U.S. patent application number 17/703891 was filed with the patent office on 2022-07-07 for ghrelin o-acyltransferase inhibitors.
This patent application is currently assigned to Glaxosmithkline Intellectual Property Development Limited. The applicant listed for this patent is Glaxosmithkline Intellectual Property Development Limited. Invention is credited to Anish Bandyopadhyay, Mui Cheung, Hilary Schenck Eidam, Hemant Joshi, Dai-Shi Su.
Application Number | 20220213076 17/703891 |
Document ID | / |
Family ID | |
Filed Date | 2022-07-07 |
United States Patent
Application |
20220213076 |
Kind Code |
A1 |
Bandyopadhyay; Anish ; et
al. |
July 7, 2022 |
GHRELIN O-ACYLTRANSFERASE INHIBITORS
Abstract
This invention relates to novel compounds according to Formula
(I) which are inhibitors of ghrelin O-acyltransferase (GOAT), to
pharmaceutical compositions containing them, to processes for their
preparation, and to their use in therapy for the treatment of
metabolic disorders (e.g. Prader-Willi syndrome, metabolic
syndrome, insulin resistance, impaired glucose tolerance,
prediabetes, diabetes mellitus (e.g., type II diabetes mellitus),
dysglycemia (e.g., hyperglycemia), obesity (e.g., obesity caused by
Prader-Willi syndrome), increased adiposity, poor glycemic control,
hyperphagia, impaired satiety, dyslipidemia (e.g., atherogenic
dyslipidemia), hepatic steatosis (e.g., non-alcoholic fatty liver
disease (e.g., non-alcoholic steatohepatitis))), psychiatric
disorders (e.g., eating disorders (e.g., bulimia nervosa, binge
eating disorder, night-time eating syndrome), substance related
disorders (e.g., addiction disorders (e.g., alcohol, smoking,
overeating, or use of illicit drugs))), as well as disorders
related to or complications of metabolic or psychiatric disorders
(e.g., cardiovascular diseases (e.g., diabetic heart disease (e.g.,
diabetic cardiomyopathy), heart failure, or hypertension), ischemia
(e.g., myocardial ischemia, cerebral ischemia, ischemic stroke), or
BMI-related cancers (e.g., pancreatic cancer, gallbladder cancer,
esophageal cancer, colorectal cancer, breast cancer etc.).
##STR00001##
Inventors: |
Bandyopadhyay; Anish;
(Hyderabad, IN) ; Cheung; Mui; (King Of Prussia,
PA) ; Eidam; Hilary Schenck; (Stevenage, GB) ;
Joshi; Hemant; (Hyderabad, IN) ; Su; Dai-Shi;
(Collegeville, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Glaxosmithkline Intellectual Property Development Limited |
Brentford |
|
GB |
|
|
Assignee: |
Glaxosmithkline Intellectual
Property Development Limited
Brentford
GB
|
Appl. No.: |
17/703891 |
Filed: |
March 24, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16967262 |
Aug 4, 2020 |
11312709 |
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PCT/EP2019/052770 |
Feb 5, 2019 |
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17703891 |
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International
Class: |
C07D 409/12 20060101
C07D409/12; C07D 491/048 20060101 C07D491/048 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 5, 2018 |
IN |
201811004277 |
Claims
1-30. (canceled)
31. A method of preparing a compound of Formula (I): ##STR00135##
or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is
hydrogen, halogen, cyano, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, or --C(.dbd.O)NH.sub.2; X is CH.sub.2
or O; R.sup.2 is halogen; and R.sup.3 is hydrogen or halogen;
comprising hydrolyzing a compound of Formula (IV): ##STR00136## or
a salt thereof, wherein: R.sup.1 is hydrogen, halogen, cyano,
(C.sub.1-C.sub.4)alkyl, halo(C.sub.1-C.sub.4)alkyl, or
--C(.dbd.O)NH.sub.2; X is CH.sub.2 or O; R.sup.2 is halogen;
R.sup.3 is hydrogen or halogen; and R.sup.4 is
(C.sub.1-C.sub.4)alkyl; in the presence of water and a base.
32. The method of claim 31, wherein the compound of Formula (I) is
of the Formula (II): ##STR00137## or a pharmaceutically acceptable
salt thereof.
33. The method of claim 31, wherein the compound of Formula (I) is
of the Formula (III): ##STR00138## or a pharmaceutically acceptable
salt thereof.
34. The method of claim 31, wherein the base is a hydroxide
base.
35. The method of claim 34, wherein the hydroxide base is lithium
hydroxide.
36. The method of claim 34, wherein the hydroxide base is sodium
hydroxide.
37. The method of claim 31, further comprising a solvent.
38. The method of claim 37, wherein the solvent comprises an
alcohol.
39. The method of claim 38, wherein the solvent further comprises
water.
40. The method of claim 38, wherein the alcohol is methanol.
41. The method of claim 31, wherein the compound of Formula (IV) is
of the formula: ##STR00139## or a salt thereof.
42. The method of claim 31, wherein the compound of Formula (I) is
of the formula: ##STR00140## or a pharmaceutically acceptable salt
thereof.
43. A method of preparing a compound of the formula: ##STR00141##
or a salt thereof, wherein: R.sup.1 is hydrogen, halogen, cyano,
(C.sub.1-C.sub.4)alkyl, halo(C.sub.1-C.sub.4)alkyl, or
--C(.dbd.O)NH.sub.2; X is CH.sub.2 or O; R.sup.2 is halogen;
R.sup.3 is hydrogen or halogen; and R.sup.4 is
(C.sub.1-C.sub.4)alkyl; comprising contacting a first compound of
the formula: ##STR00142## or a salt thereof, wherein: R.sup.1 is
hydrogen, halogen, cyano, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, or --C(.dbd.O)NH.sub.2; X is CH.sub.2
or O; and R.sup.5 is --OH, or --Cl; with a second compound of the
formula: ##STR00143## or a salt thereof.
44. The method of claim 43, wherein the first compound is of the
formula: ##STR00144## or a salt thereof.
45. The method of claim 43, wherein the first compound is of the
formula: ##STR00145## or a salt thereof.
46. The method of claim 43, wherein the first compound is of the
formula: ##STR00146## or a salt thereof.
47. The method of claim 43, wherein the second compound is of the
formula: ##STR00147## or a salt thereof.
48. The method of claim 43, further comprising contacting the
compounds in the presence of a base.
49. The method of claim 48, wherein the base is potassium
carbonate.
50. The method of claim 43, further comprising contacting the
compounds in the presence of a phosphine.
Description
RELATED APPLICATIONS
[0001] The present application claims priority under 35 U.S.C.
.sctn. 119(a) to Indian patent application, Application Number
201811004277, filed Feb. 5, 2018, the entire contents of which are
incorporated herein by reference.
FIELD OF THE INVENTION
[0002] This invention relates to compounds which inhibit ghrelin
O-acyltransferase (GOAT) and thus are useful for reducing appetite
and adiposity, as well as improving energy balance and glycemic
control.
BACKGROUND OF THE INVENTION
[0003] Ghrelin is a 28-amino acid gastric hormone produced
primarily in the fundus of the stomach. Two forms of the hormone
are found in circulation: unacylated ghrelin and acylated ghrelin.
The lone enzyme known to perform this post-translational acylation
on serine 3 of ghrelin is ghrelin O-acyltransferase (GOAT). There
is no other known function of GOAT. Only acyl ghrelin is capable of
interacting with its receptor, growth hormone secretagogue receptor
1 (GHSR1). Binding of acyl ghrelin to GHSR1 in the brain stimulates
orexigenic activity and adiposity and reduces energy expenditure.
When acyl ghrelin was administered to humans, appetite and food
intake were increased (covered in Cummings, Physiology &
Behavior 2006, 89, 71-84). Binding of acyl ghrelin to GHSR1 in
pancreatic islet cells modulates insulin release. Acute
administration of acyl ghrelin to humans led to significant
reductions in plasma insulin and increased glucose levels (Broglio,
J. Clin. Endocrinol. Metab. 2001, 86, 5083-5086).
[0004] Levels of acylated ghrelin increase in anticipation of a
meal and decrease post-prandially, leading acyl ghrelin to dubbed
the "hunger hormone." If increased levels of acyl ghrelin stimulate
adiposity and adversely impact glycemic control, which could
contribute to the development of the metabolic syndrome, then
decreasing the amount of acyl ghrelin in circulation should do the
opposite: reduce appetite and adiposity, improve energy balance,
and benefit glycemic control, potentially ameliorating the
metabolic syndrome. Inhibition of GOAT decreases acyl ghrelin
production. Indeed, as reviewed by Ariyasu and Akamizu (Endocrine
Journal 2015, 62(11), 953-963) mice with ghrelin, GHSR1, or GOAT
knocked out demonstrate decreased food intake on a high fat diet
and increased insulin secretion. When wild type mice were
administered a peptide-based GOAT inhibitor, they demonstrated
reduced weight gain and improved glucose tolerance (Barnett et al.,
Science 2010, 330 (6011), 1689-1692). In addition, it was recently
reported that ghrelin deletion is protective against age-associated
hepatic steatosis, suggesting a role for GOAT inhibition in the
treatment of nonalcoholic steatohepatitis (NASH) (Guillory et al.,
Aging Cell 2017 published ahead of print 10.1111/acel.12688).
[0005] Thus, there is strong evidence to suggest that inhibition of
GOAT decreases appetite and adiposity and improves glycemic
control. Accordingly, compounds that inhibit GOAT activity would be
useful for the treatment of obesity.
SUMMARY OF THE INVENTION
[0006] The present invention relates to compounds according to
Formula (I) or pharmaceutically acceptable salts thereof:
##STR00002##
wherein:
[0007] R.sup.1 is hydrogen, halogen, cyano, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, or --C(.dbd.O)NH.sub.2;
[0008] X is CH.sub.2 or O;
[0009] R.sup.2 is halogen; and
[0010] R.sup.3 is hydrogen or halogen.
[0011] Exemplary compounds of Formula (I) include, but are not
limited to:
##STR00003## ##STR00004##
pharmaceutically acceptable salts thereof.
[0012] Another aspect of this invention relates to a method of
treating obesity. In particular, this invention relates to a method
of treating obesity caused by Prader-Willi syndrome. Prader-Willi
syndrome is a well-known genetic cause of obesity and is found in
people of both sexes and in all races worldwide, particularly in
children. Patients suffering from Prader-Willi syndrome experience
hyperphagia and typically have trouble controlling their weight.
Many complications of Prader-Willi syndrome are due to obesity.
[0013] Another aspect of this invention relates to a method of
treating metabolic disorders (e.g. Prader-Willi syndrome, metabolic
syndrome, insulin resistance, impaired glucose tolerance,
prediabetes, diabetes mellitus (e.g., type II diabetes mellitus),
dysglycemia (e.g., hyperglycemia), obesity (e.g., obesity caused by
Prader-Willi syndrome), increased adiposity, poor glycemic control,
hyperphagia, impaired satiety, dyslipidemia (e.g., atherogenic
dyslipidemia), hepatic steatosis (e.g., non-alcoholic fatty liver
disease (e.g., non-alcoholic steatohepatitis))), psychiatric
disorders (e.g., eating disorders (e.g., bulimia nervosa, binge
eating disorder, night-time eating syndrome), substance related
disorders (e.g., addiction disorders (e.g., alcohol, smoking,
overeating, or use of illicit drugs))), as well as disorders
related to or complications of metabolic or psychiatric disorders
(e.g., cardiovascular diseases (e.g., diabetic heart disease (e.g.,
diabetic cardiomyopathy), heart failure, or hypertension), ischemia
(e.g., myocardial ischemia, cerebral ischemia, ischemic stroke), or
BMI-related cancers (e.g., pancreatic cancer, gallbladder cancer,
esophageal cancer, colorectal cancer, breast cancer etc.).
[0014] Another aspect of the invention relates to pharmaceutical
preparations comprising compounds of Formula (I) and
pharmaceutically acceptable excipients.
[0015] In another aspect, there is provided the use of a compound
of Formula (I) or a pharmaceutically acceptable salt or solvate
thereof, in the manufacture of a medicament for use in the
treatment of a disorder mediated by GOAT, such as obesity. In
another aspect, there is provided the use of a compound of Formula
(I) or a pharmaceutically acceptable salt or solvate thereof, in
the manufacture of a medicament for use in the treatment of a
disorder mediated by GOAT, such as metabolic disorders (e.g.
Prader-Willi syndrome, metabolic syndrome, insulin resistance,
impaired glucose tolerance, prediabetes, diabetes mellitus (e.g.,
type II diabetes mellitus), dysglycemia (e.g., hyperglycemia),
obesity (e.g., obesity caused by Prader-Willi syndrome), increased
adiposity, poor glycemic control, hyperphagia, impaired satiety,
dyslipidemia (e.g., atherogenic dyslipidemia), hepatic steatosis
(e.g., non-alcoholic fatty liver disease (e.g., non-alcoholic
steatohepatitis))), psychiatric disorders (e.g., eating disorders
(e.g., bulimia nervosa, binge eating disorder, night-time eating
syndrome), substance related disorders (e.g., addiction disorders
(e.g., alcohol, smoking, overeating, or use of illicit drugs))), as
well as disorders related to or complications of metabolic or
psychiatric disorders (e.g., cardiovascular diseases (e.g.,
diabetic heart disease (e.g., diabetic cardiomyopathy), heart
failure, or hypertension), ischemia (e.g., myocardial ischemia,
cerebral ischemia, ischemic stroke), or BMI-related cancers (e.g.,
pancreatic cancer, gallbladder cancer, esophageal cancer,
colorectal cancer, breast cancer etc.).
[0016] In another aspect, the invention provides a compound of
Formula (I) or a pharmaceutically acceptable salt thereof for use
in therapy.
[0017] In another aspect, there is provided a compound of Formula
(I) or a pharmaceutically acceptable salt thereof for use in the
treatment of a disorder mediated by GOAT.
[0018] In another aspect, there is provided a compound of Formula
(I) or a pharmaceutically acceptable salt thereof for use in the
treatment of obesity.
[0019] In another aspect, there is provided a compound of Formula
(I) or a pharmaceutically acceptable salt thereof for use in the
treatment of metabolic disorders (e.g. Prader-Willi syndrome,
metabolic syndrome, insulin resistance, impaired glucose tolerance,
prediabetes, diabetes mellitus (e.g., type II diabetes mellitus),
dysglycemia (e.g., hyperglycemia), obesity (e.g., obesity caused by
Prader-Willi syndrome), increased adiposity, poor glycemic control,
hyperphagia, impaired satiety, dyslipidemia (e.g., atherogenic
dyslipidemia), hepatic steatosis (e.g., non-alcoholic fatty liver
disease (e.g., non-alcoholic steatohepatitis))), psychiatric
disorders (e.g., eating disorders (e.g., bulimia nervosa, binge
eating disorder, night-time eating syndrome), substance related
disorders (e.g., addiction disorders (e.g., alcohol, smoking,
overeating, or use of illicit drugs))), as well as disorders
related to or complications of metabolic or psychiatric disorders
(e.g., cardiovascular diseases (e.g., diabetic heart disease (e.g.,
diabetic cardiomyopathy), heart failure, or hypertension), ischemia
(e.g., myocardial ischemia, cerebral ischemia, ischemic stroke), or
BMI-related cancers (e.g., pancreatic cancer, gallbladder cancer,
esophageal cancer, colorectal cancer, breast cancer etc.).
[0020] In another aspect, there is provided a compound of Formula
(I) or a pharmaceutically acceptable salt thereof for use in the
treatment of Prader-Willi syndrome.
[0021] In another aspect, provided herein are methods of
co-administering the presently invented compounds of Formula (I)
with other active ingredients.
[0022] In another aspect, there is provided a combination of a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof and at least one anti-adiposity agent or anti-adiposity
therapy for use in the treatment of a disorder mediated by
GOAT.
[0023] In another aspect, there is provided a combination of a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof and at least one lifestyle modification (e.g., a
reduced-calorie diet and/or exercise), weight loss agent (e.g.,
orlistat, lorcaserin, liraglutide, phentermine/topimarate, or
naltrexone/bupropion), hormone therapy (e.g., testosterone,
estrogen, progesterone, or human growth hormone), selective
serotonin reuptake inhibitors (SSRIs), or anti-diabetic therapy
(e.g., insulin, miglitol, acarbose, metformin, exenatide,
pramlintide) for use in the treatment of a disorder mediated by
GOAT.
[0024] In another aspect, there is provided a combination of a
compound of Formula (I) or a pharmaceutically acceptable salt
thereof and at least one anti-adiposity agent or anti-adiposity
therapy for use in the treatment of obesity. In another aspect,
there is provided a combination of a compound of Formula (I) or a
pharmaceutically acceptable salt thereof and at least one lifestyle
modification (e.g., a reduced-calorie diet and/or exercise), weight
loss agent (e.g., orlistat, lorcaserin, liraglutide,
phentermine/topimarate, or naltrexone/bupropion), hormone therapy
(e.g., testosterone, estrogen, progesterone, or human growth
hormone), selective serotonin reuptake inhibitors (SSRIs), and/or
anti-diabetic therapy (e.g., insulin, miglitol, acarbose,
metformin, exenatide, pramlintide) for use in the treatment of
metabolic disorders (e.g. Prader-Willi syndrome, metabolic
syndrome, insulin resistance, impaired glucose tolerance,
prediabetes, diabetes mellitus (e.g., type II diabetes mellitus),
dysglycemia (e.g., hyperglycemia), obesity (e.g., obesity caused by
Prader-Willi syndrome), increased adiposity, poor glycemic control,
hyperphagia, impaired satiety, dyslipidemia (e.g., atherogenic
dyslipidemia), hepatic steatosis (e.g., non-alcoholic fatty liver
disease (e.g., non-alcoholic steatohepatitis))), psychiatric
disorders (e.g., eating disorders (e.g., bulimia nervosa, binge
eating disorder, night-time eating syndrome), substance related
disorders (e.g., addiction disorders (e.g., alcohol, smoking,
overeating, or use of illicit drugs))), as well as disorders
related to or complications of metabolic or psychiatric disorders
(e.g., cardiovascular diseases (e.g., diabetic heart disease (e.g.,
diabetic cardiomyopathy), heart failure, or hypertension), ischemia
(e.g., myocardial ischemia, cerebral ischemia, ischemic stroke), or
BMI-related cancers (e.g., pancreatic cancer, gallbladder cancer,
esophageal cancer, colorectal cancer, breast cancer etc.).
[0025] In another aspect, the present disclosure provides
pharmaceutical compositions or preparations including a compound
described herein, and optionally a pharmaceutically acceptable
excipient. In certain embodiments, the pharmaceutical compositions
described herein include a therapeutically or prophylactically
effective amount of a compound described herein. The pharmaceutical
composition or preparation may be useful for treating and/or
preventing a disease (e.g., metabolic disorders (e.g. Prader-Willi
syndrome, metabolic syndrome, insulin resistance, impaired glucose
tolerance, prediabetes, diabetes mellitus (e.g., type II diabetes
mellitus), dysglycemia (e.g., hyperglycemia), obesity (e.g.,
obesity caused by Prader-Willi syndrome), increased adiposity, poor
glycemic control, hyperphagia, impaired satiety, dyslipidemia
(e.g., atherogenic dyslipidemia), hepatic steatosis (e.g.,
non-alcoholic fatty liver disease (e.g., non-alcoholic
steatohepatitis))), psychiatric disorders (e.g., eating disorders
(e.g., bulimia nervosa, binge eating disorder, night-time eating
syndrome), substance related disorders (e.g., addiction disorders
(e.g., alcohol, smoking, overeating, or use of illicit drugs))), as
well as disorders related to or complications of metabolic or
psychiatric disorders (e.g., cardiovascular diseases (e.g.,
diabetic heart disease (e.g., diabetic cardiomyopathy), heart
failure, or hypertension), ischemia (e.g., myocardial ischemia,
cerebral ischemia, ischemic stroke), or BMI-related cancers (e.g.,
pancreatic cancer, gallbladder cancer, esophageal cancer,
colorectal cancer, breast cancer etc.) in a subject in need
thereof. The pharmaceutical composition or preparation may be
useful for inhibiting the activity of GOAT in a subject, biological
sample, tissue, or cell.
[0026] In another aspect, the present disclosure provides
pharmaceutical compositions or preparations including a compound
described herein, and optionally a pharmaceutically acceptable
excipient. In certain embodiments, the pharmaceutical compositions
described herein include a therapeutically or prophylactically
effective amount of a compound described herein. The pharmaceutical
composition or preparation may be useful for treating metabolic
disorders (e.g. Prader-Willi syndrome, metabolic syndrome, insulin
resistance, impaired glucose tolerance, prediabetes, diabetes
mellitus (e.g., type II diabetes mellitus), dysglycemia (e.g.,
hyperglycemia), obesity (e.g., obesity caused by Prader-Willi
syndrome), increased adiposity, poor glycemic control, hyperphagia,
impaired satiety, dyslipidemia (e.g., atherogenic dyslipidemia),
hepatic steatosis (e.g., non-alcoholic fatty liver disease (e.g.,
non-alcoholic steatohepatitis))), psychiatric disorders (e.g.,
eating disorders (e.g., bulimia nervosa, binge eating disorder,
night-time eating syndrome), substance related disorders (e.g.,
addiction disorders (e.g., alcohol, smoking, overeating, or use of
illicit drugs))), as well as disorders related to or complications
of metabolic or psychiatric disorders (e.g., cardiovascular
diseases (e.g., diabetic heart disease (e.g., diabetic
cardiomyopathy), heart failure, or hypertension), ischemia (e.g.,
myocardial ischemia, cerebral ischemia, ischemic stroke), or
BMI-related cancers (e.g., pancreatic cancer, gallbladder cancer,
esophageal cancer, colorectal cancer, breast cancer etc.) in a
subject in need thereof, or inhibiting the activity of GOAT in a
biological sample, tissue, or cell.
[0027] In another aspect, described herein are methods for treating
and/or preventing a disease (e.g., metabolic disorders (e.g.
Prader-Willi syndrome, metabolic syndrome, insulin resistance,
impaired glucose tolerance, prediabetes, diabetes mellitus (e.g.,
type II diabetes mellitus), dysglycemia (e.g., hyperglycemia),
obesity (e.g., obesity caused by Prader-Willi syndrome), increased
adiposity, poor glycemic control, hyperphagia, impaired satiety,
dyslipidemia (e.g., atherogenic dyslipidemia), hepatic steatosis
(e.g., non-alcoholic fatty liver disease (e.g., non-alcoholic
steatohepatitis))), psychiatric disorders (e.g., eating disorders
(e.g., bulimia nervosa, binge eating disorder, night-time eating
syndrome), substance related disorders (e.g., addiction disorders
(e.g., alcohol, smoking, overeating, or use of illicit drugs))), as
well as disorders related to or complications of metabolic or
psychiatric disorders (e.g., cardiovascular diseases (e.g.,
diabetic heart disease (e.g., diabetic cardiomyopathy), heart
failure, or hypertension), ischemia (e.g., myocardial ischemia,
cerebral ischemia, ischemic stroke), or BMI-related cancers (e.g.,
pancreatic cancer, gallbladder cancer, esophageal cancer,
colorectal cancer, breast cancer etc.) in a subject, biological
sample, tissue, or cell.
[0028] Another aspect relates to methods of inhibiting the activity
of GOAT using a compound described herein in a biological sample
(e.g., cell, or tissue). In another aspect, described herein are
methods of inhibiting the activity of GOAT using a compound
described herein in a subject.
[0029] In another aspect, the present disclosure provides compounds
of Formula (I), and pharmaceutically acceptable salts thereof, for
use in the treatment of a disease (e.g., metabolic disorders (e.g.
Prader-Willi syndrome, metabolic syndrome, insulin resistance,
impaired glucose tolerance, prediabetes, diabetes mellitus (e.g.,
type II diabetes mellitus), dysglycemia (e.g., hyperglycemia),
obesity (e.g., obesity caused by Prader-Willi syndrome), increased
adiposity, poor glycemic control, hyperphagia, impaired satiety,
dyslipidemia (e.g., atherogenic dyslipidemia), hepatic steatosis
(e.g., non-alcoholic fatty liver disease (e.g., non-alcoholic
steatohepatitis))), psychiatric disorders (e.g., eating disorders
(e.g., bulimia nervosa, binge eating disorder, night-time eating
syndrome), substance related disorders (e.g., addiction disorders
(e.g., alcohol, smoking, overeating, or use of illicit drugs))), as
well as disorders related to or complications of metabolic or
psychiatric disorders (e.g., cardiovascular diseases (e.g.,
diabetic heart disease (e.g., diabetic cardiomyopathy), heart
failure, or hypertension), ischemia (e.g., myocardial ischemia,
cerebral ischemia, ischemic stroke), or BMI-related cancers (e.g.,
pancreatic cancer, gallbladder cancer, esophageal cancer,
colorectal cancer, breast cancer etc.) in a subject, biological
sample, tissue, or cell.
[0030] Another aspect of the present disclosure relates to kits
comprising a container with a compound, or pharmaceutical
composition or preparation thereof, as described herein. The kits
described herein may include a single dose or multiple doses of the
compound or pharmaceutical composition or preparation. The kits may
be useful in a method of the disclosure. In certain embodiments,
the kit further includes instructions for using the compound or
pharmaceutical composition or preparation. A kit described herein
may also include information (e.g., prescribing information) as
required by a regulatory agency, such as the U.S. Food and Drug
Administration (FDA).
[0031] The details of one or more embodiments of the invention are
set forth herein. Other features, objects, and advantages of the
invention will be apparent from the Detailed Description, Examples,
Figures, and Claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0032] FIG. 1 shows an X-ray powder diffraction pattern of the
compound of Example 1a.
[0033] FIG. 2 shows a differential scanning calorimetry trace of
the compound of Example 1a and a thermogravimetric analysis trace
of the compound of Example 1a.
[0034] FIG. 3 shows the dose response of Example 1a on fasting
induced acyl ghrelin levels.
[0035] FIG. 4 shows the effect of Example 1a on fasting-induced
acyl ghrelin levels in male SD rats.
[0036] FIG. 5 shows the effect of Example 1a on fasting-induced
des-acyl ghrelin levels in male SD rats.
[0037] FIG. 6 shows acyl ghrelin reduction after a single 10 mg/kg
dose in Cynomolgus monkeys.
[0038] FIG. 7 shows the effect of Example 1a and Rimonabant on
plasma acyl ghrelin levels in high fat high carbohydrate fed male
C57BL/6 mice.
[0039] FIG. 8 shows the effect of Example 1a and Rimonabant on
plasma des-acyl ghrelin levels in high fat high carbohydrate fed
male C57BL/6 mice.
DETAILED DESCRIPTION OF THE INVENTION
[0040] The present invention provides compounds that inhibit GOAT,
and pharmaceutical compositions/preparations thereof, for the
treatment of a disease in a subject. The present invention further
provides methods of using the compounds described herein, e.g., as
biological probes to study the inhibition of GOAT or ghrelin
activity, and as therapeutics, e.g., in the treatment of diseases
associated with GOAT activity. In certain embodiments, the diseases
include, but are not limited to, metabolic disorders (e.g.
Prader-Willi syndrome, metabolic syndrome, insulin resistance,
impaired glucose tolerance, prediabetes, diabetes mellitus (e.g.,
type II diabetes mellitus), dysglycemia (e.g., hyperglycemia),
obesity (e.g., obesity caused by Prader-Willi syndrome), increased
adiposity, poor glycemic control, hyperphagia, impaired satiety,
dyslipidemia (e.g., atherogenic dyslipidemia), hepatic steatosis
(e.g., non-alcoholic fatty liver disease (e.g., non-alcoholic
steatohepatitis))), psychiatric disorders (e.g., eating disorders
(e.g., bulimia nervosa, binge eating disorder, night-time eating
syndrome), substance related disorders (e.g., addiction disorders
(e.g., alcohol, smoking, overeating, or use of illicit drugs))), as
well as disorders related to or complications of metabolic or
psychiatric disorders (e.g., cardiovascular diseases (e.g.,
diabetic heart disease (e.g., diabetic cardiomyopathy), heart
failure, or hypertension), ischemia (e.g., myocardial ischemia,
cerebral ischemia, ischemic stroke), or BMI-related cancers (e.g.,
pancreatic cancer, gallbladder cancer, esophageal cancer,
colorectal cancer, breast cancer etc.) in a subject, biological
sample, tissue or cell.
[0041] This invention relates to compounds of the Formula (I) as
defined above, or pharmaceutically acceptable salts thereof.
Formula (I) contains the substituent R.sup.1. In certain
embodiments, R.sup.1 is H. In certain embodiments, R.sup.1 is
halogen. In certain embodiments, R.sup.1 is Cl. In certain
embodiments, R.sup.1 is --CN. In certain embodiments, R.sup.1 is
--(C.sub.1-C.sub.4)alkyl. In certain embodiments, R.sup.1 is -Me.
In certain embodiments, R.sup.1 is -Et. In certain embodiments,
R.sup.1 is -Pr. In certain embodiments, R.sup.1 is
-halo(C.sub.1-C.sub.4)alkyl. In certain embodiments, R.sup.1 is
--CF.sub.3. In certain embodiments, R.sup.1 is --C(.dbd.O)NH.sub.2.
In certain embodiments, R.sup.1 is hydrogen, halogen, cyano,
(C.sub.1-C.sub.4)alkyl, halo(C.sub.1-C.sub.4)alkyl,
--C(.dbd.O)N(R.sup.a).sub.2, wherein R.sup.a is hydrogen,
substituted or unsubstituted C.sub.1-C.sub.6 alkyl, or a nitrogen
protecting group. In certain embodiments, R.sup.1 is
--C(.dbd.O)N(R.sup.a).sub.2. In certain embodiments, R.sup.a is
hydrogen. In certain embodiments, R.sup.a is substituted or
unsubstituted C.sub.1-C.sub.6 alkyl. In certain embodiments,
R.sup.a is -Me. In certain embodiments, R.sup.a is -Et. In certain
embodiments, R.sup.a is a nitrogen protecting group.
[0042] Formula (I) contains the substituent X. In certain
embodiments, X is CH.sub.2. In certain embodiments, X is O.
[0043] Formula (I) contains the substituent R.sup.2. In certain
embodiments, R.sup.2 is halogen. In certain embodiments, R.sup.2 is
--Cl.
[0044] Formula (I) contains the substituent R.sup.3. In certain
embodiments, R.sup.3 is H. In certain embodiments, R.sup.3 is
halogen. In certain embodiments, R.sup.3 is F. In certain
embodiments, R.sup.3 is Cl.
[0045] In certain embodiments, X is CH.sub.2, and R.sup.1 is
methyl. In certain embodiments, R.sup.1 is Me, R.sup.2 is Cl, and
R.sup.3 is H. In certain embodiments, X is CH.sub.2, R.sup.1 is
methyl, R.sup.2 is Cl, and R.sup.3 is H.
[0046] Exemplary compounds of Formula (I) include, but are not
limited to:
TABLE-US-00001 Exam- ple Structure 1 ##STR00005## 2 ##STR00006## 3
##STR00007## 4 ##STR00008## 5 ##STR00009## 6 ##STR00010## 7
##STR00011## 8 ##STR00012## 9 ##STR00013##
and pharmaceutically acceptable salts thereof.
[0047] In one embodiment, this invention relates to compounds of
Formula (II) represented by Formula (II):
##STR00014##
wherein:
[0048] R.sup.1 is hydrogen, halogen, cyano, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, or --C(.dbd.O)NH.sub.2;
[0049] X is CH.sub.2 or O;
[0050] R.sup.2 is halogen; and
[0051] R.sup.3 is hydrogen or halogen.
[0052] In certain embodiments, this invention relates to compounds
of Formula (II),
wherein:
[0053] R.sup.1 is hydrogen, halogen, cyano, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, or --C(.dbd.O)N(R.sup.a).sub.2, wherein
R.sup.a is hydrogen, substituted or unsubstituted C.sub.1-C.sub.6
alkyl, or a nitrogen protecting group;
[0054] X is CH.sub.2 or O;
[0055] R.sup.2 is halogen; and
[0056] R.sup.3 is hydrogen or halogen.
[0057] Formula (II) contains the substituent R.sup.1. In certain
embodiments, R.sup.1 is H. In certain embodiments, R.sup.1 is
halogen. In certain embodiments, R.sup.1 is Cl. In certain
embodiments, R.sup.1 is --CN. In certain embodiments, R.sup.1 is
--(C.sub.1-C.sub.4)alkyl. In certain embodiments, R.sup.1 is -Me.
In certain embodiments, R.sup.1 is -halo(C.sub.1-C.sub.4)alkyl. In
certain embodiments, R.sup.1 is --CF.sub.3. In certain embodiments,
R.sup.1 is --C(.dbd.O)NH.sub.2. In certain embodiments, R.sup.1 is
--C(.dbd.O)N(R.sup.a).sub.2. In certain embodiments, R.sup.a is
hydrogen. In certain embodiments, R.sup.a is substituted or
unsubstituted C.sub.1-C.sub.6 alkyl. In certain embodiments,
R.sup.a is -Me. In certain embodiments, R.sup.a is -Et. In certain
embodiments, R.sup.a is a nitrogen protecting group.
[0058] Formula (II) contains the substituent X. In certain
embodiments, X is CH.sub.2. In certain embodiments, X is O.
[0059] Formula (II) contains the substituent R.sup.2. In certain
embodiments, R.sup.2 is halogen. In certain embodiments, R.sup.2 is
--Cl.
[0060] Formula (II) contains the substituent R.sup.3. In certain
embodiments, R.sup.3 is H. In certain embodiments, R.sup.3 is
halogen. In certain embodiments, R.sup.3 is F. In certain
embodiments, R.sup.3 is Cl.
[0061] In certain embodiments, X is CH.sub.2 and R.sup.1 is methyl.
In certain embodiments, R.sup.1 is Me, R.sup.2 is Cl, and R.sup.3
is H. In certain embodiments, X is CH.sub.2, R.sup.1 is methyl,
R.sup.2 is Cl and R.sup.3 is H.
[0062] Exemplary compounds of Formula (II) include, but are not
limited to:
TABLE-US-00002 Exam- ple Structure 1a ##STR00015## 2a ##STR00016##
3a ##STR00017## 4a ##STR00018## 5a ##STR00019## 6a ##STR00020## 7a
##STR00021## 8a ##STR00022## 9a ##STR00023##
and pharmaceutically acceptable salts thereof.
[0063] In certain embodiments, Formula (II) is of the formula:
##STR00024##
In certain embodiments, Formula (II) is of the formula:
##STR00025##
or a pharmaceutically acceptable salt thereof.
[0064] In another embodiment, this invention relates to compounds
of Formula (I) represented by Formula (III):
##STR00026##
wherein
[0065] R.sup.1 is hydrogen, halogen, cyano, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, or --C(.dbd.O)NH.sub.2;
[0066] X is CH.sub.2 or O;
[0067] R.sup.2 is halogen; and
[0068] R.sup.3 is hydrogen or halogen.
In another embodiment, this invention relates to compounds of
Formula (III), wherein:
[0069] R.sup.1 is hydrogen, halogen, cyano, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, or --C(.dbd.O)N(R.sup.a).sub.2, wherein
R.sup.a is hydrogen, substituted or unsubstituted C.sub.1-C.sub.6
alkyl, or a nitrogen protecting group;
[0070] X is CH.sub.2 or O;
[0071] R.sup.2 is halogen; and
[0072] R.sup.3 is hydrogen or halogen.
[0073] Formula (III) contains the substituent R.sup.1. In certain
embodiments, R.sup.1 is H. In certain embodiments, R.sup.1 is
halogen. In certain embodiments, R.sup.1 is Cl. In certain
embodiments, R.sup.1 is --CN. In certain embodiments, R.sup.1 is
--(C.sub.1-C.sub.4)alkyl. In certain embodiments, R.sup.1 is -Me.
In certain embodiments, R.sup.1 is -halo(C.sub.1-C.sub.4)alkyl. In
certain embodiments, R.sup.1 is --CF.sub.3. In certain embodiments,
R.sup.1 is --C(.dbd.O)NH.sub.2. In certain embodiments, R.sup.1 is
--C(.dbd.O)N(R.sup.a).sub.2. In certain embodiments, R.sup.a is
hydrogen. In certain embodiments, R.sup.a is substituted or
unsubstituted C.sub.1-C.sub.6 alkyl. In certain embodiments,
R.sup.a is -Me. In certain embodiments, R.sup.a is -Et. In certain
embodiments, R.sup.a is a nitrogen protecting group.
[0074] Formula (III) contains the substituent X. In certain
embodiments, X is CH.sub.2. In certain embodiments, X is O.
[0075] Formula (III) contains the substituent R.sup.2. In certain
embodiments, R.sup.2 is halogen. In certain embodiments, R.sup.2 is
--Cl.
[0076] Formula (III) contains the substituent R.sup.3. In certain
embodiments, R.sup.3 is H. In certain embodiments, R.sup.3 is
halogen. In certain embodiments, R.sup.3 is F. In certain
embodiments, R.sup.3 is Cl.
[0077] In certain embodiments, X is CH.sub.2 and R.sup.1 is methyl.
In certain embodiments, R.sup.1 is Me, R.sup.2 is Cl, and R.sup.3
is H. In certain embodiments, X is CH.sub.2, R.sup.1 is methyl,
R.sup.2 is Cl and R.sup.3 is H.
[0078] Exemplary compounds of Formula (III) include, but are not
limited to:
TABLE-US-00003 Exam- ple Structure 1b ##STR00027## 2b ##STR00028##
3b ##STR00029## 4b ##STR00030## 5b ##STR00031## 6b ##STR00032## 7b
##STR00033## 8b ##STR00034## 9b ##STR00035##
and pharmaceutically acceptable salts thereof.
[0079] Specific compounds of this invention include: [0080]
2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]thio-
phen-3-yl)acetic acid; [0081]
(R)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid; [0082]
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid; [0083]
2-(4-chloro-6-((2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)ben-
zo[b]thiophen-3-yl)acetic acid; [0084]
(R)-2-(4-chloro-6-((2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetic acid; [0085]
(S)-2-(4-chloro-6-((2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetic acid; [0086]
2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetic acid; [0087]
(R)-2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl-
)oxy)benzo[b]thiophen-3-yl)acetic acid; [0088]
(S)-2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl-
)oxy)benzo[b]thiophen-3-yl)acetic acid; [0089]
2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyrid-
in-5-yl)oxy)benzo[b]thiophen-3-yl)acetic acid; [0090]
(R)-2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]p-
yridin-5-yl)oxy)benzo[b]thiophen-3-yl)acetic acid; [0091]
(S)-2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]p-
yridin-5-yl)oxy)benzo[b]thiophen-3-yl)acetic acid; [0092]
2-(4-chloro-7-fluoro-6-((6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]pyridi-
n-3-yl)oxy)benzo[b]thiophen-3-yl)acetic acid; [0093]
(R)-2-(4-chloro-7-fluoro-6-((6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]py-
ridin-3-yl)oxy)benzo[b]thiophen-3-yl)acetic acid; [0094]
(S)-2-(4-chloro-7-fluoro-6-((6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]py-
ridin-3-yl)oxy)benzo[b]thiophen-3-yl)acetic acid; [0095]
2-(4-chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thiophen-3-
-yl)acetic acid; [0096]
(R)-2-(4-chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thioph-
en-3-yl)acetic acid; [0097]
(S)-2-(4-chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thioph-
en-3-yl)acetic acid; [0098]
2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thioph-
en-3-yl)acetic acid; [0099]
(R)-2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]th-
iophen-3-yl)acetic acid; [0100]
(S)-2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]th-
iophen-3-yl)acetic acid; [0101]
2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benz-
o[b]thiophen-3-yl)acetic acid; [0102]
(R)-2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-
benzo[b]thiophen-3-yl)acetic acid; [0103]
(S)-2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-
benzo[b]thiophen-3-yl)acetic acid; [0104]
2-(6-((2-carbamoyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-4-chloro-
benzo[b]thiophen-3-yl)acetic acid; [0105]
(R)-2-(6-((2-carbamoyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-4-ch-
lorobenzo[b]thiophen-3-yl)acetic acid; and [0106]
(S)-2-(6-((2-carbamoyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-4-ch-
lorobenzo[b]thiophen-3-yl)acetic acid; [0107] and pharmaceutically
acceptable salts thereof.
[0108] Typically, but not absolutely, the salts of the present
invention are pharmaceutically acceptable salts. Salts of the
disclosed compounds containing a basic amine or other basic
functional group may be prepared by any suitable method known in
the art, including treatment of the free base with an inorganic
acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid,
nitric acid, phosphoric acid, and the like, or with an organic
acid, such as acetic acid, trifluoroacetic acid, maleic acid,
succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic
acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid,
such as glucuronic acid or galacturonic acid, alpha-hydroxy acid,
such as citric acid or tartaric acid, amino acid, such as aspartic
acid or glutamic acid, aromatic acid, such as benzoic acid or
cinnamic acid, sulfonic acid, such as p-toluenesulfonic acid,
methanesulfonic acid, ethanesulfonic acid or the like. Examples of
pharmaceutically acceptable salts include sulfates, pyrosulfates,
bisulfates, sulfites, bisulfites, phosphates, chlorides, bromides,
iodides, acetates, propionates, decanoates, caprylates, acrylates,
formates, isobutyrates, caproates, heptanoates, propiolates,
oxalates, malonates succinates, suberates, sebacates, fumarates,
maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates,
chlorobenzoates, methylbenzoates, dinitrobenzoates,
hydroxybenzoates, methoxybenzoates, phthalates, phenylacetates,
phenylpropionates, phenylbutrates, citrates, lactates,
.gamma.-hydroxybutyrates, glycolates, tartrates mandelates, and
sulfonates, such as xylenesulfonates, methanesulfonates,
propanesulfonates, naphthalene-1-sulfonates and
naphthalene-2-sulfonates.
[0109] Salts of the disclosed compounds containing a carboxylic
acid or other acidic functional group can be prepared by reacting
with a suitable base. Such a pharmaceutically acceptable salt may
be made with a base which affords a pharmaceutically acceptable
cation, which includes alkali metal salts (especially sodium and
potassium), alkaline earth metal salts (especially calcium and
magnesium), aluminum salts and ammonium salts, as well as salts
made from physiologically acceptable organic bases such as
trimethylamine, triethylamine, morpholine, pyridine, piperidine,
picoline, dicyclohexylamine, N,N'-dibenzylethylenediamine,
2-hydroxyethylamine, bis-(2-hydroxyethyl)amine,
tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine,
dehydroabietylamine, N,N'-bisdehydroabietylamine, glucamine,
N-methylglucamine, collidine, quinine, quinoline, and basic amino
acid such as lysine and arginine.
[0110] Other salts, which are not pharmaceutically acceptable, may
be useful in the preparation, isolation, or storage of the
compounds of this invention, and these should be considered to form
a further aspect of the invention. These salts, such as oxalic or
trifluoroacetate, while not in themselves pharmaceutically
acceptable, may be useful in the preparation of salts useful as
intermediates in obtaining the compounds of the invention and their
pharmaceutically acceptable salts.
[0111] The compound of Formula (I) may exist in a crystalline or
noncrystalline form, or as a mixture thereof. The skilled artisan
will appreciate that pharmaceutically acceptable solvates may be
formed for crystalline or non-crystalline compounds. In crystalline
solvates, solvent molecules are incorporated into the crystalline
lattice during crystallization. Solvates may involve non-aqueous
solvents such as, but not limited to, ethanol, isopropanol, DMSO,
acetic acid, ethanolamine, or ethyl acetate, or they may involve
water as the solvent that is incorporated into the crystalline
lattice. Solvates wherein water is the solvent incorporated into
the crystalline lattice are typically referred to as "hydrates."
Hydrates include stoichiometric hydrates as well as compositions
containing variable amounts of water. The invention includes all
such solvates.
[0112] The skilled artisan will further appreciate that the
compounds of the invention that exist in crystalline form,
including the various solvates thereof, may exhibit polymorphism
(i.e. the capacity to occur in different crystalline structures).
These different crystalline forms are typically known as
"polymorphs." The invention includes all such polymorphs.
Polymorphs have the same chemical composition but differ in
packing, geometrical arrangement, and other descriptive properties
of the crystalline solid state. Polymorphs, therefore, may have
different physical properties such as shape, density, hardness,
deformability, stability, and dissolution properties. Polymorphs
typically exhibit different melting points, IR spectra, and X-ray
powder diffraction patterns, which may be used for identification.
The skilled artisan will appreciate that different polymorphs may
be produced, for example, by changing or adjusting the reaction
conditions or reagents, used in making the compound. For example,
changes in temperature, pressure, or solvent may result in
polymorphs. In addition, one polymorph may spontaneously convert to
another polymorph under certain conditions.
[0113] The present invention is further directed to crystalline
forms of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid.
[0114] In some embodiments, a crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern comprising at least nine diffraction
angles, when measured using Cu K.sub..alpha. radiation, selected
from a group consisting of about 5.9, 13.6, 14.0, 14.3, 21.9, 22.5,
23.1, 23.3, 24.1, 24.5, 24.7, 25.7, 26.1, 26.6, and 27.4 degrees
2.theta.. In another embodiment, the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern comprising at least eight diffraction
angles or at least seven diffraction angles or at least six
diffraction angles or at least five diffraction angles or at least
four diffraction angles, when measured using Cu K.sub..alpha.
radiation, selected from a group consisting of about 5.9, 13.6,
14.0, 14.3, 21.9, 22.5, 23.1, 23.3, 24.1, 24.5, 24.7, 25.7, 26.1,
26.6, and 27.4 degrees 2.theta.. In another embodiment, the
crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern comprising at least three diffraction
angles, when measured using Cu K.sub..alpha. radiation, selected
from a group consisting of about 5.9, 13.6, 14.0, 14.3, 21.9, 22.5,
23.1, 23.3, 24.1, 24.5, 24.7, 25.7, 26.1, 26.6, and 27.4 degrees
2.theta..
[0115] In another embodiment, the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern comprising diffraction angles, when
measured using Cu K.sub..alpha. radiation, of about 5.9, 13.6,
14.0, 14.3, 23.3, 24.5, and 27.4 degrees 2.theta..
[0116] In yet another embodiment, the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern substantially in accordance with FIG.
1.
[0117] In further embodiments, the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by a differential
scanning calorimetry trace substantially in accordance with FIG. 2
and/or a thermogravimetric analysis trace substantially in
accordance with FIG. 2.
[0118] In still further embodiments, as a person having ordinary
skill in the art will understand,
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by any combination of
the analytical data characterizing the aforementioned embodiments.
For example, in one embodiment, the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern substantially in accordance with FIG. 1
and a differential scanning calorimetry trace substantially in
accordance with FIG. 2 and a thermogravimetric analysis trace
substantially in accordance with FIG. 2. In another embodiment, the
crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern substantially in accordance with FIG. 1
and a differential scanning calorimetry trace substantially in
accordance with FIG. 2 In another embodiment, the crystalline form
of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern substantially in accordance with FIG. 1
and a thermogravimetric analysis trace substantially in accordance
with FIG. 2. In another embodiment, the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern comprising diffraction angles, when
measured using Cu K.sub..alpha. radiation, of about 5.9, 13.6,
14.0, 14.3, 23.3, 24.5, and 27.4 degrees 2.theta., and a
differential scanning calorimetry trace substantially in accordance
with FIG. 2. In another embodiment, the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is characterized by an X-ray powder
diffraction (XRPD) pattern comprising diffraction angles, when
measured using Cu K.sub..alpha. radiation, of about 5.9, 13.6,
14.0, 14.3, 23.3, 24.5, and 27.4 degrees 2.theta., and a
thermogravimetric analysis trace substantially in accordance with
FIG. 2.
[0119] An XRPD pattern will be understood to comprise a diffraction
angle (expressed in degrees 2.theta.) of "about" a value specified
herein when the XRPD pattern comprises a diffraction angle within
.+-.0.3 degrees 2.theta. of the specified value. Further, it is
well known and understood to those skilled in the art that the
apparatus employed, humidity, temperature, orientation of the
powder crystals, and other parameters involved in obtaining an
X-ray powder diffraction (XRPD) pattern may cause some variability
in the appearance, intensities, and positions of the lines in the
diffraction pattern. An X-ray powder diffraction pattern that is
"substantially in accordance" with that of FIG. 1 provided herein
is an XRPD pattern that would be considered by one skilled in the
art to represent a compound possessing the same crystal form as the
compound that provided the XRPD pattern of FIG. 1. That is, the
XRPD pattern may be identical to that of FIG. 1, or more likely it
may be somewhat different. Such an XRPD pattern may not necessarily
show each of the lines of any one of the diffraction patterns
presented herein, and/or may show a slight change in appearance,
intensity, or a shift in position of said lines resulting from
differences in the conditions involved in obtaining the data. A
person skilled in the art is capable of determining if a sample of
a crystalline compound has the same form as, or a different form
from, a form disclosed herein by comparison of their XRPD patterns.
For example, one skilled in the art can overlay an XRPD pattern of
a sample of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid, with FIG. 1 and, using expertise and
knowledge in the art, readily determine whether the XRPD pattern of
the sample is substantially in accordance with the XRPD pattern of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid disclosed herein. If the XRPD pattern is
substantially in accordance with FIG. 1, the sample form can be
readily and accurately identified as having the same form as the
crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo-
[b]thiophen-3-yl)acetic acid disclosed herein.
[0120] The compound of Formula (I) or a salt thereof may exist in
stereoisomeric forms (e.g., it contains 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. It is to be understood that the present
invention includes all combinations and subsets of the particular
groups defined hereinabove. The scope of the present invention
includes mixtures of stereoisomers as well as purified enantiomers
or enantiomerically/diastereomerically enriched mixtures. It is to
be understood that the present invention includes all combinations
and subsets of the particular groups defined hereinabove.
[0121] The subject invention also includes isotopically-labeled
compounds, which are identical to those recited in Formula (I) and
following, but for the fact that one or more atoms are replaced by
an atom having an atomic mass or mass number different from the
atomic mass or mass number usually found in nature. Examples of
isotopes that can be incorporated into compounds of the invention
and pharmaceutically acceptable salts thereof include isotopes of
hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine,
chlorine, and iodine, such as .sup.2H, .sup.3H, .sup.11C, .sup.13C,
.sup.14C, .sup.15N, .sup.17O, .sup.18O, .sup.31P, .sup.32P,
.sup.35S, .sup.18F, .sup.36Cl, .sup.123I, and .sup.125I.
[0122] Compounds of the present invention and pharmaceutically
acceptable salts of said compounds that contain the aforementioned
isotopes and/or other isotopes of other atoms are within the scope
of the present invention. Isotopically-labeled compounds of the
present invention, for example those into which radioactive
isotopes such as .sup.3H, .sup.14C are incorporated, are useful in
drug and/or substrate tissue distribution assays. Tritiated, i.e.,
.sup.3H, and carbon-14, i.e., .sup.14C, isotopes are particularly
preferred for their ease of preparation and detectability. .sup.11C
and .sup.18F isotopes are particularly useful in PET (positron
emission tomography), and .sup.125I isotopes are particularly
useful in SPECT (single photon emission computerized tomography),
all useful in brain imaging. Further, substitution with heavier
isotopes such as deuterium, i.e., .sup.2H, can afford certain
therapeutic advantages resulting from greater metabolic stability,
for example increased in vivo half-life or reduced dosage
requirements and, hence, may be preferred in some circumstances.
Isotopically labeled compounds of Formula (I) and following of this
invention can generally be prepared by carrying out the procedures
disclosed in the Schemes and/or in the Examples below, by
substituting a readily available isotopically labeled reagent for a
non-isotopically labeled reagent.
[0123] The invention further provides a pharmaceutical composition
(also referred to as a pharmaceutical formulation) comprising a
compound of Formula (I) or pharmaceutically acceptable salt thereof
and one or more excipients (also referred to as carriers and/or
diluents in the pharmaceutical arts). The excipients are acceptable
in the sense of being compatible with the other ingredients of the
formulation and not deleterious to the recipient thereof (i.e., the
patient).
[0124] Suitable pharmaceutically acceptable excipients will vary
depending upon the particular dosage form chosen. In addition,
suitable pharmaceutically acceptable excipients may be chosen for a
particular function that they may serve in the composition. For
example, certain pharmaceutically acceptable excipients may be
chosen for their ability to facilitate the production of uniform
dosage forms. Certain pharmaceutically acceptable excipients may be
chosen for their ability to facilitate the production of stable
dosage forms. Certain pharmaceutically acceptable excipients may be
chosen for their ability to facilitate the carrying or transporting
of the compound or compounds of the invention once administered to
the patient from one organ, or portion of the body, to another
organ, or portion of the body. Certain pharmaceutically acceptable
excipients may be chosen for their ability to enhance patient
compliance.
[0125] Suitable pharmaceutically acceptable excipients include the
following types of excipients: diluents, fillers, binders,
disintegrants, lubricants, glidants, granulating agents, coating
agents, wetting agents, solvents, co-solvents, suspending agents,
emulsifiers, sweeteners, flavoring agents, flavor masking agents,
coloring agents, anticaking agents, hemectants, chelating agents,
plasticizers, viscosity increasing agents, antioxidants,
preservatives, stabilizers, surfactants, and buffering agents. The
skilled artisan will appreciate that certain pharmaceutically
acceptable excipients may serve more than one function and may
serve alternative functions depending on how much of the excipient
is present in the formulation and what other ingredients are
present in the formulation.
[0126] Skilled artisans possess the knowledge and skill in the art
to enable them to select suitable pharmaceutically acceptable
excipients in appropriate amounts for use in the invention. In
addition, there are a number of resources that are available to the
skilled artisan which describe pharmaceutically acceptable
excipients and may be useful in selecting suitable pharmaceutically
acceptable excipients. Examples include Remington's Pharmaceutical
Sciences (Mack Publishing Company), The Handbook of Pharmaceutical
Additives (Gower Publishing Limited), and The Handbook of
Pharmaceutical Excipients (the American Pharmaceutical Association
and the Pharmaceutical Press).
[0127] The pharmaceutical compositions of the invention are
prepared using techniques and methods known to those skilled in the
art. Some of the methods commonly used in the art are described in
Remington's Pharmaceutical Sciences (Mack Publishing Company).
[0128] Pharmaceutical compositions may be 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 salt thereof 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 compositions may be
prepared by any of the methods well-known in the pharmacy art.
[0129] In the present invention, tablets and capsules are preferred
for delivery of the pharmaceutical composition.
[0130] In accordance with another aspect of the invention there is
provided a process for the preparation of a pharmaceutical
composition comprising mixing (or admixing) a compound of Formula
(I) or salt thereof with at least one excipient.
[0131] The present invention also provides a method of treatment in
a mammal, especially a human. The compounds and compositions of the
invention are used to treat GOAT mediated disorders or diseases.
Disease states or disorders which can be treated by the methods and
compositions provided herein include, but are not limited to,
obesity
[0132] The present invention also provides a method of treatment in
a subject (e.g., a mammal, especially a human). Disease states or
disorders which can be treated by the methods and compositions or
preparations provided herein include, but are not limited to,
metabolic disorders (e.g. Prader-Willi syndrome, metabolic
syndrome, insulin resistance, impaired glucose tolerance,
prediabetes, diabetes mellitus (e.g., type II diabetes mellitus),
dysglycemia (e.g., hyperglycemia), obesity (e.g., obesity caused by
Prader-Willi syndrome), increased adiposity, poor glycemic control,
hyperphagia, impaired satiety, dyslipidemia (e.g., atherogenic
dyslipidemia), hepatic steatosis (e.g., non-alcoholic fatty liver
disease (e.g., non-alcoholic steatohepatitis))), psychiatric
disorders (e.g., eating disorders (e.g., bulimia nervosa, binge
eating disorder, night-time eating syndrome), substance related
disorders (e.g., addiction disorders (e.g., alcohol, smoking,
overeating, or use of illicit drugs))), as well as disorders
related to or complications of metabolic or psychiatric disorders
(e.g., cardiovascular diseases (e.g., diabetic heart disease (e.g.,
diabetic cardiomyopathy), heart failure, or hypertension), ischemia
(e.g., myocardial ischemia, cerebral ischemia, ischemic stroke), or
BMI-related cancers (e.g., pancreatic cancer, gallbladder cancer,
esophageal cancer, colorectal cancer, breast cancer etc.).
[0133] The compositions and methods provided herein are
particularly deemed useful for the treatment of GOAT mediated
disorders, such as obesity, increased adiposity, poor glycemic
control, etc. The compositions and methods provided herein are
particularly deemed useful for the treatment of GOAT mediated
disorders, such as metabolic disorders (e.g. Prader-Willi syndrome,
metabolic syndrome, insulin resistance, impaired glucose tolerance,
prediabetes, diabetes mellitus (e.g., type II diabetes mellitus),
dysglycemia (e.g., hyperglycemia), obesity (e.g., obesity caused by
Prader-Willi syndrome), increased adiposity, poor glycemic control,
hyperphagia, impaired satiety, dyslipidemia (e.g., atherogenic
dyslipidemia), hepatic steatosis (e.g., non-alcoholic fatty liver
disease (e.g., non-alcoholic steatohepatitis))), psychiatric
disorders (e.g., eating disorders (e.g., bulimia nervosa, binge
eating disorder, night-time eating syndrome), substance related
disorders (e.g., addiction disorders (e.g., alcohol, smoking,
overeating, or use of illicit drugs))), as well as disorders
related to or complications of metabolic or psychiatric disorders
(e.g., cardiovascular diseases (e.g., diabetic heart disease (e.g.,
diabetic cardiomyopathy), heart failure, or hypertension), ischemia
(e.g., myocardial ischemia, cerebral ischemia, ischemic stroke), or
BMI-related cancers (e.g., pancreatic cancer, gallbladder cancer,
esophageal cancer, colorectal cancer, breast cancer etc.)). More
particularly, diseases or disorders that may be treated by the
compositions and methods of the invention include Prader-Willi
syndrome, excess weight, and/or obesity (e.g., obesity caused by
Prader-Willi syndrome). Weight that is higher than what is
considered as a healthy weight for a given height is considered
overweight or obese. In one embodiment, a compound of the invention
is administered to a human having a body mass index (BMI) of at
least about 25. In one embodiment, a compound of the invention is
administered to a human having a body mass index (BMI) of at least
about 26. In one embodiment, a compound of the invention is
administered to a human having a body mass index (BMI) of at least
about 27. In one embodiment, a compound of the invention is
administered to a human having a body mass index (BMI) of at least
about 28. In one embodiment, a compound of the invention is
administered to a human having a body mass index (BMI) of at least
about 29. In one embodiment, a compound of the invention is
administered to a human having a body mass index (BMI) of at least
about 30. In another embodiment, a compound of the invention is
administered to a human having a body mass index (BMI) of at least
about 31, at least about 32, at least about 33, at least about 34,
at least about 35, at least about 36, at least about 37, at least
about 38, at least about 39, or at least about 40. In one
embodiment, the obesity is extreme or severe obesity. In a
particular embodiment, the obesity is caused by Prader-Willi
syndrome.
[0134] The instant compounds can be combined with or
co-administered with other therapeutic agents, particularly agents
that may enhance the activity or time of disposition of the
compounds. Combination therapies according to the invention
comprise the administration of at least one compound of the
invention and the use of at least one other treatment method. In
one embodiment, combination therapies according to the invention
comprise the administration of at least one compound of the
invention and surgical therapy, such as bariatric surgery. In one
embodiment, combination therapies according to the invention
comprise the administration of at least one compound of the
invention and lifestyle modification. Lifestyle modification can
include, for example, a reduced-calorie diet and/or exercise. In
one embodiment, combination therapies according to the invention
comprise the administration of at least one compound of the
invention and a weight-loss agent, such as orlistat, lorcaserin,
liraglutide, phentermine/topimarate, or naltrexone/bupropion. In
one embodiment, combination therapies according to the invention
comprise the administration of at least one compound of the
invention and a hormone therapy (e.g., testosterone, estrogen,
progesterone, or human growth hormone), selective serotonin
reuptake inhibitors (SSRIs), or anti-diabetic therapy (e.g.,
insulin, miglitol, acarbose, metformin, exenatide, pramlintide). In
yet another embodiment, the invention comprises a therapeutic
regimen where the GOAT inhibitors of this disclosure are not in and
of themselves active or significantly active, but when combined
with another therapy, which may or may not be active as a
standalone therapy, the combination provides a useful therapeutic
outcome.
[0135] By the term "co-administration" and derivatives thereof as
used herein refers to either simultaneous administration or any
manner of separate sequential administration of a GOAT inhibiting
compound, as described herein, and a further active ingredient or
ingredients, known to be useful in the treatment of obesity,
including orlistat, lorcaserin, liraglutide,
phentermine/topimarate, and naltrexone/bupropion, or a hormone
therapy (e.g., testosterone, estrogen, progesterone, or human
growth hormone), selective serotonin reuptake inhibitors (SSRIs),
or anti-diabetic therapy (e.g., insulin, miglitol, acarbose,
metformin, exenatide, pramlintide). The term further active
ingredient or ingredients, as used herein, includes any compound or
therapeutic agent known to or that demonstrates advantageous
properties when administered to a patient in need of treatment for
obesity. Preferably, if the administration is not simultaneous, the
compounds are administered in a close time proximity to each other.
Furthermore, it does not matter if the compounds are administered
in the same dosage form, e.g. one compound may be administered
topically and another compound may be administered orally.
[0136] Typically, any weight loss agent may be co-administered in
the treatment of obesity in the present invention. Typically, any
weight loss agent, hormone therapy, or anti-diabetic therapy may be
co-administered in the methods and uses of the present invention. A
person of ordinary skill in the art would be able to discern which
combinations of agents would be useful based on the particular
characteristics of the drugs and the obesity involved. Typical
weight loss agents useful in the present invention include, but are
not limited to, appetite-suppressing agents and lipase
inhibitors.
[0137] Examples of a further active ingredient or ingredients for
use in combination or co-administered with the present GOAT
inhibiting compounds are weight-loss agents. Examples of
weight-loss agents include, but are not limited to, orlistat,
lorcaserin, liraglutide, phentermine/topimarate, and
naltrexone/bupropion.
[0138] Orlistat is a lipase inhibitor which prevents some of the
fat in foods eaten from being absorbed in the intestines. The
unabsorbed fat is removed from the body in the stool.
[0139] Lorcaserin (BELVIQ) is a serotonin receptor agonist.
Lorcaserin targets the 5HT2C receptor and alters body weight by
regulating satiety.
[0140] Liraglutide (SAXENDA) is a glucagonlike peptide-1 (GLP-1)
receptor agonist. Liraglutide is an anti-diabetic agent that has
been approved for weight loss.
[0141] Phentermine/Topimarate (QYSMIA) is a combination product.
Phentermine is an anorectic and topiramate is an anticonvulsant.
Phentermine/Topimarate decreases appetite and causes feelings of
fullness to last longer after eating.
[0142] Naltrexone/Bupropion (CONTRAVE) is a combination product.
Naltrexone is an opiate antagonist and Bupropion is an
antidepressant. Naltrexone/Bupropion regulates brain activity to
reduce appetite.
[0143] Pharmaceutical compositions may be presented in unit dose
forms containing a predetermined amount of active ingredient per
unit dose. Such a unit may contain, for example, 0.5 mg to 1 g,
preferably 1 mg to 700 mg, more preferably 5 mg to 100 mg of a
compound of the Formula (I), depending on the condition being
treated, the route of administration and the age, weight and
condition of the patient, or pharmaceutical compositions may be
presented in unit dose forms containing a predetermined amount of
active ingredient per unit dose. Preferred unit dosage compositions
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 compositions may be
prepared by any of the methods well known in the pharmacy art.
[0144] Pharmaceutical compositions 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 compositions may be prepared by any method known in the
art of pharmacy, for example by bringing into association a
compound of formula (I) with the carrier(s) or excipient(s).
[0145] Pharmaceutical compositions 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.
[0146] 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.
[0147] 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, carboxymethylcellulose, 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 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.
[0148] 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 a compound of Formula (I). 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.
[0149] Where appropriate, dosage unit pharmaceutical compositions
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.
[0150] Pharmaceutical compositions adapted for rectal
administration may be presented as suppositories or as enemas.
[0151] Pharmaceutical compositions adapted for vaginal
administration may be presented as pessaries, tampons, creams,
gels, pastes, foams or spray formulations.
[0152] Pharmaceutical formulations adapted for parenteral
administration include aqueous and non-aqueous sterile injection
solutions which may contain anti-oxidants, buffers, bacteriostats
and solutes which render the composition isotonic with the blood of
the intended recipient; and aqueous and non-aqueous sterile
suspensions which may include suspending agents and thickening
agents. The pharmaceutical 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.
[0153] It should be understood that in addition to the ingredients
particularly mentioned above, the pharmaceutical 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 flavoring agents.
[0154] A therapeutically effective amount of a compound of the
present invention will depend upon a number of factors including,
for example, the age and weight of the intended recipient, the
precise condition requiring treatment and its severity, the nature
of the formulation, and the route of administration, and will
ultimately be at the discretion of the attendant prescribing the
medication. However, an effective amount of a compound of Formula
(I) for the treatment of obesity will generally be in the range of
0.001 to 100 mg/kg body weight of recipient per day, suitably in
the range of 0.01 to 10 mg/kg body weight per day. For a 70 kg
adult mammal, the actual amount per day would suitably be from 7 to
700 mg and this amount may be given in a single dose per day or in
a number (such as two, three, four, five or six) of sub-doses per
day such that the total daily dose is the same. An effective amount
of a salt or solvate, etc., may be determined as a proportion of
the effective amount of the compound of Formula (I) per se. It is
envisaged that similar dosages would be appropriate for treatment
of the other conditions referred to above.
[0155] In certain embodiments, this invention relates to a
pharmaceutical composition comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid. In another embodiment, this invention
relates to a pharmaceutical composition comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid wherein at least 10% by weight of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is present as the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid described herein. In another embodiment,
this invention relates to a pharmaceutical composition comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid wherein at least 20% by weight, or at
least 30% by weight, or at least 40% by weight, or at least 50% by
weight, or at least 60% by weight, or at least 70% by weight, or at
least 80% by weight, or at least 90% by weight of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is present as the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid described herein. In another embodiment,
this invention relates to a pharmaceutical composition comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid wherein at least 95% by weight, or at
least 96% by weight, or at least 97% by weight, or at least 98% by
weight, or at least 99% by weight, or at least 99.5% by weight, or
at least 99.8% by weight, or at least 99.9% by weight of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is present as the crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid described herein.
[0156] In another embodiment, this invention relates to a
pharmaceutical composition comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid, wherein not more than 90% by weight of
the compound is amorphous. In another embodiment, this invention
relates to a pharmaceutical composition comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid, wherein not more than 80% by weight, or
not more than 70% by weight, or not more than 60% by weight, or not
more than 50% by weight, or not more than 40% by weight, or not
more than 30% by weight, or not more than 20% by weight, or not
more than 10% by weight of the compound is amorphous. In another
embodiment, this invention relates to a pharmaceutical composition
comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid, wherein not more than 5% by weight, or
not more than 4% by weight, or not more than 3% by weight, or not
more than 2% by weight, or not more than 1% by weight, or not more
than 0.5% by weight, or not more than 0.2% by weight, or not more
than 0.1% by weight of the compound is amorphous.
[0157] In another embodiment, this invention relates to a
pharmaceutical composition comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid wherein not more than 90% by weight of
the
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is present in a form other than the
crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-
benzo[b]thiophen-3-yl)acetic acid described herein. In another
embodiment, this invention relates to a pharmaceutical composition
comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid wherein not more than 80% by weight, or
not more than 70% by weight, or not more than 60% by weight, or not
more than 50% by weight, or not more than 40% by weight, or not
more than 30% by weight, or not more than 20% by weight, or not
more than 10% by weight of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is present in a form other than the
crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-
benzo[b]thiophen-3-yl)acetic acid described herein. In another
embodiment, this invention relates to a pharmaceutical composition
comprising
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid wherein not more than 5% by weight, or
not more than 4% by weight, or not more than 3% by weight, or not
more than 2% by weight, or not more than 1% by weight, or not more
than 0.5% by weight, or not more than 0.2% by weight, or not more
than 0.1% by weight of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid is present in a form other than the
crystalline form of
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-
benzo[b]thiophen-3-yl)acetic acid described herein.
Definitions
[0158] Terms are used within their accepted meanings. The following
definitions are meant to clarify, but not limit, the terms
defined.
[0159] As used herein, the term "alkyl" represents a saturated,
straight or branched hydrocarbon moiety having the specified number
of carbon atoms. The term "(C.sub.1-C.sub.4)alkyl" refers to an
alkyl moiety containing from 1 to 4 carbon atoms. Exemplary alkyls
include, but are not limited to, methyl, ethyl, n-propyl,
isopropyl, n-butyl, isobutyl, s-butyl, and t-butyl.
[0160] The term "halo(C.sub.1-C.sub.4)alkyl" is intended to mean a
radical having one or more halogen atoms, which may be the same or
different, at one or more carbon atoms of an alkyl moiety
containing from 1 to 4 carbon atoms, which is a straight or
branched-chain carbon radical. Examples of
"halo(C.sub.1-C.sub.4)alkyl" groups useful in the present invention
include, but are not limited to, --CF.sub.3 (trifluoromethyl),
--CCl.sub.3 (trichloromethyl), 1,1-difluoroethyl,
2-fluoro-2-methylpropyl, 2,2-difluoropropyl, 2,2,2-trifluoroethyl,
and hexafluoroisopropyl.
[0161] The terms "halogen" and "halo" represent fluoro, chloro,
bromo, or iodo substituents.
[0162] As used herein, the term "cyano" refers to the group
--CN.
[0163] In certain embodiments, the substituent present on the
nitrogen atom is an nitrogen protecting group (also referred to
herein as an "amino protecting group"). Nitrogen protecting groups
include, but are not limited to, --OH, --ORaa, --N(Rcc)2,
--C(.dbd.O)Raa, --C(.dbd.O)N(Rcc)2, --CO2Raa, --SO2Raa,
--C(.dbd.NRcc)Raa, --C(.dbd.NRcc)ORaa, --C(.dbd.NRcc)N(Rcc)2,
--SO2N(Rcc)2, --SO2Rcc, --SO2ORcc, --SORaa, --C(.dbd.S)N(Rcc)2,
--C(.dbd.O)SRcc, --C(.dbd.S)SRcc, C1-10 alkyl (e.g., aralkyl,
heteroaralkyl), C2-10 alkenyl, C2-10 alkynyl, heteroC1-10alkyl,
heteroC2-10alkenyl, heteroC2-10alkynyl, C3-10 carbocyclyl, 3-14
membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl
groups, wherein each alkyl, alkenyl, alkynyl, heteroalkyl,
heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aralkyl,
aryl, and heteroaryl is independently substituted with 0, 1, 2, 3,
4, or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as
described herein. Nitrogen protecting groups are well known in the
art and include those described in detail in Protecting Groups in
Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition,
John Wiley & Sons, 1999, incorporated herein by reference.
[0164] For example, nitrogen protecting groups such as amide groups
(e.g., --C(.dbd.O)Raa) include, but are not limited to, formamide,
acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide,
phenylacetamide, 3-phenylpropanamide, picolinamide,
3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide,
p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide,
acetoacetamide, (N'-dithiobenzyloxyacylamino)acetamide,
3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide,
2-methyl-2-(o-nitrophenoxy)propanamide,
2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,
3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine
derivative, o-nitrobenzamide and o-(benzoyloxymethyl)benzamide.
[0165] Nitrogen protecting groups such as carbamate groups (e.g.,
--C(.dbd.O)ORaa) include, but are not limited to, methyl carbamate,
ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc),
9-(2-sulfo)fluorenylmethyl carbamate,
9-(2,7-dibromo)fluoroenylmethyl carbamate,
2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl
carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),
2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl
carbamate (Teoc), 2-phenylethyl carbamate (hZ),
1-(1-adamantyl)-1-methylethyl carbamate (Adpoc),
1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl
carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate
(TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),
1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'-
and 4'-pyridyl)ethyl carbamate (Pyoc),
2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate
(BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl
carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl
carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl
carbamate, N-hydroxypiperidinyl carbamate, alkyldithio carbamate,
benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),
p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl
carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl
carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl
carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl
carbamate, 2-(p-toluenesulfonyl)ethyl carbamate,
[2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl
carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc),
2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl
carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate,
m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl
carbamate, 5-benzisoxazolylmethyl carbamate,
2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc),
m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate,
o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate,
phenyl(o-nitrophenyl)methyl carbamate, t-amyl carbamate, S-benzyl
thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate,
cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl
carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl
carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate,
1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,
1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,
2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl
carbamate, isobutyl carbamate, isonicotinyl carbamate,
p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl
carbamate, 1-methylcyclohexyl carbamate,
1-methyl-1-cyclopropylmethyl carbamate,
1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,
1-methyl-1-(p-phenylazophenyl)ethyl carbamate,
1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl
carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate,
2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl
carbamate, and 2,4,6-trimethylbenzyl carbamate.
[0166] Nitrogen protecting groups such as sulfonamide groups (e.g.,
--S(.dbd.O)2Raa) include, but are not limited to,
p-toluenesulfonamide (Ts), benzenesulfonamide,
2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),
2,4,6-trimethoxybenzenesulfonamide (Mtb),
2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),
2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),
4-methoxybenzenesulfonamide (Mbs),
2,4,6-trimethylbenzenesulfonamide (Mts),
2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),
2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc),
methanesulfonamide (Ms), .beta.-trimethylsilylethanesulfonamide
(SES), 9-anthracenesulfonamide,
4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),
benzylsulfonamide, trifluoromethylsulfonamide, and
phenacylsulfonamide.
[0167] Other nitrogen protecting groups include, but are not
limited to, phenothiazinyl-(10)-acyl derivative,
N'-p-toluenesulfonylaminoacyl derivative, N'-phenylaminothioacyl
derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine
derivative, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide,
N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide,
N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane
adduct (STABASE), 5-substituted
1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted
1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted
3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,
N-[2-(trimethylsilyl)ethoxy]methylamine (SEM),
N-3-acetoxypropylamine,
N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary
ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,
N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),
N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),
N-9-phenylfluorenylamine (PhF),
N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino
(Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine,
N-benzylideneamine, N-p-methoxybenzylideneamine,
N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,
N--(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine,
N-p-nitrobenzylideneamine, N-salicylideneamine,
N-5-chlorosalicylideneamine,
N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,
N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,
N-borane derivative, N-diphenylborinic acid derivative,
N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper
chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine
N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide
(Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates,
dibenzyl phosphoramidate, diphenyl phosphoramidate,
benzenesulfenamide, o-nitrobenzenesulfenamide (Nps),
2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide,
2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide,
and 3-nitropyridinesulfenamide (Npys). In certain embodiments, a
nitrogen protecting group is benzyl (Bn), tert-butyloxycarbonyl
(BOC), carbobenzyloxy (Cbz), 9-flurenylmethyloxycarbonyl (Fmoc),
trifluoroacetyl, triphenylmethyl, acetyl (Ac), benzoyl (Bz),
p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl
(PMP), 2,2,2-trichloroethyloxycarbonyl (Troc), triphenylmethyl
(Tr), tosyl (Ts), brosyl (Bs), nosyl (Ns), mesyl (Ms), triflyl
(Tf), or dansyl (Ds).
[0168] "Pharmaceutically acceptable" refers to those compounds,
materials, compositions, and dosage forms which are, within the
scope of sound medical judgment, suitable for use in contact with
the tissues of human beings and animals without excessive toxicity,
irritation, or other problem or complication, commensurate with a
reasonable benefit/risk ratio.
[0169] As used herein, the term "pharmaceutically acceptable salts"
refers to salts that retain the desired biological activity of the
subject compound. These pharmaceutically acceptable salts may be
prepared in situ during the final isolation and purification of the
compound, or by separately reacting the purified compound in its
free acid or free base form with a suitable base or acid,
respectively. Examples of pharmaceutically acceptable salts include
sulfates, pyrosulfates, bisulfates, sulfites, bisulfites,
phosphates, chlorides, bromides, iodides, acetates, propionates,
decanoates, caprylates, acrylates, formates, isobutyrates,
caproates, heptanoates, propiolates, oxalates, malonates
succinates, suberates, sebacates, fumarates, maleates,
butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates,
methylbenzoates, dinitrobenzoates, hydroxybenzoates,
methoxybenzoates, phthalates, phenylacetates, phenylpropionates,
phenylbutrates, citrates, lactates, 7-hydroxybutyrates, glycolates,
tartrates mandelates, and sulfonates, such as xylenesulfonates,
methanesulfonates, propanesulfonates, naphthalene-1-sulfonates and
naphthalene-2-sulfonates. Salts of the disclosed compounds
containing a carboxylic acid or other acidic functional group can
be prepared by reacting with a suitable base. Such a
pharmaceutically acceptable salt may be made with a base which
affords a pharmaceutically acceptable cation, which includes alkali
metal salts (especially sodium and potassium), alkaline earth metal
salts (especially calcium and magnesium), aluminum salts and
ammonium salts, as well as salts made from physiologically
acceptable organic bases such as trimethylamine, triethylamine,
morpholine, pyridine, piperidine, picoline, dicyclohexylamine,
N,N'-dibenzylethylenediamine, 2-hydroxyethylamine,
bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine,
dibenzylpiperidine, dehydroabietylamine,
N,N'-bisdehydroabietylamine, glucamine, N-methylglucamine,
collidine, quinine, quinoline, and basic amino acid such as lysine
and arginine.
[0170] As used herein, the term "compound(s) of the invention"
means a compound of Formula (I) (as defined above) in any form,
i.e., any salt or non-salt form (e.g., as a free acid or base form,
or as a pharmaceutically acceptable salt thereof) and any physical
form thereof (e.g., including non-solid forms (e.g., liquid or
semi-solid forms), and solid forms (e.g., amorphous or crystalline
forms, specific polymorphic forms, solvates, including hydrates
(e.g., mono-, di- and hemi-hydrates)), and mixtures of various
forms.
[0171] As used herein, the terms "treatment", "treat," and
"treating" refer to reversing, alleviating the specified condition,
eliminating or reducing one or more symptoms of the condition,
delaying the onset of, slowing or eliminating the progression of
the condition, and delaying the reoccurrence of a condition in a
previously afflicted or diagnosed patient or subject. In some
embodiments, treatment may be administered after one or more signs
or symptoms of the disease have developed or have been observed. In
other embodiments, treatment may be administered in the absence of
signs or symptoms of the disease. For example, treatment may be
administered to a susceptible subject prior to the onset of
symptoms (e.g., in light of a history of symptoms and/or in light
of exposure to a pathogen). Treatment may also be continued after
symptoms have resolved, for example, to delay or prevent
recurrence.
[0172] A "subject" to which administration is contemplated refers
to a human (i.e., male or female of any age group, e.g., paediatric
subject (e.g., infant, child, or adolescent) or adult subject
(e.g., young adult, middle-aged adult, or senior adult)) or
non-human animal. In certain embodiments, the non-human animal is a
mammal (e.g., primate (e.g., cynomolgus monkey or rhesus monkey),
commercially relevant mammal (e.g., cattle, pig, horse, sheep,
goat, cat, or dog). The non-human animal may be a transgenic animal
or genetically engineered animal. The term "patient" refers to a
human subject in need of treatment of a disease.
[0173] The term "biological sample" refers to any sample including
tissue samples (such as tissue sections and needle biopsies of a
tissue); cell samples (e.g., cytological smears (such as Pap or
blood smears) or samples of cells obtained by microdissection);
samples of whole organisms (such as samples of yeasts or bacteria);
or cell fractions, fragments or organelles (such as obtained by
lysing cells and separating the components thereof by
centrifugation or otherwise). Other examples of biological samples
include blood, serum, urine, semen, fecal matter, cerebrospinal
fluid, interstitial fluid, mucous, tears, sweat, pus, biopsied
tissue (e.g., obtained by a surgical biopsy or needle biopsy),
nipple aspirates, milk, vaginal fluid, saliva, swabs (such as
buccal swabs), or any material containing biomolecules that is
derived from a first biological sample.
[0174] The terms "condition," "disease," and "disorder" are used
interchangeably.
[0175] As used herein, the term "effective amount" means that
amount of a drug or pharmaceutical agent that will elicit the
biological or medical response of a tissue, system, animal, or
human that is being sought, for instance, by a researcher or
clinician. An effective amount of a compound described herein may
vary depending on such factors as the desired biological endpoint,
the pharmacokinetics of the compound, the condition being treated,
the mode of administration, and the age and health of the subject.
In certain embodiments, an effective amount is a therapeutically
effective amount. In certain embodiments, an effective amount is a
prophylactic treatment. In certain embodiments, an effective amount
is the amount of a compound described herein in a single dose. In
certain embodiments, an effective amount is the combined amounts of
a compound described herein in multiple doses.
[0176] The term "therapeutically effective amount" of a compound
described herein is any amount sufficient to provide a therapeutic
benefit in the treatment of a condition or to delay or minimize one
or more symptoms associated with the condition. A therapeutically
effective amount of a compound means an amount of therapeutic
agent, alone or in combination with other therapies, which provides
a therapeutic benefit in the treatment of the condition as compared
to a corresponding subject who has not received such amount,
resulting in improved treatment, healing, or amelioration of a
disease, disorder, or side effect, or a decrease in the rate of
advancement of a disease or disorder. The term also includes within
its scope amounts effective to enhance normal physiological
function. For use in therapy, therapeutically effective amounts of
a compound of Formula (I), as well as salts thereof, may be
administered as the raw chemical. Additionally, the active
ingredient may be presented as a pharmaceutical composition or
preparation. Additionally, the active ingredient or salt thereof
may be presented as a pharmaceutical composition or preparation.
The term "therapeutically effective amount" can encompass an amount
that improves overall therapy, reduces or avoids symptoms, signs,
or causes of the condition, and/or enhances the therapeutic
efficacy of another therapeutic agent. In certain embodiments, a
therapeutically effective amount is an amount sufficient for
inhibition of GOAT in a subject, biological sample, tissue, or
cell.
[0177] As used herein the term "inhibit" or "inhibition" in the
context of proteins, for example, in the context of GOAT, refers to
a reduction in the activity of the enzyme. In some embodiments, the
term refers to a reduction of the level of activity, e.g., GOAT
activity, to a level that is statistically significantly lower than
an initial level, which may, for example, be a baseline level of
activity. In some embodiments, the term refers to a reduction of
the level of enzyme activity, e.g., GOAT activity, to a level that
is less than 75%, less than 50%, less than 40%, less than 30%, less
than 25%, less than 20%, less than 10%, less than 9%, less than 8%,
less than 7%, less than 6%, less than 5%, less than 4%, less than
3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%,
less than 0.01%, less than 0.001%, or less than 0.0001% of an
initial level, which may, for example, be a baseline level of
enzyme activity.
[0178] Pharmaceutical compositions or preparations described herein
can be prepared by any method known in the art of pharmacology. In
general, such preparatory methods include bringing the compound
described herein (i.e., the "active ingredient") into association
with a carrier or excipient, and/or one or more other accessory
ingredients, and then, if necessary and/or desirable, shaping,
and/or packaging the product into a desired single- or multi-dose
unit.
[0179] The term "metabolic disorder" refers to any disorder that
involves an alteration in the normal metabolism of carbohydrates,
lipids, proteins, nucleic acids, or a combination thereof. A
metabolic disorder is associated with either a deficiency or excess
in a metabolic pathway resulting in an imbalance in metabolism of
nucleic acids, proteins, lipids, and/or carbohydrates. Factors
affecting metabolism include, and are not limited to, the endocrine
(hormonal) control system (e.g., the insulin pathway, the
enteroendocrine hormones including GLP-1, PYY or the like), the
neural control system (e.g., GLP-1 in the brain), or the like.
Examples of metabolic disorders include, but are not limited to,
Prader-Willi syndrome, metabolic syndrome, insulin resistance,
impaired glucose tolerance, prediabetes, diabetes mellitus (e.g.,
type II diabetes mellitus), dysglycemia (e.g., hyperglycemia),
obesity (e.g., obesity caused by Prader-Willi syndrome), increased
adiposity, poor glycemic control, hyperphagia, impaired satiety,
dyslipidemia (e.g., atherogenic dyslipidemia), hepatic steatosis
(e.g., non-alcoholic fatty liver disease (e.g., non-alcoholic
steatohepatitis))).
[0180] A "diabetic condition" refers to diabetes and pre-diabetes.
Diabetes refers to a group of metabolic diseases in which a person
has high blood sugar, either because the body does not produce
enough insulin, or because cells do not respond to the insulin that
is produced. This high blood sugar produces the classical symptoms
of polyuria (frequent urination), polydipsia (increased thirst) and
polyphagia (increased hunger). There are several types of diabetes.
Type I diabetes results from the body's failure to produce insulin,
and presently requires the person to inject insulin or wear an
insulin pump. Type II diabetes results from insulin resistance a
condition in which cells fail to use insulin properly, sometimes
combined with an absolute insulin deficiency. Gestational diabetes
occurs when pregnant women without a previous diagnosis of diabetes
develop a high blood glucose level. Other forms of diabetes include
congenital diabetes, which is due to genetic defects of insulin
secretion, cystic fibrosis-related diabetes, steroid diabetes
induced by high doses of glucocorticoids, and several forms of
monogenic diabetes, e.g., mature onset diabetes of the young (e.g.,
MODY 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). Pre-diabetes indicates a
condition that occurs when a person's blood glucose levels are
higher than normal but not high enough for a diagnosis of diabetes.
All forms of diabetes increase the risk of long-term complications.
These typically develop after many years, but may be the first
symptom in those who have otherwise not received a diagnosis before
that time. The major long-term complications relate to damage to
blood vessels. Diabetes doubles the risk of cardiovascular disease
and macrovascular diseases such as ischemic heart disease (angina,
myocardial infarction), stroke, and peripheral vascular disease.
Diabetes also causes microvascular complications, e.g., damage to
the small blood vessels. Diabetic retinopathy, which affects blood
vessel formation in the retina of the eye, can lead to visual
symptoms, reduced vision, and potentially blindness. Diabetic
nephropathy, the impact of diabetes on the kidneys, can lead to
scarring changes in the kidney tissue, loss of small or
progressively larger amounts of protein in the urine, and
eventually chronic kidney disease requiring dialysis. Diabetic
neuropathy is the impact of diabetes on the nervous system, most
commonly causing numbness, tingling and pain in the feet and also
increasing the risk of skin damage due to altered sensation.
Together with vascular disease in the legs, neuropathy contributes
to the risk of diabetes-related foot problems, e.g., diabetic foot
ulcers, that can be difficult to treat and occasionally require
amputation.
[0181] The term "psychiatric disorder" refers to a disease of the
mind and includes diseases and disorders listed in the Diagnostic
and Statistical Manual of Mental Disorders--Fourth Edition
(DSM-IV), published by the American Psychiatric Association,
Washington D. C. (1994). Psychiatric disorders include, but are not
limited to, eating disorders (e.g., night eating syndrome),
substance-related disorders (e.g., alcohol dependence, amphetamine
dependence, cannabis dependence, cocaine dependence, hallucinogen
dependence, inhalant dependence, nicotine dependence, opioid
dependence, phencyclidine dependence, and sedative dependence).
[0182] An "obesity-related condition" includes, but is not limited
to, Prader-Willi syndrome, obesity, undesired weight gain (e.g.,
from medication-induced weight gain, from cessation of smoking) and
an over-eating disorder (e.g., binge eating, bulimia, compulsive
eating, or a lack of appetite control each of which can optionally
lead to undesired weight gain or obesity). "Obesity" and "obese"
refers to class I obesity, class II obesity, class III obesity, and
pre-obesity (e.g., being "over-weight") as defined by the World
Health Organization.
[0183] Reduction of storage fat is expected to provide various
primary and/or secondary benefits in a subject (e.g., in a subject
diagnosed with a complication associated with obesity) such as, for
example, an increased insulin responsiveness (e.g., in a subject
diagnosed with Type II diabetes mellitus); a reduction in elevated
blood pressure; a reduction in elevated cholesterol levels; and/or
a reduction (or a reduced risk or progression) of ischemia (e.g.,
ischemic heart disease, cerebral ischemia, or ischemic stroke)
arterial vascular disease, angina, myocardial infarction, stroke,
migraines, congestive heart failure, deep vein thrombosis,
pulmonary embolism, gall stones, gastroesophagael reflux disease,
obstructive sleep apnea, obesity hypoventilation syndrome, asthma,
gout, poor mobility, back pain, erectile dysfunction, urinary
incontinence, liver injury (e.g., fatty liver disease, liver
cirrhosis, alcoholic cirrhosis, endotoxin mediated liver injury) or
chronic renal failure. Thus, the method of this invention is
applicable to obese subjects, diabetic subjects, and alcoholic
subjects.
Compound Preparation
Abbreviations
[0184] AcOEt ethyl acetate [0185] AcOH acetic acid [0186] ADDP
1,1'-(azodicarbonyl)dipiperidine [0187] Ar gas [0188] aq aqueous
[0189] BBr.sub.3 boron tribromide [0190] Boc tert-butyloxycarbonyl
[0191] Bu.sub.4NCl tetrabutylammonium chloride [0192] CDCl.sub.3
deuterochloroform [0193] CHAPS
3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate
[0194] CH.sub.3CN acetonitrile [0195] Cs.sub.2CO.sub.3 cesium
carbonate [0196] DCE 1,2-dichloroethane [0197] DCM dichloromethane
[0198] DIAD diisopropyl azodicarboxylate [0199] DIPEA
N,N-diisopropylethylamine [0200] DM water demineralized water
[0201] DMA dimethylacetamide [0202] DME 1,2-dimethoxyethane [0203]
DMF N,N-dimethylformamide [0204] DMSO dimethyl sulfoxide [0205]
DMPU N,N'-dimethylpropylene urea [0206] EDC
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide [0207] EDTA
ethylenediaminetetraacetic acid [0208] ES electrospray [0209]
Et.sub.3N triethylamine [0210] Et.sub.2O diethyl ether [0211] EtOAc
ethyl acetate [0212] EtOH ethanol [0213] h hour(s) [0214] H.sub.2
hydrogen gas [0215] HCl hydrochloric acid [0216] H.sub.2O water
[0217] HOAt 1-hydroxy-7-azabenzotriazole [0218] H.sub.2SO.sub.4
sulfuric acid [0219] HPLC high-performance liquid chromatography
[0220] HTRF homogeneous time resolved fluorescence [0221] KOtBu
potassium tert-butoxide [0222] K.sub.2CO.sub.3 potassium carbonate
[0223] KMnO.sub.4 potassium permanganate [0224] LCMS liquid
chromatography mass spectrometry [0225] LiAlH.sub.4 lithium
aluminum hydride [0226] LiOH lithium hydroxide [0227] MeI methyl
iodide [0228] MeOH methanol [0229] MeSO.sub.3H methanesulfonic acid
[0230] MgSO.sub.4 magnesium sulfate [0231] MOPS
3-(N-morpholino)propanesulfonic acid [0232] min minute(s) [0233] M
molar [0234] MS mass spectrometry [0235] MTBE methyl tert-butyl
ether [0236] N normal [0237] N.sub.2 nitrogen gas [0238] NaBH.sub.4
sodium borohydride [0239] NaBH(OAc).sub.3 sodium
triacetoxyborohydride [0240] Na.sub.2CO.sub.3 sodium carbonate
[0241] NaH sodium hydride [0242] NaHCO.sub.3 sodium bicarbonate
[0243] NaHMDS sodium bis(trimethylsilyl)amide [0244] NaOH sodium
hydroxide [0245] NaOMe sodium methoxide [0246] Na.sub.2SO.sub.4
sodium sulfate [0247] (n-Bu).sub.3P tri-n-butylphosphine [0248] NBS
N-bromosuccinimide [0249] NH.sub.4Cl ammonium chloride [0250]
NH.sub.4OAc ammonium acetate [0251] NH.sub.4OH ammonium hydroxide
[0252] NMM N-methylmorpholine [0253] Pd--C palladium on carbon
[0254] [PdCl(allyl)].sub.2 allylpalladium(II) chloride dimer [0255]
Pd(OAc).sub.2 palladium(II) acetate [0256] Pd(PPh.sub.3).sub.4
tetrakis(triphenylphosphine)palladium(0) [0257] Pd.sub.2(dba).sub.3
tris(dibenzylideneacetone)dipalladium(0) [0258] PLM polarized light
microscopy [0259] Pet ether petroleum ether [0260] P(o-tol).sub.3
tri(o-tolyl)phosphine [0261] POBr.sub.3 phosphorus(V) oxybromide
[0262] RB round bottom [0263] RT or r.t. room temperature [0264]
RuCl[(R,R)-Tsdpen](mesitylene)
[N-[(1R,2R)-2-(Amino-.kappa.N)-1,2-diphenylethyl]-4-methylbenzenesulfonam-
idato-N]chloro
[(1,2,3,4,5,6-.eta.)-1,3,5-trimethylbenzene]-ruthenium [0265] sat.
saturated [0266] SFC supercritical fluid chromatography [0267]
SOCl.sub.2 thionyl chloride [0268] tBuOH tert-butanol [0269] TBME
tert-butyl methyl ether [0270] TEA triethylamine [0271] TFA
trifluoroacetic acid [0272] THF tetrahydrofuran [0273] TLC thin
layer chromatography [0274] TRF time resolved fluorescence [0275]
xantphos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene [0276]
Zn(CN).sub.2 zinc cyanide
Generic Synthesis Schemes
[0277] The compounds of this invention may be made by a variety of
methods, including well-known standard synthetic methods.
Illustrative general synthetic methods are set out below and then
specific compounds of the invention are prepared in the working
examples. The skilled artisan will appreciate that if a substituent
described herein is not compatible with the synthetic methods
described herein, the substituent may be protected with a suitable
protecting group that is stable to the reaction conditions. The
protecting group may be removed at a suitable point in the reaction
sequence to provide a desired intermediate or target compound. In
all of the schemes described below, protecting groups for sensitive
or reactive groups are employed where necessary in accordance with
general principles of synthetic chemistry. Protecting groups are
manipulated according to standard methods of organic synthesis (T.
W. Green and P. G. M. Wuts, (1991) Protecting Groups in Organic
Synthesis, John Wiley & Sons, incorporated by reference with
regard to protecting groups). These groups are removed at a
convenient stage of the compound synthesis using methods that are
readily apparent to those skilled in the art. The selection of
processes as well as the reaction conditions and order of their
execution shall be consistent with the preparation of compounds of
the present invention. Starting materials are commercially
available or are made from commercially available starting
materials using methods known to those skilled in the art.
[0278] Certain compounds of Formula (I) can be prepared according
to Scheme-10 or analogous methods. Alkylation of a substituted
6-hydroxybenzo[b]thiophene with an optionally substituted
5-halo-6,7-dihydro-5H-cyclopenta[b]pyridine or an optionally
substituted 3-chloro-2,3-dihydrofuro[2,3-b]pyridine is followed by
saponification of the intermediate ester to afford compounds of
Formula (I).
##STR00036##
[0279] Certain compounds of Formula (I) can be prepared according
to Scheme-11 or analogous methods. A Mitsunobu reaction involving a
substituted 6-hydroxybenzo[b]thiophene and an optionally
substituted 6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol or an
optionally substituted 2,3-dihydrofuro[2,3-b]pyridin-3-ol is
followed by saponification of the intermediate ester to afford
compounds of Formula (I).
##STR00037##
[0280] Certain compounds of Formula (I) can be prepared according
to Scheme-12 or analogous methods. Alkylation of a substituted
6-hydroxybenzo[b]thiophene with
2,5-dihalo-6,7-dihydro-5H-cyclopenta[b]pyridine or
3,6-dihalo-2,3-dihydrofuro[2,3-b]pyridine followed by a
palladium-mediated cyanation provides the nitrile. Saponification
of the intermediate ester followed by hydrolysis of the nitrile
affords compounds of Formula (I).
##STR00038##
EXPERIMENTALS
Intermediates
##STR00039##
[0281] a) (3-Chloro-5-methoxyphenyl)(4-methoxybenzyl)sulfane
##STR00040##
[0283] To a mixture of DIPEA (369 mL, 2113 mmol), xantphos (20.38
g, 35.2 mmol), 1-bromo-3-chloro-5-methoxybenzene (156 g, 704 mmol),
(4-methoxyphenyl)methanethiol (109 g, 704 mmol) in toluene (500 mL)
was added Pd.sub.2(dba).sub.3 (32.2 g, 35.2 mmol) at room
temperature and reaction mixture was refluxed for 5 h. After
cooling, water was added to the mixture and extracted with EtOAc.
The organic layer was separated, washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by silica gel column chromatography (EtOAc/hexane) to give
the title compound (yield 170.0 g) as a pale yellow liquid. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.25-7.22 (m, 2H), 6.87-6.82 (m,
3H), 6.69-6.67 (m, 2H), 4.08 (s, 2H), 3.78 (s, 3H), 3.73 (s, 3H).
LCMS (ES) m/z 293 [M+H].sup.+.
b) 3-Chloro-5-methoxybenzenethiol
##STR00041##
[0285] TFA (180 ml, 2336 mmol) was added to the solution of
(3-chloro-5-methoxyphenyl)(4-methoxybenzyl)sulfane (180 g, 611
mmol) in anisole (180 mL) at 0.degree. C. The reaction was stirred
at 85.degree. C. for 2 h under nitrogen atmosphere. Reaction
mixture was quenched with 6N NaOH solution and extracted with
EtOAc. The aqueous layer was acidified with 2N HCl and extracted
with EtOAc. The EtOAc layer was washed with water and brine, dried
over anhydrous Na.sub.2SO.sub.4, filtered and the filtrate was
evaporated under reduced pressure to get crude residue. The residue
was purified by silica gel column chromatography (EtOAc/hexane) to
give the title compound (yield 80.0 g) as pale yellow liquid. LCMS
(ES) m/z 172.88 [M+H]+.
c) Ethyl 4-((3-chloro-5-methoxyphenyl)thio)-3-oxobutanoate
##STR00042##
[0287] To a mixture of 3-chloro-5-methoxybenzenethiol (80 g, 458
mmol) and dry DMF (500 mL) were added K.sub.2CO.sub.3 (63.3 g, 458
mmol) and ethyl 4-chloro-3-oxobutanoate (75 g, 458 mmol) at
0.degree. C. The mixture was stirred at room temperature for 3 h.
The mixture was diluted with water and extracted with EtOAc. The
organic layer was washed successively with water and brine, dried
over MgSO.sub.4, and concentrated in vacuo to get crude product
(100 g). This was used for the next step without any further
purification.
d) Ethyl 2-(4-chloro-6-methoxybenzo[b]thiophen-3-yl)acetate and
Ethyl 2-(6-chloro-4-methoxybenzo[b]thiophen-3-yl)acetate
##STR00043##
[0289] To ethyl 4-((3-chloro-5-methoxyphenyl)thio)-3-oxobutanoate
(100 g, 330 mmol) was added methanesulfonic acid (500 mL) at
0.degree. C. The mixture was stirred at 0.degree. C. under nitrogen
atmosphere for 15 min. The mixture was poured into water and
extracted with EtOAc. The organic layer was washed with brine,
dried over MgSO.sub.4, and concentrated in vacuo. The residue was
purified by silica gel column chromatography (EtOAc/hexane) to give
the mixture of isomers (60 g, ratio 2.5:1) as colorless oil and
used for the next step. LCMS (ES) m/z 285.16 [M+H].sup.+.
e) Ethyl 2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate and
ethyl 2-(6-chloro-4-hydroxybenzo[b]thiophen-3-yl)acetate
##STR00044##
[0291] To a solution of ethyl
2-(4-chloro-6-methoxybenzo[b]thiophen-3-yl)acetate (60 g, 211 mmol)
in DCM (500 mL) was added BBr.sub.3 (29.9 mL, 316 mmol) at
0.degree. C. The mixture was warmed to room temperature and
continued stirring for 6 h at RT. The mixture was quenched with
water and NaHCO.sub.3 solution, and extracted with EtOAc. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4,
and concentrated in vacuo. The residue was purified by silica gel
column chromatography (EtOAc/hexane) to give 27 g of ethyl
2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate) as white solid
and 5.8 g of ethyl
2-(6-chloro-4-hydroxybenzo[b]thiophen-3-yl)acetate).
[0292] Ethyl 2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate
(Desired Compound): .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.07
(s, 1H), 6.95 (d, J=2.4 Hz, 1H), 6.77 (d, J=2.4 Hz, 1H), 5.72 (s,
1H), 4.24 (q, J=9.2 Hz, 2H), 4.08 (s, 2H), 1.30 (t, J=6.9 Hz, 3H).
LCMS (ES) m/z 271.12 (M+H).sup.+.
[0293] Ethyl 2-(6-chloro-4-hydroxybenzo[b]thiophen-3-yl)acetate
(regioisomer Compound): .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
7.07 (s, 1H), 7.01 (d, J=0.9 Hz, 1H), 6.81 (d, J=1.2 Hz, 1H), 5.45
(s, 1H), 4.23 (q, J=7.2 Hz, 2H), 4.08 (s, 2H), 1.29 (t, J=7.2 Hz,
3H). LCMS (ES) m/z 270.93 (M+H).sup.+.
##STR00045##
a) 1-Bromo-2,5-dichloro-3-methoxybenzene
##STR00046##
[0295] To a solution of potassium KO.sup.tBu (20.7 g, 185 mmol))
suspended in toluene (270 mL) and DMPU (90 mL, 746 mmol) was added
methanol (30 mL). The mixture was placed in an oil bath at
80.degree. C. under N.sub.2 with a reflux condenser for 25 minutes
to obtain a solution. The solution was then allowed to cool to room
temperature under N.sub.2, after which
1-bromo-2,5-dichloro-3-fluorobenzene (15 g, 61.5 mmol) was added
dropwise to the solution and the resulting suspension was placed in
an oil bath at 80.degree. C. under N.sub.2. After 4 h, the reaction
mixture was allowed to cool to room temperature and was then
diluted with hexanes (200 mL) and water. The layers were separated
and the aqueous layer was extracted with hexanes. The combined
organic portions were washed with water, dried (MgSO.sub.4),
filtered and concentrated to afford crude. The crude was purified
by silica gel chromatography using 30% EtOAc/pet ether as an eluent
to afford 1-bromo-2,5-dichloro-3-methoxybenzene (13 g, 81% yield)
as off white solid. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.26
(d, J=2.8 Hz, 1H), 6.87 (d, J=2.8 Hz, 1H), 3.93 (s, 3H).
b) (2,5-dichloro-3-methoxyphenyl)(4-methoxybenzyl)sulfane
##STR00047##
[0297] To an argon purged solution of
1-bromo-2,5-dichloro-3-methoxybenzene (13 g, 50.8 mmol),
(4-methoxyphenyl)methanethiol (9.40 g, 61.0 mmol) and DIPEA (17.74
mL, 102 mmol) in toluene (200 mL), xantphos (2.94 g, 5.08 mmol) and
Pd.sub.2(dba).sub.3 (4.65 g, 5.08 mmol) were added at ambient
temperature and heated to 90.degree. C. for 4 h under argon
atmosphere. After 4 h the reaction mixture was cooled to RT and
passed through a pad of Celite.RTM. and the filtrate was diluted
with water and extracted with EtOAc. The organic layer was washed
with water, brine, dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to afford crude. The crude was purified by
silica gel chromatography using 10% EtOAc/pet ether as an eluent to
afford (2,5-dichloro-3-methoxyphenyl)(4-methoxybenzyl)sulfane (13
g, 61% yield) as a yellow solid. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.30-7.25 (m, 2H), 6.87-6.84 (m, 3H), 6.73-6.72 (m, 1H),
4.10 (s, 2H). 3.87 (s, 3H), 3.79 (s, 3H).
c) 2,5-Dichloro-3-methoxybenzenethiol
##STR00048##
[0299] To a stirred solution of
(2,5-dichloro-3-methoxyphenyl)(4-methoxybenzyl)sulfane (13 g, 39.5
mmol) in anisole (70 mL) was added TFA (70 mL, 909 mmol) at ambient
temperature and heated to 100.degree. C. for 2 h. After 2 h the
reaction mixture was diluted with water and extracted with EtOAc.
The organic layer was washed with 2N NaOH solution. The aqueous
layer was washed twice with EtOAc, finally acidified with conc. HCl
and extracted with EtOAc. The organic layer washed with water and
evaporation to afford 2,5-dichloro-3-methoxybenzenethiol (5.5 g,
54.1% yield) as a yellow liquid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 7.28 (s, 1H), 6.99 (s, 1H), 6.02 (brs, 1H).
3.86 (s, 3H).
d) Ethyl 4-((2,5-dichloro-3-methoxyphenyl)thio)-3-oxobutanoate
##STR00049##
[0301] To an ice cooled solution of
2,5-dichloro-3-methoxybenzenethiol (5.5 g, 26.3 mmol) and
K.sub.2CO.sub.3 (10.91 g, 79 mmol) in DMF (50 mL) was slowly added
ethyl 4-chloro-3-oxobutanoate (8.66 g, 52.6 mmol) and the reaction
mass was allowed to stir at ambient temperature. After 2 h the
reaction mixture was diluted with water and extracted with EtOAc.
The organic layer was washed with water, brine, dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to afford
crude which was used as such for next step.
e) Ethyl 2-(4,7-dichloro-6-methoxybenzo[b]thiophen-3-yl)acetate
##STR00050##
[0303] To the above crude ethyl
4-((2,5-dichloro-3-methoxyphenyl)thio)-3-oxobutanoate (6 g, 17
mmol), methane sulfonic acid (5 mL, 77 mmol) was added and stirred
at ambient temperature for 1 h. The reaction mixture was diluted
with water and extracted with EtOAc. The organic layer was washed
with water, brine, dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to afford crude. The crude was purified by
silica gel chromatography using 30% EtOAc/pet-ether as an eluent to
afford ethyl 2-(4,7-dichloro-6-methoxybenzo[b]thiophen-3-yl)acetate
(4 g, 47.2% yield) as an off white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.19 (s, 1H), 7.13 (s, 1H), 4.25-4.15 (q, J=4
Hz, 2H), 4.09 (s, 2H), 3.98 (s, 3H), 1.30 (t, J=4.5 Hz, 3H). LCMS
(ES) m/z 318.8 (M+H).sup.+
f) Ethyl 2-(4,7-dichloro-6-hydroxybenzo[b]thiophen-3-yl)acetate
##STR00051##
[0305] To a stirred solution of ethyl
2-(4,7-dichloro-6-methoxybenzo[b]thiophen-3-yl)acetate (1 g, 3.13
mmol) in DCM (10 mL) was slowly added boron trifluoride methyl
sulfide complex (5 mL, 3.13 mmol) at ambient temperature and
allowed to stir for 12 h. The reaction mixture was diluted with
water and basified with saturated NaHCO.sub.3, extracted with
EtOAc. The organic layer was washed with water, brine, dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to afford
crude. The crude was triturated with ethers to afford ethyl
2-(4,7-dichloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (800 mg,
73.6% yield) as off-white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 10.85 (s, 1H), 7.51 (s, 1H), 7.10 (s, 1H),
4.12-4.05 (q, J=4 Hz, 2H), 3.96 (s, 2H), 1.19 (t, J=4.0 Hz,
3H).
##STR00052##
a) (5-Chloro-2-fluoro-3-methoxyphenyl)(methyl)sulfane
##STR00053##
[0307] To a stirred solution of 4-chloro-1-fluoro-2-methoxybenzene
(8.0 g, 49.8 mmol) in THF (150 mL) was added dropwise (over a
period of 20 min) sec-butyllithium (80 mL, 112 mmol) at -78.degree.
C. and stirred for 30 min. Dimethyl disulfide (9.74 mL, 110 mmol)
was added to the reaction mixture at same temperature. The reaction
mixture was stirred at -78.degree. C. under argon atmosphere for
1.5 h. The reaction mixture was quenched with sat NH.sub.4Cl
solution and partitioned between water and EtOAc. The EtOAc layer
was washed with water and brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and the filtrate was evaporated under
reduced pressure to get crude material. The resultant residue was
purified by column chromatography (100-200 silica mesh and eluent
was 2% EtOAc in pet ether) to afford
(5-chloro-2-fluoro-3-methoxyphenyl)(methyl)sulfane (5.0 g, 48.6%
yield) as an off-white solid. .sup.1H NMR (500 MHz, CDCl.sub.3):
.delta. 6.79-6.76 (m, 2H), 3.86 (s, 3H), 2.36 (s, 3H).
b) 5-Chloro-2-fluoro-1-methoxy-3-(methylsulfinyl)benzene
##STR00054##
[0309] To a stirred solution of
(5-chloro-2-fluoro-3-methoxyphenyl)(methyl)sulfane (5.0 g, 24.19
mmol) in methanol (200 mL) and water (40 mL) was added sodium
periodate (7.76 g, 36.3 mmol) at 0.degree. C. The reaction mixture
was stirred at 26.degree. C. for 16 h. The reaction mixture was
evaporated under reduced pressure the residue was partitioned
between water and EtOAc. The organic layer was washed with water
and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and the
filtrate was evaporated under reduced pressure to get crude
material.
[0310] The resultant residue was purified by column chromatography
(100-200 silica mesh and eluent was 15% EtOAc in pet ether) to
afford 5-chloro-2-fluoro-1-methoxy-3-(methylsulfinyl)benzene (4.0
g, 74.3% yield) as an off-white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. 7.39-7.36 (m, 1H), 7.06 (dd, J=2.5, 7.5 Hz,
1H), 3.93 (s, 3H), 2.83 (s, 3H). LCMS (ES) m/z 223.16
(M+H).sup.+
c) 5-Chloro-2-fluoro-3-methoxybenzenethiol
##STR00055##
[0312] To a stirred solution of
5-chloro-2-fluoro-1-methoxy-3-(methylsulfinyl)benzene (1.50 g, 6.74
mmol) in acetonitrile (60 mL) was added trifluoroacetic anhydride
(1.9 mL, 13.47 mmol) at 0.degree. C. and stirred at same
temperature for 1 h. Then the reaction mixture was stirred at RT
for 1 h. The reaction mixture was concentrated. The residue was
dissolved in a mixture of methanol (10 mL) and TEA (10.0 mL) at
0.degree. C. and stirred for 10 min and concentrated in vacuum. The
mixture was diluted with sat NH.sub.4Cl and extracted with EtOAc.
The organic layer was washed with 1N NaOH. The aqueous layer was
acidified with 1N HCl and extracted with EtOAc. The organic layer
washed with brine dried over anhydrous Na.sub.2SO.sub.4, filtered
and the filtrate was evaporated under reduced pressure to get crude
material. The resulted residue was purified by column
chromatography (100-200 silica mesh, eluent was 10% EtOAc in pet
ether) to obtain 5-chloro-2-fluoro-3-methoxybenzenethiol (0.75 g,
57.8% yield) as an off-white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 6.84 (m, 1H), 6.74 (dd, J=2.4, 6.8 Hz, 1H),
3.88 (s, 3H), 3.81 (s, H)
d) Ethyl
4-((5-chloro-2-fluoro-3-methoxyphenyl)thio)-3-oxobutanoate
##STR00056##
[0314] To the stirred suspension of
5-chloro-2-fluoro-3-methoxybenzenethiol (750 mg, 3.89 mmol) and
potassium carbonate (538 mg, 3.89 mmol) in DMF (10 mL) was added
ethyl 4-chloro-3-oxobutanoate (705 mg, 4.28 mmol) at 0.degree. C.
The reaction mixture was stirred at RT for 2 h. The reaction
mixture was diluted with water and extracted with EtOAc. The
organic layer was separated and dried over anhydrous
Na.sub.2SO.sub.4, filtered and the filtrate was evaporated under
reduced pressure to get crude product ethyl
4-((5-chloro-2-fluoro-3-methoxyphenyl)thio)-3-oxobutanoate (750 mg,
60.1% yield) as a brown liquid. .sup.1H NMR 400 MHz, CDCl.sub.3):
.delta. 6.94-6.92 (m, 1H) 6.86 (dd, J=2.4, 7.2 Hz, 1H), 4.23 (q,
2H), 3.87 (s, 3H), 3.82 (s, 2H), 3.64 (s, 2H), 1.27 (t, J=2.4 Hz,
3H).
e) Ethyl
2-(4-chloro-7-fluoro-6-methoxybenzo[b]thiophen-3-yl)acetate
##STR00057##
[0316] To a stirred solution of ethyl
4-((5-chloro-2-fluoro-3-methoxyphenyl)thio)-3-oxobutanoate (750 mg,
2.338 mmol) was added methanesulfonic acid (3.0 ml, 46.2 mmol) at
0.degree. C. and mixture was stirred at RT for 1 h. The reaction
mixture was partitioned between EtOAc and water, the separated
organic layer was washed with brine solution, dried over anhydrous
Na.sub.2SO.sub.4, filtered and filtrate was evaporated under
reduced pressure to get crude material. The resultant residue was
purified by column chromatography (100-200 silica mesh and eluent
was 15% EtOAc in pet ether) to afford ethyl
2-(4-chloro-7-fluoro-6-methoxybenzo[b]thiophen-3-yl)acetate (450
mg, 58.5% yield) as a pale yellow liquid. .sup.1H NMR 500 MHz,
CDCl.sub.3): .delta. 7.16 (s, 1H) 7.07 (d, J=7.0 Hz, 1H), 4.17 (q,
2H), 4.07 (s, 2H), 3.95 (s, 3H), 1.26 (t, J=5.6 Hz, 3H). LCMS (ES)
m/z 303.25 (M+H).sup.+.
f) Ethyl
2-(4-chloro-7-fluoro-6-hydroxybenzo[b]thiophen-3-yl)acetate
##STR00058##
[0318] To a solution of ethyl
2-(4-chloro-7-fluoro-6-methoxybenzo[b]thiophen-3-yl)acetate (350
mg, 1.156 mmol) in DCM (10 mL) was added BBr.sub.3 (0.164 mL, 1.734
mmol) at -50.degree. C. The reaction mixture was cool to room
temperature for 1 h under nitrogen atmosphere. The Reaction mixture
was quenched with saturated NaHCO.sub.3 solution and partitioned
between water and DCM. The DCM layer was washed with water and
brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and the
filtrate was evaporated under reduced pressure to get crude
material. The resulted residue was purified by column
chromatography (100-200 silica mesh, eluent was 10% EtOAc in pet
ether) to obtain ethyl
2-(4-chloro-7-fluoro-6-hydroxybenzo[b]thiophen-3-yl)acetate (300
mg, 80% yield) as an off-white solid. .sup.1H NMR 500 MHz,
CDCl.sub.3): .delta. 7.15 (s, 1H) 7.05 (d, J=7.0 Hz, 1H), 4.2 (q,
J=5.6 Hz, 2H), 4.08 (s, 2H), 1.26 (t, J=5.6 Hz, 3H). LCMS (ES) m/z
289.19 (M+H).sup.+.
##STR00059##
a) Methyl 2-bromo-6-methylnicotinate
##STR00060##
[0320] Phosphorus oxybromide (21.53 g, 75 mmol) was added to the
stirred solution of 2-hydroxy-6-methylnicotinic acid (5 g, 32.7
mmol), pyridine (0.475 mL, 5.88 mmol) in chlorobenzene (100 mL) at
room temperature under nitrogen. The reaction mixture was refluxed
for 1 h and concentrated under vacuum before treating with an
excess of cold methanol. The solution was stirred for an additional
1 h and again concentrated under vacuum. The residue was dissolved
in water and pH was adjusted to .about.8.0 by adding
K.sub.2CO.sub.3 before extraction of the product with
CH.sub.2Cl.sub.2. The organic layer was washed with water and brine
solution, dried over anhydrous Na.sub.2SO.sub.4. Filtrate was
evaporated completely under reduced pressure to give crude residue.
The resulted crude compound was purified by flash column
chromatography (100-200 silica mesh, eluent was 30% EtOAc in pet
ether) to obtained methyl 2-bromo-6-methylnicotinate (6.1 g, 79%
yield) as colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
8.00 (d, J=7.6 Hz, 1H), 7.18 (d, J=8.0 Hz, 1H), 3.94 (s, 3H), 2.59
(s, 3H); LCMS (ES) m/z 230.0 (M+H).sup.+
b) Methyl
(E)-2-(3-methoxy-3-oxoprop-1-en-1-yl)-6-methylnicotinate
##STR00061##
[0322] Na.sub.2CO.sub.3 (8.43 g, 80 mmol) was added to a solution
of methyl 2-bromo-6-methylnicotinate (6.1 g, 26.5 mmol) and methyl
acrylate (6.08 mL, 67.1 mmol) in mixture of DMA (16.99 mL, 181
mmol) and toluene (55 mL) at room temperature. Then the reaction
mixture was degassed for 15 min. Tri-o-tolylphosphine (0.807 g,
2.65 mmol) and allylpalladium chloride dimer (0.4850 g, 1.326 mmol)
were added and the reaction mixture was stirred at 115.degree. C.
in sealed tube for 5 h. Filtered through pad of Celite.RTM., and
the filtrate was concentrated under reduced pressure. The resultant
crude compound was purified by flash column chromatography on
100-200 mesh silica gel using 20% EtOAc/pet-ether as an eluent to
obtained (E)-methyl
2-(3-methoxy-3-oxoprop-1-en-1-yl)-6-methylnicotinate (3.30 g, 43.0%
yield) as a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 8.53 (dd, J=1.2, 15.2 Hz, 1H), 8.22 (d, J=6.8 Hz, 1H),
7.20-7.11 (m, 2H), 3.94 (s, 3H), 3.82 (s, 3H), 2.60 (s, 3H). LCMS
(ES) m/z 236.09 (M+H).sup.+
c) Methyl 2-(3-methoxy-3-oxopropyl)-6-methylnicotinate
##STR00062##
[0324] 10% Pd--C (300 mg, 2.82 mmol) was added to a solution of
(E)-methyl 2-(3-methoxy-3-oxoprop-1-en-1-yl)-6-methylnicotinate
(3.30 g, 14.03 mmol) in methanol (120 mL) at 25.degree. C. The
reaction mixture was stirred for 3 h at 25.degree. C. under
hydrogen atmospheric pressure of 50 psi. The reaction mixture was
filtered and filtrate was evaporated under pressure to get methyl
2-(3-methoxy-3-oxopropyl)-6-methylnicotinate (2.8 g, 74.3% yield)
as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.08
(d, J=8.0 Hz, 1H), 7.06 (d, J=7.6 Hz, 1H), 3.90 (s, 3H), 3.67 (s,
3H), 3.50 (t, J=7.6 Hz, 2H), 2.81 (t, J=8.0 Hz, 2H), 2.55 (s, 3H).
LCMS (ES) m/z 238.10 (M+H).sup.+
d) 2-Methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one
##STR00063##
[0326] Sodium methoxide (0.956 g, 17.70 mmol) was added to a
solution of methyl 2-(3-methoxy-3-oxopropyl)-6-methylnicotinate
(2.8 g, 11.80 mmol) in THF (30 mL) under nitrogen atmosphere. The
reaction mixture was warmed to reflux during 2 h. The solvent was
removed under vacuo and HCl (20 ml, 90 mmol) 4.5 M was added, the
mixture was stirred 2 h at reflux. The reaction mixture was
dissolved in water and pH was adjusted to .about.8.0 by adding
K.sub.2CO.sub.3 before extraction of the product with
CH.sub.2Cl.sub.2. The organic layer was washed with water and brine
solution, dried over anhydrous Na.sub.2SO.sub.4. Filtrate was
evaporated completely under reduced pressure to give crude residue.
The crude residue was purified by silica gel column chromatography
by using EtOAc in hexane as eluent, the product was eluted at 40%
EtOAc/Pet-ether to get
2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (1.3 g, 66.1%
yield) as a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 7.91 (d, J=8.4 Hz, 1H), 7.19 (d, J=7.6 Hz, 1H), 3.24 (t,
J=6.0 Hz, 2H), 2.78 (t, J=8.0 Hz, 2H), 2.67 (s, 3H). LCMS (ES) m/z
147.98 (M+H).sup.+
e) 2-Methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol
##STR00064##
[0328] NaBH.sub.4 (0.334 g, 8.83 mmol) was added lot wise to the
stirred solution of
2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (1.3 g, 8.83
mmol) in methanol (30 ml) at 0.degree. C. and the mixture was
stirred at 0.degree. C. for 2 h. The reaction mixture was diluted
with water and mixture was concentrated under reduced pressure. The
resulted residue was partitioned between EtOAc and water, the
separated organic layer was washed with brine solution, dried over
anhydrous Na.sub.2SO.sub.4, filtered and filtrate was evaporated
under reduced pressure. The resulted crude compound was purified by
flash column chromatography (100-200 silica mesh, eluent was 70%
EtOAc in pet ether) to obtained
2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (0.900 g, 67.6%
yield) as a colorless oil. .sup.1H NMR (300 MHz, CDCl.sub.3):
.delta. 7.59 (d, J=7.5 Hz, 1H), 7.01 (d, J=7.5 Hz, 1H), 5.25 (s,
1H), 3.18-3.08 (m, 1H), 2.95-2.84 (m, 1H), 2.59 (s, 4H), 2.04-1.94
(m, 2H). LCMS (ES) m/z 150.3 [M+H].sup.+
f) 5-Chloro-2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridine
##STR00065##
[0330] To solution of
2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (20.66 g, 139
mmol) in DCM (200 mL) was added thionyl chloride (6.74 mL, 92 mmol)
at RT and stirred for 20 min, solvents were evaporated under
reduced pressure to afford crude product. The crude product was
used for the next step without further purification.
##STR00066##
a) 6,7-Dihydro-5H-cyclopenta[b]pyridin-5-one
##STR00067##
[0332] KMnO.sub.4 (53.0 g, 336 mmol) dissolved in water (2000 mL)
was added to the stirred solution of
6,7-dihydro-5H-cyclopenta[b]pyridine (20 g, 168 mmol) and
MgSO.sub.4.cndot.7H.sub.2O (40.4 g, 336 mmol) in tert-butanol (500
mL) at 25.degree. C. and the reaction mixture was stirred at
30.degree. C. for 3 h. The reaction mixture was filtered through a
Celite.RTM. bed, partitioned between EtOAc and water, the separated
organic layer was washed with brine solution, dried over anhydrous
Na.sub.2SO.sub.4, filtered and filtrate was concentrated under
reduced pressure. The resulted crude compound was purified by flash
column chromatography on 100-200 silica gel, using 20-30% EtOAc-Pet
ether as an eluent to obtained
6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (10 g, 44.7% yield) as an
off white solid. LCMS (ES) m/z 134.01 [M+H].sup.+.
b) 6,7-Dihydro-5H-cyclopenta[b]pyridin-5-ol
##STR00068##
[0334] NaBH.sub.4 (25.6 g, 676 mmol) was added portion wise to the
stirred solution of 6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (60
g, 451 mmol) in methanol (600 mL) at 0.degree. C. and the reaction
mixture was stirred at 25.degree. C. for 1 h under nitrogen
atmosphere. The reaction mixture was quenched with water and then
solvent was distilled off. The residue was partitioned between
EtOAc and water, the separated organic layer was washed with brine
solution, dried over anhydrous Na.sub.2SO.sub.4, filtered and
filtrate was concentrated under reduced pressure. The resulted
crude compound was purified by flash column chromatography on
100-200 silica gel, using EtOAc-Pet ether as an eluent to obtained
6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (40 g, 62.5% yield) as an
off white solid. LCMS (ES) m/z 136.11 [M+H].sup.+.
c) 5-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridine
##STR00069##
[0336] To a solution of 6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (5
g, 37.0 mmol) in DCM (50 mL) was added thionyl chloride (4.05 mL,
55.5 mmol) at 0.degree. C. and the reaction mixture was stirred at
25.degree. C. for 1 h under nitrogen atmosphere. Reaction mixture
was concentrated under reduced pressure to get crude product
5-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (5.5 g) as a brown
gummy liquid. Crude was used for the next step without further
purification. LCMS (ES) m/z 154.19 [M+H].sup.+.
##STR00070##
a) Ethyl
(E)-1-(3-amino-3-oxoprop-1-en-1-yl)-2-oxocyclopentane-1-carboxyl-
ate
##STR00071##
[0338] Ethyl 2-oxocyclopentanecarboxylate (150 g, 960 mmol) was
added to the stirred solution of propiolamide (113 g, 1633 mmol)
and Na.sub.2CO.sub.3 (112 g, 1056 mmol) in water (1500 mL) at
0.degree. C. and the mixture was stirred at RT for 6 h. The
reaction mixture was diluted with water and extracted with DCM, the
organic layer was washed with brine solution, dried over anhydrous
Na.sub.2SO.sub.4, filtered and filtrate was evaporated under
reduced pressure to get crude product. The resulted crude compound
was purified by flash column chromatography (100-200 silica mesh,
eluent was 80% EtOAc in pet ether) to obtained (E)-ethyl
1-(3-amino-3-oxoprop-1-en-1-yl)-2-oxocyclopentanecarboxylate (150
g, 68.8% yield) as an off-white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): .delta. 6.58 (d, J=10.4 Hz, 1H), 5.97 (dd, J=2.0, 10.0
Hz, 1H), 5.89 (brs, 2H), 4.26 (q, J=4.8 Hz, 2H), 2.50-2.42 (m, 1H),
2.26-2.19 (m, 1H), 2.08-1.88 (m, 3H), 1.76-1.72 (m, 1H), 1.31 (t,
J=7.2 Hz, 3H). LCMS (ES) m/z 226.23 [M+H].sup.+
b) 6,7-Dihydro-5H-cyclopenta[b]pyridin-2-ol
##STR00072##
[0340] Conc. HCl (209 ml, 6882 mmol) was added to the (E)-ethyl
1-(3-amino-3-oxoprop-1-en-1-yl)-2-oxocyclopentanecarboxylate (155
g, 688 mmol) at room temperature and the mixture was stirred at
130.degree. C. for 5 h. The reaction mixture was concentrated and
poured into ice. The pH was adjusted to .about.7.0 by dropwise
addition of saturated aqueous NaHCO.sub.3 solution, and filtered
the precipitated solid. The solid was washed with water to get
6,7-dihydro-5H-cyclopenta[b]pyridin-2-ol (80 g, 86% yield) as an
off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 7.71
(s, 1H), 6.62 (s, 1H), 2.89-2.85 (m, 2H), 2.75-2.72 (m, 2H),
2.11-2.04 (m, 2H). LCMS (ES) m/z 136.07 [M+H].sup.+
c) 2-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridine
##STR00073##
[0342] A mixture of 6,7-dihydro-5H-cyclopenta[b]pyridin-2-ol (70 g,
518 mmol), POCl.sub.3 (200.0 ml, 2146 mmol) and DMF (10 mL) was
stirred under nitrogen atmosphere at 120.degree. C. for 3 h. After
cooling, the mixture was poured into ice water, basified with 8 M
NaOH aqueous solution and extracted with AcOEt. The extract was
washed with brine, dried over anhydrous Na.sub.2SO.sub.4, and
concentrated. The resulted crude compound was purified by flash
column chromatography (100-200 silica mesh, eluent was 10% EtOAc in
pet ether) to obtained
2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (35 g, 43.7% yield)
as an off white solid. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
7.44 (d, J=7.6 Hz, 1H), 7.06 (d, J=8.0 Hz, 1H), 3.01 (t, J=7.2 Hz,
2H), 2.92 (t, J=7.2 Hz, 2H), 2.18-2.10 (m, 2H). LCMS (ES) m/z
154.09 [M+H].sup.+
d) 2-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one
##STR00074##
[0344] KMnO.sub.4 (72.0 g, 456 mmol) dissolved in water (3.5 L) was
added to the stirred solution of
2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (35.00 g, 228 mmol)
and magnesium sulfate heptahydrate (68.2 g, 456 mmol) in
tert-butanol (875 mL) and the reaction mixture was stirred at RT
for 2 h under nitrogen atmosphere. The reaction mixture was
filtered through a Celite.RTM. pad, partitioned between EtOAc and
water. The separated organic layer was washed with brine solution,
dried over anhydrous Na.sub.2SO.sub.4, filtered and filtrate was
concentrated under reduced pressure. The resulted crude compound
was purified by flash column chromatography on 100-200 silica gel,
using 20-30% EtOAc-Pet ether as an eluent to obtained
2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (25 g, 65.4%
yield) as an off white solid. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 7.97 (d, J=8.4 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 3.28-3.25
(m, 2H), 2.82-2.79 (m, 2H); LCMS (ES) m/z 168.08 [M+H].sup.+
e) 2-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol
##STR00075##
[0346] Sodium borohydride (108 mg, 2.86 mmol) was added lot wise to
the stirred solution of
2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (480 mg, 2.86
mmol) in methanol (100 mL) at 0.degree. C. and the mixture was
stirred at 0.degree. C. for 1 h. The reaction mixture was diluted
with water and mixture was concentrated under reduced pressure. The
resulted residue was partitioned between EtOAc and water, the
separated organic layer was washed with brine solution, dried over
anhydrous Na.sub.2SO.sub.4, filtered and filtrate was evaporated
under reduced pressure. The resulted crude compound was purified by
flash column chromatography (100-200 silica mesh, eluent was 30%
EtOAc in pet ether) to obtained
2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (450 mg, 91%
yield) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3):
.delta. 7.67 (d, J=8.0 Hz, 1H), 7.19 (d, J=8.0 Hz, 1H), 5.28-5.26
(m, 1H), 3.13-3.10 (m, 1H), 2.95-2.92 (m, 1H), 2.05-2.02 (m, 1H)
1.89 (m, 1H). LCMS (ES) m/z 170.16 [M+H].sup.+
f) 2,5-Dichloro-6,7-dihydro-5H-cyclopenta[b]pyridine
##STR00076##
[0348] SOCl.sub.2 (0.232 ml, 3.18 mmol) was added to the stirred
solution of 2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol
(450.00 mg, 2.65 mmol) in DCM (50 ml) and the mixture was stirred
at room temperature for 3 h. The reaction mixture was concentrated
under reduced pressure and crude used for the next step. LCMS (ES)
m/z 188.15 [M+H].sup.+.
##STR00077## ##STR00078##
a) Butoxy-1,1,1-trifluorobut-3-en-2-one
##STR00079##
[0350] To a stirred solution of 1-(vinyloxy)butane (50 g, 499
mmol), pyridine (40.4 mL, 499 mmol) in Chloroform (500 mL) at
0.degree. C. 1,1,1,5,5,5-hexafluoropentane-2,4-dione (104 g, 499
mmol) in chloroform (200 ml) was added and stirred for 16 h After
completion of reaction, mixture was poured into cool water. The
solution was extracted by DCM and washed with water followed by
brine. The organic layer was dried over anhydrous sodium sulphate
and solvent was removed under reduced pressure. The crude was
purified by flash column chromatography on silica gel (100-200
mesh), eluting with 0-30% gradient of EtOAc in hexane to afford
(E)-1-ethoxy-5,5,5-trifluoropent-1-en-3-one (70 g, 73% yield) as a
liquid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.90 (d, J=12 Hz,
1H), 5.86 (d, J=2.4 Hz, 1H), 4.03 (t, J=6.4, 2H), 1.77-1.70 (m,
2H), 1.48-1.39 (m, 2H), 0.95 (t, J=7.6 Hz, 3H).
b) 2-Oxo-6-(trifluoromethyl)-1,2-dihydropyridine-3-carboxamide
##STR00080##
[0352] To a stirred solution of malonamide (39.2 g, 384 mmol) in
methanol (300 mL) at 0.degree. C.
(E)-1-ethoxy-5,5,5-trifluoropent-1-en-3-one (70 g, 384 mmol) in
methanol (300 mL) was added and reaction mixture was stirred at
reflux temperature for 6 h, After completion of reaction mixture
was concentrated, poured into cool water and acidified with dil.HCl
(pH 2) to get solid. Solid was filtered and dried to get pure
compound
2-oxo-6-(trifluoromethyl)-1,2-dihydropyridine-3-carboxamide (60 g,
73.1% yield) as off white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 13.66 (brs, 1H), 8.46 (brs, 2H), 8.07
(brs, 1H), 7.38 (brs, 1H). LCMS (ES.sup.+) m/z 207.11
[M+H].sup.+.
c) 2-Oxo-6-(trifluoromethyl)-1,2-dihydropyridine-3-carboxylic
acid
##STR00081##
[0354] To a stirred solution of
2-oxo-6-(trifluoromethyl)-1,2-dihydropyridine-3-carboxamide (60 g,
291 mmol) in methanol (300 mL), water (100 mL) LiOH (20.91 g, 873
mmol) was added at room temperature and reaction mixture was
stirred at reflux temperature for 24 h. After completion of the
reaction, mixture was poured into cool water and acidified with 1N
HCl. to get solid. Solid was filtered and dried to get pure
compound 2-oxo-6-(trifluoromethyl)-1,2-dihydropyridine-3-carboxylic
acid (52 g, 86% yield) as off white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 12.74 (brs, 1H), 8.72 (d, J=6.5 Hz, 1H),
7.10 (d, J=5.5 Hz, 1H). LCMS (ES) m/z 208.08 (M+H).sup.+.
d) Methyl 2-bromo-6-(trifluoromethyl)nicotinate
##STR00082##
[0356] To the stirred solution of
2-hydroxy-6-(trifluoromethyl)nicotinic acid (23 g, 111 mmol) and
pyridine (8.98 mL, 111 mmol) in chlorobenzene (250 mL) phosphorus
oxybromide (63.7 g, 222 mmol) was added small portions wise at room
temperature and the mixture was stirred at 120.degree. C. for 16 h.
After completion, the reaction mixture was concentrated under
vacuum. The residue was cooled 0.degree. C. and added excess cold
methanol slowly. The solution stirred additional 1 h and again
concentrated under vacuum. The residue dissolved in water and pH
adjusted to .about.8 using K.sub.2CO.sub.3 before extraction with
EtOAc. The organic layer was separated and dried over anhydrous
Na.sub.2SO.sub.4, filtered and the filtrate was concentrated under
reduced pressure to obtain as a brown liquid. The crude was
purified by flash column chromatography on silica gel (100-200
mesh), eluting with 0-10% gradient of EtOAc in hexane to afford
4-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole in (400 g, 42.1%)
yields. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.20 (d, J=8 Hz,
1H), 7.71 (d, J=8.0 Hz, 1H), 4.01 (s, 3H).
e) Methyl
(E)-2-(3-methoxy-3-oxoprop-1-en-1-yl)-6-(trifluoromethyl)nicotin-
ate
##STR00083##
[0358] To a stirred solution of methyl
2-bromo-6-(trifluoromethyl)nicotinate (21 g, 73.9 mmol), methyl
acrylate (16.75 mL, 185 mmol) and sodium carbonate (23.51 g, 222
mmol) in N,N-dimethylacetamide (DMA) (100 mL), toluene (400 mL),
allylpalladium chloride dimer (1.353 g, 3.70 mmol),
tri-o-tolylphosphine (2.250 g, 7.39 mmol) was added at room
temperature in a sealed tube. The resulting reaction mixture was
stirred for 16 h at 120.degree. C. After completion, the reaction
mixture was filtered through a Celite.RTM. bed and was washed with
EtOAc thoroughly. The filtrate was concentrated to get crude
residue The crude compound was purified by column chromatography
(100-200 mesh silica gel) using 10% EtOAc in pet-ether as an eluent
to get (E)-methyl
2-(3-methoxy-3-oxoprop-1-en-1-yl)-6-(trifluoromethyl)nicotinate (13
g, 60.8% yield) as yellow solid. .sup.1H NMR (400 MHz, DMSO-ds) S
8.45 (d, J=15.6 Hz, 1H), 8.39 (dd, J=0.4, 8.0 Hz, 1H), 7.71 (d,
J=8.4 Hz, 1H), 7.26 (d, J=2.8 Hz, 1H), 4.01 (s, 3H), 3.84 (s, 3H).
LCMS (ES) m/z 290.22 [M+H].sup.+.
f) Methyl
2-(3-methoxy-3-oxopropyl)-6-(trifluoromethyl)nicotinate
##STR00084##
[0360] To a stirred solution of (E)-methyl
2-(3-methoxy-3-oxoprop-1-en-1-yl)-6-(trifluoromethyl)nicotinate
(6.0 g, 20.75 mmol) in ethanol (180 mL), Pd/C (2.65 g) was added at
room temperature. The mixture was stirred at room temperature for 1
h under hydrogen balloon pressure, filtered through pad of
Celite.RTM., and filtrate was concentrated under vacuo to afford
methyl 2-(3-methoxy-3-oxopropyl)-6-(trifluoromethyl)nicotinate (4.5
g, 14.88 mmol, 71.7% yield) as a colorless liquid. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 8.36 (d, J=8.0 Hz, 1H), 7.60 (d, J=8.0 Hz,
1H), 3.99 (s, 3H), 3.70 (s, 3H), 3.60 (t, J=6.5 Hz, 2H), 2.88 (t,
J=7.0 Hz, 2H). LCMS (ES) m/z 292.08 [M+H].sup.+.
g)
2-(Trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one
##STR00085##
[0362] To a stirred solution of methyl
2-(3-methoxy-3-oxopropyl)-6-(trifluoromethyl)nicotinate (8 g, 27.5
mmol) in dry methanol (100 mL), sodium methoxide (2.226 g, 41.2
mmol) was added at room temperature and the mixture was stirred for
8 h at 80.degree. C. under argon. The solvent was removed under
reduced pressure and the resultant residue was dissolved in HCl
(12.50 mL, 411 mmol) and stirred at 80.degree. C. for 8 h. After
completion, the reaction mixture was cooled to 0.degree. C. and
basified with 3N NaOH solution and partitioned between EtOAc and
water. The separated organic layer was washed with brine solution,
dried over anhydrous Na.sub.2SO.sub.4, filtered and filtrate was
evaporated under reduced pressure to get the crude residue
2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (4.0
g, 45.9% yield) as brown colored gum. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.20 (d, J=8.0 Hz, 1H), 7.72 (d, J=8.0 Hz, 1H),
3.39 (t, J=5.0 Hz 2H), 2.88 (t, J=6.0 Hz, 2H); LCMS (ES) m/z 202.27
[M+H].sup.+.
h) 2-(Trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol
##STR00086##
[0364] To a stirred solution of
2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-one (4.0
g, 19.89 mmol) in methanol (50 mL), sodium borohydride (0.752 g,
19.89 mmol) was added lot wise at 0.degree. C. and the mixture was
stirred at room temperature for 2 h. After completion, the reaction
mixture was diluted with water and was concentrated under reduced
pressure. The resulted residue was partitioned between EtOAc and
water. The separated organic layer was washed with brine solution,
dried over anhydrous Na.sub.2SO.sub.4, filtered and filtrate was
evaporated under reduced pressure. The crude obtained was purified
by column chromatography on silica gel (100-200 mesh) eluted with
20-50% gradient of EtOAc in hexanes to afford
2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (3.1
g, 13.84 mmol, 69.6% yield) as an off-white solid. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.86 (d, J=8 Hz, 1H), 7.56 (d, J=7.6 Hz,
1H), 5.36-5.31 (m, 1H), 3.29-3.25 (m, 1H), 3.10-2.90 (m, 1H),
2.7-2.6 (m, 1H), 2.05-1.90 (m, 1H); LCMS (ES) m/z 204.22
[M+H].sup.+.
i)
5-Chloro-2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridine
##STR00087##
[0366] To a stirred solution of
2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (56.3
mg, 0.277 mmol) in DCM (15 mL) was added SOCl.sub.2 (0.020 mL,
0.277 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 30 min. and then evaporated under reduced
pressure to get residue. The crude compound used directly for the
next step.
##STR00088##
a) Ethyl
3-oxo-6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]pyridine-2-carbo-
xylate
##STR00089##
[0368] To a solution of ethyl 2-hydroxyacetate (19.50 g, 187 mmol)
in 1,2-dimethoxyethane (DME) (200 mL) was added NaH (4.49 g, 187
mmol) at 0.degree. C. and then solution of ethyl
2-chloro-6-(trifluoromethyl)nicotinate (19 g, 74.9 mmol) in
1,2-dimethoxyethane (DME) (200 mL) was added to the reaction
mixture at RT. The resulting reaction mixture was stirred at
75.degree. C. for 2 h. The reaction mixture was quenched with
saturated sodium bicarbonate, extracted with EtOAc. The organic
layer was washed successively with water and brine, dried over
MgSO.sub.4, and concentrated in vacuo to get crude. The residue was
purified by silica gel column chromatography (EtOAc/Pet ether) to
afford the title compound (9.6 g) as yellow solid. LCMS (ES) m/z
276.07 [M+H].sup.+.
b) 6-(Trifluoromethyl)furo[2,3-b]pyridin-3(2H)-one
##STR00090##
[0370] To a solution of ethyl
3-oxo-6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]pyridine-2-carboxylate
(4 g, 14.54 mmol) in 1,4-dioxane (40 mL) was added HCl (11.04 mL,
363 mmol) at RT. The reaction mixture was heated to 100.degree. C.
for 24 h. The reaction mixture was quenched with saturated sodium
bicarbonate and extracted with EtOAc. The organic layer was washed
successively with water and brine, dried over MgSO.sub.4, and
concentrated in vacuo to give the title compound (1.5 g) as yellow
solid. The crude compound used for the next step without further
purification. LCMS (ES) m/z 203.78 [M+H].sup.+.
d) 6-(Trifluoromethyl)-2,3-dihydrofuro[2,3-b]pyridin-3-ol
##STR00091##
[0372] The title compound was prepared as a white solid according
to the procedures of Scheme 7, Step h, LCMS (ES) m/z 206.10
[M+H].sup.+.
##STR00092##
a) Ethyl 2-chloronicotinate
##STR00093##
[0374] To a stirred solution of 2-chloronicotinic acid (25 g, 159
mmol) in DMF (300 mL) was added MeI (11.91 mL, 190 mmol),
K.sub.2CO.sub.3 (54.8 g, 397 mmol) at rt. The reaction mixture was
stirred at RT for 3 h. The Reaction mixture was diluted with EtOAc
(500 mL) washed with water (4.times.500 mL) and brine (500 mL).
Organic layer was dried over anhydrous sodium sulphate filtered and
concentrated to afford ethyl 2-chloronicotinate (26 g, yield 95%)
as off white solid. .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
8.53-8.51 (m, 1H), 8.18-15 (m, 1H), 7.34-7.31 (m, 1H), 4.0 (s, 3H);
LCMS (ES) m/z 171.94 [M+H].sup.+.
b) Ethyl 3-oxo-2,3-dihydrofuro[2,3-b]pyridine-2-carboxylate
##STR00094##
[0376] To a stirred suspension of NaH (10.91 g, 455 mmol) in
1,2-dimethoxyethane (1200 mL) was added ethyl 2-hydroxyacetate
(39.4 g, 379 mmol) at 0.degree. C. Reaction mixture was stirred at
RT for 30 min. After that methyl 2-chloronicotinate (26 g, 152
mmol) in DME (150 mL) was added to the reaction mixture and the
resulting mixture was heated at 75.degree. C. for 2 h. Reaction
mixture was concentrated. The crude was basified with saturated
sodium bicarbonate and washed with EtOAc (1.times.500 mL). Aqueous
layer was acidify with acetic acid, extracted with DCM (2.times.500
mL), washed with water (500 mL) and brine (500 mL). Organic layer
was dried over anhydrous sodium sulphate filtered and concentrated.
The crude residue was purified by column chromatography (100-200
mesh silica). Compound Eluted at 12% EtOAc in hexane. The eluents
were concentrated at reduced pressure and to affording ethyl
3-oxo-2,3-dihydrofuro[2,3-b]pyridine-2-carboxylate (16 g, yield
35.2%) as off white solid. LCMS (ES) m/z 207.96 [M+1].sup.+. [0377]
c) Furo[2,3-b]pyridin-3(2H)-one:
##STR00095##
[0378] To a stirred solution of ethyl
3-oxo-2,3-dihydrofuro[2,3-b]pyridine-2-carboxylate (12 g, 57.9
mmol) in HCl (9.65 ml, 57.9 mmol) was stirred at 100.degree. C. for
1 h. Reaction mixture was basify with sat sodium bicarbonate
diluted with EtOAc (500 mL) washed with water (200 mL) and brine
(200 mL). Organic layer was dried anhydrous sodium sulphate
filtered and concentrated. Crude was purified by column
chromatography (100-200 mesh silica) to afford
furo[2,3-b]pyridin-3(2H)-one (8 g, yield 84%) as off white solid.
.sup.1H NMR (500 MHz, CDCl.sub.3): .delta. 8.59-8.58 (m, 1H),
8.06-8.04 (m, 1H), 7.16-7.14 (m, 1H), 4.75 (s, 2H). LCMS (ES) m/z
136.07 [M+H].sup.+.
d) 2,3-Dihydrofuro[2,3-b]pyridin-3-ol
##STR00096##
[0380] To a stirred solution of furo[2,3-b]pyridin-3(2H)-one (8 g,
59.2 mmol) in methanol (80 mL) was added NaBH.sub.4 (2.24 g, 59.2
mmol) at 0.degree. C. Reaction mixture was stirred at RT for 3 h.
The reaction mixture was diluted with EtOAc (200 mL) washed with
water (200 mL) and brine (200 mL). Organic layer was dried
anhydrous sodium sulphate filtered and concentrated. Crude was
purified by column chromatography (100-200 mesh silica gel) and the
compound eluted at 80% EtOAc in hexane. Eluents were concentrated
to affording 2,3-dihydrofuro[2,3-b]pyridin-3-ol (4 g, 47.7%) as off
white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 8.06-8.05
(m, 1H), 7.77-7.75 (m, 1H), 6.95-6.92 (m, 1H), 5.77-5.76 (d, J=6
Hz, 1H), 5.30-5.27 (m, 1H), 4.57-4.54 (m, 1H), 4.24-4.21 (m, 1H).
LCMS (ES) m/z 138.12 [M+H].sup.+.
##STR00097##
Example 1
Preparation of
2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]thio-
phen-3-yl)acetic Acid
a) Ethyl
2-(4-chloro-6-(6,7-dihydro-5H-cyclopenta[b]pyridin-5-yloxy)benzo[-
b]thiophen-3-yl)acetate
##STR00098##
[0382] To a stirred solution of ethyl
2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (5 g, 18.47
mmol) and 5-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (4.26 g,
27.7 mmol) in DMF (50 mL) was added K.sub.2CO.sub.3 (12.76 g, 92
mmol) at RT. The reaction mixture was heated to 80.degree. C. for 1
h. The mixture was diluted with water and extracted with EtOAc. The
organic layer was washed successively with water and brine, dried
over MgSO.sub.4, and concentrated in vacuo to get crude. The
residue was purified by silica gel column chromatography
(EtOAc/hexane) to give the title compound (6.5 g) as brown gummy
liquid. LCMS (ES) m/z 388.17 [M+H].sup.+.
b)
2-(4-Chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]th-
iophen-3-yl)acetic Acid
##STR00099##
[0384] To a stirred solution of ethyl
2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]thio-
phen-3-yl)acetate (40 g, 103 mmol) in THF (200 mL), methanol (200
mL) and water (100 mL) was added lithium hydroxide (monohydrate)
(12.35 g, 516 mmol) at RT and the reaction mixture was stirred at
25.degree. C. for 1 h. The reaction mixture was neutralized with
dilute HCl carefully and the precipitated compound was filtered and
dried under reduced pressure to obtained
2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)ben-
zo[b]thiophen-3-yl)acetic acid (32 g, 86% yield) as an off-white
solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 12.5 (brs, 1H),
8.50 (dd, J=1.2, 4.5 Hz, 1H), 7.78-7.82 (m, 2H), 7.47 (s, 1H),
7.22-7.20 (m, 1H), 7.13 (d, J=2.1 Hz, 1H), 6.00-6.04 (m, 1H), 4.00
(s, 2H), 3.06-3.11 (m, 1H), 2.89-2.99 (m, 1H), 2.62-2.72 (m, 1H),
2.06-2.15 (m, 1H). LCMS (ES) m/z 360.05 [M+H].sup.-. Chiral HPLC:
49.92%:50.08%
Analytical SFC Condition
Column/dimensions: Chiralpak AD-H (250.times.4.6) mm, 5.mu.
% CO2: 60.0%
[0385] % Co solvent: 40.0% (100% MeOH) Total Flow: 3.0 g/min
Back Pressure: 100 bar
Temperature: 30.0.degree. C.
UV: 237 nm
Preparative SFC Condition
Column/dimensions: Chiralpak AD-H (250.times.21) mm, 5.mu.
% CO.sub.2: 65.0%
[0386] % Co solvent: 35.0% (100% Methanol) Total Flow: 60.0
g/min
Back Pressure: 100.0 bar
UV: 284 nm
[0387] Stack time: 8.8 min
Load/Inj: 15.0 mg
[0388] Retention time: Peak 1-3.15 min, Peak 2-3.73 min.
Purity: Peak 1-99.73%, Peak 2-98.00%.
Solubility: Methanol+ACN+DCM+THF
Chiral Separation of Example 1
[0389] Example 1a (First eluted enantiomer):
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid (11 g, 34.3% yield). LCMS (ES) m/z 360.18
[M+H].sup.+. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. 12.4
(brs, 1H), 8.50 (dd, J=1.2, 4.8 Hz, 1H), 7.78-7.83 (m, 2H), 7.49
(s, 1H), 7.23-7.20 (m, 1H), 7.13 (d, J=2.1 Hz, 1H), 6.00-6.04 (m,
1H), 4.01 (s, 2H), 3.06-3.11 (m, 1H), 2.95-2.98 (m, 1H), 2.66-2.68
(m, 1H), 2.09-2.12 (m, 1H).
[0390] Chiral HPLC: 99.73%. Absolute stereochemistry was determined
by vibrational circular dichroism (VCD).
[0391] Example 1b (Second eluted enantiomer):
(R)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic acid (10.3 g, 32.1% yield) as an off white
solids. LCMS (ES) m/z 360.15 (M+H).sup.+. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. 12.4 (brs, 1H), 8.50 (dd, J=1.2, 4.8 Hz,
1H), 7.78-7.83 (m, 2H), 7.49 (s, 1H), 7.23-7.20 (m, 1H), 7.13 (d,
J=2.1 Hz, 1H), 6.00-6.04 (m, 1H), 4.01 (s, 2H), 3.06-3.11 (m, 1H),
2.95-2.98 (m, 1H), 2.66-2.68 (m, 1H), 2.09-2.12 (m, 1H). Chiral
HPLC: 98.00%.
Chiral Synthesis of Example 1a,
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic Acid
##STR00100## ##STR00101##
[0392] a) (3-Chloro-5-methoxyphenyl)(4-methoxybenzyl)sulfane
##STR00102##
[0394] A 250 L reactor was charged with
1-bromo-3-chloro-5-methoxybenzene (7.5 kg, 33864.6 mmol) and
(4-methoxyphenyl)methanethiol (5.74 kg, 37217.3 mmol). Toluene (30
L) was charged to the reaction mass. DIPEA (11.83 L, 67701.9 mmol)
was added at 25.degree. C. slowly into the above reaction mixture.
The reaction mixture was degassed with N.sub.2 for 20 min.
Pd.sub.2(dba).sub.3 (1.55 kg, 1692.66 mmol) and xantphos (0.980 kg,
1692.66 mmol) were added slowly into above reaction mixture (Note:
The reaction mass color changes pale yellow to dark color). The
reaction mixture was again degassed with N.sub.2 for 15 min. The
reaction mass was stirred at 110.degree. C. for 3 h. Completion of
the reaction was monitored by TLC (5% EtOAc in pet ether, R.sub.f
value of the product is 0.5). After completion of reaction, the
reaction mass was cooled to 25.degree. C. and filtered on a
Celite.RTM. bed. The Celite.RTM. bed was washed with EtOAc. DM
water was added to the filtrate and stirred at 25-30.degree. C. for
5-10 min. The combined layers were transferred to a 250 L reactor.
The Aqueous and EtOAc layers were separated. Sodium chloride
solution was added to the EtOAc and stirred at 25-30.degree. C. for
5-10 min. The combined layers were transferred to 250 L reactor.
The aqueous and EtOAc layers were separated. The EtOAc layer was
dried over anhydrous Na.sub.2SO.sub.4 and filtered.
Na.sub.2SO.sub.4 washed with EtOAc. The EtOAc was transferred to a
250 L reactor and evaporated below 40-45.degree. C. under vacuum.
After completion of evaporation, the thick yellow liquid was
subjected to drying by rotary evaporation at 40-45.degree. C. for
1.0 h. drying was terminated and the thick yellow liquid was
obtained (13.5 kg, crude). A chromatography column was packed with
silica gel (20.0 kg, 100-200 mesh). The crude compound dissolved in
DCM and loaded into the column. Run the mobile phase with hexane
(50 L). Then followed by increasing the polarity from 2-5% EtOAc in
hexane (500 L). All pure fractions (by TLC) collected and
concentrated under reduced pressure at 40-45.degree. C. (8.7 kg,
yield 87.17%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.25-7.22
(m, 2H), 6.87-6.82 (m, 3H), 6.69-6.67 (m, 2H), 4.08 (s, 2H), 3.78
(s, 3H), 3.73 (s, 3H).
b) 3-Chloro-5-methoxybenzenethiol
##STR00103##
[0396] A 100 L reactor was charged with a solution of
3-chloro-5-methoxyphenyl(4-methoxybenzyl)sulfane (4.6 kg, 15604.01
mmol) in DCM (46 L). Anisole (15.35 kg, 141945.6 mmol) was added to
the reaction mass and cooled to 0.degree. C.
Trifluoromethanesulfonic acid (1.38 L, 15604 mmol) was added
dropwise to the reaction mass at 0-5.degree. C. for 20 min (Note:
The reaction mass color changed from pale yellow to red color). The
reaction mass was stirred at 25.degree. C. for 16 h under N.sub.2
atmosphere. The reaction was monitored by TLC (5% EtOAc in pet
ether, R.sub.f value of the product is 0.6). After completion of
reaction, the reaction mass was cooled to 0.degree. C. 2N NaOH
solution was added dropwise to the reaction mass at 0-10.degree. C.
until the pH of the reaction mass was .about.13. The resulting
mixture stirred at 25.degree. C. for 30 min and settled for 10 min.
The aqueous and organic layers were separated. The aqueous layer
was cooled to 0.degree. C. and acidified to pH.about.2 with 2M HCl.
EtOAc was added and resulting mixture stirred at 25.degree. C. for
30 min. EtOAc layer was separated and the aqueous layer again
extracted with EtOAc. The combined EtOAc layers were washed with DM
water and the organic layer was separated. The organic layer was
washed with sodium chloride solution (1.79 kg of NaCl in 17.94 L of
water). The EtOAc layer was dried over anhydrous Na.sub.2SO.sub.4
and filtered. The EtOAc layer was evaporated below 40.degree. C.
After completion of evaporation, the thick yellow liquid was
subjected to drying by rotary evaporation at 40-45.degree. C. for
1.0 h. Drying was terminated and a pale yellow liquid was obtained
(2.6 kg, yield 95%). .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.85
(d, J=2.4 Hz, 1H), 6.70 (s, 2H), 3.78 (s, 3H), 3.50 (s, 1H).
c) Ethyl 4-((3-chloro-5-methoxyphenyl)thio)-3-oxobutanoate
##STR00104##
[0398] A 100 L reactor was charged with a solution of
3-chloro-5-methoxybenzenethiol (2.6 kg, 1489 mmol) in acetonitrile
(19.5 L). K.sub.2CO.sub.3 (3.09 kg, 2235 mmol) was added to the
reaction mass at 0.degree. C. under N.sub.2 atmosphere and stirred
at same temperature for 10 min (Note: After addition of
K.sub.2CO.sub.3, the reaction mass color changed from pale yellow
to white color). Ethyl 4-chloroacetoacetate (2.7 kg, 1545 mmol) was
added dropwise to the reaction mass at 0-10.degree. C. for 20 min.
The reaction mass was stirred at 25.degree. C. for 1 h under
N.sub.2 atmosphere (Note: After 1 h stirring, the reaction mass
color changed from white to brown color). The reaction was
monitored by TLC (10% EtOAc in pet ether, R.sub.f value of the
product is 0.3). After completion of the reaction, the reaction
mass was cooled to 0.degree. C. DM water was added slowly to the
reaction mass at 0-10.degree. C. EtOAc was added and the resulting
mixture stirred at 25.degree. C. for 10 min. The aqueous and
organic layers were separated. The EtOAc layer was washed with 10%
sodium chloride solution. The aqueous and organic layers were
separated. The EtOAc layer was dried over anhydrous
Na.sub.2SO.sub.4 and filtered. The Na.sub.2SO.sub.4 was washed with
EtOAc. The EtOAc layer was evaporated below 40.degree. C. Drying
was terminated and a dark color liquid was obtained (4.1 kg, yield
91%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 6.90 (d, J=2.4 Hz,
1H), 6.75 (s, 2H), 4.20-4.18 (m, 2H), 3.82 (s, 2H), 3.78 (s, 3H),
3.61 (s, 2H), 1.26 (m. 3H).
d) Ethyl 2-(4-chloro-6-methoxybenzo[b]thiophen-3-yl)acetate and
ethyl 2-(6-chloro-4-methoxybenzo[b]thiophen-3-yl)acetate
##STR00105##
[0400] A 20 L 4-neck round bottom flask was charged with
methanesulfonic acid (9.52 L) and cooled to 0.degree. C. Ethyl
4-(3-chloro-5-methoxyphenylthio)-3-oxobutanoate (4.0 kg, 13211
mmol) was added at 0.degree. C. slowly dropwise into the above
reaction mixture under nitrogen atmosphere for 50 min and stirred
at 0.degree. C. for 20 min (Note: The reaction mixture turns a dark
black color). The reaction mass was slowly allowed to attain
25.degree. C. The reaction mass was stirred at 25.degree. C. for 1
h. Completion of the reaction was monitored by TLC. (10% EtOAc in
pet ether, R.sub.f value of the product is 0.4). After completion
of the reaction, the reaction mass was poured into ice cold water.
EtOAc was added and the resulting mixture stirred at 25.degree. C.
for 10 min. The aqueous and EtOAc layers were separated. The
aqueous layer was again extracted with EtOAc. The combined EtOAc
layers were washed with DM water and the organic layer was
separated. The EtOAc layer was washed with 10% sodium chloride
solution. The EtOAc layer was dried over anhydrous Na.sub.2SO.sub.4
and filtered. The Na.sub.2SO.sub.4 was washed with EtOAc. The EtOAc
was evaporated below 40-45.degree. C. under vacuum. After
completion of evaporation, the thick yellow liquid was subjected to
drying by rotary evaporation at 40-45.degree. C. for 1.0 h. Drying
was terminated and a thick black liquid was obtained (3.5 kg). A
chromatography column was packed with silica gel (12 kg, 100-200
mesh). The crude compound was dissolved in DCM and loaded onto the
column. The mobile phase was run with hexane (50 L) followed by
increasing the polarity from 2-10% EtOAc in hexane (100 L). All
pure fractions (by TLC) collected and concentrated under reduced
pressure at 40-45.degree. C. to afford a mixture of two
regioisomeric compounds (2.0 kg, yield 54% as a mixture, ratio
3:1). LCMS (ES) m/z 285.12 [M+H].sup.+.
e) Ethyl 2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate
##STR00106##
[0402] A 20 L 4-neck round bottom flask was charged with a mixture
of ethyl 2-(4-chloro-6-methoxybenzo[b]thiophen-3-yl)acetate and
ethyl2-(6-chloro-4-methoxybenzo[b]thiophen-3-yl)acetate (mixture
1.0 kg, 3511 mmol) in DCM (9 L). The reaction mass was cooled to
-78.degree. C. BBr.sub.3 (neat, 663.85 mL, 6605.54 mmol) was added
at -78.degree. C. slowly dropwise into the above reaction mixture
under nitrogen atmosphere for 45 min and stirred at -78.degree. C.
for 20 min (Note: The reaction mixture turned brick red and
precipitation was observed on the walls of RB flask). The reaction
mixture was allowed to attain 0.degree. C. and stirred for 2 h
(Note: The reaction mixture turned brick red and precipitation to
wine red color liquid observed). Progress of the reaction was
monitored by TLC. (10% EtOAc in pet ether, R.sub.f value of the
product is 0.3). After completion of the reaction, the reaction
mass was poured into ice cold water slowly dropwise. (Note:
Exothermic reaction while quenching of reaction). DCM was added to
the above reaction mixture and the resulting mixture was stirred at
25.degree. C. for 10 min. The aqueous and DCM layers were
separated. The aqueous layer was again extracted with DCM. The
combined organic layers were washed with DM water and the organic
layer was separated and again washed with DM water. The combined
organic layers were washed with sodium chloride solution. The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered,
and evaporated below 35-40.degree. C. under vacuum. After
completion of evaporation, the orange red color solid was subjected
to drying by rotary evaporation at 40-45.degree. C. for 1.0 h.
Drying was terminated and an orange red color solid was obtained
(800 g, crude) as a mixture of isomers with a ratio of 7:2. The
crude solid (mixture of isomers) was taken up in MTBE (2000 mL) and
stirred at 25.degree. C. for 30 min then cooled to 0.degree. C. for
20 min. The solid was collected by filtration, washed with cold
MTBE (500 mL), and dried by suction to afford ethyl
2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate as an off-white
solid (500 g, yield 52%). .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 9.98 (s, 1H) 7.39 (s, 1H), 7.31 (d, J=2.4 Hz, 1H), 6.87 (d,
J=2.4 Hz, 1H), 4.24 (q, J=9.2 Hz, 2H), 4.08 (s, 2H), 1.30 (t, J=6.9
Hz, 3H). LCMS (ES) m/z 271.15 [M+H].sup.+.
f) Ethyl
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-
benzo[b]thiophen-3-yl)acetate
##STR00107##
[0404] A 5 L 4-neck round bottom flask was charged with a mixture
of (R)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol (107 g, 791.79
mmol, see Scheme-14 for preparation) in THF (2.14 L). Ethyl
2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (214 g, 791.79
mmol) was added at 25.degree. C. (Note: The reaction mixture turns
a brown color). Tri-n-butylphosphine (480 g, 2374.93 mmol) and DIAD
(480 g, 2374.93 mmol) were added at 25.degree. C. The reaction mass
was stirred at 25.degree. C. for 16 h (Note: Reaction mixture turns
a brown color liquid). Completion of the reaction was monitored by
TLC. (50% EtOAc in pet ether, R.sub.f value of the product is 0.4).
After completion of the reaction, the reaction mass was quenched
with ice cold water slowly. EtOAc was added to the above reaction
mixture and the resulting mixture stirred at 25.degree. C. for 10
min. The Aqueous and EtOAc layers were separated. The aqueous layer
was again extracted with EtOAc. The aqueous and EtOAc layers were
separated. The combined organic layers were washed with DM water
and the organic layer was separated. The organic layer was washed
with sodium chloride solution. The EtOAc layer was dried over
anhydrous Na.sub.2SO.sub.4 and filtered. EtOAc was evaporated below
40-45.degree. C. under vacuum. After completion of evaporation, the
black color liquid was subjected to drying by rotary evaporation at
40-45.degree. C. for 1.0 h. Drying was terminated and a brown color
gum was obtained (400 g). A chromatography column was packed with
silica gel (700 g, 100-200 mesh). The crude compound was dissolved
in DCM and adsorbed onto silica gel (300 g) and loaded onto the
column. The mobile phase was run with n-hexane (20 L) followed by
increasing the polarity from 2-10% EtOAc in hexane (60 L). All pure
fractions (by TLC) were collected and concentrated under reduced
pressure at 40-45.degree. C. to obtain pure product (210 g, yield
68%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.60 (d, J=4.8 Hz,
1H), 7.70 (d, J=4.8 Hz, 1H), 7.49 (s, 1H), 7.23-7.28 (m, 2H), 7.13
(d, J=2.1 Hz, 1H), 5.81-5.80 (m, 1H), 4.21-4.20 (q, 2H), 4.01 (s,
2H), 3.30-3.20 (m, 1H), 3.10-3.00 (m, 1H), 2.66-2.68 (m, 1H),
2.12-2.10 (m, 1H), 1.30 (t, J=6.9 Hz, 3H). LCMS (ES) m/z 388.08
[M+H].sup.+.
g)
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[-
b]thiophen-3-yl)acetic Acid
##STR00108##
[0406] A 10 L 4-neck round bottom flask was charged with (S)-ethyl
2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]thio-
phen-3-yl)acetate (400 g, 1031 mmol) in THF (2 L), methanol (2 L),
and DM water (1 L) at 25.degree. C. Lithium hydroxide (123 g, 5156
mmol) was added at 25.degree. C. slowly into the above reaction
mixture (Note: The reaction mixture turned a brown color). The
reaction mass was stirred at 25.degree. C. for 2 h. The reaction
was monitored by TLC (80% EtOAc in pet ether, R.sub.f value of the
product is 0.2). After completion of the reaction, the reaction
mass was poured into ice cold water. The reaction mass was
acidified with 10% NaHSO.sub.4 pH.about.6 solution and a white
precipitate formed. The solid was collected by filtration, washed
with DM water, and dried by suction. The solid was washed with
diethyl ether and dried for 2 h. The solid was further dried by
rotary evaporation below 45-50.degree. C. under vacuum for 10 h.
The solid compound was further triturated with MTBE, filtered, and
dried to obtain the desired compound as an off-white solid (190 g,
yield 51%). .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 12.3-12.4
(br, 1H), 8.50-8.52 (dd, J=1.2, 4.8 Hz, 1H), 7.78-7.83 (m, 2H),
7.49 (s, 1H), 7.23-7.28 (m, 1H), 7.13 (d, J=2.1 Hz, 1H), 6.00-6.04
(m, 1H), 4.01 (s, 2H), 3.06-3.11 (m, 1H), 2.95-2.98 (m, 1H),
2.66-2.68 (m, 1H), 2.09-2.12 (m, 1H). LCMS (ES) m/z 359.98
[M+H].sup.+. Chiral HPLC: 99.70%.
##STR00109##
a) 6,7-Dihydro-5H-cyclopenta[b]pyridin-5-one
##STR00110##
[0408] A 50 L glass reactor was charged KMnO.sub.4 (1857 g, 11748.0
mmol). Water (35 L) was charged to the reaction mass. The reaction
mass was stirred for 30 min (Note: KMnO.sub.4 completely soluble in
water). Another 100 L reactor was charged
6,7-dihydro-5H-cyclopenta[b]pyridine (700 g, 5874 mmol) in
tert-butanol (17.5 L) at 25.degree. C. MgSO.sub.4 (1414 g, 11748
mmol) was charged to the above reaction mixture. The reaction
mixture was cooled to 20-25.degree. C. with ice water. KMnO.sub.4
solution was added dropwise for 2 h (Note: Slight exothermic was
observed and temperature maintained below 30.degree. C. with ice
water). The reaction mixture was maintained at 30.degree. C. for 3
h. Progress of the reaction was monitored by TLC/LCMS (50% EtOAc in
pet ether, R.sub.f value of the product is 0.3). After completion
of the reaction, EtOAc was added to the above reaction mixture and
the resulting mixture stirred at 25.degree. C. for 10 min. The
aqueous and EtOAc layers were separated. The aqueous layer was
again extracted with EtOAc. The combined organic layers were washed
with DM water and the organic layer was separated. The combined
organic layers were washed with sodium chloride solution. The EtOAc
layer was dried over anhydrous Na.sub.2SO.sub.4 and filtered. EtOAc
was evaporated below 40-45.degree. C. under vacuum. After
completion of evaporation, the brown color liquid was subjected to
drying by rotary evaporation at 40-45.degree. C. for 1.0 h. Drying
was terminated and a brown color gum was obtained (600 g). A
chromatography column was packed with silica gel (4.0 kg, 100-200
mesh). The crude compound was dissolved in DCM and adsorbed onto
silica gel (1.0 kg) and loaded onto the column. The mobile phase
was run with n-hexane (25 L) followed by increasing the polarity
from 2-10% EtOAc in hexane (100 L). All pure fractions (TLC) were
collected and concentrated under reduced pressure at 40-45.degree.
C. to give 6,7-dihydro-5H-cyclopenta[b]pyridin-5-one as a brown
thick gum (270 g, yield 34%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.45 (dd, J=1.5, 4.5 Hz, 1H), 8.02 (dd, J=2.0, 8.0 Hz, 1H),
7.46-7.44 (m, 1H), 3.19-3.16 (m, 2H), 2.73-2.71 (m, 2H). LCMS (ES)
m/z 134.07 [M+H].sup.+.
b) (R)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol
##STR00111##
[0410] A 5 L 4-neck round bottom flask was charged
6,7-dihydro-5H-cyclopenta[b]pyridine-5-one (100 g, 751.04 mmol) in
EtOAc (2 L). TEA (523 mL, 3755.2 mmol) was added at 25.degree. C.
slowly into the above reaction mixture. The reaction mass was
cooled to 0.degree. C. and formic acid (346 g, 7510.4 mmol) was
added dropwise over 30 min (Note: thick white fumes were observed).
The above reaction mixture was stirred at 0.degree. C. for 30 min.
RuCl[(R,R)-Tsdpen](mesitylene) (9.36 g, 15.02 mmol) was added at
0.degree. C. The reaction mixture was maintained at 45.degree. C.
for 16 h. Progress of the reaction was monitored by TLC. (50% EtOAc
in pet ether, R.sub.f value of the product is 0.4). After
completion of the reaction, the reaction mass was directly
evaporated by rotary evaporation below 40-45.degree. C. under
vacuum. A chromatography column was packed with silica gel (500 g,
100-200 mesh). The crude compound was directly loaded onto the
column. The mobile phase was run with n-hexane (25 L) followed by
increasing the polarity from 2-80% EtOAc in hexane (50 L). All pure
fractions (by TLC) were collected and concentrated under reduced
pressure at 40-45.degree. C. to give a gummy liquid, which was
triturated with diethyl ether (2.times.100 mL) and filtered by
suction to afford (R)-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ol as a
pale brown solid (80 g, yield 79%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.45 (dd, J=1.2, 3.6 Hz, 1H), 7.71 (d, J=7.6
Hz, 1H), 7.14 (q, J=4.8 Hz, 1H), 5.29 (d, J=5.6 Hz, 1H), 3.20-3.12
(m, 2H), 2.98-2.89 (m, 1H), 2.62-2.54 (m, 1H), 2.03-1.98 (m, 1H).
LCMS (ES) m/z 136.17 [M+H].sup.+.
Crystalline Compound of Example 1a,
(S)-2-(4-chloro-6-((6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benzo[b]-
thiophen-3-yl)acetic Acid
[0411] The X-ray powder diffraction (XRPD) pattern of this material
is shown in FIG. 1 and a summary of the diffraction angles and
d-spacings is given in Table I below. The XRPD analysis was
conducted on a PANanalytical X'Pert Pro Diffractometer on Si
zero-background wafers using X'celerator.TM. RTMS (Real Time
Multi-Strip) detector. The acquisition conditions included: Cu
K.sub..alpha. radiation, generator tension: 45 kV, generator
current: 40 mA, step size: 0.0167.degree. 2.theta.. Configuration
on the incidental beam side: 10 mm programmable divergence slit,
0.02 rad Soller slits, anti-scatter slit (0.5.degree.), and 10 mm
beam mask. Configuration on the diffracted beam side: 10 mm
programmable anti-scatter slit assembly (X'celerator module) and
0.02 rad Soller slit.
TABLE-US-00004 TABLE I Diff. Angle [.degree.2.theta.] d-spacing
[.ANG.] 5.876 15.0412 13.6014 6.51043 14.0485 6.30422 14.3468
6.17377 21.8816 4.06196 22.461 3.95847 23.0524 3.85824 23.3301
3.81294 24.1261 3.6889 24.5102 3.63196 24.6985 3.6047 25.6652
3.47107 26.0846 3.41621 26.6086 3.35011 27.4121 3.25371
[0412] The differential scanning calorimetry (DSC) thermogram of
this material was recorded on a TA Instruments Discovery
Differential Scanning Calorimeter equipped with an autosampler and
a refrigerated cooling system under 40 mL/min N.sub.2 purge and is
shown in FIG. 2. The experiments were conducted using a heating
rate of 10.degree. C./min to final temperature of 350.degree. C. in
a lightly crimped aluminum pan.
[0413] The thermogravimetric analysis (TGA) thermogram of this
material was recorded on a TA Instruments Discovery
Thermogravimetric Analyzer and is shown in FIG. 2. The experiments
were conducted under N.sub.2 purge and a heating rate of 10.degree.
C./min to final temperature of 350.degree. C. in an open aluminum
pan.
[0414] This compound has a simple single melting event in DSC, with
onset temperature of 220.8 C, peak temperature of 223.4 C and
melting enthalpy of 120 J/g. The determination of melting enthalpy
is not reliable due to the immediate thermal decomposition post
melting. The compound exhibited negligible weight loss by loss by
TGA prior to the decomposition event. A person skilled in the art
would recognize that the onset temperature, peak temperature, and
enthalpy of the endotherm may vary depending on the experimental
conditions.
Example 2
##STR00112##
[0415] Preparation of
2-(4-chloro-6-((2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)ben-
zo[b]thiophen-3-yl)acetic Acid
a) Ethyl
2-(4-chloro-6-(2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-ylo-
xy)benzo[b]thiophen-3-yl)acetate
##STR00113##
[0417] The crude
5-Chloro-2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridine (25 g) was
dissolved in DMF (250 mL) at RT and to this ethyl
2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (25 g, 92 mmol)
and K.sub.2CO.sub.3 (63.8 g, 462 mmol) were added at RT. The
reaction mixture was heated to 80.degree. C. for 2 h. The mixture
was diluted with water and extracted with EtOAc. The organic layer
was washed successively with water and brine, dried over MgSO.sub.4
and concentrated in vacuo to get crude. The crude residue was
purified by silica gel column chromatography (EtOAc/hexane) to give
the title compound (25 g) as an off white solid. LCMS (ES) m/z
402.17 [M+H].sup.+.
b)
2-(4-Chloro-6-((2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)b-
enzo[b]thiophen-3-yl)acetic Acid
##STR00114##
[0419] The title compound was obtained in a same manner as the
procedure in Example 1, Step b by using ethyl
2-(4-chloro-6-(2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yloxy)benzo-
[b]thiophen-3-yl)acetate as an off white solid. .sup.1H NMR (300
MHz, DMSO-d.sub.6): .delta. 7.76 (d, J=2.1 Hz, 1H), 7.68 (d, J=8.1
Hz, 1H), 7.47 (s, 1H), 7.11-7.08 (m, 2H), 5.98-5.95 (m, 1H), 4.00
(s, 2H), 3.17-3.01 (m, 1H), 2.93-2.83 (m, 1H), 2.67-2.59 (m, 1H),
2.47 (s, 3H), 2.13-2.04 (m, 1H); LCMS (ES) m/z 374.09 [M+H].sup.+.
Chiral HPLC: 49.85%:50.14%.
Analytical SFC condition
Column/dimensions: Chiralpak AD-H (250.times.4.6) mm, 5.mu.
% CO.sub.2:60.0%
[0420] % Co solvent: 40.0% (100% Methanol) Total Flow: 4.0
g/min
Back Pressure: 100 bar
Temperature: 30.0.degree. C.
UV: 235 nm
Preparative SFC Condition
Column/dimensions: Lux Amylose-1 (250.times.30) mm, 5.mu.
% CO.sub.2: 55.0%
[0421] % Co solvent: 45.0% (100% Methanol) Total Flow: 90.0
g/min
Back Pressure: 100.0 bar
UV: 235 nm
[0422] Stack time: 5.3 min
Load/Inj: 82.0 mg
[0423] Retention time: Peak 1-3.02 min, Peak 2-4.93 min.
Purity: Peak 1-99.91%, Peak 2-99.24%.
[0424] Solubility: Methanol (660 mL)+12 ml DEA Instrument details:
Make/Model: Thar SFC-200-002
[0425] Example 2a:
(S)-2-(4-chloro-6-((2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetic acid (5.1 g, 30.7% yield). .sup.1H
NMR (300 MHz, DMSO-d.sub.6): .delta. 7.76 (d, J=2.4 Hz, 1H), 7.68
(d, J=7.8 Hz, 1H), 7.47 (s, 1H), 7.11-7.08 (m, 2H), 5.98-5.95 (m,
1H), 4.00 (s, 2H), 3.06-2.89 (m, 2H), 2.64-2.62 (m, 1H), 2.47 (s,
3H), 2.10-2.06 (m, 1H). LCMS (ES) m/z 374.09 [M+H].sup.+. Chiral
HPLC: 99.91%.
[0426] Example 2b:
(R)-2-(4-chloro-6-((2-methyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetic acid (3.0 g, 18.24% yield). .sup.1H
NMR (300 MHz, DMSO-d.sub.6): .delta. 7.76 (d, J=2.4 Hz, 1H), 7.68
(d, J=7.8 Hz, 1H), 7.47 (s, 1H), 7.11-7.08 (m, 2H), 5.98-5.95 (m,
1H), 4.00 (s, 2H), 3.06-2.89 (m, 2H), 2.64-2.62 (m, 1H), 2.47 (s,
3H), 2.10-2.06 (m, 1H). LCMS (ES) m/z 374.24 [M+H].sup.+. Chiral
HPLC: 99.24%.
Example 3
##STR00115##
[0427] Preparation of
2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetic Acid
a) Ethyl
2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin--
5-yl)oxy)benzo[b]thiophen-3-yl)acetate
##STR00116##
[0429] To the crude
2,5-dichloro-6,7-dihydro-5H-cyclopenta[b]pyridine (100 mg)
dissolved in DMF was added to a stirred solution of ethyl
2-(4,7-dichloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (100 mg,
0.328 mmol) and K.sub.2CO.sub.3 (181 mg, 1.311 mmol) in DMF (5 mL)
at ambient temperature and then heated to 100 for 2 h. After TLC
analysis the reaction mixture was diluted with water and extracted
with EtOAc. The organic layer was washed with water, brine, dried
over Na.sub.2SO.sub.4 and concentrated under reduced pressure to
afford crude. The crude was purified by silica gel chromatography
using 30% EtOAc/pet ether as an eluent to afford ethyl
2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetate (60 mg, 40.0% yield) as an oily
liquid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.66 (d, J=8.0
Hz, 1H), 7.31-7.18 (m, 3H), 5.78-5.75 (m, 1H), 4.22 (q, J=8 Hz,
2H), 4.09 (s, 2H), 3.35-3.27 (m, 1H), 3.08-.3.00 (m, 1H), 2.69-2.64
(m, 1H), 2.42-2.39 (m, 1H), 1.29 (t, J=8 Hz, 3H). LCMS (ES) m/z
456.79 [M+H].sup.+.
b)
2-(4,7-Dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)o-
xy)benzo[b]thiophen-3-yl)acetic Acid
##STR00117##
[0431] To a stirred solution of ethyl
2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetate (60 mg, 0.131 mmol) in methanol (2
mL), THF (2 mL) and water (2 mL), LiOH (6.29 mg, 0.263 mmol) was
added at ambient temperature and stirred for 4 h. After TLC
analysis the reaction mixture was evaporated to remove solvents and
the crude was cooled to 0.degree. C., acidified with saturated
citric acid solution (p.sup.H.about.5). Obtained solids were
filtered and dried well to afford
2-(4,7-dichloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy-
)benzo[b]thiophen-3-yl)acetic acid (46 mg, 81% yield) as an
off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.47
(brs, 1H), 7.83 (d, J=8.0 Hz, 1H), 7.65-7.61 (m, 2H), 7.39 (d,
J=8.0 Hz, 1H) 6.10-6.08 (m, 1H), 4.03 (s, 2H), 3.19-3.11 (m, 1H),
2.99-2.92 (m, 1H), 2.71-2.62 (m, 1H), 2.20-2.10 (m, 1H). LCMS (ES)
m/z 428 [M+H].sup.+. Chiral HPLC: 48.83%:51.16%.
Analytical SFC Conditions
[0432] Column/dimensions: Chiralpak AD-H (4.6.times.250 mm),
5.mu.
% CO.sub.2: 60.0%
[0433] % Co solvent: 40.0% (100% MeOH) Total Flow: 4.0 g/min
Back Pressure: 100 bar
Temperature: 30.degree. C.
UV: 235 nm
Preparative SFC Conditions
[0434] Column/dimensions: Chiralpak AD-H (30.times.250 mm),
5.mu.
% CO.sub.2: 50.0%
[0435] % Co solvent: 50.0% (100% MeOH) Total Flow: 90.0 g/min
Back Pressure: 100.0 bar
UV: 235 nm
[0436] Stack time: 15.3 min
Load/Inj: 48.3 mg
[0437] Retention time: Peak 1-3.59 min, Peak 2-13.34 min.
Purity: Peak 1-99.06%, Peak 2-99.73%.
[0438] Solubility: 20 ml MeOH+few drops of methanolic ammonia
solution Instrument details: Make/Model: SFC-200-003
Chiral Separation of Example 3
Example 3a (First Eluted Enantiomer):
[0439] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.47 (brs, 1H),
7.83 (d, J=8.0 Hz, 1H), 7.65-7.61 (m, 2H), 7.39 (d, J=8.0 Hz, 1H)
6.10-6.08 (m, 1H), 4.03 (s, 2H), 3.19-3.11 (m, 1H), 2.99-2.92 (m,
1H), 2.71-2.62 (m, 1H), 2.20-2.10 (m, 1H). LCMS (ES) m/z 427.9
[M+H].sup.+. Chiral HPLC: 99.06%.
Example 3b (Second Eluted Enantiomer):
[0440] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.47 (brs, 1H),
7.83 (d, J=8.0 Hz, 1H), 7.65-7.61 (m, 2H), 7.39 (d, J=8.0 Hz, 1H)
6.10-6.08 (m, 1H), 4.03 (s, 2H), 3.19-3.11 (m, 1H), 2.99-2.92 (m,
1H), 2.71-2.62 (m, 1H), 2.20-2.10 (m, 1H). LCMS (ES) m/z 427.99
[M+H].sup.+. Chiral HPLC: 99.77%
Example 4
##STR00118##
[0441] Preparation of
2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyrid-
in-5-yl)oxy)benzo[b]thiophen-3-yl)acetic Acid
a) Ethyl
2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta-
[b]pyridin-5-yl)oxy)benzo[b]thiophen-3-yl)acetate
##STR00119##
[0443] To the crude
5-Chloro-2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridine
(120 mg) dissolved in DMF was added to a stirred solution of ethyl
2-(4,7-dichloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (100 mg,
0.328 mmol) and K.sub.2CO.sub.3 (181 mg, 1.311 mmol) in DMF (5 mL)
at ambient temperature and then heated to 100.degree. C. for 2 h.
After TLC analysis the reaction mixture was diluted with water and
extracted with EtOAc. The organic layer was washed with water,
brine, dried over Na.sub.2SO.sub.4 and concentrated under reduced
pressure to afford crude. The crude was purified by silica gel
chromatography using 30% EtOAc/pet. ether as an eluent to afford
ethyl
2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyrid-
in-5-yl)oxy)benzo[b]thiophen-3-yl)acetate (50 mg, 30.9% yield) as
an oily liquid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.88 (d,
J=8.0 Hz, 1H), 7.58 (d, J=8 Hz, 1H), 7.26 (s, 1H), 7.20 (s, 1H),
5.82-5.78 (m, 1H), 4.22 (q, J=8 Hz, 2H), 4.10 (s, 2H), 3.42-3.35
(m, 1H), 3.18-.3.10 (m, 1H), 2.75-2.69 (m, 1H), 2.46-2.41 (m, 1H),
1.27 (t, J=8 Hz, 3H). LCMS (ES) m/z 490.67 (M+H).sup.+.
b)
2-(4,7-Dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyr-
idin-5-yl)oxy)benzo[b]thiophen-3-yl)acetic acid
##STR00120##
[0445] To a stirred solution of ethyl
2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyrid-
in-5-yl)oxy)benzo[b]thiophen-3-yl)acetate (50 mg, 0.102 mmol) in
methanol (2 mL), THF (2 mL) and water (2 mL), LiOH (4.88 mg, 0.204
mmol) was added at ambient temperature and stirred for 4 h. After
TLC analysis the reaction mixture was evaporated to remove solvents
and the crude was cooled to 0.degree. C., acidified with saturated
citric acid solution (pH.about.5). Obtained solids were filtered
and dried well to afford
2-(4,7-dichloro-6-((2-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyrid-
in-5-yl)oxy)benzo[b]thiophen-3-yl)acetic acid (40 mg, 85% yield) as
an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.
12.48 (brs, 1H), 8.08 (d, J=7.5 Hz, 1H), 7.80 (d, J=7.8 Hz, 1H),
7.70 (s, 1H), 7.63 (s, 1H), 6.21-6.17 (m, 1H), 4.04 (s, 2H),
3.31.3.11 (m, 1H), 3.08-3.01 (m, 1H), 2.79-2.67 (m, 1H), 2.28-2.18
(m, 1H). LCMS (ES) m/z 462.17 [M+H].sup.+. Chiral HPLC 47.52%:
52.47%.
Analytical SFC Condition
[0446] Column/dimensions: Chiralpak AD-H (4.6.times.250 mm),
5.mu.
% CO.sub.2:60.0%
[0447] % Co solvent: 40.0% (100% methanol) Total Flow: 4.0
g/min
Back Pressure: 100 bar
Temperature: 30.degree. C.
UV: 234 nm
Preparative SFC Condition
[0448] Column/dimensions: Chiralpak AD-H (30.times.250 mm),
5.mu.
% CO.sub.2: 60.0%
[0449] % Co solvent: 40.0% (100% methanol) Total Flow: 90.0
g/min
Back Pressure: 100.0 bar
UV: 234 nm
[0450] Stack time: 8.5 min
Load/Inj: 50.0 mg
[0451] Retention time: Peak 1-1.82 min, Peak 2-5.75 min.
Purity: Peak 1-99.68%, Peak 2-99.82%.
Solubility: Methanol+ACN
[0452] Instrument details: Make/Model: SFC-PIC SOLUTION
Chiral Separation of Example 4
Example 4a (First Eluted Enantiomer):
[0453] .sup.1H NMR (500 MHz, CDCl.sub.3): .delta. 7.88 (d, J=8.0
Hz, 1H), 7.58 (d, J=7.5 Hz, 1H), 7.30 (s, 1H), 7.22 (s, 1H),
5.84-5.82 (m, 1H), 4.17 (s, 2H), 3.41-3.36 (m, 1H), 3.18-3.11 (m,
1H), 2.75-2.69 (m, 1H), 2.46-2.40 (m, 1H). LCMS (ES) m/z 462.14
[M+H].sup.+, HPLC: 99.69%. Chiral HPLC: 99.81%.
Example 4b (Second Eluted Enantiomer):
[0454] .sup.1H NMR (500 MHz, CDCl.sub.3): .delta. 7.88 (d, J=8.0
Hz, 1H), 7.58 (d, J=7.5 Hz, 1H), 7.30 (s, 1H), 7.22 (s, 1H),
5.83-5.81 (m, 1H), 4.15 (s, 2H), 3.40-3.35 (m, 1H), 3.17-3.11 (m,
1H), 2.73-2.70 (m, 1H), 2.44-2.39 (m, 1H). LCMS (ES) m/z 462.11
[M+H].sup.+, HPLC: 99.94%, Chiral HPLC: 99.82%
Example 5
##STR00121##
[0455] Preparation of
2-(4-chloro-7-fluoro-6-((6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]pyridi-
n-3-yl)oxy)benzo[b]thiophen-3-yl)acetic Acid
a) Ethyl
2-(4-chloro-7-fluoro-6-((6-(trifluoromethyl)-2,3-dihydrofuro[2,3--
b]pyridin-3-yl)oxy)benzo[b]thiophen-3-yl)acetate
##STR00122##
[0457] To a stirred solution of ethyl
2-(4-chloro-7-fluoro-6-hydroxybenzo[b]thiophen-3-yl)acetate (0.141
g, 0.487 mmol),
6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]pyridin-3-ol (0.100 g,
0.487 mmol) and ADDP (0.123 g, 0.487 mmol) in THF (10 mL) was added
tri-n-butylphosphine (0.120 mL, 0.487 mmol) at RT. The reaction
mixture was stirred at room temperature for 48 h, filtered through
Celite.RTM. and evaporated under reduced pressure to afford crude
product as yellow liquid, which was purified by flash column
chromatography on 100-200 silica gel, using 30% EtOAc-Pet ether as
an eluent to obtained ethyl
2-(4-chloro-7-fluoro-6-((6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]-
pyridin-3-yl)oxy)benzo[b]thiophen-3-yl)acetate (0.090 g, 37.5%
yield) as pale yellow liquid. LCMS (ES) m/z 476.05 [M+H].sup.+.
b)
2-(4-Chloro-7-fluoro-6-((6-(trifluoromethyl)-2,3-dihydrofuro[2,3-b]pyri-
din-3-yl)oxy)benzo[b]thiophen-3-yl)acetic acid
##STR00123##
[0459] The title compound was prepared as a white solid according
to the procedures of examples XX as white solid. .sup.1H NMR (400
MHz, DMSO-d.sub.6): .delta. 8.09 (d, J=7.6 Hz, 1H), 7.60 (d, J=6.8
Hz, 1H), 7.51 (d, J=8.0 Hz, 1H), 7.46 (s, 1H), 6.31-6.30 (m, 1H),
4.94-4.83 (m, 2H), 3.77 (s, 2H). LCMS (ES) m/z 448.16 [M+H].sup.+.
Chiral HPLC: 50.55%:49.45%.
Analytical SFC Condition
[0460] Column/dimensions: Chiralcel OJ-H (4.6.times.250 mm),
5.mu.
% CO.sub.2: 80.0%
[0461] % Co solvent: 20.0% (100% MeOH) Total Flow: 4.0 g/min
Back Pressure: 100 bar
Temperature: 30.degree. C.
UV: 214 nm
Preparative SFC Condition
[0462] Column/dimensions: Chiralcel OJ-H (21.times.250 mm),
5.mu.
% CO.sub.2: 90.0%
[0463] % Co solvent: 10.0% (100% MeOH) Total Flow: 60.0 g/min
Back Pressure: 100.0 bar
UV: 214 nm
[0464] Stack time: 4.3 min
Load/Inj: 2.5 mg
[0465] Retention time: Peak 1-2.93 min, Peak 2-4.89 min.
Purity: Peak 1-99.59%, Peak 2-99.30%.
Solubility: MeOH
[0466] Instrument details: Make/Model: SFC-80
Chiral Separation of Example 5
Example 5a (First Eluted Enantiomer):
[0467] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.09 (d, J=7.6
Hz, 1H), 7.60 (d, J=6.8 Hz, 1H), 7.51 (d, J=8.0 Hz, 1H), 7.46 (s,
1H), 6.31-6.30 (m, 1H), 4.94-4.83 (m, 2H), 3.77 (s, 2H). LCMS (ES)
m/z 447.82 [M+H].sup.+. Chiral HPLC: 99.59%.
Example 5b (Second Eluted Enantiomer):
[0468] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.09 (d, J=7.6
Hz, 1H), 7.60 (d, J=6.8 Hz, 1H), 7.51 (d, J=8.0 Hz, 1H), 7.46 (s,
1H), 6.31-6.30 (m, 1H), 4.94-4.83 (m, 2H), 3.77 (s, 2H). LCMS (ES)
m/z 448.26 [M+H].sup.+. Chiral HPLC: 99.30%.
Example 6
##STR00124##
[0469] Preparation of
2-(4-chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thiophen-3-
-yl)acetic Acid
a) Ethyl
2-(4-chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]th-
iophen-3-yl)acetate
##STR00125##
[0471] The title compound was obtained in a same manner as the
procedure in Example 5, Step a by using
2,3-dihydrofuro[2,3-b]pyridin-3-ol and
5-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine, LCMS (ES) m/z 390.34
(M+H).sup.+.
b)
2-(4-Chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thiophen-
-3-yl)acetic acid
##STR00126##
[0473] To a solution of ethyl
2-(4-chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thiophen-3-
-yl)acetate (1.7 g, 4.36 mmol) in Methanol (16 mL), THF (8 mL) and
Water (16 mL) was added LiOH (0.522 g, 21.80 mmol) at rt and
stirred for 2 h at same temperature. Reaction mixture was acidified
with citric acid solution (nearly pH=6-7), filtered the solid
precipitated and dried under vacuum to get
2-(4-chloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thiophen-3-
-yl)acetic acid (1 g, 2.73 mmol, 62.7% yield) as an white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 12.39 (brs, 1H), 8.19
(dd, J=2.0, 5.2 Hz, 1H), 7.92 (dd, J=1.6, 7.2 Hz, 1H), 7.79 (d,
J=2.4 Hz, 1H), 7.51 (s, 1H), 7.15 (d, J=2.8 Hz, 1H), 7.00 (dd,
J=4.8, 7.4 Hz, 1H), 6.23-6.21 (m, 1H), 4.85-4.81 (m, 1H), 4.62-4.59
(m, 1H), 4.01 (s, 2H). ESI-MS m/z 362.13 [M+H].sup.+. Chiral HPLC:
48.09%: 50.68%.
Analytical SFC Condition
[0474] Column/dimensions: Chiralpak AS-H (4.6.times.250 mm),
5.mu.
% CO.sub.2: 65.0%
[0475] % Co solvent: 35.0% (100% methanol) Total Flow: 3.0
g/min
Back Pressure: 100 bar
Temperature: 30.degree. C.
UV: 235 nm
Preparative SFC Condition
[0476] Column/dimensions: Chiralpak AS-H (30.times.250 mm),
5.mu.
% CO.sub.2: 65.0%
[0477] % Co solvent: 35.0% (100% methanol) Total Flow: 100.0
g/min
Back Pressure: 100.0 bar
UV: 235 nm
[0478] Stack time: 6.5 min
Load/Inj: 18.0 mg
[0479] Retention time: Peak 1-3.41 min, Peak 2-4.92 min.
Purity: Peak 1-99.82%, Peak 2-99.50%.
Solubility: Methanol+ACN
[0480] Instrument details: Make/Model: SFC-200-004
(PIC-Solution)
Chiral Separation of Example 6
Example 6a (First Eluted Enantiomer):
[0481] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 12.39 (brs,
1H), 8.19 (dd, J=2.0, 5.2 Hz, 1H), 7.92 (dd, J=1.6, 7.2 Hz, 1H),
7.79 (d, J=2.4 Hz, 1H), 7.51 (s, 1H), 7.15 (d, J=2.8 Hz, 1H), 7.00
(dd, J=4.8, 7.4 Hz, 1H), 6.23-6.21 (m, 1H), 4.87-4.80 (m, 1H),
4.64-4.57 (m, 1H), 4.01 (s, 2H). LCMS (ES) m/z 362.13 [M+H].sup.+.
Chiral HPLC purity: 99.80%.
Example 6b (Second Eluted Enantiomer):
[0482] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 12.39 (brs,
1H), 8.19 (dd, J=2.0, 5.2 Hz, 1H), 7.92 (dd, J=1.6, 7.2 Hz, 1H),
7.79 (d, J=2.4 Hz, 1H), 7.51 (s, 1H), 7.15 (d, J=2.8 Hz, 1H), 7.00
(dd, J=4.8, 7.4 Hz, 1H), 6.23-6.21 (m, 1H), 4.87-4.80 (m, 1H),
4.64-4.57 (m, 1H), 4.01 (s, 2H). LCMS (ES) m/z 362.13 [M+H].sup.+.
Chiral HPLC purity: 99.50%.
Example 7
##STR00127##
[0483] Preparation of
2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thioph-
en-3-yl)acetic Acid
a) Ethyl
2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[-
b]thiophen-3-yl)acetate
##STR00128##
[0485] To a stirred solution of ADDP (827 mg, 3.28 mmol) in dry THF
(2 mL), tri-n-butylphosphine (1.079 mL, 4.38 mmol) was slowly added
at ambient temperature. After decolorisation was observed,
2,3-dihydrofuro[2,3-b]pyridin-3-ol (300 mg, 2.188 mmol) was added
and stirred for 5 min. Finally ethyl
2-(4,7-dichloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (668 mg,
2.188 mmol) was added. The reaction mass was stirred at the same
temperature for 24 h. After TLC analysis the reaction mixture was
diluted with water and extracted with EtOAc. The organic layer was
washed with water, brine, dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to afford crude. The crude was
purified by silica gel chromatography using 30% EtOAc/pet ether as
an eluent to afford ethyl
2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thioph-
en-3-yl)acetate (120 mg) as a colorless gummy liquid. LCMS (ES) m/z
424.14 [M+H].sup.+
b)
2-(4,7-Dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thio-
phen-3-yl)acetic acid
##STR00129##
[0487] To a stirred solution of ethyl
2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thioph-
en-3-yl)acetate (120 mg, 0.283 mmol) in methanol (1 mL), THF (1 mL)
and water (1.000 mL), lithium hydroxide (20.32 mg, 0.848 mmol) was
added at ambient temperature and stirred for 4 h. After TLC
analysis the reaction mixture was evaporated to remove solvents and
the crude was cooled to 0.degree. C. and acidified with saturated
citric acid solution (p.sup.H.about.5). Obtained solids were
filtered and dried well to afford
2-(4,7-dichloro-6-((2,3-dihydrofuro[2,3-b]pyridin-3-yl)oxy)benzo[b]thioph-
en-3-yl)acetic acid (72 mg, 63.7% yield) as an off-white solid.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.21-8.20 (m, 1H),
7.85-7.84 (m, 1H), 7.63 (s, 1H), 7.54 (s, 1H), 7.02-6.99 (m, 1H),
6.31 (d, J=4.5 Hz, 1H), 4.83-4.74 (m, 1H), 4.67-4.64 (m, 1H), 3.92
(s, 2H). LCMS (ES) m/z 396.25 [M+H].sup.+. Chiral HPLC: 48.99%:
49.56%.
Analytical SFC Condition
[0488] Column/dimensions: Chiralpak AD-H (4.6.times.250 mm),
5.mu.
% CO.sub.2:60.0%
[0489] % Co solvent: 40.0% (100% MeOH) Total Flow: 4.0 g/min
Back Pressure: 100 bar
Temperature: 30.degree. C.
UV: 214 nm
Preparative SFC Condition
[0490] Column/dimensions: Chiralpak AD-H (30.times.250 mm),
5.mu.
% CO.sub.2: 60.0%
[0491] % Co solvent: 40.0% (100% MeOH) Total Flow: 90.0 g/min
Back Pressure: 90.0 bar
UV: 214 nm
[0492] Stack time: 8.0 min
Load/Inj: 25.0 mg
[0493] Retention time: Peak 1-2.73 min, Peak 2-5.34 min.
Purity: Peak 1-99.81%, Peak 2-98.63%
Solubility: MeOH+Acetonitrile
Chiral Separation of Example 7
Example 7a (First Eluted Enantiomer):
[0494] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.89 (brs, 1H),
8.21-8.20 (m, 1H), 7.85-7.84 (m, 1H), 7.63 (s, 1H), 7.54 (s, 1H),
7.02-6.99 (m, 1H), 6.31 (d, J=4.5 Hz, 1H), 4.83-4.74 (m, 1H),
4.67-4.64 (m, 1H), 3.92 (s, 2H). LCMS (ES) m/z 396.15 [M+H].sup.+.
Chiral purity: 99.81%.
Example 7b (Second Eluted Enantiomer):
[0495] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.89 (brs, 1H),
8.21-8.20 (m, 1H), 7.85-7.84 (m, 1H), 7.63 (s, 1H), 7.54 (s, 1H),
7.02-6.99 (m, 1H), 6.31 (d, J=4.5 Hz, 1H), 4.83-4.74 (m, 1H),
4.67-4.64 (m, 1H), 3.92 (s, 2H). LCMS (ES) m/z 396.25 [M+H].sup.+.
Chiral purity: 98.63%
Examples 8 and 9
##STR00130##
[0496] Preparation of
2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benz-
o[b]thiophen-3-yl)acetic acid (Example 8)
a) Ethyl
2-(4-chloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl-
)oxy)benzo[b]thiophen-3-yl)acetate
##STR00131##
[0498] 2,5-Dichloro-6,7-dihydro-5H-cyclopenta[b]pyridine (556 mg,
2.95 mmol) was added to the stirred solution of K.sub.2CO.sub.3
(1225 mg, 8.86 mmol) and ethyl
2-(4-chloro-6-hydroxybenzo[b]thiophen-3-yl)acetate (800 mg, 2.95
mmol) in DMF (20 mL) at 0.degree. C. and the mixture was stirred at
60.degree. C. for 2 h. The reaction mixture was diluted with water
and mixture was concentrated under reduced pressure. The resulted
residue was partitioned between EtOAc and water, the separated
organic layer was washed with brine solution, dried over anhydrous
Na.sub.2SO.sub.4, filtered and filtrate was evaporated under
reduced pressure. The resulted crude compound was purified by flash
column chromatography (100-200 silica mesh, eluent was 20% EtOAc in
pet ether) to obtained ethyl
2-(4-chloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)ben-
zo[b]thiophen-3-yl)acetate (600 mg, 43.0% yield) as a colorless
liquid. LCMS (ES) m/z 422.10 [M+H].sup.+.
b) Ethyl
2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)-
oxy)benzo[b]thiophen-3-yl)acetate
##STR00132##
[0500] Tetrakis (164 mg, 0.142 mmol) was added to a degassed
solution of ethyl
2-(4-chloro-6-((2-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)o-
xy)benzo[b]thiophen-3-yl)acetate (600 mg, 1.421 mmol) and
dicyanozinc (167 mg, 1.421 mmol) in DMF (10 mL). The mixture was
further degassed for 10 min and heated to 120.degree. C. for 1 h
under microwave condition. The reaction mixture was filtered
through a pad of Celite.RTM. and filtrate was partitioned between
EtOAc and water. The separated organic layer was washed with brine
solution, dried over anhydrous Na.sub.2SO.sub.4, filtered and
filtrate was evaporated under reduced pressure to get the crude.
The crude was purified by silica gel column chromatography by using
EtOAc in hexane as eluent. The product was eluted at 40% EtOAc-Pet
ether to get ethyl
2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benz-
o[b]thiophen-3-yl)acetate (380 mg, 64.4% yield) as an off-white
solids. .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. 8.06 (d, J=8.0
Hz, 1H), 7.91 (d, J=8.0 Hz, 1H), 7.82 (d, J=2.0 Hz, 1H), 7.53 (s,
1H), 7.17 (d, J=2.0 Hz, 1H), 6.10-6.07 (m, 1H), 4.13 (q, J=7.0 Hz
2H), 4.01 (s, 2H), 3.20-3.13 (m, 1H), 3.07-3.03 (m, 1H), 2.77-2.72
(m, 1H), 2.16-2.12 (m, 1H), 1.20 (t, J=7.5 Hz, 3H). LCMS (ES) m/z
413.25 [M+H].sup.+.
c)
2-(4-Chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)be-
nzo[b]thiophen-3-yl)acetic Acid
##STR00133##
[0502] 3N HCl (10 mL, 30.0 mmol) was added to a stirred solution of
ethyl
2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benz-
o[b]thiophen-3-yl)acetate (0.35 g, 0.848 mmol) in THF (50 mL) at
0.degree. C. The reaction mixture was stirred and heated to
70.degree. C. for 8 h. Evaporated the excess of solvents under
reduced pressure and water was added to the reaction mixture. The
precipitated solid was filtered and dried under vacuum to get crude
material. The resulted crude compound was purified by flash column
chromatography (100-200 silica mesh, eluent was 3% MeOH-DCM to
obtained
2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benz-
o[b]thiophen-3-yl)acetic acid (0.2000 g, 58.8% yield) as an
off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 12.36
(brs, 1H), 8.06 (d, J=7.6 Hz, 1H), 7.91 (d, J=7.6 Hz, 1H), 7.81 (d,
J=2.0 Hz, 1H), 7.50 (s, 1H), 7.17 (d, J=2.0 Hz, 1H), 6.10-6.07 (m,
1H), 4.01 (s, 2H), 3.20-3.13 (m, 1H), 3.07-3.03 (m, 1H), 2.77-2.72
(m, 1H), 2.16-2.12 (m, 1H). LCMS (ES) m/z 385.10 [M+H].sup.+.
d)
2-(6-((2-carbamoyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-4-chlo-
robenzo[b]thiophen-3-yl)acetic acid: (Example 9)
##STR00134##
[0504] H.sub.2O.sub.2 (0.064 mL, 2.079 mmol) was added to a stirred
solution of
2-(4-chloro-6-((2-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)benz-
o[b]thiophen-3-yl)acetic acid (400 mg, 1.039 mmol) in KOH (117 mg,
2.079 mmol) and ethanol (50 mL) at 0.degree. C. The reaction
mixture was stirred at RT for 1 h. The reaction mixture was
concentrated under reduced pressure to get crude. Water (10 mL) was
added and adjusted acidic pH by using 2N citric acid solution and
then filtered the precipitated solid. The solid was washed with
n-pentane to get
2-(6-((2-carbamoyl-6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl)oxy)-4-chloro-
benzo[b]thiophen-3-yl)acetic acid (200 mg, 47.7% yield) as an
off-white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. 12.35
(brs, 1H), 8.06 (s, 1H), 7.97 (d, J=8.0 Hz, 1H), 7.92 (d, J=8.0 Hz,
1H), 7.81 (d, J=2.0 Hz, 1H), 7.64 (s, 1H), 7.49 (s, 1H), 7.16 (d,
J=2.0 Hz, 1H), 6.08-6.06 (m, 1H), 4.01 (s, 2H), 3.20-3.14 (m, 1H),
3.05-3.00 (m, 1H), 2.76-2.72 (m, 1H), 2.17-2.13 (m, 1H). LCMS (ES)
m/z 402.82 [M+H].sup.+. Chiral HPLC: 49.44%: 50.55%.
Analytical SFC Condition
[0505] Column/dimensions: Chiralpak-IG (4.6.times.250 mm),
5.mu.
% CO.sub.2:50.0%
[0506] % Co solvent: 50.0% (100% MeOH) Total Flow: 4.0 g/min
Back Pressure: 100 bar
Temperature: 30.degree. C.
UV: 214 nm
Preparative SFC Condition
[0507] Column/dimensions: Chiralpak-IG (30.times.250 mm), 5.mu.
% CO.sub.2: 50.0%
[0508] % Co solvent: 50.0% (100% MeOH) Total Flow: 90.0 g/min
Back Pressure: 90.0 bar
UV: 214 nm
[0509] Stack time: 15.5 min
Load/Inj: 15.0 mg
[0510] Retention time: Peak 1-11.41 min, Peak 2-14.44 min.
Purity: Peak 1-99.61%, Peak 2-99.07%.
[0511] Solubility: Few drops of H.sub.2O+THF+MeOH Instrument
details: Make/Model: SFC-200-003
Chiral Separation of Examples 9
Examples 9a (First Eluted Enantiomer):
[0512] .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. 12.35 (brs,
1H), 8.06 (s, 1H), 7.97 (d, J=8.0 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H),
7.81 (d, J=2.0 Hz, 1H), 7.64 (s, 1H), 7.49 (s, 1H), 7.16 (d, J=2.0
Hz, 1H), 6.08-6.06 (m, 1H), 4.01 (s, 2H), 3.20-3.14 (m, 1H),
3.05-3.00 (m, 1H), 2.76-2.72 (m, 1H), 2.17-2.13 (m, 1H). LCMS (ES)
m/z 403.22 [M+H].sup.+. Chiral HPLC: 99.61%.
Examples 9b (Second Eluted Enantiomer):
[0513] .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. 12.35 (brs,
1H), 8.06 (s, 1H), 7.97 (d, J=8.0 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H),
7.81 (d, J=2.0 Hz, 1H), 7.64 (s, 1H), 7.49 (s, 1H), 7.16 (d, J=2.0
Hz, 1H), 6.08-6.06 (m, 1H), 4.01 (s, 2H), 3.20-3.14 (m, 1H),
3.05-3.00 (m, 1H), 2.76-2.72 (m, 1H), 2.17-2.13 (m, 1H). LCMS (ES)
m/z 403.28 [M+H].sup.+. Chiral HPLC: 99.07%
Assay Protocol
[0514] Compounds contained herein were evaluated for their ability
to inhibit the activity of GOAT. GOAT activity was assessed using a
time-resolved fluorescence energy transfer (TR-FRET) assay in a
384-well format. His-tag human GOAT enzyme was in the form of a
cell membrane preparation from sf9 cells infected with hGOAT-V5-His
baculovirus. Varying concentrations of test compound with final
DMSO concentration kept to 0.5% were added to membrane solution.
Human GOAT membrane activity was established in a buffer having
final concentration 0.25 mg/mL in 50 mM MOPS, pH7.5; 50 mM KCl; 0.1
mg/mL BSA; 50 .mu.M CHAPS; and 2 mM EDTA. Substrate solution
consisting of biotinylated ghrelin peptide (final concentration 100
nM), octanoyl coA (final concentration 2 .mu.M) and palmitoyl CoA
(final concentration 50 .mu.M) was added to initiate the reaction.
Plates were sealed, centrifuged for 1 minute at 2000 rpm, then
incubated at 30.degree. C. for 80 minutes with gentle shaking on an
Eppendorf mix plate. Reaction termination and detection mix
consisting of chicken anti-active ghrelin antibody (final
concentration of 10 nM), Europium W1024-labeled streptavidin (final
concentration of 4 nM), GOAT anti-chicken Dylight (final
concentration of 12.5 nM), and GS[DAP-oc]-FL-amide inhibitor (final
concentration of 1 .mu.M) was added before further incubation for
40 minutes at 30.degree. C. The plate was then read on an Envision
in HTRF mode with excitation filter UV (TRF) 340 and first emission
filter of APC 665 and a second emission filter of Europium 615.
HTRF readings were acquired as per instrument defined LANCE-DELFIA
protocol with a delay and window times of 50 .mu.s for both; number
of sequential windows: 1; time between flashes: 2000 .mu.s between
each of 100 flashes and 10 flashes for the second detector. The
HTRF ratio was calculated directly by the instrument as the ratio
of 665 window/615 window. Percent inhibition was calculated as
100-(100.times.(U-NC)/(PC-NC)) where U was the unknown value HTRF
ratio (test compound value), NC was the negative control (100%
inhibition value generated from a potent inhibitor), and PC was the
positive control (100% activity generated from 0.5% DMSO vehicle).
IC.sub.50 values were generated in GraphPad Prism (Version 4.03)
using non-linear regression curve fit and sigmoidal dose response
variable slope analysis.
Results
[0515] The exemplified compounds were generally tested according to
the above or an analogous assay and were found to be inhibitors of
GOAT. Specific biological activities tested according to such
assays are listed in the following table as follows (IC.sub.50): A
<50 nM, B: <500 nM, C: <5000 nM. As variability in such
assays is inevitable, repeating the assay run(s) may result in
slightly different IC.sub.50 values.
TABLE-US-00005 hGOAT IC.sub.50 Example (nM) 1a A 1b C 2a A 2b C 3a
B 3b A 4a B 4b A 5a C 5b A 6a A 6b A 7a C 7b A 8 A 9a B 9b A
Evaluation in Animal Models
[0516] The activity of Example 1a was evaluated in vivo in three
preclinical species by assessing the level of reduction of acyl
ghrelin in circulation after treatment.
Acyl Ghrelin Reduction in Mice
[0517] Normal mice were administered various oral doses of Example
1a on a bid basis for two days (4 doses). Food was withdrawn the
evening of the second day after the fourth dose, and then the
animals were administered a final dose the morning of the third
day. Three hours after this fifth and final dose, blood was
collected for ghrelin and acyl ghrelin measurement by ELISA.
Dose-dependent decreases in acyl ghrelin and increases in des-acyl
ghrelin were observed. As seen in FIG. 3, acyl ghrelin reductions
were statistically significant (p<0.05) at both 1 and 10 mg/kg
Example 1a.
Acyl Ghrelin Reduction in Rats
[0518] The same experimental design was conducted in rats
administered 10 mg/kg of Example 1a. The GOAT inhibitor
significantly reduced acyl ghrelin levels (FIG. 4) and increased
des-acyl ghrelin levels (FIG. 5) in rats.
Acyl Ghrelin Reduction in Cynomolgus Monkeys
[0519] A single-dose PK/PD study was conducted in cynomolgus
monkeys after a 10 mg/kg dose of Example 1a. On day zero, after an
overnight fast, three monkeys were administered an oral dose of
vehicle, and blood was collected pre-dose and 1, 3, 8, and 24 hours
later. The 24 hour time point served as the pre-dose measurement
for day 1 when, after an overnight fast, the monkeys were
administered a single oral dose of 10 mg/kg Example 1a. Blood was
collected at various time points (15 minutes, 30 minutes, 1 hour, 3
hours, 8 hours, 24 hours, 48 hours, 96 hours, and 168 hours) with
overnight fasting throughout the study. Levels of acyl ghrelin were
measured using a Millipore metabolic panel that did not include
des-acyl ghrelin. Over the first 24 hours, treatment with Example
1a caused a 51% reduction in the acyl ghrelin AUC relative to
vehicle treatment (FIG. 6).
Acyl Ghrelin Reduction in Mice on a High Fat, High Carb Diet
[0520] Normal mice were acclimated to individual housing for two
weeks on normal chow diet (20% calories from protein, 35% from
carbohydrate, and 45% from fat). All mice except a control group
were then switched to a high fat, high carb (HFHC) diet to cause
obesity. Mice on the HFHC diet were administered 3 mg/kg Rimonabant
once daily for seven days to cause weight loss except for one group
that initiated 10 mg/kg Example 1a immediately. Mice that had been
treated with Rimonabant were then administered various treatments
in the evening for 21 days: vehicle, 3 mg/kg Rimonabant, or 10
mg/kg Example 1a. Mice were fasted overnight and administered a
final treatment in the morning. Three hours later, blood was
collected for plasma acyl ghrelin and des-acyl ghrelin
determination.
[0521] As seen in FIG. 7, by the end of the study, mice fed the
HFHC diet, regardless of other treatment, had lower acyl ghrelin
levels than mice fed normal chow. Continuous treatment with Example
1a and treatment with Example 1a after Rimonabant produced
statistically significant 57% and 61% reductions in acyl ghrelin
levels compared to Rimonabant followed by vehicle. Continuous
treatment of Example 1a produced a 28% reduction in acyl ghrelin
compared to vehicle treatment alone. As seen in FIG. 8, des-acyl
ghrelin levels were statistically significantly elevated by both
Example 1a treatments relative to the Rimonabant followed by
vehicle treatment. Rimonabant treatment alone also led to increased
acyl ghrelin levels, but not as much as that seen for treatment
with Example 1a with or without Rimonabant pretreatment.
* * * * *