitable order
based upon the knowledge of those skilled in the art. Further, it
will be apparent to those skilled in the art that certain reaction
steps may be most efficiently performed by installing protecting
groups prior to the reaction, which are removed subsequently. The
choice of protecting groups as well as general techniques for their
installation and removal are within the skill of those in the
art.
[0268] According to one method, a compound of formula (I) may be
prepared using the process depicted in Scheme 1, below.
##STR00036## [0269] wherein: [0270] X.sup.1 is chloride, iodide,
bromide, triflate, tosylate, nosylate, besylate or mesylate,
(preferably chloro); [0271] each R.sup.1 is the same or different
and is independently selected from alkyl, fluoroalkyl or
--CO.sub.2alkyl; wherein at least one R.sup.1 is --CO.sub.2alkyl;
and [0272] all other variables are as defined above for formula
(I).
[0273] In general, the process for preparing a compound of formula
(I) as depicted in Scheme 1 comprises the steps of: [0274] a)
reacting a compound of formula (II) with a compound of formula
(III) to prepare a compound of formula (I); [0275] d) optionally
converting the compound of formula (I) into a pharmaceutically
acceptable salt or solvate thereof; and [0276] e) optionally
converting the compound of formula (I) or a pharmaceutically
acceptable salt or solvate thereof into a different compound of
formula (I) or a pharmaceutically acceptable salt or solvate
thereof.
[0277] A compound of formula (I), prepared by any suitable process,
may be converted into a pharmaceutically acceptable salt or solvate
thereof or may be converted to a different compound of formula (I)
or a pharmaceutically acceptable salt or solvate thereof using
techniques described herein below and those conventional in the
art.
[0278] More particularly, the compound of formula (I) may be
prepared by reacting the compound of formula (II) with a compound
of formula (III) in the presence of a suitable base such as cesium
carbonate or potassium carbonate, in a polar aprotic solvent, such
as N,N-dimethylformamide, at ambient or elevated temperature.
[0279] The compound of formula (III) may be prepared by reacting a
compound of formula (IV) with the appropriate reagent to prepare a
compound having the desired leaving group (X.sup.1).
##STR00037## [0280] wherein all variables are as defined above.
[0281] In the embodiment wherein X.sup.1 is halide, the reaction is
performed by halogenating the compound of formula (IV). Any
suitable halogenating reagent conventional in the art may be
employed in the reaction. Examples of suitable halogenating
reagents include, but are not limited to, thionyl chloride and
triphenylphosphine dichloride. The reaction is typically carried
out in a non-polar solvent such as dichloromethane or
1,2-dichloroethane at ambient temperature.
[0282] In the embodiment wherein X.sup.1 is triflate, tosylate or
meslyate, the reaction process may be carried out according to the
conventional methods. See, Vedejs, E., et al., 1977 J. Org. Chem.
42:3109-3113; Handy, S. T., et al. 2004 J. Org. Chem. 69:2362-2366;
and Copp, F. C., et al. 1955 J. Chem. Soc. 2021-2027.
[0283] The compound of formula (IV) may be prepared by reducing a
compound of formula (V).
##STR00038## [0284] wherein all variables are as defined above.
[0285] A compound of formula (V) may be treated with a reducing
agent, such as diisobutylaluminum hydride, in a suitable solvent
such as tetrahydrofuran.
[0286] In another embodiment, the compound of formula (V) may be
saponified to the corresponding carboxylic acid prior to reducing
with a suitable reducing agent, such as borane, to prepare a
compound of formula (IV). In addition, the carboxylic acid may also
converted to a mixed anhydride before reducing with a reducing
agent such as sodium borohydride to prepare a compound of formula
(IV).
[0287] Compounds of formula (V) may be prepared by multiple routes.
In one embodiment, the compound of formula (V) may be prepared by a
process comprising the steps of: [0288] a) chlorinating a compound
of formula (VI); and [0289] b) cyclizing with a .beta.-ketoester of
formula (VII).
[0289] ##STR00039## [0290] wherein all variables are as defined
above.
[0291] The process is conveniently carried out according to the
method described by Doyle, F. P., et. al., 1963 J. Chem. Soc.
5838-5845. Esters of formula (VII) are commercially available or
can be prepared using conventional techniques.
[0292] The compound of formula (VI) may be prepared by condensing a
compound of formula (VIII) with hydroxylamine.
##STR00040## [0293] wherein all variables are as defined above.
[0294] Conditions suitable for this condensation reaction are
conventional in the art.
[0295] In another embodiment, a compound of formula (V) is prepared
by a process comprising the steps of: a) reacting a compound of
formula (IX) with tin chloride in the presence of a compound of
formula (VII) to prepare a compound of formula (X) and b) reacting
the compound of formula (X) with hydroxylamine to yield a compound
of formula (V). See, Singh, B. and Lesher, G. Y. 1978 Synthesis
829-830.
##STR00041## [0296] wherein all variables are as defined above.
[0297] The compound of formula (IX) may be obtained commercially or
by procedures in the literature. See, Guo, H. and Zhang, Y. 2000
Syn. Commun. 30:1879-1885. The compound of formula (V) may then be
reduced with a suitable reducing agent, such as diisobutylaluminum
hydride, in the manner described above, to prepare a compound of
formula (IV).
[0298] In another embodiment, a compound of formula (V) wherein d
is 1, may be prepared by a process comprising the steps of: a)
hydrolyzing a compound of formula (XI) to prepare a compound of
formula (XII) and b) reacting the alcohol of formula (XII) under
Mitsunobu conditions with a compound of formula (i-b), (ii-b) or
(iii-b).
##STR00042## [0299] wherein: [0300] c and d are each 1; [0301]
Z.sup.2 is --O--, --S-- or --NH--; [0302] Ring D is a moiety of
formula i, ii or iii; [0303] and all other variables are defined as
above.
[0304] The hydrolysis of the compound of formula (XI) may be
carried out using conventional techniques, including by reaction
with an acid, such as trifluoroacetic acid, in a solvent such as
dichloromethane or 1,2-dichloroethane. The resulting compound of
formula (XII) may be reacted under Mitsunobu reaction conditions
with a compound of formula of i, ii, or iii to prepare a compound
of formula (V). Suitable reaction conditions for the Mitsunobu
reaction are known to those skilled in the art. For example, the
reaction may be carried out in a solution of dichloromethane,
1,2-dichloroethane or toluene with triphenyl phosphine and a
dialkyl azodicarboxylate, such as diisopropyl azodicarboxylate or
di-tent-butyl azodicarboxylate. To prepare Compounds of formula (V)
wherein d is 1 and Z.sup.2 is NH it may be desirable to first
prepare the trifluoroacetamide derivative of the compound of
formula (i, ii, or iii, such that Z.sup.2 is NC(O)CF.sub.3) before
performing the Mitsunobu reaction. To prepare a compound of formula
(V) wherein Z.sup.2 is SO or SO.sub.2, a compound of formula (V)
wherein Z.sup.2 is S is oxidized using an oxidant, such as
3-chloroperbenzoic acid.
[0305] A compound of formula (V) may also be prepared by reacting
the compound of a formula (XII) with an appropriate reagent to
install a leaving group, such as chloro, iodo, bromo, triflate,
tosylate, nosylate, besylate or mesylate, (preferably bromo), to
prepare a compound of formula (XIII) having the desired leaving
group (X.sup.1).
##STR00043## [0306] wherein: [0307] X.sup.1 is selected from Cl, I,
Br, triflate, tosylate, nosylate, besylate or mesylate and all
other variables are as defined above.
[0308] For example, a bromide leaving group may be installed by
reacting a solution of the compound of formula (XII) in a solvent
such as dichloromethane, with carbon tetrabromide and
triphenylphosphine to prepare a compound of formula (XIII) wherein
X.sup.1 is Br. The compound of formula (XIII) may then be reacted
in solvent, such as tetrahydrofuran, with a compound of formula
(XIV) that has been pretreated with a base such as sodium hydride,
to prepare the compound of formula (V).
[0309] A compound of formula (V) may also be synthesized by the
condensation of a tricarbonyl compound of formula (XV) with
hydroxylamine according to the procedure of Doyle, F. P., et. al.,
1963 J. Chem. Soc. 5838-5845.
##STR00044## [0310] wherein all variables are as defined above.
[0311] A compound of formula (IV) wherein b is 2 or 3 may be
prepared by the homologation of a compound of the formula (IV)
wherein b is 1 (i.e., a compound of formula (IV-1), as depicted
below.
##STR00045## [0312] wherein all variables are as defined above.
[0313] More specifically, a compound of formula (IV) wherein b is 2
or 3 (formula IV-2 and IV-3, respectively) may be prepared by
oxidizing the compound of formula (IV-1) with an oxidizing agent
such as pyridium chlorochromate, to prepare the compound of formula
(XVI). The compound of formula (XVI) may be reacted with the ylide
formed from the reaction of (methoxymethyl)triphenyl-phosphonium
chloride and potassium tert butoxide to prepare a compound of
formula (XVII). The compound of formula (XVII) may be hydrolyzed
with aqueous hydrochloric acid to prepare the compound of formula
(XVIII). The compound of formula (XVIII) may be reduced with sodium
borohydride to prepare a compound of formula (IV-2) or may be
further homologated by the repeating the Wittig/hydrolysis sequence
to prepare a compound of formula (IV-3).
[0314] A compound of formula (II) may be prepared by coupling the
compound of formula (XXV) with a boronic acid or ester compound of
formula (XXVI) using conventional Suzuki coupling techniques.
##STR00046## [0315] wherein: X.sup.2 is chloro, bromo, iodo, or
triflate; [0316] R.sup.10 is H or alkyl; [0317] Z.sup.1 is --O-- or
--NH--; and [0318] all other variables are as defined above.
[0319] For example, the compound of formula (II) may be prepared by
coupling a compound of formula (XXV) with a compound of formula
(XXVI) in the presence of a suitable palladium complex such as
tetrakis(triphenylphosphine)palladium(0) and a base such as sodium
carbonate in a mixture of water and ethereal solvent such as
1,2-dimethoxyethane, at an elevated temperature.
[0320] A compound of formula (XXVI) may be synthesized by
techniques known to those skilled in the art or purchased
commercially.
[0321] A compound of formula (XXV) wherein Y.sup.1 is CH, Y.sup.2
is C--R.sup.1 where R.sup.1 is H or alkyl, Y.sup.3 is C--R.sup.1
where R.sup.1 is --CO.sub.2alkyl, and Y.sup.4 is N (i.e., formula
XXV-1) may be prepared by the steps of: [0322] a) reducing a
compound of formula (XXVII) with zinc (II) chloride and tin (II)
chloride to prepare a compound of formula (XXVIII), and [0323] b)
condensing the compound of formula (XXVIII) in situ with an alpha
ketoester of formula (XXIX). Suitable alpha ketoesters of formula
(XXIX) are commercially available or can be prepared using
conventional techniques known to those skilled in the art.
[0323] ##STR00047## [0324] wherein: R.sup.1a is H or alkyl, and all
other variables are as defined above.
[0325] A compound of formula (XVII) may be made by nitrating a
compound of formula (XXX) using nitric acid and concentrated
sulfuric acid.
##STR00048## [0326] wherein: all variables are as defined
above.
[0327] Suitable reaction conditions for such nitration are
conventional in the art. Compounds of formula (XXX) are
commercially available.
[0328] As another example, a compound of formula (XXV) wherein
Y.sup.1 and Y.sup.4 are both CH, Y.sup.2 is N and Y.sup.3 is
C--R.sup.1 where R.sup.1 is --CO.sub.2alkyl (i.e., formula XXV-2)
may be prepared by the steps of reacting a phenol of formula (XXXI)
with a reagent suitable for installing the leaving group,
X.sup.2.
##STR00049## [0329] wherein: X.sup.2 is triflate.
[0330] Reagents suitable for installing the leaving group include
but are not limited to triflic anhydride. The reaction may be
carried out in a solvent, such as dichloromethane and in the
presence of a suitable base, such as pyridine. In another
embodiment, a compound of formula (XXV-2) may be prepared by
reacting the compound of formula (XXXI) in a suspension of toluene
with an aqueous solution of tribasic potassium phosphate and then
reacting with triflic anhydride.
[0331] A compound of formula (XXXI) may be prepared by heating a
solution of a tetrahydroisoquinoline of formula (XXXII) in xylenes
with palladium on carbon.
##STR00050##
[0332] A tetrahydroquinoline of formula (XXXII) may be made
according to the procedure of K. Verschueren et al., 1992 Synthesis
458-460.
[0333] A quinoline compound of formula (XXV) wherein Y.sup.1 is N,
Y.sup.2 is CH, Y.sup.3 is C--R.sup.1 where R.sup.1 is
--CO.sub.2alkyl, Y.sup.4 is CH and R.sup.2 is H (i.e., formula
XXV-3) may be prepared by the steps of: [0334] a) reacting a
sulfonamide compound of formula (XXXIII) with
bis(acetyloxy)(phenyl)-.lamda..sup.3-iodane and iodine; and [0335]
b) reacting the resulting intermediate with a base, such as
potassium carbonate.
##STR00051##
[0336] Both reaction steps may be carried out at elevated
temperatures and in solvent such as dimethyl formamide.
[0337] An aminoester compound of formula (XXXIII) may be prepared
by the steps of: [0338] a) reacting a compound of formula (XXXIV)
with acetylchloride and a base, to prepare a diester compound of
formula (XXXV); and [0339] b) reacting the compound of formula
(XXXV) with 4-methylbenzenesulfonamide and a base.
[0339] ##STR00052## [0340] wherein Ac refers to acetyl.
[0341] Suitable bases for the step (a) of reacting the compound of
formula XXXIV with acetyl chloride or acetate include but are not
limited to triethylamine or pyridine.
[0342] The step of reacting the compound of formula XXXV with
methylbenzenesulfonamide may be carried out in a suitable solvent,
such as dimethylformamide with heating. Suitable bases for this
reaction include but are not limited to potassium carbonate, sodium
carbonate, or cesium carbonate.
[0343] A beta hydroxyester of formula of (XXXIV) may be prepared by
reacting an aromatic aldehyde of formula (XXXVI) with methyl
acrylate and a base, such as 1,4-diazobicyclo[2.2.2]octane
(DABCO).
##STR00053##
[0344] A quinoline compound of formula (XXV) wherein Y.sup.1 is N,
Y.sup.2 and Y.sup.3 are CH, Y.sup.4 is C--R.sup.1 where R.sup.1 is
--CO.sub.2alkyl and R.sup.2 is H (i.e., formula XXV-4) may be
prepared by the steps of: [0345] a) saponifying a diesaster of
formula (XXXVII) with a suitable base to prepare a diacid compound
of formula (XXXVIII); [0346] b) heating the compound of formula
(XXXVIII) and reesterifying the carboxylic acid to prepare a
compound of formula (XXV-4); [0347] c) reesterification of the
carboxylic acid with an alcohol and an acid such as sulfuric
acid.
##STR00054##
[0348] The saponification reaction may be carried out using
conventional reaction conditions. Suitable bases for the
saponification include but are not limited to sodium hydroxide. The
reaction may be carried out at ambient or elevated temperature. The
step of decarboxylating the compound of formula (XXXVIII) is
typically carried out in diphenyl ether. The re-esterifying
reaction is typically carried out in an appropriate alcoholic
solvent, such as methanol or ethanol, and an acid such as sulfuric
acid.
[0349] In another embodiment, a compound of formula (I) may be
prepared as depicted in Scheme 2.
##STR00055## [0350] wherein at least one R.sup.1 is
--CO.sub.2alkyl, and all other variables are as defined above.
[0351] In general, the process for preparing a compound of formula
(I) as depicted in Scheme 2 comprises the steps of: [0352] a)
reacting a compound of formula (IV) with a compound of formula (II)
under Mitsunobu reaction conditions to prepare a compound of
formula (I); [0353] b) optionally converting the compound of
formula (I) into a pharmaceutically acceptable salt or solvate
thereof; and [0354] c) optionally converting the compound of
formula (I) or a pharmaceutically acceptable salt or solvate
thereof into a different compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof.
[0355] More specifically, the compound of formula (I) is prepared
by reacting the compound of formula (IV) with a compound of formula
(II) under Mitsunobu reaction conditions. For example, a compound
of formula (I) can be prepared by the reacting a compound of
formula (II) with an alcohol of formula (IV) in a solution of
dichloromethane or toluene with triphenyl phosphine and a dialkyl
azodicarboxylate, such as diisopropyl azodicarboxylate or
di-tert-butyl azodicarboxylate. The compound of formula (IV) may be
prepared by processes described above.
[0356] In another embodiment, a compound of formula (I) wherein at
least one R.sup.1 is --CO.sub.2alkyl; d is 1; Z.sup.2 is --O--,
--S-- and --N(H)--; and Ring D is a moiety of formula i, ii or
iii:
##STR00056## [0357] (i.e., a compound of formula (I-A) may be
prepared as depicted in Scheme 3.
[0357] ##STR00057## [0358] wherein: at least one R.sup.1 is
--CO.sub.2alkyl; [0359] Z.sup.2a is selected from --O--, --S--,
--N(H)--, and --NC(O)CF.sub.3; [0360] Ring D is a moiety of formula
i, ii or iii:
##STR00058##
[0360] and all other variables are as defined above.
[0361] In general, the process of preparing a compound of formula
(I-A) according to Scheme 3 comprises the steps of: [0362] a)
reacting a compound of formula (XL) with acid to prepare a compound
of formula (XLI); [0363] b) reacting a compound of formula (XLI)
under Mitsunobu reaction conditions with a compound of formula i-a,
ii-a, or iii-a to prepare a compound of formula (I-A); [0364] c)
optionally converting the compound of formula (I-A) into a
pharmaceutically acceptable salt or solvate thereof; and [0365] d)
optionally converting the compound of formula (I-A) or a
pharmaceutically acceptable salt or solvate thereof into a
different compound of formula (I) or a pharmaceutically acceptable
salt or solvate thereof.
[0366] More specifically, a compound of formula (XL) may be
prepared by reacting the compound of formula (XL) with an acid. The
reaction may be carried out in a solvent, such as dichloromethane
or 1,2-dichloroethane. Suitable acids for use in this reaction will
be apparent to those skilled in the art and include, but are not
limited to trifluoroacetic acid. The resulting alcohol compound of
formula (XLI) may be reacted with a suitable Ring D moiety of
formula of i-a, ii-a, or iii-a under conventional Mitsunobu
reaction conditions. For example, this reaction may be carried out
in a solvent, such as dichloromethane or toluene, with triphenyl
phosphine and a dialkyl azodicarboxylate like diisopropyl
azodicarboxylate or di-tert-butyl azodicarboxylate to prepare a
compound of formula (I-A).
[0367] Upon hydrolysis of the ester to the acid the
trifluoroacetamide may be hydrolysed to the corresponding amine and
trifluoroacetic acid.
[0368] Compounds of formula (I) wherein d is 1 and Z.sup.2 is
--SO-- or --SO.sub.2-- may be prepared by oxidizing a compound of
formula (I-A) using a conventional oxidant, such as
3-chloroperbenzoic acid.
[0369] As another example, a compound of formula (I-A) may be
prepared as depicted in Scheme 3a.
##STR00059## [0370] wherein: [0371] X.sup.1 is chloride, iodide,
bromide, triflate, tosylate, nosylate, besylate or mesylate,
(preferably chloride); [0372] each R.sup.1 is the same or different
and is independently selected from alkyl, fluoroalkyl or
--CO.sub.2alkyl; wherein at least one R.sup.1 is --CO.sub.2alkyl;
[0373] c is 1; and [0374] all other variables are as defined
above.
[0375] In general, the process of preparing a compound of formula
(I-A) according to Scheme 3a comprises the steps of: [0376] a)
reacting a compound of formula (XLI) with a suitable reagent to
convert the alcohol to a leaving group, to prepare a compound of
formula (XLII); [0377] b) reacting a compound of formula (XLII)
with a suitable Ring D moiety of formula iv-a in the presence of
sodium hydride; [0378] c) optionally converting the compound of
formula (I-A) into a pharmaceutically acceptable salt or solvate
thereof; and [0379] d) optionally converting the compound of
formula (I-A) or a pharmaceutically acceptable salt or solvate
thereof into a different compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof.
[0380] More specifically, a compound of formula (XLII) may be
prepared by converting the alcohol compound of formula (XLI) into a
leaving group, X.sup.1 using conventional techniques. For example,
a solution of the compound of formula (XLI) in a solvent, such as
dichloromethane, may be reacted with a carbon tetrahalide, such as
carbon tetrabromide, followed by triphenylphosphine to provide a
compound of formula (XLII) wherein X.sup.1 is chloride, iodide, or
bromide. As another example, a solution of the compound of formula
(XLI) may be reacted with sulfonylchloride and a suitable base,
such as pyridine, to provide a compound of formula (XLII) wherein
X.sup.1 is triflate, tosylate, nosylate, besylate or mesylate.
Typically, the Ring D moiety is pretreated with a base such as
sodium hydride then reacted with a compound of formula (XLII) to
prepare the corresponding ether. The reaction of the compound of
formula (XLII) with the Ring D moiety may be carried out in a
solvent such as tetrahydrofuran.
[0381] In another embodiment, a compound of formula (I) may be
prepared as depicted in Scheme 4.
##STR00060## [0382] wherein: [0383] at least one R.sup.1 is
--CO.sub.2alkyl; [0384] R.sup.10 is H or alkyl; and [0385] all
other variables are as defined above.
[0386] In general, the process of Scheme 4 comprises the steps of:
[0387] a) reacting a compound of formula (XXV) with a boronic acid
or ester compound of formula (XLV) under Suzuki coupling conditions
to prepare a compound of formula (I); [0388] b) optionally
converting the compound of formula (I) into a pharmaceutically
acceptable salt or solvate thereof; and [0389] c) optionally
converting the compound of formula (I) or a pharmaceutically
acceptable salt or solvate thereof into a different compound of
formula (I) or a pharmaceutically acceptable salt or solvate
thereof.
[0390] More specifically, a compound of formula (I) may prepared
reacting a compound of formula (XXV) with a compound of formula
(XLV) under conventional Suzuki coupling reaction conditions. For
example, the reaction may be carried out in the presence of a
suitable palladium complex such as
tetrakis(triphenylphosphine)-palladium(0) and a base such as sodium
carbonate in a mixture of water and ethereal solvent such as
1,2-dimethoxyethane, at an elevated temperature. A compound of
formula (XXV) may be prepared as described above.
[0391] A compound of formula (XLV) may be prepared by reacting a
compound of formula (XXVI) with a compound of formula (III) in the
presence of a base such as cesium carbonate or potassium carbonate.
The reaction may be carried out in a polar aprotic solvent, such as
N,N-dimethylformamide.
##STR00061## [0392] wherein: [0393] X.sup.1 is chloro, iodo, bromo,
triflate, tosylate, nosylate, besylate or mesylate, (preferably
chloro); [0394] each R.sup.1 is the same or different and is
independently selected from alkyl, fluoroalkyl or --CO.sub.2alkyl;
wherein at least one R.sup.1 is 'CO.sub.2alkyl; and all other
variables are as defined above.
[0395] The boronic ester of formula (XLV) wherein R.sup.10 is
alkyl, may optionally be hydrolyzed to the corresponding boronic
acid if desired. A compound of formula (XXVI) may be synthesized by
techniques known to those skilled in the art or purchased
commercially. A compound of formula (III) may be prepared as
described above.
[0396] According to one method, a compound of formula (I) may be
prepared using the process depicted in Scheme 5, below.
##STR00062## [0397] wherein: [0398] at least one R.sup.1 is
C.sub.1-6alkyl or fluoroalkyl; [0399] R.sup.10 is H or alkyl;
[0400] Y.sup.2 is N; [0401] Y.sup.4 is N; and [0402] all other
variables are as defined above.
[0403] In general, the process of Scheme 5 comprises the steps of:
[0404] a) reacting a compound of formula (XXV-5) with a boronic
acid or ester compound of formula (XLV) under Suzuki coupling
conditions to prepare a compound of formula (I); [0405] b)
optionally converting the compound of formula (I) into a
pharmaceutically acceptable salt or solvate thereof; and [0406] c)
optionally converting the compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof into a
different compound of formula (I) or a pharmaceutically acceptable
salt or solvate thereof.
[0407] More specifically, a compound of formula (I) may be prepared
by reacting a compound of formula (XXV-5) with a compound of
formula (XLV) under conventional Suzuki coupling reaction
conditions. For example, the reaction may be carried out in the
presence of a suitable palladium complex such as
tetrakis(triphenylphosphine)-palladium(0) and a base such as sodium
carbonate in a mixture of water and ethereal solvent such as
1,2-dimethoxyethane, at an elevated temperature. A compound of
formula (XLV) may be prepared as described above.
[0408] A compound of formula (XXV-5) may be prepared by cyclizing a
compound of formula (LII) with a source of ammonia in acid, such as
ammonium acetate in acetic acid, at elevated temperatures.
##STR00063## [0409] wherein: [0410] R.sup.11 is selected from
C.sub.1-6alkyl or fluoroalkyl; [0411] all other variables are as
defined above.
[0412] A compound of formula (LII) may be prepared by reacting an
alkyl chlorooxoacetate of formula (L), like ethyl chlorooxoacetate,
with an aniline of formula (LI) in the presence of a base like
pyridine. Alkyl chlorooxoacetates of formula (L) are commercially
available. Anilines of formula (LI) can be made according to
techniques well known in the art.
##STR00064## [0413] wherein: [0414] R.sup.11 is a selected from H,
alkyl, fluoroalkyl, --CO.sub.2H and --CO.sub.2alkyl; [0415] all
other variables are as defined above.
[0416] According to one method, a compound of formula (I) may be
prepared using the process depicted in Scheme 6, below.
##STR00065## [0417] wherein: [0418] R.sup.10 is H or alkyl; [0419]
Y.sup.1 is N; [0420] Y.sup.2 is CH; [0421] Y.sup.4 is N; and [0422]
all other variables are as defined above.
[0423] In general, the process of Scheme 6 comprises the steps of:
[0424] a) reacting a compound of formula (XXV-6) with a boronic
acid or ester compound of formula (XLV) under Suzuki coupling
conditions to prepare a compound of formula (I); [0425] b)
optionally converting the compound of formula (I) into a
pharmaceutically acceptable salt or solvate thereof; and [0426] c)
optionally converting the compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof into a
different compound of formula (I) or a pharmaceutically acceptable
salt or solvate thereof.
[0427] More specifically, a compound of formula (I) may be prepared
by reacting a compound of formula (XXV-6) with a compound of
formula (XLV) under conventional Suzuki coupling reaction
conditions. For example, the reaction may be carried out in the
presence of a suitable palladium complex such as
tetrakis(triphenylphosphine)-palladium(0) and a base such as sodium
carbonate in a mixture of water and ethereal solvent such as
1,2-dimethoxyethane, at an elevated temperature.
[0428] A compound of formula (XLV) may be prepared as described
above.
[0429] A compound of formula (XXV-6) may be prepared by reacting
4-bromo-o-phenylenediamine with an alkylbromopyruvate in a solvent
like 1-methyl-2-pyrrolidinone.
##STR00066##
[0430] As will be apparent to those skilled in the art, a compound
of formula (I) may be converted to a different compound of formula
(I) using techniques already described above and other techniques
well known in the art. For example, compound of formula (I) wherein
R.sup.1 is --CO.sub.2alkyl made be converted to the corresponding
acid (i.e. a compound of formula (I) wherein R.sup.1 is
--CO.sub.2H) by saponification with a suitable base such as sodium
hydroxide in a solution of water and tetrahydrofuran and optionally
an alcoholic co-solvent.
[0431] A compound of formula (I) wherein R.sup.1 is CO.sub.2H (I-B)
may be reacted with an amine to prepare the corresponding amide
(i.e. a compound of formula (I) wherein R.sup.1 is --C(O)NH.sub.2,
hereinafter a compound of formula (I-C)). This reaction may be
carried out using conventional techniques. For example, a compound
of formula (I-B) may be reacted with di-tert-butyl dicarbonate in
acetonitrile with a base such as pyridine then ammonium hydrogen
carbonate to produce a compound of formula (I-C). See, C. D.
Haffner, et al., US2004/0171848.
[0432] A compound of formula (I-C) may be dehydrated to prepare a
corresponding nitrile. This reaction may be carried out using
conventional amide dehydration techniques. For example, a compound
of formula (I-C) may be dehydrated with phosphorous oxychloride in
a solution of methylene chloride and a suitable base such as
triethylamine to prepare the corresponding nitrile analogue of the
compound of formula (I). (Uiterweerd, P. G. H., et al., 2003
Tetrahedron: Asymmetry, 14:3479-3485). The nitrile analogue of the
compound of formula (I) may be condensed with sodium azide to
prepare the corresponding tetrazole (i.e. a compound of formula (I)
wherein R.sup.1 is the acid-equivalent group tetrazole; hereinafter
a compound of formula (I-D)). This reaction may be carried out
using conventional techniques. For example, a compound of formula
(I-D) may be prepared by reacting the nitrile analogue of the
compound of formula (I) with sodium azide in the presence of
ammonium chloride in a polar aprotic solvent such as
N,N-dimethylformamide at an elevated temperature. (Meyer, E., et
al., 2003 Synthesis 899-905). Other techniques for converting the
nitrile analogue of the compound of formula (I) into compounds of
formula (I) wherein R.sup.1 is the acid-equivalent group are known
in the art. See, Ellingboe, J. W., et al., 1993 J. Med. Chem.
36:2485-2493 and Weller, H. N., et al., 1993 Heterocycles
36:1027-1038.
[0433] Based upon these examples and the disclosure contained
herein one skilled in the art can readily convert compounds of
formula (I) into other compounds of formula (I), or salts or
solvates thereof.
[0434] The following examples are intended for illustration only
and are not intended to limit the scope of the invention in any
way, the present invention being defined by the claims.
[0435] In the examples, the following terms have the designated
meaning: [0436] .ANG.=angstrom [0437] g=gram; [0438] mg=milligram;
[0439] mol=mole; [0440] mmol=millimole; [0441] M=molar; [0442]
N=normal; [0443] L=liter; [0444] mL=milliliter; [0445]
.mu.L=microliter; [0446] hr=hour; [0447] min=minute; [0448]
aq=aqueous; [0449] wt %=(weight percent) [0450]
CH.sub.2Cl.sub.2=dichloromethane; [0451] DMF=N,N-dimethylformamide;
[0452] DMSO=dimethylsulfoxide; [0453] DTT=dithiothreitol [0454]
EtOAc=ethyl acetate; [0455] EtOH=ethanol [0456] HCl=hydrogen
chloride; [0457] H.sub.2O=water; [0458] K.sub.2CO.sub.3=potassium
carbonate; [0459] K.sub.3PO.sub.4=potassium phosphate [0460]
MeOH=methanol [0461] MgSO.sub.4=magnesium sulfate; [0462]
N.sub.2=nitrogen; [0463] Na.sub.2CO.sub.3=sodium carbonate; [0464]
NaF=sodium fluoride [0465] NaHCO.sub.3=sodium hydrogen carbonate;
[0466] NaOH=sodium hydroxide; [0467] Na.sub.2SO.sub.4=sodium
sulfate; [0468] P.sub.2O.sub.5=phosphorus pentoxide; [0469]
SiO.sub.2=silicon dioxide; [0470] TEA=triethylamine; [0471]
THF=tetrahydrofuran; [0472] NMR=nuclear magnetic resonance; [0473]
H=Hydrogen; [0474] Hz=hertz; MHz=megahertz [0475] OD=optical
density; [0476] HPLC=high performance liquid chromatography;
[0477] APCI-LCMS=Atmospheric Pressure Chemical Ionization-Liquid
Chromatography Mass Spectrometry; ESI-LCMS=Electrospray
Ionization-Liquid Chromatography Mass Spectrometry; HRMS=High
Resolution Mass Spectrometry.
Example 1
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-2-quinolinecarboxylic Acid
##STR00067##
[0478] 1a) 6-Bromo-2-(tribromomethyl)quinoline
##STR00068##
[0480] A suspension of 6-bromo-2-methylquinoline (175 g, 0.79 mol)
and sodium acetate (350 g, 4.3 mol) in acetic acid (1.5 L) was
stirred and heated in a 75.degree. C. bath until a solution formed.
A solution of bromine (132 mL, 2.6 mol) in acetic acid (350 mL) was
added over 15 min during which time the reaction temperature rose
to 86.degree. C. The resulting suspension was heated at 120.degree.
C. for 1 hr. The suspension was cooled to 80.degree. C. and added
to ice-water (6 L) with stirring. The resulting white solid was
collected by filtration, washed with water (2.times.1 L) and air
dried to give 6-bromo-2-(tribromomethyl)quinoline (350 g, 95%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.54 (d, J=9 Hz, 1H),
8.40 (d, J=2 Hz, 1H), 8.31 (d, J=9 Hz, 1H), 8.00 (m, 2H). ES-LCMS
m/z 459 (M+H).sup.-.
1b) 6-Bromo-2-quinolinecarboxylic Acid
##STR00069##
[0482] Concentrated sulfuric acid (0.75 L) was added during 15 min
to a stirred suspension of 6-bromo-2-(tribromomethyl)quinoline (350
g, 0.76 mol) in water (1.75 L). The resulting suspension was heated
at 150.degree. C. (bath temperature) for 5 hr. The mixture was
cooled and the precipitate was collected by filtration, washed with
water and dried to give 6-bromo-2-quinolinecarboxylic acid as a
solid (127.6 g). The filtrate was diluted with water (3 L) and a
second crop of the product was obtained (55.7 g, combined yield
183.3 g, 96%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.50
(d, J=9 Hz, 1H), 8.38 (d, J=2 Hz, 1H), 8.13 (d, J=9 Hz, 1H), 8.06
(d, J=9 Hz, 1H), 7.96 (m, 1H). ES-LCMS m/z 253 (M+H).sup.+.
1c) Methyl 6-bromo-2-quinolinecarboxylate
##STR00070##
[0484] A mixture of 6-bromo-2-quinolinecarboxylic acid (331 g, from
multiple batches, 1.31 mol) and methanesulfonic acid (22 mL, 33 g,
0.34 mol) in methanol (2 L) was refluxed for 6 hr. The mixture was
treated with a solution of sodium bicarbonate (29 g, 0.34 mol) in
water (350 mL) and the resulting suspension was slowly cooled to
20.degree. C. and stirred overnight. The suspension was filtered
and the cake was washed with water (1 L). The solid was dried in a
vacuum oven at 50.degree. C. for 3 days to yield methyl
6-bromo-2-quinolinecarboxylate (294 g, 85%). .sup.1H NMR (400 MHz,
DMSO-d.sub.6); .delta. 8.52 (d, J=9 Hz, 1H), 8.40 (d, J=2 Hz, 1H),
8.14 (d, J=9 Hz, 1H), 8.08 (d, J=9 Hz, 1H), 7.97 (m, 1H), 3.93 (s,
3H). ES-LCMS m/z 267 (M+H).sup.+.
1d) Methyl 6-(4-Hydroxyphenyl)-2-quinolinecarboxylate
##STR00071##
[0486] A stirred mixture of methyl 6-bromo-2-quinolinecarboxylate
(297 g, from multiple batches, 1.11 mol), 4-hydroxybenzeneboronic
acid (184 g, 1.33 mol), 2 M K.sub.3PO.sub.4 (1.8 L) and
tetrakistriphenylphosphine palladium (0) (51.3 g, 0.04 mol) in
1,2-dimethoxyethane (1.8 L) was heated to 79.degree. C. A solution
formed and the reaction was complete. The mixture was cooled to
45.degree. C., diluted with water (1.8 L), cooled to 15.degree. C.
and held at this temperature for 30 minutes then filtered. The
filter cake was washed with methanol (4.times.2 L) to yield methyl
6-(4-hydroxyenyl)-2-quinolinecarboxylate (83 g). HPLC analysis
showed that much of the ester was hydrolyzed. The mixture was
acidified to pH 3 by adding 6 N HCl and the resulting precipitate
was collected by filtering, washed with water and dried at
45.degree. C. in a vacuum oven to yield mostly
6-(4-hydroxyphenyl)-2-quinolinecarboxylic acid (116.3 g). The
methanol wash was concentrated, azetroped with toluene and dried
under vacuum to yield the acid phenol (178.5 g). This material was
re-esterified by refluxing with methanesulfonic acid (71 mL, 105.7
g, 1.1 mol) in methanol (1.8 L) for 22 hours. The mixture was
cooled to room temperature, diluted with water (5 L) and basified
to pH 8 by adding NaHCO.sub.3 solution. The resulting suspension
was filtered and the filter cake was dried at 50.degree. C. in a
vacuum oven to yield methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (222.3 g, combined yield
305 g, 98%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 9.75 (br
s, 1H), 8.53 (d, J=9 Hz, 1H), 8.25 (s, 1H), 8.11 (m, 3H), 7.69 (d,
J=9 Hz, 2H), 6.90 (d, J=9 Hz, 2H), 3.93 (s,3H). ES-LCMS m/z 280
(M+H).sup.+.
1e)
4-(Chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
##STR00072##
[0488] Thionyl chloride (123 mL, 202 g, 1.7 mol) was added dropwise
during 30 min to a stirred suspension of benzotriazole (202 g, 1.7
mol) in dichloromethane (550 mL) at room temperature under N.sub.2.
The resulting yellow solution was transferred to an addition funnel
and added dropwise during 1 hour to a stirred solution of
[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methanol
(372 g, 1.3 mol, Maloney, P. R., et al., 2000 J. Med. Chem.
43:2971-2974) in dichloromethane (975 mL). The reaction temperature
gradually rose to a maximum of 28.degree. C. After 1 hr the
resulting suspension was filtered to remove the benzotriazole
hydrochloride. The filtrate was washed with water (2.times.1 L),
with 1 N NaOH (1 L), with water (1 L), dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated to yield
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
as a pale yellow oil (413 g, 80%). .sup.1H NMR (400 MHz,
DMSO-d.sub.6); .delta. 7.64 (m, 3H), 4.47 (s, 2H), 3.45 (m, 1H),
1.31 (d, J=7 Hz, 6H). ES-LCMS m/z 305 (M+H).sup.+.
1f) Methyl
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-2-quinolinecarboxylate
##STR00073##
[0490] A mixture of methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (222 g, 0.8 mol),
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(292 g, 0.96 mol) and cesium carbonate (312 g, 0.96 mol) in DMF
(1.5 L) was heated at 65.degree. C. for 28 hours. Additional
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(105 g, 0.34 mol) dissolved in DMF (150 mL) was added. The mixture
was cooled to room temperature and poured into water (5 L) with
stirring. The resulting taffy was separated from the supernatant
and stirred with ethyl acetate (2.5 L) for 3 hours at room
temperature. The resulting suspension was filtered and the filter
cake was dried to yield methyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylate as a solid (245 g, 56%). .sup.1H NMR
(400 MHz, DMSO-d.sub.6): .delta. 8.54 (d, J=9 Hz, 1H), 8.28 (d, J=1
Hz, 1H), 8.11 (m, 3H), 7.72 (d, J=9 Hz, 2H), 7.62 (d, J=8 Hz, 2H),
7.53 (m, 1H), 6.93 (d, J=9 Hz, 2H), 4.88 (s, 2H), 3.93 (s, 3H),
3.47 (m, 1H), 1.33 (d, J=7 Hz, 6H). ES-LCMS m/z 547
(M+H).sup.+.
1g)
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}o-
xy)phenyl]-2-quinolinecarboxylic acid
##STR00074##
[0492] A stirred mixture of methyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylate (245 g, 0.45 mol), NaOH (36 g, 0.90
mol) dissolved in water (225 mL), methanol (0.5 L) and THF (1.2 L)
was heated at 65.degree. C. for 90 min. The mixture was cooled to
room temperature, treated with 6 N HCl (150 mL) and stirred at room
temperature for 30 min. The resulting precipitate was collected by
filtering, washed with water (2.times.1 L) and dried overnight at
45.degree. C. in a vacuum oven to yield
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid (162 g, 68%). A second
crop of material was similarly obtained (27.2 g, 11%). .sup.1H NMR
(400 MHz, DMSO-d.sub.6): .delta. 8.48 (d, J=9 Hz, 1H), 8.24 (d, J=2
Hz, 1H), 8.17 (d, J=9 Hz, 1H), 8.09 (d, J=9 Hz, 2H), 7.72 (d, J=9
Hz, 2H), 7.62(d, J=8 Hz, 2H), 7.53 (m, 1H), 6.92 (d, J=9 Hz, 2H),
4.88 (s, 2H), 3.46 (m, 1H), 1.32 (d, J=7 Hz, 6H). ES-LCMS m/z 533
(M+H).sup.-.
1h) Potassium
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic Acid
##STR00075##
[0494] A stirred suspension of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid (179 g, 0.335 mol) in ethanol
(2.6 L) was treated with 1 N potassium tert-butoxide in tent-butyl
alcohol (350 mL) and stirred at room temperature under N.sub.2 for
1 hour. The mixture was then heated to a gentle reflux for 2 hours
and was stirred overnight at room temperature. The suspension was
filtered, then the filter cake was washed with hexane, combined
with 5 g of similarly prepared material and dried at 60.degree. C.
in a vacuum oven overnight to yield
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid potassium salt as a beige solid
(181 g, 84%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.25 (d,
J=9 Hz, 1H), 8.18 (d, J=9 Hz, 1H), 8.08 (d, J=2 Hz, 1H), 8.02 (d,
J=8 Hz, 1H), 7.94 (dd, J=2, 9 Hz, 1H), 7.68 (d, J=9 Hz, 2H), 7.62
(d, J=8 Hz, 2H), 7.53 (dd, J=7, 9 Hz, 1H), 6.90 (d, J=9 Hz, 2H),
4.86 (s, 2H), 3.46 (m, 1H), 1.32 (d, J=7 Hz, 6H). ES-LCMS m/z 533
(M+H).sup.+.
[0495] Anal. calcd. for
C.sub.29H.sub.21Cl.sub.2KN.sub.2O.sub.4.0.25 KCl: C, 59.02; H,
3.59; Cl, 13.52; N, 4.75. Found: C, 58.94; H, 3.62; Cl, 13.28; N,
4.74.
1i) Sodium
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-2-quinolinecarboxylate
##STR00076##
[0497] To a stirring suspension of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid (4.12 g, 7.73 mmol) in methanol
(100 mL) was added 1 N sodium hydroxide (7.73 mL, 7.73 mmol). The
mixture was stirred for approximately 1 hour and then concentrated.
Ether was added followed by a little water. After stirring the
solid turned into a paste. The solvent was decanted and the paste
triturated with ether and then dried on a rotary evaporator at
35.degree. C. under vacuum for approximately 1.5 hours to afford
sodium
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylate (4.24 g, 98%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.28-8.22 (m, 2H), 8.09-7.96 (m, 3H), 7.68
(d, J=9 Hz, 2H), 7.63-7.51 (m, 3H), 6.90 (d, J=9 Hz, 2H), 4.87 (s,
2H), 3.46 (septet, J=7 Hz, 1H), 1.33 (d, J=7 Hz, 6H). LRMS (APCI)
C.sub.29H.sub.21Cl.sub.2N.sub.2O.sub.4Na calculated: 533
(M+H).sup.+, found: 533 (M+H).sup.+.
Example 2
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-3-quinolinecarboxylic acid
##STR00077##
[0498] 2a) Methyl
2-[(4-bromophenyl)(hydroxy)methyl]-2-propenoate
##STR00078##
[0500] A solution of 2.46 g (13.3 mmol) of 4-bromobenzaldehyde, 1.0
mL (11.1 mmol) of methyl acrylate and 125 mg (1.11 mmol) of
1,4-diazobicyclo[2.2.2]octane in 5 mL Me0H was stirred at ambient
temperature for 46 hr. The solvent was evaporated and the residue
taken up in EtOAc. The organics were washed with 0.5 N HCl (aq),
then with brine, and concentrated. The residue was purified by
silica gel chromatography (120 g of silica gel eluting with 0-30%
EtOAc in hexanes over 45 minutes) to give 1.42 g (40%) of methyl
2-[(4-bromophenyl)(hydroxy)methyl]-2-propenoate as a clear oil.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.47 (d, J=2 Hz, 2H),
7.27 (d, J=2 Hz, 2H), 6.33 (s, 1H), 5.82 (s, 1H), 5.51 (d, J=5 Hz,
1H), 3.72 (s, 3H), 3.07 (d, J=5 Hz, 1H). ESI-LCMS m/z 293
(M+Na).sup.+.
2b) Methyl
(2E)-3-(4-bromophenyl)-2-({[(4-methylphenyl)sulfonyl]amino}meth-
yl)-2-propenoate
##STR00079##
[0502] To a solution of 390 .mu.L (5.46 mmol) of acetyl chloride in
10 mL of THF at 0.degree. C. was added 1.41 g (5.20 mmol) of methyl
2-[(4-bromophenyl)(hydroxy)methyl]-2-propenoate and 870 .mu.L (6.24
mmol) of TEA in 10 mL THF. After 2 hr at ambient temperature
another 870 .mu.L (6.24 mmol) of TEA and 390 .mu.L (5.46 mmol) of
acetyl chloride was added and the solution stirred at ambient
temperature for an additional 1 hr. EtOAc was added and the
organics washed with three 50 mL portions of H.sub.2O then 50 mL of
brine. The organics were dried over Na.sub.2SO.sub.4 then
concentrated. This residue was then taken up in 25 mL of DMF and
4.35 g (25.4 mmol) of 4-methylbenzenesulfonamide and 3.51 g (25.4
mmol) of K.sub.2CO.sub.3 were added. The mixture was stirred at
50.degree. C. for 3 hr. To the mixture was added 100 mL of EtOAc
which was then washed with three 50 mL portions of water, then
brine. The solution was concentrated and the residue purified by
silica gel chromatography (120 g of silica gel eluting with 0-40%
EtOAc in hexanes over 45 minutes) to give 820 mg (37%) of methyl
(2E)-3-(4-bromophenyl)-2-({[(4-methylphenyl)sulfonyl]amino}methyl)-2-prop-
enoate as a beige glass. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
7.68-7.66 (m, 3H), 7.52 (d, J=7 Hz, 2H), 7.29-7.25 (m, 4H), 5.51
(t, J=6 Hz, 1H), 3.86 (d, J=6 Hz, 2H), 3.75 (s, 3H), 2.43 (s, 3H).
ESI-LCMS m/z 425 (M+H).sup.+.
2c) Methyl 7-bromo-3-quinolinecarboxylate
##STR00080##
[0504] A solution of 820 mg (1.93 mmol) of methyl
(2E)-3-(4-bromophenyl)-2-({[(4-methylphenyl)sulfonyl]amino}methyl)-2-prop-
enoate, 995 mg (3.09 mmol) of
bis(acetyloxy)(phenyl)-.lamda..sup.3-iodane and 490 mg (1.93 mmol)
of iodine in 35 mL of 1,2-dichloroethane were stirred at 70.degree.
C. for 30 min. The solvent was evaporated and the residue taken up
in 25 mL DMF and 1.07 g (7.73 mmol) of K.sub.2CO.sub.3 was added.
The mixture was stirred at 120.degree. C. for 6 hr. EtOAc was added
and the organics were washed with three portions of H.sub.2O then
brine. The solution was then concentrated and the residue purified
by silica gel chromatography (40 g of silica gel eluting with 0-40%
EtOAc in hexanes over 45 minutes) to give 80 mg (16%) of methyl
7-bromo-3-quinolinecarboxylate as an off-white solid. .sup.1H NMR
(400 MHz, CDCl.sub.3): .delta. 9.34 (s, 1H), 8.81 (s, 1H), 8.35 (s,
1H), 7.80 (d, J=9 Hz, 1H), 7.73 (d, J=9 Hz, 1H), 4.03 (s, 3H).
ESI-LCMS m/z 267 (M+H).sup.+.
2d) Methyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-3-quinolinecarboxylate
##STR00081##
[0506] A solution of 78 mg (0.29 mmol) of methyl
7-bromo-3-quinolinecarboxylate, 97 mg (0.44 mmol) of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol, 3 mg (0.01
mmol) of palladium acetate, 8 mg (0.03 mmol) of triphenylphosphine,
218 mg (1.03 mmol) of K.sub.3PO.sub.4 and 25 .mu.L of H.sub.2O in
1.0 mL of dioxane was stirred at 60.degree. C. for 1 hr. EtOAc was
added and the organics were washed with water and brine then
concentrated. To the residue was added 65 mg (0.21 mmol) of
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole,
74 mg (0.54 mmol) of K.sub.2CO.sub.3 and 1.5 mL of DMF and the
mixture was then stirred at 60.degree. C. for 1 hr. EtOAc was added
and the organics were washed with three portions of water, then
brine. The solution was concentrated and the residue purified by
silica gel chromatography (12 g of silica gel eluting with 0-40%
EtOAc in hexanes over 45 minutes) to give 47 mg (30%) of methyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-quinolinecarboxylate as a clear glass. .sup.1H NMR (400
MHz, CDCl.sub.3): .delta. 9.44 (s, 1H), 8.83 (s, 1H), 8.28 (s, 1H),
7.96 (d, J=9 Hz, 1H), 7.82 (d, J=9 Hz, 1H), 7.64 (d, J=9 Hz, 2H),
7.41 (d, J=8 Hz, 2H), 7.35-7.31 (m, 1H), 6.92 (d, J=9 Hz, 2H), 4.79
(s, 2H), 4.02 (s, 3H), 3.38-3.34 (m, 1H), 1.45 (d, J=7 Hz, 6H).
ESI-LCMS m/z 548 (M+H).sup.+.
2e)
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}o-
xy)phenyl]-3-quinolinecarboxylic acid
##STR00082##
[0508] To 47 mg (0.09 mmol) of methyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-quinolinecarboxylate in a mixture of 3 mL of EtOH, 1 mL
THF and 1 mL H.sub.2O was added 35 mg (0.86 mmol) of NaOH and the
mixture stirred at 50.degree. C. for 16 hr. The mixture was
concentrated to 1/3 volume then added dropwise to a a stirred
solution of 10 mL of 0.5 M HC1(aq.). The resulting solids were
collected by suction filtration, washed with water then dried to
yield 32 mg (70%) of
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-quinolinecarboxylic acid as a yellow solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6): 6 9.30 (s, 1H), 8.97 (s, 1H), 8.24-8.21
(m, 2H), 7.99 (d, J=8 Hz, 1H), 7.76 (d, J=7.61 (m, 2H), 7.55-7.52
(m, 1H), 6.93 (d, J=9 Hz, 2H), 4.88 (s, 2H), 3.50-3.44 (septet, J=7
Hz, 1H), 1.33 (d, J=7 Hz, 6H). ESI-LCMS m/z 534 (M+H).sup.+.
Example 3
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-2,4-quinolinedicarboxylic acid
##STR00083##
[0509] 3a) Dimethyl
7-(4-hydroxyphenyl)-2,4-quinolinedicarboxylate
##STR00084##
[0511] A mixture of 150 mg (0.46 mmol) of dimethyl
7-bromo-2,4-quinolinedicarboxylate synthesized according the
procedure of Corey, E. J. and Tramontano, A., 1981 J. Am. Chem.
Soc. 103:5599-5600 and Carrigan, C. N., et al, 1999 Bioorg. Med.
Chem. Lett. 9:2607-2612, 153 mg (0.69 mmol) of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol, 5 mg (0.02
mmol) of palladium acetate, 12 mg (0.05 mmol) of
triphenylphosphine, 345 mg (1.62 mmol) of K.sub.3PO.sub.4, and 40
.mu.L (2.31 mmol) of H.sub.2O in 3 mL of dioxane were stirred at
60.degree. C. for 1.5 hr. EtOAc was then added and the solution
washed with H.sub.2O. The aqueous layer was extracted with EtOAc
and then the combined organics were washed with brine and
concentrated. A small amount of cold EtOAc was added to the residue
and the solids were collected by suction filtration, washed with
cold EtOAc then dried to yield 79 mg (51%) of dimethyl
7-(4-hydroxyphenyl)-2,4-quinolinedicarboxylate as a brown solid.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.79 (s, 1H), 8.72 (d,
J=9 Hz, 1H), 8.41-8.38 (m, 2H), 8.17 (d, J=9 Hz, 1H), 7.77 (d, J=8
Hz, 2H), 6.91 (d, J=8 Hz, 2H), 4.00 (s, 3H), 3.97 (s, 3H). ESI-LCMS
m/z 338 (M+H).sup.+.
3b) Dimethyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2,4-quinolinedicarboxylate
##STR00085##
[0513] A solution of 75 mg (0.22 mmol) of dimethyl
7-(4-hydroxyphenyl)-2,4-quinolinedicarboxylate, 135 mg (0.44 mmol)
of
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
and 125 mg (0.89 mmol) of K.sub.2CO.sub.3 in 2 mL DMF was stirred
at ambient temperature for 60 hr. EtOAc was added and the mixture
washed with three portions of H.sub.2O then brine. The solution was
concentrated and the residue purified by silica gel chromatography
(12 g of silica gel eluting with 0-50% EtOAc in hexanes over 45
minutes) to give 62 mg (46%) of dimethyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]me-
thyl}oxy)phenyl]-2,4-quinolinedicarboxylate as a clear glass.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.88 (d, J=9.0 Hz, 1H),
8.65 (s, 1H), 8.50 (s, 1H), 7.98 (d, J=9 Hz, 1H), 7.65 (d, J=9 Hz,
2H), 7.41 (d, J=8 Hz, 2H), 7.35-7.31 (m, 1H), 6.91 (d, J=9 Hz, 2H),
4.79 (s, 2H), 4.11 (s, 3H), 4.07 (s, 3H), 3.36 (septet, J=7 Hz,
1H), 1.44 (d, J=7 Hz, 6H). ESI-LCMS m/z 606 (M+H).sup.+.
3c)
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}o-
xy)phenyl]-2,4-quinolinedicarboxylic acid
##STR00086##
[0515] To a solution of 60 mg (0.10 mmol) of dimethyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2,4-quinolinedicarboxylate in a mixture of 1 mL THF, 3 mL
EtOH, and 1 mL H.sub.2O was added 40 mg (0.99 mmol) of NaOH. The
mixture was stirred at 60.degree. C. for 16 hr. H.sub.2O (1.5 mL)
was added to dissolve some solids then the solution was
concentrated to 1/3 volume and added dropwise to 10 mL of 0.5 N HCl
(aq). After sitting for 5 min, the resulting solids were collected
by suction filtration, washed with H.sub.2O and dried to yield 36
mg (63%) of
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2,4-quinolinedicarboxylic acid as an orange solid. .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. 8.82 (d, J=9 Hz, 1H), 8.41 (s,
1H), 8.38 (s, 1H), 8.14 (dd, J.sub.A=2 Hz, J.sub.B=9 Hz, 1H), 7.79
(d, J=9 Hz, 2H), 7.61 (d, J=8 Hz, 2H), 7.55-7.51 (m, 1H), 6.94 (d,
J=9 Hz, 2H), 4.88 (s, 2H), 3.45 (septet, J=7 Hz, 1H), 1.34 (d, J=7,
6H). ESI-LCMS m/z 578 (M+H).sup.+.
Example 4
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-4-quinolinecarboxylic acid
##STR00087##
[0516] 4a) Methyl 7-bromo-4-quinolinecarboxylate
##STR00088##
[0518] To a solution of 750 mg (2.31 mmol) of dimethyl
7-bromo-2,4-quinolinedicarboxylate in a mixture of 20 mL of EtOH,
10 mL THF and 4 mL of H.sub.2O was added 925 mg (23.1 mmol) of
NaOH. The mixture was stirred at 50.degree. C. An additional 10 mL
of THF and 15 mL of H.sub.2O were added after 5 min. After 30 min
total at 50.degree. C., the solution was concentrated to 1/3 volume
and the pH was adjusted to 4.0 with 1.0 N HCl (aq) followed by the
addition of 100 mL of H.sub.2O. The resulting solids were collected
by suction filtration, washed with H.sub.2O and dried. The solids
were then added to 10 mL of diphenyl ether and the mixture stirred
at 215.degree. C. for 20 min. Upon cooling, 20 mL of hexanes was
added and the resulting solids were collected by suction
filtration, washed with hexanes and dried. The solids were then
suspended in 20 mL of MeOH and 505 .mu.L (6.90 mmol) of thionyl
chloride was added. The mixture was refluxed for 1 hr, then an
additional 505 .mu.L (6.90 mmol) of thionyl chloride was added.
After an additional 1 hr of refluxing, the solvent was evaporated.
The residue was diluted with EtOAc then washed with saturated
Na.sub.2CO.sub.3 (aq) and brine then dried over Na.sub.2SO.sub.4.
Concentration of the solution yielded 320 mg (52%) of methyl
7-bromo-4-quinolinecarboxylate as a beige solid. .sup.1H NMR (400
MHz, CDCl.sub.3): .delta. 9.01 (d, J=4 Hz, 1H), 8.70 (d, J=9 Hz,
1H), 8.35 (s, 1H), 7.92 (d, J=4 Hz, 1H), 7.75-7.72 (m, 1H), 4.03
(s, 3H). ESI-LCMS m/z 267 (M+H).sup.+.
4b) Methyl 7-(4-hydroxyphenyl)-4-quinolinecarboxylate
##STR00089##
[0520] A mixture of 160 mg (0.60 mmol) of methyl
7-bromo-4-quinolinecarboxylate, 200 mg (0.90 mmol) of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol, 7 mg (0.03
mmol) of palladium(II) acetate, 16 mg (0.06 mmol) of
triphenylphosphine, 450 mg (2.10 mmol) of K.sub.3PO.sub.4 and 50 uL
(3.01 mmol) of H.sub.2O in 3 mL of dioxane was stirred at
60.degree. C. for 1.5 hr. EtOAc was added then the organics were
washed with H.sub.2O and brine then concentrated. The residue was
suspended in cold EtOAc and the solids were collected by suction
filtration, washed with cold EtOAc and dried to give 92 mg (55%) of
methyl 7-(4-hydroxyphenyl)-4-quinolinecarboxylate as a tan solid.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.75 (s, 1H), 9.03 (s,
1H), 8.63 (d, J=9 Hz, 1H), 8.25 (d, J=2 Hz, 1H), 8.03 (dd,
J.sub.A=2 Hz, J.sub.B=9 Hz, 1H), 7.87 (s, 1H), 7.72 (d, J=8 Hz,
2H), 6.90 (d, J=8 Hz, 2H), 3.98 (s, 3H). ESI-LCMS m/z 280
(M+H).sup.+.
4c) Methyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-4-quinolinecarboxylate
##STR00090##
[0522] A solution of 90 mg (0.32 mmol) of methyl
7-(4-hydroxyphenyl)-4-quinolinecarboxylate, 196 mg (0.64 mmol) of
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
and 178 mg (1.29 mmol) of K.sub.2CO.sub.3 in 2.5 mL DMF was stirred
at ambient temperature for 60 hr. EtOAc was added and the mixture
washed with three portions of H.sub.2O then brine. The solution was
concentrated and the residue purified by silica gel chromatography
(12 g of silica gel eluting with 0-50% EtOAc in hexanes over 45
minutes) to give 98 mg (56%) of methyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-4-quinolinecarboxylate as a clear glass.
[0523] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.02 (d, J=4 Hz,
1H), 8.81 (d, J=9 Hz, 1H), 8.29 (s, 1H), 7.88-7.85 (m, 2H), 7.64
(d, J=8 Hz, 2H), 7.41 (d, J=8 Hz, 2H), 7.35-7.32 (m, 1H), 6.91 (d,
J=8 Hz, 2H), 4.79 (s, 2H), 4.05 (s, 3H), 3.34 (septet, J=7 Hz, 1H),
1.43 (d, J=7 Hz, 6H). ESI-LCMS m/z 548 (M+H).sup.-.
4d)
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}o-
xy)phenyl]-4-quinolinecarboxylic acid
##STR00091##
[0525] To 96 mg (0.48 mmol) of methyl
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-quinolinecarboxylate in a mixture of 1 mL of THF, 3 mL of
EtOH, and 1 mL of H.sub.2O was added 70 mg (1.75 mmol) of NaOH.
After stirring at 50.degree. C. for 16 hr, the solution was
concentrated to 1/3 volume then added dropwise to a stirred
solution of 10 mL of 0.5 M HCl(aq). After sitting at ambient
temperature for 5 min, the resulting solids were collected by
suction fitration, washed with H.sub.2O and dried to give 74 mg
(79%) of
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-quinolinecarboxylic acid as a yellow solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6): .delta. 9.01 (d, J=4 Hz, 1H), 8.73 (d, J=9
Hz, 1H), 8.25 (s, 1H), 7.99 (d, J=9 Hz, 1H), 7.87 (d, J=4 Hz, 1H),
7.74 (d, J=8 Hz, 2H), 7.62 (d, J=8 Hz, 2H), 7.55-7.51 (m, 1H), 6.93
(d, J=9 Hz, 2H), 4.87 (s, 2H), 3.45 (septet, J=7 Hz, 1H), 1.33 (d,
J=7, 6H). ESI-LCMS m/z 534 (M+H).sup.+.
Example 5
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-3-isoquinolinecarboxylic acid
##STR00092##
[0526] 5a) Methyl 7-hydroxy-3-isoquinolinecarboxylate
##STR00093##
[0528] To a solution of 4.36 g (22.6 mmol) of
7-hydroxy-1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid in 100
mL of MeOH was added 4.95 mL (67.7 mmol) of thionyl chloride and
the solution was refluxed for 1 hr. After evaporation of the
solvent, the residue was stirred vigorously in a mixture of 200 mL
of EtOAc, 500 mL of H.sub.2O and 100 mL of saturated
Na.sub.2CO.sub.3 (aq) until all solids dissolved. The aqueous layer
was then saturated with sodium chloride and extracted twice with
EtOAc. The combined organics were then washed with brine, dried
over Na.sub.2SO.sub.4 and concentrated to a solid. To this solid
was added 200 mL of xylenes and 3.0 g of palladium on carbon (10%)
and the suspension was refluxed for 16 hr. The suspension was
filtered through a pad of Celite.RTM. and silica gel. The pad was
then washed with 50 mL of xylenes. The filtrate was discarded and
the filter cake washed with 500 mL of a 4:1 mixture of
CH.sub.2Cl.sub.2:MeOH. This filtrate was then evaporated to give
2.09 g (46%) of methyl 7-hydroxy-3-isoquinolinecarboxylate as a
light brown solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
10.58 (s, 1H), 9.08 (s, 1H), 8.49 (s, 1H), 8.04 (d, J=9 Hz, 1H),
7.39 (d, J=9 Hz, 1H), 7.36 (s, 1H), 3.87 (s, 3H). ESI-LCMS m/z 204
(M+H).sup.+.
5b) Methyl
7-{[trifluoromethyl)sulfonyl]oxy}-3-isoquinolinecarboxylate
##STR00094##
[0530] To 2.09 g (10.3 mmol) of methyl
7-hydroxy-3-isoquinolinecarboxylate in 50 mL of CH.sub.2Cl.sub.2 at
0.degree. C. was added 3.60 mL (25.7 mmol) of TEA and 2.08 mL (12.3
mmol) of trifluoromethanesulfonic anhydride and the solution
stirred at ambient temperature for 1.5 hr. The solvent was
evaporated and the residue taken up in EtOAc. The organics were
washed with H.sub.2O, then brine and dried over Na.sub.2SO.sub.4.
The solvent was evaporated and the residue purified by silica gel
chromatography (120 g of silica gel eluting with 0-70% EtOAc in
hexanes over 45 minutes) to give 1.48 g of methyl
7-{[(trifluoromethyl)sulfonyl]oxy}-3-isoquinolinecarboxylate as a
beige solid. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.40 (s,
1H), 8.65 (s, 1H), 8.11 (d, J=9 Hz, 1H), 7.99 (d, J=2 Hz, 1H), 7.69
(dd, J.sub.A=2 Hz, J.sub.B=9 Hz, 1H), 4.07 (s, 3H). APCI-LCMS m/z
336 (M+H).sup.+.
Alternate 5b) Methyl 7-{[(trifluoromethyl)sulfonyl]oxy}-3-10
isoquinolinecarboxylate
##STR00095##
[0532] To 24.05 g (118 mmol) of methyl
7-hydroxy-3-isoquinolinecarboxylate, from multiple batches, in 500
mL of CH.sub.2Cl.sub.2 at 0.degree. C. was added 41.2 mL (296 mmol)
of TEA and 23.9 mL (142 mmol) of trifluoromethanesulfonic anhydride
and the solution stirred at ambient temperature for 20 minutes. The
solvent was evaporated and the residue taken up in EtOAc. The
organics were washed with H.sub.2O, then brine and dried over
Na.sub.2SO.sub.4. The mixture was filtered and the filtrate
concentrated to give methyl
7-{[(trifluoromethyl)sulfonyl]oxy}-3-isoquinolinecarboxylate (38.06
g, 96%) as a dark brown solid. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 9.40 (s, 1H), 8.65 (s, 1H), 8.11 (d, J=9 Hz, 1H), 7.99 (d,
J=2 Hz, 1H), 7.69 (dd, J.sub.A=2 Hz, J.sub.B=9 Hz, 1H), 4.07 (s,
3H). APCI-LCMS m/z 336 (M+H).sup.+.
5c) Methyl 7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate
##STR00096##
[0534] A suspension of 700 mg (2.09 mmol) of methyl
7-{[(trifluoromethyl)sulfonyl]oxy}-3-isoquinolinecarboxylate, 715
mg (3.13 mmol) of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol, 240 mg (0.21
mmol) of tetrakistriphenylphosphine palladium (0) and 3.70 mL (3.31
mmol) of 2.0 M Na.sub.2CO.sub.3 (aq) in 10 mL of
1,2-dimethoxyethane was stirred at 90.degree. C. for 20 min. The
suspension was filtered through a pad of Celite.RTM. and the filter
cake was washed with EtOAc. The combined organics were then
evaporated and the residue purified by silica gel chromatography
(40 g of silica gel eluting with 0-15% acetone in CH.sub.2Cl.sub.2
over 45 minutes) to give 360 mg (62%) of methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate as a beige solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 9.75 (s, 1H), 9.38 (s,
1H), 8.42 (s, 1H), 8.23 (s, 1H), 8.21 (d, J=8 Hz, 1H), 8.15 (d, J=8
Hz, 1H), 7.72 (d, J=9 Hz, 2H), 6.91 (d, J=9 Hz, 2H), 3.91 (s, 3H).
ESI-LCMS m/z 280 (M+H).sup.+.
Alternate 5c) Methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate
##STR00097##
[0536] A suspension of 38 g (113 mmol) of methyl
7-{[(trifluoromethyl)sulfonyl]oxy}-3-isoquinolinecarboxylate, 23.4
g (170 mmol) of (4-hydroxyphenyl)boronic acid, 6.54 g (5.67 mmol)
of tetrakistriphenylphosphine palladium (0) and 200 mL of 2.0 M
Na.sub.2CO.sub.3 (aq) in 700 mL of 1,2-dimethoxyethane was stirred
at 50.degree. C. for 90 min. The suspension was filtered through a
pad of Celite.RTM. and the filter cake was washed with EtOAc. The
combined organics were washed with water. The aqueous layer was
extracted with ethyl acetate. The combined organic layers were
washed with water and brine before being dried with
Na.sub.2SO.sub.4, filtered and concentrated. Ethyl acetate was
added and the mixture was stirred and heated. The mixture was
filtered and the concentrated filtrate was purified by silica gel
chromatography (eluting with 0-20% acetone in CH.sub.2Cl.sub.2).
Fractions containing the product were combined and concentrated and
combined with the solid which had been collected by filtration to
give 14.50 g (46%) of methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate. .sup.1H NMR (400
MHz, DMSO-d.sub.6): .delta. 9.75 (s, 1H), 9.38 (s, 1H), 8.42 (s,
1H), 8.23 (s, 1H), 8.21 (d, J=8 Hz, 1H), 8.15 (d, J=8 Hz, 1H), 7.72
(d, J=9 Hz, 2H), 6.91 (d, J=9 Hz, 2H), 3.91 (s, 3H). ESI-LCMS m/z
280 (M+H).sup.+.
5d) Methyl
7-[4-({[3-(2-chlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-3-isoquinolinecarboxylate
##STR00098##
[0538] A mixture of 174 mg (0.62 mmol) of methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate, 285 mg (0.92 mmol)
of
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
and 260 mg (1.83 mmol) of K.sub.2CO.sub.3 in 2 mL of DMF was
stirred at 50.degree. C. for 20 hr. EtOAc was added then the
mixture washed with three portions of H.sub.2O, then brine. The
organics were dried over Na.sub.2SO.sub.4 then evaporated and the
residue purified by silica gel chromatography (40 g of silica gel
eluting with 0-70% EtOAc in hexanes over 45 minutes) to give 196 mg
(58%) of methyl
7-[4-({[3-(2-chlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylate as white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 9.35 (s, 1H), 8.60 (s, 1H), 8.14 (s, 1H), 7.99
(m, 2H), 7.60 (d, J=9 Hz, 2H), 7.41 (d, J=9 Hz, 2H), 7.33 (m, 1H),
6.92 (d, J=9 Hz, 2H), 4.80 (s, 2H), 4.06 (s, 3H), 3.36 (septet, 1H,
J=7 Hz), 1.44 (d, J=7 Hz, 6H). ESI-LCMS m/z 548 (M+H).sup.+.
Alternate 5d) Methyl
7-[4-({[3-(2-chlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylate
##STR00099##
[0540] A mixture of 14.5 g (51.9 mmol) of methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate, 31.6 g (104 mmol) of
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
and 21.5 g (156 mmol) of K.sub.2CO.sub.3 in 100 mL of DMF was
stirred at 50.degree. C. for 20 hr. Additional
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(15.8 g, 52 mmol), DMF (50 mL), and K.sub.2CO.sub.3 (14.5 g, 105
mmol) was added and the reaction was heated for an additional 7
hours at 70.degree. C. and at 60.degree. C. for another 12 hours.
The reaction was allowed to cool to room temperature and was
stirred for 2 days. EtOAc was added then the mixture washed with
three portions of H.sub.2O, then brine. The aqueous layers were
extracted with EtOAC. The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered, then concentrated. The crude material
was dissolved in hot EtOAc and diluted with hexane. The mixture was
placed in the freezer for three hours then filtered to yield the
desired product (14.46 g) as a light brown solid. The filtrate was
concentrated and the residue purified by silica gel chromatography
(330 g of silica gel eluting with 0-80% EtOAc in hexanes over 45
minutes). Fractions containing the product were combined,
concentrated, and recrystallized to give 16.82 g (59%) of methyl
7-[4-({[3-(2-chlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylate. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 9.35 (s, 1H), 8.60 (s, 1H), 8.14 (s, 1H), 7.99 (m, 2H),
7.60 (d, J=9 Hz, 2H), 7.41 (d, J=9 Hz, 2H), 7.33 (m, 1H), 6.92 (d,
J=9 Hz, 2H), 4.80 (s, 2H), 4.06 (s, 3H), 3.36 (septet, 1H, J=7 Hz),
1.44 (d, J=7 Hz, 6H). ESI-LCMS m/z 548 (M+H).sup.+.
5e)
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}o-
xy)phenyl]-3-isoquinolinecarboxylic acid
##STR00100##
[0542] A solution of 16.4 g (30 mmol) of methyl
7-[4-({[3-(2-chlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylate and 11.98 g (300 mmol) of NaOH in a
mixture of 250 mL of EtOH, 100 mL of THF and 75 mL of H.sub.2O was
stirred at 50.degree. C. for 16 hr. The solution was concentrated
to 2/3 volume then added dropwise to a vigorously stirred solution
of 1.5 L of 1.0 N HCl (aq). After stirring for 30 min, the solids
were collected by suction filtration, washed with H.sub.2O then
dried to give 15.5 g (96%) of
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid as a pale yellow solid.
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 9.31 (s, 1H), 8.68 (s,
1H), 8.21 (s, 1H), 8.09-8.06 (m, 2H), 7.60 (d, J=9 Hz, 2H), 7.42
(d, J=8 Hz, 2H), 7.35-7.31 (m, 1H), 6.93 (d, J=9 Hz, 2H), 4.81 (d,
2H), 3.36 (septet, J=7 Hz, 1H), 1.44 (d, J=7 Hz, 6H). APCI-LCMS m/z
534 (M+H).sup.-.
5f) Sodium
7-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-3-isoquinolinecarboxylic acid
##STR00101##
[0544] A solution of 15.5 g (29.1 mmol) of
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid in 350 mL of MeOH was heated
to effect solution then 14.6 mL (29.1 mmol) of 2.0 M NaOH (aq) was
added. The solution was then concentrated and the solids dried to
yield 14.49 g (90%) of sodium
7-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid as a tan solid. .sup.1H NMR
(400 MHz, DMSO-d.sub.6): .delta. 9.35 (s, 1H), 8.56 (s, 1H), 8.42
(s, 1H), 8.17 (d, J=9 Hz, 1H), 8.12 (d, J=9 Hz, 1H), 7.74 (d, J=9
Hz, 2H), 7.62 (d, J=8 Hz, 2H), 7.55-7.51 (m, 1H), 6.93 (d, J=8 Hz,
2H), 4.88 (s, 2H), 3.45 (septet, J=7 Hz, 1H), 1.33 (d, J=7 Hz, 6H).
ESI-LCMS m/z 534 (M+H).sup.+.
Example 6
6-[4-({[5-Cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylic acid
##STR00102##
[0545] 6a) Ethyl 3-cyclopentyl-3-oxopropanoate
##STR00103##
[0547] To a solution of monoethyl malonate (13.6 mL, 115 mmol) and
a few milligrams of 2,2'-bipyridyl at between -55 and -65.degree.
C. was slowly added a 2.4 M solution of n-butyl lithium in hexanes
(92.0 mL, 230 mmol). After the addition was complete,
cyclopentanecarbonyl chloride (7.0 mL, 58 mmol) was added in
portions. The solution was then allowed to stir while warming to
ambient temperature and was poured into a mix of 1 N aqueous
hydrochloric acid and ether. The layers were separated and the
ether layer was washed three times with saturated sodium
bicarbonate, dried over magnesium sulfate, concentrated and
purified by chromatography (silica gel, 0-5% ethyl acetate in
hexanes gradient elution) to afford ethyl
3-cyclopentyl-3-oxopropanoate (9.50 g, 89%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.06 (q, J=7 Hz, 2H), 3.60 (s, 2H), 2.99-2.91
(m, 1H), 1.78-1.47 (m, 8H), 1.15 (t, J=7 Hz, 3H).
6b) Ethyl
5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate
##STR00104##
[0549] To a solution of 2,6-dichlorobenzaldehyde oxime (3.73 g,
19.6 mmol) in N,N-dimethylformamide (12 mL) was added solid
N-chlorosuccinimide (2.62 g, 19.6 mmol). The solution was stirred
for approximately 1 hour and the poured into water and extracted
twice with ether. The combined organic layers containing the crude
imidoyl chloride were dried over magnesium sulfate and
concentrated. To a solution of ethyl 3-cyclopentyl-3-oxopropanoate
(4.34 g, 23.6 mmol) in tetrahydrofuran (5 mL) at 0.degree. C. was
added a 25 wt % solution of sodium ethoxide in ethanol (7.4 mL, 24
mmol) quickly. The above imidoyl chloride was added. A solid was
seen to precipitate. The mixture was allowed to warm to ambient
temperature and stir overnight. The mixture was then poured into
water and extracted three times with ethyl acetate and the combined
organic layers were dried over magnesium sulfate, concentrated and
the residue purified by chromatography (silica gel 5% ethyl acetate
in hexanes) to afford ethyl
5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate (3.04
g, 43%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.63-7.53 (m,
3H), 4.02 (q, J=7 Hz, 2H), 3.92-3.84 (m, 1H), 2.14-2.06 (m, 2H),
1.84-1.64 (m, 6H), 0.91 (t, J=5 Hz, 3H).
6c) [5-Cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol
##STR00105##
[0551] To a solution of ethyl
5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate (3.0 g,
8.6 mmol) in tetrahydrofuran (35 mL) at 0.degree. C. was added a
1.5 M solution of diisobutylaluminum hydride in toluene (8.6 mL, 13
mmol) dropwise. The solution was allowed to stir while warming
slowly for approximately 3.5 hours and was re-cooled to 0.degree.
C. An additional portion of diisobutylaluminum hydride (4.3 mL, 6.5
mmol) was added and the solution was allowed to stir for an
additional 35 minutes. Then Rochelle's salt (40 mL) was added
slowly at 0.degree. C. followed by some ethyl acetate. The mixture
was allowed to warm to ambient temperature and stirred overnight.
The layers were separated and the ethyl acetate was washed with
brine, dried over magnesium sulfate and concentrated to afford
[5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol (2.59
g, 82%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.61-7.51 (m,
3H), 4.85 (t, J=5 Hz, 1H), 4.14 (d, J=5 Hz, 2H), 3.46-3.48 (m, 1H),
2.07-1.99 (m, 2H), 1.81-1.60 (m, 6H).
6d) Methyl
6-[4-({[5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate
##STR00106##
[0553] To a solution of methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (0.10 mg, 0.36 mmol),
[5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol (0.11
mg, 0.36 mmol) and triphenylphosphine (0.10 mg, 0.39 mmol) in
dichloromethane (2.5 mL) was added diisopropyl azodicarboxylate
(0.071 mL, 0.39 mmol) slowly. The solution was heated in a
microwave reactor at 90.degree. C. for 10 minutes and then was
adsorbed onto silica gel and purified by chromatography (silica
gel, 0-45% ethyl acetate in hexanes gradient elution) to afford
methyl
6-[4-({[5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylate (76 mg, 37%). .sup.1H-NMR (400 MHz, DMSO
d.sub.6) .delta. 8.55 (d, J=8 Hz, 1H), 8.28 (s, 1H), 8.18-8.10 (m,
3H), 7.73 (d, J=9 Hz, 2H), 7.63-7.51 (m, 3H), 6.96 (d, J=9 Hz, 2H),
4.98 (s, 2H), 3.93 (s, 3H), 3.58-3.50 (m, 1H), 2.11-2.03 (m, 2H),
1.84-1.64 (m, 6H).
6e)
6-[4-({[5-Cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-2-quinolinecarboxylic acid
##STR00107##
[0555] To a solution of methyl
6-[4-({[5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylate (73 mg, 0.13 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1N sodium hydroxide
(0.19 mL, 0.19 mmol). A solid was seen to precipitate. The mixture
was heated in a microwave reactor at 120.degree. C. for 500 seconds
and then was concentrated and the residue was taken up with water
and hydrochloric acid (0.19 mL, 0.19 mmol) was added. Ethyl acetate
was added and the layers were separated. The aqueous layer was
extracted once more with ethyl acetate. The combined organic layers
were washed with brine and concentrated to afford
6-[4-({[5-cyclopentyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylic acid (58 mg, 82%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 13.36 (br s, 1H), 8.53 (d, J=9 Hz, 1H), 8.27
(s, 1H), 8.18-8.08 (m, 3H), 7.73 (d, J=9 Hz, 2H), 7.63-7.51 (m,
3H), 6.92 (d, J=9 Hz, 2H), 4.87 (s, 2H), 3.59-3.51 (m, 1H),
2.11-2.03 (m, 2H), 1.84-1.64 (m, 6H). HRMS (ESI)
C.sub.31H.sub.24Cl.sub.2N.sub.2O.sub.4 calculated: 559.1186
(M+H).sup.+, found: 559.1183 (M+H).sup.+.
Example 7
6-[4-({[5-Cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phenyl-
]-2-quinolinecarboxylic acid
##STR00108##
[0556] 7a) Ethyl
5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate
##STR00109##
[0558] To a water bath-cooled solution of 2,6-dichlorobenzaldehyde
oxime (2.20 g, 11.6 mmol) in N,N-dimethylformamide (7 mL) was added
solid N-chlorosuccinimide (1.55 g, 11.6 mmol). The solution was
stirred while in the water bath for approximately 20 min and
outside the bath for approximately 1 hr. The solution was poured
into water and extracted twice with ether. The combined organic
layers containing the crude imidoyl chloride were dried over
magnesium sulfate and then concentrated. To a separate solution of
ethyl 3-cyclobutyl-3-oxopropanoate (2.37 g, 13.9 mmol) in THF (3
mL) at 0.degree. C. was added sodium ethoxide (25 wt % in ethanol,
4.36 mL, 13.9 mmol). The solution was stirred for a few minutes and
then the above imidoyl chloride was added. The solution was stirred
at 0.degree. C. for 10 minutes and then at ambient temperature
overnight. The solution was poured into water and extracted three
times with ethyl acetate. The combined organic layers were dried
over magnesium sulfate, concentrated and purified by chromatography
(silica gel, 0-5% ethyl acetate in hexanes gradient elution) to
afford ethyl
5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate (1.52 g,
38%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.63-7.53 (m,
3H), 4.29-4.20 (m, 1H), 4.02 (q, J=7 Hz, 2H), 2.43-2.36 (m, 4H),
2.15-2.06 (m, 1H), 1.97-1.89 (m, 1H), 0.94 (t, J=7 Hz, 3H).
7b) [5-Cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol
##STR00110##
[0560] To a solution of ethyl
5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate (1.52 g,
4.47 mmol) in THF (20 mL) at 0.degree. C. was slowly added a 1.5 M
solution of diisobutylaluminum hydride in toluene (6.26 mL, 9.39
mmol). The solution was allowed to warm slowly to ambient
temperature. After approximately 4 hr of stirring, the solution was
re-cooled to 0.degree. C. and an additional 6 mL of
diisobutylaluminum hydride was added. After approximately 1.5 hr of
stirring the mixture was re-cooled to 0.degree. C., quenched with
aqueous Rochelle's salt and allowed to stir overnight while warming
to ambient temperature. The mixture was extracted twice with ethyl
acetate. The combined organic layers were washed with Rochelle's
salt followed by with brine and then dried over magnesium sulfate
and concentrated to afford
[5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol (1.30 g,
98%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.61-7.51 (m,
3H), 4.87 (t, J=5 Hz, 1H), 4.10 (d, J=5 Hz, 2H), 3.91-3.83 (m, 1H),
2.42-2.28 (m, 4H), 2.08-1.98 (m, 1H), 1.94-1.84 (m, 1H).
7c) Methyl
6-[4-({[5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl-
}oxy)phenyl]-2-quinolinecarboxylate
##STR00111##
[0562] To a solution of
[5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol (0.12 g,
0.36 mmol), methyl 6-(4-hydroxyphenyl)-2-quinolinecarboxylate (0.10
g, 0.36 mmol) and triphenylphosphine (0.10 mg, 0.39 mmol) in
dichloromethane (2.5 mL) was added diisopropyl azodicarboxylate
(0.071 mL, 0.39 mmol) dropwise. The solution was heated in a
microwave reactor to 90.degree. C. for 10 minutes and the allowed
to sit overnight. The solution was adsorbed onto silica gel and
purified by chromatography (silica gel, 0-45% ethyl acetate in
hexanes gradient elution) to afford methyl
6-[4-({[5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylate (71 mg, 35%).
[0563] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (d, J=9 Hz,
1H), 8.28 (s, 1H), 8.16-8.10 (m, 3H), 7.73 (d, J=9 Hz, 2H),
7.62-7.51 (m, 3H), 6.92 (d, J=9 Hz, 2H), 4.84 (s, 2H), 4.05-3.97
(m, 1H), 3.93 (s, 3H), 2.43-2.31 (m, 4H), 2.14-2.02 (m, 1H),
1.97-1.89 (m, 1H). LRMS (APCI) m/z 559 (M+H).sup.+.
7d)
6-[4-({[5-Cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)ph-
enyl]-2-quinolinecarboxylic acid
##STR00112##
[0565] To a solution of methyl
6-[4-({[5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylate (67 mg, 0.12 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1 N sodium hydroxide
(0.18 mL, 0.18 mmol). The solution was heated in a microwave
reactor at 120.degree. C. for 500 seconds. The solution was
concentrated and water was added followed by 1 N hydrochloric acid
(0.18 mL, 0.18 mmol). Ethyl acetate was added. The layers were
separated and the aqueous layer was extracted with ethyl acetate
and the combined organic layers were washed with brine and
concentrated to afford of
6-[4-({[5-cyclobutyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylic acid as a solid (51 mg, 78%). .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 13.38 (br s, 1H), 8.52 (d, J=8 Hz,
1H), 8.27 (s, 1H), 8.15-8.08 (m, 3H), 7.73 (d, J=9 Hz, 2H),
7.62-7.51 (m, 3H), 6.92 (d, J=9 Hz, 2H), 4.84 (s, 2H), 4.07-3.94
(m, 1H), 2.43-2.33 (m, 4H), 2.11-2.03 (m, 1H), 1.94-1.90 (m,
1H).
[0566] HRMS (ESI) C.sub.30H.sub.22Cl.sub.2N.sub.2O.sub.4
calculated: 545.1029 (M+H).sup.+, found: 545.1026 (M+H).sup.+.
Example 8
6-4-({[5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-2-quinolinecarboxylic acid
##STR00113##
[0567] 8a) Methyl
6-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-2-quinolinecarboxylate
##STR00114##
[0569] To a solution of methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (0.10 mg, 0.36 mmol),
triphenylphosphine (94 mg, 0.36 mmol) and
[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methanol
(Example 9e) (0.11 mg, 0.36 mmol) in dichloromethane (2.5 mL) was
added diisopropyl azodicarboxylate (0.064 mL, 0.36 mmol). The
mixture was heated to 90.degree. C. for 10 minutes and then allowed
to stand overnight. The mixture was adsorbed onto silica gel and
purified by chromatography (silica gel, 0-40% ethyl acetate in
hexanes gradient elution followed by further purification (silica
gel, 0-30% ethyl acetate in hexanes gradient elution) to afford
methyl
6-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-2-quinolinecarboxylate (53 mg, 26%). .sup.1H-NMR (400 MHz,
DMSO-d6) .delta. 8.81 (s, 2H), 8.55 (d, J=9 Hz, 1H), 8.28 (s, 1H),
8.18-8.10 (m, 3H), 7.73 (d, J=9 Hz, 2H), 6.88 (d, J=9 Hz, 2H), 4.93
(s, 2H), 4.08-3.99 (m, 1H), 3.94 (s, 3H), 2.45-2.34 (m, 4H),
2.15-2.02 (m, 1H), 1.99-1.89 (m, 1H).
8b)
6-[4-({[5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylic acid
##STR00115##
[0571] To a solution of methyl
6-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-2-quinolinecarboxylate (53 mg, 0.095 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1 N sodium hydroxide
(0.14 mL, 0.14 mmol). The mixture was heated in a microwave reactor
at 120.degree. C. for 500 seconds and 1 N hydrochloric acid (0.14
mL, 0.14 mmol) was added. The mixture was concentrated, water was
added and the mixture was extracted with ethyl acetate. The aqueous
layer was extracted one more time with ethyl acetate. The combined
organic layers were washed with brine and concentrated. The solid
was washed with ether and dried overnight under a nitrogen stream
at 45.degree. C. to afford
6-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-2-quinolinecarboxylic acid as a yellow solid (27 mg, 52%).
.sup.1H-NMR (400 MHz, DMSO-d6) .delta. 8.81 (s, 2H), 8.53 (d, J=9
Hz, 1H), 8.27 (s, 1H), 8.17-8.08 (m, 3H), 7.73 (d, J=9 Hz, 2H),
6.88 (d, J=9 Hz, 2H), 4.93 (s, 2H), 4.08-3.97 (m, 1H), 2.44-2.33
(m, 4H), 2.14-2.04 (m, 1H), 1.99-1.90 (m, 1H). HRMS (ESI)
C.sub.29H.sub.21Cl.sub.2N.sub.3O.sub.4: 546.0982 (M+H).sup.+,
found: 546.0982 (M+H).sup.+.
Example 9
7-[4-({[5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid
##STR00116##
[0572] 9a) Ethyl 3-cyclobutyl-3-oxopropanoate
##STR00117##
[0574] To a solution of monoethyl malonate (12.0 mL, 101 mmol) and
a few milligrams of 2,2'-bipyridyl in tetrahydrofuran (250 mL) at
approximately -55.degree. C. was added in a dropwise fashion butyl
lithium (2.5 M in hexanes, 81.0 mL, 202 mmol). Then
cyclobutanecarbonyl chloride (6.00 mL, 50.6 mmol) was added
dropwise. The flask was removed from the cold bath and the mixture
was allowed to stir while warming to ambient temperature. The
mixture was poured into 1 N aqueous hydrochloric acid and extracted
twice with ether. The combined organic layers were dried over
magnesium sulfate, washed twice with saturated sodium bicarbonate,
dried again over magnesium sulfate, concentrated and purified by
chromatography (silica gel, 7.5% ethyl acetate in hexanes) to
afford ethyl 3-cyclobutyl-3-oxopropanoate (6.69 g, 78%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 4.06 (q, J=7 Hz, 2H),
3.49 (s, 2H), 3.38-3.32 (m, 1H), 2.11-2.05 (m, 4H), 1.90-1.85 (m,
1H), 1.87-1.66 (m, 1H) 1.15 (t, J=7 Hz, 3H).
9b) 3,5-Dichloro-4-pyridinecarbaldehyde oxime
##STR00118##
[0576] To a suspension of 3,5-dichloro-4-pyridinecarbaldehyde (6.50
g, 36.9 mmol) in ethanol (55 mL) was added a solution of sodium
hydroxide (1.77 g, 44.3 mmol) and hydroxylamine hydrochloride (3.08
g, 44.3 mmol) in water (30 mL). The suspension was heated to
90.degree. C. The mixture was stirred at 90.degree. C. for
approximately 6.5 hours and then concentrated. Water was added and
the resulting solid was isolated by filtration and dried under a
nitrogen stream at 45.degree. C. over phosphorus pentoxide to
afford 3,5-dichloro-4-pyridinecarbaldehyde oxime (6.38 g, 33.4
mmol, 91%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 13.31 (s,
1H), 8.68 (s, 2H), 8.24 (s, 1H).
9c) Ethyl
5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylate
##STR00119##
[0578] To a solution of 3,5-dichloro-4-pyridinecarbaldehyde oxime
(3.15 g, 16.5 mmol) in N,N-dimethylformamide (13 mL) was added
N-chlorosuccinimide (2.20 g, 16.5 mmol). The mixture was heated to
65.degree. C. and all solids dissolved. The solution was stirred at
65.degree. C. for approximately 1.5 hours, poured into water and
extracted with ether. The ether layer containing the crude imidoyl
chloride was washed with brine, dried over magnesium sulfate and
concentrated. To a separate solution of ethyl
3-cyclobutyl-3-oxopropanoate (3.37 g, 19.8 mmol) in tetrahydrofuran
(4 mL) at 0.degree. C. was added sodium ethoxide (25 wt % in
ethanol, 6.21 mL, 19.8 mmol) and the mixture was stirred for
approximately 7 minutes. Then the above imidoyl chloride was added
in tetrahydrofuran (13 mL) at a slow rate. A solid precipitated and
the mixture was allowed to warm to ambient temperature and stir
overnight. The mixture was concentrated and taken up with ethyl
acetate and washed with water. The aqueous layer was back-extracted
with ethyl acetate and the combined organic layers were washed with
brine, dried over magnesium sulfate and purified by chromatography
(silica gel, 0-10% ethyl acetate in hexanes gradient elution) to
afford ethyl
5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylate
(2.98 g, 53%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 8.64 (s,
2H), 4.35-4.20 (m, 1H), 4.05 (q, J=7 Hz, 2H), 2.45-2.30 (m, 4H),
2.14-2.04 (m, 1H), 2.00-1.89 (m, 1H), 0.96 (t, J=7 Hz, 3H). LRMS
(APCI) m/z 341 (M+H).sup.+.
9d) 5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylic
acid
##STR00120##
[0580] To a solution of ethyl
5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylate
(1.00 g, 2.93 mmol), in 2:1 tetrahydrofuran:methanol (15 mL) was
added 1 N aqueous sodium hydroxide (4.40 mL, 4.40 mmol). The
solution was heated in a microwave reactor to 120.degree. C. for
500 seconds. Then 1 N aqueous hydrochloric acid (4.40 mL, 4.40
mmol) was added and the solution was concentrated. Water was added
and the mixture was extracted twice with ethyl acetate. The
combined organic layers were washed with brine, dried over
magnesium sulfate, concentrated and dried under a nitrogen stream
at 45.degree. C. overnight to afford
5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylic
acid (808 mg, 88%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
13.34 (br s, 1H), 8.81 (s, 2H), 4.33-4.25 (m, 1H), 2.42-2.32 (m,
4H), 2.15-2.05 (m, 1H), 1.96-1.88 (m, 1H).
9e)
[5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methanol
##STR00121##
[0582] To a solution of
5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylic
acid (808 mg, 2.59 mmol) in THF (13 mL) at 0.degree. C. was added
triethylamine (0.361 mL, 2.59 mmol) followed by isopropyl
chloroformate (1 M in toluene, 2.59 mL, 2.59 mmol). The solution
was stirred at 0.degree. C. for approximately 1 hour and then was
filtered into a solution of sodium borohydride (127 mg, 3.37 mmol)
in water (1 mL) at 0.degree. C. The solution was allowed to warm to
ambient temperature and was then concentrated. Water was added and
the mixture was extracted three times with ethyl acetate. The
combined organic layers were washed with brine, dried over
magnesium sulfate, concentrated and purified by chromatography
(silica gel, 0-40% ethyl acetate in hexanes gradient elution) to
afford
[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methanol
(478 mg, 62%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 8.79 (s,
2H), 4.97-4.94 (m, 1H), 4.16-4.15 (m, 2H), 3.92-3.83 (m, 1H),
2.40-2.30 (m, 4H), 2.10-2.01 (m, 1H), 1.99-1.89 (m, 1H).
9f) Methyl
7-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]-
methyl}oxy)phenyl]-3-isoquinolinecarboxylate
##STR00122##
[0584] To a solution of methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate (50 mg, 0.18 mmol),
[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methanol
(54 mg, 0.18 mmol) and triphenylphosphine (52 mg, 0.20 mmol) in
dichloromethane (1.5 mL) was added diisopropyl azodicarboxylate
(0.035 mL, 0.20 mmol). The solution was heated in a microwave
reactor at 90.degree. C. for 10 minutes and allowed to stand at
ambient temperature overnight. The next day the solution was
adsorbed onto silica gel and purified by chromatography (first in
0-1.75% methanol in chloroform gradient elution then in 0-2%
methanol in chloroform) to afford methyl
7-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-3-isoquinolinecarboxylate as a white solid (39 mg, 39%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 9.33 (s, 1H), 8.59 (s,
3H), 8.12 (s, 1H), 8.00-7.94 (m, 2H), 7.58 (d, J=9 Hz, 2H), 6.84
(d, J=9 Hz, 2H), 4.79 (s, 2H), 4.04 (s, 3H), 3.87-3.79 (m, 1H),
2.64-2.54 (m, 2H), 2.50-2.35 (m, 2H), 2.19-2.00 (m, 2H).
9g)
7-[4-({[5-Cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-3-isoquinolinecarboxylic acid
##STR00123##
[0586] To a solution of methyl
7-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-3-isoquinolinecarboxylate (39 mg, 0.070 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1 N sodium hydroxide
(0.10 mL, 0.10 mmol). The solution was heated in a microwave
reactor at 120.degree. C. for 500 seconds. The solution was
concentrated under a nitrogen stream and 1 N aqueous hydrochloric
acid (0.10 mL, 0.10 mmol) was added. The solution was concentrated
further under an nitrogen stream and water was added followed by
ethyl acetate. A solid precipitated.
[0587] The water layer was removed and ether was added and the
solid was isolated by filtration. The solid was dried under a
nitrogen stream at 45.degree. C. to afford
7-[4-({[5-cyclobutyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-3-isoquinolinecarboxylic acid (23 mg, 60%). .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.39 (s, 1H), 8.81 (s, 2H), 8.60
(s, 1H), 8.43 (s, 1H), 8.25-8.10 (m, 2H), 7.75 (d, J=9 Hz, 2H),
6.89 (d, J=9 Hz, 2H), 4.93 (s, 2H), 4.08-3.99 (m, 1H), 2.50-2.40
(m, 4H), 2.15-2.05 (m, 1H), 2.00-1.90 (m, 1H).
[0588] HRMS (ESI) C.sub.29H.sub.21Cl.sub.2N.sub.3O.sub.4
calculated: 546.0982 [M+H].sup.+, found: 546.0981 [M+H].sup.+.
Example 10
7-[4-({[5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-3-isoquinolinecarboxylic acid
##STR00124##
[0589] 10a) Ethyl 3-cyclopropyl-3-oxopropanoate
##STR00125##
[0591] Ethyl 3-cyclopropyl-3-oxopropanoate was made by the same
procedure as the one employed to synthesize methyl
3-cyclobutyl-3-oxopropanoate in example 9a, except
cyclopropylcarbonyl chloride was used instead of cyclobutylcarbonyl
chloride, affording ethyl 3-cyclopropyl-3-oxopropanoate (3.35 g,
56%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 4.07 (q, J=7 Hz,
2H), 3.65 (s, 2H), 2.09-2.02 (m, 1H), 1.16 (t, J=7 Hz, 3H),
0.93-0.86 (m, 4H).
10b) Ethyl
5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate
##STR00126##
[0593] To a water bath-cooled solution of 2,6-dichlorobenzaldehyde
oxime (2.20 g, 11.6 mmol) in N,N-dimethylformamide (7 mL) was added
solid N-chlorosuccinimide (1.55 g, 11.6 mmol). The solutions were
stirred while in the water bath for approximately 20 min and the
outside the bath for approximately 1 hour. The solution was poured
into water and extracted twice with ether. The combined organic
layers containing the crude imidoyl chloride were dried over
magnesium sulfate and then concentrated. To a separate solution of
ethyl 3-cyclopropyl-3-oxopropanoate (2.17 g, 13.9 mmol) in THF (3
mL) at 0.degree. C. was added sodium ethoxide (25 wt % in ethanol,
4.36 mL, 13.9 mmol). The solution was stirred for a few minutes and
then the above imidoyl chloride was added. The solution was stirred
at 0.degree. C. for 10 minutes and then at ambient temperature
overnight. The solution was poured into water and extracted three
times with ethyl acetate. The combined organic layers were dried
over magnesium sulfate, concentrated and purified by chromatography
(silica gel, 0-5% ethyl acetate in hexanes gradient elution) to
afford ethyl
5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate (289
mg, 8%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.62-7.53 (m,
3H), 4.04 (q, J=7 Hz, 2H), 2.84-2.82 (m, 1H), 1.35-1.25 (m, 4H),
0.93 (t, J=7 Hz, 3H).
10c)
[5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol
##STR00127##
[0595] To a solution of ethyl
5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolecarboxylate (289
mg, 0.886 mmol) in THF (4 mL) at 0.degree. C. was slowly added
diisobutylaluminum hydride (1.5M in toluene, 1.24 mL, 1.86 mmol).
The solution was allowed to warm slowly to ambient temperature.
After approximately 4 hours of stirring, the solution was re-cooled
to 0.degree. C. and an additional 1 mL of diisobutylaluminum
hydride was added. After approximately 1.5 hours of stirring the
mixture was re-cooled to 0.degree. C., quenched with some aqueous
Rochelle's salt and allowed to stir overnight while warming to
ambient temperature. The mixture was extracted twice with ethyl
acetate. The combined organic layers were washed once with
Rochelle's Salt, dried over magnesium sulfate and concentrated to
afford [5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol
(242 mg, 96%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
7.61-7.51 (m, 3H), 4.93 (t, J=5 Hz, 1H), 4.19 (d, J=5 Hz, 2H),
2.31-2.27 (m, 1H), 1.11-1.04 (m, 4H).
10d) Methyl
7-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylate
##STR00128##
[0597] To a solution of
[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol (51 mg,
0.18 mmol), methyl 7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate
(50 mg, 0.18 mmol) and triphenylphosphine (52 mg, 0.20 mmol) in
dichloromethane (1.5 mL) was added diisopropyl azodicarboxylate
(0.035 mL, 0.20 mmol). The solution was heated in a microwave
reactor at 90.degree. C. for 10 minutes and then allowed to stand
overnight. The mixture was adsorbed onto silica gel and purified by
chromatography (silica gel, 0-1.25% methanol in dichloromethane) to
afford methyl
7-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl]methyl}o-
xy)phenyl]-3-isoquinolinecarboxylate (54 mg, 55%). .sup.1H-NMR (400
MHz, DMSO-d.sub.6) .delta. 9.39 (s, 1H), 8.63 (s, 1H), 8.44 (s,
1H), 8.29-8.14 (m, 2H), 7.75 (d, J=9 Hz, 2H), 7.61-7.50 (m, 3H),
6.96 (d, J=9 Hz, 2H), 4.94 (s, 2H), 3.91 (s, 3H), 2.50-2.46 (m,
1H), 1.21-1.14 (m, 4H).
10e)
7-[4-({[5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-isoquinolinecarboxylic acid
##STR00129##
[0599] To a solution of methyl
7-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylate (54 mg, 0.099 mmol) in 2:1
tetrahydrofuran:methanol (1.5 ml) was added 1 N sodium hydroxide
(0.15 mL, 0.15 mmol). The solution was heated in a microwave
reactor to 120.degree. C. for 500 seconds. The solution was
concentrated under a nitrogen stream and 1 N hydrochloric acid
(0.15 mL, 0.15 mmol) was added. The solution was further
concentrated under a nitrogen stream. Water and ethyl acetate were
added and a solid precipitated that would not dissolve. The water
layer was removed and the resulting mixture was washed twice with
water. The organic solvents were removed under vacuum and the
resulting solid was dried at 45.degree. C. under a nitrogen stream
to afford
7-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-3-isoquinolinecarboxylic acid (36 mg, 68%). .sup.1H-NMR (400
MHz, DMSO-d.sub.6) .delta. 13.1 (br s, 1H), 9.39 (s, 1H), 8.60 (s,
1H), 8.43 (s, 1H), 8.22-8.13 (m, 2H), 7.75 (d, J=9 Hz, 2H),
7.61-7.51 (m, 3H), 6.96 (d, J=9 Hz, 2H), 4.94 (s, 2H), 2.51-2.45
(m, 1H), 1.12-1.13 (m, 4H). HRMS (ESI)
C.sub.29H.sub.20Cl.sub.2N.sub.2O.sub.4 calculated 531.0880
(M+H).sup.+, found: 531.0872 (M+H).sup.+.
Example 11
6-[4-({[5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylic acid
##STR00130##
[0600] 11a) Methyl
6-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylate
##STR00131##
[0602] To a solution of
[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methanol (0.10
g, 0.36 mmol), methyl 6-(4-hydroxyphenyl)-2-quinolinecarboxylate
(0.10 g, 0.36 mmol) and triphenylphosphine (0.10 g, 0.39 mmol) in
dichloromethane (2.5 mL) was added diisopropyl azodicarboxylate
(0.071 mL, 0.39 mmol) dropwise. The solution was heated in a
microwave reactor to 90.degree. C. for 10 minutes and then allowed
to sit overnight. The solution was adsorbed onto silica gel and
purified by chromatography (silica gel, 0-45% ethyl acetate in
hexanes gradient elution) to afford methyl
6-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylate (62 mg, 32%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.55 (d, J=9 Hz, 1H), 8.28 (s, 1H), 8.18-8.10
(m, 3H), 7.73 (d, J=9 Hz, 2H), 7.62-7.50 (m, 3H), 6.95 (d, J=9 Hz,
2H), 4.93 (s, 2H), 3.93 (s, 3H), 2.50-2.42 (m, overlapping DMSO
.about.1H), 1.20-1.12 (m, 4H). LRMS (APCI) m/z 545 (M+H).sup.+.
11b)
6-[4-({[5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinolinecarboxylic acid
##STR00132##
[0604] To a solution of methyl
6-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylate (58 mg, 0.11 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1 N sodium hydroxide
(0.18 mL, 0.18 mmol). The solution was heated in a microwave
reactor at 120.degree. C. for 500 seconds. The solution was
concentrated and water was added followed by 1 N hydrochloric acid
(0.18 mL, 0.18 mmol). The solution was extracted twice with ethyl
acetate and the combined organic layers were washed with brine and
concentrated to afford
6-[4-({[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methyl}oxy)phen-
yl]-2-quinolinecarboxylic acid (52 mg, 92%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 13.36 (br s, 1H), 8.53 (d, J=9 Hz, 1H), 8.27
(s, 1H), 8.17-8.08 (m, 3H), 7.73 (d, J=9 Hz, 2H), 7.62-7.51 (m,
3H), 6.95 (d, J=9 Hz, 2H), 4.93 (s, 2H), 2.49-2.43 (m, 1H),
1.20-1.10 (m, 4H). HRMS (ESI)
C.sub.29H.sub.20Cl.sub.2N.sub.2O.sub.4 calculated: 531.0873
[M+H].sup.+, found: 531.0873 [M+H].sup.+.
Example 12
6-[4-({[5-Cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}oxy-
)phenyl]-2-quinolinecarboxylic acid
##STR00133##
[0605] 12a) Ethyl
5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylate
##STR00134##
[0607] To a solution of 3,5-dichloro-4-pyridinecarbaldehyde oxime
(1.44 g, 7.53 mmol) in N,N-dimethylformamide (6 mL) was added
N-chlorosuccinimide (1.00 g, 7.53 mmol). The mixture was heated to
65.degree. C. and all solids dissolved. The solution was stirred at
65.degree. C. for approximately 1.5 hr, poured into water and
extracted with ether. The ether layer containing the crude imidoyl
chloride was washed with brine, dried over magnesium sulfate and
concentrated. To a separate solution of ethyl
3-cyclopropyl-3-oxopropanoate (1.41 g, 9.03 mmol) in
tetrahydrofuran (2 mL) at 0.degree. C. was added a 25 wt % solution
of sodium ethoxide in ethanol (2.83 mL, 9.03 mmol) and the mixture
was stirred for approximately 7 min. Then the above imidoyl
chloride in tetrahydrofuran (6.5 mL) was added at a slow rate. A
solid precipitated and the mixture was allowed to warm to ambient
temperature and stir overnight. The mixture was concentrated and
taken up with ethyl acetate and washed with water. The aqueous
layer was back-extracted with ethyl acetate and the combined
organic layers were washed with brine, dried over magnesium sulfate
and purified by chromatography (silica gel, 0-10% ethyl acetate in
hexanes gradient elution). The isolated solid was washed with
hexanes to afford ethyl
5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylate
(214 mg, 9%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 8.84 (s,
2H), 4.07(q, J=7 Hz, 2H), 2.88-2.81 (m, 1H), 1.37-1.27 (m, 4H),
0.95 (t, J=7 Hz, 3H). LRMS (APCI) m/z 327 (M+H).sup.+.
12b)
5-Cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylic
acid
##STR00135##
[0609] To a solution of ethyl
5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylate
(210 mg, 0.642 mmol) in 2:1 tetrahydrofuran:methanol (3 mL) was
added 1 N sodium hydroxide (0.963 mL, 0.963 mmol). The solution was
heated in a microwave reactor to 120.degree. C. for 500 seconds.
Then 1 N hydrochloric acid (0.963 mL, 0.963 mmol) was added and the
solution was concentrated. The residue was taken up with water and
extracted twice with ethyl acetate. The combined organic layers
were washed with brine, dried over magnesium sulfate and
concentrated to afford
5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylic
acid (158 mg, 82%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
13.35 (s, 1H), 8.81 (s, 2H), 2.91-2.84 (m, 1H), 1.34-1.24 (m,
4H).
12c)
[5-Cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methanol
##STR00136##
[0611] To a solution of
5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolecarboxylic
acid (155 mg, 0.52 mmol) and triethylamine (0.072 mL, 0.52 mmol) in
tetrahydrofuran (3 mL) at 0.degree. C. was added isopropyl
chloroformate (0.52 mL, 0.52 mmol). A white solid was seen to
precipitate. The mixture was stirred at 0.degree. C. for
approximately 1 hour and then filtered into a stirring solution of
sodium borohydride (25 mg, 0.67 mmol) in water at 0.degree. C. The
solution was stirred at 0.degree. C. for approximately 30 minutes
and the allowed to warm to ambient temperature. The solution was
concentrated and water was added. The solution was extracted twice
with ethyl acetate. The combined organic layers were washed with
brine, dried over magnesium sulfate, concentrated and purified by
chromatography (silica gel, 0-40% ethyl acetate in hexanes gradient
elution) to afford
[5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methanol
(83 mg, 56%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.62 (s,
2H), 4.44 (s, 2H), 2.20-2.14 (m, 1H), 1.30-1.44 (m, 4H).
12d) Methyl
6-[4-({[5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}ox-
y)phenyl]-2-quinolinecarboxylate
##STR00137##
[0613] To a solution of methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (81 mg, 0.29 mmol),
triphenylphosphine (84 mg, 0.32 mmol) and
[5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methanol
(83 mg, 0.29 mmol) in dichloromethane (2.5 mL) was added
diisopropyl azodicarboxylate (0.058 mL, 0.32 mmol) dropwise. The
solution was heated in a microwave reactor at 90.degree. C. for 10
minutes and then allowed to stand overnight. The solution was
adsorbed onto silica gel and purified by chromatography (silica
gel, 0-1.25% methanol in chloroform gradient elution) to afford
methyl
6-[4-({[5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}ox-
y)phenyl]-2-quinolinecarboxylate (70 mg, 44%). .sup.1H-NMR (400
MHz, CDCl.sub.3) .delta. 8.60 (s, 2H), 8.39-8.33 (m, 2H), 8.22 (d,
J=9 Hz, 1H), 8.01-7.96 (m, 2H), 7.60 (d, J=9 Hz, 2H), 6.88 (d, J=9
Hz, 2H), 4.90 (s, 2H), 4.09 (s, 3H), 2.22-2.15 (m, 1H), 1.33-1.13
(m, 4H).
12e)
6-[4-({[5-Cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00138##
[0615] To a solution of methyl
6-[4-({[5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}ox-
y)phenyl]-2-quinolinecarboxylate (70 mg, 0.13 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1 N sodium hydroxide
(0.19 mL, 0.19 mmol). The solution was heated in a microwave
reactor at 120.degree. C. for 500 seconds. Then (0.19 mL, 0.19
mmol) 1 N hydrochloric acid was added and the solution was
concentrated. The residue was taken up with water and extracted
twice with ethyl acetate. The combined organic layers were washed
with brine and concentrated. The resulting solid was dried under a
nitrogen stream at 45.degree. C. over 48 hours to afford
6-[4-({[5-cyclopropyl-3-(3,5-dichloro-4-pyridinyl)-4-isoxazolyl]methyl}ox-
y)phenyl]-2-quinolinecarboxylic acid as a yellow solid (53 mg,
78%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 13.37 (br s, 1H),
8.81 (s, 2H), 8.53 (d, J=9 Hz, 1H), 8.28 (s, 1H), 8.18-8.08 (m,
3H), 7.74 (d, J=9 Hz, 2H), 6.92 (d, J=9 Hz, 2H), 5.02 (s, 2H),
2.52-2.46 (m, 1H), 1.23-1.10 (m, 4H). HRMS (ESI)
C.sub.28H.sub.19Cl.sub.2N.sub.3O.sub.4 calculated: 532.0825
[M+H].sup.+, found: 532.0823 [M+H].sup.+.
Example 13
6-{4-[({3-(2,6-Dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methyl-
)oxy]phenyl}-2-quinolinecarboxylic acid
##STR00139##
[0616] 13a) Ethyl 4-methyl-3-oxohexanoate
##STR00140##
[0618] To a solution of monoethyl malonate (49.5 mL, 0.42 mol) and
a few milligrams of 2,2'-bipyridyl in tetrahydrofuran at
-70.degree. C. was added 2.5 M n-butyl lithium in hexanes (0.34 L,
0.84 mol) dropwise over approximately 1.5 hours. Then
2-methylbutyryl chloride (26 mL, 0.21 mol) was added dropwise. The
mixture was allowed to warm to ambient temperature and was then
poured into 1 N hydrochloric acid. The mixture was extracted three
times with ether and the combined organic layers were dried over
magnesium sulfate and concentrated. The resulting oil was purified
by vacuum distillation (boiling range 75-85.degree. C.) to afford
ethyl 4-methyl-3-oxohexanoate (36.4 g, 100%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 4.06 (q, J=7 Hz, 2H), 3.60 (s, 2H), 2.57-2.49
(m, 1H), 1.64-1.53 (m, 1H), 1.36-1.26 (m, 1H), 1.15 (t, J=7 Hz,
3H), 0.97 (d, J=7 Hz, 3H), 0.78 (t, J=7 Hz, 3H).
13b)
{3-(2,6-Dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methanol
##STR00141##
[0620] To a solution of 2,6-dichlorobenzaldehyde oxime (26.4 g, 139
mmol) in N,N-dimethylformamide (70 mL) at 5.degree. C. was added
solid N-chlorosuccinimide (18.6 g, 139 mmol) in portions. The
mixture was allowed to stir and warm to ambient temperature (with
occasional cooling when warming was noted) over approximately 1.5
hour and then poured into ether. The ether layer containing the
crude imidoyl chloride was washed twice with water followed by
brine, dried over magnesium sulfate and concentrated. To a separate
solution of ethyl 4-methyl-3-oxohexanoate (28.8 g, 167 mmol) in
tetrahydrofuran (50 mL) at 0.degree. C. was added sodium ethoxide
(21 wt % in ethanol, 62.3 mL, 167 mmol) quickly. Then the above
imidoyl chloride was added dropwise in tetrahydrofuran (100 mL).
The solution was allowed to stir while warming to ambient
temperature overnight. The mixture was then poured into ether,
washed twice with water followed by brine, dried over magnesium
sulfate, concentrated and purified by chromatography (silica gel,
1.25% acetone in hexanes) to afford an oil which was dissolved in
tetrahydrofuran (75 mL) at 0.degree. C. Then a 1.5 M solution of
diisobutylaluminum hydride in toluene (100 mL, 150 mmol) was added
slowly. The solution was allowed to warm slightly while stirring
for approximately 2.5 hours and then was re-cooled to 0.degree. C.
and stirred for 1.5 hours. Then aqueous Rochelle's salt (20 mL) was
added carefully followed by ethyl acetate and the mixture was
stirred overnight. Then the layers were separated and the aqueous
layer was extracted two more times with ethyl acetate. The combined
organic layers were washed with brine, dried over magnesium
sulfate, concentrated and purified by chromatography (silica gel,
20% ethyl acetate in hexanes) to afford a white solid. The
enantiomers were resolved by supercritical fluid chromatography
(Chiralpak AD, 10% methanol in supercritical carbon dioxide) and
the resulting solid was taken up with dichloromethane, dried over
magnesium sulfate and concentrated to afford
{3-(2,6-dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methanol
(1.53 g, 7%) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.61-7.51
(m, 3H), 4.85 (t, J=5 Hz, 1H), 4.20-4.06 (m, 2H), 3.15-3.10 (m,
1H), 1.69-1.62 (m, 2H), 1.29 (d, J=7 Hz, 3H), 0.79 (t, J=7 Hz,
3H).
13c) Methyl
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylate
##STR00142##
[0622] To a solution of methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (154 mg, 0.550 mmol),
triphenylphosphine (144 mg, 0.550 mmol) and
{3-(2,6-dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methanol
(150 mg, 0.500 mmol) in dichloromethane (1.5 mL) was added
diisopropyl azodicarboxylate (0.099 mL, 0.550 mmol) dropwise. The
solution was heated in a microwave reactor at 90.degree. C. for 10
minutes. The solution was adsorbed onto silica gel and purified by
chromatography (silica gel, 0-60% ethyl acetate in hexanes gradient
elution). A second purification was done by chromatography (silica
gel, 2% methanol in dichloromethane) to afford methyl
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylate as a white solid (118 mg,
42%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (d, J=9 Hz,
1H), 8.28 (s, 1H), 8.18-8.10 (m, 3H), 7.73 (d, J=8 Hz, 2H),
7.63-7.51 (m, 3H), 6.92 (d, J=8 Hz, 2H), 4.88 (app q, J=12 Hz, 2H),
3.94 (s, 3H), 3.27-3.22 (m, 1H), 1.73-1.66 (m, 2H), 1.32 (d, J=7
Hz, 3H), 0.83 (t, J=7 Hz, 3H).
13d)
6-{4-[({3-(2,6-Dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazolyl}m-
ethyl)oxy]phenyl}-2-quinolinecarboxylic acid
##STR00143##
[0624] To a solution of methyl
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1S)-1-methylpropyl]-isoxazolyl}methyl)-
oxy]phenyl}-2-quinolinecarboxylate (50 mg, 0.089 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1 N sodium hydroxide
(0.13 mL, 0.13 mmol). The solution was heated in a microwave
reactor at 80.degree. C. for 10 minutes and then 1 N hydrochloric
acid (0.13 mL, 0.13 mmol) was added. The solution was concentrated
and the residue was taken up with ethyl acetate and water. The
layers were separated and the organic layer was dried over
magnesium sulfate and concentrated to afford 50 mg of
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1S)-1-methylpropyl]-4-isoxazo-
lyl}methyl)oxy]phenyl}-2-quinolinecarboxylic acid as a yellow solid
(0.089 mmol, 100%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
13.37 (br s, 1H), 8.53 (d, J=9 Hz, 1H), 8.27 (s, 1H), 8.18-8.08 (m,
3H), 7.73 (d, J=9 Hz, 2H), 7.63-7.51 (m, 3H), 6.99 (d, J=9 Hz, 2H),
4.88 (app q, J=12 Hz, 2H), 3.31-3.22 (m, 1H), 1.73-1.66 (m, 2H),
3.32 (d, J=7 Hz, 3H), 0.83 (t, J=7 Hz, 3H). HRMS (ESI)
C.sub.30H.sub.24Cl.sub.2N.sub.2O.sub.4 calculated: 547.1186
(M+H).sup.30 , found: 547.1182 (M+H).sup.+.
Example 14
6-{4-[({3-(2,6-Dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methyl-
)oxy]phenyl}-2-quinolinecarboxylic acid
##STR00144##
[0625] 14a)
{3-(2,6-Dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methanol
##STR00145##
[0627] To a solution of 2,6-dichlorobenzaldehyde oxime (26.4 g, 139
mmol) in DMF (70 mL) at 5.degree. C. was added solid
N-chlorosuccinimide (18.6 g, 139 mmol) in portions. The mixture was
allowed to stir and warm to ambient temperature (with occasional
cooling when warming was noted) over approximately 1.5 hour and
then poured into ether. The organic layer containing the crude
imidoyl chloride was washed twice with water followed by brine,
dried over magnesium sulfate and concentrated. To a separate
solution of ethyl 4-methyl-3-oxohexanoate (28.8 g, 167 mmol) in
tetrahydrofuran (50 mL) at 0.degree. C. was added a 21 wt %
solution of sodium ethoxide in ethanol (62.3 mL, 167 mmol) quickly.
The above imidoyl chloride was added dropwise in tetrahydrofuran
(100 mL). The solution was allowed to stir while warming to ambient
temperature overnight. The mixture was then poured into ether,
washed twice with water followed by brine, dried over magnesium
sulfate, concentrated and purified by chromatography (silica gel,
1.25% acetone in hexanes) to afford an oil which was dissolved in
tetrahydrofuran (75 mL) at 0.degree. C. Then a 1.5 M solution of
diisobutylaluminum hydride in toluene (100 mL, 150 mmol) was added
slowly. The solution was allowed to warm slightly while stirring
for approximately 2.5 hours and then was re-cooled to 0.degree. C.
and stirred for 1.5 hours. Then approximately 20 mL of Rochelle's
salt was added carefully followed by ethyl acetate and the mixture
was stirred overnight. Then the layers were separated and the
aqueous layer was extracted two more times with ethyl acetate. The
combined organic layers were washed with brine, dried over
magnesium sulfate, concentrated and purified by chromatography
(silica gel, 20% ethyl acetate in hexanes) to afford a white solid.
The enantiomers were resolved by supercritical fluid chromatography
(Chiralpak AD, 10% methanol in supercritical carbon dioxide) and
the resulting solid was taken up with dichloromethane, dried over
magnesium sulfate and concentrated to afford
{3-(2,6-dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methanol
(1.35 g, 6%) .sup.1H-1-NMR (400 MHz, DMSO-d.sub.6) .delta.
7.61-7.51 (m, 3H), 4.85 (t, J=5 Hz, 1H), 4.20-4.06 (m, 2H),
3.15-3.10 (m, 1H), 1.69-1.62 (m, 2H), 1.29 (d, J=7 Hz, 3H), 0.79
(t, J=7 Hz, 3H).
14b) Methyl
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylate
##STR00146##
[0629] To a solution of methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (154 mg, 0.550 mmol),
triphenylphosphine (144 mg, 0.550 mmol) and
{3-(2,6-dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methanol
(150 mg, 0.500 mmol) in dichloromethane (1.5 mL) was added
diisopropyl azodicarboxylate (0.099 mL, 0.550 mmol) dropwise. The
solution was heated in a microwave reactor at 90.degree. C. for 10
minutes. The solution was adsorbed onto silica gel and purified by
chromatography (silica gel, 0-60% ethyl acetate in hexanes gradient
elution). A second purification was done by chromatography (silica
gel, 1% methanol in dichloromethane) to afford methyl
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylate (144 mg 0.256 mmol, 51%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (d, J=9 Hz, 1H),
8.28 (s, 1H), 8.18-8.10 (m, 3H), 7.73 (d, J=8 Hz, 2H), 7.63-7.51
(m, 3H), 6.92 (d, J=8 Hz, 2H), 4.88 (app q, J=12 Hz, 2H), 3.94 (s,
3H), 3.27-3.22 (m, 1H), 1.73-1.66 (m, 2H), 1.32 (d, J=7 Hz, 3H),
0.83 (t, J=7 Hz, 3H).
14c)
6-{4-[({3-(2,6-Dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}m-
ethyl)oxy]phenyl}-2-quinolinecarboxylic acid
##STR00147##
[0631] To a solution of methyl
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylate (50 mg, 0.089 mmol) in 2:1
tetrahydrofuran:methanol (1.5 mL) was added 1 N sodium hydroxide
(0.13 mL, 0.13 mmol). The solution was heated in a microwave
reactor at 80.degree. C. for 10 minutes and then 1 N hydrochloric
acid (0.13 mL, 0.13 mmol) was added. The solution was concentrated
and the residue was taken up with ethyl acetate and water. The
layers were separated and the organic layer was dried over
magnesium sulfate and concentrated to afford
6-{4-[({3-(2,6-dichlorophenyl)-5-[(1R)-1-methylpropyl]-4-isoxazolyl}methy-
l)oxy]phenyl}-2-quinolinecarboxylic acid (45 mg, 92%). .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 13.37 (br s, 1H), 8.53 (d, J=9 Hz,
1H), 8.27 (s, 1H), 8.18-8.08 (m, 3H), 7.73 (d, J=9 Hz, 2H),
7.63-7.51 (m, 3H), 6.99 (d, J=9 Hz, 2H), 4.88 (app q, J=12 Hz, 2H),
3.31-3.22 (m, 1H), 1.73-1.66 (m, 2H), 3.32 (d, J=7 Hz, 3H), 0.83
(t, J=7 Hz, 3H). HRMS (ESI) C.sub.30H.sub.24Cl.sub.2N.sub.2O.sub.4
calculated: 547.1186 (M+H).sup.+, found: 547.1186 (M+H).sup.+.
Example 15
6-[4-({3-[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propyl}oxy-
)phenyl]-2-quinolinecarboxylic acid
##STR00148##
[0632] 15a)
3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolecarbaldehyde
##STR00149##
[0634] To a solution of
[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methanol
(2.40 g, 8.39 mmol) in dichloromethane (160 mL) was added
pyridinium chlorochromate-silica gel (20.2 wt %, 44.7 g, 41.9
mmol). The mixture was stirred for approximately 1.5 hours and then
run through a silica gel pad and concentrated to afford
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolecarbaldehyde
(2.24 g, 7.88 mmol, 94%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.89 (s, 1H), 7.64-7.55 (m, 3H), 3.83 (septet, J=7 Hz, 1H),
1.39 (d, J=7 Hz, 6H).
15b)
3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-[(E/Z)-2-(methyloxy)etheny-
l]isoxazole
##STR00150##
[0636] To a solution of (methoxymethyl)triphenylphosphonium
chloride (8.09 g, 23.6 mmol) in tetrahydrofuran (10 mL) at
0.degree. C. was added potassium tert-butoxide (2.65 g, 23.6 mmol)
in tetrahydrofuran (40 mL) dropwise. The solution was stirred at
0.degree. C. for approximately 10 minutes and was transferred
dropwise to a stirring solution of
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolecarbaldehyde
(2.24 g, 7.88 mmol) in tetrahydrofuran (50 mL) at 0.degree. C. The
mixture was stirred at 0.degree. C. for approximately 45 minutes
and brine was added. The mixture was poured into ether and the
layers were separated. The organic layer was washed again with
brine, dried over magnesium sulfate, concentrated and purified by
chromatography (silica gel, 0-20% isopropanol in hexanes gradient
elution) to afford 3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-
RE/Z)-2-(methyloxy)ethenyl]isoxazole (2.24 g, 7.16 mmol, 91%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 7.66-7.47 (m, 3H), 6.15
(d, J=13 Hz, approximately 1H, major isomer), 6.12 (d, J=7 Hz,
approximately 1 H, minor isomer), 5.39 (d, J=13 Hz, approximately
1H, major isomer), 4.27 (d, J=7 Hz, approximately 1H, minor
isomer), 3.45 (s, 3H), 3.31 (septet, J=7 Hz, 1H), 1.28 (d, J=7 Hz,
6H).
15c)
3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-[(2E/Z)-3-(methyloxy)-2-pr-
open-1-yl]isoxazole
##STR00151##
[0638] To a solution of
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-[(E/Z)-2-(methyloxy)ethenyl]is-
oxazole (1.05 g, 3.35 mmol) in tetrahydrofuran (10 mL) was added 1
N hydrochloric acid (2.35 mL, 2.35 mmol). The mixture was heated in
a microwave reactor at 140.degree. C. for 10 minutes and was poured
into ether, washed twice with saturated sodium bicarbonate, then
with brine, and then was dried over magnesium sulfate, concentrated
and stored in a freezer overnight. To a separate solution of
(methoxymethyl)triphenylphosphonium chloride (4.05 g, 11.8 mmol) in
tetrahydrofuran (15 mL) at 0.degree. C. was added dropwise a
solution potassium tert-butoxide (1.32 g, 11.8 mmol) in
tetrahydrofuran cooled to 0.degree. C. The mixture was stirred at
0.degree. C. and then transferred to a solution of the above
aldehyde in tetrahydrofuran (15 mL). The solution was stirred for
approximately 15 minutes and brine was added. The mixture was
poured into ether and the layers were separated. The aqueous layer
was extracted with ether and the combined organic layers were dried
over magnesium sulfate, concentrated and purified by chromatography
(silica gel, 0-30% ethyl acetate in hexanes gradient elution) to
afford
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-[(2E/Z)-3-(methyloxy)-2-propen-
-1-yl]isoxazole (456 mg, 35%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. 7.62-7.51 (m, 3H), 5.99 (d, J=12 Hz, approximately 1H,
major isomer), 5.81 (d, J=7 Hz, approximately 1H, minor isomer),
4.47-4.41 (m, approximately 1H, major isomer), 4.09-4.03 (m,
approximately 1H, minor isomer), 3.30 (s, 3H), 3.28 (septet, J=7
Hz, 1H), 2.91-2.85 (m, 2H), 1.27 (d, J=7 Hz, 6H). LRMS (APCI) m/z
326 (M+H).sup.+.
15d)
3-[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propanal
##STR00152##
[0640] To a solution of
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-[(2E/Z)-3-(methyloxy)-2-propen-
-1-yl]isoxazole (241 mg, 0.739 mmol) in tetrahydrofuran (2.5 mL)
was added 1 N hydrochloric acid (0.517 mL, 0.517 mmol). The
solution was heated in a microwave reactor to 80.degree. C. for 10
minutes and was poured into ether, washed twice with saturated
sodium bicarbonate then with brine, and then was dried over
magnesium sulfate and concentrated to afford
3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propanal
(227 mg, 98%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 9.66 (s,
1H), 7.43-7.32 (m, 3H), 3.24 (septet, J=7 Hz, 1H), 2.59-2.44 (m,
4H), 1.38 (d, J=7 Hz, 6H).
15e)
3-[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]-1-propanol
##STR00153##
[0642] To a solution of
3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propanal
(220 mg, 0.705 mmol) in methanol (7 mL) at 0.degree. C. was added
sodium borohydride (67 mg, 1.8 mmol). The solution was stirred at
0.degree. C. for approximately 20 minutes and was then poured into
aqueous ammonium chloride. The mixture was extracted twice with
ether. The combined organic layers were dried over magnesium
sulfate and concentrated to afford
3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]-1-propan-
ol (204 mg, 92%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
7.42-7.28 (m, 3H), 3.53 (t, J=6 Hz, 2H), 3.21 (septet, J=7 Hz, 1H),
3.35 (t, J=8 Hz, 2H), 1.57-1.50 (m, 2H), 1.38 (d, J=7 Hz, 6H).
15f) Methyl
6-[4-({3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propyl}ox-
y)phenyl]-2-quinolinecarboxylate
##STR00154##
[0644] To a solution of methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (31 mg, 0.11 mmol),
3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]-1-propanol
(35 mg, 0.11 mmol) and triphenylphosphine (29 mg, 0.11 mmol) in
dichloromethane (1 mL) was added diisopropyl azodicarboxylate
(0.020 mL, 0.11 mmol). The solution was heated in a microwave
reactor at 90.degree. C. for 10 minutes and was then adsorbed onto
silica gel and purified by chromatography (silica gel, 0-35% ethyl
acetate in hexanes gradient elution) to afford methyl
6-[4-({3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propyl}ox-
y)phenyl]-2-quinolinecarboxylate (14 mg, 22%). .sup.1H-NMR (400
MHz, CDCl.sub.3) .delta. 8.33 (m, 2H), 8.21 (d, J=8 Hz, 1H),
8.03-7.99 (m, 2H), 7.63 (d, J=9 Hz, 2H), 7.40-7.28 (m, 3H), 6.91
(d, J=9 Hz, 2H), 4.09 (s, 3H), 3.89 (t, J=6 Hz, 2H), 3.19 (septet,
J=7 Hz, 1H), 2.51 (t, J=6 Hz, 2H), 1.82-1.76 (m, 2H), 1.33 (d, j=7
Hz, 6H).
15 g)
6-[4-({3-[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]prop-
yl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00155##
[0646] To a solution of methyl
6-[4-({3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propyl}ox-
y)phenyl]-2-quinolinecarboxylate (14 mg, 0.024 mmol) in 1:1
tetrahydrofuran:methanol (1 mL) was added 1 N sodium hydroxide
(0.089 mL, 0.089 mmol). The solution was heated in a microwave
reactor at 90.degree. C. for 10 minutes. Then 1 N hydrochloric acid
(0.089 mL, 0.089 mmol) was added and the mixture was concentrated.
Water and ethyl acetate were added to the residue and the layers
were separated. The ethyl acetate layer was dried over magnesium
sulfate and concentrated to afford 12 mg of
6-[4-({3-[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]propyl-
.delta.oxy)phenyl]-2-quinolinecarboxylic acid as a yellow solid
(0.021 mmol, 89%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 8.42
(d, J=9 Hz, 1H), 8.28 (d, J=9 Hz, 1H), 8.19 (d, J=9 Hz, 1H),
8.06-8.04 (m, 2H), 7.64 (d, J=9 Hz, 2H), 7.41-7.29 (m, 3H), 6.93
(d, J=9 Hz, 2H), 3.90 (t, J=6 Hz, 2H), 3.20 (septet, J=7 Hz, 1H),
2.52 (t, J=6 Hz, 2H), 1.83-1.77 (m, 2H), 1.34 (d, J=7 Hz, 6H). HRMS
(ESI) C.sub.31H.sub.26Cl.sub.2N.sub.2O.sub.4 calculated: 561.1342
(M+H).sup.+, found: 561.1344 (M+H).sup.+.
Example 16
7-[4-({[3-{[(2,6-Dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl-
]methyl}oxy)phenyl]-3-isoquinolinecarboxylic acid
##STR00156##
[0647] 16a) (1,1-Dimethylethyl)oxyethanal oxime
##STR00157##
[0649] To a stirring solution of ethylene glycol tent-butyl ether
(27.5 mL, 209 mmol) and triethylamine (87.5 mL, 628 mmol) in
dichloromethane (600 mL) at 0.degree. C. was added, over a period
of approximately 45 minutes, a solution of sulfur trioxide-pyridine
complex (100 g, 628 mmol) in dimethylsulfoxide (600 mL) that had
been stirring for approximately 25 minutes. The mixture was allowed
to warm to ambient temperature and stir over 6 hours and was then
poured into ether, washed three times with 10% aqueous citric acid,
then brine and was concentrated. The residue was taken up with
ethanol (2.65 L) and filtered into a stirring solution of
hydroxylamine hydrochloride (16.0 g, 230 mmol) and sodium hydroxide
(9.20 g, 230 mmol) in water (125 mL). The solution was heated to
approximately 90.degree. C. and stirred for approximately 17 hours.
The mixture was then concentrated and the residue was taken up with
ethyl acetate and washed twice with water containing sodium
chloride. The combined aqueous layers were back-extracted with
ethyl acetate and the combined organic layers were dried over
magnesium sulfate, concentrated and purified by chromatography
(silica gel, 15% ethyl acetate in hexanes) to afford
1,1-dimethylethyl)oxyethanal oxime (8.59 g, 31%). .sup.1H-NMR (400
MHz, DMSO-d.sub.6) .delta. 11.02 (s, 0.5H), 10.73 (s, 0.5H), 7.25
(t, J=6 Hz, 0.5H), 6.67 (t, J=4 Hz, 0.5H). 4.11 (d, J=4 Hz, 1H),
3.89 (d, J=6 Hz, 1H), 1.12 (s, 9H).
16b) Methyl
3-{[(1,1-dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyla-
te
##STR00158##
[0651] To a solution of (1,1-dimethylethyl)oxyethanal oxime (8.59
g, 65.5 mmol) in N,N-dimethylformamide (50 mL) was added
N-chlorosuccinimide (8.45 g, 65.5 mmol). The solution was stirred
for approximately 1 hour. The solution was poured into ether and
washed twice with water. The organic layer containing the crude
imidoyl chloride was then washed with brine, was dried over
magnesium sulfate and concentrated. Then to a solution of methyl
isobutyrylacetate (8.86 mL, 78.6 mmol) in tetrahydrofuran (40 mL)
at 0.degree. C. was added a 0.5 M solution of sodium methoxide in
methanol (157 mL, 78.6 mmol). After stirring for approximately 5
minutes the above imidoyl chloride was added in tetrahydrofuran (30
mL). A solid was observed to precipitate. After the addition was
complete the mixture was allowed to stir and warm to ambient
temperature overnight. Then the solution was poured into ether and
washed with water containing sodium chloride, dried over magnesium
sulfate and concentrated to afford methyl
3-{[(1,1-dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyla-
te (10.6 g, 63%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 4.52
(s, 2H), 3.77 (s, 3H), 3.67 (septet, J=7 Hz, 1H), 1.25 (d, J=7 Hz,
6H), 1.18 (s, 9H).
16c) Methyl
3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate
##STR00159##
[0653] To a solution of methyl
3-{[(1,1-dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyla-
te (500 mg, 1.96 mmol) in dichloromethane (97 mL) was added
trifluoroacetic acid (97 mL, 1.25 mol). The solution was stirred at
ambient temperature for approximately 45 minutes and then
concentrated. The residue was taken up with ethyl acetate and
poured into saturated sodium bicarbonate. Solid sodium bicarbonate
followed by solid sodium carbonate were added until the pH was
basic. The aqueous layer was extracted with ethyl acetate and the
combined organic layers were dried over magnesium sulfate,
concentrated and purified by chromatography (silica gel, 25% ethyl
acetate in hexanes) to afford the methyl
3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate (304 mg,
78%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 5.29 (t, J=6 Hz,
1H), 4.62 (d, J=6 Hz, 2H), 3.77 (s, 3H), 3.68 (septet, J=7 Hz, 1H),
1H), 1.25 (d, J=7 Hz, 6H).
16d) Methyl
3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyl-
ate
##STR00160##
[0655] To a solution of 2,6-dimethylphenol (184 mg, 1.51 mmol),
triphenylphosphine (396 mg, 1.51 mmol), and methyl
3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate (300 mg,
1.51 mmol) in toluene (4 mL) was slowly added diisopropyl
azodicarboxylate (0.272 mL, 1.51 mmol). The solution was heated in
a microwave reactor at 90.degree. C. for 10 minutes and then
allowed to stand at ambient temperature overnight. The solution was
concentrated and purified by chromatography (silica gel, 2.5%
acetone in hexanes) to afford methyl
3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyl-
ate (329 mg, 72%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
7.02-6.90 (m, 3H), 5.00 (s, 2H), 3.74 (s, 3H), 3.72 (septet, J=7
Hz, 1H), 2.17 (s, 6H), 1.28 (d, J=7 Hz, 6H).
16e)
[3-{[(2,6-Dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl]m-
ethanol
##STR00161##
[0657] To a solution of methyl
3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyl-
ate (183 mg, 0.603 mmol) in tetrahydrofuran (2 mL) at 0.degree. C.
was added a 1.5 M solution of diisobutylaluminum hydride in toluene
(1.33 mL, 1.99 mmol) dropwise. The solution was allowed to warm
slowly to ambient temperature and after approximately 5 hours was
re-cooled to 0.degree. C. Then 10 mL of Rochelle's salt was slowly
added followed by ethyl acetate. The mixture was allowed to stir
overnight. The layers were separated and the aqueous layer was
extracted with ethyl acetate. The combined organic layers were
washed with brine, dried over magnesium sulfate and concentrated to
afford
[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl]metha-
nol (158 mg, 95%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
7.03-6.91 (m, 3H), 4.98 (t, J=5 Hz, 1H), 4.83 (s, 2H), 4.39 (d, J=5
Hz, 2H), 3.29 (septet, J=7 Hz, 1H), 2.19 (s, 6H), 1.24 (d, J=7 Hz,
6H).
16f) Methyl
7-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-3-isoquinolinecarboxylate
##STR00162##
[0659] To a solution of triphenylphosphine (160 mg, 0.611 mmol),
[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl]metha-
nol (153 mg, 0.556 mmol) and methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate (155 mg, 0.556 mmol)
in dichloromethane (1.5 mL) was added diisopropyl azodicarboxylate
(0.110 mL, 0.611 mmol) dropwise. The solution was heated in a
microwave reactor at 90.degree. C. for 10 minutes and then purified
by chromatography (silica gel, 0-25% acetone in hexanes gradient
elution) to afford methyl
7-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-3-isoquinolinecarboxylate (109 mg, 37%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 9.11 (s, 1H), 8.64 (s,
1H), 8.50 (s, 1H), 8.29-8.20 (m, 2H), 7.86 (d, J=9 Hz, 2H), 7.19
(d, J=9 Hz, 2H), 7.00-6.90 (m, 3H), 5.09 (s, 2H), 4.90 (s, 2H),
3.91 (s, 3H), 3.41 (septet, J=7 Hz, 1H), 2.14 (s, 6H), 1.28 (d, J=7
Hz, 6H).
16 g)
7-[4-({[3-{[(2,6-Dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-3-isoquinolinecarboxylic acid
##STR00163##
[0661] To a solution of methyl
7-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-3-isoquinolinecarboxylate (44 mg, 0.082 mmol)
in 2:1 tetrahydrofuran:methanol (3 mL) was added 0.123 mL of 1 N
sodium hydroxide. The solution was heated in a microwave reactor at
100.degree. C. for 500 seconds. Then 1N hydrochloric acid (0.123
mL, 0.123 mmol) was added and the solution was concentrated. The
residue was taken up with ethyl acetate and water. The aqueous
layer was extracted with ethyl acetate and the combine organic
layers were washed with brine, dried over magnesium sulfate and
concentrated. The resulting solid was dried under vacuum overnight
and then suspended in ethyl acetate and washed three times with
water. The solvent was evaporated to afford
7-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-3-isoquinolinecarboxylic acid (32 mg, 75%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 9.41 (s, 1H), 8.61 (s,
1H), 8.50 (s, 1H), 8.24-8.19 (m, 2H), 7.86 (d, J=9 Hz, 2H), 7.19
(d, J=9 Hz, 2H), 7.00-6.91 (m, 3H), 5.09 (s, 2H), 4.90 (s, 2H),
3.41 (septet, J=7 Hz, 1H), 2.14 (s, 6H), 1.27 (d, J=7 Hz, 6H). HRMS
(ESI) C.sub.32H.sub.30N.sub.2O.sub.5 calculated: 523.2228
(M+H).sup.+, found: 523.2230 (M+H).sup.+.
Example 17
7-(4-{[(5-Cyclobutyl
-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)methyl]oxy]phenyl)-3-is-
oquinolinecarboxylic acid
##STR00164##
[0662] 17a) Ethyl 5-cyclobutyl
-3-{[(1,1-dimethylethyl)oxy]methyl}-4-isoxazolecarboxylate
##STR00165##
[0664] To a solution of (1,1-dimethylethyl)oxyethanal oxime (1.71
g, 13.0 mmol) in N,N-dimethylformamide (10 mL) was added
N-chlorosuccinimide (1.74 g, 13.0 mmol) while the flask was in a
water bath. After approximately 10 minutes the flask was removed
from the water bath and the solution was stirred for approximately
1 hour. The solution, containing the crude imidoyl chloride, was
then poured into ether and washed twice with water followed by
brine, and then was dried over magnesium sulfate and concentrated.
Then to a separate solution of ethyl 3-cyclobutyl-3-oxopropanoate
(2.66 g, 15.6 mmol) in tetrahydrofuran (8 mL) at 0.degree. C. was
added sodium ethoxide (21 wt % solution in ethanol, 5.82 mL, 15.6
mmol). The above imidoyl chloride was diluted with tetrahydrofuran
(6 mL) then added to the solution containing the ketoester. A solid
was seen to precipitate. After the addition the solution was
allowed to warm to ambient temperature and stirred overnight. The
mixture was then poured into ether and washed twice with water
followed by brine, dried over magnesium sulfate and concentrated to
afford ethyl
5-cyclobutyl-3-{[(1,1-dimethylethyl)oxy]methyl}-4-isoxazolecarboxylate
(3.13 g, 87%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 4.53 (s,
2H), 4.22 (q, J=7 Hz, 2H), 4.17-4.05 (m, 1H), 2.33-2.27 (m, 4H),
2.09-2.03 (m, 1H), 1.95-1.86 (m, 1H), 1.26 (t, J=7 Hz, 3H), 1.17
(s, 9H).
17b) Ethyl
5-cyclobutyl-3-(hydroxymethyl)-4-isoxazolecarboxylate
##STR00166##
[0666] To a solution of ethyl
5-cyclobutyl-3-{[(1,1-dimethylethyl)oxy]methyl}-4-isoxazole
carboxylate (1.40 g, 5.48 mmol) in dichloromethane (250 mL) was
added trifluoroacetic acid (250 mL, 3.25 mol). The solution was
stirred for approximately 30 minutes and then concentrated. The
residue was taken up with ethyl acetate and poured into water. The
mixture was stirred as solid sodium carbonate was added until the
pH was basic. The layers were separated and the aqueous layer was
extracted with ethyl acetate. The combined organic layers were
washed with brine, dried over magnesium sulfate and concentrated.
The residue was purified by chromatography (silica gel, 20% ethyl
acetate in hexanes) to afford ethyl
5-cyclobutyl-3-(hydroxymethyl)-4-isoxazolecarboxylate (848 mg,
69%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 5.27 (t, J=6 Hz,
1H), 4.63 (d, J=6 Hz, 2H), 4.21 (q, J=7 Hz, 2H), 4.18-4.08 (m, 1H),
2.33-2.27 (m, 4H), 2.10-2.00 (m, 1H), 1.90-1.83 (m, 1H), 1.26 (t,
J=7 Hz, 3H).
17c) Ethyl
5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolecar-
boxylate
##STR00167##
[0668] To a solution of 2,6-dimethylphenol (503 mg, 4.12 mmol),
triphenylphosphine (1.08 g, 4.12 mmol) and ethyl
5-cyclobutyl-3-(hydroxymethyl)-4-isoxazolecarboxylate (929 mg, 4.12
mmol, from multiple batches) in toluene (10 mL) was added
diisopropyl azodicarboxylate (0.741 mL, 4.12 mmol) dropwise. The
solution was heated in a microwave reactor to 90.degree. C. for 10
minutes. The solution was then concentrated and the residue was
purified by chromatography (silica gel, 2.5% acetone in hexanes) to
afford ethyl
5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolecarboxylate
(1.11 g, 81%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.7.01-6.90
(m, 3H), 5.02 (s, 2H), 4.20 (q, J=7 Hz, 2H), 4.20-4.10 (m, 1H),
2.37-2.30 (m, 4H), 2.14 (s, 6H), 2.11-2.01 (m, 1H), 1.95-1.85 (m,
1H), 1.22 (t, J=7 Hz, 3H).
17d)
(5-Cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)methan-
ol
##STR00168##
[0670] To a solution of ethyl
5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolecarboxylate
(1.08 g, 3.26 mmol) at 0.degree. C. was added a 1.5 M solution of
diisobutylaluminum hydride in toluene (7.18 mL, 10.8 mmol) at a
slow pace. The solution was allowed to stir while warming slightly
for approximately 1.5 hours and then was re-cooled to 0.degree. C.
Then approximately 40 mL of Rochelle's salt was slowly added
followed by ethyl acetate. The mixture was allowed to warm to
ambient temperature and stirred overnight. Then the layers were
separated and the aqueous layer was extracted with ethyl acetate.
The combined organic layers were washed with brine, dried over
magnesium sulfate and concentrated to afford
(5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)methanol
(937 mg, 100%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
7.03-6.91 (m, 3H), 4.97 (t, J=5 Hz, 1H), 4.83 (s, 2H), 4.35 (d, J=5
Hz, 2H), 3.85-3.77 (m, 1H), 2.362.26 (m, 4H), 2.14 (s, 6H),
2.10-1.98 (m, 1H), 1.94-1.84 (m, 1H).
17e) Methyl
7-(4-{[(5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)met-
hyl]oxy}phenyl)-3-isoquinolinecarboxylate
##STR00169##
[0672] To a solution of methyl
7-(4-hydroxyphenyl)-3-isoquinolinecarboxylate (90 mg, 0.32 mmol),
triphenylphosphine (93 mg, 0.35 mmol), and
(5-cyclobutyl-3-{[(2,6-dimethyl
phenyl)oxy]methyl}-4-isoxazolyl)methanol (93 mg, 0.32 mmol) in
dichloromethane (1.5 mL) was added diisopropyl azodicarboxylate
(0.064 mL, 0.35 mmol) dropwise. The solution was heated in a
microwave reactor at 90.degree. C. for 10 minutes. The solution was
concentrated, adsorbed onto silica gel and purified by
chromatography (silica gel, 0-25% acetone in hexanes gradient
elution) to afford methyl
7-(4-{[(5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)met-
hyl]oxy}phenyl)-3-isoquinolinecarboxylate (103 mg, 58%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 9.41 (s, 1H), 8.64 (s,
1H), 8.50 (s, 1H), 8.26-8.19 (m, 2H), 7.85 (d, J=9 Hz, 2H), 7.16
(d, J=9 Hz, 2H), 7.00-6.90 (m, 3H), 5.06 (s, 2H), 4.90 (s, 2H),
3.99-3.90 (m, 1H), 3.91 (s, 3H), 2.37-2.30 (m, 4H), 2.14 (s, 6H),
2.09-2.01 (m, 1H), 1.98-1.89 (m, 1H).
17f)
7-(4-{[(5-Cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl-
)methyl]oxy}phenyl)-3-isoquinolinecarboxylic acid
##STR00170##
[0674] To a solution of methyl
7-(4-{[(5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazolyl)met-
hyl]oxy}phenyl)-3-isoquinolinecarboxylate (50 mg, 0.091 mmol) in
2:1 tetrahydrofuran:methanol (3 mL) was added 1 N sodium hydroxide
(0.14 mL, 0.14 mmol). The solution was heated in a microwave
reactor at 100.degree. C. for 500 seconds. Then 1 N hydrochloric
acid (0.14 mL, 0.14 mmol) was added and the mixture was
concentrated. The residue was taken up with ethyl acetate and
water. The layers were separated and the organic layer was washed
with water, dried over magnesium sulfate and concentrated to afford
7-(4-{[(5-cyclobutyl-3-{[(2,6-dimethylphenyl)oxy]methyl}-4-isoxazo-
lyl)methyl]oxy}phenyl)-3-isoquinolinecarboxylic acid (41 mg, 84%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 9.41 (s, 1H), 8.61 (s,
1H), 8.49 (s, 1H), 8.24-8.18 (m, 2H), 7.85 (d, J=9 Hz, 2H), 7.16
(d, J=9 Hz, 2H), 7.00-6.90 (m, 3H), 5.06 (s, 2H), 4.90 (s, 2H),
4.01-3.90 (m, 1H), 2.39-2.30 (m, 4H), 2.14 (s, 6H), 2.10-2.00 (m,
1H), 1.90-1.89 (m, 1H),. HRMS (ESI) C.sub.33H.sub.30N.sub.2O.sub.5
calculated: 535.2228 (M+H).sup.+, found: 535.2228 (M+H).sup.+.
Example 18
6-[4-({[3-{[(2,6-Dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl-
]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00171##
[0675] 18a)
[3-{[(1,1-Dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl]methan-
ol
##STR00172##
[0677] To a solution of methyl
3-{[(1,1-dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyla-
te (21.5 g, 84.2 mmol) in tetrahydrofuran (250 mL) at 0.degree. C.
was added a 1.5 M solution of diisobutylaluminum hydride in toluene
(185 mL, 278 mmol) slowly. The solution was allowed to warm slowly
to ambient temperature overnight then was re-cooled to 0.degree. C.
and approximately 250 mL of Rochelle's salt was added dropwise
followed by approximately 300 mL of ethyl acetate. An additional
250 mL of Rochelle's salt and 500 mL of ethyl acetate were added
and the mixture was stirred at 0.degree. C. for approximately 20
minutes and then at ambient temperature for approximately 4 hours.
The mixture was filtered. The layers were separated and the aqueous
layer was extracted with ethyl acetate. The combined organic layers
were dried over magnesium sulfate, concentrated, and purified by
chromatography (silica gel, 20% ethyl acetate in hexanes) to afford
[3-{[(1,1-dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl]methan-
ol (15.2 g, 91%). .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 4.76
(t, J=5 Hz, 1H), 4.39 (s, 2H), 4.32 (d, J=5 Hz, 2H), 3.24 (septet,
J=7 Hz, 1H), 1.21 (d, J=7 Hz, 6H), 1.18 (s, 9H).
18b) Methyl
6-[4-({[3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylate
##STR00173##
[0679] To a solution of
[3-{[(1,1-dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl]methan-
ol (407 mg, 1.79 mmol), methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (500 mg, 1.79 mmol) and
triphenylphosphine (517 mg, 1.97 mmol) in dichloromethane (4 mL)
was added diisopropyl azodicarboxylate (0.354 mL, 1.97 mmol)
dropwise. The solution was heated in a microwave reactor at
100.degree. C. for 10 minutes and then adsorbed onto silica gel and
purified by chromatography (silica gel, 0-40% ethyl acetate in
hexanes gradient elution) to afford a solid which was dissolved in
dichloromethane (114 mL). Trifluoroacetic acid (114 mL, 1.48 mol)
was added at a rapid pace. The mixture was stirred for
approximately 1.5 hours and then concentrated. The residue was
taken up with ethyl acetate and poured into water. Then solid
sodium carbonate was added until the pH was between 9 and 10. The
layers were separated and the ethyl acetate layer was dried over
magnesium sulfate and concentrated. The resulting solid was washed
with ether to afford methyl
6-[4-({[3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylate (545 mg, 54%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.57 (d, J=9 Hz, 1H), 8.33 (s, 1H), 8.18 (s,
2H), 8.12 (d, J=9 Hz, 1H), 7.83 (d, J=9 Hz, 2H), 7.17 (d, J=9 Hz,
2H), 5.46 (br s, 1H), 5.06 (s, 2H), 4.54 (s, 2H), 3.94 (s, 3H),
3.33 (septet, J=7 Hz, 1H), 1.24 (d, J=7 Hz, 6H). LRMS (ESI) m/z 433
(M+H).sup.+.
18c) Methyl
6-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylate
##STR00174##
[0681] To a solution of methyl
6-[4-({[3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylate (60 mg, 0.14 mmol), 2,6-dimethylphenol
(17 mg, 0.14 mmol) and triphenylphosphine (40 mg, 0.15 mmol) in
dichloromethane (1.5 mL) was added diisopropyl azodicarboxylate
(0.028 mL, 0.15 mmol) dropwise. The solution was stirred at ambient
temperature for approximately 1.5 hours and then in a microwave
reactor at 100.degree. C. for 10 minutes. The solution was adsorbed
onto silica gel and purified by chromatography (silica gel, 0-40%
ethyl acetate gradient elution). The resulting residue was taken up
with ether and sonicated until a solid precipitated. The solid was
isolated by filtration to afford methyl
6-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylate (23 mg, 31%).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.37-8.32 (m, 2H),
8.23-8.21 (m, 1H), 8.05-8.01 (m, 2H), 7.69 (d, J=9 Hz, 2H), 7.08
(d, J=9 Hz, 2H), 7.01-6.01 (m, 3H), 5.04 (s, 2H), 4.96 (s, 2H),
4.09 (s, 3H), 3.29 (septet, J=7 Hz, 1H), 2.23 (s, 6H), 1.38 (d, J=7
Hz, 6H).
18d)
6-[4-({[3-{[(2,6-Dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxa-
zolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00175##
[0683] To a solution of methyl
6-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylate (23 mg, 0.043 mmol) in
2:1 tetrahydrofuran:methanol (0.75 mL) was added 1 N sodium
hydroxide (0.064 mL, 0.064 mmol). The solution was heated in a
microwave reactor at 120.degree. C. for 500 seconds. The mixture
was concentrated and water was added followed by 1 N hydrochloric
acid (0.064 mL, 0.064 mmol). The mixture was extracted with ethyl
acetate and the combined organic layers were washed with brine and
concentrated to afford of
6-[4-({[3-{[(2,6-dimethylphenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylic acid as a yellow solid
(14 mg, 62%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.43 (d,
J=9 Hz, 1H), 8.29 (d, J=8 Hz, 1H), 8.21 (d, J=9 Hz, 1H), 8.07 (m,
2H), 7.70 (d, J=9 Hz, 2H), 7.10 (d, J=9 Hz, 2H), 7.01-6.62 (m, 3H),
5.06 (s, 2H), 4.97 (s, 2H), 3.30 (septet, J=7 Hz, 1H), 2.23 (s,
6H), 1.39 (d, J=7 Hz, 6H). HRMS (ESI)
C.sub.32H.sub.30N.sub.2O.sub.5 calculated: 437.2263 (M+H).sup.+,
found: 437.2263 (M+H).sup.+.
Example 19:
6-(4-{[(5-(1-Methylethyl)-3-{[(2,4,6-trifluorophenyl)oxy]methyl}-4-isoxazo-
lyl)methyl]oxy}phenyl)-2-quinolinecarboxylic acid
##STR00176##
[0684] 19a)
6-(4-{[(5-(1-Methylethyl)-3-{[(2,4,6-trifluorophenyl)oxy]methyl}-4-isoxaz-
olyl)methyl]oxy}phenyl)-2-quinolinecarboxylic acid
[0685] To a solution of 2,4,6-trifluorophenol (18 mg, 0.12 mmol),
methyl
6-[4-({[3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinoline carboxylate (52 mg, 0.12 mmol) and
triphenylphosphine (35 mg, 0.13 mmol) in dichloromethane (2 mL) was
added diisopropyl azodicarboxylate (0.024 mL, 0.13 mmol) slowly.
The solution was heated in a microwave reactor at 100.degree. C.
for 10 minutes. The solution was adsorbed onto silica gel and
purification was done by chromatography (0-70% ethyl acetate in
hexanes) to afford a solid which was dissolved in 2:1
tetrahydrofuran:methanol (1.5 mL). Then 1 N sodium hydroxide (0.19
mmol, 0.19 mmol) was added. The solution was subjected to microwave
radiation at 120.degree. C. for 500 seconds. The solution was
concentrated then diluted with water before the addition of 1 N
hydrochloric acid (0.19 mmol, 0.19 mmol). The resulting solid was
extracted with ethyl acetate twice and the combined organic layers
were dried over magnesium sulfate and concentrated. The resulting
solid was washed with methanol and dried to afford
6-(4-{[(5-(1-methylethyl)-3-{
[(2,4,6-trifluorophenyl)oxy]methyl}-4-isoxazolyl)methyl]oxy}phenyl)-2-qui-
nolinecarboxylic acid (24 mg, 35%). .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.44-8.06 (m, 5H), 7.69 (d, J=9 Hz, 2H), 7.11
(d, J=9 Hz, 2H), 6.65 (t, J=9 Hz, 2H), 5.23 (s, 2H), 5.12 (s, 2H),
3.29 (septet, J=7 Hz, 1H), 1.37 (d, J=7 Hz, 6H). HRMS (ESI)
C.sub.30H.sub.23F.sub.3N.sub.2O.sub.5 calculated: 549.1632
(M+H).sup.+, found: 549.1631(M+H).sup.+.
Example 20
6-[4-({[3-{[(2,6-Dichlorophenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolyl-
]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00177##
[0686] 20a)
6-[4-({[3-{[(2,6-Dichlorophenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
[0687] To a solution of triphenylphosphine (34 mg, 0.13 mmol),
2,6-dichlorophenol (21 mg, 0.13 mmol) and methyl
6-[4-({[3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylate (56 mg, 0.13 mmol) was added diisopropyl
azodicarboxylate (0.023 mL, 0.13 mmol). The solution was heated in
a microwave reactor at 100.degree. C. for 10 minutes. This heating
was repeated again for 10 minutes. The mixture was adsorbed onto
silica gel and purified by chromatography (silica gel, 0-60% ethyl
acetate in hexanes gradient elution) to afford a solid that was
dissolved in 2:1 tetrahydrofuran/methanol (1.5 mL). Then 1 N sodium
hydroxide (0.13 mL, 0.13 mmol) was added. The solution was heated
in a microwave reactor at 120.degree. C. for 500 seconds. The
mixture was concentrated and the residue was taken up with
methanol. Then 10% aqueous citric acid was added and the mixture
was extracted with ethyl acetate. The aqueous layer was acidified
with additional citric acid until the pH was approximately 2-3 and
was extracted one more time with ethyl acetate. The combined
organic layers were concentrated and the residue was taken up with
ether which was removed under vacuum. The resulting solid was
dissolved in ethyl acetate and the solution was washed with water
and brine and then concentrated to afford
6-[4-({[3-{[(2,6-dichlorophenyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylic acid (12 mg, 24%).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.36 (d, J=9 Hz, 1H),
8.24-8.19 (m, 2H), 8.02 (d, J=10 Hz, 2H), 7.66 (d, J=9 Hz, 2H),
7.24 (d, J=9 Hz, 2H), 7.08 (d, J=9 Hz, 2H), 6.99-6.97 (m, 1H) 5.15
(s, 4H), 3.27 (3.27 (septet, J=7 Hz, 1H), 1.34 (d, J=7 Hz, 6H).
HRMS (ESI) C.sub.30H.sub.24Cl.sub.2N.sub.2O.sub.5 calculated:
563.1135 (M+H).sup.+, found: 563.1130 (M+H).sup.+.
Example 21
6-[4-({[3-{[(2,6-Dichlorophenyl)amino]methyl}-5-(1-methylethyl)-4-isoxazol-
yl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00178##
[0688] 21a) N-(2,6-Dichlorophenyl)-2,2,2-trifluoroacetamide
##STR00179##
[0690] To a solution of 2,6-dichloroaniline (243 mg, 1.50 mmol) in
dichloromethane (50 mL) at 0.degree. C. was added trifluoroacetic
anhydride (0.254 mL, 1.80 mmol) dropwise. The solution was stirred
while the flask was in the cold bath for approximately 3 hours and
then at ambient temperature for approximately 1 hour. The solution
was concentrated and the residue was diluted with ethyl acetate,
washed with aqueous sodium bicarbonate, dried over magnesium
sulfate and concentrated to afford
N-(2,6-dichlorophenyl)-2,2,2-trifluoroacetamide (350 mg, 90%).
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 11.58 (s, 1H),
7.63-7.43 (m, 3H).
21b) Methyl
6-[4-({[3-{[(2,6-dichlorophenyl)(trifluoroacetyl)amino]methyl}-5-(1-methy-
lethyl)-4-isoxazolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate
##STR00180##
[0692] To a solution of methyl
6-[4-({[3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)pheny-
l]-2-quinolinecarboxylate (25 mg, 0.058 mmol),
N-(2,6-dichlorophenyl)-2,2,2-trifluoroacetamide (15 mg, 0.058 mmol)
and triphenylphosphine (30 mg, 0.12 mmol) in dichloromethane was
added di-tent-butyl azodicarboxylate (27 mg, 0.12 mmol). The
solution was stirred at ambient temperature for approximately 1.5
hours and then was adsorbed onto silica gel and purified by
chromatography (silica gel, 0-100% ethyl acetate in hexanes
gradient elution). The resulting mixture was further purified by
chromatography (silica gel, 2% methanol in dichloromethane) to
afford methyl
6-[4-({[3-{[(2,6-dichlorophenyl)(trifluoroacetyl)amino]methyl}-5-(1-methy-
lethyl)-4-isoxazolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate (10
mg, 26%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.36-8.33 (m,
2H), 8.22 (d, J=9 Hz, 1H), 8.06-8.01 (m, 2H), 7.71 (d, J=9 Hz, 2H),
7.42-7.29 (m, 3H), 7.11 (d, J=9 Hz, 2H), 5.05 (s, 2H), 5.04 (s,
2H), 4.05 (s, 3H), 3.27 (septet, J=7 Hz, 1H), 1.34 (d, J=7 Hz, 6H).
LRMS (APCI) m/z 672 (M+H).sup.-.
21c)
6-[4-({[3-{[(2,6-Dichlorophenyl)amino]methyl}-5-(1-methylethyl)-4-iso-
xazolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00181##
[0694] To a solution of methyl
6-[4-({[3-{[(2,6-dichlorophenyl)(trifluoroacetyl)amino]methyl}-5-(1-methy-
lethyl)-4-isoxazolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate (10
mg, 0.015 mmol) in 1:1 tetrahydrofuran:methanol (0.75 mL) was added
1 N sodium hydroxide (0.37 mL, 0.37 mmol). The solution was heated
in a microwave reactor at 90.degree. C. for 10 minutes. The
solution was concentrated and the residue was taken up with ethyl
acetate. Then 1 N hydrochloric acid (0.37 mL, 0.37 mmol) was added
followed by water. The layers were separated and the aqueous layer
was extracted one more time with ethyl acetate. The combined
organic layers were dried over magnesium sulfate and concentrated
to afford
6-[4-({[3-{[(2,6-dichlorophenyl)amino]methyl}-5-(1-methylethyl)-4-isoxazo-
lyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid (8 mg, 100%).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.46 (d, J=9 Hz, 1H),
8.31-8.24 (m, 2H), 8.11-8.07 (m, 2H), 7.69 (d, J=9 Hz, 2H),
7.25-7.20 (m, 2H), 7.07 (d, J=9 Hz, 2H), 6.84-6.80 (m, 1H), 4.96
(s, 2H), 4.60 (s, 2H), 3.24 (septet, J=7 Hz, 1H), 1.36 (d, J=7 Hz,
6H). HRMS (ESI) C.sub.30H.sub.25Cl.sub.2N.sub.3O.sub.4 calculated:
562.1300 (M+H).sup.+, found: 562.1292 (M+H).sup.+.
Example 22
6-[4-({[3-{[(2,6-Dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazoly-
l]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00182##
[0695] 22a) Methyl
3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazolecarboxy-
late
##STR00183##
[0697] To a solution of 2,6-dichlorobenzenethiol (530 mg, 2.96
mmol), methyl
3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate (629 mg,
2.46 mmol) and triphenylphosphine (1.29 g, 4.93 mmol) in
dichloromethane (20 mL) was added di-tert-butyl azodicarboxylate
(1.14 g, 4.93 mmol). The solution was stirred at ambient
temperature overnight. The solution was then adsorbed onto silica
gel and purified by chromatography (silica gel, 0-100% ethyl
acetate in hexanes gradient elution) to afford methyl
3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazolecarboxy-
late (886 mg, 100%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
7.52-7.35 (m, 3H), 4.18 (s, 3H), 3.72 (s, 2H), 3.64 (septet, J=7
Hz, 1H), 1.20 (d, J=7 Hz, 6H). LRMS (APCI) m/z 360 (M+H).sup.-.
22b)
[3-{[(2,6-Dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazolyl]-
methanol
##STR00184##
[0699] To a solution of methyl
3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazolecarboxy-
late (50 mg, 0.14 mmol) in tetrahydrofuran (0.5 mL) at 0.degree. C.
was slowly added a 1.5 M solution of diisobutylaluminum hydride in
toluene (0.46 mL, 0.69 mmol). The solution was allowed to warm
slowly to ambient temperature and stir overnight. The next day the
solution was cooled to 0.degree. C. and methanol (approximately
0.25 mL) was added followed by aqueous Rochelle's salt
(approximately 3 mL). The mixture was diluted with ethyl acetate
and allowed to warm to ambient temperature while stirring. The
layers were separated and the organic layer was dried over
magnesium sulfate and concentrated to afford
[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazolyl]meth-
anol (43 mg, 93%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
7.35-7.16 (m, 3H), 4.60 (s, 2H), 4.10 (s, 2H), 3.17 (septet, J=8
Hz, 1H), 1.28 (d, J=6 Hz, 6H).
22c) Ethyl 6-(4-hydroxyphenyl)-2-quinolinecarboxylate
##STR00185##
[0701] Palladium(II) acetate (17.1 mg, 76.0 .mu.mol) was added to
ethyl 6-bromo-2-quinolinecarboxylate (426.0 mg, 1.52 mmol, from
Example 32c), 4-hydroxy-phenyl-boronic acid (314.6 mg, 2.28 mmol),
triphenylphosphine (39.9 mg, 152.1 .mu.mol), and potassium
phosphate (1.13 g, 5.32 mmol). Then, dioxane (7.6 mL) was added to
the mixture, followed by water (152 .mu.L), and the reaction
mixture was heated open to the atmosphere at 60.degree. C. in an
oil bath for 16 hours, then allowed to cool to room temperature.
Water was added followed by ethyl acetate and the mixture was
filtered through Celite.RTM.. The filtrate was extracted with ethyl
acetate and the organic layer was dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 2:3 ethyl acetate:hexanes
to give 260.5 mg (58%) of ethyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 9.74 (s, 1H), 8.55 (d, J=9 Hz,
1H), 8.28 (s, 1H), 8.19-8.10 (m, 3H), 7.72 (d, J=9 Hz, 2H), 6.92
(d, J=9 Hz, 2H), 4.42 (q, J=7 Hz, 2H), 1.38 (t, J=7 Hz, 3H).
ESI-LCMS m/z 294 (M+H).sup.+.
22d) Ethyl
6-[4-({[3-{[(2,6-Dichlorophenyl)thio]methyl}-5-(1-methylethyl)--
4-isoxazolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate
##STR00186##
[0703] To a solution of
[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazolyl]meth-
anol (22 mg, 0.066 mmol), ethyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (19 mg, 0.066 mmol) and
triphenylphosphine (35 mg, 0.13 mmol) in dichloromethane (1 mL) was
added di-tent-butyl azodicarboxylate (30 mg, 0.13 mmol). The
solution was stirred at ambient temperature approximately 4.5 hours
and then was adsorbed onto silica gel. Purification was done by
chromatography (silica gel, 0-75% ethyl acetate in hexanes gradient
elution) to afford ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazol-
yl]methyl}oxy)phenyl]-2-quinolinecarboxylate (26 mg, 65%).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.36-8.31 (m, 2H), 8.20
(d, J=9 Hz, 1H), 8.03-8.00 (m, 2H), 7.69 (d, J=9 Hz, 2H), 7.32-7.13
(m, 3H), 7.08 (d, J=9 Hz, 2H), 5.08 (s, 2H), 4.56 (q, J=7 Hz, 2H),
4.16 (s, 2H), 3.20 (sept, J=7 Hz, 1H), 1.49 (t, J=7 Hz, 3H), 1.31
(d, J=7 Hz, 6H). LRMS (APCI) m/z 607 (M+H).sup.-.
22e)
6-[4-({[3-{[(2,6-Dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00187##
[0705] To a solution of ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazol-
yl]methyl}oxy)phenyl]-2-quinolinecarboxylate (5 mg, 0.008 mmol) in
1:1 tetrahydrofuran:methanol (0.5 mL) was added 1 N sodium
hydroxide (0.04 mL, 0.04 mmol). The solution was heated in a
microwave reactor at 90.degree. C. for 10 minutes and 1 N
hydrochloric acid (0.04 mL, 0.04 mmol) was added. The mixture was
concentrated and the residue was taken up with ethyl acetate and
washed with water. The organic layer was dried over magnesium
sulfate and concentrated to afford
6-[4-({[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazol-
yl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid (5 mg, 100%).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.44 (d, J=9 Hz, 1H),
8.29 (d, J=9 Hz, 1H), 8.21 (d, J=9 Hz, 1H), 8.10-8.07 (m, 2H), 7.70
(d, J=9 Hz, 2H), 7.34-7.14 (m, 3H), 7.10 (d, J=9 Hz, 2H), 5.08 (s,
2H), 4.16 (s, 2H), 3.20 (septet, J=7 Hz, 1H), 1.32 (d, J=7 Hz, 6H).
HRMS (ESI) C.sub.30H.sub.24Cl.sub.2N.sub.2O.sub.4S calculated:
579.0912 (M+H).sup.+, found: 579.0925 (M+H).sup.+.
Example 23
6-[4-({[3-{[(2,6-Dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl)-4-isoxa-
zolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00188##
[0706] 23a) Ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate
##STR00189##
[0708] To a solution of ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazol-
yl]methyl}oxy)phenyl]-2-quinolinecarboxylate (20 mg, 0.033 mmol) in
dichloromethane (1 mL) at 0.degree. C. was added
meta-chloroperoxybenzoic acid (15 mg, 0.066 mmol). The mixture was
stirred at 0.degree. C. for approximately 30 minutes. Aqueous
sodium bicarbonate was added and the mixture was extracted twice
with dichloromethane. The combined organic layers were dried over
magnesium sulfate, adsorbed onto silica gel and purified by
chromatography (silica gel, 0-100% ethyl acetate in hexanes
gradient elution) to afford ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate (12 mg, 0.019
mmol, 57%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.39 (d, J=9
Hz, 1H), 8.23 (d, J=9 Hz, 2H), 8.05-8.03 (m, 2H), 7.68 (d, J=9 Hz,
2H), 7.31 (s, 3H), 7.07 (d, J=9 Hz, 2H), 5.10-4.90 (m, 2H),
4.80-4.58 (m, 2H), 3.22 (septet, J=7 Hz, 1H), 1.32 (d, J=7 Hz, 6H).
LRMS m/z 623 (M+H).sup.+.
23b)
6-[4-({[3-{[(2,6-Dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl)-4--
isoxazolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00190##
[0710] To a solution of ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate (6 mg, 0.01 mmol)
in 1:1 tetrahydrofuran:methanol (0.5 mL) was added 1 N sodium
hydroxide (0.020 mL, 0.020 mmol). The solution was heated in a
microwave reactor at 90.degree. C. for approximately 10 minutes and
then 1 N hydrochloric acid (0.020 mL, 0.020 mmol) was added. The
solution was concentrated and then dichloromethane and water were
added. The layers were separated and the organic layer was dried
over magnesium sulfate and concentrated to afford
6-[4-({[3-{[(2,6-dichlorophenyl)sulfinyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid (6 mg, 100%).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.38 (d, J=8 Hz, 1H),
8.23 (d, J=8 Hz, 2H), 8.04-8.01 (m, 2H), 7.68 (d, J=9 Hz, 2H), 7.31
(s, 3H), 7.07 (d, J=9 Hz, 2H), 5.14-4.99 (m, 2H), 4.84-4.59 (m,
2H), 3.24 (septet, J=7 Hz, 1H), 1.35 (d, J=7 Hz, 6H). LRMS (APCI)
m/z 595 (M+H).sup.+.
Example 24
6-[4-({[3-{[(2,6-Dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4-isoxa-
zolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00191##
[0711] 24a) Ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate
##STR00192##
[0713] To a solution of ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)thio]methyl}-5-(1-methylethyl)-4-isoxazol-
yl]methyl}oxy)phenyl]-2-quinolinecarboxylate (43 mg, 0.071 mmol) in
dichloromethane at 0.degree. C. was added meta-chloroperoxybenzoic
acid (39 mg, 0.16 mmol) and the mixture was allowed to stir at
0.degree. C. for 30 minutes. Then an additional 20 mg of
meta-chloroperoxybenzoic acid was added and the mixture was allowed
to stir overnight while warming slowly to ambient temperature. The
next day aqueous sodium bicarbonate was added and the solution was
extracted twice with dichloromethane. The combined organic layers
were dried over magnesium sulfate, adsorbed onto silica gel and
purified by chromatography (silica gel, 0-100% ethyl acetate in
hexanes gradient elution). The resulting solid was diluted with
dichloromethane and washed with aqueous sodium bicarbonate, dried
over magnesium sulfate and concentrated to afford ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate (4 mg, 0.006
mmol). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.39-8.33 (m, 2H),
8.21 (d, J=9 Hz, 1H), 8.05-8.01 (m, 2H), 7.71 (d, J=9 Hz, 2H),
7.45-7.37 (m, 3H), 7.10 (d, J=9 Hz, 2H), 5.15 (s, 2H), 4.86 (s,
2H), 4.57 (q, J=7 Hz, 2H), 3.25 (septet, J=7 Hz, 1H), 1.50 (t, J=7
Hz, 3H), 1.33 (d, J=7 Hz, 6H). LRMS (ESI) m/z 639 (M+H).sup.+.
24b)
6-[4-({[3-{[(2,6-Dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4--
isoxazolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00193##
[0715] To a solution of ethyl
6-[4-({[3-{[(2,6-dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylate (4 mg, 0.006 mmol)
in 1:1 tetrahydrofuran:methanol (0.5 mL) was added 1 N sodium
hydroxide (0.020 mL, 0.020 mmol). The solution was heated in a
microwave reactor at 75.degree. C. for approximately 10 minutes.
Then the mixture was diluted with dichloromethane and 1 N
hydrochloric (0.020 mL, 0.020 mmol) was added. The solution was
washed with water, dried over magnesium sulfate and concentrated to
afford
6-[4-({[3-{[(2,6-dichlorophenyl)sulfonyl]methyl}-5-(1-methylethyl)-4-isox-
azolyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid (1 mg, 25%).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 8.44 (d, J=8 Hz, 1H),
8.29 (d, J=8 Hz, 1H), 8.21 (d, J=8 Hz, 1H), 8.10-8.07 (m, 2H), 7.71
(d, J=8 Hz, 2H), 7.47-7.38 (m, 3H), 7.12 (d, J=8 Hz, 2H), 5.16 (s,
2H), 4.86 (s, 2H), 3.25 (septet, J=7 Hz, 1H), 1.34 (d, J=7 Hz, 6H).
LRMS m/z 609 [M-H].sup.-.
Example 25
6-[4-({[3-(3,5-Dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methyl-
}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00194##
[0716] 25a)
3-(3,5-Dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxylic
acid
##STR00195##
[0718] N-chlorosuccinimide (1.36 g, 10.2 mmol) was added to a
stirred solution of 3,5-dichloro-4-pyridinecarbaldehyde oxime (1.94
g, 10.2 mmol) in dimethylformamide (8 mL) and the solution was
heated in a 65.degree. C. oil bath for 1.5 hours. The solution was
poured into water and extracted with ether. The organic layer was
dried with MgSO.sub.4, filtered and concentrated to yield a crude
carboximidoyl chloride. A solution of methylisobutyrylacetate (1.7
mL, 12.3 mmol) in THF (2.5 mL) was stirred at 0.degree. C. as 0.5 N
solution of sodium methoxide in methanol (24.6 mL, 12.3 mmol) was
added. The solution was allowed to stir for ten minutes before the
addition of the crude
3,5-dichloro-N-hydroxy-4-pyridinecarboximidoyl chloride in THF (8.1
mL). The solution was allowed to stir at room temperature
overnight. The solution was then concentrated and the residue was
partitioned between water and ethyl acetate. The organic layer was
dried with MgSO.sub.4, filtered and concentrated. The residue was
purified by chromatography (silica gel, hexane to 1:9 ethyl
acetate: hexanes). Fractions containing the intermediate were
combined and concentrated. The residue was azetroped with methanol
then was diluted with THF (11 mL) and methanol (5.5 mL). A 1 N
solution of sodium hydroxide (3.3 mL) was added and the solution of
heated to 100.degree. C. for 500 seconds in a microwave reactor.
The solution was neutralized with 1 N HCl and concentrated to yield
a white solid. The residue was slurried in water and filtered to
yield
3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxyli-
c acid (0.57 g, 18%). .sup.1H NMR (400 MHz, DMSO-d6): .delta. 13.39
(s, 1H), 8.81 (s, 2H), 3.82 (septet, J=7 Hz, 1H), 1.34 (d, J=7 Hz,
6H).
25b)
[3-(3,5-Dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methanol
##STR00196##
[0720] A solution of
3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxylic
acid (0.54 g, 1.8 mmol) in THF (9 mL), was stirred as triethylamine
(0.25 mL, 1.8 mmol) was added. The solution was cooled in ice bath
before the addition of a 1 N solution of isopropylchloroformate in
toluene (1.8 mL, 1.8 mmol). The solution was allowed to stir for 30
minutes before being filtered into a solution of sodium borohydride
(91 mg, 2.4 mmol) in water (0.62 mL). The mixture was allowed to
warm to room temperature and stir for 3 days. The mixture was
filtered and the filtrate was partitioned between brine and ethyl
acetate. The organic layer was dried with MgSO.sub.4, filtered and
concentrated. The residue was purified by chromatography (silica
gel, hexane to 2:3 ethyl acetate: hexanes) to provide
[3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]meth-
anol (0.32 g, 57% as 0.3 ethyl acetate). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 8.79 (s, 2H), 4.96 (t, J=5 Hz, 1H), 4.20 (d,
J=5 Hz, 2H), 3.35 (septet, J=7 Hz, 1H), 1.29 (d, J=7 Hz, 6H).
25c) Methyl
6-[4-({[3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate
##STR00197##
[0722] Methyl 6-(4-hydroxyphenyl)-2-quinolinecarboxylate (100 mg,
0.36 mmol),
[3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]metha-
nol (103 mg, 0.36 mmol), triphenyl phosphine (94 mg, 0.36 mmol),
diisopropyl azodicarboxylate (0.07 mL, 0.36 mmol) and
dichloromethane (3.6 mL) were placed in a microwave reaction tube,
sealed and heated in a microwave reactor to 100.degree. C. for 10
minutes. The solution was concentrated then slurried in a 3:7
mixture of acetone: hexane. The resulting off-white solid was
purified by chromatography (silica gel, 3:7 acetone: hexanes) to
provide methyl
6-[4-({[3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate (0.10 g, 52%). .sup.1-HNMR
(400 MHz, DMSO-d.sub.6): .delta. 8.81 (s, 2H), 8.55 (d, J=9 Hz,
1H), 8.29 (d, J=2 Hz, 1H), 8.18-8.10 (m, 3H), 7.74 (d, J=9 Hz, 2H),
6.89 (d, J=9 Hz, 2H), 4.97 (s, 2H), 3.94 (s, 3H), 3.49 (septet, 7
Hz, 1H), 1.34 (d, J=7 Hz, 6H). ESI-LCMS m/z 548 (M+H).sup.+.
25d)
6-[4-({[3-(3,5-Dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00198##
[0724] A solution of methyl
6-[4-({[3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate (98 mg, 0.18 mmol) in THF (1.8
mL) was placed in a microwave reaction tube. Methanol (0.9 mL) was
added followed by 1.0 N sodium hydroxide (0.27 mL, 0.27 mmol)
before the tube was sealed and heated at 100.degree. C. for 600
seconds. The solution was neutralized with 1 N HCl. The resulting
mixture was partitioned between brine and ethyl acetate. The
organic layer was separated and concentrated to yield
6-[4-({[3-(3,5-dichloro-4-pyridinyl)-5-(1-methylethyl)-4-isoxazo-
lyl]methyl}oxy)phenyl]-2-quinolinecarboxylic acid as a yellow
powder (0.08 g, 80% as 0.25 ethyl acetate). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 13.37 (br s, approximately 1H), 8.81 (s,
approximately 2H), 8.53 (d, J=9 Hz, 1H), 8.27 (s, 1H), 8.17-8.08
(m, 3H), 7.73 (d, J=9 Hz, 2H), 6.89 (d, J=9 Hz, 2H), 4.97 (s, 2H),
3.49 (septet. J=7 Hz, 1H), 1.34 (d, J=7 Hz, 6H). HRMS
C.sub.28H.sub.21N.sub.3O.sub.4Cl.sub.2 m/z 534.0982
(M+H).sup.+.sub.Cal; 534.0981 (M+H).sup.+.sub.Obs.
Example 26
6-(4-{[(3,5-Dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinolinecarbo-
xylic acid
##STR00199##
[0725] 26a) Ethyl
3-cyclopentyl-2-(cyclopentylcarbonyl)-3-oxopropanoate
##STR00200##
[0727] A solution of ethyl 3-cyclopentyl-3-oxopropanoate (2.00 g,
10.9 mmol) in THF (20.4 mL) was stirred as sodium hydride 60% oil
dispersion (501 mg, 12.5 mmol) was added. The resulting solution
was allowed to stir for 30 minutes before the addition of
cyclopentanecarbonyl chloride (1.32 mL, 10.9 mmol). The mixture was
then allowed to stir at room temperature overnight. The solution
was then partitioned between ether and brine. The organic layer was
dried with MgSO.sub.4, filtered and concentrated. The residue was
purified by chromatography (silica gel, hexane to 1:19 ethyl
acetate: hexanes). Fractions containing the product were combined
and concentrated to yield
3-cyclopentyl-2-(cyclopentylcarbonyl)-3-oxopropanoate (2.21 g,
72%). ESI-LCMS m/z 279 (M-H).sup.-.
26b) Ethyl 3,5-dicyclopentyl-4-isoxazolecarboxylate
##STR00201##
[0729] A solution of ethyl
3-cyclopentyl-2-(cyclopentylcarbonyl)-3-oxopropanoate (2.20 g, 7.84
mmol) in EtOH (6.4 mL) was stirred as water (1.2 mL) followed by
hydroxylamine hydrochloride (1.02 g, 14.7 mmol) was added. The
solution was then heated to reflux for 3 hours. The solution was
concentrated and the residue was partitioned between ether and
saturated aqueous NaHCO.sub.3 solution. The organic layer was
washed with brine, dried with MgSO.sub.4, filtered and concentrated
to yield ethyl 3,5-dicyclopentyl-4-isoxazolecarboxylate as a yellow
oil (1.58 g, crude yield 73%). .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. 4.23 (q, J=7 Hz, 2H), 3.78-3.72 (m, 1H), 3.45-3.37 (m, 1H),
2.00-1.92 (m, 4H), 1.83-1.49 (m, 12H), 1.26 (t, J=7 Hz, 3H).
ESI-LCMS m/z 278 (M+H).sup.+.
26c) (3,5-Dicyclopentyl-4-isoxazolyl)methanol
##STR00202##
[0731] A solution of ethyl 3,5-dicyclopentyl-4-isoxazolecarboxylate
(1.57 g, 5.67 mmol) in THF (15.7 mL) was stirred in an ice bath as
a 1.5 M solution of diisobutylaluminum hydride in toluene (8.1 mL,
12.1 mmol) was added dropwise. The solution was allowed to stir at
0.degree. C. for 30 minutes then at room temperature for 5.5 hours.
An additional portion of diisobutylaluminum hydride (1.6 mL, 2.4
mmol) was added and the solution was allowed to stir for 45
minutes. The solution was cooled in ice bath before the addition of
a 10% aqueous solution of Rochelle's salt. The resulting solution
was allowed to stir overnight. The solution was extracted with
ethyl acetate and the organic layer was dried with MgSO.sub.4,
filtered and concentrated. The residue was purified by
chromatography (silica gel, 3:7 ethyl acetate: hexanes) to provide
(3,5-dicyclopentyl-4-isoxazolyl)methanol (0.80 g, 57% as 0.15 ethyl
acetate). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 4.84 (s,
1H), 4.26 (d, J=4 Hz, 2H), 3.30-3.22 (m, overlapping H.sub.2O),
3.13-3.06 (m, 1H), 1.98-1.87 (m, 4H), 1.74-1.56 (m, 12H).
26d) Methyl
6-(4-{[(3,5-dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinolinecarb-
oxylate
##STR00203##
[0733] Methyl 6-(4-hydroxyphenyl)-2-quinolinecarboxylate (109 mg,
0.39 mmol), (3,5-dicyclopentyl-4-isoxazolyl)methanol (92 mg, 0.39
mmol), triphenyl phosphine (102 mg, 0.39 mmol), diisopropyl
azodicarboxylate (0.075 mL, 0.39 mmol) and dichloromethane (3.9 mL)
were placed in a microwave reaction tube, sealed and heated in a
microwave reactor to 100.degree. C. for 10 minutes. The solution
was concentrated and the residue was purified by chromatography
(silica gel, hexane to 1:1 ethyl acetate: hexanes). Fractions
containing the product were combined and concentrated. The residue
was recrystallized from ethyl acetate and filtered to yield methyl
6-(4-{[(3,5-dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinolinecarb-
oxylate (67 mg, 35%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
8.57 (d, J=9 Hz, 1H), 8.33 (s, 1H), 8.19 (s, 2H), 8.12 (d, J=8 Hz,
1H), 7.84 (d, J=9 Hz, 2H), 7.17 (d, J=9 Hz, 2H), 4.98 (s, 2H), 3.94
(s, 3H), 3.41-3.37 (m, 1H), 3.16 (m, 1H), 1.98-1.94 (m, 4H),
1.76-1.56 (m, 12H).
26e)
6-(4-{[(3,5-Dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinoline-
carboxylic acid
##STR00204##
[0735] A solution of methyl
6-(4-{[(3,5-dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinolinecarb-
oxylate (65 mg, 0.13 mmol) in THF (1.3 mL) was placed in a
microwave reaction tube. Methanol (0.7 mL) was added followed by
1.0 N sodium hydroxide (0.2 mL, 0.2 mmol) before the tube was
sealed and stirred at room temperature for 30 minutes. The mixture
was heated at 100.degree. C. for 10 minutes. The solution was
concentrated and the residue was suspended in water before
neutralization with 1 N HCl. The resulting mixture was filtered and
the sample was dried in a drying oven at 45.degree. C. under
reduced pressure with P.sub.2O.sub.5 to yield
6-(4-{[(3,5-dicyclopentyl-4-isoxazolyl)methyl]oxy}phenyl)-2-quinolinecarb-
oxylic acid (0.53 g, 84%). .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. 8.47 (d, J=9 Hz, 1H), 8.27 (s, 1H), 8.18-8.11 (m, 2H), 8.07
(d, J=9 Hz, 1H), 7.82 (d, J=9 Hz, 2H), 7.16 (d, J=9 Hz, 2H), 4.98
(s, 2H), 3.40-3.20 (m, overlapping H.sub.20), 3.16-3.12 (m, 1H),
1.99 (m, 4H), 1.76-1.56 (m, 12H). HRMS
C.sub.301.sup.-1.sub.30N.sub.2O.sub.4 m/z 483.2278,
(M+H.sup.+.sub.Cal; 483.2277 (M+H).sup.+.sub.Obs.
Example 27
6-[4-({[3-[(Cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methyl}o-
xy)phenyl]-2-quinolinecarboxylic acid
##STR00205##
[0736] 27a) Methyl
3-(bromomethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate
##STR00206##
[0738] A solution of methyl
3-{[(1,1-dimethylethyl)oxy]methyl}-5-(1-methylethyl)-4-isoxazolecarboxyla-
te (2.00 g, 7.83 mmol) in dichloromethane (32.5 mL) was stirred at
0.degree. C. as trifluoroacetic acid (32.5 mL, 0.42 mol) was added
dropwise. The resulting solution was allowed to stir at 0.degree.
C. for 5 minutes then at room temperature for 30 minutes. The
solution was concentrated and the residue was partitioned between
ethyl acetate and saturated aqueous NaHCO.sub.3. The organic layer
was dried with MgSO.sub.4, filtered and concentrated. The residue
was purified by chromatography (silica gel, hexane to 2:3 ethyl
acetate: hexanes). Fractions containing methyl
3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate were
combined and concentrated. A solution of methyl
3-(hydroxymethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate (1.042
g, 5.23 mmol) in dichloromethane (24.3 mL) was stirred as carbon
tetrabromide (1.85 g, 5.5 mmol) was added. The solution was cooled
in ice bath before the addition of triphenyl phosphine (1.44 g, 5.5
mmol) in three portions. The solution was allowed to stir at
0.degree. C. for 30 minutes then at room temperature for 1.5 hours.
Hexane (67 mL) was added before the mixture was filtered through
Celite.RTM. and concentrated. The residue was purified by
chromatography (silica gel, hexane to 1:4 ethyl acetate: hexanes).
Fractions containing the product were combined and concentrated to
yield methyl
3-(bromomethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate (0.809 g,
57% as 0.10 ethyl acetate). .sup.1H NMR (400 MHz, DMSO-d.sub.6):
.delta. 4.70 (s, 2H), 3.82 (s, 3H), 3.69 (septet, J=7 Hz, 1H), 1.26
(d, J=7 Hz, 6H). ESI-LCMS m/z 262 (M+H).sup.+.
27b) Cyclopentyl
3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolecarboxylate
##STR00207##
[0740] A solution of cyclopentanol (0.44 mL, 4.8 mmol), in THF (10
mL), was stirred at 0.degree. C. as sodium hydride 60% oil
dispersion (192 mg, 4.8 mmol) was added. The solution was allowed
to stir for 30 minutes. A solution of methyl
3-(bromomethyl)-5-(1-methylethyl)-4-isoxazolecarboxylate (500 mg,
1.91 mmol), in THF (5 mL) was then added and the resulting solution
was allowed to warm to room temperature and stir for 3 hours. The
solution was then partitioned between brine and ethyl acetate. The
organic layer was dried with MgSO.sub.4, filtered and concentrated.
The residue purified by chromatography (silica gel, hexane to 1:4
ethyl acetate: hexanes). Fractions containing the product were
combined and concentrated to yield cyclopentyl
3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolecarboxylate
(94 mg, 15%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
5.29-5.26 (m, 1H), 4.54 (s, 2H), 3.98 (m, 1H), 3.64 (septet, 7 Hz,
1H), 1.85 (m, 2H), 1.72-1.56 (m, 12H), 1.47-1.40 (m, 2H), 1.25 (d,
J=7 Hz, 6H). ESI-LCMS m/z 322 (M+H).sup.+.
27c)
[3-[(Cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methanol
##STR00208##
[0742] A solution of cyclopentyl
3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolecarboxylate
(0.093 g, 0.29 mmol) in THF (2.8 mL) was stirred in an ice bath as
a 1.5M solution of diisobutylaluminum hydride in toluene (0.42 mL,
0.63 mmol) was added dropwise. The solution was allowed to stir at
room temperature for 2 hours. The solution was cooled to 0.degree.
C. before an additional portion of diisobutylaluminum hydride (0.42
mL, 0.63 mmol) was added and the solution was allowed to stir at
room temperature for one hour. A 10% aqueous solution of Rochelle's
salt (30mL) was added and the solution was allowed to stir for 20
hours. The solution was extracted with ethyl acetate and the
organic layer was dried with MgSO.sub.4, filtered and concentrated.
The residue was purified by chromatography (silica gel, hexane to
1:1 ethyl acetate: hexanes) to provide
[3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methanol
(0.051 g, 74%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 4.82
(t, J=5 Hz, 1H), 4.42 (s, 2H), 4.30 (d, J=5 Hz, 2H), 3.95(m, 1H)
3.24 (septet, J=7 Hz, overlapping H.sub.2O), 1.64-1.55 (m, 6H),
1.46 (m, 2H), 1.21 (d, J=7 Hz, 6H).
27d) Methyl
6-[4-({[3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylate
##STR00209##
[0744] A solution of
[3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methanol
(50 mg, 0.21 mmol) in dichloromethane (2.1 mL) was added to a
mixture of methyl 6-(4-hydroxyphenyl)-2-quinolinecarboxylate (57
mg, 0.21 mmol) and triphenyl phosphine (82 mg, 0.31 mmol) in a
microwave reaction tube. Diisopropyl azodicarboxylate (60 .mu.L,
0.31 mmol) was added before the tube was sealed and heated for 20
minutes at 100.degree. C. The mixture was concentrated and the
residue was purified by chromatography (silica gel, hexane to 2:3
ethyl acetate: hexanes). Fractions containing the product were
combined and concentrated. The residue was purified by
chromatography (silica gel, 10:9:1 hexanes: dichloromethane: ethyl
acetate) to provide methyl
6-[4-({[3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylate (11 mg, 11% as 0.10 ethyl
acetate). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.37-8.32 (m,
2H), 8.22 (d, J=9 Hz, 1H), 8.05-8.00 (m, 2H), 7.69 (d, J=9 Hz, 2H),
7.08 (d, J=9 Hz, 2H), 4.99 (s, 2H), 4.58 (s, 2H), 4.09 (s, 3H),
3.99-3.97 (m, 1H), 3.27 (septet, J=7 Hz, 1H), 1.70-1.59 (m, 6H),
1.49-1.46 (m, 2H), 1.33 (d, J=7 Hz, 6H).
27e)
6-[4-({[3-[(Cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]met-
hyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00210##
[0746] A solution of methyl
6-[4-({[3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylate (11 mg, 0.02 mmol) in THF (0.22
mL) was placed in a microwave reaction tube. Methanol (0.11 mL) was
added followed by 1 N sodium hydroxide (0.05 mL, 0.05 mmol). The
tube was sealed and heated to 100.degree. C. for 800 seconds. The
solution was concentrated then neutralized with 1 N HCl. Water was
added and the mixture was filtered to yield a bright yellow solid.
The sample was dried in a vacuum oven under reduced pressure at
45.degree. C. with P.sub.2O.sub.5. The sample was removed from the
oven to yield
6-[4-({[3-[(cyclopentyloxy)methyl]-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylic acid (10 mg, 95%). .sup.1H NMR
(400 MHz, DMSO-d.sub.6): .delta. 8.37 (d, J=9 Hz, 1H), 8.20-8.17
(m, 2H), 8.08-8.03 (m, 2H), 7.80 (d, J=9 Hz, 2H), 7.14 (d, J=9 Hz,
2H), 5.01 (s, 2H), 4.49 (s, 2H), 3.95-3.93 (m, 1H), 3.40-3.20 (br
s, 1H overlapped by water), 1.60-1.37 (m, 8H), 1.24 (d, J=7 Hz,
6H). HRMS C.sub.29H.sub.30N.sub.2O.sub.5 m/z 487.2228,
(M+H).sup.+.sub.Cal; 487.2227 (M+H).sup.+.sub.Obs.
Example 28
6-[4-({[3-(2,4-Dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methyl-
}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00211##
[0747] 28a) 2,4-Dichloro-3-pyridinecarbaldehyde
##STR00212##
[0749] A solution of diisopropylamine (2.6 mL, 18.5 mmol) in THF
(18.5 mL) was stirred at -78.degree. C. as a 2.5 N solution of
n-butyl lithium (7.4 mL, 18.5 mmol) was added. The solution was
stirred at 0.degree. C. for 30 minutes, then cooled to -78.degree.
C. for 30 minutes before the addition of 2,4-dichloropyridine (2
mL, 18.5 mmol). The solution was stirred at -78.degree. C. for
another 30 minutes before the addition of methyl formate (1.14 mL,
18.5 mmol). The solution was stirred for 30 minutes then was
partitioned between ethyl acetate and saturated aqueous
NaHCO.sub.3. The organic layer was dried with MgSO.sub.4, filtered
and concentrated. The residue was purified by chromatography
(silica gel, hexane to 3:7 ethyl acetate: hexanes) to provide
2,4-dichloro-3-pyridinecarbaldehyde (2.18 g, 67%). .sup.1H NMR (400
MHz, DMSO-d.sub.6): .delta. 10.29 (s, 1H), 8.54 (d, J=5 Hz, 1H),
7.75 (d, J=5 Hz, 1H).
28b) 2,4-Dichloro-3-pyridinecarbaldehyde oxime
##STR00213##
[0751] A solution of 2,4-dichloro-3-pyridinecarbaldehyde (2.15 g,
12.4 mmol) in ethanol (18 mL) was added to a solution of
hydroxylamine hydrochloride (1.01 g, 14.5 mmol) and sodium
hydroxide (0.58 g, 14.5 mL) in water (9 mL). The resulting solution
was heated in a 65.degree. C. oil bath for 3 hours. The mixture was
filtered to yield a white solid which was dried in a vacuum oven at
45.degree. C. with P.sub.2O.sub.5 to yield
2,4-dichloro-3-pyridinecarbaldehyde oxime (1.89 g, 80%). .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. 12.00 (s, 1H), 8.35 (d, J=5
Hz, 1H), 8.19 (s, 1H), 7.68 (d, J=5 Hz, 1H).
28c) 2,4-Dichloro-N-hydroxy-3-pyridinecarboximidoyl chloride
##STR00214##
[0753] A solution of 2,4-dichloro-3-pyridinecarbaldehyde oxime
(1.88 g, 9.9 mmol) in DMF (7.9 mL) was stirred as
N-chlorosuccinimide (1.32 g, 9.9 mmol) was added. The solution was
stirred in a 64.degree. C. oil bath for 1 hour then allowed to cool
to room temperature. The solution was poured onto ice. The solution
was extracted with ether and the organic layer was washed with
brine, dried with MgSO.sub.4, filtered and concentrated to yield
2,4-dichloro-N-hydroxy-3-pyridinecarboximidoyl chloride (1.94 g,
87%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 12.89 (s, 1H),
8.52 (d, J=5 Hz, 1H), 7.81 (d, J=5 Hz, 1H).
28d) Methyl
3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxylate
##STR00215##
[0755] A solution of methyl isobutyrylacetate (1.37 mL, 10.2 mmol)
in THF (2.1 mL) was stirred at 0.degree. C. as a 0.5 N solution of
sodium methoxide in methanol (20.3 mL, 10.2 mmol) was added. A
solution of 2,4-dichloro-N-hydroxy-3-pyridinecarboximidoyl chloride
(1.94 g, 8.64 mmol) in THF (19 mL) was added and the resulting
solution was allowed to warm to room temperature and stir
overnight. The mixture was concentrated and the residue partitioned
between ethyl acetate and brine. The organic layer was dried with
MgSO.sub.4, filtered and concentrated. The residue was purified by
chromatography (silica gel, hexanes to 3:7 ethyl acetate: hexanes
gradient elution) and fractions containing the product
concentrated. The residue was azetroped with methanol then
dichloromethane to provide methyl
3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxylate
(1.64 g, 66% as 0.15 dichloromethane). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 8.65 (d, J=5 Hz, 1H), 7.82 (d, J=5 Hz, 1H),
3.79 (septet, J=7 Hz, 1H), 3.63 (s, 3H), 1.34 (m, 6H).
28e)
[3-(2,4-Dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methanol
##STR00216##
[0757] A solution of methyl
3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxylate
(1.32 g, 4.2 mmol) in THF (11.7 mL) was stirred at a 0.degree. C.
as a 1.5 M solution of diisopropylaluminum hydride in toluene (9
mL, 13.4 mmol) was added; the solution was allowed to warm to room
temperature and stirred overnight. A 10% aqueous solution of
Rochelle's salt was added and the solution was then allowed to stir
for three hours. The layers were separated and the organic layer
was washed with brine. The organic layer was dried with MgSO.sub.4,
filtered and concentrated. The residue was purified by
chromatography (silica gel, hexane to ethyl acetate gradient
elution) to yield
[3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methanol
(1.20 g, 94% 0.2 ethyl acetate). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 8.52 (d, J=5 Hz, 1H), 7.78 (d, J=5 Hz, 1H),
4.94 (t, J=5 Hz, 1H), 4.20 (d, J=5 Hz, 2H), 3.34 (septet, J=7 Hz,
overlapping water), 1.29 (d, J=7 Hz, 6H).
28f) Methyl
6-[4-({[3-(2,4-dichloro-3-pyridinyl-5-(1-methylethyl-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinolinecarboxylate
##STR00217##
[0759] A solution of
[3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methanol
(55 mg, 0.19 mmol) in dichloromethane (1.9 mL) was added to a
mixture of methyl 6-(4-hydroxyphenyl)-2-quinolinecarboxylate (54
mg, 0.19 mmol) and triphenyl phosphine (50 mg, 0.19 mmol) in a
microwave reaction tube. Diisopropyl azodicarboxylate (37 .mu.L,
0.19 mmol) was added before the tube was sealed and heated for 10
minutes at 100.degree. C. The mixture was concentrated and the
residue was purified by chromatography (silica gel, hexane to 7:3
ethyl acetate: hexanes). Fractions containing the product were
combined and concentrated to yield the product (14 mg). Fractions
containing the product and other impurities were combined and the
residue was chromatographed (silica gel, dichloromethane to 1:19
methanol: dichloromethane) to provide the still impure product (20
mg). A second reaction was run according to the above procedure
with the following modifications: the scale of the reaction was
doubled for all reagents except 102 .mu.L of diisopropyl
azodicarboxylate was used; the reaction was then heated to
100.degree. C. for 20 minutes. The mixture was concentrated and
combined with the impure product from the first reaction (20 mg).
The sample was purified by chromatography (silica gel, hexane to
7:3 ethyl acetate: hexanes). Fractions containing the product were
combined and concentrated. The residue was slurried in methanol to
yield methyl
6-[4-({[3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate (87 mg, 32% combined yield)
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.56-8.51 (m, 2H),
8.29 (s, 1H), 8.18-8.10 (m, 3H), 7.80 (d, J=5 Hz, 1H), 7.74 (d, J=9
Hz, 2H), 6.92 (d, J=9 Hz, 2H), 4.96 (s, 2H), 3.94 (s, 3H), 3.49
(septet, J=7 Hz, 1H), 1.34 (d, J=7 Hz, 6H).
28 g)
6-[4-({[3-(2,4-Dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]-
methyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00218##
[0761] A solution of methyl
6-[4-({[3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate (85 mg, 0.16 mmol) (from
multiple batches) in THF (1.6 mL) was stirred as a 1 N aqueous
sodium hydroxide solution (0.24 mL, 0.24 mmol) was added. The
solution was allowed to stir at room temperature overnight. The
solution was neutralized with 1 N HCl and concentrated. The residue
was partitioned between ethyl acetate and brine. The organic layer
was dried with MgSO.sub.4. The MgSO.sub.4 was slurried in
dichloromethane, filtered and the combined filtrates were
concentrated to yield
6-[4-({[3-(2,4-dichloro-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylic acid as a bright yellow foam
(66 mg, 76% 0.12 ethyl acetate). .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 13.37 (s, 1H), 8.54-8.51 (m, 2H), 8.28 (d,
J=1 Hz, 1H), 8.17-8.08 (m, 3H), 7.80 (d, J=5 Hz, 1H), 7.74 (d, J=9
Hz, 2H), 6.92 (d, J=9 Hz, 2H), 4.96 (s, 2H), 3.49 (sept, J=7 Hz,
1H), 1.34 (d, J=7 Hz, 6H). HRMS
C.sub.28H.sub.21N.sub.3O.sub.4Cl.sub.2 m/z 534.0982,
(M+H).sup.+.sub.Cal; 534.0980 (M+H).sup.+.sub.Obs.
Example 29
6-[4-({[3-(2,4-Dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methyl-
}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00219##
[0762] 29a) 2,4-Dimethyl-3-pyridinecarbaldehyde oxime
##STR00220##
[0764] A solution of hydroxylamine hydrochloride (0.21 g, 3.0 mmol)
and sodium hydroxide (0.12 g, 3.0 mmol) in water (1.9 mL) was added
to a solution of 2,4-dimethyl-3-pyridinecarbaldehyde (0.35 g, 2.6
mmol) in ethanol (3.8 mL). The solution was heated in a 65.degree.
C. oil bath for 2 hours. The mixture was concentrated and the
residue diluted with water and filtered. The resulting white solid
was dried in a 45.degree. C. vacuum oven with P.sub.2O.sub.5 to
yield 2,4-dimethyl-3-pyridinecarbaldehyde oxime (345 mg, 88%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 11.44 (s, 1H), 8.33
(s, 1H), 8.23 (d, J=5 Hz, 1H), 7.08 (d, J=5 Hz, 1H), 2.48 (s,
overlapping DMSO), 2.33 (s, 3H).
29b) Methyl
3-(2,4-dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxylate
##STR00221##
[0766] A solution of 2,4-dimethyl-3-pyridinecarbaldehyde oxime (341
mg, 2.27 mmol) in DMF (1.8 mL) was stirred as N-chlorosuccinimide
(303 mg, 2.27 mmol) was added. The solution was stirred in a
65.degree. C. oil bath for 3 hours. The solution was allowed to
cool before being poured on ice. The solution was extracted with
ether. The organic layer containing the crude
N-hydroxy-2,4-dimethyl-3-pyridinecarboximidoyl chloride was dried
with MgSO.sub.4, filtered and concentrated.
[0767] A 0.5 N solution of sodium methoxide in methanol (2.8 mL,
1.4 mmol) was added to a solution of methyl isobutyrylacetate (0.19
mL, 1.34 mmol) in THF (0.24 mL) at 0.degree. C. A solution of the
crude N-hydroxy-2,4-dimethyl-3-pyridinecarboximidoyl chloride (0.22
g, 1.2 mmol) in THF (2.2 mL) was added and the resulting mixture
was allowed to warm to room temperature and stir overnight. The
solution was concentrated and the residue was partitioned between
ethyl acetate and brine. The organic layer was dried with
MgSO.sub.4, filtered and concentrated. The residue was
chromatographed (silica gel, hexane to 3:2 ethyl acetate: hexane)
to provide methyl
3-(2,4-dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolecarboxylate
(0.17 g, 52%). ESI-LCMS m/z 275 (M+H).sup.-.
29c)
[5-(2,4-Dimethyl-3-pyridinyl)-2-(1-methylethyl)-1,4-cyclopentadien-1--
yl]methanol
##STR00222##
[0769] A solution of methyl
5-(2,4-dimethyl-3-pyridinyl)-2-(1-methylethyl)-1,4-cyclopentadiene-1-carb-
oxylate (0.17 g, 0.62 mmol) in THF (1.7 mL) was stirred at
0.degree. C. as a 1.5 M solution of diisopropylaluminum hydride in
toluene (0.88 mL, 1.32 mmol) was added. The solution was allowed to
warm to room temperature and stir overnight. The solution was then
cooled to 0.degree. C. before adding additional portion of
diisopropylaluminum hydride in toluene (0.88 mL, 1.32 mmol). The
solution was allowed to stir overnight before the addition a 10%
aqueous solution of Rochelle's salt. The solution was then allowed
to stir overnight. The solution was extracted with ethyl acetate
and the organic layer was dried with MgSO.sub.4, filtered and
concentrated. The residue was purified by chromatography (silica
gel, dichloromethane to 19:1 dichloromethane: methanol) to yield
[5-(2,4-dimethyl-3-pyridinyl)-2-(1-methylethyl)-1,4-cyclopentadien-1-yl]m-
ethanol (98 mg, 64%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
8.37 (d, J=5 Hz, 1H), 7.18 (d, J=5 Hz, 1H), 4.87 (t, J=5 Hz, 1H),
4.04 (d, J=5 Hz, 2H), 3.34 (septet, J=7 Hz, overlapping water),
2.18 (s, 3H), 2.02 (s, 3H), 1.30 (d, J=7 Hz, 6H).
29d) Methyl
6-[4-({[3-(2,4-dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate
##STR00223##
[0771] A mixture of
[5-(2,4-dimethyl-3-pyridinyl)-2-(1-methylethyl)-1,4-cyclopentadien-1-yl]m-
ethanol (96 mg, 0.39 mmol), methyl
6-(4-hydroxyphenyl)-2-quinolinecarboxylate (0.14 g, 0.39 mmol),
triphenyl phosphine (0.10 g, 0.39 mmol), and diisopropyl
azodicarboxylate (0.10 mL, 0.39 mmol) in dichloromethane (3.9 mL)
was heated in a microwave reactor at 100.degree. C. for 20 minutes.
The solution was concentrated and the residue was purified by
chromatography (silica gel, hexane to ethyl acetate). Fractions
containing the product were combined and concentrated. The residue
was recrystalized from ethyl acetate and hexane to yield methyl
6-[4-({[3-(2,4-dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate (22 mg, 11%). .sup.1H NMR (400
MHz, DMSO-d.sub.6): .delta. 8.55 (d, J=9 Hz, 1H), 8.36 (d, J=5 Hz,
1H), 8.28 (s, 1H), 8.18-8.10 (m, 3H), 7.74 (d, J=9 Hz, 2H), 7.19
(d, J=5 Hz, 1H), 6.96 (d, J=9 Hz, 2H), 4.79 (s, 2H), 3.94 (s, 3H),
3.47 (septet, J=7 Hz, 1H), 2.23 (s, 3H), 2.07 (s, 3H), 1.33 (d, J=7
Hz, 6H). ESI-LCMS m/z 508 (M+H).sup.+.
29e)
6-[4-({[3-(2,4-Dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-2-quinolinecarboxylic acid
##STR00224##
[0773] A solution of methyl
6-[4-({[3-(2,4-dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl]methy-
l}oxy)phenyl]-2-quinolinecarboxylate (20 mg, 0.04 mmol) in THF (0.4
mL) was placed in a microwave reaction tube followed by methanol
(0.20 mL) followed by a 1 N solution of sodium hydroxide (0.08 mL,
0.08 mmol). The tube was sealed and heated to 100.degree. C. for 10
minutes. The solution was neutralized with 1 N HCl and
concentrated. The residue was diluted with water and filtered. The
resulting solid was dried with P.sub.2O.sub.5 in a 45.degree. C.
vacuum oven under reduced pressure to yield
6-[4-({[3-(2,4-dimethyl-3-pyridinyl)-5-(1-methylethyl)-4-isoxazolyl-
]methyl}oxy)phenyl]-2-quinolinecarboxylic acid (17 mg, 86%).
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 13.5-13.2 (br s, 1H),
8.53 (d, J=9 Hz, 1H), 8.38 (d, J=5 Hz, 1H), 8.27 (s, 1H), 8.17-8.08
(m, 3H), 7.74 (d, J=9 Hz, 2H), 7.22 (d, J=5 Hz, 1H), 7.22 (d, J=9
Hz, 2H), 4.79 (s, 2H), 3.47 (septet, J=7 Hz, 1H), 2.24 (s, 3H),
2.08 (s, 3H), 1.33 (d, J=7 Hz, 6H). ESI-LCMS m/z 494
(M+H).sup.+.
Example 30
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-7-fluoro-2-quinolinecarboxylic acid
##STR00225##
[0774] 30a) 5-Bromo-4-fluoro-2-nitro-benzaldehyde
##STR00226##
[0776] 4-Fluoro-3-bromo-benzaldehyde (3.00 g, 14.78 mmol) was added
to a stirred solution of concentrated nitric acid (67%, 2.02 mL,
29.55 mmol) in concentrated sulfuric acid (18 mL) at 0.degree. C.
After the addition was complete, the ice bath was removed and the
reaction was allowed to stir for 3 hours at room temperature. Then,
the mixture was poured into ice and extracted with diethyl ether.
The organic layer was dried over anhydrous magnesium sulfate, then
filtered and concentrated. The residue was purified by silica gel
chromatography eluting with 1:9 ethyl acetate:hexanes to give 2.66
g (73%) of 5-bromo-4-fluoro-2-nitro-benzaldehyde as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 10.14 (s, 1H), 8.31
(d, J=8 Hz, 1H), 8.25 (d, J=7 Hz, 1H).
30b) Ethyl 6-bromo-7-fluoro-2-quinolinecarboxylate
##STR00227##
[0778] 5-Bromo-4-fluoro-2-nitro-benzaldehyde (1.50 g, 6.05 mmol)
and ethyl pyruvate (737.4 mg, 6.35 mmol) in ethanol (15 mL) was
added via canula to zinc(II) chloride (4.12 g, 30.24 mmol), tin(II)
chloride (5.73 g, 30.24 mmol), and activated 4 .ANG. molecular
sieve pellets (1.50 g) in ethanol (15 mL) under argon. The mixture
was heated at 70.degree. C. in an oil bath for four hours, then
allowed to cool to room temperature, and carefully quenched with
saturated sodium bicarbonate. Ethyl acetate was added and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:6
ethyl acetate:hexanes to give 368.5 mg (20%) of ethyl
6-bromo-7-fluoro-2-quinolinecarboxylate as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 8.63 (d, J=8 Hz, 1H), 8.58 (d, J=9
Hz, 1H), 8.15 (s, 1H), 8.13 (s, 1H), 4.42 (q, J=7 Hz, 2H), 1.37 (t,
J=7 Hz, 3H). ESI-LCMS m/z 297 (M+H).sup.+.
30c) Ethyl 7-fluoro-6-(4-hydroxyphenyl)-2-quinolinecarboxylate
##STR00228##
[0780] Palladium(II) acetate (8.2 mg, 36.5 .mu.mol) was added to
ethyl 6-bromo-7-fluoro-2-quinolinecarboxylate (217.8 mg, 730.6
.mu.mol), 4-hydroxy-phenyl-boronic acid (246.0 mg, 1.10 mmol),
triphenylphosphine (19.2 mg, 73.1 .mu.mol), and potassium phosphate
(542.8 mg, 2.56 mmol). Then, dioxane (3.6 mL) was added to the
mixture, followed by water (73 .mu.L), and the reaction mixture was
heated open to the atmosphere at 60.degree. C. in an oil bath for
14 hours, then allowed to cool to room temperature. Water was added
followed by ethyl acetate and the mixture was filtered through
Celite.RTM.. The filtrate was extracted with ethyl acetate and the
organic layer was dried over anhydrous magnesium sulfate, then
filtered and concentrated. The residue was purified by silica gel
chromatography eluting with 2:3 ethyl acetate:hexanes to give 138.9
mg (61%) of ethyl
7-fluoro-6-(4-hydroxyphenyl)-2-quinolinecarboxylate as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 9.82 (s, 1H), 8.61 (d,
J=9 Hz, 1H), 8.23 (d, J=9 Hz, 1H), 8.11 (d, J=8 Hz, 1H), 8.01 (d,
J=12 Hz, 1H), 7.53 (d, J=8 Hz, 2H), 6.92 (d, J=8 Hz, 2H), 4.42 (q,
J=7 Hz, 2H), 1.38 (t, J=7 Hz, 3H). ESI-LCMS m/z 312
(M+H).sup.+.
30d) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-7-fluoro-2-quinolinecarboxylate
##STR00229##
[0782] Cesium carbonate (62.6 mg, 192.2 .mu.mol) was added to ethyl
7-fluoro-6-(4-hydroxyphenyl)-2-quinolinecarboxylate (54.4 mg, 174.7
.mu.mol) and
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(58.6 mg, 192.2 .mu.mol) in N,N-dimethylformamide (1.7 mL) at room
temperature. The resulting suspension was stirred for 68 hours,
then water was added and the reaction mixture was extracted with
ethyl acetate. The organic layer was dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 3:7 ethyl acetate:hexanes
to give 49.4 mg (49%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-7-fluoro-2-quinolinecarboxylate as an oil. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 8.61 (d, J=9 Hz, 1H), 8.23 (d, J=9 Hz,
1H), 8.11 (d, J=9 Hz, 1H), 8.02 (d, J=12 Hz, 1H), 7.65-7.62 (m,
2H), 7.56 (d, J=7 Hz, 2H), 7.56-7.52 (m, 1H), 6.96 (d, J=9 Hz, 2H),
4.90 (s, 2H), 4.42 (q, J=7 Hz, 2H), 2H), 3.48 (septet, J=7 Hz, 1H),
1.38 (t, J=7 Hz, 3H), 1.34 (d, J=7 Hz, 6H). ESI-LCMS m/z 579
(M+H).sup.+.
30e)
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-7-fluoro-2-quinolinecarboxylic acid
##STR00230##
[0784] A 1 N solution of sodium hydroxide (154.6 .mu.L, 154.6
.mu.mol) was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-7-fluoro-2-quinolinecarboxylate (44.8 mg, 77.3 .mu.mol) in
tetrahydrofuran and methanol (1:1, 1.6 mL) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(154.6 .mu.L), water added, and the reaction mixture was extracted
with ethyl acetate. The organic layer was washed with brine, then
dried over anhydrous magnesium sulfate, filtered and concentrated
to give 40.6 mg (95%) of
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-7-fluoro-2-quinolinecarboxylic acid as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 13.52 (br s, 1H), 8.56 (d, J=9 Hz,
1H), 8.21 (d, J=9 Hz, 1H), 8.09 (d, J=9 Hz, 1H), 7.98 (d, J=12 Hz,
1H), 7.65-7.61 (m, 2H), 7.56 (d, J=9 Hz, 2H), 7.56-7.52 (m, 1H),
6.96 (d, J=9 Hz, 2H), 4.90 (s, 2H), 3.48 (septet, J=7 Hz, 1H), 1.34
(d, J=7 Hz, 6H). HRMS C.sub.29H.sub.21Cl.sub.2FN.sub.2O.sub.4 m/z
551.0941 (M+H).sup.+.sub.Cal; 551.095 (M+H).sup.+.sub.Obs.
Example 31
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-3-methyl-2-quinolinecarboxylic acid
##STR00231##
[0785] 31a) 5-Bromo-2-nitro-benzaldehyde
##STR00232##
[0787] 3-Bromo-benzaldehyde (18.50 g, 99.99 mmol) was added to a
stirred solution of concentrated nitric acid (70%, 13.09 mL, 199.98
mmol) in concentrated sulfuric acid (125 mL) at 0.degree. C. After
the addition was complete, the ice bath was removed and the
reaction was allowed to stir for 5 hours at room temperature. Then,
the mixture was poured into ice and the solid was collected by
filtration. The filtrate was extracted with diethyl ether. The
organic layer was dried over anhydrous magnesium sulfate, then
filtered and concentrated. The residue was combined with the solid
previously obtained via the first filtration and dissolved in
diethyl ether. Hexanes were added and after crystallization 14.66 g
(64%) of 5-bromo-2-nitro-benzaldehyde was collected by filtration
and drying. .sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 10.19 (s,
1H), 8.10 (s, 2H), 8.02 (s, 1H).
31b) Ethyl 2-oxobutanoate
##STR00233##
[0789] Jones Reagent (2.67 M, 7.51 mL, 20.05 mmol) was added
dropwise to ethyl 2-hydroxybutanoate (2.65 g, 20.05 mmol) in
acetone (67 mL) at 0.degree. C., then the reaction mixture was
stirred for 10 minutes. Any excess Jones reagent was quenched by
the addition of iso-propanol, then the reaction mixture was
neutralized with saturated sodium bicarbonate, and the acetone was
removed by evaporation. Water was added and the solution was
extracted with diethyl ether. The organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:4
diethyl ether:hexanes to give 346.7 mg (13%) of ethyl
2-oxobutanoate as an oil. .sup.1H NMR (400 MHz, d.sub.6-DMSO):
.delta. 4.19 (q, J=7 Hz, 2H), 2.80 (q, J=7 Hz, 2H), 1.24 (t, J=7
Hz, 3H), 0.94 (t, J=7 Hz, 3H).
31c) Ethyl 6-bromo-3-methyl-2-quinolinecarboxylate
##STR00234##
[0791] A 2.2 M solution of zinc(II) chloride (6.05 mL, 13.32 mmol)
in diethyl ether was added to tin(II) chloride (2.53 g, 13.32 mmol)
and activated 4 .ANG. molecular sieve pellets (612.8 mg) under
argon. Then, 5-bromo-2-nitro-benzaldehyde (612.8 mg, 2.66 mmol) and
ethyl 2-oxobutanoate (346.7 mg, 2.66 mmol) in ethanol (13 mL) were
added to the reaction mixture via canula. The mixture was heated at
70.degree. C. in an oil bath for three hours, then allowed to cool
to room temperature, and carefully quenched with saturated sodium
bicarbonate. Ethyl acetate was added and the mixture was filtered
through Celite.RTM.. The filtrate was extracted with ethyl acetate
and the organic layer was dried over anhydrous magnesium sulfate,
then filtered and concentrated. The residue was purified by silica
gel chromatography eluting with 1:6 ethyl acetate:hexanes to give
137.9 mg (18%) of ethyl 6-bromo-3-methyl-2-quinolinecarboxylate as
a solid. .sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 8.30 (s, 1H),
8.27 (d, J=2 Hz, 1H), 7.99 (d, J=9 Hz, 1H), 7.88 (dd, J=9, 2 Hz,
1H), 4.41 (q, J=7 Hz, 2H), 2.54 (s, 3H), 1.35 (t, J=7 Hz, 3H).
ESI-LCMS m/z 294 (M+H).sup.+.
31d) Ethyl 6-(4-hydroxyphenyl)-3-methyl-2-quinolinecarboxylate
##STR00235##
[0793] Palladium(II) acetate (4.9 mg, 22.0 .mu.mol) was added to
ethyl 6-bromo-3-methyl-2-quinolinecarboxylate (129.6 mg, 440.6
.mu.mol), 4-hydroxy-phenyl-boronic acid (91.2 mg, 660.9 .mu.mol),
triphenylphosphine (11.6 mg, 44.0 .mu.mol), and potassium phosphate
(327.3 mg, 1.54 mmol). Then, dioxane (4.4 mL) was added to the
mixture, followed by water (44.1 .mu.L), and the reaction mixture
was heated open to the atmosphere at 60.degree. C. in an oil bath
for 17 hours, then allowed to cool to room temperature. Water was
added followed by ethyl acetate and the mixture was filtered
through Celite.RTM.. The filtrate was extracted with ethyl acetate
and the organic layer was dried over anhydrous magnesium sulfate,
then filtered and concentrated. The residue was purified by silica
gel chromatography eluting with 2:3 ethyl acetate:hexanes to give
66.4 mg (49%) of ethyl
6-(4-hydroxyphenyl)-3-methyl-2-quinolinecarboxylate as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO); .delta. 9.71 (s, 1H), 8.31 (s,
1H), 8.13 (s, 1H), 8.07-8.02 (m, 2H), 7.68 (d, J=9 Hz, ESI-LCMS m/z
308 (M+H).sup.+.
31e) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-3-methyl-2-quinolinecarboxylate
##STR00236##
[0795] Cesium carbonate (71.7 mg, 220.1 .mu.mol) was added to ethyl
6-(4-hydroxyphenyl)-3-methyl-2-quinolinecarboxylate (61.5 mg, 200.1
.mu.mol) and
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(67.0 mg, 220.1 .mu.mol) in N,N-dimethylformamide (4 mL) at room
temperature. The resulting suspension was stirred for 18 hours,
then water was added and the reaction mixture was extracted with
ethyl acetate. The organic layer was dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 3:7 ethyl acetate:hexanes
to give 50.7 mg (44%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-methyl-2-quinolinecarboxylate as an oil. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 8.32 (s, 1H), 8.14 (s, 1H), 8.08-8.01
(m, 2H), 7.72 (s, 1H), 7.70 (s, 1H), 7.63 (d, J=8 Hz, 2H), 7.55
(dd, J=9, 7 Hz, 1H), 6.93 (d, J=9 Hz, 2H), 4.89 (s, 2H), 4.41 (q,
J=7 Hz, 2H), 3.48 (septet, J=7 Hz, 1H), 2.55 (s, 3H), 1.36 (t, J=7
Hz, 3H), 1.34 (d, J=7 Hz, 6H). ESI-LCMS m/z 575 (M+H).sup.+.
31f)
6-[4-({[3-(2,6-Dichlorophenyl-5-(1-methylethyl-4-isoxazolyl]methyl}ox-
y)phenyl]-3-methyl-2-quinolinecarboxylic acid
##STR00237##
[0797] A 1 N solution of sodium hydroxide (150.5 .mu.L, 150.5
.mu.mol) was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-methyl-2-quinolinecarboxylate (43.3 mg, 75.2 .mu.mol) in
tetrahydrofuran and methanol (1:1, 1.5 mL) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(150.5 .mu.L), water was added, and the reaction mixture was
extracted with ethyl acetate. The organic layer was washed with
brine, then dried over anhydrous magnesium sulfate, filtered and
concentrated to give 40.0 mg (97%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-3-methyl-2-quinolinecarboxylic acid as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 13.44 (br s, 1H), 8.29 (s, 1H),
8.14 (s, 1H), 8.08-8.00 (m, 2H), 7.72 (s, 1H), 7.70 (s, 1H), 7.64
(d, J=8 Hz, 2H), 7.55 (dd, J=9, 7 Hz, 1H), 6.93 (d, J=9 Hz, 2H),
4.89 (s, 2H), 3.48 (septet, J=7 Hz, 1H), 2.57 (s, 3H), 1.34 (d, J=7
Hz, 6H). HRMS C.sub.30H.sub.24Cl.sub.2N.sub.2O.sub.4 m/z 547.1191
(M+H).sup.+.sub.Cal; 547.1196 (M+H).sup.+.sub.Obs.
Example 32
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)--
3-methylphenyl]-2-quinolinecarboxylic acid
##STR00238##
[0798] 32a)
2-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol
##STR00239##
[0800] Potassium acetate (7.08 g, 72.18 mmol) was added to
4-bromo-2-methyl-phenol (3.00 g, 16.04 mmol) in
N,N-dimethylformamide (40 mL) at room temperature under argon.
Then, 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane
(6.11 g, 24.06 mmol) was added to the reaction mixture, followed by
the addition of
dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(II)
dichloromethane adduct (654.9 mg, 802.0 .mu.mol), and the reaction
vessel was evacuated and purged with argon. The mixture was heated
at 80.degree. C. in an oil bath for sixty-eight hours, then allowed
to cool to room temperature, and concentrated. Water was added,
followed by diethyl ether, and the mixture was filtered through
Celite.RTM.. The filtrate was extracted with diethyl ether and the
organic layer was dried over anhydrous magnesium sulfate, then
filtered and concentrated. The residue was purified by silica gel
chromatography eluting with 1:6 ethyl acetate:hexanes to give 1.63
g (43%) of
2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol as a
oil. .sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 9.66 (s, 1H),
7.36 (s, 1H), 7.31 (d, J=8 Hz, 1H), 6.74 (d, J=8 Hz, 1H), 2.09 (s,
3H), 1.24 (s, 12H). ESI-LCMS m/z 233 (M-H).sup.-.
32b) (4-hydroxy-3-methylphenyl)boronic acid
##STR00240##
[0802] Sodium periodate (4.22 g, 19.74 mmol) was added to
2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol
(1.54 g, 6.58 mmol) and ammonium acetate (1.52 g, 19.74 mmol) in
acetone and water (2:1, 66 mL) at room temperature. The mixture was
stirred for nineteen hours, then filtered, and concentrated. Sodium
chloride was added to the filtrate, and it was extracted with ethyl
acetate. The organic layer was dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 1:1 ethyl acetate:hexanes
to ethyl acetate to give 364.4 mg (36%) of
(4-hydroxy-3-methylphenyl)boronic acid as a solid. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 9.31 (s, 1H), 7.55 (s, 1H), 7.52 (d,
J=8 Hz, 1H), 6.76 (d, J=8 Hz, 1H), 2.14 (s, 3H). ESI-LCMS m/z 151
(M-H).sup.-.
32c) Ethyl 6-bromo-2-quinolinecarboxylate
##STR00241##
[0804] Ethyl pyruvate (535.4 mg, 4.61 mmol) was added to
5-Bromo-2-nitro-benzaldehyde (1.01 g, 4.39 mmol) in ethanol (22 mL)
at room temperature under argon. Then, tin(II) chloride (4.16 g,
21.95 mmol) was added to the reaction mixture, followed by the
addition of zinc(II) chloride (2.99 g, 21.95 mmol), and finally
activated 4 .ANG. molecular sieve pellets (1.01 g). The mixture was
heated at 70.degree. C. in an oil bath for twenty hours, then
allowed to cool to room temperature, and carefully quenched with
saturated sodium bicarbonate. Ethyl acetate was added, and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate, and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with
dichloromethane and subsequently a second silica gel chromatography
purification eluting with 1:4 ethyl acetate:hexanes to give 173.6
mg (14%) of ethyl 6-bromo-2-quinolinecarboxylate as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 8.55 (d, J=9 Hz, 1H),
8.42 (d, J=2 Hz, 1H), 8.16 (d, J=9 Hz, 1H), 8.11 (d, J=9 Hz, 1H),
7.98 (dd, J=9, 2 Hz, 1H), 4.42 (q, J=7 Hz, 2H), 1.37 (t, J=7 Hz,
3H). ESI-LCMS m/z 280 (M+H).sup.+.
32d) Ethyl 6-(4-hydroxy-3-methylphenyl)-2-quinolinecarboxylate
##STR00242##
[0806] Palladium(II) acetate (4.7 mg, 21.0 .mu.mol) was added to
ethyl 6-bromo-2-quinolinecarboxylate (117.8 mg, 420.5 .mu.mol),
(4-hydroxy-3-methylphenyl)boronic acid (95.9 mg, 630.8 .mu.mol),
triphenylphosphine (11.0 mg, 42.1 .mu.mol), and potassium phosphate
(312.4 mg, 1.47 mmol). Then, dioxane (4.2 mL) was added to the
mixture, followed by water (42 .mu.L), and the reaction mixture was
heated open to the atmosphere at 60.degree. C. in an oil bath for
16 hours, then allowed to cool to room temperature. Water was
added, followed by ethyl acetate, and the mixture was filtered
through Celite.RTM.. The filtrate was extracted with ethyl acetate,
and the organic layer was dried over anhydrous magnesium sulfate,
then filtered and concentrated. The residue was purified by silica
gel chromatography eluting with 2:3 ethyl acetate:hexanes to give
51.4 mg (40%) of ethyl
6-(4-hydroxy-3-methylphenyl)-2-quinolinecarboxylate as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 9.64 (s, 1H), 8.55 (d,
J=9 Hz, 1H), 8.27 (s, 1H), 8.18-8.13 (m, 2H), 8.11 (d, J=8 Hz, 1H),
7.62 (s, 1H), 7.53 (dd, J=8, 2 Hz, 1H), 6.91 (d, J=8 Hz, 1H), 4.42
(q, J=7 Hz, 2H), 2.22 (s, 3H), 1.38 (t, J=7 Hz, 3H). ESI-LCMS m/z
308 (M+H).sup.+.
32e) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)-3-methylphenyl]-2-quinolinecarboxylate
##STR00243##
[0808] Cesium carbonate (54.9 mg, 168.6 .mu.mol) was added to ethyl
6-(4-hydroxy-3-methylphenyl)-2-quinolinecarboxylate (47.1 mg, 153.3
.mu.mol) and
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(51.3 mg, 168.6 .mu.mol) in N,N-dimethylformamide (3.1 mL) at room
temperature. The resulting suspension was stirred for twenty hours,
then water was added and the reaction mixture was extracted with
ethyl acetate. The organic layer was dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 3:7 ethyl acetate:hexanes
to give 19.1 mg (22%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-3-methylphenyl]-2-quinolinecarboxylate as an oil. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 8.56 (d, J=9 Hz, 1H), 8.28 (s, 1H),
8.19-8.09 (m, 3H), 7.66-7.59 (m, 4H), 7.55 (dd, J=9, 7 Hz, 1H),
7.09 (d, J=9 Hz, 1H), 4.91 (s, 2H), 4.42 (q, J=7 Hz, 2H), 3.52
(septet, J=7 Hz, 1H), 1.97 (s, 3H), 1.38 (t, J=7 Hz, 3H), 1.36 (d,
J=7 Hz, 6H). ESI-LCMS m/z 575 (M+H).sup.+.
32f)
6-[4-({[3-(2,6-Dichlorophenyl-5-(1-methylethyl-4-isoxazolyl]methyl}ox-
y)-3-methylphenyl]-2-quinolinecarboxylic acid
##STR00244##
[0810] A 1 N solution of sodium hydroxide (59.4 .mu.L, 59.4
.mu.mol) was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-3-methylphenyl]-2-quinolinecarboxylate (17.1 mg, 29.7 .mu.mol) in
tetrahydrofuran and methanol (1:1, 594.3 .mu.L) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(59.4 .mu.L), water was added, and the reaction mixture was
extracted with ethyl acetate. The organic layer was washed with
brine, then dried over anhydrous magnesium sulfate, filtered and
concentrated to give 16.1 mg (99%) of
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]me-
thyl}oxy)-3-methylphenyl]-2-quinolinecarboxylic acid as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 13.37 (br s, 1H), 8.53
(d, J=8 Hz, 1H), 8.28 (s, 1H), 8.18-8.12 (m, 2H), 8.10 (d, J=8 Hz,
1H), 7.66-7.58 (m, 4H), 7.55 (dd, J=9, 7 Hz, 1H), 7.09 (d, J=9 Hz,
1H), 4.91 (s, 2H), 3.52 (septet, J=7 Hz, 1H), 1.98 (s, 3H), 1.36
(d, J=7 Hz, 6H). HRMS C.sub.30H.sub.24Cl.sub.2N.sub.2O.sub.4 m/z
547.1191 (M+H).sup.+.sub.Cal; 547.1198 (M+H).sup.+.sub.Obs.
Example 33
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)--
2-methylphenyl]-2-quinolinecarboxylic acid
##STR00245##
[0811] 33a)
3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol
##STR00246##
[0813] Potassium acetate (2.46 g, 25.02 mmol) was added to
4-bromo-3-methyl-phenol (1.04 g, 5.56 mmol) in
N,N-dimethylformamide (14 mL) at room temperature under argon.
Then, 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane
(2.12 g, 8.34 mmol) was added to the reaction mixture, followed by
the addition of
dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium(II)
dichloromethane adduct (227.0 mg, 278.0 .mu.mol), and the reaction
vessel was evacuated and purged with argon. The mixture was heated
at 80.degree. C. in an oil bath for nineteen hours, then allowed to
cool to room temperature, and concentrated. Water was added,
followed by diethyl ether, and the mixture was filtered through
Celite.RTM.. The filtrate was extracted with diethyl ether and the
organic layer was dried over anhydrous magnesium sulfate, then
filtered and concentrated. The residue was purified by silica gel
chromatography eluting with 1:6 ethyl acetate:hexanes to give 912.4
mg (70%) of
3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol as a
solid. .sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 9.60 (s, 1H),
7.45 (d, J=8 Hz, 1H), 6.56-6.51 (m, 2H), 2.36 (s, 3H), 1.25 (s,
12H). ESI-LCMS m/z 233 (M-H).sup.-.
33b) (4-hydroxy-2-methylphenyl)boronic acid
##STR00247##
[0815] Sodium periodate (2.39 g, 11.15 mmol) was added to
3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol
(870.4 mg, 3.72 mmol) and ammonium acetate (859.8 mg, 11.15 mmol)
in acetone and water (2:1, 36 mL) at room temperature. The mixture
was stirred for sixty-eight hours, then filtered, and concentrated.
Sodium chloride was added to the filtrate, and it was extracted
with ethyl acetate. The organic layer was dried over anhydrous
magnesium sulfate, then filtered and concentrated. The residue was
purified by silica gel chromatography eluting with 1:4 ethyl
acetate:hexanes to give 275.7 mg (49%) of
(4-hydroxy-2-methylphenyl)boronic acid as a solid. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 9.28 (s, 1H), 7.70 (d, J=9 Hz, 1H),
6.56-6.52 (m, 2H), 2.55 (s, 3H). ESI-LCMS m/z 151 (M-H).sup.-.
33c) Ethyl 6-(4-hydroxy-2-methylphenyl)-2-quinolinecarboxylate
##STR00248##
[0817] Palladium(II) acetate (3.2 mg, 14.5 .mu.mol) was added to
ethyl 6-bromo-2-quinolinecarboxylate (81.1 mg, 289.5 .mu.mol),
(4-hydroxy-2-methylphenyl)boronic acid (66.0 mg, 434.2 .mu.mol),
triphenylphosphine (7.6 mg, 29.0 .mu.mol), and potassium phosphate
(215.1 mg, 1.01 mmol). Then, dioxane (2.8 mL) was added to the
mixture, followed by water (289.5 .mu.L), and the reaction mixture
was heated open to the atmosphere at 60.degree. C. in an oil bath
for 18 hours, then allowed to cool to room temperature. Water was
added, followed by ethyl acetate, and the mixture was filtered
through Celite.RTM.. The filtrate was extracted with ethyl acetate,
and the organic layer was dried over anhydrous magnesium sulfate,
then filtered and concentrated. The residue was purified by silica
gel chromatography eluting with 2:3 ethyl acetate:hexanes to give
53.2 mg (60%) of ethyl
6-(4-hydroxy-2-methylphenyl)-2-quinolinecarboxylate as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 9.51 (s, 1H), 8.56 (d,
J=9 Hz, 1H), 8.16 (d, J=9 Hz, 1H), 8.13 (d, J=9 Hz, 1H), 7.98 (s,
1H), 7.82 (d, J=9 Hz, 1H), 7.16 (d, J=8 Hz, 1H), 6.74 (s, 1H), 6.71
(dd, J=8, 2 Hz, 1H), 4.43 (q, J=7 Hz, 2H), 2.23 (s, 3H), 1.39 (t,
J=7 Hz, 3H). ESI-LCMS m/z 308 (M+H).sup.+.
33d) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)-2-methylphenyl]-2-quinolinecarboxylate
##STR00249##
[0819] Cesium carbonate (55.4 mg, 170.0 .mu.mol) was added to ethyl
6-(4-hydroxy-2-methylphenyl)-2-quinolinecarboxylate (47.5 mg, 154.5
.mu.mol) and
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(51.8 mg, 170.0 .mu.mol) in N,N-dimethylformamide (3.1 mL) at room
temperature. The resulting suspension was stirred for eighteen
hours, then water added and the reaction mixture was extracted with
ethyl acetate. The organic layer was dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 3:7 ethyl acetate:hexanes
to give 60.2 mg (68%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-2-methylphenyl]-2-quinolinecarboxylate as an oil. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 8.56 (d, J=9 Hz, 1H), 8.16 (d, J=9 Hz,
1H), 8.13 (d, J=9 Hz, 1H), 7.97 (d, J=2 Hz, 1H), 7.80 (dd, J=9, 2
Hz, 1H), 7.66-7.60 (m, 2H), 7.56 (dd, J=9, 7 Hz, 1H), 7.19 (d, J=8
Hz, 1H), 6.78 (d, J=2 Hz, 1H), 6.73 (dd, J=9, 2 Hz, 1H), 4.86 (s,
2H), 4.43 (q, J=7 Hz, 2H), 3.47 (septet, J=7 Hz, 1H), 2.21 (s, 3H),
1.38 (t, J=7 Hz, 3H), 1.35 (d, J=7 Hz, 6H). ESI-LCMS m/z 575
(M+H).sup.+.
33e)
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)-2-methylphenyl]-2-quinolinecarboxylic acid
##STR00250##
[0821] A 1 N solution of sodium hydroxide (187.7 .mu.L, 187.7
.mu.mol) was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-2-methylphenyl]-2-quinolinecarboxylate (54.0 mg, 93.8 .mu.mol) in
tetrahydrofuran and methanol (1:1, 1.9 mL) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(187.7 .mu.L), water was added, and the reaction mixture was
extracted with ethyl acetate. The organic layer was washed with
brine, then dried over anhydrous magnesium sulfate, filtered and
concentrated to give 50.4 mg (98%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
-2-methylphenyl]-2-quinolinecarboxylic acid as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 13.41 (br s, 1H), 8.54 (d, J=9 Hz,
1H), 8.14 (d, J=9 Hz, 1H), 8.12 (d, J=9 Hz, 1H), 7.96 (s, 1H), 7.79
(dd, J=9, 2 Hz, 1H), 7.66-7.60 (m, 2H), 7.56 (dd, J=9, 7 Hz, 1H),
7.19 (d, J=9 Hz, 1H), 6.78 (d, J=2 Hz, 1H), 6.73 (dd, J=8, 2 Hz,
1H), 4.86 (s, 2H), 3.47 (septet, J=7 Hz, 1H), 2.21 (s, 3H), 1.35
(d, J=7 Hz, 6H). HRMS C.sub.30H.sub.24Cl.sub.2N.sub.2O.sub.4 m/z
547.1191 (M+H).sup.+.sub.Cal; 547.1188 (M+H).sup.-.sub.Obs.
Example 34
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-4-methyl-2-quinolinecarboxylic acid
##STR00251##
[0822] 34a) 1-(5-bromo-2-nitrophenyl)-ethanol
##STR00252##
[0824] Methyl magnesium bromide (7.97 mL of 3.0 M in
tetrahydrofuran, 23.91 mmol) was added dropwise to
5-bromo-2-nitro-benzaldehyde (5.00 g, 21.73 mmol) in
tetrahydrofuran (72 mL) at -78.degree. C. After the addition was
complete, the reaction was stirred for 15 min at -78.degree. C.
Then, the mixture was quenched with water and diethyl ether was
added. The reaction was allowed to warm to room temperature and 10%
citric acid solution was added. The layers were separated and the
organic layer was dried over anhydrous magnesium sulfate, then
filtered and concentrated. The residue was purified by silica gel
chromatography eluting with 1:7 ethyl acetate:hexanes to give 1.68
g (31%) of 1-(5-bromo-2-nitrophenyl)-ethanol as a solid. .sup.1H
NMR (400 MHz, d.sub.6-DMSO): .delta. 7.95 (d, J=2 Hz, 1H), 7.87 (d,
J=9 Hz, 1H), 7.71 (dd, J=9, 2 Hz, 1H), 5.64 (d, J=4 Hz, 1H),
5.16-5.09 (m, 1H), 1.36 (d, J=6 Hz, 3H).
34b) 1-(5-Bromo-2-nitrophenyl)-ethanone
##STR00253##
[0826] Jones Reagent (2.67 M, 2.54 mL, 6.79 mmol) was added
dropwise to 1-(5-bromo-2-nitrophenyl)-ethanol (1.67 g, 6.79 mmol)
in acetone (34 mL) at 0.degree. C., then the reaction mixture was
stirred for 10 minutes. Any excess Jones reagent was quenched by
the addition of iso-propanol, then the reaction mixture was
neutralized with saturated sodium bicarbonate, and the acetone was
removed by evaporation. Water was added and the solution was
extracted with diethyl ether. The organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:9
ethyl acetate:hexanes to give 1.27 g (77%) of
1-(5-bromo-2-nitrophenyl)-ethanone as a solid. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 8.05 (d, J=5 Hz, 1H), 8.04 (s, 1H),
7.96 (dd, J=9, 2 Hz, 1H), 2.57 (s, 3H).
34c) Ethyl 6-bromo-4-methyl-2-quinolinecarboxylate and
1-(2-amino-5-bromophenyl)ethanone
##STR00254##
[0828] A 2.2 M solution of zinc(II) chloride (9.31 mL, 20.49 mmol)
in diethyl ether was added to tin(II) chloride (3.88 g, 20.49 mmol)
and activated 4 .ANG. molecular sieve pellets (1.00 g) in ethanol
(10 mL) under argon. Then, 1-(5-bromo-2-nitrophenyl)-ethanone (1.00
g, 4.10 mmol) and ethyl pyruvate (499.6 mg, 4.30 mmol) in ethanol
(10 mL) were added to the reaction mixture via canula. The mixture
was heated at 70.degree. C. in an oil bath for three hours, then
allowed to cool to room temperature, and carefully quenched with
saturated sodium bicarbonate. Ethyl acetate was added and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:6
ethyl acetate:hexanes to give 39.4 mg (3%) of ethyl
6-bromo-4-methyl-2-quinolinecarboxylate as a solid and 543.1 mg
(62%) of 145-bromo-2-aminophenyl)-ethanone as a solid.
[0829] Ethyl 6-bromo-4-methyl-2-quinolinecarboxylate: .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 8.39 (d, J=2 Hz, 1H), 8.09 (d, J=9
Hz, 1H), 8.02 (s, 1H), 7.97 (dd, J=9, 2 Hz, 1H), 4.41 (q, J=7 Hz,
2H), 2.75 (s, 3H), 1.37 (t, J=7 Hz, 3H). ESI-LCMS m/z 294
(M+H).sup.+. 1-(2-Amino-5-bromophenyl)ethanone: .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 7.82 (d, J=2 Hz, 1H), 7.35 (dd, J=7, 2
Hz, 1H), 7.31 (br s, 2H), 6.73 (d, J=9 Hz, 1H), 2.50 (s, 3H).
ESI-LCMS m/z 213 (M+H).sup.+.
34d)
3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-({[4-(4,4,5,5-tetramethyl--
1,3,2-dioxaborolan-2-yl)phenyl]oxy}methyl)isoxazole
##STR00255##
[0831] Cesium carbonate (1.96 g, 6.03 mmol) was added to
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-phenol (1.26 g,
5.74 mmol) and
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxaz-
ole (1.75 g, 5.74 mmol) in N,N-dimethylformamide (57 mL) at room
temperature. The resulting suspension was heated for sixty-nine
hours at 60.degree. C. The reaction was cooled and concentrated to
remove the N,N-dimethylformamide, then water was added and the
reaction mixture was extracted with diethyl ether. The organic
layer was dried over anhydrous magnesium sulfate, then filtered and
concentrated. The residue was purified by silica gel chromatography
eluting with 1:6 ethyl acetate:hexanes to give 2.24 g (80%) of
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-({[4-(4,4,5,5-tetramethyl-1,3,-
2-dioxaborolan-2-yl)phenyl]oxy}methyl)isoxazole as an solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 7.62-7.59 (m, 2H),
7.55-7.48 (m, 3H), 6.76 (d, J=9 Hz, 2H), 4.82 (s, 2H), 3.44
(septet, J=7 Hz, 1H), 1.31 (d, J=7 Hz, 6H), 1.24 (s, 12H). ESI-LCMS
m/z 488 (M+H).sup.+.
34e) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-4-methyl-2-quinolinecarboxylate
##STR00256##
[0833] Palladium(II) acetate (1.2 mg, 5.4 .mu.mol) was added to
ethyl 6-bromo-4-methyl-2-quinolinecarboxylate (31.5 mg, 107.1
.mu.mol),
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-({[4-(4,4,5,5-tetramethyl-1,3,-
2-dioxaborolan-2-yl)phenyl]oxy}methyl)isoxazole (52.3 mg, 107.1
.mu.mol), triphenylphosphine (2.8 mg, 10.7 .mu.mol), and potassium
phosphate (79.6 mg, 374.8 .mu.mol). Then, dioxane (2.1 mL) was
added to the mixture, followed by water (10.7 .mu.L), and the
reaction mixture was heated open to the atmosphere at 60.degree. C.
in an oil bath for 17 hours, then allowed to cool to room
temperature. Water was added, followed by ethyl acetate, and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate, and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:2
ethyl acetate:hexanes to give 32.1 mg (52%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-methyl-2-quinolinecarboxylate as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 8.25 (d, J=2 Hz, 1H), 8.17 (d, J=9
Hz, 1H), 8.10 (dd, J=9, 2 Hz, 1H), 7.98 (s, 1H), 7.78 (d, J=9 Hz,
2H), 7.65-7.62 (m, 2H), 7.55 (dd, J=9, 7 Hz, 1H), 6.94 (d, J=9 Hz,
2H), 4.89 (s, 2H), 4.41 (q, J=7 Hz, 2H), 3.48 (septet, J=7 Hz, 1H),
2.82 (s, 3H), 1.38 (t, J=7 Hz, 3H), 1.35 (d, J=7 Hz, 6H). ESI-LCMS
m/z 575 (M+H).sup.+.
34f)
6-[4-({[3-(2,6-Dichlorophenyl-5-(1-methylethyl-4-isoxazolyl]methyl}ox-
y)phenyl]-4-methyl-2-quinolinecarboxylic acid
##STR00257##
[0835] A 1 N solution of sodium hydroxide (93.1 .mu.L, 93.1
.mu.mol) was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-methyl-2-quinolinecarboxylate (26.8 mg, 46.6 .mu.mol) in
tetrahydrofuran and methanol (1:1, 931.4 .mu.L) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(93.1 .mu.L), water was added, and the reaction mixture was
extracted with ethyl acetate. The organic layer was washed with
brine, then dried over anhydrous magnesium sulfate, filtered and
concentrated to give 23.1 mg (91%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazoly]met-
hyl}oxy)phenyl]-4-methyl-2-quinolinecarboxylic acid as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 13.29 (br s, 1H), 8.24
(d, J=2 Hz, 1H), 8.16 (d, J=9 Hz, 1H), 8.10 (dd, J=9, 2 Hz, 1H),
7.98 (s, 1H), 7.77 (d, J=9 Hz, 2H), 7.65-7.62 (m, 2H), 7.55 (dd,
J=9, 7 Hz, 1H), 6.94 (d, J=9 Hz, 2H), 4.89 (s, 2H), 3.48 (septet,
J=7 Hz, 1H), 2.81 (s, 3H), 1.35 (d, J=7 Hz, 6H). HRMS
C.sub.3oH.sub.24Cl.sub.2N.sub.2O.sub.4 m/z 547.1191
(M+H).sup.+.sub.Cal; 547.1199 (M+H).sup.+.sub.Obs.
Example 35
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-2-methyl-3-quinolinecarboxylic acid
##STR00258##
[0836] 35a) Ethyl 6-bromo-2-methyl-3-quinolinecarboxylate
##STR00259##
[0838] A 2.2 M solution of zinc(II) chloride (9.88 mL, 21.74 mmol)
in diethyl ether was added to tin(II) chloride (4.12 g, 21.74 mmol)
and activated 4 .ANG. molecular sieve pellets (1.00 g) in ethanol
(10 mL) under argon. Then, 5-bromo-2-nitro-benzaldehyde (1.00 g,
4.35 mmol) and ethyl 3-oxobutanoate (594.0 mg, 4.56 mmol) in
ethanol (12 mL) were added to the reaction mixture via canula. The
mixture was heated at 70.degree. C. in an oil bath for three hours,
then allowed to cool to room temperature, and carefully quenched
with saturated sodium bicarbonate. Ethyl acetate was added and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:6
ethyl acetate:hexanes to give 697.9 mg (55%) of ethyl
6-bromo-2-methyl-3-quinolinecarboxylate as a solid. .sup.lfiNMR
(400 MHz, d.sub.6-DMSO): .delta. 8.84 (s, 1H), 8.42 (d, J=2 Hz,
1H), 7.95 (dd, J=9, 2 Hz, 1H), 7.91 (d, J=9 Hz, 1H), 4.38 (q, J=7
Hz, 2H), 2.84 (s, 3H), 1.37 (t, J=7 Hz, 3H). ESI-LCMS m/z 294
(M+H).sup.+.
35b) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-2-methyl-3-quinolinecarboxylate
##STR00260##
[0840] Palladium(II) acetate (1.2 mg, 5.3 .mu.mol) was added to
ethyl 6-bromo-2-methyl-3-quinolinecarboxylate (31.0 mg, 105.5
.mu.mol),
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-({[4-(4,4,5,5-tetramethyl-1,3,-
2-dioxaborolan-2-yl)phenyl]oxy}methyl)isoxazole (51.5 mg, 105.5
[mol), triphenylphosphine (2.7 mg, 10.5 .mu.mol), and potassium
phosphate (78.3 mg, 369.2 .mu.mol). Then, dioxane (2.1 mL) was
added to the mixture, followed by water (10.5 .mu.L), and the
reaction mixture was heated open to the atmosphere at 60.degree. C.
in an oil bath for 21 hours, then allowed to cool to room
temperature. Water was added, followed by ethyl acetate, and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate, and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 3:7
ethyl acetate:hexanes to give 31.6 mg (52%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-methyl-3-quinolinecarboxylate as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 8.87 (s, 1H), 8.32 (d, J=2 Hz,
1H), 8.11 (dd, J=9, 2 Hz, 1H), 7.99 (d, J=9 Hz, 1H), 7.70 (d, J=9
Hz, 2H), 7.66-7.60 (m, 2H), 7.55 (dd, J=9, 7 Hz, 1H), 6.93 (d, J=9
Hz, 2H), 4.88 (s, 2H), 4.38 (q, J=7 Hz, 2H), 3.48 (septet, J=7 Hz,
1H), 2.85 (s, 3H), 1.38 (t, J=7 Hz, 3H), 1.34 (d, J=7 Hz, 6H).
ESI-LCMS m/z 575 (M+H).sup.+.
35c)
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-methyl-3-quinolinecarboxylic acid
##STR00261##
[0842] A 1 N solution of sodium hydroxide (92.8 .mu.L, 92.8 [mol)
was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-methyl-3-quinolinecarboxylate (26.7 mg, 46.4 .mu.mol) in
tetrahydrofuran and methanol (1:1, 927.9 .mu.L) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(92.8 .mu.L), water was added, and the reaction mixture was
extracted with ethyl acetate. The organic layer was washed with
brine, then dried over anhydrous magnesium sulfate, filtered and
concentrated to give 25.1 mg (99%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]me-
thyl}oxy)phenyl]-2-methyl-3-quinolinecarboxylic acid as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 13.31 (br s, 1H), 8.86
(s, 1H), 8.29 (d, J=2 Hz, 1H), 8.09 (dd, J=9, 2 Hz, 1H), 7.98 (d,
J=9 Hz, 1H), 7.69 (d, J=9 Hz, 2H), 7.66-7.60 (m, 2H), 7.55 (dd,
J=9, 7 Hz, 1H), 6.93 (d, J=9 Hz, 2H), 4.88 (s, 2H), 3.47 (septet,
J=7 Hz, 1H), 2.86 (s, 3H), 1.34 (d, J=7 Hz, 6H). HRMS
C.sub.3oH.sub.24Cl.sub.2N.sub.2O.sub.4 m/z 547.1191
(M+H).sup.+.sub.Cal; 547.1191 (M+H).sup.+.sub.Obs.
Example 36
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-1-isoquinolinecarboxylic acid
##STR00262##
[0843] 36a) N-[2-(3-Bromophenyl)ethyl]benzenesulfonamide
##STR00263##
[0845] Benzenesulfonyl chloride (1.54 mL, 12.00 mmol) was added to
[2-(3-bromophenyl)ethyl]amine (2.00 g, 10.00 mmol) and
N,N-di-iso-propylethylamine (5.22 mL, 29.99 mmol) in
dichloromethane (33 mL) at 0.degree. C. under argon and the
reaction was allowed to warm to room temperature and stirred for 16
hours. The dichloromethane was concentrated under vacuum; diethyl
ether was then added, followed by 10% citric acid. The solution was
extracted with diethyl ether and the organic layer was washed with
saturated sodium bicarbonate, dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 1:4 ethyl acetate:hexanes
to give 3.22 g (95%) of
N-[2-(3-bromophenyl)ethyl]benzenesulfonamide as a solid. .sup.1H
NMR (400 MHz, d.sub.6-DMSO): .delta. 7.77-7.73 (m, 2H), 7.70 (t,
J=6 Hz, 1H), 7.62 (d, J=7 Hz, 1H), 7.57 (t, J=8 Hz, 2H), 7.40-7.34
(m, 2H), 7.22 (t, J=8 Hz, 1H), 7.15 (d, J=8 Hz, 1H), 2.96 (q, J=7
Hz, 2H), 2.67 (t, J=7 Hz, 2H). ESI-LCMS m/z 340 (M+H).sup.+.
36b) Ethyl chloro(methylthio)acetate
##STR00264##
[0847] N-Chlorosuccinimide (5.97 g, 44.71 mmol) was added
portionwise to ethyl (methylthio)acetate (6.00 g, 44.71 mmol) in
carbon tetrachloride (30 mL) at 0.degree. C. under argon and the
reaction was allowed to warm to room temperature and stirred for 17
hours. The reaction mixture was filtered and the filtrate was
concentrated under vacuum. The oil was purified by distillation to
give 3.22 g (43%) of ethyl chloro(methylthio)acetate as an oil.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 6.10 (s, 1H), 4.20 (q,
J=7 Hz, 2H), 2.24 (s, 3H), 1.22 (t, J=7 Hz, 3H).
36c) Ethyl
6-bromo-2-(phenylsulfonyl)-1,2,3,4-tetrahydro-1-isoquinolinecar-
boxylate and Ethyl
8-bromo-2-(phenylsulfonyl)-1,2,3,4-tetrahydro-1-isoquinolinecarboxylate
##STR00265##
[0849] Tin (IV) chloride (6.67 mL of a 1 M solution in
dichloromethane, 6.67 mmol) was added to
N-[2-(3-bromophenyl)ethyl]benzenesulfonamide (1.08 g, 3.17 mmol)
and ethyl chloro(methylthio)acetate (1.07 g, 6.35 mmol) in
1,2-dichloroethane (11 mL) under argon and the reaction was heated
at reflux for 3 hours. After cooling, the mixture was poured into
ice water and extracted with ethyl acetate. The organic layer was
washed with saturated sodium bicarbonate and brine, dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:4
ethyl acetate:hexanes to give 1.07 g (79%) of ethyl
6-bromo-2-(phenylsulfonyl)-1,2,3,4-tetrahydro-1-isoquinolinecarboxylate
and 255.6 mg (19%) of ethyl
8-bromo-2-(phenylsulfonyl)-1,2,3,4-tetrahydro-1-isoquinolinecarboxylate
as oils. Ethyl
6-bromo-2-(phenylsulfonyl)-1,2,3,4-tetrahydro-1-isoquinolinecarboxylate:
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 7.81 (d, J=8 Hz, 2H),
7.64 (t, J=8 Hz, 1H), 7.55 (t, J=8 Hz, 2H), 7.43-7.37 (m, 2H), 7.33
(d, J=8 Hz, 1H), 5.56 (s, 1H), 4.03-3.86 (m, 2H), 3.74-3.56 (m,
2H), 2.84-2.64 (m, 2H), 1.06 (t, J=7 Hz, 3H). ESI-LCMS m/z 424
(M+H).sup.+.
[0850] Ethyl
8-bromo-2-(phenylsulfonyl)-1,2,3,4-tetrahydro-1-isoquinolinecarboxylate:
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 7.76 (d, J=8 Hz, 2H),
7.62 (t, J=8 Hz, 1H), 7.53 (t, J=8 Hz, 2H), 7.48 (d, J=8 Hz, 1H),
7.19-7.12 (m, 2H), 5.73 (s, 1H), 4.06-3.92 (m, 2H), 3.74-3.66 (m,
1H), 3.48-3.40 (m, 1H), 2.80-2.74 (m, 2H), 1.07 (t, J=7 Hz, 3H).
ESI-LCMS m/z 424 (M+H).sup.+.
36d) Ethyl 6-bromo-1-isoquinolinecarboxylate
##STR00266##
[0852] 1,8-Diazabicyclo[5.4.0]undec-7-ene (762.3 .mu.L, 5.10 mmol)
was added to ethyl
6-bromo-2-(phenylsulfonyl)-1,2,3,4-tetrahydro-1-isoquinolinecarboxylate
(1.03 g, 2.43 mmol) in toluene (12 mL) under argon and the reaction
was stirred for 18 hours. Water was added and the mixture was
extracted with ethyl acetate. The organic layer was washed with
saturated sodium bicarbonate, dried over anhydrous magnesium
sulfate, then filtered and concentrated. The residue was purified
by silica gel chromatography eluting with 1:3 ethyl acetate:hexanes
to give 400.6 mg (59%) of ethyl 6-bromo-1-isoquinolinecarboxylate
as a solid. .sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 8.63 (d,
J=6 Hz, 1H), 8.43 (d, J=2 Hz, 1H), 8.41 (d, J=10 Hz, 1H), 8.05 (d,
J=6 Hz, 1H), 7.91 (dd, J=9, 2 Hz, 1H), 4.46 (q, J=7 Hz, 2H), 1.37
(t, J=7 Hz, 3H). ESI-LCMS m/z 280 (M+H).sup.+.
36e) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-1-isoquinolinecarboxylate
##STR00267##
[0854] Palladium(II) acetate (2.5 mg, 11.2 .mu.mol) was added to
ethyl 6-bromo-1-isoquinolinecarboxylate (62.7 mg, 223.8 .mu.mol),
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-({[4-(4,4,5,5-tetramethyl-1,3,-
2-dioxaborolan-2-yl)phenyl]oxy}methyl)isoxazole (109.3 mg, 223.8
.mu.mol), triphenylphosphine (5.9 mg, 22.4 .mu.mol), and potassium
phosphate (166.3 mg, 783.4 .mu.mol). Then, dioxane (2.2 mL) was
added to the mixture, followed by water (22.4 .mu.L), and the
reaction mixture was heated open to the atmosphere at 60.degree. C.
in an oil bath for 67 hours, then allowed to cool to room
temperature. Water was added, followed by ethyl acetate, and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate, and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 2:3
ethyl acetate:hexanes to give 53.6 mg (43%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-1-isoquinolinecarboxylate as an oil. .sup.1H NMR (400 MHz,
d.sub.6-DMSO): .delta. 8.56 (d, J=5 Hz, 1H), 8.47 (d, J=9 Hz, 1H),
8.30 (s, 1H), 8.08-8.03 (m, 2H), 7.75 (d, J=9 Hz, 2H), 7.65-7.60
(m, 2H), 7.55 (dd, J=9, 7 Hz, 1H), 6.95 (d, J=9 Hz, 2H), 4.90 (s,
2H), 4.47 (q, J=7 Hz, 2H), 3.48 (septet, J=7 Hz, 1H), 1.38 (t, J=7
Hz, 3H), 1.34 (d, J=7 Hz, 6H). ESI-LCMS m/z 560 (M+H).sup.+.
36f)
6-[4-({[3-(2,6-Dichlorophenyl-5-(1-methylethyl-4-isoxazolyl]methyl}ox-
y)phenyl]-1-isoquinolinecarboxylic acid
##STR00268##
[0856] A 1 N solution of sodium hydroxide (156.4 .mu.L, 156.4
.mu.mol) was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-1-isoquinolinecarboxylate (43.9 mg, 78.2 .mu.mol) in
tetrahydrofuran and methanol (1:1, 1.6 mL) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(156.4 .mu.L), water was added, and the reaction mixture was
extracted with ethyl acetate. The organic layer was washed with
brine, then dried over anhydrous magnesium sulfate, filtered and
concentrated to give 19.9 mg (48%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-1-isoquinolinecarboxylic acid as a solid. .sup.1H NMR (400
MHz, d.sub.6-DMSO): .delta. 13.64 (br s, 1H), 8.63 (d, J=9 Hz, 1H),
8.55 (d, J=6 Hz, 1H), 8.29 (s, 1H), 8.06 (m, 2H), 7.75 (d, J=9 Hz,
2H), 7.65-7.61 (m, 2H), 7.55 (dd, J=9, 7 Hz, 1H), 6.95 (d, J=9 Hz,
2H), 4.90 (s, 2H), 3.48 (septet, J=7 Hz, 1H), 1.34 (d, J=7 Hz, 6H).
ESI-LCMS m/z 533 (M+H).sup.+.
Example 37
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-4-methyl-2-quinazolinecarboxylic acid
##STR00269##
[0857] 37a) Ethyl [(2-acetyl-4-bromophenyl)amino](oxo)acetate
##STR00270##
[0859] Ethyl chlorooxoacetate (352.7 .mu.L, 3.15 mmol) was added to
1-(2-amino-5-bromophenyl)ethanone (519.8 mg, 2.43 mmol, Example
34c) and pyridine (589.1 .mu.L, 7.28 mmol) in dichloromethane (8
mL) at 0.degree. C. under argon and the reaction was allowed to
warm to room temperature and stirred for 17 hours. Water was added,
and the solution was extracted with diethyl ether. The organic
layer was washed with 10% citric acid, then saturated sodium
bicarbonate, dried over anhydrous magnesium sulfate, filtered, and
concentrated. The residue was purified by silica gel chromatography
eluting with 1:4 ethyl acetate:hexanes to give 648.7 mg (85%) of
ethyl [(2-acetyl-4-bromophenyl)amino](oxo)acetate as a solid.
.sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta. 12.61 (s, 1H), 8.47
(d, J=9 Hz, 1H), 8.26 (d, J=2 Hz, 1H), 7.88 (dd, J=9, 2 Hz, 1H),
4.31 (q, J=7 Hz, 2H), 2.69 (s, 3H), 1.32 (t, J=7 Hz, 3H). ESI-LCMS
m/z 314 (M+H).sup.+.
37b) Ethyl 6-bromo-4-methyl-2-quinazolinecarboxylate
##STR00271##
[0861] Ammonium acetate (400.0 mg, 5.19 mmol) was added to ethyl
[(2-acetyl-4-bromophenyl)amino](oxo)acetate (163.0 mg, 518.9
.mu.mol) in acetic acid (5.2 mL) at room temperature and the
reaction was heated at reflux for 19 hours. The reaction mixture
was concentrated, then water was added, and the solution was
extracted with ethyl acetate. The organic layer was dried over
anhydrous magnesium sulfate, filtered, and concentrated. The
residue was purified by silica gel chromatography eluting with 2:3
ethyl acetate:hexanes to give 75.5 mg (49%) of ethyl
6-bromo-4-methyl-2-quinazolinecarboxylate as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 8.63 (d, J=2 Hz, 1H), 8.21 (dd,
J=9, 2 Hz, 1H), 8.07 (d, J=9 Hz, 1H), 4.41 (q, J=7 Hz, 2H), 2.97
(s, 3H), 1.35 (t, J=7 Hz, 3H). ESI-LCMS m/z 295 (M+H).sup.+.
37c) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-4-methyl-2-quinazolinecarboxylate
##STR00272##
[0863] Palladium(II) acetate (2.7 mg, 12.1 .mu.mol) was added to
ethyl 6-bromo-4-methyl-2-quinazolinecarboxylate (71.1 mg, 240.9
.mu.mol),
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-({[4-(4,4,5,5-tetramethyl-1,3,-
2-dioxaborolan-2-yl)phenyl]oxy}methyl)isoxazole (176.4 mg, 361.4
.mu.mol), triphenylphosphine (6.3 mg, 24.1 .mu.mol), and potassium
phosphate (179.0 mg, 843.2 .mu.mol). Then, dioxane (2.4 mL) was
added to the mixture, followed by water (24.1 .mu.L), and the
reaction mixture was heated open to the atmosphere at 60.degree. C.
in an oil bath for 14 hours, then allowed to cool to room
temperature. Water was added, followed by ethyl acetate, and the
mixture was filtered through Celite.RTM.. The filtrate was
extracted with ethyl acetate, and the organic layer was dried over
anhydrous magnesium sulfate, then filtered and concentrated. The
residue was purified by silica gel chromatography eluting with 1:1
ethyl acetate:hexanes to give 126.2 mg (91%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-methyl-2-quinazolinecarboxylate as a solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 8.44 (d, J=2 Hz, 1H), 8.34 (dd,
J=9, 2 Hz, 1H), 8.15 (d, J=9 Hz, 1H), 7.81 (d, J=9 Hz, 2H),
7.65-7.62 (m, 2H), 7.55 (dd, J=9, 7 Hz, 1H), 6.95 (d, J=9 Hz, 2H),
4.90 (s, 2H), 4.41 (q, J=7 Hz, 2H), 3.48 (septet, J=7 Hz, 1H), 3.03
(s, 3H), 1.36 (t, J=7 Hz, 3H), 1.35 (d, J=7 Hz, 6H). ESI-LCMS m/z
576 (M+H).sup.+.
37d)
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-4-methyl-2-quinazolinecarboxylic acid
##STR00273##
[0865] A 1 N solution of sodium hydroxide (384.4 .mu.L, 384.4
.mu.mol) was added to ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-methyl-2-quinazolinecarboxylate (110.8 mg, 192.2 .mu.mol)
in tetrahydrofuran and methanol (1:1, 1.9 mL) and the mixture was
heated in the microwave at 90.degree. C. for 10 minutes. After
cooling, the mixture was neutralized with 1 N hydrochloric acid
(384.4 .mu.L), water added, and the reaction mixture was extracted
with ethyl acetate. The organic layer was washed with brine, then
dried over anhydrous magnesium sulfate, filtered and concentrated
to give 104.0 mg (99%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-4-methyl-2-quinazolinecarboxylic acid as a solid. .sup.1H
NMR (400 MHz, d.sub.6-DMSO): .delta. 13.47 (br s, 1H), 8.43 (d, J=2
Hz, 1H), 8.33 (dd, J=9, 2 Hz, 1H), 8.12 (d, J=9 Hz, 1H), 7.81 (d,
J=9 Hz, 2H), 7.65-7.62 (m, 2H), 7.55 (dd, J=9 7 Hz, 1H), 6.95 (d,
J=9 Hz, 2H), 4.90 (s, 2H), 3.48 (septet, J=7 Hz, 1H), 3.03 (s, 3H),
1.35 (d, J=7 Hz, 6H). ESI-LCMS m/z 548 (M+H).sup.+.
Example 38
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio)-
phenyl]-2-quinolinecarboxylic acid
##STR00274##
[0866] 38a)
[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio)p-
henyl]boronic acid
##STR00275##
[0868] A mixture of
4-(chloromethyl)-3-(2,6-dichlorophenyl)-5-(1-methylethyl)isoxazole
(152 mg, 0.5 mmol), cesium carbonate (489 mg, 1.5 mmol) and
4-mercaptophenyl boronic acid (75 mg, 0.5 mmol) in
dimethylformamide (5 mL) was allowed to stir for 24 hours in a
60.degree. C. oil bath. The mixture was diluted with ethyl acetate
and washed with water. The organic layer was washed with brine,
dried with magnesium sulfate, filtered and concentrated to yield
277 mg of crude
[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio)p-
henyl]boronic acid and dimethylformamide. .sup.1H NMR (400 MHz,
d.sub.6-DMSO): .delta. 7.63-7.59 (m, 4H), 7.55-7.51 (m, 1H), 7.17
(d, J=8 Hz, 2H), 3.86 (s, 2H), 3.12 (septet, J=7 Hz, 1H), 1.13 (d,
J=7 Hz, 6H).
38b) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}thio)phenyl]-2-quinolinecarboxylate
##STR00276##
[0870] A mixture of crude
[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio)p-
henyl]boronic acid (76 mg, 0.18 mmol), ethyl
6-bromo-2-quinolinecarboxylate (42 mg, 0.15 mmol),
tetrakis(triphenylphosphine)palladium(0) (7 mg, 0.0006 mmol) in
1,2-dimethoxyethane (0.8 mL), and 2 M sodium carbonate (0.65 mL)
was heated in a 70.degree. C. oil bath for 1.5 hours. The solution
was partitioned between ethyl acetate and water. The organic layer
was washed with brine, dried with magnesium sulfate filtered and
concentrated. The residue was purified by silica gel chromatography
(hexane to 2:5 ethyl acetate:hexanes gradient elution) to give 20
mg (22% as 0.4 ethyl acetate) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio-
)phenyl]-2-quinolinecarboxylate. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 8.38-8.32 (m, 2H), 8.21 (d, J=9 Hz, 1H), 7.98 (s, 1H), 7.96
(d, J=2 Hz, 1H), 7.59 (d, J=8 Hz, 2H), 7.43-7.32 (m, 5H), 4.57 (q,
J=7 Hz, 2H), 3.82 (s, 2H), 3.07 (septet, J=7 Hz, 1H), 1.50 (t, J=7
Hz, 3H), 1.27 (d, J=7 Hz, 6H).
38c)
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
thio)phenyl]-2-quinolinecarboxylic acid
##STR00277##
[0872] A solution of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio-
)phenyl]-2-quinolinecarboxylate (20 mg, 0.033 mmol) in
tetrahydrofuran (0.33 mL) was placed in a microwave reaction vial
followed by ethanol (0.17 mL) and 1 N sodium hydroxide (0.050 mL).
The tube was sealed then heated to 100.degree. C. for ten minutes.
The reaction mixture was then concentrated and the residue
partitioned between tetrahydrofuran and brine. 1 N hydrochloric
acid (0.050 mL) was added. The organic layer was separated and
concentrated to yield 17 mg (94%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}thio-
)phenyl]-2-quinolinecarboxylic acid as an orange solid. .sup.1H NMR
(400 MHz, d.sub.6-DMSO): .delta. 8.57 (d, J=9 Hz, 1H), 8.35 (s,
1H), 8.20-8.10 (m, 3H), 7.76 (d, J=8 Hz, 2H), 7.63 (d, J=7 Hz, 2H),
7.58-7.7.54 (m, 1H), 7.42 (d, J=8 Hz, 2H), 3.95 (s, 2H), 3.24
(septet, J=7 Hz, 1H), 1.16 (d, J=7 Hz, 6H). HRMS (ESI)
C.sub.29H.sub.22Cl.sub.2N.sub.2O.sub.3S calculated: 549.0806
[M+H].sup.+, found: 549.0813 [M+H].sup.+.
Example 39
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)p-
henyl]-2-quinoxalinecarboxylic acid
##STR00278##
[0873] 39a) Ethyl 6-bromo-2-quinoxalinecarboxylate
##STR00279##
[0875] To a stirred solution of 4-bromo-o-phenylenediamine (3.1 g,
16.6 mmol) in 1-methyl-2-pyrrolidinone (150 mL) was added dropwise
ethyl bromopyruvate at room temperature under nitrogen. After 20
hours, the reaction mixture was partitioned between water and
diethyl ether. The organic phase was separated and the aqueous
phase was extracted with diethyl ether twice more. The organic
extracts were combined, washed twice with water, dried over
magnesium sulfate, filtered, and the filtrate was concentrated to
give an oil. The crude product was partially purified by flash
chromatography over silica with dichloromethane to give a tan
solid. The impure product was purified on a Chiralpak AS-H column
with 95% carbon dioxide at 140 bar at 35.degree. C. and 2 mL/min
with 5% co-solvent (methanol:chloroform (80:20) to give 0.614 g
(13%) of ethyl 6-bromo-2-quinoxalinecarboxylate as a tan solid.
.sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 9.52 (s, 1H), 8.37 (d,
J=2 Hz, 1H), 8.16 (d, J=9 Hz, 1H), 7.93 (dd, J=9, 2 Hz, 1H), 4.59
(quartet, J=7 Hz, 2H), 1.50 (t, J=7 Hz, 3H). ESI-LCMS m/z 281
(M+H).sup.+.
39b) Ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]m-
ethyl}oxy)phenyl]-2-quinoxalinecarboxylate
##STR00280##
[0877] Ethyl 6-bromo-2-quinoxalinecarboxylate (0.108 g, 0.384
mmol),
3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-({[4-(4,4,5,5-tetramethyl-1,3,-
2-dioxaborolan-2-yl)phenyl]oxy}methyl)isoxazole (0.27 g, 0.553
mmol), potassium phosphate (0.256 g, 1.21 mmol), palladium (II)
acetate (0.0088 g, 0.039 mmol), triphenylphosphine (0.013 g, 0.050
mmol), 1,4-dioxane (8 mL) and water (0.035 mL) were combined and
heated at 60.degree. C. with stirring overnight. After 20 hours,
the reaction mixture was allowed to stand at room temperature. The
reaction mixture was partitioned between water and ethyl acetate.
The aqueous phase was separated and extracted a second time with
ethyl acetate. The organic extracts were combined, washed with
brine, dried over magnesium sulfate, filtered, and the filtrate was
concentrated to give an oil. The crude product was purified by
flash chromatography over silica with hexanes:ethyl acetate (100:0
to 50:50) to give a yellow solid. The solid was dissolved in
dichloromethane and the solution was concentrated to give 0.12 g
(56%) of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinoxalinecarboxylate as a yellow amorphous solid.
.sup.1H NMR (CDCl.sub.3, 400 MHz): .delta. 9.52 (s, 1H), 8.31 (d,
J=9 Hz, 1H), 8.27 (d, J=2 Hz, 1H), 8.06 (dd, J=9, 2 Hz, 1H), 7.65
(d, J=9 Hz, 2H), 7.41 (m, 2H), 7.32 (dd, J=9, 7 Hz, 1H), 6.92 (d,
J=9 Hz, 2H), 4.80 (s, 2H), 4.60 (quartet, J=7 Hz, 2H), 3.36
(septet, J=7 Hz, 1H), 1.51 (t, J=7 Hz, 3H), 1.44 (d, J=7 Hz, 6H).
ESI-LCMS m/z 562 (M+H).sup.+.
39c)
6-[4-({[3-(2,6-Dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}-
oxy)phenyl]-2-quinoxalinecarboxylic acid
##STR00281##
[0879] To a stirred solution of ethyl
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinoxalinecarboxylate (0.112 g, 0.20 mmol) in ethanol (5
mL) and tetrahydrofuran (5 mL) was added 1 N sodium hydroxide (0.21
mL, 0.21 mmol). The reaction mixture was stirred at room
temperature under nitrogen for 3.5 hours. The solvent was removed
in vacuo and water (5 mL) was added to the resulting solid. The pH
of the aqueous mixture was adjusted to .about.3 (litmus paper) with
1 N HCl. The acidic aqueous mixture was extracted with ethyl
acetate. The aqueous phase was separated and extracted a second
time with ethyl acetate. The aqueous phase was separated and found
to contain product according to thin layer chromatography. The pH
of the aqueous phase was adjusted to .about.2 (litmus paper) with 1
N HCl. The acidic aqueous phase was combined with the
aforementioned ethyl acetate extracts. The organic phase was
separated, washed with water followed by brine, dried over
magnesium sulfate, filtered, and the filtrate was concentrated to
give a yellow solid which was dried under high vacuum at room
temperature to give 0.095 g (89%) of
6-[4-({[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methyl}oxy)-
phenyl]-2-quinoxalinecarboxylic acid. .sup.1H NMR (d.sub.6-DMSO,
400 MHz): .delta. 13.89 (br s, 1H), 9.40 (s, 1H), 8.32 (s, 1H),
8.24 (m, 2H), 7.81 (d, J=9 Hz, 2H), 7.62 (m, 2H), 7.53 (dd, J=9, 7
Hz, 1H), 6.95 (d, J=9 Hz, 2H), 4.89 (s, 2H), 3.47 (septet, J=7 Hz,
1H), 1.33 (d, J=7 Hz, 6H). ESI-LCMS m/z 532 (M-H).sup.-.
Biological Example 40
FXR Cofactor Binding Assay
[0880] Determination of a ligand mediated cofactor peptide
interaction to quantify ligand binding to the nuclear receptor
Farnesoid X Receptor (FXR). The method measures the ability of
putative ligands to modulate the interaction between the purified
bacterial expressed FXR.alpha. ligand binding domain (LBD) and a
synthetic biotinylated peptide based on residues 676-700 of steroid
receptor coactivator-1 (SRC-1) (LXXLL-containing domain-2 where L
is the amino acid leucine and X indicates any other amino acid
(LCD2), 676-700). The sequence of the SRC-1 peptide used is as
published in Iannone, M. A., et al., 2001 Cytometry 44:326-337
where the N-terminus was biotinylated (B) and the C-terminus was
amidated. Detection of the associated complex was measured by time
resolved fluorescence (TRF). The purified LBD of FXR was labeled
with biotin then mixed with stoichiometric amounts of
allophycocyanin (APC) labeled streptavidin (Molecular Probes). The
biotinylated peptide was then mixed with a 1/2 stoichiometric
amount of europium labeled streptavidin (Wallac Inc). Each was then
blocked with a 5 fold excess of biotin and allowed to equilibrate
for 15 min. Equimolar amounts of receptor and peptide were mixed
together and were allowed to equilibrate for at least 30 min prior
to the addition to either a variable or constant concentrations of
the sample for which the affinity is to be determined. After
equilibration, the time-resolved fluorescent signal was quantitated
using a fluorescent plate reader. The affinity of the test compound
was estimated from a plot of fluorescence versus concentration of
test compound added.
[0881] A basal level of FXR: peptide formation is observed in the
absence of added ligand. Ligands that promote the complex formation
induce a concentration-dependent increase in time-resolved
fluorescent signal. Compounds which bind equally well to both
monomeric FXR and to the FXR: peptide complex would be expected to
give no change in signal, whereas ligands which bind preferentially
to the monomeric receptor would be expected to induce a
concentration-dependent decrease in the observed signal.
[0882] Methods & Materials
[0883] Advance Preparation: Human Farnesoid X Receptor .alpha.
Ligand Binding Domain
[0884] Human FXR.alpha. Ligand Binding Domain (FXR.alpha. LBD) was
expressed in E. coli strain BL21 (DE3) as an amino-terminal
polyhistidine tagged fusion protein. Expression was under the
control of an isopropyl-13-D-thiogalactopyranoside (IPTG) inducible
T7 promoter. DNA encoding this recombinant protein is subcloned
into the pRSET-A expression vector (Invitrogen). The coding
sequence of Human FXR.alpha. LBD was derived from Genbank accession
number U 68233 (amino acids 237 to 472).
[0885] Ten-liter fermentation batches were grown in Rich PO.sub.4
media with 0.1 mg/mL Ampicillin at 25.degree. C. for 12 hours,
cooled to 9.degree. C. and held at that temperature for 36 hours to
a density of OD.sub.600=14. At this cell density, 0.25 mM IPTG is
added and induction proceeded for 24 hours at 9.degree. C., to a
final OD.sub.600=16. Cells are harvested by centrifugation (20
minutes, 3500.times.gravity, 4.degree. C.), and concentrated cell
slurries were stored in phosphate buffered saline (PBS) at
-8.degree. C.
[0886] Purification of Receptor Ligand Binding Domain
[0887] Routinely, 30-40 g cell paste (equivalent to 2-3 liters of
the fermentation batch) was resuspended in 200-250 mL Tris buffered
saline (TBS), pH 7.2 (25 mM Tris-hydroxymethylamino methane (Tris),
150 mM sodium chloride). Cells were lysed by passing 3 times
through a French Press and cell debris was removed by
centrifugation (30 minutes, 20,000.times.gravity, 4.degree. C.).
The cleared supernatant was filtered through course pre-filters,
and TBS, pH 7.2, 500 mM imidazole was added to obtain a final
imidazole concentration of 50 mM. This lysate was loaded onto a
column (6.times.8 cm) packed with Sepharose [Ni.sup.||
charged]Chelation resin (Pharmacia) and pre-equilibrated with TBS
pH 7.2/50 mM imidazole. After washing to baseline absorbance with
equilibration buffer, the column was washed with one column volume
of TBS pH 7.2 containing 90 mM imidazole. FXR.alpha. LBD was eluted
directly with 365 mM imidazole. Column fractions were pooled and
dialyzed against TBS, pH 7.2, containing 0.5 mM EDTA and 5 mM DTT.
The dialyzed protein sample was concentrated using Centri-prep 10 K
(Amicon) and subjected to size exclusion, using a column
(3.times.90 cm) packed with Sepharose S-75 resin (Pharmacia)
pre-equilibrated with TBS, pH 7.2, containing 0.5 mM ethylene
diamine tetraacetic acid (EDTA) and 5 mM dithiothreitol (DTT).
[0888] Biotinylation of FXR
[0889] Purified FXR.alpha. LBD was desalted/buffer exchanged using
PD-10 gel filtration columns into PBS [100 mM Na.sub.2PO.sub.4, pH
7.2, 150 mM NaCl]. FXR.alpha. LBD was diluted to approximately 60
.mu.M in PBS and five-fold molar excess of NHS-LC-Biotin (Pierce)
is added in a minimal volume of PBS. This solution was incubated
with gentle mixing for 30 minutes at room temperature. The
biotinylation modification reaction was stopped by the addition of
2000.times.molar excess of Tris-HCl, pH 8. The modified FXR.alpha.
LBD was dialyzed against 4 buffer changes, each of at least 50
volumes, PBS containing 5 mM DTT, 2 mM EDTA and 2% sucrose. The
biotinylated FXR.alpha. LBD was then subjected to mass
spectrometric analysis to reveal the extent of modification by the
biotinylation reagent. In general, approximately 95% of the protein
had at least a single site of biotinylation; and the overall extent
of biotinylation followed a normal distribution of multiple sites,
ranging from zero to four.
[0890] Preparation of Streptavidin-(Europium
Chelate)-SRC1:Streptavdin-(APC)-FXR Complex
[0891] Biotinylated SRC-1 (LCD2, 676-700) peptide and a 1/2
stoichiometric amount of streptavidin-conjugated europium chelate
was incubated in assay buffer containing 10 mM DTT for at least 30
minutes. A second solution of stoichiometric amounts of
biotinylated FXR and streptavidin-conjugated APC was incubated in
assay buffer containing 10 mM DTT for at least 30 minutes. Each
solution was then blocked with a 5 fold molar excess of biotin and
allowed to equilibrate for at least 30 min. The labeled receptor
and cofactor were mixed and again allowed to equilibrate for at
least 30 min, added to the compound plate, utilizing e.g., a
Titertek Multidrop 384.
[0892] Materials:
[0893] Assay Buffer: 50 mM 3-(N-morpholino)propanesulfonic acid
(MOPS) pH 7.5, 50 mM NaF, 50 .mu.M
3-[(3-cholamidopropyl)-demethylammonio]-1-propanesulfonate (CHAPS),
0.1 mg/ml Fraction 5 fatty acid free bovine serum albumin (BSA), 1
mM ethylenediaminetetraacetic acid (EDTA). Solid DTT is added to
the assay buffer to a final concentration of 10 mM just before use
in the assay. BSA, fatty acid free
[0894] DTT
[0895] NaF
[0896] Europium labeled Streptavidin: (Wallac CR28-100)
[0897] 384 well Plates
[0898] Methods:
[0899] Experimental Details:
[0900] Test compounds and controls were serial diluted in DMSO and
0.1 .mu.L at the desired concentration were added to a 384 well
plate.
[0901] To each well to be assayed a previously prepared solution of
FXR-APC and Europium labeled SRC1 was added to 0.1 .mu.L of test
compound and controls for a final assay volume of 10 .mu.L.
[0902] The plates were incubated for at least 1 hour at room
temperature and the fluorescent signal determined in a Fluorescence
Reader in a time resolved mode utilizing e.g., a Wallac Viewlux
Imager or Wallac Victor Multilabel counter.
[0903] Data Reduction:
[0904] For each concentration of test compound, the results of each
test well was expressed as % of control, C, calculated according to
eq. 1.
C = 100 * F sample - F basal F std - F basal ( 1 ) ##EQU00001##
[0905] where F.sub.sample is the signal observed in a particular
sample well, F.sub.total is i the signal observed in the presence
of control inhibitor and F.sub.basal is the count rate observed in
the presence of no ligand. The values used for F.sub.std and
F.sub.basal are averages of the corresponding control wells
included on every plate. The results are reported in Table 1 below.
In Table 1, +indicates a pEC.sub.50 of 5-5.99; ++ indicates a
pEC.sub.50 6-6.99 and +++ indicates a pEC.sub.50 greater than
7.
TABLE-US-00004 TABLE 1 Example Activity (pEC.sub.50) 1g ++ 1h ++ 1i
++ 2 ++ 3 ++ 4 ++ 5 ++ 6 ++ 7 ++ 8 ++ 9 ++ 10 ++ 11 ++ 12 ++ 13 ++
14 ++ 15 ++ 16 ++ 17 ++ 18 ++ 19 ++ 20 ++ 21 ++ 22 ++ 23 ++ 24 + 25
++ 26 ++ 27 + 28 ++ 29 ++ 30 ++ 31 ++ 32 ++ 33 ++ 34 +++ 35 ++ 36
++ 37 ++ 38 ++ 39 ++
Example 41
The Effects of FXR Agonists on Diet Induced Obese Mice
[0906] Methods: Male 20 to 25 g C57BL mice (Charles River,
Indianapolis, Ind.) were housed at 72.degree. F. and 50% relative
humidity with a 12 h light and dark cycle and fed with standard
rodent chow (Purina 5001, Harlan Teklad, Indianapolis, Ind.) or a
high-fat diet (TD93075, Harlan Teklad, Indianapolis, Ind.) for
eight weeks. After three weeks, mice on high-fat diet were
randomized to vehicle or treatment groups based on fasting glucose
and body weight. Starting from the fifth week, mice were given
either vehicle or the compound in Example 1 (10, 30, and 100 mg/kg)
or the compound in Example 5 (10, 30, 100 mg/kg) twice a day
orally. Mice on the standard rodent chow were also given vehicle as
a control. Body composition was measured using the quantitative
magnetic resonance (QMR) method before and at the end of compound
treatment. At the end of the study (fourth week of compound
treatment), blood samples were taken from inferior vena cava and
tissue samples were collected for further analysis. Serum chemistry
levels were measured using the Instrumentation Laboratory Ilab600TM
clinical chemistry analyzer (Instrumentation Laboratory, Boston,
Mass.).
[0907] Results: The compound in Example 1 and the compound in
Example 5 decreased body fat mass, serum glucose, insulin,
cholesterol, triglyceride, NEFA, and glycerol in high-fat diet fed
obese mice.
* * * * *