Note: Descriptions are shown in the official language in which they were submitted.
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4-(5-CYANO-PYRAZOL-1-YL)-PIPERIDINE DERIVATIVES AS GPR 119 MODULATORS
FIELD OF THE INVENTION
The present invention relates to a new class of cyanopyrazoles, pharmaceutical
compositions containing these compounds, and their use to modulate the
activity of the
G-protein-coupled receptor, GPR119.
BACKGROUND
Diabetes mellitus are disorders in which high levels of blood glucose occur as
a
consequence of abnormal glucose homeostasis. The most common forms of diabetes
mellitus are Type I (also referred to as insulin-dependent diabetes mellitus)
and Type II
diabetes (also referred to as non-insulin-dependent diabetes mellitus). Type
II diabetes,
accounting for roughly 90% of all diabetic cases, is a serious progressive
disease that
results in microvascular complications (including retinopathy, neuropathy and
nephropathy) as well as macrovascular complications (including accelerated
atherosclerosis, coronary heart disease and stroke).
Currently, there is no cure for diabetes. Standard treatments for the disease
are
limited, and focus on controlling blood glucose levels to minimize or delay
complications.
Current treatments target either insulin resistance (metformin,
thiazolidinediones, or
insulin) release from beta cells (sulphonylureas, exanatide). Sulphonylureas
and other
compounds that act via depolarization of the beta cell promote hypoglycemia as
they
stimulate insulin secretion independent of circulating glucose concentrations.
One
approved drug, exanatide, stimulates insulin secretion only in the presence of
high
glucose, but must be injected due to a lack of oral bioavailablity.
Sitagliptin, a dipeptidyl
peptidase IV inhibitor, is a new drug that increases blood levels of incretin
hormones,
which can increase insulin secretion, reduce glucagon secretion and have other
less
well characterized effects. However, sitagliptin and other dipeptidyl
peptidases IV
inhibitors may also influence the tissue levels of other hormones and
peptides, and the
long-term consequences of this broader effect have not been fully
investigated.
In Type II diabetes, muscle, fat and liver cells fail to respond normally to
insulin.
This condition (insulin resistance) may be due to reduced numbers of cellular
insulin
receptors, disruption of cellular signaling pathways, or both. At first, the
beta cells
compensate for insulin resistance by increasing insulin output. Eventually,
however, the
beta cells become unable to produce sufficient insulin to maintain normal
glucose levels
(euglycemia), indicating progression to Type II diabetes.
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In Type II diabetes, fasting hyperglycemia occurs due to insulin resistance
combined with beta cell dysfunction. There are two aspects of beta cell defect
dysfunction: 1) increased basal insulin release (occurring at low, non-
stimulatory
glucose concentrations), which is observed in obese, insulin-resistant pre-
diabetic
stages as well as in Type II diabetes, and 2) in response to a hyperglycemic
challenge,
a failure to increase insulin release above the already elevated basal level,
which does
not occur in pre-diabetic stages and may signal the transition from normo-
glycemic
insulin-resistant states to Type II diabetes. Current therapies to treat the
latter aspect
include inhibitors of the beta-cell ATP-sensitive potassium channel to trigger
the release
of endogenous insulin stores, and administration of exogenous insulin. Neither
achieves
accurate normalization of blood glucose levels and both carry the risk of
eliciting
hypoglycemia.
Thus, there has been great interest in the discovery of agents that function
in a
glucose-dependent manner. Physiological signaling pathways which function in
this way
are well known, including gut peptides GLP-1 and GIP. These hormones signal
via
cognate G-protein coupled receptors to stimulate production of cAMP in
pancreatic
beta-cells. Increased cAMP apparently does not result in stimulation of
insulin release
during the fasting or pre-prandial state. However, a number of biochemical
targets of
cAMP, including the ATP-sensitive potassium channel, voltage-sensitive
potassium
channels and the exocytotic machinery, are modulated such that insulin
secretion due
to postprandial glucose stimulation is significantly enhanced. Therefore,
agonist
modulators of novel, similarly functioning, beta-cell GPCRs, including GPR119,
would
also stimulate the release of endogenous insulin and promote normalization of
glucose
levels in Type II diabetes patients. It has also been shown that increased
cAMP, for
example as a result of GLP-1 stimulation, promotes beta-cell proliferation,
inhibits beta-
cell death and, thus, improves islet mass. This positive effect on beta-cell
mass should
be beneficial in Type II diabetes where insufficient insulin is produced.
It is well known that metabolic diseases have negative effects on other
physiological systems and there is often co-occurrence of multiple disease
states (e.g.,
Type I diabetes, Type ll diabetes, inadequate glucose tolerance, insulin
resistance,
hyperglycemia, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia,
dyslipidemia, obesity or cardiovascular disease in "Syndrome X") or secondary
diseases which occur secondary to diabetes such as kidney disease, and
peripheral
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neuropathy. Thus, treatment of the diabetic condition should be of benefit to
such
interconnected disease states.
SUMMARY OF THE INVENTION
In accordance with the present invention, a new class of GPR 119 modulators
has been discovered. These compounds include:
Isopropyl 4-{5-cyano-4-[(2,4-difluorophenoxy)methyI]-1H-pyrazol-1-
yllpiperidine-1-
carboxylate;
Isopropyl 4-{5-cyano-4-[(2-methylphenoxy)methy1]-1H-pyrazol-1-yllpi peridine-1-
carboxylate;
1-Methylcyclopropyl 4-{5-cyano-4-[(2,5-difluorophenoxy)methyI]-1H-pyrazol-1-
yllpiperidine-1-carboxylate;
1-Methylcyclopropyl 4-{5-cyano-4-[(2,3-difluorophenoxy)methyI]-1H-pyrazol-1-
yllpiperidine-1-carboxylate;
1-Methylcyclopropyl 4-{4-[(4-carbamoy1-2-fluorophenoxy)methyl]-5-cyano-1H-
pyrazol-1-
yllpiperidine-1-carboxylate;
1-Methylcyclopropyl 4-{4-[(4-carbamoylphenoxy)methy1]-5-cyano-1H-pyrazol-1-
yllpiperidine-1-carboxylate;
1-Methylcyclopropyl 4-(5-cyano-4-((4-cyanophenoxy)methyl)-1H-pyrazol-1-
yl)piperidine-
1-carboxylate;
Isopropyl 4-(4-((4-(1H-pyrazol-1-yl)phenoxy)methyl)-5-cyano-1H-pyrazol-1-
yl)piperidine-
1-carboxylate;
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1H-tetrazol-5-yl)phenoxy)methyl)-1H-
pyrazol-1-
yl)piperidine-1-carboxylate and Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate;
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-methyl-1H-tetrazol-5-yl)phenoxy)methyl)-
1H-
pyrazol-1-yl)piperidine-1-carboxylate;
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-methyl-2H-tetrazol-5-yl)phenoxy)methyl)-
1H-
pyrazol-1-yl)piperidine-1-carboxylate;
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-(2-hydroxyethyl)-2H-tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate;
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-(2-hydroxyethyl)-1H-tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate;
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1-Methylcyclopropyl 4-(5-cyano-4-{[2-fluoro-4-(1 -methyl-1 H-tetrazol-5-
yl)phenoxy]methyll-1 H-pyrazol-1-yl)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(5-cyano-4-{[4-(1 -methyl-1 H-tetrazol-5-
yl)phenoxy]methyll-1 H-
pyrazol-1-yl)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(4-((4-carbamoy1-3-fluorophenoxy)methyl)-5-cyano-1 H-
pyrazol-1-
yl)piperidine-1-carboxylate;
Isopropyl 4-(5-cyano-4-{142-fluoro-4-(methylsulfonyl)phenoxy]ethy11-1H-pyrazol-
1-
y1)piperidine-1-carboxylate;
Isopropyl 4-(5-cyano-4-{1-[(2-methylpyridin-3-yl)oxy]ethyll-1 H-pyrazol-1-
yl)piperidine-1-
carboxylate;
Isopropyl 4-(5-cyano-4-{242-fluoro-4-(methylsulfonyl)phenyl]propy11-1H-pyrazol-
1-
yl)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(5-cyano-4-{[(2-methylpyridin-3-yl)oxy]methyll-1 H-
pyrazol-1-
yl)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-{5-cyano-4-[(2,3,6-trifluorophenoxy)methyI]-1 H-pyrazol-
1-
yllpiperidine-1-carboxylate;
Isopropyl 4-{5-cyano-4-[(2,3,6-trifluorophenoxy)methyI]-1H-pyrazol-1-
yllpiperidine-1-
carboxylate;
Isopropyl 4-(5-cyano-4-{[2-fluoro-4-(1-methyl-1 H-imidazol-2-
yl)phenoxy]methyll-1 H-
pyrazol-1-yl)piperidine-1-carboxylate;
Isopropyl 4-(5-cyano-4-{[2-fluoro-4-(1-methyl-1H-imidazol-5-yl)phenoxy]methyll-
1H-
pyrazol-1-y1)piperidine-1-carboxylate;
Isopropyl 445-cyano-4-({[2-methyl-6-(1 H-1,2,4-triazol-1-yl)pyridin-3-
yl]oxylmethyl)-1H-
pyrazol-1-yl]piperidine-1-carboxylate;
Isopropyl 445-cyano-4-({[2-methyl-6-(1 H-1,2,4-triazol-1-yl)pyridin-3-
yl]aminolmethyly
1 H-pyrazol-1-yl]piperidine-1-carboxylate;
Isopropyl 4[5-cyano-4-({[2-methyl-6-(methylsulfonyl)pyridin-3-yl]aminolmethyl)-
1 H-
pyrazol-1-yl]piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(5-cyano-4-{[4-(1H-tetrazol-1-yl)phenoxy]rnethyll-1 H-
pyrazol-1-
yl)piperidine-1-carboxylate;
141-(5-Ethylpyrimidin-2-yl)piperidin-4-y1]-4-{[4-(1 H-tetrazol-1-
yl)phenoxy]methyll-1 H-
pyrazole-5-carbonitrile;
Isopropyl 4-{5-cyano-4-[(3-cyanophenoxy)methy1]-1 H-pyrazol-1-yllpiperidine-1-
carboxylate;
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Isopropyl 4-{5-cyano-4-[(4-cyano-3-methylphenoxy)methy1]-1 H-pyrazol-1-
yllpiperidine-
1-carboxylate;
Isopropyl 4-{5-cyano-4-[(4-cyanophenoxy)methy1]-1H-pyrazol-1-yllpiperidine-1-
carboxylate;
4-[(4-Cyano-2-fluorophenoxy)nethyl]-141-(5-ethylpyrimidin-2-y1)piperidin-4-y1]-
1H-
pyrazole-5-carbonitrile;
tert-Butyl 4-{5-cyano-4-[(4-cyano-2-fluorophenoxy)methy1]-1H-pyrazol-1-
yllpiperidine-1-
carboxylate;
Isopropyl 4-{5-cyano-4-[(2-cyano-4-fluorophenoxy)methy1]-1H-pyrazol-1-
yllpiperidine-1-
carboxylate;
Isopropyl 4-(5-cyano-4-{[4-(dirnethylcarbarnoy1)-2-fluorophenoxy]rnethyll-1H-
pyrazol-1-
y1)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(5-cyano-4-{[4-(dirnethylcarbarnoy1)-2-
fluorophenoxy]rnethyll-1H-
pyrazol-1-y1)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(5-cyano-4-{[2-fluoro-4-
(rnethylcarbarnoyl)phenoxy]rnethyll-1 H-
pyrazol-1-yl)piperidine-1-carboxylate;
4-({5-Cyano-141-(5-ethylpyrimidin-2-yl)piperidin-4-y1]-1H-pyrazol-4-
yllmethoxy)-3-fluoro-
N,N-dimethylbenzamide;
1-Methylcyclopropyl 4-{5-cyano-4-[(4-cyano-2-fluorophenoxy)methy1]-1H-pyrazol-
1-
yllpiperidine-1-carboxylate;
tert-Butyl (3S,4S)-4-(5-cyano-4-{[2-fluoro-4-
(rnethylcarbarnoyl)phenoxy]rnethyll-1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate;
tert-Butyl (3R,4S)-4-(5-cyano-4-{[2-fluoro-4-
(rnethylcarbarnoyl)phenoxy]rnethyll-1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate;
1-Methylcyclopropyl (3S,4S)-4-(5-cyano-4-{[2-fluoro-4-
(rnethylcarbarnoyl)phenoxy]rnethyll-1 H-pyrazol-1-y1)-3-fluoropiperidine-1-
carboxylate;
1-Methylcyclopropyl (3R,4R)-4-(5-cyano-4-{[2-fluoro-4-
(rnethylcarbarnoyl)phenoxy]rnethyll-1 H-pyrazol-1-y1)-3-fluoropiperidine-1-
carboxylate;
tert-Butyl (3S,4S)-4-(5-cyano-4-{[(2-rnethylpyridin-3-yl)oxy]rnethyll-1H-
pyrazol-1-y1)-3-
fluoropiperidine-1-carboxylate;
tert-Butyl (3S,4R)-4-(5-cyano-4-{[(2-rnethylpyridin-3-yl)oxy]rnethyll-1H-
pyrazol-1-y1)-3-
fluoropiperidine-1-carboxylate;
1-Methylcyclopropyl (3S,4R)-4-(5-cyano-4-{[(2-rnethylpyridin-3-yl)oxy]rnethyll-
1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate;
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1-Methylcyclopropyl (3S,4R)-4-(5-cyano-4-{[(2-methylpyridin-3-yl)oxy]methyll-
1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate;
1-Methylcyclopropyl (3 R,4S)-4-(5-cyano-4-{[(2-methylpyridin-3-yl)oxy]methyll-
1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate;
tert-Butyl 4-(5-cyano-4-{[4-(1 H-1,2, 3-triazol-1-yl)phenoxy]rnethyll-1 H-
pyrazol-1-
yl)piperidine-1-carboxylate;
tert-Butyl 4-(5-cyano-4-{[4-(2H-1,2,3-triazol-2-yl)phenoxy]methyll-1H-pyrazol-
1-
y1)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(4-((4-(1H-1,2,3-triazol-1-yl)phenoxy)methyl)-5-cyano-1H-
pyrazol-1-yl)piperidine-1-carboxylate;
1-Methylcyclopropyl 4-(4-((4-(2H-1,2,3-triazol-2-yl)phenoxy)methyl)-5-cyano-1H-
pyrazol-1-yl)piperidine-1-carboxylate;
tert-Butyl 445-cyano-4-({[1-(methylsulfonyl)piperidin-4-yl]oxylmethyl)-1H-
pyrazol-1-
yl]piperidine-1-carboxylate;
tert-Butyl 445-cyano-4-({2-fluoro-4-[(2-
hydroxyethyl)(methyl)carbamoyl]phenoxylmethyl)-1H-pyrazol-1-yl]pi peridine-1-
carboxylate;
tert-Butyl 445-cyano-4-({2-fluoro-4-[(3-hydroxypyrrolidin-1-
yl)carbonyl]phenoxylmethyl)-
1H-pyrazol-1-yl]piperidine-1-carboxylate;
tert-Butyl 4-(4-{[4-(azetidin-1-ylcarbony1)-2-fluorophenoxy]methyll-5-cyano-1H-
pyrazol-
1-y1)piperidine-1-carboxylate;
1-Methylcyclopropyl 445-cyano-4-({[1-(methylsulfonyl)piperidin-4-
yl]oxylmethyl)-1H-
pyrazol-1-yl]piperidine-1-carboxylate;
1-methylcyclopropyl 4-(5-cyano-4-((2-fluoro-4-(1H-1,2, 3-triazol-1-
yl)phenoxy)methyly
1H-pyrazol-1-yl)piperidine-1-carboxylate;
isopropyl 4-(5-cyano-4-((2-fluoro-4-(1 H-1,2, 3-triazol-1-yl)phenoxy)methyl)-1
H-pyrazol-1-
yl)piperidine-1-carboxylate;
1-(1-(5-ethylpyrimidin-2-yl)piperidin-4-y1)-4-((2-fluoro-4-(1H-1,2,3-triazol-1-
yl)phenoxy)methyl)-1H-pyrazole-5-carbonitrile;
isopropyl 4-(4-((4-(1H-1,2,3-triazol-1-yl)phenoxy)methyl)-5-cyano-1 H-pyrazol-
1-
yl)piperidine-1-carboxylate;
4-((4-(1 H-1,2,3-triazol-1-yl)phenoxy)methyl)-1-(1-(5-ethylpyrim idin-2-yl)pi
peridin-4-y1)-
1H-pyrazole-5-carbonitrile;
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isopropyl 4-(4-((4-(2H-1,2,3-triazol-2-yl)phenoxy)methyl)-5-cyano-1H-pyrazol-1-
yl)piperidine-1-carboxylate;
4-((4-(2H-1,2,3-triazol-2-yl)phenoxy)methyl)-1-(1-(5-ethylpyrim id in-2-yl)pi
perid in-4-y1)-
1H-pyrazole-5-carbonitrile;
1-methylcyclopropyl 4-(5-cyano-4-((2-fluoro-4-(2H-1,2,3-triazol-2-
yl)phenoxy)methyl)-
1H-pyrazol-1-yl)piperidine-1-carboxylate;
isopropyl 4-(5-cyano-4-((2-fluoro-4-(2H-1,2,3-triazol-2-yl)phenoxy)methyl)-1H-
pyrazol-1-
yl)piperidine-1-carboxylate;
1-(1-(5-ethylpyrim idin-2-yl)pi peridin-4-y1)-4-((2-fluoro-4-(2H-1,2,3-triazol-
2-
yl)phenoxy)methyl)-1H-pyrazole-5-carbonitrile;
1-methylcyclopropyl 4-(4-((5-(1H-1,2, 3-triazol-1-yl)pyridin-2-yloxy)methyl)-5-
cyano-1H-
pyrazol-1-yl)piperid me-1-carboxylate;
isopropyl 4-(4-((5-(1H-1,2,3-triazol-1-yl)pyridin-2-yloxy)methyl)-5-cyano-1H-
pyrazol-1-
yl)piperidine-1-carboxylate;
1-methylcyclopropyl 4-(5-cyano-4-((3-fluoro-4-(1H-tetrazol-1-
yl)phenoxy)methyl)-1H-
pyrazol-1-yl)piperid me-1-carboxylate;
isopropyl 4-(5-cyano-4-((3-fluoro-4-(1H-tetrazol-1-yl)phenoxy)methyl)-1H-
pyrazol-1-
yl)piperidine-1-carboxylate;
1-(1-(5-ethylpyrim idin-2-yl)pi peridin-4-y1)-4-((3-fluoro-4-(1H-tetrazol-1-
yl)phenoxy)methyl)-1H-pyrazole-5-carbonitrile;
4-((5-(1 H-1,2,3-triazol-1-yl)pyridin-2-yloxy)methyl)-1-(1-(5-ethylpyrimidin-2-
yl)piperidin-
4-y1)-1 H-pyrazole-5-carbonitrile;
1-(1-(5-ethylpyrimidin-2-yl)piperidin-4-y1)-4-((2-methy1-6-(1H-1,2,3-triazol-1-
yl)pyridin-3-
yloxy)methyl)-1H-pyrazole-5-carbonitrile;
isopropyl 4-(5-cyano-4-((2-methy1-6-(1H-1,2,3-triazol-1-yl)pyridin-3-
yloxy)methyl)-1H-
pyrazol-1-yl)piperidine-1-carboxylate;
1-methylcyclopropyl 4-(5-cyano-4-((2-methy1-6-(1H-1,2,3-triazol-1-yl)pyridin-3-
yloxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate;
1-(1-(5-ethylpyrim idin-2-yl)pi peridin-4-y1)-4-((2-fluoro-4-(1-methy1-1 H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazole-5-carbonitrile;
1-methylcyclopropyl 4-(4-((4-(azetidine-1-carbony1)-2-fluorophenoxy)methyl)-5-
cyano-
1H-pyrazol-1-y1)piperidine-1-carboxylate;
isopropyl 4-(4-((4-(azetidine-1-carbony1)-2-fluorophenoxy)methyl)-5-cyano-1H-
pyrazol-
1-y1)piperidine-1-carboxylate; and
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4-((4-(azetidine-1-carbony1)-2-fluorophenoxy)methyl)-1-(1-(5-ethylpyrimidin-2-
yl)piperidin-4-yI)-1H-pyrazole-5-carbonitrile;
or a pharmaceutically acceptable salt thereof.
These compounds modulate the activity of the G-protein-coupled receptor. More
specifically the compounds modulate GPR119. As such, said compounds are useful
for
the treatment of diseases, such as diabetes, in which the activity of GPR119
contributes
to the pathology or symptoms of the disease. Examples of such conditions
include
hyperlipidemia, Type I diabetes mellitus, Type!! diabetes mellitus, idiopathic
Type I
diabetes (Type lb), latent autoimmune diabetes in adults (LADA), early-onset
Type 2
diabetes (EOD), youth-onset atypical diabetes (YOAD), maturity onset diabetes
of the
young (MODY), malnutrition-related diabetes, gestational diabetes, coronary
heart
disease, ischemic stroke, restenosis after angioplasty, peripheral vascular
disease,
intermittent claudication, myocardial infarction (e.g. necrosis and
apoptosis),
dyslipidemia, post-prandial lipemia, conditions of impaired glucose tolerance
(IGT),
conditions of impaired fasting plasma glucose, metabolic acidosis, ketosis,
arthritis,
obesity, osteoporosis, hypertension, congestive heart failure, left
ventricular hypertrophy,
peripheral arterial disease, diabetic retinopathy, macular degeneration,
cataract,
diabetic nephropathy, glomerulosclerosis, chronic renal failure, diabetic
neuropathy,
metabolic syndrome, syndrome X, premenstrual syndrome, coronary heart disease,
angina pectoris, thrombosis, atherosclerosis, transient ischemic attacks,
stroke,
vascular restenosis, hyperglycemia, hyperinsulinemia, hyperlipidemia,
hypertrygliceridemia, insulin resistance, impaired glucose metabolism,
conditions of
impaired glucose tolerance, conditions of impaired fasting plasma glucose,
obesity,
erectile dysfunction, skin and connective tissue disorders, foot ulcerations
and
ulcerative colitis, endothelial dysfunction and impaired vascular compliance.
The
compounds may be used to treat neurological disorders such as Alzheimer's
disease,
schizophrenia, and impaired cognition. The compounds will also be beneficial
in
gastrointestinal illnesses such as inflammatory bowel disease, ulcerative
colitis, Crohn's
disease, irritable bowel syndrome, etc. As noted above, the compounds may also
be
used to stimulate weight loss in obese patients, especially those afflicted
with diabetes.
A further embodiment of the invention is directed to pharmaceutical
compositions
containing a compound of this invention. Such formulations will typically
contain a
compound of this invention in admixture with at least one pharmaceutically
acceptable
excipient. Such formulations may also contain at least one additional
pharmaceutical
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agent. Examples of such agents include anti-obesity agents and/or anti-
diabetic agents.
Additional aspects of the invention relate to the use of the compounds of this
invention
in the preparation of medicaments for the treatment of diabetes and related
conditions
as described herein.
It is to be understood that both the foregoing summary and the following
detailed
description are exemplary and explanatory only and are not restrictive of the
invention,
as claimed.
DETAILED DESCRIPTION OF THE INVENTION
The present invention may be understood even more readily by reference to the
following detailed description of exemplary embodiments of the invention and
the
examples included therein.
It is to be understood that this invention is not limited to specific
synthetic
methods of making that may, of course, vary. It is also to be understood that
the
terminology used herein is for the purpose of describing particular
embodiments only
and is not intended to be limiting. The plural and singular should be treated
as
interchangeable, other than the indication of number:
a. "halogen" refers to a chlorine, fluorine, iodine, or bromine atom;
b. "C1- C4 alkyl" refers to a branched or straight chained alkyl group
containing
from 1 to 5 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, etc.;
c. "C1- C4 alkoxy" refers to a straight or branched chain alkoxy group
containing
from 1 to 4 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-
butoxy, isobutoxy, etc.;
d. "C3-C6 cycloalkyl" refers to a nonaromatic ring that is fully hydrogenated
and
exists as a single ring. Examples of such carbocyclic rings include
cyclopropyl,
cyclobutyl, cyclopentyl, and cyclohexyl;
e. "C1- C4 haloalkyl" refers to a straight or branched chain alkyl group
containing
from 1 to 4 carbon atoms, substituted with one or more halogen atoms;
f. "C1- C4 haloalkoxy" refers to a straight or branched chain alkoxy group
containing
from 1 to 4 carbon atoms, substituted with one or more halogen atoms;
g. "5 to 10 membered heteroaryl" means a carbocyclic aromatic system having a
total of 5 to 10 ring atoms and containing one, two, three or four heteroatoms
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selected independently from oxygen, nitrogen and sulfur and having one, two or
three rings wherein such rings may be fused. The term "fused" means that a
second ring is present (ie, attached or formed) by having two adjacent atoms
in
common (ie, shared) with the first ring. The term "fused" is equivalent to the
term
"condensed". The term "heteroaryl" embraces aromatic radicals such as
pyridine,
pyridazine, pyrazine, pyrimidine, imidazo[1,2-a]pyridine, imidazo[1,5-
a]pyridine,
[1,2,4]triazolo[4,3-a]pyridine, [1,2,4]triazolo[4,3-b]pyridazine,
[1,2,4]triazolo[4,3-
a]pyrimidine, and [1,2,4]triazolo[1,5-a]pyridine;
h. "therapeutically effective amount" means an amount of a compound of
the
present invention that (i) treats or prevents the particular disease,
condition, or
disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of
the
particular disease, condition, or disorder, or (iii) prevents or delays the
onset of
one or more symptoms of the particular disease, condition, or disorder
described
herein;
i. "patient" refers to warm blooded animals such as, for example, guinea pigs,
mice,
rats, gerbils, cats, rabbits, dogs, monkeys, chimpanzees, and humans;
j. "treat" embraces both preventative, i.e., prophylactic, and
palliative treatment, i.e.,
relieve, alleviate, or slow the progression of the patient's disease (or
condition) or
any tissue damage associated with the disease;
k. the terms "modulated", "modulating", or "modulate(s)", as used herein,
unless
otherwise indicated, refers to the activation of the G-protein-coupled
receptor
GPR119 with compounds of the present invention;
I. "pharmaceutically acceptable" indicates that the substance or
composition must
be compatible chemically and/or toxicologically, with the other ingredients
comprising a formulation, and/or the mammal being treated therewith.
m. "salts" is intended to refer to pharmaceutically acceptable salts and to
salts
suitable for use in industrial processes, such as the preparation of the
compound.
n. "pharmaceutically acceptable salts" is intended to refer to either
pharmaceutically
acceptable acid addition salts" or "pharmaceutically acceptable basic addition
salts" depending upon the actual structure of the compound.
o. "pharmaceutically acceptable acid addition salts" is intended to apply to
any non-
toxic organic or inorganic acid addition salt of the base compounds or any of
its
intermediates. Illustrative inorganic acids which form suitable salts include
hydrochloric, hydrobromic, sulphuric, and phosphoric acid and acid metal salts
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such as sodium monohydrogen orthophosphate, and potassium hydrogen sulfate.
Illustrative organic acids, which form suitable salts include the mono-, di-,
and
tricarboxylic acids. Illustrative of such acids are for example, acetic,
glycolic,
lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric,
citric, ascorbic,
maleic, hydroxymaleic, benzoic, hydroxy-benzoic, phenylacetic, cinnamic,
salicylic, 2-phenoxybenzoic, p-toluenesulfonic acid, and sulfonic acids such
as
methane sulfonic acid and 2-hydroxyethane sulfonic acid. Such salts can exist
in
either a hydrated or substantially anhydrous form. In general, the acid
addition
salts of these compounds are soluble in water and various hydrophilic organic
solvents.
p. "pharmaceutically acceptable basic addition salts" is intended to apply to
any
non-toxic organic or inorganic basic addition salts of the compounds or any of
its
intermediates. Illustrative bases which form suitable salts include alkali
metal or
alkaline-earth metal hydroxides such as sodium, potassium, calcium, magnesium,
or barium hydroxides; ammonia, and aliphatic, alicyclic, or aromatic organic
amines such as methylamine, dimethylamine, trimethylamine, and picoline.
q. "isomer" means "stereoisomer" and "geometric isomer" as defined below.
"Stereoisomer" refers to compounds that possess one or more chiral centers and
each center may exist in the R or S configuration. Stereoisomers includes all
diastereomeric, enantiomeric and epimeric forms as well as racemates and
mixtures thereof. "Geometric isomer" refers to compounds that may exist in
cis,
trans, anti, syn, entgegen (E), and zusammen (Z) forms as well as mixtures
thereof.
Certain of the compounds of this invention may exist as geometric isomers. The
compounds may possess one or more asymmetric centers, thus existing as two, or
more, stereoisomeric forms. The present invention includes all the individual
stereoisomers and geometric isomers of the compounds of this invention and
mixtures
thereof. Individual enantiomers can be obtained by chiral separation or using
the
relevant enantiomer in the synthesis. As noted above, some of the compounds
exist as
isomers. These isomeric mixtures can be separated into their individual
isomers on the
basis of their physical chemical differences by methods well known to those
skilled in
the art, such as by chromatography and/or fractional crystallization.
Enantiomers can
be separated by converting the enantiomeric mixture into a diastereomeric
mixture by
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reaction with an appropriate optically active compound (e.g., chiral auxiliary
such as a
chiral alcohol or Mosher's acid chloride), separating the diastereoisomers and
converting (e.g., hydrolyzing) the individual diastereoisomers to the
corresponding pure
enantiomers. Enantiomers can also be separated by use of a chiral HPLC column.
Alternatively, the specific stereoisomers may be synthesized by using an
optically active
starting material, by asymmetric synthesis using optically active reagents,
substrates,
catalysts or solvents, or by converting one stereoisomer into the other by
asymmetric
transformation.
The present invention also embraces isotopically-labeled compounds of the
present invention which are identical to those recited herein, but for the
fact that one or
more atoms are replaced by an atom having an atomic mass or mass number
different
from the atomic mass or mass number usually found in nature. Examples of
isotopes
that can be incorporated into compounds of the invention include isotopes of
hydrogen,
carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine,
such as 2H,
3H, 110, 130, 140, 13N, 15N, 150, 170, 180, 31F, 32F, 35s, 18F, 1231, 1251 and
36u,-.1,
respectively.
Certain isotopically-labeled compounds of the present invention (e.g., those
labeled with 3H and 14C) are useful in compound and/or substrate tissue
distribution
assays. Certain isotopically labeled ligands including tritium, 14C, 355 and
1251 could be
useful in radioligand binding assays. Tritiated (i.e., 3H) and carbon-14
(i.e., 14C)
isotopes are particularly preferred for their ease of preparation and
detectability.
Further, substitution with heavier isotopes such as deuterium (i.e., 2H) may
afford
certain therapeutic advantages resulting from greater metabolic stability
(e.g., increased
in vivo half-life or reduced dosage requirements) and hence may be preferred
in some
circumstances. Positron emitting isotopes such as 150, 13N, k_, ii¨,
and 18F are useful for
positron emission tomography (PET) studies to examine receptor occupancy.
Isotopically labeled compounds of the present invention can generally be
prepared by
following procedures analogous to those disclosed in the Schemes and/or in the
Examples herein below, by substituting an isotopically labeled reagent for a
non-
isotopically labeled reagent.
Certain compounds of the present invention may exist in more than one crystal
form (generally referred to as "polymorphs"). Polymorphs may be prepared by
crystallization under various conditions, for example, using different
solvents or different
solvent mixtures for recrystallization; crystallization at different
temperatures; and/or
various modes of cooling, ranging from very fast to very slow cooling during
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crystallization. Polymorphs may also be obtained by heating or melting the
compound
of the present invention followed by gradual or fast cooling. The presence of
polymorphs may be determined by solid probe NMR spectroscopy, IR spectroscopy,
differential scanning calorimetry, powder X-ray diffraction or such other
techniques.
In addition, the compounds of the present invention can exist in unsolvated as
well as solvated forms with pharmaceutically acceptable solvents such as
water,
ethanol, and the like. In general, the solvated forms are considered
equivalent to the
unsolvated forms for the purposes of the present invention. The compounds may
also
exist in one or more crystalline states, i.e. as co-crystals, polymorphs, or
they may exist
as amorphous solids. All such forms are encompassed by the invention and
claims.
In an embodiment in the composition of this invention, the composition further
includes at least one additional pharmaceutical agent selected from the group
consisting of an anti-obesity agent and an anti-diabetic agent. Example anti-
obesity
agents include dirlotapide, mitratapide, implitapide, R56918 (CAS No. 403987),
CAS
No. 913541-47-6, lorcaserin, cetilistat, PYY3_36, naltrexone, oleoyl-estrone,
obinepitide,
pramlintide, tesofensine, leptin, liraglutide, bromocriptine, orlistat,
exenatide, AOD-9604
(CAS No. 221231-10-3) and sibutramine. Example anti-diabetic agents include
metformin, acetohexamide, chlorpropamide, diabinese, glibenclamide, glipizide,
glyburide, glimepiride, gliclazide, glipentide, gliquidone, glisolamide,
tolazamide,
tolbutamide, tendamistat, trestatin, acarbose, adiposine, camiglibose,
emiglitate,
miglitol, voglibose, pradimicin-Q, salbostatin, balaglitazone, ciglitazone,
darglitazone,
englitazone, isaglitazone, pioglitazone, rosiglitazone, troglitazone, exendin-
3, exendin-4,
trodusquemine, reservatrol, hyrtiosal extract, sitagliptin, vildagliptin,
alogliptin and
saxagliptin.
In another embodiment of a method of this invention, the compounds or
compositions of this invention may be administered in an effective amount for
treating a
condition selected from the group consisting of hyperlipidemia, Type I
diabetes, Type II
diabetes mellitus, idiopathic Type I diabetes (Type lb), latent autoimmune
diabetes in
adults (LADA), early-onset Type 2 diabetes (EOD), youth-onset atypical
diabetes
(YOAD), maturity onset diabetes of the young (MODY), malnutrition-related
diabetes,
gestational diabetes, coronary heart disease, ischemic stroke, restenosis
after
angioplasty, peripheral vascular disease, intermittent claudication,
myocardial infarction
(e.g. necrosis and apoptosis), dyslipidemia, post-prandial lipemia, conditions
of impaired
glucose tolerance (IGT), conditions of impaired fasting plasma glucose,
metabolic
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acidosis, ketosis, arthritis, obesity, osteoporosis, hypertension, congestive
heart failure,
left ventricular hypertrophy, peripheral arterial disease, diabetic
retinopathy, macular
degeneration, cataract, diabetic nephropathy, glomerulosclerosis, chronic
renal failure,
diabetic neuropathy, metabolic syndrome, syndrome X, premenstrual syndrome,
coronary heart disease, angina pectoris, thrombosis, atherosclerosis,
myocardial
infarction, transient ischemic attacks, stroke, vascular restenosis,
hyperglycemia,
hyperinsulinemia, hyperlipidemia, hypertrygliceridemia, insulin resistance,
impaired
glucose metabolism, conditions of impaired glucose tolerance, conditions of
impaired
fasting plasma glucose, obesity, erectile dysfunction, skin and connective
tissue
disorders, foot ulcerations and ulcerative colitis, endothelial dysfunction
and impaired
vascular compliance, hyper apo B lipoproteinemia, Alzheimer's disease,
schizophrenia,
impaired cognition, inflammatory bowel disease, ulcerative colitis, Crohn's
disease, and
irritable bowel syndrome.
In a further embodiment, the method further includes administering a second
composition comprising at least one additional pharmaceutical agent selected
from the
group consisting of an anti-obesity agent and an anti-diabetic agent, and at
least one
pharmaceutically acceptable excipient. This method may be used for admistering
the
compositions simultaneously or sequentially and in any order.
In yet another embodiment, the compounds of this invention are useful in the
manufacture of a medicament for treating a disease, condition or disorder that
modulates the activity of G-protein-coupled receptor GPR119. Furthermore, the
compounds are useful in the preparation of a medicament for the treatment of
diabetes
or a morbidity associated with said diabetes.
Synthesis
For illustrative purposes, the reaction schemes depicted below provide
potential
routes for synthesizing the compounds of the present invention as well as key
intermediates. For a more detailed description of the individual reaction
steps, see the
Examples section below. Those skilled in the art will appreciate that other
synthetic
routes may be used to synthesize the inventive compounds. Although specific
starting
materials and reagents are depicted in the schemes and discussed below, other
starting
materials and reagents can be easily substituted to provide a variety of
derivatives
and/or reaction conditions. In addition, many of the compounds prepared by the
14
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methods described below can be further modified in light of this disclosure
using
conventional chemistry well known to those skilled in the art.
Compounds of the invention may be synthesized by synthetic routes that include
processes analogous to those well-known in the chemical arts, particularly in
light of the
description contained herein. The starting materials are generally available
from
commercial sources such as Aldrich Chemicals (Milwaukee, WI) or are readily
prepared
using methods known to those skilled in the art (e.g., prepared by methods
generally
described in Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis,
v. 1-19,
Wiley, New York (1967-1999 ed.), or Bei!steins Handbuch der organischen
Chemie, 4,
Aufl. ed. Springer-Verlag, Berlin, including supplements (also available via
the Bei!stein
online database).
The compounds of this invention can be prepared using methods analogously
known in the art for the production of ethers. The reader's attention is
directed to texts
such as: 1) Hughes, D. L.; Organic Reactions 1992, 42 Hoboken, NJ, United
States;
2) Tikad, A.; Routier, S.; Akssira, M.; Leger, J.-M.I; Jarry, C.; Guillaumet,
G. Synlett
2006, 12, 1938-42; and 3) Loksha, Y. M.; Globisch, D.; Pedersen, E. B.; La
CoIla, P.;
CoIlu, G.; Loddo, R. J. Het. Chem. 2008, 45, 1161-6 which describe such
reactions
in greater detail.
Scheme 1
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0 0
0
NHNH2 -\o---- -\ \\
R20,,,,,, -e.Y.L0 0
CN B \\ Step 2 ,N
______________________ ...- Br
N H2N N-N ____________________ N
Step 1 R2,a,....}..õ... R2z...c,L.,
A
õ--
C N D
N 1
Z
Step 31
XµO---\ HO\
HO) \\
\\N ,\\N
NC N-- Br'N
NC "N Step 4
R2.?õ.õ.õ1) X-OH
.., __________________________________ R2z..}.. j
Step 5 .-
N Th\1
Zi G Y F 1
Z E
Z
I Step 6
R1
Xs
HO OHC
-- \\N R5
NC N-- Step 7 \\N Step 9
NC N= NC N-N
..t _________________________________________ -)p..
R2ze,...,
R2z,..),,
R2z,....1,,...
N--- ---
1 Th\1 Th\1
Z J I H
ZZ L
X-OH I
Step 10
Step 8
X R1 R1 X R1
0
sO) \\ \\
NC N-N NC N-N Step 11 R5,N \\
_________________________________________________________________ ..- NC N-N
R2..,,,L. j R2.......õ,,,c R2zicf,.j
õ--
N N
K M Y N
Z
Scheme 1 may be used to prepare compounds of Formula N wherein
16
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(R3) (R3)nµ
/\ \N
1 I
r.CH3
vw
Xis I Or I ,
Z is ¨C(0)-0-R6 or pyrimidine substituted with C1-C4 alkyl, CF3, halogen,
cyano,
C3-C6 cycloalkyl or C3-C6 cycloalkyl wherein one carbon atom of said
cycloalkyl moiety
may optionally be substituted with methyl or ethyl;
m is 1, 2, or 3;
n is 0, 1 or 2;
R1 is hydrogen, C1-C4 alkyl, or C3-C6 cycloalkyl;
R2a is hydrogen, fluoro or C1-C4 alkyl;
each R3 is individually selected from the group consisting of: hydroxy,
halogen,
cyano, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, -S02-R7, -
P(0)(0R8)(0R9), -C(0)-NR8R9 , -N(CH3)-00-0-(Ci-C4) alkyl, -NH-00-0-(Ci-C4)
alkyl,-
NH-00-(C1-C4)alkyl, -N(CH3)-00-(C1-C4) alkyl, -NH-(CH2)2-0H and a 5 to 6-
membered
heteroaryl group containing 1, 2, 3 or 4 heteroatoms each independently
selected from
oxygen, nitrogen and sulfur, wherein a carbon atom on said heteroaryl group is
optionally substituted with R4a or a nitrogen atom on said heteroaryl group is
optionally
substituted with R4b;
R4a is hydrogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, or halogen,
wherein
said alkyl is optionally substituted with hydroxyl or C1-C4 alkoxy;
Rzib is hydrogen, C1-C4 alkyl, -CH2-C1-C3 haloalkyl, -C2-C4 alkyl-OH or -CH2-
C1-C4
alkoxy;
R5 is hydrogen or when R1 is hydrogen then R5 is hydrogen or C1-C4 alkyl;
R6 is C1-C4 alkyl or C3-C6 cycloalkyl wherein one carbon atom of said
cycloalkyl
moiety may optionally be substituted with methyl or ethyl;
R7 is represented by C1-C4 alkyl, C3-C6 cycloalkyl, NH2, or -(CH2)2-0H;
R8 is represented by hydrogen or C1-C4 alkyl; and
R9 is represented by hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, -(CH2)2-0H, -
(CH2)2-0-CH3, -(CH2)3-0H, -(CH2)3-0-CH3, 3-oxetanyl, or 3-hydroxycyclobutyl;
or when R3 is -C(0)-NR8R9, R8 and R9 can be taken together with the nitrogen
atom to which they are attached to form an azetidine, a pyrrolidine, a
piperidine or a
morpholine ring.
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In Step 1, compounds of Formula C can be prepared via a condensation reaction
of compounds of Formula A and the commercial compound B (Sigma-Aldrich) in a
diverse array of solvents including but not limited to ethanol, toluene and
acetonitrile at
temperatures ranging from 22 C to 130 C depending upon the solvent utilized
fora
period of 1 to 72 hours. In cases where compounds of Formula A are hydrogen
chloride
or trifluoroacetic acid salts, base modifiers such as sodium acetate or sodium
bicarbonate may be added in one to three equivalents to neutralize the salts.
The
reaction may be conducted in polar protic solvents such as methanol and
ethanol at
temperatures ranging from 22 C to 85 C. Typical conditions for this
transformation
include the use of 3 equivalents of sodium acetate in ethanol heated at 85 C
for 3 hours.
Compounds of Formula A can be prepared via a four-step procedure starting with
substituted or unsubstituted 4-piperidinone hydrochloride salts (J. Med. Chem.
2004, 47,
2180). First these salts are treated with an appropriate alkyl chloroformate
or bis(alkyl)
dicarbonate in the presence of excess base to form the corresponding alkyl
carbamate.
The ketone group is then condensed with tert-butoxycarbonyl hydrazide to form
the
corresponding N-(tert-butoxy)carbonyl (BOC) protected hydrazone derivative.
This is
subsequently reduced to the corresponding BOC protected hydrazine derivative
using
reducing agents such as sodium cyanoborohydride or sodium
triacetoxyborohydride.
Finally, the N-(tert-butoxy)carbonyl group is cleaved under acidic conditions
such as
trifluoroacetic acid or hydrochloric acid to give compounds of Formula A,
which are
typically isolated and used as the corresponding salts (e.g., dihydrochloride
salt).
In Step 2, compounds of Formula D may be prepared from compounds of
Formula C via the formation of intermediate diazonium salts via the Sandmeyer
reaction
(Comp. Org. Synth., 1991, 6, 203) These salts may be prepared via
diazotization of
compounds of Formula C with sodium nitrite and aqueous acids such as
hydrochloric,
hydrobromic, sulfuric, nitric, phosphoric and acetic alone or in combinations.
This
reaction is typically carried out in water at 0 C to 100 C. Alternatively,
anhydrous
conditions using alkyl nitrites such as tert-butylnitrite with solvents such
as acetonitrile
may be utilized (J. Med. Chem. 2006, 49, 1562) at temperatures ranging from 0
C to
95 C. These diazonium intermediates are then allowed to react with copper
salts such
as copper(II) bromide, copper(I) bromide or with tribromomethane to form
compounds
of Formula D. Typical conditions for this transformation include the use of
tert-butylnitrite,
copper(II) bromide in acetontrile at 65 C for 30 minutes.
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In Step 3, compounds of Formula E may be prepared from compounds of
Formula D via the use of reducing agents such as lithium aluminum hydride,
sodium
borohydride, lithium borohydride, borane-dimethylsulfide, borane-
tetrahydrofuran in
polar aprotic solvents such as tetrahydrofuran, diethyl ether, 1,4-dioxane or
1,2-
dimethoxyethane at temperatures ranging from 0 C to 110 C for 1 to 24 hours.
Typical
conditions include the use of borane-dimethylsulfide in tetrahydrofuran at 70
C for 14
hours.
In order to prepare compounds of Formula F from compounds of Formula E, a
cyano group must be introduced (Step 4) This may be achieved via a range of
conditions. One method of cyano group introduction may be the use of a copper
salt
such as copper cyanide in a polar aprotic solvent such as N,N-
dimethylformamide
(DMF), N-methylpyrrolidinone (NMP), N,N-dimethylacetamide (DMA) at
temperatures
ranging from 22 C to 200 C for 1 to 24 hours. Copper cyanide in N,N-
dimethylformamide heated at 165 C for 5 hours is a typical protocol for this
transformation.
Alternatively in Step 4, alkali cyanide salts such as potassium or sodium
cyanide
may be used in conjunction with catalysts such as 18-crown-6 (U52005020564)
and or
tetrabutylammonium bromide (J. Med. Chem. 2003, 46, 1144) in polar aprotic
solvents
such acetonitrile and dimethylsulfoxide at temperatures ranging from 22 C to
100 C for
the addition of a cyano group to this template.
Finally, the use of metal catalysis is common for the transformation depicted
in
Step 4. Common cyanide salts used in catalytic procedures include zinc
cyanide,
copper cyanide, sodium cyanide, and potassium hexacyanoferrate (II). The metal
catalysts can be copper catalysts such as copper iodide and or palladium
catalysts such
as tris(dibenzylideneacetone)dipalladium (Pd2(dba)3), palladium tetrakis-
triphenylphosphine (Pd(PPh3)4), or dichloro(diphenyl-phosphinoferrocene)-
palladium
(Pd(dppf)Cl2). These catalysts may be used alone or in any combination with
any of the
above cyanide salts. To these reactions may be added ligands such as 1,1'-
bis(diphenylphosphino)-ferrocene (dppf) or metal additives such as zinc or
copper metal.
The reactions are carried out in polar aprotic solvents such as NMP, DMF, DMA
with or
without water as an additive. The reactions are carried out at temperatures
ranging from
22 C to 150 C via conventional or microwave heating for 1 to 48 hours and may
be
conducted in a sealed or non-sealed reaction vessel. Typical conditions for
Step 4
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include the use of zinc cyanide, Pd2(dba)3, dppf, and zinc dust in DMA heated
at 120 C
in a microwave for 1 hour (J. Med. Chem. 2005, 48, 1132).
In Step 5, compounds of Formula G, can be synthesized from compounds of
Formula F via the Mitsunobu reaction. The Mitusunobu reaction has been
reviewed in
the synthetic literature (e.g., Chem. Asian. J. 2007, 2, 1340; Eur. J. Org.
Chem. 2004,
2763; S. Chem. Eur. J. 2004, 10, 3130), and many of the synthetic protocols
listed in
these reviews may be used. The use of Mitsunobu reaction protocols utilizing
azodicarboxylates such as diethyl azodicarboxylate (DEAD), di-tert-butyl
azodicarboxylate (TBAD), diisopropyl azodicarboxylate (DIAD) and a phosphine
reagent
such as triphenylphosphine (PPh3), tributylphoshine (PBu3) and polymer
supported
triphenylphosphine (PS-PPh3) are combined with compounds of Formula F and a
compound of general structure X-OH. Solvents utilized in this reaction may
include
aprotic solvents such as toluene, benzene, THF, 1,4-dioxane and acetonitrile
at
temperatures ranging from 0 C to 130 C depending on the solvent and
azodicarboxylates utilized. Typical conditions for this transformation are the
use of
DEAD with PS-PPh3 in 1,4-dioxane at 22 C for 15 hours.
An alternative to the Mitsunobu reaction for preparing compounds of Formula G
is to convert the compounds of Formula F to the corresponding methanesulfonate
or
para-toluenesulphonate derivatives using methanesulfonyl chloride or para-
toluenesulfonyl chloride, respectively, in the presence of a base such as
triethylamine or
pyridine. The intermediate sulfonate ester is then combined with a compound of
general
X-OH, in the presence of a base such as potassium carbonate, sodium hydride,
or
potassium tert-butoxide to yield compounds of Formula G.
Compounds of Formula K, wherein R1 is C1-C4 alkyl or C3-C6 cycloalkyl, may be
prepared from compounds of Formula F in three Steps: 1) oxidation of the
primary
alcohol to the corresponding aldehyde of Formula H (Step 6, Scheme 1), 2)
reaction of
the aldehyde intermediate of Formula H with an organometallic reagent of the
Formula
R1M, wherein M is lithium (Li) or magnesium halide (MgCI, MgBr or Mg1) to
provide a
secondary alcohol of Formula J, wherein R1 is C1-C4 alkyl or C3-C6 cycloalkyl
(Step 7),
and 3) reaction of the secondary alcohol of Formula J with a phenol of the
Formula X-
OH under Mitsunobu reaction conditions (Step 8).
In Step 6 (Scheme 1), compounds of Formula H can are formed via oxidation
procedures including the use of 1 to 20 equivalents of activated manganese
dioxide in
solvents including but not limited to dichloromethane, acetonitrile, hexane or
acetone
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alone or in combinations for 1 to 72 hours at 22 C to 80 C. Alternatively,
this oxidation
can be conducted with 1 to 3 equivalents of trichloroisocyanuric acid in the
presence of
0.1 to 1 equivalents of 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) in
dichloromethane
or chloroform at temperatures ranging from 0 C to 22 C for 0.1 to 12 hours.
Typical
conditions for this transformation are the use of trichloroisocyanuric acid in
the presence
of 0.1 equivalent of TEMPO in dichloromethane at 22 C for 1 hour.
The preparation of compounds of this invention wherein Y is NR5 is also shown
in Scheme 1. Compounds of Formula L may be prepared from the intermediate
compound of Formula H (Scheme 1) by reaction with an amino compound of the
Formula X-NH-R5 under reductive amination conditions (Step 9) (J. Org. Chem.,
1996,
61, 3849; Org. React. 2002, 59, 1). Similarly compounds of Formula N, wherein
R1 is
Crat alkyl or C3-C6 cycloalkyl may be prepared in two steps from the
intermediate of
Formula J wherein R1 is C1-C4 alkyl or C3-C6 cycloalkyl, by 1) oxidation to
the
corresponding ketone of Formula M (Step 10), and 2) reaction of the ketone of
Formula
M with an amino compound of the Formula X-NH-R5 under reductive amination
conditions (Step 11). Alternatively compounds of Formula L and Formula N,
wherein R5
is C1-C4 alkyl may be prepared from the corresponding compounds of Formula L,
wherein R5 is H, or the corresponding compounds of Formula N, wherein R5 is H,
by
alkylation with an alkyl halide of Formula (C1-C4)-CI, (Ci-C4)-Br or (C1-C4)-I
in the
presence of a base.
Compounds of this invention may also be prepared as shown in Schemes 2 and
3, wherein X, Z, R1, R2a, R3, R4, R5, R6, R7, .-s8
K and R9 are the same as described in
Scheme 1. In particular, compounds of Formula R may be prepared as shown in
Scheme 2.
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Scheme 2
H
OHC
) \\ Step 1 / \\
NC N-N
__________________________________________ NC ,NN
R2.9,,,...),,,,,
R2....õ,,,,,L,
...===
N ...--
Z N
H Z 0
Step 2 X¨P
V
x X
Step 3
-4
i / __ \\
N
NC N-N NC N,
R20,.,.. R2a....õ,...c
....,
/ N
N
R Z
Z Q
In Step 1 of Scheme 2, compounds of the Formula 0 can be formed from
aldehydes of Formula H (see also Scheme 1) via the use of either dimethyl
(diazomethyl)phosphonate or dimethy1-1-diazo-2-oxopropylphosphonate and bases
such as potassium carbonate or potassium tert-butoxide in solvents including
methanol,
ethanol or tetrahydrofuran at temperatures ranging from -78 C to 22 C for 0.1
to 24
hours. Typical conditions for this transformation include the use of dimethy1-
1-diazo-2-
oxopropylphosphonate and 2 equivalents of potassium carbonate in methanol at
22 C
for 0.75 hour.
In Step 2, compounds of Formula Q can be formed from compounds of Formula
0 via a metal-catalyzed Sonagashira coupling procedure with compounds of
general
structure X-P wherein P is a halide or trifluoromethsulfonate (triflate). The
Sonogashira
reaction has been extensively reviewed (Chem. Rev. 2007, 107, 874; Angew.
Chem. Int.
Ed. 2007, 46, 834; Angew. Chem. Int. Ed. 2008, 47, 6954), and many of the
synthetic
protocols listed in these reviews may be used for the synthesis of compounds
of
Formula Q. Typically, the use of metal catalysts in this reaction can be
copper catalysts
such as copper iodide and or palladium catalysts such as Pd2(dba)3, Pd(PPh3)4,
22
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Pd(dppf)Cl2 or Pd(PPh3)2Cl2. These catalysts may be used alone or in any
combination.
Base additives are typically used in this reaction and may include amine bases
such as
diethylamine, triethylamine, diisopropylethylamine or pyrrolidine or inorganic
bases such
as potassium carbonate or potassium fluoride. The reactions are carried out in
solvents
such as dichloromethane, chloroform, acetonitrile, DMF, toluene or 1,4-dioxane
with or
without water as an additive. The reactions are carried out at temperatures
ranging from
0 C to 150 C depending on the solvent for times ranging from 0.1 to 48 hours.
Typical
conditions for this transformation include the use of Cul and Pd(PPh3)2Cl2 in
DMF at
90 C for 2 hours.
Finally, in Step 3 compounds of Formula R can be formed from compounds of
Formula Q via hydrogenation in the presence of transition metal catalysts.
Common
catalysts include the use of 5 ¨ 20% palladium on carbon or 5 ¨ 20% palladium
hydroxide on carbon. These reactions can be conducted in a Parr shaker
apparatus or
in an H-Cube hydrogenation flow reactor (ThalesNano, U.K.) under pressures of
hydrogen ranging from 1 to 50 psi in polar solvents such as tetrahydrofuran,
ethyl
acetate, methanol or ethanol at temperatures of 22 C to 50 C for times ranging
from 0.1
to 24 hours. Typical conditions for Step 3 include the use compound of Formula
Q in
ethyl acetate at a flow rate of 1 mL/min through a 10% palladium on carbon
cartridge on
the H-Cube flow apparatus set at the "full hydrogen" setting.
Scheme 3 shows methods for the preparation of compounds of Formula W.
23
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Scheme 3
OH OHC e e x
)
1R5
Br PPh3 \\
,\\N NC N X¨( -N
NC N ___________________________________________ ...-
R2...,.._õõ--1.,, Step 5 NC N-
R2zõ..),...õ.
N
.-..
N 1
...--
1 F Z H N
Z
z1 v
Step 1 Step 3 0
X4 Step 4
,
R5
U
Br e (D x
--R5
Br PPh3
___ \\m Step 2
NC N-- ..
\\ ) \\
R2 NC N-N NC N-N
,?......c1,,,, R2,.,....õ..,..-1.,, R2,..,
N
Z
S N T N W i
Z Z
In Step 1 of Scheme 3, compounds of Formula F (see also Scheme 2) can be
treated with reagents such as phosphorus tribromide or carbon tetrabromide and
triphenylphosphine to give compounds of Formula S. In Step 2, compounds of
Formula
S are then allowed to react with triphenylphosphine in solvents such as
dichloromethane, chloroform, toluene, benzene, tetrahydrofu ran (THF) or
acetonitrile to
give triphenylphosphonium salts of Formula T. The salts of Formula T, are then
combined with carbonyl compounds of Formula U in the presence of bases such as
n-
butyllithium, sodium bis(trimethylsilyl)amide, lithium
bis(trimethylsilyl)amide, potassium
bis(trimethylsilyl)amide or lithium diisopropylamide in solvents such as THF,
diethylether
or 1,4-dioxane, to yield alkene compounds of Formula V, which are typically
isolated as
mixtures of E and Z geometric isomers (Step 3). This reaction, commonly known
as the
Wittig olefination reaction, has been reviewed extensively in the literature
(Chem. Rev.
1989, 89, 863; Modern Carbonyl Olefination 2004, 1-17; Liebigs Ann.Chem. 1997,
1283).
In Step 4, compounds of Formula W are formed from compounds of Formula V
via hydrogenation in the presence of transition metal catalysts. Common
catalysts
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include the use of 5 ¨ 20% palladium on carbon or 5 ¨ 20% palladium hydroxide
on
carbon. These reactions can be conducted in a similar manner as described for
Step 3
of Scheme 2.
Alternatively compounds of Formula W may be prepared from aldehydes of
Formula H via Wittig reaction with triphenylphosphonium salts of Formula AA
(Step 5,
Scheme 3). As for Step 3, this reaction produces alkene compounds of Formula
V,
which again are typically isolated as mixtures of E and Z geometric isomers,
and may
be converted to compounds of Formula W by hydrogenation. The salts of Formula
AA
are obtained in a similar manner to that used for preparing salts of Formula T
via
conversion of the corresponding alcohol to the bromide and subsequent reaction
with
triphenylphosphine.
Compounds of Formula BB shown below, wherein X, Z, R1 and R2a are as
defined in Scheme 1 can be prepared from secondary alcohols of Formula J (see
Scheme 2) or ketones of Formula M (see Scheme 2) through reaction sequences
similar to those shown in Scheme 3. Conversion of compounds of Formula J to
the
corresponding bromides, followed by Wittig olefination with aldehydes of
general
formula X-CHO provides alkenes of Formula CC. Alkenes of Formula CC may also
be
obtained via Wittig reaction of ketones of Formula M with salts of the general
structure
X-CH2-PPh3+13(. The alkenes of Formula CC are then converted to compounds of
Formula BB by hydrogenation.
x x
R1
R1
NC "N' NC N,
N BB N CC
i 1
Z Z
In certain instances it is possible to change the order of steps shown in
Schemes
1, 2 and 3. For example, in Scheme 1, it is sometimes possible to introduce
the cyano
group on the pyrazole ring as the last step, i.e., inverting the order in
which Steps 4 and
5 are carried out. Also, in certain cases, it is preferable to introduce or
modify
substituents R3 on the group X later in the synthesis, even as the last step.
For example,
when R3 is 502R7, the 502R7 group may be in formed in the last step by
oxidation of
the corresponding compound bearing a substituent of general formula S-R7.
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Compounds of this invention may be prepared according to sequences
analogous to those shown in Schemes 1, 2 and 3 starting with 3,3-difluoro-4,4-
dihydroxy 1-piperidine carboxylic acid 1,1-dimethylethyl ester (WO
2008121687). In a
manner similar to that described for the preparation of intermediates of
formula A in
Scheme 1, this material may be converted to hydrazine dervatives of formula
DD, which
are then used similarly to the intermediates of formula A in Scheme 1.
,NH2
HN
F>
F
N DD
Z
As is readily apparent to one skilled in the art, protection of remote
functionality
(e.g., primary or secondary amine) of intermediates may be necessary. The need
for
such protection will vary depending on the nature of the remote functionality
and the
conditions of the preparation methods. Suitable amino-protecting groups (NH-
Pg)
include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl
(CBZ) and 9-
fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a "hydroxy-protecting group"
refers to
a substituent of a hydroxy group that blocks or protects the hydroxy
functionality.
Suitable hydroxyl-protecting groups (0-Pg) include for example, allyl, acetyl,
silyl,
benzyl, para-methoxybenzyl, trityl, and the like. The need for such protection
is readily
determined by one skilled in the art. For a general description of protecting
groups and
their use, see T. W. Greene, Protective Groups in Organic Synthesis, John
Wiley &
Sons, New York, 1991.
As noted above, some of the compounds of this invention are acidic and they
form salts with pharmaceutically acceptable cations. Some of the compounds of
this
invention are basic and form salts with pharmaceutically acceptable anions.
All such
salts are within the scope of this invention and they can be prepared by
conventional
methods such as combining the acidic and basic entities, usually in a
stoichiometric
ratio, in either an aqueous, non-aqueous or partially aqueous medium, as
appropriate.
The salts are recovered either by filtration, by precipitation with a non-
solvent followed
by filtration, by evaporation of the solvent, or, in the case of aqueous
solutions, by
lyophilization, as appropriate. The compounds are obtained in crystalline form
according
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to procedures known in the art, such as by dissolution in an appropriate
solvent(s) such
as ethanol, hexanes or water/ethanol mixtures.
Medical Uses
Compounds of the present invention modulate the activity of G-protein-coupled
receptor GPR119. As such, said compounds are useful for the prophylaxis and
treatment of diseases, such as diabetes, in which the activity of GPR119
contributes to
the pathology or symptoms of the disease. Consequently, another aspect of the
present invention includes a method for the treatment of a metabolic disease
and/or a
metabolic-related disorder in an individual which comprises administering to
the
individual in need of such treatment a therapeutically effective amount of a
compound of
the invention, a salt of said compound or a pharmaceutical composition
containing such
compound. The metabolic diseases and metabolism-related disorders are selected
from,
but not limited to, hyperlipidemia, Type I diabetes, Type II diabetes
mellitus, idiopathic
Type I diabetes (Type lb), latent autoimmune diabetes in adults (LADA), early-
onset
Type 2 diabetes (EOD), youth-onset atypical diabetes (YOAD), maturity onset
diabetes
of the young (MODY), malnutrition-related diabetes, gestational diabetes,
coronary
heart disease, ischemic stroke, restenosis after angioplasty, peripheral
vascular disease,
intermittent claudication, myocardial infarction (e.g. necrosis and
apoptosis),
dyslipidemia, post-prandial lipemia, conditions of impaired glucose tolerance
(IGT),
conditions of impaired fasting plasma glucose, metabolic acidosis, ketosis,
arthritis,
obesity, osteoporosis, hypertension, congestive heart failure, left
ventricular hypertrophy,
peripheral arterial disease, diabetic retinopathy, macular degeneration,
cataract,
diabetic nephropathy, glomerulosclerosis, chronic renal failure, diabetic
neuropathy,
metabolic syndrome, syndrome X, premenstrual syndrome, coronary heart disease,
angina pectoris, thrombosis, atherosclerosis, myocardial infarction, transient
ischemic
attacks, stroke, vascular restenosis, hyperglycemia, hyperinsulinemia,
hyperlipidemia,
hypertrygliceridemia, insulin resistance, impaired glucose metabolism,
conditions of
impaired glucose tolerance, conditions of impaired fasting plasma glucose,
obesity,
erectile dysfunction, skin and connective tissue disorders, foot ulcerations,
endothelial
dysfunction, hyper apo B lipoproteinemia and impaired vascular compliance.
Additionally, the compounds may be used to treat neurological disorders such
as
Alzheimer's disease, schizophrenia, and impaired cognition. The compounds will
also
be beneficial in gastrointestinal illnesses such as inflammatory bowel
disease, ulcerative
colitis, Crohn's disease, irritable bowel syndrome, etc. As noted above the
compounds
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may also be used to stimulate weight loss in obese patients, especially those
afflicted
with diabetes.
In accordance with the foregoing, the present invention further provides a
method
for preventing or ameliorating the symptoms of any of the diseases or
disorders
described above in a subject in need thereof, which method comprises
administering to
a subject a therapeutically effective amount of a compound of the present
invention.
Further aspects of the invention include the preparation of medicaments for
the treating
diabetes and its related co-morbidities.
In order to exhibit the therapeutic properties described above, the compounds
need to be administered in a quantity sufficient to modulate activation of the
G-protein-
coupled receptor GPR119. This amount can vary depending upon the particular
disease/condition being treated, the severity of the patient's
disease/condition, the
patient, the particular compound being administered, the route of
administration, and
the presence of other underlying disease states within the patient, etc. When
administered systemically, the compounds typically exhibit their effect at a
dosage
range of from about 0.1 mg/kg/day to about 100 mg/kg/day for any of the
diseases or
conditions listed above. Repetitive daily administration may be desirable and
will vary
according to the conditions outlined above.
The compounds of the present invention may be administered by a variety of
routes. They may be administered orally. The compounds may also be
administered
parenterally (i.e., subcutaneously, intravenously, intramuscularly,
intraperitoneally, or
intrathecally), rectally, or topically.
Co-Administration
The compounds of this invention may also be used in conjunction with other
pharmaceutical agents for the treatment of the diseases, conditions and/or
disorders
described herein. Therefore, methods of treatment that include administering
compounds of the present invention in combination with other pharmaceutical
agents
are also provided. Suitable pharmaceutical agents that may be used in
combination
with the compounds of the present invention include anti-obesity agents
(including
appetite suppressants), anti-diabetic agents, anti-hyperglycemic agents, lipid
lowering
agents, and anti-hypertensive agents.
Suitable anti-diabetic agents include an acetyl-CoA carboxylase-2 (ACC-2)
inhibitor, a diacylglycerol 0-acyltransferase 1 (DGAT-1) inhibitor, a
phosphodiesterase
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(PDE)-10 inhibitor, a sulfonylurea (e.g., acetohexamide, chlorpropamide,
diabinese,
glibenclamide, glipizide, glyburide, glimepiride, gliclazide, glipentide,
gliquidone,
glisolamide, tolazamide, and tolbutamide), a meglitinide, an a-amylase
inhibitor (e.g.,
tendamistat, trestatin and AL-3688), an a-glucoside hydrolase inhibitor (e.g.,
acarbose),
an a-glucosidase inhibitor (e.g., adiposine, camiglibose, emiglitate,
miglitol, voglibose,
pradimicin-Q, and salbostatin), a PPARy agonist (e.g., balaglitazone,
ciglitazone,
darglitazone, englitazone, isaglitazone, pioglitazone, rosiglitazone and
troglitazone), a
PPAR a/y agonist (e.g., CLX-0940, GW-1536, GW-1929, GW-2433, KRP-297, L-
796449, LR-90, MK-0767 and SB-219994), a biguanide (e.g., metformin), a
glucagon-
like peptide 1 (GLP-1) agonist (e.g., exendin-3 and exendin-4), a protein
tyrosine
phosphatase-1B (PTP-1B) inhibitor (e.g., trodusquemine, hyrtiosal extract, and
compounds disclosed by Zhang, S., et al., Drug Discovery Today, 12(9/10), 373-
381
(2007)), SIRT-1 inhibitor (e.g., reservatrol), a dipeptidyl peptidease IV (DPP-
IV) inhibitor
(e.g., sitagliptin, vildagliptin, alogliptin and saxagliptin), an insulin
secreatagogue, a fatty
acid oxidation inhibitor, an A2 antagonist, a c-jun amino-terminal kinase
(JNK) inhibitor,
insulin, an insulin mimetic, a glycogen phosphorylase inhibitor, a VPAC2
receptor
agonist, and a SGLT2 inhibitor (sodium dependent glucose transporter
inhibitors such
as dapagliflozin, etc). Preferred anti-diabetic agents are metformin and DPP-
IV
inhibitors (e.g., sitagliptin, vildagliptin, alogliptin and saxagliptin).
Suitable anti-obesity agents include 1113-hydroxy steroid dehydrogenase-1
(1113-
HSD type 1) inhibitors, stearoyl-CoA desaturase-1 (SCD-1) inhibitor, MCR-4
agonists,
cholecystokinin-A (CCK-A) agonists, monoamine reuptake inhibitors (such as
sibutramine), sympathomimetic agents, 133 adrenergic agonists, dopamine
agonists
(such as bromocriptine), melanocyte-stimulating hormone analogs, 5HT2c
agonists,
melanin concentrating hormone antagonists, leptin (the OB protein), leptin
analogs,
leptin agonists, galanin antagonists, lipase inhibitors (such as
tetrahydrolipstatin, i.e.
orlistat), anorectic agents (such as a bombesin agonist), neuropeptide-Y
antagonists
(e.g., NPY Y5 antagonists), PYY3_36(including analogs thereof), thyromimetic
agents,
dehydroepiandrosterone or an analog thereof, glucocorticoid agonists or
antagonists,
orexin antagonists, glucagon-like peptide-1 agonists, ciliary neurotrophic
factors (such
as AxokineTM available from Regeneron Pharmaceuticals, Inc., Tarrytown, NY and
Procter & Gamble Company, Cincinnati, OH), human agouti-related protein (AGRP)
inhibitors, ghrelin antagonists, histamine 3 antagonists or inverse agonists,
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neuromedin U agonists, MTP/ApoB inhibitors (e.g., gut-selective MTP
inhibitors, such
as dirlotapide), opioid antagonist, orexin antagonist, and the like.
Preferred anti-obesity agents for use in the combination aspects of the
present
invention include gut-selective MTP inhibitors (e.g., dirlotapide, mitratapide
and
implitapide, R56918 (CAS No. 403987) and CAS No. 913541-47-6), CCKa agonists
(e.g., N-benzy1-244-(1H-indo1-3-ylmethyl)-5-oxo-1-phenyl-4,5-dihydro-2,3,6,10b-
tetraaza-benzo[e]azulen-6-y1]-N-isopropyl-acetamide described in PCT
Publication No.
WO 2005/116034 or US Publication No. 2005-0267100 Al), 5HT2c agonists (e.g.,
lorcaserin), MCR4 agonist (e.g., compounds described in US 6,818,658), lipase
inhibitor (e.g., Cetilistat), PYY3_36(as used herein "PYY3_36" includes
analogs, such as
peglated PYY3_36 e.g., those described in US Publication 2006/0178501), opioid
antagonists (e.g., naltrexone), oleoyl-estrone (CAS No. 180003-17-2),
obinepitide
(TM30338), pramlintide (Symlin ), tesofensine (N52330), leptin, liraglutide,
bromocriptine, orlistat, exenatide (Byetta ), AOD-9604 (CAS No. 221231-10-3)
and
sibutramine. Preferably, compounds of the present invention and combination
therapies
are administered in conjunction with exercise and a sensible diet.
All of the above recited U.S. patents and publications are incorporated herein
by
reference.
Pharmaceutical Formulations
The present invention also provides pharmaceutical compositions which
comprise a therapeutically effective amount of a compound, or a
pharmaceutically
acceptable salt thereof, in admixture with at least one pharmaceutically
acceptable
excipient. The compositions include those in a form adapted for oral, topical
or
parenteral use and can be used for the treatment of diabetes and related
conditions as
described above.
The composition can be formulated for administration by any route known in the
art, such as subdermal, inhalation, oral, topical, parenteral, etc. The
compositions may
be in any form known in the art, including but not limited to tablets,
capsules, powders,
granules, lozenges, or liquid preparations, such as oral or sterile parenteral
solutions or
suspensions.
Tablets and capsules for oral administration may be in unit dose presentation
form, and may contain conventional excipients such as binding agents, for
example
syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrollidone;
fillers, for example
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lactose, sugar, maize-starch, calcium phosphate, sorbitol or glycine;
tabletting lubricants,
for example magnesium stearate, talc, polyethylene glycol or silica;
disintegrants, for
example potato starch; or acceptable wetting agents such as sodium lauryl
sulphate.
The tablets may be coated according to methods well known in normal
pharmaceutical
practice.
Oral liquid preparations may be in the form of, for example, aqueous or oily
suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a
dry
product for reconstitution with water or other suitable vehicle before use.
Such liquid
preparations may contain conventional additives, such as suspending agents,
for
example sorbitol, methyl cellulose, glucose syrup, gelatin, hydroxyethyl
cellulose,
carboxymethyl cellulose, aluminium stearate gel or hydrogenated edible fats,
emulsifying agents, for example lecithin, sorbitan monooleate, or acacia; non-
aqueous
vehicles (which may include edible oils), for example almond oil, oily esters
such as
glycerin, propylene glycol, or ethyl alcohol; preservatives, for example
methyl or propyl
p-hydroxybenzoate or sorbic acid, and, if desired, conventional flavoring or
coloring
agents.
For parenteral administration, fluid unit dosage forms are prepared utilizing
the
compound and a sterile vehicle, water being preferred. The compound, depending
on
the vehicle and concentration used, can be either suspended or dissolved in
the vehicle
or other suitable solvent. In preparing solutions, the compound can be
dissolved in
water for injection and filter sterilized before filling into a suitable vial
or ampoule and
sealing. Advantageously, agents such as local anesthetics, preservatives and
buffering
agents etc. can be dissolved in the vehicle. To enhance the stability, the
composition
can be frozen after filling into the vial and the water removed under vacuum.
The dry
lyophilized powder is then sealed in the vial and an accompanying vial of
water for
injection may be supplied to reconstitute the liquid prior to use. Parenteral
suspensions
are prepared in substantially the same manner except that the compound is
suspended
in the vehicle instead of being dissolved and sterilization cannot be
accomplished by
filtration. The compound can be sterilized by exposure to ethylene oxide
before
suspending in the sterile vehicle. Advantageously, a surfactant or wetting
agent is
included in the composition to facilitate uniform distribution of the
compound.
The compositions may contain, for example, from about 0.1% to about 99 by
weight, of the active material, depending on the method of administration.
Where the
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compositions comprise dosage units, each unit will contain, for example, from
about 0.1
to 900 mg of the active ingredient, more typically from 1 mg to 250mg.
Compounds of the invention can be formulated for administration in any
convenient way for use in human or veterinary medicine, by analogy with other
anti-
diabetic agents. Such methods are known in the art and have been summarized
above.
For a more detailed discussion regarding the preparation of such formulations;
the
reader's attention is directed to Remington"s Pharmaceutical Sciences, 21st
Edition, by
University of the Sciences in Philadelphia.
Embodiments of the present invention are illustrated by the following
Examples.
It is to be understood, however, that the embodiments of the invention are not
limited to
the specific details of these Examples, as other variations thereof will be
known, or
apparent in light of the instant disclosure, to one of ordinary skill in the
art.
EXAMPLES
Unless specified otherwise, starting materials are generally available from
commercial sources such as Aldrich Chemicals Co. (Milwaukee, WI), Lancaster
Synthesis, Inc. (Windham, NH), Acros Organics (Fairlawn, NJ), Maybridge
Chemical
Company, Ltd. (Cornwall, England), Tyger Scientific (Princeton, NJ), and
AstraZeneca
Pharmaceuticals (London, England), Mallinckrodt Baker (Phillipsburg NJ); EMD
(Gibbstown, NJ).
General Experimental Procedures
NMR spectra were recorded on a Varian UnityTM 400 (DG400-5 probe) or 500
(DG500-5 probe ¨ both available from Varian Inc., Palo Alto, CA) at room
temperature
at 400 MHz or 500 MHz respectively for proton analysis. Chemical shifts are
expressed
in parts per million (delta) relative to residual solvent as an internal
reference. The peak
shapes are denoted as follows: s, singlet; d, doublet; dd, doublet of doublet;
t, triplet; q,
quartet; m, multiplet; bs, broad singlet; 2s, two singlets.
Atmospheric pressure chemical ionization mass spectra (APCI) were obtained on
a Waters TM Spectrometer (Micromass ZMD, carrier gas: nitrogen) (available
from
Waters Corp., Milford, MA, USA) with a flow rate of 0.3 mL/minute and
utilizing a 50:50
water/acetonitrile eluent system. Electrospray ionization mass spectra (ES)
were
obtained on a liquid chromatography mass spectrometer from Waters TM
(Micromass ZQ
or ZMD instrument (carrier gas: nitrogen) (Waters Corp., Milford, MA, USA)
utilizing a
gradient of 95:5 ¨ 0:100 water in acetonitrile with 0.01% formic acid added to
each
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solvent. These instruments utilized a Varian Polaris 5 C18-A20x2.0mm column
(Varian
Inc., Palo Alto, CA) at flow rates of 1mL/minute for 3.75 minutes or 2
mL/minute for 1.95
minutes.
Column chromatography was performed using silica gel with either Flash 40
BiotageTm columns (ISC, Inc., Shelton, CT) or BiotageTM SNAP cartridge KPsil
or
Redisep Rf silica (from Teledyne Isco Inc) under nitrogen pressure.
Preparative HPLC
was performed using a Waters FractionLynx system with photodiode array (Waters
2996) and mass spectrometer (Waters/Micromass ZQ) detection schemes.
Analytical
HPLC work was conducted with a Waters 2795 Alliance HPLC or a Waters ACQUITY
UPLC with photodiode array, single quadrupole mass and evaporative light
scattering
detection schemes.
Concentration in vacuo refers to evaporation of solvent under reduced pressure
using a rotary evaporator.
Unless otherwise noted, chemical reactions were performed at room temperature
(about 23 degrees Celsius). Also, unless otherwise noted chemical reactions
were run
under an atmosphere of nitrogen.
PHARMACOLOGICAL DATA
The practice of the invention for the treatment of diseases modulated by the
agonist activation of G-protein-coupled receptor GPR119 with compounds of the
invention can be evidenced by activity in one or more of the functional assays
described
herein below. The source of supply is provided in parenthesis.
33
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In-Vitro Functional Assays
R-lactamase:
The assay for GPR119 agonists utilizes a cell-based (hGPR119 HEK293-CRE
beta-lactamase) reporter construct where agonist activation of human GPR119 is
coupled to beta-lactamase production via a cyclic AMP response element
(CRE). GPR119 activity is then measured utilizing a FRET-enabled beta-
lactamase
substrate, CCF4-AM (Live Blazer FRET-B/G Loading kit, lnvitrogen cat #
K1027). Specifically, hGPR119-HEK-CRE- beta-lactamase cells (Invitrogen 2.5 x
107/mL) were removed from liquid nitrogen storage, and diluted in plating
medium
(Dulbecco's modified Eagle medium high glucose (DMEM; Gibco Cat # 11995-065),
10% heat inactivated fetal bovine serum (HIFBS; Sigma Cat # F4135), 1X MEM
Nonessential amino acids (Gibco Cat # 15630-080), 25 mM HEPES pH 7.0 (Gibco
Cat
#15630-080), 200 nM potassium clavulanate (Sigma Cat # P3494). The cell
concentration was adjusted using cell plating medium and 50 microL of this
cell
suspension (12.5 x 10 viablecells) was added into each well of a black, clear
bottom,
poly-d-lysine coated 384-well plate (Greiner Bio-One cat# 781946) and
incubated at 37
degrees Celsius in a humidified environment containing 5% carbon dioxide.
After 4
hours the plating medium was removed and replaced with 40 microL of assay
medium
(Assay medium is plating medium without potassium clavulanate and HIFBS).
Varying
concentrations of each compound to be tested was then added in a volume of 10
microL (final DMSO 0.5%) and the cells were incubated for 16 hours at 37
degrees
Celsius in a humidified environment containing 5% carbon dioxide. Plates were
removed from the incubator and allowed to equilibrate to room temperature for
approximately 15 minutes. 10 microL of 6 X CCF4/AM working dye solution
(prepared
according to instructions in the Live Blazer FRET-B/G Loading kit, lnvitrogen
cat #
K1027) was added per well and incubated at room temperature for 2 hours in the
dark.
Fluorescence was measured on an EnVision fluorimetric plate reader, excitation
405
nm, emission 460 nm/535 nm. EC50 determinations were made from agonist-
response
curves analyzed with a curve fitting program using a 4-parameter logistic dose-
response
equation.
cAMP:
GPR119 agonist activity was also determined with a cell-based assay utilizing
an
HTRF (Homogeneous Time-Resolved Fluorescence) cAMP detection kit (cAMP
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dynamic 2 Assay Kit; Cis Bio cat # 62AM4PEC) that measures cAMP levels in the
cell.
The method is a competitive immunoassay between native cAMP produced by the
cells
and the cAMP labeled with the dye d2. The tracer binding is visualized by a
Mab ant
cAMP labeled with Cryptate. The specific signal (Le. energy transfer) is
inversely
proportional to the concentration of cAMP in either standard or sample.
Specifically. hGPR119 HEK-CRE beta-lactamase cells (I nvitrogen 2.5 x 107/mL;
the same cell line used in the beta-lactamase assay described above) were
removed
from cryopreservation and diluted in growth medium (Dulbecco's modified Eagle
medium high glucose (DMEM; Gibco Cat # 11995-065), 1% charcoal dextran treated
fetal bovine serum (CD serum; HyClone Cat # SH30068.03), lx MEM Nonessential
amino acids (Gibco Cat # 15630-080) and 25 mM HEPES pH 7.0 (Gibco Cat # 15630-
080)). The cell concentration was adjusted to 1.5 x 105 cells/mL and 30 mLs of
this
suspension was added to a T-175 flask and incubated at 37 degrees Celsius in a
humidified environment in 5% carbon dioxide. After 16 hours (overnight), the
cells were
removed from the T-175 flask (by rapping the side of the flask), centrifuged
at 800 x g
and then re-suspended in assay medium (lx HBSS +CaCl2+ MgC12 (Gibco Cat #
14025-092) and 25 mM HEPES pH 7.0 (Gibco Cat # 15630-080)). The cell
concentration was adjusted to 6.25 x 105 cells/mL with assay medium and 8 pl
of this
cell suspension (5000 cells) was added to each well of a white Greiner 384-
well, low-
volume assay plate (VWR cat # 82051-458).
Varying concentrations of each compound to be tested were diluted in assay
buffer containing 3-isobuty1-1-methylxanthin (IBMX; Sigma cat #I5879) and
added to
the assay plate wells in a volume of 2 microL (final IBMX concentration was
400 microM
and final DMSO concentration was 0.58%). Following 30 minutes incubation at
room
temperature, 5 microL of labeled d2 cAMP and 5 microL of anti-cAMP antibody
(both
diluted 1:20 in cell lysis buffer; as described in the manufacturers assay
protocol) were
added to each well of the assay plate. The plates were then incubated at room
temperature and after 60 minutes, changes in the HTRF signal were read with an
Envision 2104 multilabel plate reader using excitation of 330 nm and emissions
of 615
and 665 nm. Raw data were converted to nM cAMP by interpolation from a cAMP
standard curve (as described in the manufacturer's assay protocol) and EC50
determinations were made from an agonist-response curves analyzed with a curve
fitting program using a 4-paramter logistic dose response equation.
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It is recognized that cAMP responses due to activation of GPR119 could be
generated in cells other than the specific cell line used herein.
R-Arrestin:
GPR119 agonist activity was also determined with a cell-based assay utilizing
DiscoverX PathHunter R-arrestin cell assay technology and their U205 hGPR119
R-arrestin cell line (DiscoverX Cat # 93-0356C3). In this assay, agonist
activation is
determined by measuring agonist-induced interaction of 13-arrestin with
activated
GPR119. A small, 42 amino acid enzyme fragment, called ProLink was appended to
the
Specifically, U205 hGPR119 R-arrestin cells (DiscoverX 1 x 107/mL) were
removed from cryopreservation and diluted in growth medium (Minimum essential
medium (MEM; Gibco Cat # 11095-080), 10% heat inactivated fetal bovine serum
(HIFBS; Sigma Cat # F4135-100), 100 mM sodium pyruvate (Sigma Cat # S8636),
500
microg/mL G418 (Sigma Cat # G8168) and 250 microg/mL Hygromycin B (Invitrogen
After 16 hours (overnight) the assay plates were removed from the incubator
and
varying concentrations of each compound to be tested (diluted in assay buffer
(lx
HBSS +CaCl2+ MgC12 (Gibco Cat # 14025-092), 20 mM HEPES pH 7.0 (Gibco Cat #
15630-080) and 0.1% BSA (Sigma Cat # A9576)) were added to the assay plate
wells
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in a volume of 2.5 microL (final DM30 concentration was 0.5 %). After a 90
minute
incubation at 37 degrees Celsius in a humidified environment in 5% carbon
dioxide, 7.5
microL of Galacton Star 8-galactosidase substrate (PathHunter Detection Kit
(DiscoveRx Cat # 93-0001); prepared as described in the manufacturers assay
protocol) was added to each well of the assay plate. The plates were incubated
at room
temperature and after 60 minutes, changes in the luminescence were read with
an
Envision 2104 multilabel plate reader at 0.1 seconds per well. EC50
determinations
were made from an agonist-response curves analyzed with a curve fitting
program
using a 4-parameter logistic dose response equation.
Expression of GPR119 Using BacMam and GPR119 Binding Assay
Wild-type human GPR119 (published in PCT patent publication no.
2010/106457) was amplified via polymerase chain reaction (PCR) (Pfu Turbo
Mater Mix,
Stratagene, La Jolla, CA) using pIRES-puro-hGPR119 as a template and the
following
primers:
hGPR119 BamH1, Upper
5'-TAAATTGGATCCACCATGGAATCATCTTTCTCATTTGGAG-3'
(inserts a BamHI site at the 5' end)
hGPR119 EcoRI, Lower
5'-TAAATTGAATTCTTATCAGCCATCAAACTCTGAGC-3'
(inserts a EcoRI site at the 3' end)
The amplified product was purified (Qiaquick Kit, Qiagen, Valencia, CA) and
digested with BamH1 and EcoRI (New England BioLabs, Ipswich, MA) according to
the
manufacturer's protocols. The vector pFB-VSVG-CMV-poly (published in PCT
patent
publication no. 2010/106457) was digested with BamHI and EcoRI (New England
BioLabs, Ipswich, MA). The digested DNA was separated by electrophoresis on a
1%
agarose gel; the fragments were excised from the gel and purified (Qiaquick
Kit, Qiagen,
Valencia, CA). The vector and gene fragments were ligated (Rapid Ligase Kit,
Roche,
Pleasanton, CA) and transformed into OneShot DH5alpha T1R cells (Invitrogen,
Carlsbad, CA). Eight ampicillin-resistant colonies ("clones 1-8") were grown
for
miniprep (Qiagen Miniprep Kit, Qiagen, Valencia, CA) and sequenced to confirm
identity
and correct insert orientation.
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The pFB-VSVG-CMV-poly-hGPR119 construct (clone #1) was transformed into
OneShot DH10Bac cells (Invitrogen, Carlsbad, CA) according to manufacturers'
protocols. Eight positive (i.e. white) colonies were re-streaked to confirm as
"positives"
and subsequently grown for bacmid isolation. The recombinant hGPR119 bacmid
was
isolated via a modified Alkaline Lysis procedure using the buffers from a
Qiagen
Miniprep Kit (Qiagen, Valencia, CA). Briefly, pelleted cells were lysed in
buffer P1,
neutralized in buffer P2, and precipitated with buffer N3. Precipitate was
pelleted via
centrifugation (17,900xg for 10 minutes) and the supernatant was combined with
isopropanol to precipitate the DNA. The DNA was pelleted via centrifugation
(17,900xg
for 30 minutes), washed once with 70% ethanol, and resuspended in 50 1_
buffer EB
(Tris-HCL, pH 8.5). Polymerase chain reaction (PCR) with commercially
available
primers (M13F, M13R, Invitrogen, Carlsbad, CA) was used to confirm the
presence of
the hGPR119 insert in the Bacmid.
Generation of hGPR119 Recombinant Baculovirus
Creation of PO Virus Stock
Suspension adapted Sf9 cells grown in Sf90011 medium (Invitrogen, Carlsbad,
CA) were
transfected with 10 microL hGPR119 bacmid DNA according to the manufacturer's
protocol (Cellfectin, Invitrogen, Carlsbad, CA). After five days of
incubation, the
conditioned medium (i.e. "PO" virus stock) was centrifuged and filtered
through a 0.22
lirn filter (Steriflip, Millipore, Billerica, MA).
Creation of Frozen Virus (BIIC) Stocks
For long term virus storage and generation of working (i.e. "P1") viral
stocks, frozen BIIC
(Baculovirus Infected Insect Cells) stocks were created as follows: suspension
adapted
Sf9 cells were grown in Sf90011 medium (Invitrogen, Carlsbad, CA) and infected
with
hGPR119 PO virus stock. After 24 hours of growth, the infected cells were
gently
centrifuged (approximately 100 x g), resuspended in Freezing Medium (10% DMSO,
1%
Albumin in Sf90011 medium) to a final density of 1 x 107 cells/mL and frozen
according to
standard freezing protocols in 1 mL aliquots.
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Creation of Working ("P1") Virus Stock
Suspension adapted Sf9 cells grown in Sf90011 medium (Invitrogen, Carlsbad,
CA) were
infected with a 1:100 dilution of a thawed hGPR119 BIIC stock and incubated
for
several days (27 degrees Celsius with shaking). When the viability of the
cells reached
70%, the conditioned medium was harvested by centrifugation and the virus
titer
determined by ELISA (BaculoElisa Kit, Clontech, Mountain View, CA)
Over-expression of hGPR119 in Suspension-Adapted HEK 293FT Cells
HEK 293FT cells (Invitrogen, Carlsbad, CA) were grown in a shake flask in
293Freestyle medium (Invitrogen) supplemented with 50 microg/mL neomycin and
10mM HEPES (37C, 8% carbon dioxide, shaking). The cells were centrifuged
gently
(approximately 500xg, 10 minutes) and the pellet resuspended in a mixture of
Dulbecco's PBS(minus Mg++/-Ca++) supplemented with 18% fetal bovine serum
(Sigma Aldrich) and P1 virus such that the multiplicity of infection (M01) was
10 and the
final cell density was 1.3 x 106/mL (total volume 2.5 liters). The cells were
transferred
to a 5 liter Wave Bioreactor Wavebag (Wave Technologies, MA) and incubated for
4
hours at 27 degrees Celsius (17 rocks/min, 7 degrees platform angle); at the
end of the
incubation period, an equal volume(2.5 liters) of 293Freestyle medium
supplemented
with 30mM sodium butyrate (Sigma Aldrich) was added (final concentration = 15
mM),
and the cells were grown for 20 hours (37 degrees Celsius, 8% CO2 [0.2
liters/min}, 25
rocks/ minute, 7 degrees platform angle). Cells were harvested via
centrifugation
(3,000xg, 10 minutes), washed once on DPBS (minus Ca++/Mg++), resuspended in
0.25M sucrose, 25mM HEPES, 0.5mM EDTA, pH 7.4 and frozen at -80 degrees
Celsius.
Membrane Preparation for Radioliqand Binding Assays
The frozen cells were thawed on ice and centrifuged at 700 x g (1400 rpm) for
10
minutes at 4 degrees Celsius. The cell pellet was resuspended in 20 mL
phosphate-
buffered saline, and centrifuged at 1400 rpm for 10 minutes. The cell pellet
was then
resuspended in homogenization buffer (10 mM HEPES (Gibco #15630), pH 7.5, 1 mM
EDTA (BioSolutions, #610260-15), 1 mM EGTA (Sigma, #E-4378), 0.01 mg/mL
benzamidine (Sigma #13 6506), 0.01 mg/mL bacitracin (Sigma #13 0125), 0.005
mg/mL
leupeptin (Sigma #L 8511), 0.005 mg/mL aprotinin (Sigma #A 1153)) and
incubated on
ice for 10 minutes. Cells were then lysed with 15 gentle strokes of a tight-
fitting glass
Dounce homogenizer. The homogenate was centrifuged at 1000 x g (2200 rpm) for
10
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minutes at 4 degrees Celsius. The supernatant was transferred into fresh
centrifuge
tubes on ice. The cell pellet was resuspended in homogenization buffer, and
centrifuged again at 1000 x g (2200 rpm) for 10 minutes at 4 degrees Celsius
after
which the supernatant was removed and the pellet resuspended in homogenization
buffer. This process was repeated a third time, after which the supernatants
were
combined, Benzonase (Novagen # 71206) and MgC12 (Fluka #63020) were added to
final concentrations of 1 U/mL and 6 mM, respectively, and incubated on ice
for one
hour. The solution was then centrifuged at 25,000 x g (15000 rpm) for 20
minutes at 4
degrees Celsius, the supernatant was discarded, and the pellet was resuspended
in
fresh homogenization buffer (minus Benzonase and MgC12). After repeating the
25,000
x g centrifugation step, the final membrane pellet was resuspended in
homogenization
buffer and frozen at -80 degrees Celsius. The protein concentration was
determined
using the Pierce BCA protein assay kit (Pierce reagents A #23223 and B
#23224).
Synthesis and Purification of [3H1-Compound A
0 0
0
0
()-)
tritium gas NL-
N N 3
= Q H
+ z P PF6 3H
='I(\
Fy
SO2CH3 SO2C1-13 n
Compound A (Crabtree's catalyst) [3N-Compound A
CH2Cl2
Compound A ( isopropyl 4-(1-(4-(methylsulfonyl)phenyI)-3a,7a-dihydro-1H-
pyrazolo[3,4-
d]pyrimidin-4-yloxy)piperidine-1-carboxylate, as shown above) (4 mg, 0.009
mmol) was
dissolved in 0.5 mL of dichloromethane, and the resulting solution was treated
with (1,5-
cyclooctadiene)(pyridine)(tricyclohexylphosphine)-iridium(1)
hexaflurophosphate (J.
Organometal. Chem. 1979, 168, 183) (5 mg, 0.006 mmol). The reaction vessel was
sealed and the solution was stirred under an atmosphere of tritium gas for 17
hours.
The reaction solvent was removed under reduced pressure and the resulting
residue
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was dissolved in ethanol. Purification of crude [3N-Compound A was performed
by
preparative HPLC using the following conditions.
Column: Atlantis, 4.6 x 150mm, 5 rn
Mobil Phase A: water! acetonitrile / formic acid (98 / 2 / 0.1)
Mobil Phase B: acetonitrile
Gradient: Time % B
0.00 30.0
1.00 30.0
13.00 80.0
Run time: 16 min
Post time: 5 min
Flow Rate: 1.5 mL/minute
lnj. Volume: 20-50 I_
lnj. Solvent: DMSO
Detection: UV at 210 nm and 245 nm
The specific activity of purified [3N-Compound A was determined by mass
spectroscopy
to be 70 Ci/mmol.
Alternatively the binding assay can be performed with [3N-Compound B.
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Synthesis and Purification of [3N-Compound B
0 0
CD.) OC\j
tritium gas
N)N N \
LN.."-- NI ___________________________ .
k , 1\1
N N 3H
IP Q Q_
+ / P PF6
1 3H 411
r\ n
SO2CH3 SO2CH3
Py
Compound B (Crabtree's catalyst) [31-I]-Compound
B
CH2Cl2
Compound B (tert-butyl 4-(1-(4-(methylsulfonyl)phenyI)-1H-pyrazolo[3,4-
d]pyrimidin-4-
yloxy)piperidine-1-carboxylate, as shown above)(5 mg, 10.6 mol) was dissolved
in 1.0
mL of dichloromethane and the resulting solution was treated with Crabtree's
catalyst (5
mg, 6.2 mol). The reaction vessel was sealed and the solution was stirred
under an
atmosphere of tritium gas for 17 hours. The reaction solvent was removed under
reduced pressure and the resulting residue was dissolved in ethanol.
Purification of
crude [3N-Compound B was performed by silica gel flash column chromatography
eluting with 70% hexanes / 30% ethyl acetate, followed by silica gel flash
column
chromatography eluting with 60% petroleum ether / 40% ethyl acetate.
The specific activity of purified [3N-Compound B was determined by mass
spectroscopy
to be 57.8 Ci/mmol.
GPR119 Radioligand Binding Assay
Test compounds were serially diluted in 100% DMSO (J.T. Baker #922401). 2
microL of each dilution was added to appropriate wells of a 96-well plate
(each
concentration in triplicate). Unlabeled Compound A (or Compound B), at a final
concentration of 10 microM, was used to determine non-specific binding.
[31-1]-Compound A (or [3N-Compound B) was diluted in binding buffer (50 mM
Tris-HCI,
pH 7.5, (Sigma #T7443), 10 mM MgC12 (Fluke 63020), 1 mM EDTA (BioSolutions
#610260-15), 0.15% bovine serum albumin (Sigma #A7511 ), 0.01 mg/mL
benzamidine (Sigma #6 6506), 0.01 mg/mL bacitracin (Sigma #6 0125), 0.005
mg/mL
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leupeptin (Sigma #L 8511), 0.005 mg/mL aprotinin (Sigma #A 1153)) to a
concentration
of 60 nM, and 100 microL added to all wells of 96-well plate (Nalge Nunc
#267245).
Membranes expressing GPR119 were thawed and diluted to a final concentration
of 20
g/100 microL per well in Binding Buffer, and 100 microL of diluted membranes
were
added to each well of 96-well plate.
The plate was incubated for 60 minutes w/shaking at room temperature
(approximately
25 degrees Celsius). The assay was terminated by vacuum filtration onto GF/C
filter
plates (Packard #6005174) presoaked in 0.3% polyethylenamine, using a Packard
harvester. Filters were then washed six times using washing buffer (50 mM Tris-
HCI,
pH 7.5 kept at 4 degrees Celsius). The filter plates were then air-dyed at
room
temperature overnight. 30 I of scintillation fluid (Ready Safe, Beckman
Coulter
#141349) was added to each well, plates were sealed, and radioactivity
associated with
each filter was measured using a Wallac Trilux MicroBeta, plate-based
scintillation
counter.
The Kd for [3N-Compound A (or [31-1]-Compound B) was determined by carrying
out
saturation binding, with data analysis by non-linear regression, fit to a one-
site
hyperbola (Graph Pad Prism). IC50 determinations were made from competition
curves,
analyzed with a proprietary curve fitting program (SIGHTS) and a 4-parameter
logistic
dose response equation. Ki values were calculated from IC50 values, using the
Cheng-
Prusoff equation.
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The following results were obtained for the Beta-lactamase, Beta-arrestin,
cAMP, and
binding assays:
Human B- Human Human B-
lactamase Intrinsic cAMP Intrinsic arrestin Intrinsic Human
Example
Functional Activity* Functional Activity* Functional Activity*
N umber Binding
EC50 (%) EC50 (%) EC50 (%) Ki (nM)
(nM) (nM) (nM)
11 782 98.6 135 65.7 171 51 530
12 5200 100 10000 1700
13 142 91.3 217 119 149
14 464 75.7 495 109 723
15 135 120 83 89.2 134
16 220 108 445 93.7
17 31.5 93 161 67.4
18 68 77.3 56.7 71.2 153
19 4390 100 4510 99.5 6100
20 89.4 104 72.9 115 478 63.4 76.5
21 372 98.1 43.2 32.6 86.4
22 154 29 278 52.8 159
23 1550 100 150
24 141 103 43.6 124 62.9 107 99
25 79.5 101 192 92.8 418
26 217 60.1 264 73.9 270
27 2520 101 394 27 1050
28 3.57 24.6 10000 6100
29 3010 100 2550 33.2 2290
30 151 94.6 821 64.1 556 54.7 2020
31 54.1 108 155 91.8 462
32 281 86.5 62.6 68 206 81.8 514
33 258 88.4 1330 100
34 270 102 62.3 86 85 96.5 87.4
35 97.3 91.7 24.9 66.4 50.6 49 29.8
36 848 100 268 63.2 240 57.1 870
37 514 109 1160 55.9 416
38 22.1 122 15 99.8 29.8
39 10.7 76.8 5.32 86.1 14.9
40 10000 10000 1210
41 262 24.1 250 44.1 349
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42 120 84.3 147 86.1 223
43 2.78 75.5 5.36 82.5 9.25
44 2.13 79.8 45
45 47.1 51.8 102 58.2 352
46 30.2 102 13.5 91.6 44.8
47 52.7 130 32.8 106 150
48 51.4 113 71.7 111 200
49 24 66.3 4.11 113 18.5
50 19.3 120 30.2 97.6 36.5
51 40.7 56.1 142
52 10000 1440
53 194 83.8 668
54 233 115 594
55 456 22.5 361 46.5 939
56 245 36.7 164 53.4
57 2230 93.4 1520 84.5 4070
58 10000 6580
59 2070 96 1460 100 1380
60 7.4 77 18
61 156 47
62 152 104 393
63 226 31 509
64 6820 100** 4230
65 1050 88 187
66 2760 100** 226
67 52 70 53
68 10000
Values are reported as the geometric mean
1 1 1 1 1 1 1
*The intrinsic activity is the percent of maximal activity of the test
compound,
relative to the activity of a standard GPR119 agonist, 4-[[6-[(2-fluoro-4
methylsulfonylphenyl) amino]pyrimidin-4-yl]oxy]piperidine-1-carboxylic acid
isopropyl
ester (W02005121121), or (S)-1-methylcyclopropyl 4-(1-fluoro-2-(2-(2,3,6-
trifluorophenyl)acetamido)ethyl)piperidine-1-carboxylate (see Figure below),
at a final
concentration of 10 micromolar.
**the curve was extrapolated to 100% to calculate an EC50.
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Please note blank entry spaces denote that the test was not performed for that
Example.
Structure of (S)-1-methylcyclopropyl 4-(1-fluoro-2-(2-(2,3,6-
trifluorophenyl)acetamido)-
ethyl)piperidine-1-carboxylate***
(Abs)
0 1
F NA 0
0 F 0 F.õõ,..õ.õ---,,...)
N
H
F
***Presented at the SMASH workshop "NMR, It's Not Just For Structures:
Determination
of Physicochemical Properties" in Portland, Oregon on Tuesday September 28,
2010
The Gauche effect: Using Conformational Restriction of a Ethyl Amide Series to
Improve the Physical Properties of Analogues' by Kathleen Farley.
In Vivo Data
All in vivo protocols were approved by the Pfizer's Animal Welfare Committee.
Naive
male Wistar rats (200-250 g body weight on receipt) were obtained from Harlan
Laboratories (Indianapolis, IN), were pair housed in hanging plastic caging on
Sani-
chips sawdust bedding, and fed ad libitum on Purina 5001 chow. The rats were
housed
under a controlled light cycle (light from 6 am to 6 pm) at controlled
temperature and
humidity conditions. Rats were acclimated to the facility for at least 1 week
prior to
study.
Compound preparation
Example 50 was formulated as a 10% SDD in the vehicle 20 mM Tris Buffer at pH
7.4
with 0.5% methylcellulose and 0.5% HPMCAS-HF. The dose (75 mg/kg) was
formulated at 15 mg/mL for administration at 5 mL/kg, the required bulk was
added to a
mortar and ground with a small amount of vehicle to a smooth paste with a
pestle,
additional vehicle was added until the mixture flowed, when it was transferred
to a
stirred container, the mortar was rinsed several times with remaining quantity
of vehicle
and capped to prevent evaporation. The compound was formulated on the day of
doing
and was stirred continuously with a magnetic stir bar prior to, and during the
dosing
procedure.
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Oral glucose tolerance test (OGTT) protocol
Rats were stratified (n=8/group) to one of four dose groups 90 min or 30 min
pre-
glucose vehicle (20 mM Tris Buffer at pH 7.4 with 0.5% methylcellulose and
0.5%
Hydroxypropyl methylcellulose acetate succinate- high grade, fine particle
(HPMCAS-
HF), or 90 min or 30 min pre-glucose 75 mg/kg Example 50. Stratification was
performed according to body weight on day -1 to ensure that each group had
equal
group mean body weight values. The rats were fasted overnight in clean cages
(¨ 15
hours) prior to the oral glucose tolerance test. Body weights were recorded on
the
morning of the study (post fasting) for dose volume calculation. Blood samples
were
collected via the tail vein from all rats prior to dosing with vehicle or test
compound via
oral gavage (5 mL/kg). Ninety or thirty minutes later rats were bled and
immediately
dose with an oral dose of glucose (2 g/kg). The rats were re-bled at is, 30,
60 and 120
minutes post-glucose load. Blood samples (-250 microliter/time point) were
collected
into EDTA tubes with aprotinin/DPPIVi (0.6 TIU/20 microliter per mL whole
blood).
Blood tubes were inverted several times immediately following collection and
placed on
ice, then spun at 14,000 rpm in a refrigerated centrifuge for 5 minutes.
Plasma samples
were analyzed for glucose levels using a Roche c311 clinical chemistry
analyzer,
plasma insulin concentrations were determined using the Alpco Ultra-Sensitive
Insulin
Rat ELISA, and total amide GLP-1 concentrations were determined using MSD
ELISA
kit.
The results are presented as mean +/- SEM (standard error of the mean) unless
otherwise stated. Statistical evaluation of the data was carried out using one-
way
analysis of variance (ANOVA) with appropriate post-hoc analysis between time-
matched vehicle and treatment groups. Differences compared to vehicle with a
P<0.05
were considered statistically significant using Unadjusted T-test.
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Table 1: Effect of Example 50 during OGTT
Total Amide
Glucose 0-120 Insulin 0-60
GLP-1 0-120
Dose min AUC min AUC
min AUC
Dose Time (Example (percent (percent
(percent
50) vehicle vehicle
vehicle
response) response)
response)
90 min pre ++
75 mg/kg
glucose 90 82 103
30 min pre ++
75 mg/kg
glucose 89 94 153
++ p < 0.01 compared to time-matched vehicle
Preparation of Starting Materials
Preparation 1: Isopropyl 4-hydrazinopiperidine-1-carboxylate dihydrochloride
salt
H 0
H2N-N-( _________________________________________ \ N- ,
Isopropyl 4-{2-(tert-butoxycarbonyl)hydrazinyl}piperidine-1-carboxylate
(obtained as
described in W02008137436) (20.2 g, 67.02 mmol), was dissolved in absolute
ethanol
(250 mL), and the solution was stirred under nitrogen at room temperature.
Concentrated aqueous hydrochloric acid (27.9 mL, 335 mmol) was added slowly.
The
solution was stirred under nitrogen at room temperature for 4 hours. The
reaction was
concentrated to a white solid that contained some starting material. The solid
was
treated with a 4 M solution of hydrogen chloride in 1,4-dioxane (100 mL, 400
mmol) and
the resulting mixture was stirred for 14 hours at room temperature. The
reaction was
then concentrated under reduced pressure to give a white solid, which was
treated with
heptane (100 mL) and concentrated again to yield the title compound as a white
solid
(15 g, 81%). 1H NMR (400MHz, methanol-d4) delta 4.9 (m, 1 H), 4.1 (m, 2 H),
3.2 (m,
1H), 2.9 (m, 2 H), 2.0 (m, 2H), 1.4 (m, 2H), 1.2, (d, 6 H); LCMS (ES+): 202
(M+1).
Preparation 2: Isopropyl 4-1-5-amino-4-(ethoxycarbony1)-1H-pyrazol-1-yll-
piperidine-1-
carboxylate
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0
).,...._<N H2
A mixture of isopropyl 4-hydrazinopiperidine-1-carboxylate dihydrochloride
salt (7.08 g,
25.8 mmol), ethyl 2-cyano-3-ethoxyacrylate (4.81 g, 28.4 mmol), sodium acetate
(6.49 g,
77.5 mmol), and ethanol (80 mL) was stirred at 85 C for 3 hours. The mixture
was
concentrated to about a third of the initial volume. Water (50 mL), saturated
sodium
bicarbonate (50 mL), and brine (50 mL) were added. The resulting mixture was
extracted with ethyl acetate (2 x 50 mL). The combined organic extracts were
washed
with brine and dried over magnesium sulfate. The mixture was filtered, and the
filtrate
concentrated under vacuum to obtain the crude title compound as a light yellow
solid
(9.8 g), which was used in the next step without purification. An analytical
sample was
prepared by purification via chromatography on silica gel, eluting with a 30 %
to 60 %
solution of ethyl acetate in heptane. 1H NMR (500 MHz, deuterochloroform)
delta 1.26
(d, 6 H) 1.35 (t, 3 H) 1.86- 1.95 (m, 2 H) 2.04 - 2.17 (m, 2 H) 2.84 - 2.96
(m, 2 H) 3.89 -
3.98 (m, 1 H) 4.28 (q, 2 H) 4.25 - 4.40 (m, 2 H) 4.89 - 4.97 (m, 1 H) 5.06 (s,
2 H) 7.64 (s,
1 H); LCMS (ES+): 325.1 (M+1).
Preparation 3: Isopropyl 4-[5-bromo-4-(ethoxycarbonyI)-1H-pyrazol-1-
yl]piperidine-1-
carboxylate
0
yr
N / 0
Neat tert-butyl nitrite (4.8 mL, 39.3 mmol) was added slowly to a stirred
mixture of
isopropyl 4-[5-amino-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-piperidine-1-
carboxylate
(Preparation 2) (8.5 g, 26.2 mmol) and copper (II) bromide (3.7 g, 16 mmol) in
acetonitrile (100 mL) at room temperature. A significant exothermic effect was
observed with the mixture warming to about 50 C. After continued heating at
65 C for
minutes, the reaction was cooled to room temperature, and then concentrated
under
vacuum. An excess of 10 % aqueous ammonia was added, and the mixture was
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extracted with ethyl acetate. The organic phase was washed with water and
brine, and
concentrated under vacuum. The residue was purified by chromatography on
silica gel
eluting with 30 % to 70% ethyl acetate in heptane to provide the title
compound as a
yellow oil, which was about 70% pure by NMR and LCMS. The material was used in
the next step without further purification. 1H NMR (400 MHz,
deuterochloroform) delta
1.23(d, 6 H) 1.34(t, 3 H) 1.84- 1.95(m, 2 H) 2.01 - 2.15 (m, 2 H) 2.82 - 2.98
(m, 2 H)
4.25 - 4.36 (m, 2 H) 4.30 (q, 2 H) 4.45 - 4.56 (m, 1 H) 4.86 - 4.96 (m, 1 H)
7.95 (s, 1 H);
LCMS (ES+): 387.9 (M+1).
Preparation 4: Isopropyl 415-bromo-4-(hydroxymethyl)-1H-pyrazol-1-
yl]piperidine-1-
carboxylate
Br
HO\e_ _( ________________________________ \ p
, ,N
N _______________________________________ / To a solution of isopropyl 445-
bromo-4-(ethoxycarbony1)-1H-pyrazol-1-yl]piperidine-1-
carboxylate (3.59 g, 6.5 mmol) in tetrahydrofuran (32 mL) cooled to 0 C was
added a 2
M solution of borane-methyl sulfide complex in tetrahydrofuran (14.6 mL, 29.2
mmol).
The reaction mixture was heated at reflux for 21 hours and then stirred for 4
hours at
room temperature. The mixture was cooled to 0 C, and methanol was added. The
resulting solution was warmed to room temperature and stirred for 10 minutes.
The
solution was re-cooled to 0 C and aqueous 2 M sodium hydroxide solution (10
mL) was
added dropwise. The resulting mixture was diluted with ethyl acetate and
stirred
vigorously for 30 minutes. The layers were separated, and the aqueous phase
was
extracted twice with ethyl acetate. The combined organic layers were washed
sequentially with water and brine and then dried over magnesium sulfate. The
mixture
was filtered, and the filtrate concentrated under vacuum. Chromatography over
silica gel
eluting with 55% to 70% ethyl acetate in heptane gave the title compound as an
oil
(1.89 g, 84%). 1H NMR (400 MHz, deuterochloroform) delta 1.23 (d, 6 H), 1.87-
1.95
(br m, 3 H), 2.06 (qd, 2 H), 2.89 (br t, 2 H), 4.29 (br s, 2 H), 4.39 (tt, 1
H), 4.50 (d, 2 H),
4.90 (m, 1 H), 7.58 (s, 1 H); LCMS (ES+) 348.0 (M+1).
Preparation 5: Isopropyl 4-1-5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yllpiperidine-1-
carboxylate
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CN
HOc...7-(- _( ___________________________ \ p
,
N,N
/ 0¨
Isopropyl 4[5-bromo-4-(hydroxymethyl)-1H-pyrazol-1-yl]piperidine-1-
carboxylate (1.42
g, 4.10 mmol), tris-(dibenzylideneacetone) dipalladium (156 mg, 0.170 mmol), 1-
t-bis-
(diphenylphosphino) ferrocene (192 mg, 0.346 mmol), zinc dust (68.8 mg, 1.06
mmol),
zinc cyanide (497 mg, 4.23 mmol) and N,N-dimethylacetamide (20 mL) were
combined
in a microwave vial. The vial was flushed with nitrogen, sealed and heated at
120 C for
1 hour in a microwave reactor (Biotage Initiator 2.2). The reaction mixture
was passed
through a pad of FlorisilTM, diluted with ethyl acetate and then water was
added. The
aqueous phase was extracted 3 times with ethyl acetate and the combined
organic
layers were dried over magnesium sulfate. The mixture was filtered, and the
filtrate
evaporated under vacuum. Chromatography on silica gel eluting with 55% to 70%
ethyl
acetate in heptane gave the title compound as a green oil that solidified upon
standing
(1.06 g, 88%). 1H NMR (400 MHz, deuterochloroform) delta 1.24 (d, 6 H), 1.99
(br d, 2
H), 2.06 - 2.17 (m, 3 H), 2.93 (br t, 2 H), 4.31 (br s, 2 H), 4.48 (tt, 1 H),
4.71 (d, 2 H),
4.92 (m, 1 H), 7.60 (s, 1 H); LCMS (ES+): 293.1 (M+H).
Preparation 6: 2-Fluoro-4-1-(2-hydroxyethypthiOlPhend
F
401 OH
HOs
To a solution of 4-bromo-2-fluorophenol (0.75 mL, 6.8 mmol) and
diisopropylethylamine
(3.5 mL, 20.09 mmol) in 1,4-dioxane (35 mL) was added 9,9-dimethy1-4,5-
bis(diphenylphosphino)xanthene (415 mg, 0.717 mmol),
bis(dibenzylideneacetone)palladium (322 mg, 0.351 mmol) and 2-mercaptoethanol
(0.46 mL, 6.86 mmol), and the dark brown reaction solution was heated at 110
C for
16 hours. The reaction was allowed to cool to room temperature, diluted with
water and
extracted with ethyl acetate four times. The organic extracts were combined
and dried
over magnesium sulfate, The mixture was filtered, and the filtrate
concentrated under
reduced pressure to give a maroon oil which was purified by chromatography on
silicon
gel to afford the title compound (985 mg, 76 A) as a maroon solid. 1H NMR
(400 MHz,
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deuterochloroform) delta 3.00 (t, 2H, J=5.95 Hz) 3.69 (d, 2 H, J=3.71 Hz) 6.89-
6.95 (m,
1 H) 7.11 (ddd, 1 H, J=8.39, 2.15, 1.17 Hz) 7.17 (dd, 1 H, J=10.54, 2.15 Hz).
Preparation 7: 4-[(2-{[tert-Butyl(dimethyl)silyl]oxylethyl)thio]-2-
fluorophenol
401 OH
/ \
To a solution of 2-fluoro-4-[(2-hydroxyethypthio]phenol (985 mg, 5.24 mmol)
and
imidazole (371 mg, 5.30 mmol) in N,N-dimethylformamide (5 mL) was added tert-
butyldimethylsilyl chloride (814 mg, 5.24 mmol) portion-wise, and the reaction
was
stirred at room temperature for 4 hours. The reaction was concentrated under
reduced
pressure, and the residue diluted with water followed by extraction with ethyl
acetate
three times. The combined organic extracts were washed with brine and dried
over
magnesium sulfate. The mixture was filtered, and the filtrate concentrated
under
reduced pressure to give the title compound as an orange oil (1.43 g, 90 %)
which was
used without further purification. LCMS (ES+): 301.1 (M-1).
Preparation 8: 1-1-4-(Benzyloxy)-3-fluorophenyll-1H-tetrazole
0
To a suspension of 4-(benzyloxy)-3-fluoroaniline (1.04 g, 4.8 mmol) (WO
2005030140)
under a nitrogen atmosphere was added acetic acid (2.3 mL, 38.3 mmol), triethy
lorthoformate (2.44 mL, 14.4 mmol) and sodium azide (0.34 g, 5.3 mmol), and
the
reaction mixture heated at 95 C for 2.5 hours. The solution was then allowed
to cool to
room temperature, and water was added followed by extraction with ethyl
acetate three
times. The extracts were combined and washed with brine and dried over
magnesium
sulfate. The mixture was filtered and concentrated under reduced pressure, and
the
crude material purified by chromatography on silicon gel (20 ¨ 40 % ethyl
acetate in
heptane) to give the title compound as a white solid (1.12 g, 86%). 1H NMR
(400 MHz,
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deuteromethanol) delta 9.65 (s, 1H), 7.73 - 7.68 (dd, 1H, J=11, 2.5 Hz), 7.60 -
7.57 (m,
1H) 7.47 - 7.45 (m, 2H), 7.40 - 7.30 (m, 5H), 5.24 (s, 2H); LCMS (ES+): 271.1
(M+1).
Preparation 9: 2-Fluoro-4-(1H-tetrazol-1-yl)phenol
N%-\ =
,,... ,N
"Thl OH
F
To 1[4-(benzyloxy)-3-fluoropheny1]-1H-tetrazole (1.12 g, 4.14 mmol) in a Parr
shaker
flask was added ethanol (40 mL), and the solution purged with nitrogen gas.
10%
palladium on carbon (0.30 g) was added, and the reaction hydrogenated on a
Parr
shaker apparatus at 40 psi of hydrogen for 30 minutes. The mixture was then
filtered
through a micro pore filter, and the filtrate was concentrated under reduced
pressure to
yield the title compound as a white solid (0.67 g, 90 `)/0) which was use
without
purification. 1H NMR (400 MHz, deuteromethanol) delta 9.62 (s, 1H), 7.65 -
7.62 (dd,
1H, J=11, 2.5 Hz), 7.50 - 7.46 (m, 1H) 7.47 - 7.45 (dd, 1H, J=9.0, 9.0 Hz);
LCMS
(ES+): 181.1 (M+1).
Preparation 10: Isopropyl 4-(5-cyano-4-((methylsulfonyloxy)methyl)-1H-pyrazol-
1-
y1)piperidine-1-carboxylate
N
/ \\CI
Isopropyl 4-(5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-y1)piperidine-1-
carboxylate
(Preparation 5) (75 mg, 0.24 mmol) was dissolved in 1 mL of anhydrous
dichloromethane and triethylamine (0.1 mL, 0.74 mmol) was added. The reaction
mixture was cooled in an ice bath and methanesulfonic anhydride (62 mg, 0.34
mmol)
was then added. The solution was removed from the ice bath and stirred for 30
minutes.
The reaction was quenched by addition of saturated aqueous sodium bicarbonate
and
the layers were separated. The aqueous layer was extracted three more times
with
dichloromethane. The organic extracts were combined and washed with brine,
dried
over sodium sulfate, filtered and the filtrate was concentrated to give an oil
(75 mg,
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100% yield). The crude material was used in subsequent steps without further
purification.
Preparation 11: tert-Butyl 4-hydrazinopiperidine-1-carboxylate hydrochloride
salt
H/\ 0
H2N-N- N-
/
HCI 0 <
Into a solution of tert-butyl 4-oxopiperidine-1-carboxylate (50 g, 0.25 mmol)
in methanol
(500 mL) in an autoclave was added hydrazine mono-hydrochloride (17.2 g, 0.25
mmol)
in water (100 mL). The white mixture was stirred under argon followed by the
addition
of 5% platinum on carbon (750 mg) as a slurry in water. The autoclave was
sealed and
charged to 60 atmospheres with hydrogen, and the reaction was stirred for 15
hours.
Upon completion, the reaction was filtered through Celite , and the pad washed
with
methanol. This preparation was carried out six times. The combined filtrates
were
concentrated under reduced pressure, and the resulting white precipitate (di-
tert-butyl-
4,4'-hydrazine-1,2-diyldipiperidine-1-carboxylate) by-product was collected by
filtration
and washed several times with water. The aqueous filtrate was then
concentrated under
reduced pressure to give the desired product (221 g, 59%) as a colorless
solid. 1H NMR
(400MHz, deuterochloroform) delta 4.13 (br s, 2H), 3.32 (br t, 1H), 2.77 (br
t, 2H), 2.16
(m, 2H), 1.66 (m, 2H), 1.43 (s, 9H).
Preparation 12: tert-Butyl 4-1-5-amino-4-(ethoxycarbony1)-1H-pyrazol-1-
yllpiperidine-1-
carboxylate
0
yH2
N / 0 <
A mixture of tert-butyl 4-hydrazinopiperidine-1-carboxylate hydrochloride salt
(221 g,
880 mmol), ethyl 2-cyano-3-ethoxyacrylate (153 g, 880 mmol), sodium acetate
trihydrate (477 g, 352 mmol) and ethanol (2000 mL) was stirred at 85 degrees
Celsius
for 8 hours. The mixture was concentrated under reduced pressure, and the
residue
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dissolved in ethyl acetate and water. The layers were separated, and the
aqueous layer
extracted with ethyl acetate. The combined organic extracts were then dried
over
magnesium sulfate. The mixture was filtered, and the filtrate concentrated
under
reduced pressure. The crude material was purified by filtration through a
short plug of
silica gel eluting with 40% ethyl acetate in heptane to produce the product as
a pale
yellow solid (214 g, 72%). 1H NMR (500 MHz, deuterochloroform) delta 7.60 (s,
1H),
5.27 (br s, 2H), 4.23 (br q, 4H), 3.91 (m, 1H), 2.81 (br s, 2H), 2.04 (m, 2H),
1.86 (m, 2H),
1.44 (s, 9H), 1.29 (t, 3H).
Preparation 13: tert-Butyl 4-[5-bromo-4-(ethoxycarbonyI)-1H-pyrazol-1-
yl]piperidine-1-
carboxylate
ci
)Lir
00
.L7:N-( \.N1- ,
N / 0 C
To a solution of copper (II) bromide (1.69 g, 770 mmol) in acetonitrile (1000
mL) was
slowly added tert-butyl nitrite (112 mL, 960 mmol), and the solution was
heated to 65
degrees Celsius. To this was added a solution of tert-butyl 4-[5-amino-4-
(ethoxycarbony1)-1H-pyrazol-1-yl]piperidine-1-carboxylate (215 g, 640 mmol) in
acetonitrile (650 mL) drop-wise over 30 minutes. After 4 hours, the reaction
was
allowed to cool to room temperature and was then poured into 2 M hydrochloric
acid
(1500 mL) in ice. The mixture was extracted with ethyl acetate three times,
and the
combined organic extracts were washed with saturated aqueous sodium
bicarbonate
and then dried over magnesium sulfate. The mixture was filtered, and the
filtrate
concentrated under reduced pressure. The resulting residue was purified by
filtration
through a short plug of silica gel eluting initially with 10% heptane in
dichloromethane
followed by dichloromethane to give the title compound (137 g, 53%) as a
yellow oil
which solidified on standing. 1H NMR (400 MHz, deuterochloroform) delta 7.95
(s, 1H),
4.48 (m, 1H), 4.28 (br q, 4H), 2.86 (br s, 2H), 2.06 (m, 2H), 1.90 (m, 2H),
1.44 (s, 9H),
1.34 (t, 3H).
Preparation 14: tert-Butyl 415-bromo-4-(hydroxymethyl)-1H-pyrazol-1-
yl]piperidine-1-
carboxylate
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Br
0
,N
N __________________________________________ 0 <
To a solution of tert-butyl 4-[5-bromo-4-(ethoxycarbonyI)-1H-pyrazol-1-
yl]piperidine-1-
carboxylate (137 g, 0.34 mol) in tetrahydrofuran (1300 mL) cooled to 0 degrees
Celsius
was slowly added borane-methyl sulfide (97 mL, 1.02 mol). The solution was
allowed to
warm to room temperature and then heated at reflux for 15 hours. The reaction
was
then cooled in an ice bath, and methanol (40 mL) added drop-wise. The solution
was
then stirred at room temperature for 20 minutes. Aqueous 2 M sodium hydroxide
(1200
mL) was added, and the layers were separated. The aqueous layer was extracted
with
ethyl acetate, and the combined organics layers were washed with brine, dried
over
magnesium sulfate, and the solvent removed under reduced pressure. The
resulting
residue was purified by filtration through a short plug of silica gel eluting
with 30% ethyl
acetate in heptane to reveal the title compound as an colorless solid (61.4 g,
50%).
Impure material from this purification was further purified via the above
chromatographic
procedure to provide a second batch of the title compound (22 g, 18%) as a
colorless
solid. 1H NMR (400 MHz, deuterochloroform) delta 7.59 (s, 1H), 4.52 (s, 2H),
4.37 (m,
1H), 4.25 (br s, 2H), 2.86 (br s, 2H), 2.06 (m br s, 2H), 1.89 (m, 2H), 1.45
(s, 9H).
Preparation 15: tert-Butyl 445-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yllpiperidine-1-
carboxylate
9/
\ o(N
=/
__________________________________ ' 0
Copper (I) cyanide (2.97 g, 33.3 mmol) was added to a stirred solution of tert-
butyl 445-
bromo-4-(hydroxymethyl)-1H-pyrazol-1-yl]piperidine-1-carboxylate (10 g, 27.8
mmol) in
degassed dimethylformamide (100 mL). The reaction was then heated at 165
degrees
Celsius for 4 hours and allowed to cool to room temperature. It was further
cooled in an
ice-bath, and a solution of ethylenediamine (5.5 mL) in water (20 mL) was
added
followed by dilution with more water (70 mL). The mixture was then extracted
with ethyl
acetate, and the layers separated. The organic layer was washed sequentially
with
water and brine and then dried over magnesium sulfate. The mixture was
filtered and
the filtrate concentrated under reduced pressure. This procedure was carried
out in 8
batches. The final crude residues were combined and purified by repeated
silica gel
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column chromatography eluting with 40 % ethyl acetate in heptane to give the
title
compound (11.6 g, 17%) as a colorless solid. 1H NMR (400MHz,
deuterochloroform)
7.59 (s, 1H), 4.71 (s, 2H), 4.45 (m, 1H), 4.26 (br s, 2H), 2.88 (br t, 2H),
2.08 (m, 2H),
1.98 (m, 2H), 1.48 (s, 9H); LCMS (ES+): 207.1 (M-Boc+H).
For an alternative synthesis of tert-butyl 445-cyano-4-(hydroxymethyl)-1H-
pyrazol-1-
yl]piperidine-1-carboxylate please see Example 50.
Preparation 16: tert-Butyl 4-(5-cyano-4-((methylsulfonyloxy)methyl)-1H-pyrazol-
1-
yl)piperidine-1-carboxylate
0 1/1
0 (0/
0
To a stirred solution of tert-butyl 4-(5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yl)piperidine-1-carboxylate (202 mg, 0.659 mmol) in dichloromethane (6.6 mL)
was
added triethylamine (0.18 mL, 1.32 mmol) followed by methanesulfonic anhydride
(189
mg, 1.1 mmol) at room temperature. The mixture was stirred for 4.5 hours
before it was
diluted with dichloromethane and saturated aqueous bicarbonate. The layers
were
separated and the aqueous layer was extracted with dichloromethane. The
combined
organic extracts were washed with brine, dried over magnesium sulfate,
filtered and the
filtrate was concentrated in vacuo to give tert-butyl 4-(5-cyano-4-
((methylsulfonyloxy)methyl)-1H-pyrazol-1-y1)piperidine-1-carboxylate as an oil
which
was used without further purification.
Preparation 17: 2-Fluoro-4-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-tetrazol-5-
yl)phenol
and 2-Fluoro-4-(2-((2-(trimethylsilypethoxy)methyl)-2H-tetrazol-5-y1)phenol
N-1\!
N=N
s,N
NL
40/ N 0¨\_s(
HO 0
HO
Si
A) 4-(Benzyloxy)-3-fluorobenzonitrile
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To a stirred solution of 3-fluoro-4-hydroxybenzonitrile (1.00 g, 7.30 mmol) in
20 mL of
acetonitrile was added portion-wise potassium carbonate (2.02 g, 14.6 mmol).
The
resulting mixture was stirred for 10 minutes before benzyl bromide (1.33 mL,
10.9
mmol) was added. The mixture was stirred at room temperature for 70 hours
before it
was diluted with ethyl acetate and water. The organic phase was separated and
washed
with water, brine, dried over magnesium sulfate, filtered and the filtrate was
concentrated in vacuo. The residue was purified by flash chromatography,
eluting with a
gradient of 5 to 20% of ethyl acetate in heptane to give 4-(benzyloxy)-3-
fluorobenzonitrile as a white solid (1.33 g).
B) 5-(4-(Benzyloxy)-3-fluorophenyI)-1H-tetrazole and 5-(4-(Benzyloxy)-3-
fluorophenyI)-
2H-tetrazole
A vial charged with 4-(benzyloxy)-3-fluorobenzonitrile (250 mg, 1.10 mmol),
sodium
azide (214 mg, 3.30 mmol), ammonium chloride (176 mg, 3.30 mmol) and 3 mL of
N,N-
dimethylformamide was heated at 110 degrees Celsius for 18 hours. The reaction
mixture was cooled to room temperature, diluted with water and ethyl acetate
and the
pH was adjusted to 3 using aqueous 1 N hydrochloric acid. The organic phase
was
separated and washed with brine, dried over magnesium sulfate, filtered and
the filtrate
was concentrated in vacuo to give the title compounds as a white solid (270
mg). This
material was used in subsequent steps without purification.
C) 5-(4-(Benzyloxy)-3-fluoropheny1)-1-((2-(trimethylsilypethoxy)methyl)-1H-
tetrazole and
5-(4-(Benzyloxy)-3-fluoropheny1)-2-((2-(trimethylsilypethoxy)methyl)-2H-
tetrazole
To a solution of 5-(4-(benzyloxy)-3-fluorophenyI)-1H-tetrazole and 5-(4-
(benzyloxy)-3-
fluorophenyI)-2H-tetrazole (270 mg, 1 mmol) dissolved in tetrahydrofuran was
added
sodium hydride (44 mg, 1.1 mmol) in four portions and the resulting mixture
was stirred
at room temperature for 15 minutes. (2-(Chloromethoxy)ethyl)trimethylsilane
(0.19 mL,
1.0 mmol) was then added and the reaction mixture was stirred at room
temperature for
16 hours. The reaction was quenched by the addition of water and ethyl acetate
was
added. The organic phase was separated and the aqueous phase was extracted
twice
with ethyl acetate. The combined organic extracts were washed with brine,
dried over
magnesium sulfate, filtered and the filtrate was concentrated under reduced
pressure.
Purification by flash chromatography, eluting with a gradient of ethyl acetate
and
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heptane (5 to 20% ethyl acetate) gave the desired product as a white solid
(270 mg,
67% yield).
D) 2-Fluoro-4-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-tetrazol-5-yl)phenol
and 2-Fluoro-
4-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)phenol
To a solution of 5-(4-(benzyloxy)-3-fluoropheny1)-1-((2-
(trimethylsilypethoxy)methyl)-1H-
tetrazole and 5-(4-(benzyloxy)-3-fluoropheny1)-2-((2-
(trimethylsilypethoxy)methyl)-2H-
tetrazole (140 mg, 0.35 mmol) dissolved in a mixture of 2 mL of ethanol and 2
mL of
tetrahydrofuran was added palladium black (56 mg, 0.53 mmol) and formic acid
(0.14
mL, 3.5 mmol). The resulting mixture was stirred at room temperature for 4
hours before
being filtered though a pad of Celite . The filtrate was concentrated under
reduced
pressure and the resulting crude material was used in the subsequent step
without
further purification.
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Preparation 18: 5-(4-(Benzyloxy)-3-fluoropheny1)-1-(2-
(trimethylsilyloxy)ethyl)-1H-
tetrazole and 5-(4-(Benzyloxy)-3-fluoropheny1)-2-(2-(trimethylsilyloxy)ethyl)-
2H-tetrazole
N-N,
I ',N Nr---N,
N
0 NH
0 100
lei
F 0 I
'Si 0 0
F /
1
To a solution of 5-(4-(benzyloxy)-3-fluorophenyI)-1H-tetrazole and 5-(4-
(Benzyloxy)-3-
fluorophenyI)-2H-tetrazole (Preparation 17, Step B) (550 mg, 2 mmol) dissolved
in N,N-
dimethylformamide (8 mL) was added sodium hydride (163 mg, 4 mmol) in two
portions
and the resulting mixture was stirred at room temperature for 5 minutes. (2-
Bromoethoxy)trimethylsilane (1.3 mL, 6 mmol) was then added and the reaction
mixture
was stirred at 70 degrees Celsius for 16 hours before being cooled to room
temperature. The reaction was quenched by addition of water and ethyl acetate
was
added. The organic phase was separated and the aqueous phase was extracted
twice
with ethyl acetate. The combined organic extracts were washed with brine,
dried over
magnesium sulfate, filtered and concentrated under reduced pressure. The
residue was
purified by flash chromatography, eluting with a gradient of ethyl acetate and
heptane (5
to 30% ethyl acetate) to give 5-(4-(benzyloxy)-3-fluoropheny1)-1-(2-
(trimethylsilyloxy)ethyl)-1H-tetrazole (100 mg, 12% yield) and 5-(4-
(benzyloxy)-3-
fluoropheny1)-2-(2-(trimethylsilyloxy)ethyl)-2H-tetrazole (600 mg, 69% yield).
Preparation 19: 2-Fluoro-4-(2-(2-(trimethylsilyloxy)ethyl)-2H-tetrazol-5-
yl)phenol
N=N,
0
Si'
/
HO'
F
To a solution of 5-(4-(benzyloxy)-3-fluoropheny1)-2-(2-
(trimethylsilyloxy)ethyl)-2H-
tetrazole (Preparation 18)(230 mg, 0.54 mmol) dissolved in a mixture of 6 mL
of ethanol
and 6 mL of tetrahydrofuran was added palladium black (86 mg, 0.806 mmol) and
formic acid (0.215 mL, 5.4 mmol). The resulting mixture was stirred at room
temperature
for 4 hours before being filtered through a pad of Celite . The filtrate was
concentrated
under reduced pressure and the resulting crude material (180 mg) was used in
the
subsequent step without further purification.
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Preparation 20: 2-Fluoro-4-(1-(2-(trimethylsilyloxy)ethyl)-1H-tetrazol-5-
yl)phenol
N¨N,
I s,N
HO NH
F 0, I
Si
i
To a solution of 5-(4-(benzyloxy)-3-fluoropheny1)-1-(2-
(trimethylsilyloxy)ethyl)-1H-
tetrazole (Preparation 18)(130 mg, 0.30 mmol) dissolved in a mixture of 2 mL
of ethanol
and 2 mL of tetrahydrofuran was added palladium black (48 mg, 0.45 mmol) and
formic
acid (0.12 mL, 3 mmol). The resulting mixture was stirred at room temperature
for 4
hours before being filtered over a pad of Celite . The filtrate was
concentrated under
reduced pressure and the resulting crude material (94 mg) was used in the
subsequent
step without further purification.
Preparation 21: 2-Fluoro-4-(1-methy1-1H-tetrazol-5-y1)phenol
N-N,
I µ,N
ON
Ho
F
A) 5-(4-(Benzyloxy)-3-fluoropheny1)-1-methy1-1H-tetrazole and 5-(4-(Benzyloxy)-
3-
fluoropheny1)-2-methy1-2H-tetrazole
To a stirred solution of 5-(4-(benzyloxy)-3-fluorophenyI)-1H-tetrazole and 5-
(4-
(benzyloxy)-3-fluoropheny1)-2H-tetrazole (Preparation 17, Step B) (1.50 g,
5.55 mmol) in
30 mL of tetrahydrofuran was added sodium hydride (444 mg, 11.1 mmol) in two
portions at room temperature. After 5 minutes, iodomethane (1.04 mL, 16.6
mmol) was
added and the reaction was stirred under a nitrogen atmosphere for 15 hours at
room
temperature. The mixture was diluted with water and extracted twice with ethyl
acetate.
The combined organic extracts were washed with brine, dried with magnesium
sulfate,
filtered and the filtrate was concentrated in vacuo. The residue was purified
by flash
chromatography, eluting with 10-40% ethyl acetate in heptane to give 5-(4-
(benzyloxy)-
3-fluoropheny1)-2-methy1-2H-tetrazole as a white solid (1.1 g) and 5-(4-
(benzyloxy)-3-
fluoropheny1)-1-methyl-1H-tetrazole as a white solid (450 mg).
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5-(4-(Benzyloxy)-3-fluorophenyI)-1-methyl-1H-tetrazole. 1H NMR (400 MHz,
deuterochloroform) delta 4.15 (s, 3 H) 5.23 (s, 2 H) 7.15 (t, J=8.39 Hz, 1 H)
7.31 -7.48
(m, 6 H) 7.52 (dd, J=11.13, 2.15 Hz, 1 H). LCMS (M+1) 285.1.
B) 2-Fluoro-4-(1-methyl-1H-tetrazol-5-yl)phenol
To a solution of 5-(4-(benzyloxy)-3-fluorophenyI)-1-methyl-1H-tetrazole (500
mg, 1.76
mmol) in 6 mL of ethanol and 6 mL of tetrahydrofuran was added formic acid
(0.7 mL,
17.6 mmol) followed by palladium black (28i mg, 2.64 mmol). The reaction
mixture was
stirred at room temperature for 4 hours. The reaction mixture was filtered
through
Celite and the filtrate was concentrated in vacuo to give 2-fluoro-4-(1-
methyl-1H-
tetrazol-5-yl)phenol as a white solid (330 mg) which was used for in
subsequent
reactions without further purification.
Preparation 22: 4-(1-Methyl-1H-tetrazol-5-yl)bhenol
N-N,
I µ,N
ONI
HO
A) 4-(Benzyloxy)-N-methylbenzamide
To a flask charged with thionyl chloride (0.35 mL, 4.82 mmol) was added a
solution of
commercially available 4-benzyloxybenzoic acid (1.00 g, 4.38 mmol) in 10 mL of
dichloromethane and 0.01 mL of N,N-dimethylformamide at zero degrees Celsius
with
stirring. The ice bath was removed and the solution was stirred for 4 hours at
room
temperature. The mixture was concentrated in vacuo to give a white solid. This
solid
was taken up in 10 mL of methyl amine (2 M in tetrahydrofuran) and the
resulting
solution was stirred at room temperature for 70 hours. The mixture was diluted
with
ethyl acetate and water and the organic layer was separated, dried over
magnesium
sulfate, filtered and the filtrate was concentrated in vacuo to give a white
solid. This
solid was recrystallized from ethyl acetate and heptane to give 4-(benzyloxy)-
N-
methylbenzamide as white needles (850 mg).
B) 5-(4-(Benzyloxy)phenyI)-1-methyl-1H-tetrazole
To a stirred solution of 4-(benzyloxy)-N-methylbenzamide (200 mg, 0.829 mmol)
in 3
mL of acetonitrile and one drop of N,N-dimethylformamide, in a flask topped
with a
reflux condenser, was added triethylamine (0.12 mL) under a nitrogen
atmosphere.
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The reaction mixture was stirred for 10 minutes before thionyl chloride (0.078
mL, 1.08
mmol) was added drop-wise. The yellow reaction mixture was stirred for 1 hour
at room
temperature under a nitrogen atmosphere. Triethylamine (0.36 mL) was then
added
slowly, followed by tetrabutylammonium chloride (37.4 mg, 0.12 mmol) and
sodium
azide (611 mg, 1.82 mmol). The resulting yellow suspension was vigorously
stirred for
70 hours at room temperature under a nitrogen atmosphere. The mixture was
diluted
with water and ethyl acetate. The organic layer was separated, washed with
brine,
dried over magnesium sulfate, filtered and the filtrate was concentrated in
vacuo. The
residue was purified by flash chromatography, eluting with a gradient of 10 to
40% ethyl
acetate in heptane to give 5-(4-(benzyloxy)phenyI)-1-methyl-1H-tetrazole as a
white
solid (180 mg).
C) 4-(1-Methyl-1H-tetrazol-5-yl)phenol
To a stirred solution 5-(4-(benzyloxy)phenyI)-1-methyl-1H-tetrazole (180 mg,
0.676
mmol) in 3 mL of ethanol and 3 mL of tetrahydrofuran was added formic acid
(0.27 mL,
6.76 mmol) followed by palladium black (108 mg, 1.01mmol). The mixture was
stirred at
room temperature for 4 hours. The reaction mixture was filtered through Celite
and the
filtrate was concentrated to give 4-(1-methyl-1H-tetrazol-5-yl)phenol as a
white solid
(110 mg), which was used in subsequent reactions without further purification.
Preparation 23: 3-Fluoro-4-hydroxybenzamide
NH2
0
ilk F
OH
A mixture of commercially available 3-fluoro-4-hydroxybenzonitrile (500 mg,
3.65 mmol)
and potassium hydroxide (1.02 g, 18.2 mmol) in 10 mL of 80% ethanol was heated
at
reflux for 16 hours. After cooling to room temperature the mixture was
concentrated in
vacuo and the residue was taken up into water, acidified with acetic acid and
extracted
with ethyl acetate. The combined organic extracts were dried over magnesium
sulfate,
filtered and the filtrate was concentrated in vacuo. The residue was purified
by flash
chromatography, eluting with a gradient of 20 to 60% ethyl acetate in heptane
to give 3-
fluoro-4-hydroxybenzamide as a white solid (210 mg).
Alternatively, 3-fluoro-4-hydroxybenzamide can be prepared as follows:
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To a stirred solution of urea hydrogen peroxide (4.2 g, 43.8 mmol) in 12 mL of
water
was added solid sodium hydroxide (1.04 g, 25.5 mmol). The resulting solution
was
cooled in an ice bath before a solution of 3-fluoro-4-hydroxybenzonitrile
(1.00 g, 7.29
mmol) in 5 mL of ethanol was added. The mixture was vigorously stirred for 2
hours at
room temperature before it was diluted with water (100 mL) and ethyl acetate
(100 mL).
The mixture was stirred for 5 minutes before 1 M hydrochloric acid was added
until pH
4. The aqueous layer was separated and extracted with ethyl acetate (3X100
mL). The
combined organic layers were dried over magnesium sulfate, filtered, and the
filtrate
was concentrated to give a white solid. This solid was triturated with diethyl
ether and
heptane (2:1, 90 mL) for 1 hour, before filtering to give 3-fluoro-4-
hydroxybenzamide as
a white solid (1.05 g). 1H NMR (400 MHz, deutero dimethyl sulfoxide ) delta
6.93 (t,
J=8.69 Hz, 1 H) 7.19 (br. s., 1 H) 7.53 (dd, J=8.39, 1.95 Hz, 1 H) 7.62 (dd,
J=12.40,
2.05 Hz, 1 H) 7.77 (br. s., 1 H) 10.39 (s, 1 H). LCMS (ES) 156.0 (M+1).
Preparation 24: 2-Fluoro-4-hydroxybenzamide
NH2 F
0
lk
OH
To a stirred solution of urea hydrogen peroxide (2.1 g, 21.9 mmol) in 6 mL of
water was
added solid sodium hydroxide (521 mg, 12.8 mmol). The resulting solution was
cooled
in an ice bath before a solution of 2-fluoro-4-hydroxybenzonitrile (500 mg,
3.65 mmol) in
2 mL of ethanol was added. The mixture was vigorously stirred for 2 hours at
room
temperature before it was diluted with water (100 mL) and ethyl acetate (100
mL). The
mixture was stirred for 5 minutes before 1 M hydrochloric acid was added until
pH=4.
The aqueous layer was separated and extracted with ethyl acetate (3X50 mL).
The
combined organic layers were dried over magnesium sulfate, filtered, and the
filtrate
was concentrated to give 2-fluoro-4-hydroxybenzamide as a white solid.
Preparation 25: Isopropyl 4-(5-cyano-4-(1-hydroxyethyl)-1H-pyrazol-1-
yl)piperidine-1-
carboxylate
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N
--
------N' 0
Isopropyl 4-(5-cyano-4-formy1-1H-pyrazol-1-yl)piperidine-1-carboxylate
(Example 9,
Step A) (51 mg, 0.18 mmol) was dissolved in 2 mL of anhydrous tetrahydrofuran
and
cooled to negative 78 degrees Celsius under a nitrogen atmosphere.
Methylmagnesium
bromide (0.070 mL, 0.21 mmol, 3 M in diethyl ether) was then added drop-wise.
The
cold bath was removed and the mixture was stirred for 1 hour at room
temperature. The
mixture was diluted with 1 M aqueous potassium bisulfate and extracted three
times
with ethyl acetate. The combined organic extracts were washed with brine,
dried over
sodium sulfate, filtered and the filtrate was concentrated in vacuo. The
residue was
purified by flash chromatography, eluting with a gradient of ethyl acetate in
heptane (20
to 100% ethyl acetate) to give isopropyl 4-(5-cyano-4-(1-hydroxyethyl)-1H-
pyrazol-1-
y1)piperidine-1-carboxylate as a white solid (33 mg) which was used in
subsequent
steps without purification.
Preparation 26: 1-Methylcycloorooyl 4-nitroohenyl carbonate
0
)-0
02N 411 0
A) 1-Methylcyclopropanol
A 1 L flask was charged with titanium methoxide (100 g), cyclohexanol (232 g),
and
toluene (461 mL). The flask was equipped with a Dean-Stark trap and condenser.
The
mixture was heated at 140 degrees Celsius until the methanol was removed. The
toluene was removed at 180 degrees Celsius. More toluene was added and this
process was repeated twice. After all the toluene was removed the flask was
dried
under high vacuum. Diethyl ether (580 mL) was added to the flask to prepare a
1 M
solution in diethyl ether. A 5 L, 3-neck flask was equipped with an overhead
stirrer, inert
gas inlet and a pressure-equalizing addition funnel. The flask was flushed
with nitrogen
gas and charged with methyl acetate (60.1 mL, 756 mmol), titanium
cyclohexyloxide (1
M solution in ether 75.6 mL), and diethyl ether (1500 mL). The solution was
stirred while
keeping the reaction flask in a room temperature water bath. The addition
funnel was
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charged with the 3 M ethylmagnesium bromide solution (554 mL, 1.66 moles). The
Grignard reagent was added drop-wise over 3 hours at room temperature. The
mixture
became a light yellow solution, and then gradually a precipitate formed which
eventually
turned to a dark green/brown/black colored mixture. After stirring for an
additional 15
minutes, following the addition of the Grignard, the mixture was carefully
poured into a
mixture of 10% concentrated sulfuric acid in 1 L of water. The resulting
mixture was
stirred until all the solids dissolved. The aqueous layer was separated and
extracted
with diethyl ether 2 x 500 mL. The combined organic extracts were washed
sequentially
with water, brine, dried over potassium carbonate (500 g) for 30 minutes,
filtered and
the filtrate was concentrated in vacuo to an oil. Sodium bicarbonate (200 mg)
was
added and the crude material was distilled, collecting fractions boiling
around 100
degrees Celsius to give the title compound (23 grams) with methyl ethyl ketone
and 2-
butanol as minor impurities. 1H NMR (500 MHz, deuterochloroform) delta 0.45
(app. t,
J=6.59 Hz, 2 H), 0.77 (app. t, J=5.61 Hz, 2 H), 1.46 (s, 3 H). The preparation
of the title
compound is also described in W009105717.
B) 1-Methylcyclopropyl 4-nitrophenyl carbonate
A solution of 1-methylcyclopropanol (10 g, 137 mmol), 4-nitrophehyl
chloroformate (32
g, 152 mmol), and a few crystals of 4-dimethylaminopyridine (150 mg, 1.2 mmol)
in
dichloromethane (462 mL), was cooled to zero degree Celsius. Triethylamine
(36.5 g,
361 mmol) was added drop-wise. After 10 minutes, the ice bath was removed and
the
reaction was allowed to stir at room temperature for 14 hours. The reaction
mixture was
washed twice with saturated aqueous sodium carbonate. The aqueous phase was
extracted with dichloromethane. The combined organic extracts were washed with
water, dried over magnesium sulfate, filtered and the filtrate concentrated in
vacuo. The
residue was purified by flash silica gel chromatography, eluting with a
gradient mixture
of ethyl acetate in heptane (0 to 5% ethyl acetate over the first 10 minutes,
then
isocratic at 5% ethyl acetate to heptane) to give 20.8 g of the desired
carbonate as a
clear oil. This oil solidified upon standing.
1H NMR (500 MHz, deuterochloroform) delta 0.77 (app. t, J=6.59 Hz, 2 H), 1.09
(app. t,
J=7.07 Hz, 2 H), 1.67 (s, 3 H), 7.40 (app. dt, J=9.27, 3.17 Hz, 2 H), 8.29
(app. dt,
J=9.27, 3.17 Hz, 2 H).
Alternatively the 1-methylcyclopropanol can be prepared as follows:
1-Methylcyclopropanol
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A 2000 mL 4-neck flask was equipped with a mechanical stirrer, inert gas
inlet,
thermometer, and two pressure - equalizing addition funnels. The flask was
flushed with
nitrogen and charged with 490 mL of diethyl ether followed by 18.2 mL (30
mmol) of
titanium tetra(2-ethylhexyloxide). One addition funnel was charged with a
solution
prepared from 28.6 mL (360 mmol) of methyl acetate diluted to 120 mL with
ether. The
second addition funnel was charged with 200 mL of 3 M ethylmagnesium bromide
in
ether solution. The reaction flask was cooled in an ice water bath to keep the
internal
temperature at 10 degrees Celsius or below. Forty milliliters of the methyl
acetate
solution was added to the flask. The Grignard reagent was then added drop-wise
from
the addition funnel at a rate of about 2 drops every second, and no faster
than 2 mL per
minute. After the first 40 mL of Grignard reagent had been added, another 20
mL
portion of methyl acetate in ether solution was added. After the second 40 mL
of
Grignard reagent had been added, another 20 mL portion of methyl acetate in
diethyl
ether solution was added. After the third 40 mL of Grignard reagent had been
added,
another 20 mL portion of methyl acetate in ether solution was added. After the
fourth 40
mL of Grignard reagent had been added, the last 20 mL portion of methyl
acetate in
ether solution was added.
The mixture was stirred for an additional 15 minutes following the completion
of the
addition of Grignard reagent. The mixture was then poured into a mixture of
660 g of ice
and 60 mL of concentrated sulfuric acid with rapid stirring to dissolve all
solids. The
phases were separated and the aqueous phase was extracted again with 50 mL of
diethyl ether. The combined ether extracts were washed with 15 mL of 10%
aqueous
sodium carbonate, 15 mL of brine, and dried over 30 grams magnesium sulfate
for 1
hour with stirring. The ether solution was then filtered. Tri-n-butylamine
(14.3 mL, 60
mmol) and mesitylene (10 mL were added. Most of the diethyl ether was removed
by
distillation at atmospheric pressure using a 2.5 cm x 30 cm jacketed Vigreux
column.
The remaining liquid was transferred to a smaller distillation flask using two
10 mL
portions of hexane to facilitate the transfer. Distillation at atmospheric
pressure was
continued through a 2 cm x 20 cm jacketed Vigreux column. The liquid
distilling at 98 -
105 C was collected to provide 14 g of the title compound as a colorless
liquid. 1H
NMR (400 MHz, deuterochloroform) delta 0.42 - 0.48 (m, 2 H), 0.74 - 0.80 (m, 2
H),
1.45 (s, 3 H), 1.86 (br. s., 1 H).
Preparation 27: 2-fluoro-4-(1-methyl-1H-imidazol-5-yl)phenol
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i N,I
HO* NI
F
A) 5-(3-Fluoro-4-methoxyphenyI)-1-methyl-1H I midazole
2-Fluoro-4-bromo anisole (0.216 mL, 1.63 mmol), tri(2-furyl)phosphine (25.9
mg, 0.108
mmol), and potassium carbonate (300 mg, 2.17 mmol) were placed in a microwave
vial
and dissolved in anhydrous N,N-dimethylformamide (4.8 mL). The mixture was
degassed with a stream of nitrogen gas for 10 minutes, 1-methylimidazole
(0.087 mL,
1.1 mmol) and palladium(II) acetate (12.4 mg, 0.054 mmol) were added, and the
mixture was degassed for another 10 minutes. The vessel was placed in a
microwave
reactor at 140 degrees Celsius for 2 hours. The mixture was diluted with ethyl
acetate,
filtered through Celite , and the filtrate was concentrated under reduced
pressure. The
crude material was purified by chromatography eluting with a 25 to 100% ethyl
acetate
in heptane then 0 to10 /0 methanol in dichloromethane gradient to give the
title
compound as a yellow oil (210 mg). 1H NMR (500 MHz, deuterochloroform) delta
3.57
(s, 3 H), 3.85 (s, 3 H), 6.95 - 6.98 (m, 2 H), 7.00 - 7.07 (m, 2 H), 7.42 (s,
1 H). Proton
shift at 7.42 is indicative of desired imidazole isomer as compared to
literature (Eur. J.
Org. chem., 2008, 5436 and Eur. J. Org., 2006, 1379).
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B) 2-Fluoro-4-(1-methyl-1H-imidazol-5-yl)phenol
To a solution of 5-(3-fluoro-4-methoxyphenyI)-1-methyl-1H lmidazole (101.8 mg,
0.494
mmol) in dichloromethane (2.0 mL ) was added a solution of boron(III) bromide
(0.50
mL, 1.0 M solution in heptane) at -30 degrees Celsius. The mixture was stirred
at room
temperature for 20 hours. The mixture was then cooled to -30 degrees Celsius
and
methanol (2 mL) was added to the mixture. The mixture was concentrated in
vacuo, and
the residue was dissolved in water and neutralized with 1M sodium hydroxide.
The
solution was concentrated to give the title compound as a yellow solid (90
mg). This
compound was used further without purification.
Preparation 28: 2-Fluoro-4-(1-methyl-1H-imidazol-2-yl)phenol
N
/ 1
HO* NI
F
A) 2-(3-Fluoro-4-methoxyphenyI)-1-methyl-1H I midazole
2-Fluoro-4-bromoanisole (0.256 mL, 1.93 mmol) and copper(I) iodide (375 mg,
1.93
mmol) were placed in a microwave vial and dissolved in N,N-dimethylformamide
(4.8
mL). The mixture was degassed for 10 minutes with a stream of nitrogen gas, 1-
methylimidazole (0.078 mL, 0.96 mmol) and palladium(II) acetate (11 mg, 0.048
mmol) were added, and the mixture was degassed for another 10 minutes. The
vessel
was placed in a microwave reactor at 140 degrees Celsius for 2 hours. The
mixture was
diluted with ethyl acetate (3 mL), poured into saturated aqueous ammonium
chloride
solution, stirred in the open air for 30 minutes, and extracted twice with
ethyl acetate.
The combined organic phases were washed with water, dried over sodium sulfate,
filtered and the filtrate was concentrated in vacuo. The crude material was
purified by
chromatography, eluting with a gradient mixture of ethyl acetate to heptane
(25 to 100`)/0
ethyl acetate/heptane then 0 to10 /0 methanol in dichloromethane) to give 2-(3-
fluoro-4-
methoxypheny1)-1-methyl-1H lmidazole as a yellow oil (35.8 mg). 1H NMR (400
MHz,
deuterochloroform) delta 3.66 (s, 3 H), 3.88 (s, 3 H), 6.90 (s, 1 H), 6.96 (m
1 H), 7.10 (s,
1 H), 7.24 - 7.33 (m, 2 H). Proton NMR indicates desired imidazole isomer as
compared
to the proton NMR of 5-(3-fluoro-4-methoxyphenyI)-1-methyl-1H Imidazole
(preparation
27) and the literature Eur. J. Org. chem., 2008, 5436 and Eur. J. Org., 2006,
1379).
B) 2-Fluoro-4-(1-methyl-1H-imidazol-2-yl)phenol
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2-Fluoro-4-(1-methyl-1H-imidazol-2-yl)phenol was prepared from 2-(3-fluoro-4-
methoxypheny1)-1-methyl-1H lmidazole following a procedure analogous to that
in
Preparation 27 (B) to give the title compound as a brown solid (33.4 mg). The
crude
material was used further without purification.
Preparation 29: 2-Fluoro-4-(methylsulfonyI)-1-(prop-1-en-2-yl)benzene
F
9'.
S\
' b
To a solution of 1-bromo-2-fluoro-4-(methylsulfonyl)benzene (199 mg, 0.790
mmol) and
potassium isopropenyltrifluoroborate (300 mg, 2.57 mmol) in 2-propanol (10 mL)
was
added the catalyst 1,1'-bis-(diphenylphosphino)-ferrocene palladium dichloride
(67 mg,
0.089 mmol) and triethylamine (0.17 mL, 1.20 mmol) sequentially. The reaction
was
heated at 90 degrees Celsius for 15 hours, and then the reaction was stirred
at room
temperature for 48 hours. Water and ethyl acetate were then added, and the
layers
were separated. The aqueous layer was extracted with ethyl acetate. The
organic
extracts were combined, washed with brine and dried over sodium sulfate. The
mixture
was filtered, and the filtrate concentrated under reduced pressure. The
residue was
purified by silica gel chromatography (10 to 100% ethyl acetate in heptane) to
give the
title compound as a white solid (130 mg, 80%). 1H NMR (500 MHz,
deuterochloroform)
delta 2.17 (s, 3 H), 3.08 (s, 3 H), 5.29 - 5.43 (m, 2 H), 7.51 (t, J=7.56 Hz,
1 H), 7.64 (dd,
J=9.88, 1.59 Hz, 1 H), 7.70 (dd, J=8.05, 1.71 Hz, 1 H).
Preparation 30: 4-Hydroxy-2-methylbenzonitrile
OH
0
INI
Boron trichloride in dichloromethane (61.2 mL, 1M) was added slowly to
dichloromethane (93 mL) and cooled to -78 degrees Celsius. To this was added a
solution of 4-methoxy-2-methylbenzonitrile (3.00 g, 20.4 mmol) and
tetrabutylammonium iodide (7.17 g, 61.2 mmol) in dichloromethane (20 mL). The
reaction mixture was allowed to stir at -78 degrees Celsius for 5 minutes. The
reaction
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mixture was then gradually warmed to room temperature and stirred for 2 hours.
An ice
slurry was slowly added to quench the reaction. The reaction was allowed to
stir for 30
minutes and the layers were separated. The aqueous layer was extracted with
dichloromethane (2x) and the combined organic extracts were passed through a
phase
separated cartridge and concentrated in vacuo. The crude mixture was purified
by flash
chromatography eluting with 0% to 60% ethyl acetate in pentane to give the
target
compound as a yellow solid (1.85 g, 68 %). 1H NMR deuteromethanol delta ppm
7.40
(d, 1H), 6.80 (s, 1H), 6.70 (d, 1H), 2.40 (s, 3H); GCMS (Cl method) ES+= 133
[M] AP+
= 133 [M].
Preparation 31A: 3-Fluoro-4-hydroxy-N-methylbenzamide
OH
0 F
0 NH
I
A) Benzyl 4-(benzyloxy)-3-fluorobenzoate
3-Fluoro-4-hydroxybenzoic acid (5.00 g, 32.06 mmol), benzyl bromide (8.22 mL,
67.3
mmol) and potassium carbonate (13.3 g, 96.24 mmol) were combined in acetone
and
heated at reflux for 18 hours. The solution was cooled down to room
temperature, the
solids were filtered and the filtrate was diluted with ethyl acetate. The
organic phase
was washed with saturated aqueous brine solution, dried over magnesium
sulfate,
filtered and the filtrate was concentrated under reduced pressure to give the
desired
product benzyl 4-(benzyloxy)-3-fluorobenzoate. 1H NMR (500 MHz,
deuterochloroform)
delta ppm 5.22 (s, 2 H) 5.36 (s, 2 H) 7.03 (t, J=8.42 Hz, 1 H) 7.29- 7.52 (m,
10 H) 7.76 -
7.89 (m, 2 H); LCMS (ES+)= 381.2 (M+45)
B) 4-(Benzyloxy)-3-fluorobenzoic acid
Benzyl 4-(benzyloxy)-3-fluorobenzoate (11.6 g, 34.2 mmol) was dissolved in
tetrahydrofuran (50 mL) and methanol (50 mL). Aqueous sodium hydroxide (70 mL,
1M) was added to the reaction mixture and stirred for 18 hours. The reaction
was
cooled in an ice bath and acidified to pH 3 by careful addition of aqueous 1M
solution of
hydrochloric acid. A white solid precipitated out and was filtered and dried
over night to
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give the desired product, 4-(benzyloxy)-3-fluorobenzoic acid, as a white solid
(7.6 g,
90%). 1H NMR (500 MHz, deuterodimethylsulfoxide) delta ppm 3.32 (br. s., 1 H)
5.27
(s, 2 H) 7.34 - 7.39 (m, 2 H) 7.42 (t, J=7.44 Hz, 2 H) 7.45 - 7.50 (m, 2 H)
7.68 (dd,
J=11.83, 2.07 Hz, 1 H) 7.75 (d, J=8.78 Hz, 1 H)
C) 4-(Benzyloxy)-3-fluoro-N-methylbenzamide
Thionyl chloride (2.7 mL, 37 mmol) was added to a solution of 4-(benzyloxy)-3-
fluorobenzoic acid in dimethylformamide (0.048 mL, 0.617 mmol) and
dichloromethane
(100 mL) at 0 degree Celsius and the resulting solution was stirred at room
temperature
for 20 hours. The reaction was concentrated under reduced pressure and dried
under
high vacuum for 2 hours. The resulting yellow solid was dissolved in
tetrahydrofuran
(60 mL) and a 2M solution of methylamine in tetrahydrofuran (35mL) was added
and the
reaction stirred at room temperature for 18 hours. The reaction mixture was
concentrated under reduced pressure to half the original volume and a white
solid
precipitated out. The solid was filtered off, washed with water and dried in a
vacuum
oven overnight to give the desired product as a white solid (6.00 g, 70%). 1H
NMR (500
MHz, deuterodimethylsulfoxide) delta ppm 2.76 (d, J=4.39 Hz, 3 H) 5.24 (s, 2
H) 7.30 -
7.38 (m, 2 H) 7.41 (t, J=7.32 Hz, 2 H) 7.45 - 7.50 (m, 2 H) 7.58 - 7.75 (m, 2
H) 8.37 (d,
J=4.39 Hz, 1 H)
D) 3-Fluoro-4-hydroxy-N-methylbenzamide
4-(Benzyloxy)-3-fluoro-N-methylbenzamide (1.03 g, 3.97 mmol) was suspended in
ethanol (20 mL) in a Parr bottle. 10% Palladium on carbon (80 mg) in about 1.5
mL of
water was added under a steady stream of nitrogen. The reaction was shaken
under a
50 psi atmosphere of hydrogen at room temperature for 64 hours. The reaction
mixture
was carefully filtered through a pad of Celite washing with copious amounts
of ethyl
acetate. The filtrate was concentrated under reduced pressure to give the
desired
product (628 mg, 93%) as a light brownish yellow solid. 1H NMR (500 MHz,
deuterochloroform) delta ppm 3.02 (d, J=4.88 Hz, 3 H) 7.05 (t, J=8.42 Hz, 1 H)
7.44 (d,
J=9.76 Hz, 1 H) 7.60 (dd, J=11.10, 2.07 Hz, 1 H)
Preparation 31B: 3-Fluoro-4-hydroxy-N,N-dimethylbenzamide
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OH
0 F
0 N
1
3-Fluoro-4-hydroxybenzoic acid (2.00 g, 12.8 mmol), dimethylamine
hydrochloride (4.28
g, 20.5 mmol), 1-hydroxy benzotriazole monohydrate (1.96 g, 12.8 mmol), and
diisopropylethyl amine (4.5 mL, 26 mmol) were combined in dichloromethane. 1-
Ethyl-
3-(3-dimethylaminopropyl) carbodiimide hydrochloride (3.93 g, 20.5 mmol) was
added,
and the reaction vessel was flushed with nitrogen, capped, and stirred
overnight at room
temperature. The reaction was diluted with dichloromethane and 1M phosphoric
acid.
The precipitate that formed was filtered off and the dichloromethane layer was
washed
with dilute aqueous sodium bicarbonate and brine, dried over sodium sulfate,
filtered,
and the filtrate was concentrated under reduced pressure. The residue was
purified by
flash chromatography eluting with 65% ethyl acetate in heptanes to give the
desired
product (218 mg, 9%). LC/MS (ES+): 184.1 (M+1)
Preparation 32: tert-Butyl 3-Hydroxy-4,4-dimethoxypiperidine-1-carboxylate
HO
N¨ (µ
tert-Butyl 4-oxo-1-piperidinecarboxylate (2.00 g, 10 mmol) was dissolved in
methanol
(20 mL) and cooled to 0 degrees Celsius. Powdered potassium hydroxide (1.26 g,
22.1
mmol) was added. Iodine (2.8 g, 11 mmol) was dissolved in methanol (25 mL) and
was
added drop wise to the reaction over 45 minutes. The reaction was then slowly
warmed
up to room temperature and stirred for 16 hours. The reaction was concentrated
and
toluene (50 mL) was added. The resulting solids were filtered off and washed
with
toluene. The filtrate was concentrated under reduced pressure and the residue
was
purified by flash chromatography eluting with a gradient from 30% to 100%
ethyl acetate
in heptane to give tert-butyl 3-hydroxy-4,4-dimethoxypiperidine-1-carboxylate
(1.89 g,
72%). 1H NMR (deuteromethanol, 400 MHz) delta ppm 4.06-4.00 (m, 1H), 3.99-3.91
(m,
1H), 3.80-3.73(m, 1H), 3.29(s, 3H), 3.28(s, 3H), 3.22-3.10 (br m, 1H), 2.95-
2.80 (br m,
1H), 1.91-1.77 (m, 2H), 1.50 (s, 9 H).
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Preparation 33: tert-Butyl 3-Hydroxy-4-oxopiperridine-1-carboxylate
HO
0 (
0 )--\N-µ
0
tert-Butyl 3-hydroxy-4,4-dimethoxypipendine-1-carboxylate (6.70g, 26 mmol) was
dissolved in acetone (135 mL), and p-toluene sulfonic acid (244 mg, 1.28 mmol)
was
added. The reaction was stirred at room temperature for 16 hours. The mixture
was
concentrated and the resulting residue was dissolved in tert-butyl methyl
ether and
washed with saturated aqueous sodium bicarbonate solution. The organic layer
was
dried over sodium sulfate, filtered and the filtrate was concentrated under
reduced
pressure to give tert-butyl 3-hydroxy-4-oxopiperridine-1-carboxylate as an oil
(4.67 g,
69%). GC/MS (method 1): R, = 4.95 min; MS (ESIpos): m/z = 159 [M-tBu].
Preparation 34: tert-Butyl 4-Hydrazino-3-hydroxypipendine-1-carboxylate
HO
H2N, -\ 0 K
HN_N-µ
/ 0
tert-Butyl 3-hydroxy-4-oxopiperridine-1-carboxylate (5.50 g, 26 mmol) was
dissolved in
methanol (120 mL) and degassed with a stream of nitrogen in a capped Parr
Shaker
bottle. Hydrazine-hydrochloride (1.44 mg, 21 mmol) was dissolved in water (20
mL) and
added to the reaction. The flask was rinsed with 5 mL of water and it was also
added to
the reaction. 10% Platinum on carbon catalyst (500 mg) was slurried in water
and
added to the reaction mixture. The mixture was shaken under at 50 psi
atmosphere of
hydrogen at room temperature for 16 hours. The reaction was filtered through a
pad of
Celite washing with methanol. The filtrate was concentrated under reduced
pressure
and then diluted with heptanes and concentrated under reduced pressure to give
the
desired product, tert-butyl 4-hydrazino-3-hydroxypipendine-1-carboxylate.
Preparation 35: tert-Butyl 4-1-5-Amino-4-(ethoxycarbony1)-1H-pyrazol-1-y11-3-
hydroxypipendine-1-carboxylate
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HO
.....N, \ 0 K
7-µ0
0 NH2
1
tert-Butyl 4-hydrazino-3-hydroxypiperidine-1-carboxylate (5.30 g, 20 mmol) and
ethyl(ethoxymethlene)cyanoacetate (3.42 g, 19.8 mmol) were combined in
absolute
ethanol (170 mL). Sodium acetate trihydrate (10.90 g, 79.2 mmol) was added,
and the
reaction mixture was heated at reflux for 4 hours. The reaction was cooled to
room
temperature, concentrated under reduced pressure and the resulting residue was
diluted with water and ethyl acetate. The organic layer was dried over sodium
sulfate,
filtered and the filtrate was concentrated under reduced pressure. The crude
oil was
purified by flash chromatography eluting with a gradient from 10% to 100%
ethyl acetate
in heptanes to give tert-butyl 4-[5-amino-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-
3-
hydroxypiperidine-1-carboxylate.
Preparation 36: tert-Butyl 4-[5-amino-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-
fluoropiperidine-1-carboxylate
F
NH2
/0
1
tert-Butyl 4-[5-amino-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-hydroxypiperidine-
1-
carboxylate (1.71 g, 4.82 mmol) was dissolved in dichloromethane (50 mL) and
cooled
to -78 degrees Celsius. Diethylaminosulfur trifluoride (0.710 mL, 0.58 mmol)
was added
drop wise, and then warmed up to 0 degrees Celsius for 25 minutes. The
reaction
solution was cooled to -78 degrees Celsius and methanol (10 mL) carefully
added. The
reaction was concentrated under reduced pressure and the residue was purified
by
flash chromatography eluting with a gradient from 10% to 100% ethyl acetate in
heptanes to give tert-butyl 4-[5-amino-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-
fluoropiperidine-1-carboxylate.
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Preparation 37: tert-Butyl 415-bromo-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-
fluoropiperidine-1-carboxylate
F
NI, t 0 K
ON 71¨µ0
0 Br
1
tert-Butyl 445-amino-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-fluoropiperidine-1-
carboxylate (710 mg, 1.99 mmol) was dissolved in acetonitrile (25 mL). Copper
(II)
bromide (539 mg, 2.39 mmol) was added and the reaction was heated to 60
degrees
Celsius. tert-Butyl nitrile (0.315 mL, 2.9 mmol) was added drop wise and the
mixture
was heated at 65 degrees Celsius for 15 minutes. The reaction was cooled to
room
temperature and poured into cold 1N hydrochloric acid and extracted with ethyl
acetate
(2x). The combined organic extracts were washed with saturated aqueous sodium
bicarbonate and brine and dried over sodium sulfate, filtered and the filtrate
was
concentrated under reduced pressure. The crude residue was purified by flash
chromatography eluting with a gradient from 10% to 50% ethyl acetate in
heptanes to
give tert-butyl 445-bromo-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-
fluoropiperidine-1-
carboxylate (320 mg, 38%). 1H NMR (500 MHz, deuterochloroform) delta ppm 1.37
(t,
3 H) 1.49 (s, 9 H) 1.98 (d, J=13.42 Hz, 1 H) 2.12 -2.26 (m, 1 H) 2.90 (br. s.,
2 H) 4.18
(br. s., 1 H) 4.33 (q, J=7.24 Hz, 2 H) 4.44 - 4.70 (m, 2 H) 4.85 - 5.05 (m, 1
H) 8.04 (s, 1
H)
Preparation 38: Ethyl 5-cyano-1-(3-fluoropiperidin-4-yI)-1H-pyrazole-4-
carboxylate
F
ji-N..N --- \N H
0 --- ___ /
0 \\
( N
tert-Butyl 445-bromo-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-fluoropiperidine-1-
carboxylate (185 mg, 0.31 mmol), 1,1'bis (diphenylphosphino)ferrocene (18 mg,
0.032
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mmol), zinc dust (18 mg, 0.27 mmol), zinc cyanide (39.1 mg, 0.33 mmol) and 10%
palladium black (19.2 mg, 0.021 mmol) were combined in dimethylacetamide (3
mL) in
a microwave vial. The reaction mixture was degassed with nitrogen and heated
at 170
degrees Celsius for 4.5 hours. The reaction mixture was cooled to room
temperature
and diluted with ethyl acetate. The reaction was filtered through a pad of
Celite and
the filtrate was diluted with water and extracted with ethyl acetate (2x). The
combined
organic extracts were washed with water then brine and dried over sodium
sulfate,
filtered and the filtrate was concentrated under reduced pressure. The crude
residue
was purified by flash chromatography eluting with a gradient from 10% to 100%
ethyl
acetate in heptanes to give ethyl 5-cyano-1-(3-fluoropiperidin-4-yI)-1H-
pyrazole-4-
carboxylate (80 mg, 98%).
Preparation 39: tert-Butyl 4-[5-cyano-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-
fluoropiperidine-1-carboxylate
F
;............NN___¨\_µ0 (
1 N
Ethyl 5-cyano-1-(3-fluoropiperidin-4-yI)-1H-pyrazole-4-carboxylate (60 mg,
0.22 mmol)
was dissolved in tetrahydrofuran (3 mL) and triethylamine (40 uL, 0.27 mmol)
was
added. Di-tert-butyl dicarbonate (50 mg, 0.225 mmol) was added and the
reaction was
stirred at room temperature under nitrogen for 3 hours. The reaction was
concentrated
under reduced pressure and the residue was purified by flash chromatography
eluting
with a gradient from 10% to 100% ethyl acetate in heptanes to give tert-butyl
445-
cyano-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-fluoropiperidine-1-carboxylate as
an oil (52
mg, 63%).
Preparation 40: 141-(tert-Butoxycarbony1)-3-fluoropiperidin-4-y1]-5-cyano-1H-
pyrazole-
4-carboxylic acid
OH
/
0
N \ N¨µ
N 0
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tert-Butyl 445-cyano-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-fluoropiperidine-1-
carboxylate (80 mg, 0.22 mmol) was dissolved in tetrahydrofuran (2.5 mL),
water (1.5
mL) and methanol (0.4 mL). The solution was cooled to 0 degrees Celsius and
lithium
hydroxide monohydrate (19 mg, 0.436 mmol) was added. The reaction was slowly
allowed to warm up to room temperature over 2.5 hours. The reaction mixture
was
concentrated; the residue was dissolved in water and extracted with ethyl
acetate and
methyl tert-butyl ether. The organic layer was extracted with water. The
combined
aqueous extracts were acidified with 1N aqueous sodium bisulfate to pH 2. The
acidic
solution was extracted with ethyl acetate (3x) and the extracts were washed
with brine,
dried over sodium sulfate, filtered and the filtrate was concentrated under
reduced
pressure to give 141-(tert-butoxycarbony1)-3-fluoropiperidin-4-y1]-5-cyano-1H-
pyrazole-
4-carboxylic acid as a white solid.
Preparation 41: tert-Butyl 4-15-cyano-4-(hydroxymethyl)-1H-pyrazol-1-y11-3-
fluoropiperidine-1-carboxylate
F
.......N N--µ
0
OH \\\
N
Freshly recrystallized (from heptanes) cyanuric chloride (78 mg, 0.414 mmol)
was
dissolved in dimethoxyethane (2 mL) and 4-methyl-morpholine (0.020 mL, 0.215
mmol)
was added. To this gummy solution was added 141-(tert-butoxycarbony1)-3-
fluoropiperidin-4-y1]-5-cyano-1H-pyrazole-4-carboxylic acid (70 mg, 0.21 mmol)
dissolved in dimethoxyethane (2 mL). The reaction was heated at 60 degrees
Celsius
for 3 hours. The reaction was cooled to room temperature and filtered through
a pad of
Celite washing with dimethoxyethane. The filtrate was cooled to 0 degrees
Celsius
and a solution of sodium borohydride (17 mg, 0.474 mmol) dissolved in water
(0.4 mL)
was added very slowly (drop wise). Once addition was complete, the reaction
was
allowed to warm up to room temperature for 2.5 hours. The reaction solution
was
further diluted with water and acidified to pH 2.5 using 1M sodium bisulfate.
The
aqueous layer was extracted with ethyl acetate (2x) and the combined organic
layers
were dried over sodium sulfate, filtered and the filtrate was concentrated
under reduced
pressure. The crude residue was purified by flash chromatography eluting with
a
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gradient from 10% to 100% ethyl actetate in heptanes to give tert-butyl 445-
cyano-4-
(hydroxymethyl)-1H-pyrazol-1-y1]-3-fluoropiperidine-1-carboxylate as an oil
(28 mg,
42%).
Preparation 42: tert-Butyl 4-(5-cyano-4-{[(methylsulfonyl)oxylmethyll-1H-
pyrazol-1-y1)-
3-fluoropiperidine-1-carboxylate
t\ 0 (
N N-µ
____________________________________________ 0
01 6N
tert-Butyl 445-cyano-4-(hydroxymethyl)-1H-pyrazol-1-y1]-3-fluoropiperidine-1-
carboxylate (28 mg, 0.086 mmol) was dissolved in dichloromethane (3 mL).
Triethylamine (0.036 mL, 0.258 mmol) was added and the mixture was cooled to 0
degrees Celsius. Methanesulfonic anhydride (20 mg, 0.112 mmol) was added drop
wise and slowly allowed to warm up to room temperature over 2 hours.
Dichloromethane and saturated aqueous sodium bicarbonate were added to the
reaction solution and the biphasic solution was separated. The aqueous layer
was
extracted with dichloromethane (2x) and the combined organic extracts were
passed
through a plug of cotton. The filtrate was concentrated under reduced pressure
to give
tert-butyl 4-(5-cyano-4-{[(methylsulfonyl)oxy]methyll-1H-pyrazol-1-y1)-3-
fluoropiperidine-
1-carboxylate as an oil (33 mg, 95%). 1H NMR (500 MHz, deuterochloroform)
delta
ppm 1.25 - 1.30 (m, 2 H) 1.50 (s, 9 H) 2.01 - 2.06 (m, 1 H) 2.75 - 2.85 (m, 2
H) 3.08 (s,
3 H) 4.64 - 4.74 (m, 1 H) 4.79 - 4.98 (m, 2 H) 5.26 (s, 2 H) 7.75 (s, 1 H)
Preparation 43: Isomers of tert-Butyl-3-fluoro-4-hydroxypiperidine-1-
carboxylate (B and
The experimental details are described in detail in Scheme 4 below.
Scheme 4
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0 0
Step A Step B
00< 00< 00<
Step C
V
chiral
OH OH OH racemic OH
Step D
00<
0 0 00< 0 0
Step A) tert-Butyl-4-[(trimethylsilypoxyl-3,6-dihydropyridine-1(2H)-
carboxylate
00<
To a solution of N-tert-butoxycarbony1-4-piperidone (30.0 g, 0.15 mol) in dry
N,N-
dimethylformamide (300 mL) at room temperature was added trimethylsilyl
chloride
(22.9 mL, 0.18 mol) and triethylamine (50.4 mL, 0.36 mol) successively via
addition
funnels. The resulting solution was heated at 80 degrees Celsius overnight and
then
cooled to room temperature. The reaction mixture was diluted with water and
heptane.
The layers were separated, and the aqueous layer was extracted with heptane.
The
combined heptane layers were washed sequentially with water and brine and then
dried
over magnesium sulfate. The mixture was filtered, and the filtrate
concentrated under
reduced pressure to give the crude product as a yellow oil. The oil was
purified by
passing it through a plug of silica gel eluting with 9:1 heptane/ethyl acetate
to give the
title compound as a colorless oil (33.6 g, 82%). 1H NMR (400 MHz,
deuterochloroform)
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delta 4.78 (br s, 1H), 3.86 (br s, 2H), 3.51 (t, 2H), 2.09 (br s, 2H), 1.45
(s, 9H), 0.18 (s,
9H).
Step B) tert-Butyl-3-fluoro-4-oxooioeridine-1-carboxylate
ci
)F
---
N
0 0
To a stirred solution of tert-butyl-4-[(trimethylsilypoxy]-3,6-dihydropyridine-
1(2H)-
carboxylate (28.8 g, 0.11 mol) in acetonitrile (300 mL) at room temperature
was added
SelectfluorTM (41.4 g, 0.12 mol). The resulting pale yellow suspension was
stirred at
room temperature for 1.5 hours. Saturated aqueous sodium bicarbonate (300 mL)
and
ethyl acetate (300 mL) were added, and the layers were separated. The aqueous
layer
was extracted twice with ethyl acetate, and all the organic layers were
combined and
washed sequentially with saturated aqueous sodium bicarbonate and brine and
then
dried over magnesium sulfate. The mixture was filtered, and the filtrate was
concentrated under reduced pressure to give the crude product as a pale yellow
oil.
Purification of this material by repeated column chromatography on silica gel
with
heptane/ethyl acetate gradient (2:1 to 1:1) gave the title compound as a white
solid
(15.5 g, 67%). 1H NMR (400 MHz, deuterochloroform): delta 4.88 (dd, 0.5 H),
4.77 (dd,
0.5H), 4.47 (br s, 1H), 4.17 (ddd, 1H), 3.25 (br s, 1H), 3.23 (ddd, 1H), 2.58
(m, 1H), 2.51
(m, 1H), 1.49 (s, 9H).
Alternatively Step B can be performed as follows, isolating the hydrate of the
ketone.
To a stirred solution of tert-butyl-4-[(trimethylsilypoxy]-3,6-dihydropyridine-
1(2H)-
carboxylate (41.3 g, 0.15 mol) in acetonitrile (500 mL) at room temperature
was added
SelectfluorTM (56.9 g, 0.16 mol). The resulting pale yellow suspension was
stirred at
room temperature for 4 hours 10 minutes. Saturated aqueous sodium bicarbonate
and
ethyl acetate were added, and the layers were separated. The aqueous layer was
extracted twice with ethyl acetate, and all the organic layers were combined
and
washed sequentially with saturated aqueous sodium bicarbonate and brine and
then
dried over magnesium sulfate. The mixture was filtered, and the filtrate was
concentrated under reduced pressure to give the crude tert-butyl-3-fluoro-4-
oxopiperidine-1-carboxylate as white solid. The crude tert-butyl-3-fluoro-4-
oxopiperidine-1-carboxylate was suspended in tetrahydrofuran (120 mL) and
water (120
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mL) was added. The resulting solution was stirred at room temperature for 5.5
hours
and then concentrated under reduced pressure. The residue was dried under high
vacuum, transferred to an Erlenmeyer flask, and suspended in dichloromethane
(250
mL). The resulting suspension was stirred for 5 minutes and the solids
collected by
filtration using a sintered glass funnel. The resulting filter cake was
thoroughly washed
with dichloromethane (200 mL), a 1:1 mixture of dichloromethane (200 mL) and
heptane
(100 mL). The solid was then dried under high vacuum to provide tert-butyl 3-
fluoro-4,4-
dihydroxypiperidine-1-carboxylate (26.4 g). 1H NMR (500 MHz, deutero dimethyl
sulfoxide) delta 1.38 (s, 9 H), 1.49-1.52 (m, 1H), 1.63-1.68 (m, 1 H), 2.82 -
3.20 (m, 2 H)
3.75 (br, 1 H), 3.97 (br, 1 H), 4.12 (d, J = 45, 1 H), 5.92 (s, 1 H), 5.97 (s,
1 H).
Step C) Isomers of (R*)-tert-Buty1-3-(S)-fluoro-4-(R)-hydroxypiperidine-1-
carboxylate
(racemic)
OH OH
N --...
N
00< 00<
To a solution of tert-butyl-3-fluoro-4-oxopiperidine-1-carboxylate (15.5 g,
71.3
mmol) in methanol (150 mL) at 0 degrees Celsius was added sodium borohydride
(3.51
g, 93.7 mmol). The resulting mixture was stirred at 0 degrees Celsius for 2
hours and
then allowed to warm to room temperature. Saturated aqueous ammonium chloride
(200 mL) was added, and the mixture was extracted three times with ethyl
acetate. The
combined extracts were washed with brine and dried over magnesium sulfate. The
mixture was filtered, and the filtrate was concentrated under reduced pressure
to give
the crude product mixture which was purified by column chromatography on
silica gel
eluting with heptane-ethyl acetate (3:2 - 1:1) to give the first eluting
product, tert-butyl-
(3,4-trans)-3-fluoro-4-hydroxypiperidine-1-carboxylate (compound C, Scheme 4)
(3.81
g, 24%), as a pale yellow oil which solidified on standing to a white solid.
1H NMR (400
MHz, deuterochloroform) delta 4.35 (ddd, 0.5 H), 4.18 (ddd, 0.5 H), 4.15 (br
s, 1H),
3.89-3.74 (m, 2H), 2.97 (br s, 1H), 2.93 (ddd, 1H), 2.47 (s, 1H), 2.05-1.92
(m, 1H), 1.58-
1.46 (m, 1H), 1.44 (s, 9H).
The second eluting compound, tert-butyl-(3,4-cis)-3-fluoro-4-hydroxy-
piperidine-
1-carboxylate (compound B, Scheme 4) (10.57 g, 68%) was then isolated as a
white
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solid. 1H NMR (400 MHz, deuterochloroform) delta 4.69 - 4.65 (m, 0.5H), 4.53-
4.49 (m,
0.5H), 3.92 - 3.86 (m, 2H), 3.69 (br s, 1H), 3.39 (br s, 1H), 3.16 (br s, 1H),
2.13 (s, 1H),
1.88 - 1.73 (m, 2H), 1.44 (s, 9H).
Alternatively Step C can be performed starting with the hydrate tert-butyl 3-
fluoro-
4,4-dihydroxypiperidine-1-carboxylate (Step 2) as follows.
To a stirred solution of tert-butyl 3-fluoro-4,4-dihydroxypiperidine-1-
carboxylate
(20.0 g, 85 mmol) in tetrahydrofuran (500 mL) at -35 degrees Celsius was added
a
solution of L-Selectride in tetrahydrofuran (170 mL, 1 M, 170 mmol) drop-wise
over 30
minutes. The reaction mixture was warmed to 0 degree Celsius over 1.5 h. The
reaction
mixture was quenched with saturated aqueous ammonium chloride (150 mL) and
vigorously stirred for 15 minutes. To this 0 degree Celsius mixture was added
pH 7
phosphate buffer (150 mL), followed by drop-wise addition of a 35% aqueous
hydrogen
peroxide solution (150 mL). The resulting mixture was stirred for 30 minutes
and diluted
with ethyl acetate. The organic layer was separated and sequentially with
water,
saturated aqueous sodium thiosulfate and brine. The organic layer was then
dried over
anhydrous magnesium sulfate, filtered and the filtrate was concentrated under
reduced
pressure give the crude product mixture which was purified by column
chromatography
on silica gel [ combiflash ISCO 330 g column] eluting with heptane-ethyl
acetate (10 to
60% gradient) to give tert-butyl-(3,4-cis)-3-fluoro-4-hydroxypiperidine-1-
carboxylate
(13.9g).
Step D) Enantiomers of tert-butyl-(3,4-cis)-3-fluoro-4-hydroxy-piperidine-1-
carboxylate
A 1 gram sample of racemic tert-butyl-(3,4-cis)-3-fluoro-4-hydroxy-piperidine-
1-
carboxylate was purified into its enantiomers via preparatory high pressure
liquid
chromatography utilizing a Chiralpak AD-H column (10 x 250 mm) with a mobile
phase
of 90:10 carbon dioxide and ethanol respectively at a flow rate of 10
mL/minute. The
wavelength for monitoring the separation was 210 nM. The analytical purity of
each
enantiomer was determined using analytical high pressure chromatography using
a
Chiralpak AD-H (4.6 mm x 25 cm) column with an isocratic mobile phase of 90:10
carbon dioxide and ethanol respectively at a flow rate of 2.5 mL/minute. The
wavelength
for monitoring the peaks was 210 nm. The following two isomers were obtained:
Compound E, Scheme 4) (3S,4R)-tert-Butyl 3-fluoro-4-hydroxypiperidine-1-
carboxylate,
enantiomer 1 (363 mg): Rt = 2.67 min (100% ee) (optical rotation in
dichloromethane =
+21.2 degrees) and
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OH
LF
..--
N
00<
Compound D, Scheme 4) (3R,4S)-tert-Butyl 3-fluoro-4-hydroxypiperidine-1-
carboxylate,
enantiomer 2 (403 mg): Rt = 2.99 min (88% ee).
OH
---
N
00<
The absolute stereochemistry of the tert-butyl-(3,4-cis)-3-fluoro-4-hydroxy-
piperidine-1-
carboxylate isomers was determined by making a (1S)-(+)-camphorsulfonic acid
salt of
5-(6-((3S,4R)-3-fluoropiperidin-4-yloxy)-5-methylpyrimidin-4-yI)-1-methyl-
1,4,5,6-
tetrahydropyrrolo[3,4-c]pyrazole (see by analogy the preparation in racemic
form
below), prepared using enanantiomer 1 above.
NN NH
rcry),:)," \F ): HO3S 0
1 \
N'N
1
Preparation of 5-(6-{[(3,4-cis)-3-fluoropiperidin-4-yl]oxy}-5-methylpyrimidin-
4-y1)-1-
methyl-1,4,5,6-tetrahydropyrrolo[3,4-Plpyrazole (racemic)
A. Preparation of 5-(6-Chloro-5-methylpyrimidin-4-yI)-1-methyl-1,4,5,6-
tetrahydropyrrolo[3,4-dpyrazole
N N
Ir___91C1
\
N._ N
1
1-Methyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole bis-hydrochloride salt (2.00
g,
10.2 mmol) and 4,6-dichloro-5-methylpyrimidine (1.66 g, 10.2 mmol) were
suspended in
tetrahydrofuran (51 mL) at room temperature. To this was added triethylamine
(4.41
mL, 31.6 mmol), which caused cloudiness in the mixture and led to a brown
solid
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sticking to the flask walls. This mixture was stirred at room temperature for
4 hours and
then heated 50 degrees Celsius for an additional 19 hours. The reaction
mixture was
cooled to room temperature and diluted with water (100 mL). This mixture was
extracted with ethyl acetate (3 x 100 mL). The organic extracts were pooled,
washed
with brine, dried over sodium sulfate, and filtered. The filtrate was reduced
to dryness
under vacuum to yield the title compound as a light brown solid (1.95 g, 78%),
which
was used in the next step without further purification.
1H NMR (500 MHz, deuterochloroform) delta 2.54 (s, 3 H) 3.88 (s, 3 H) 4.90
(app. d,
J=3.66 Hz, 4 H) 7.28 (s, 1 H) 8.29 (s, 1 H).
B. Preparation of tert-Butyl (3,4-cis)-3-fluoro-4-{1-5-methyl-6-(1-methyl-4,6-
dihydropyrrolo[3,4-clpyrazol-5(1H )-yppyrimidin-4-ylloxylpiperidine-1-
carboxylate
(racemic)
0
N N NAO<
[1 ---J
N , N
1
A mixture of tert-butyl (3,4-cis)-3-fluoro-4-hydroxypiperidine-1-carboxylate
(1.67 g, 7.62
mmol) and 5-(6-chloro-5-methylpyrimidin-4-yI)-1-methyl-1,4,5,6-
tetrahydropyrrolo[3,4-
c]pyrazole prepared above (900 mg, 3.60 mmol) was dissolved in 1,4-dioxane (20
mL)
and was heated to 105 degrees Celsius. After heating for 10 minutes, all the
materials
had gone into solution, and sodium bis(trimethylsilyl)amide (4.3 mL, 4.3 mmol,
1M in
toluene) was rapidly added to the mixture, resulting in a cloudy yellow
mixture that was
then stirred for 2 hours at 105 degrees Celsius. The reaction was then cooled
to room
temperature and quenched by adding an equal volume mixture of water and
saturated
aqueous sodium bicarbonate solution. The mixture was extracted with ethyl
acetate (3
x 15 mL). The combined organic extracts were washed with brine, dried over
sodium
sulfate, and filtered. The filtrate was concentrated under vacuum to give a
yellow
residue that was purified by column chromatography on silica gel eluting with
60 to
100% ethyl acetate in heptane. A mixture of the title compound and the
starting 5-(6-
chloro-5-methylpyrimidin-4-y1)-1-methyl-1,4,5,6-tetrahydropyrrolo[3,4-
c]pyrazole was
isolated as a white solid (1.20 g) and was used without further purification
in subsequent
reactions.
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A batch of crude tert-butyl (3,4-cis)-3-fluoro-4-{[5-methyl-6-(1-methyl-4,6-
dihydropyrrolo[3,4-c]pyrazol-5(1H )-yppyrimidin-4-yl]oxylpiperidine-1-
carboxylate from a
separate reaction, run under the same conditions, was purified by HPLC. The
crude
sample (9.5 mg) was dissolved in dimethyl sulfoxide (1 mL) and purified by
preparative
reverse phase HPLC on a Waters XBridge C18 19 x 100 mm, 0.005 mm column,
eluting
with a linear gradient of 80% water/acetonitrile (0.03% ammonium hydroxide
modifier)
to 0% water/acetonitrile in 8.5 minutes, followed by a 1.5 minute period at 0%
water/acetonitrile; flow rate: 25mL/minute. The title compound (5 mg) was thus
obtained. Analytical LCMS: retention time 2.81 minutes (Waters XBridge C18 4.6
x 50
mm, 0.005 mm column; 90% water/acetonitrile linear gradient to 5%
water/acetonitrile
over 4.0 minutes, followed by a 1 minute period at 5% water/acetonitrile;
0.03%
ammonium hydroxide modifier; flow rate: 2.0 mL/minute); LCMS (ES+) 433.2
(M+1).
C. Preparation of 5-(6-{[(3,4-cis)-3-fluoropiperidin-4-ylloxy}-5-
methylpyrimidin-4-
y1)-1-methyl-1,4,5,6-tetrahydropyrrolor3,4-clpyrazole (racemic)
NN NH
1 ss, j
r_r_111- '0
I \ f
N.N
1
Crude tert-butyl (3,4-cis)-3-fluoro-4-{[5-methyl-6-(1-methyl-4,6-
dihydropyrrolo[3,4-
c]pyrazol-5(1H )-yppyrimidin-4-yl]oxylpiperidine-1-carboxylate (1.20 g)
prepared above
was dissolved in dichloromethane (12 mL) and to this solution was added
trifluoroacetic
acid (5 mL). The reaction was stirred at room temperature for 1 hour. The
solvent was
removed under vacuum, and the residue was dissolved in water (50 mL) and 1N
aqueous hydrochloric acid solution (10 mL). The mixture was extracted with
dichloromethane (10 x 30 mL). The aqueous layer was then brought to pH 12 by
the
addition of 1N aqueous sodium hydroxide solution (20 mL) and was extracted
three
times with dichloromethane (40 mL). The combined organic extracts were washed
with
brine, dried over sodium sulfate and filtered. The filtrate was concentrated
under
reduced pressure to afford 5-(6-{[(3,4-cis)-3-fluoropiperidin-4-yl]oxy}-5-
methylpyrimidin-
4-y1)-1-methyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole (0.72 g, 60% over two
steps) as
a white solid that was used without additional purification.
1H NMR (500 MHz, deuterochloroform) delta 1.84 - 2.08 (m, 2 H) 2.33 (s, 3 H)
2.69 - 2.84 (m, 1 H) 2.83 - 3.01 (m, 1 H) 3.16 (d, J=13.66 Hz, 1 H) 3.27 -
3.44 (m, 1 H)
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3.86 (s, 3 H) 4.78-4.91 (m, 1 H) 4.86 (d, J=1.95 Hz, 2 H) 4.88 (d, J=1.95 Hz,
2 H) 5.21 -
5.32 (m, 1 H) 7.26 (s, 1 H) 8.18 (s, 1 H); LCMS (ES+) 333.4 (M+1).
Please note: Example number begins at 11.
Example 11: Isopropyl 4-{5-cyano-4-[(2,4-difluorophenoxy)methyI]-1H-pyrazol-1-
yllpiperidine-1-carboxylate
F
410 F
CN
0"-N------- __< _____________________________ /\ C)
, -(
N N-µ
0
Isopropyl 4-(5-cyano-4-((methylsulfonyloxy)methyl)-1H-pyrazol-1-y1)piperidine-
1-
carboxylate (Preparation 10) (166.5 mg, 0.449 mmol), 2,4-difluorophenol (0.052
mL,
0.539 mmol), and cesium carbonate (293 mg, 0.898 mmol ) were placed in
microwave
vial, dissolved in acetonitrile (3 mL), and heated in a microwave reactor at
110 degrees
Celsius for 20 minutes. The mixture was cooled to room temperature and
concentrated
under vacuum, diluted with 1 N sodium hydroxide solution, and extracted three
times
with dichloromethane. The combined organic extracts were washed with brine,
dried
over sodium sulfate, filtered, and the filtrate was concentrated under vacuum.
The crude
material was purified by preparative reverse-phase HPLC on a Waters Atlantis
C18
column 4.6 x 50 mm, 0.005 mm eluting with a gradient of water in acetonitrile
(0.05 %
trifluoroacetic acid modifier) to give isopropyl 4-{5-cyano-4-[(2,4-
difluorophenoxy)methy1]-1H-pyrazol-1-yllpiperidine-1-carboxylate. Analytical
LCMS:
retention time: 3.62 minutes (Waters Atlantis C18 4.6 x 50 mm, 0.005 mm; 95%
water/acetonitrile linear gradient to 5% water/acetonitrile over 4.0min; 0.05
%
trifluoroacetic acid modifier; flow rate 2.0mUminute); LCMS (ES +): 405.18 (M
+ H).
Example 12: Isopropyl 4-{5-cyano-4-1-(2-methylphenoxy)methy11-1H-pyrazol-1-
yllpiperidine-1-carboxylate
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ON
y/
N ___________________________________________ 0
To a stirred solution of ortho-cresol (21 mg, 0.19 mmol) and isopropyl 4-(5-
cyano-4-
((methylsulfonyloxy)methyl)-1H-pyrazol-1-y1)piperidine-1-carboxylate
(Preparation 10)
(60 mg, 0.16 mmol) in acetonitrile (1.6 mL) was added cesium carbonate (106
mg, 0.32
mmol). The mixture was heated at reflux for 15 hours. After cooling to room
temperature the crude material was concentrated to dryness in vacuo, and the
residue
was taken up in water and extracted 3 times with ethyl acetate (20 mL each
extraction). The combined organic extracts were washed with brine, dried over
sodium
sulfate, filtered and the filtrate was concentrated to dryness under vacuum to
give a tan
residue (0.065 g, 100%). The crude sample was dissolved in dimethyl sulfoxide
(1 mL)
and purified by preparative reverse phase HPLC on a Waters Sunfire C1819 x 100
mm,
0.005 mm column, eluting with a linear gradient of 80% water/acetonitrile to
0%
water/acetonitrile in 8.5 minutes, followed by a 1.5 minute period at 0%
water/acetonitrile (0.05% trifluoroacetic acid modifier); flow
rate: 25mL/minute. Analytical LCMS: retention time 3.82 minutes (Waters
Atlantis C18
4.6 x 50 mm, 0.005 mm column; 95% water/acetonitrile linear gradient to 5%
water/acetonitrile over 4.0 minutes, followed by a 1 minute period at 5%
water/acetonitrile; 0.05% trifluoroacetic acid modifier; flow rate: 2.0
mL/minute); LCMS
(ES-I-) 383.2 (M+1).
Example 13: 1-Methylcyclopropyl 4-{5-cyano-4-[(2,5-difluorophenoxy)methy11-1H-
pyrazol-1-yllpiperidine-1-carboxylate
// ________________________________________
F * 1( p
N N-\
_______________________________________________ 0-7.<
A) tert-butyl 4-(5-cyano-4-((2,5-difluorophenoxy)methyl)-1H-pyrazol-1-
y1)piperidine-1-
carboxylate
To a stirred solution of 2,5-difluorophenol (54 mg, 0.39 mmol) and tert-butyl
4-(5-cyano-
4-((methylsulfonyloxy)methyl)-1H-pyrazol-1-y1)piperidine-1-carboxylate
(Preparation 16)
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(126 mg, 0.33 mmol) in 3 mL of acetonitrile was added cesium carbonate (214
mg, 0.66
mmol). The mixture was heated at reflux for 15 hours. The mixture was cooled
to room
temperature and diluted with ethyl acetate and water. The layers were
separated and
the aqueous phase was extracted with ethyl acetate. The combined organic
phases
were washed with brine, dried over magnesium sulfate, filtered, and the
filtrate was
concentrated in vacuo to give tert-butyl 4-(5-cyano-4-((2,5-
difluorophenoxy)methyl)-1H-
pyrazol-1-y1)piperidine-1-carboxylate which was used in the next step without
purification.
B) 4-((2,5-Difluorophenoxy)methyl)-1-(piperidin-4-y1)-1H-pyrazole-5-
carbonitrile
To a solution of tert-butyl 4-(5-cyano-4-((2,5-difluorophenoxy)methyl)-1H-
pyrazol-1-
y1)piperidine-1-carboxylate (137 mg, 0.33 mmol) in 5 mL of dichloromethane was
added
0.82 mL of hydrochloric acid (4 M in 1,4-dioxane). The mixture was stirred at
room
temperature for 2 hours before the mixture was concentrated in vacuo to give 4-
((2,5-
difluorophenoxy)methyl)-1-(piperidin-4-y1)-1H-pyrazole-5-carbonitrile which
was used in
the next step without purification.
C) 1-Methylcyclopropyl 4-{5-cyano-4-1-(2,5-difluorophenoxy)methy11-1H-pyrazol-
1-
yllpiperidine-1-carboxylate
To a stirred solution of 4-((2,5-difluorophenoxy)methyl)-1-(piperidin-4-y1)-1H-
pyrazole-5-
carbonitrile (104 mg, 0.33 mmol) in 3.3 mL of dichloromethane was added
triethylamine
(0.18 mL, 1.3 mmol) followed by 1-methylcyclopropyl 4-nitrophenyl carbonate
(see
Preparation 26 and W009105717) (171 mg, 0.72 mmol) at room temperature. The
resulting bright yellow mixture was stirred for 15 hours under a nitrogen
atmosphere. The reaction mixture was diluted with dichloromethane and water.
The
layers were separated and the aqueous phase was extracted with
dichloromethane.
The combined organic phases were washed with saturated aqueous sodium
bicarbonate, brine, dried over magnesium sulfate, filtered and the filtrate
was
concentrated in vacuo to give 225 mg of crude material. Part (45 mg) of this
material
was dissolved in dimethyl sulfoxide (0.9 mL) and purified by preparative
reverse-phase
HPLC on a Waters XBridge C18 column 19 x 100 mm, 0.005 column eluting with a
gradient of water in acetonitrile (0.03% ammonium hydroxide modifier).
Analytical
LCMS: retention time 3.60 minutes (Atlantis C18 4.6 x 50 mm, 5 micrometer
column;
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95% water/acetonitrile linear gradient to 5% water/acetonitrile over 4
minutes; 0.05%
trifluoroacetic modifier; flow rate 2.0 mUminute; LCMS (ES+): 417.1 (M+H).
Example 14: 1-Methylcyclopropyl 4-{5-cyano-4-1-(2,3-difluorophenoxy)methy11-1H-
oyrazol-1-ylloicieridine-1-carboxylate
F
0- \ 0
'N / 0
..... ,NI _____________________________ ( N¨ (\
The title compound was prepared using commercially available 2,3-
diflurophenol,
following procedures analogous to Example 13. The crude material (49 mg) was
dissolved in dimethyl sulfoxide (0.9 mL) and purified by preparative reverse-
phase
HPLC on a Waters XBridge C18 column 19 x 100 mm, 0.005 column eluting with a
gradient of water in acetonitrile (0.03% ammonium hydroxide modifier).
Analytical
LCMS: retention time 3.62 minutes (Atlantis C18 4.6 x 50 mm, 5 micrometer
column;
95% water/acetonitrile linear gradient to 5% water/acetonitrile over 4
minutes; 0.05%
trifluoroacetic modifier; flow rate 2.0 mUminute; LCMS (ES+): 417.2 (M+H).
Example 15: 1-Methylcyclopropyl 4-{4-[(4-carbamoy1-2-fluorophenoxy)methy1]-5-
cyano-
1H-oyrazol-1-ylloicieridine-1-carboxylate
NH2
0
* F
N
0...el
---N / 0
A) tert-Butyl 4-(4-((4-carbamoy1-2-fluoroohenoxy)methyl)-5-cyano-1H-oyrazol-1-
yl)piperidine-1-carboxylate
To a stirred solution of tert-butyl 4-(5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yl)piperidine-1-carboxylate (Preparation 15) (200 mg, 0.65 mmol), 3-fluoro-4-
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hydroxybenzamide (Preparation 23) (100 mg, 0.64 mmol) and triphenylphosphine
(188
mg, 0.72 mmol) in 3 mL of 1,4-dioxane was added drop-wise diethyl
azodicarboxylate
(0.11 mL, 0.69 mmol). The resulting mixture was stirred overnight at room
temperature
before the mixture was concentrated in vacuo. The residue was purified by
flash
chromatography, eluting with a gradient of 30 to 70% ethyl acetate in heptane
to give
tert-butyl 4-(4-((4-carbamoy1-2-fluorophenoxy)methyl)-5-cyano-1H-pyrazol-1-
yl)piperidine-1-carboxylate as a white solid (215 mg).
B) 4-((5-Cyano-1-(piperidin-4-y1)-1H-pyrazol-4-yl)methoxy)-3-fluorobenzamide
To a stirred solution of tert-butyl 4-(4-((4-carbamoy1-2-fluorophenoxy)methyl)-
5-cyano-
1H-pyrazol-1-yl)piperidine-1-carboxylate (215 mg, 0.48 mmol) in 2 mL of
dichloromethane was added 1 mL of trifluoroacetic acid at room temperature.
After 1
hour the solution was concentrated in vacuo. The residue was purified by flash
chromatography, eluting with a gradient mixture of 1 to 15% of methanol in
dichloromethane containing 2% of aqueous ammonia) to give 4-((5-cyano-1-
(piperidin-
4-y1)-1H-pyrazol-4-yl)methoxy)-3-fluorobenzamide as a white solid (150 mg).
C) 1-Methylcyclopropyl 4-{4-1-(4-carbamoy1-2-fluorophenoxy)methy11-5-cyano-1H-
pyrazol-1-yllpiperidine-1-carboxylate
To a stirred solution of 4-((5-cyano-1-(piperidin-4-y1)-1H-pyrazol-4-
yl)methoxy)-3-
fluorobenzamide (40 mg, 0.12 mmol) in 1 mL of dichloromethane was added
triethylamine (0.036 mL, 0.26 mmol), followed by 1-methylcyclopropyl 4-
nitrophenyl
carbonate (Preparation 26 and W009105717) (60 mg, 0.26 mmol) at room
temperature. The resulting bright yellow mixture was stirred for 2 hours under
a nitrogen
atmosphere at 65 degrees Celsius. The reaction was cooled to room temperature,
diluted with water and extracted twice with dichloromethane. The combined
organic
extracts were washed with saturated sodium bicarbonate, dried over sodium
sulfate,
filtered and the filtrate was concentrated in vacuo. The residue was purified
by flash
chromatography, eluting with a gradient of 40 to 90% ethyl acetate in heptane
to give 1-
methylcyclopropyl 4-{4-[(4-carbamoy1-2-fluorophenoxy)methyl]-5-cyano-1H-
pyrazol-1-
yllpiperidine-1-carboxylate as white solid (34 mg). 1H NMR (400 MHz,
deuterochloroform) delta 0.59 - 0.67 (m, 2 H), 0.83 - 0.92 (m, 2 H), 1.54 (s,
3 H), 2.02
(d, J=4.10 Hz, 2 H), 2.04 - 2.22 (m, 2 H), 2.91 (br. s., 2 H), 4.11 -4.43 (m,
2 H), 4.44 -
4.55 (m, 1 H), 5.15 (s, 2 H), 7.03 - 7.10 (m, 1 H), 7.52 - 7.62 (m, 2 H), 7.68
(s, 1 H). 1H
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NMR indicated the presence of less than 10% of what is believed to be the
corresponding isopropyl carbamate derivative (from the isopropyl 4-nitrophenyl
carbonate contaminating the 1-methylcyclopropyl 4-nitrophenyl carbonate). LCMS
(ES)
442.4 (M+1).
Example 16: 1-Methylcyclopropyl 4-{4-1-(4-carbamoylphenoxy)methy11-5-cyano-1H-
pyrazol-1-yllpiperidine-1-carboxylate
NH2
0, N
\ 0-e
The title compound was prepared using commercially available 4-
hydroxybenzamide,
following procedures analogous to Example 15. 1H NMR (400 MHz,
deuterochloroform)
delta 0.57 - 0.67 (m, 2 H), 0.84 - 0.91 (m, 2 H), 1.56 (s, 3 H), 1.93 - 2.05
(m, 2 H), 2.05 -
2.19 (m, 2 H), 2.91 (t, J=15.62 Hz, 2 H), 4.26 (br. s., 2 H), 4.44 - 4.55 (m,
1 H), 5.09 (s,
2 H), 6.96 - 7.04 (m, 2 H), 7.66 (s, 1 H), 7.75 - 7.82 (m, 2 H). 1H NMR
indicated the
presence of less than 10% of what is believed to be the corresponding
isopropyl
carbamate derivative (from the isopropyl 4-nitrophenyl carbonate contaminating
the 1-
methylcyclopropyl 4-nitrophenyl carbonate). LCMS (ES) 424.4 (M+1).
Example 17: 1-Methylcyclopropyl 4-(5-cyano-4-((4-cyanophenoxy)methyl)-1H-
pyrazol-1-
yl)piperidine-1-carboxylate
NC
O N
0-e
,N-CN-µ
--"N / 0
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The title compound was prepared using commercially available 4-
hydroxybenzonitrile,
following procedures analogous to Example 15. The purification of the crude
reaction
mixture was performed by flash chromatography, eluting with a gradient mixture
of ethyl
acetate in heptane (0 to 100% ethyl acetate). 1H NMR (500 MHz,
deuterochloroform)
delta 0.60 - 0.70 (m, 2 H), 0.84 - 0.94 (m, 2 H), 1.23- 1.31 (m, 1 H), 1.56
(s, 3 H), 2.01 -
2.15(m, 4 H), 2.93(m, 2 H), 4.11 - 4.37 (m, 1 H), 4.49 - 4.55 (m, 1 H),
5.10(s, 2 H),
7.03 (d, J=8.78 Hz, 2 H), 7.63 (d, J=8.78 Hz, 2 H), 7.67 (s, 1 H).
Example 18: Isopropyl 4-(4-((4-(1H-pyrazol-1-yl)phenoxy)methyl)-5-cyano-1H-
pyrazol-1-
yl)piperidine-1-carboxylate
aii
ON
U/
'z-----NI 0
The title compound was prepared using 4-(1H-pyrazol-1-yl)phenol (WO
2003072547),
following a procedure analogous to Example 12. The purification of the crude
reaction
mixture was performed by flash chromatography, eluting with a gradient mixture
of ethyl
acetate in heptane (0 to 100% ethyl acetate). 1H NMR (500 MHz,
deuterochloroform)
delta 1.28 (d, J=6.34 Hz, 6 H), 2.01 -2.09 (m, 2 H), 2.17 (m, 2 H), 2.91 -2.99
(m, 2 H),
4.37 (m, 2 H), 4.50 - 4.58 (m, 1 H), 4.93-4.98 (m, 1 H), 5.11 (s, 2 H), 6.47
(t, J=2.07 Hz,
1 H), 7.07 (d, J=9.03 Hz, 2 H), 7.64 (d, J=9.03 Hz, 2 H), 7.70 (s, 1 H), 7.72
(d, J=1.71
Hz, 1 H), 7.86 (d, J=2.44 Hz, 1 H). LCMS (ES) 435.4(M+1).
Example 19: Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1H-tetrazol-5-
yl)phenoxy)methyl)-1H-
pyrazol-1-yl)piperidine-1-carboxylate and Isopropyl 4-(5-cyano-4-((2-fluoro-4-
(2H-
tetrazol-5-yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
93
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. N.
N',N NH
HN' 'N
'NI¨ 1\1¨
40 N 441k N
0....õ.et
U/
F N \ F 0¨(
N--(
/
....
----N 0 --N / 0
A) Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-((2-(trimethylsilypethoxy)methyl)-1H-
tetrazol-5-
y1)phenoxy)methyl)-1H-pyrazol-1-y1)piperidine-1-carboxylate and Isopropyl 4-(5-
cyano-
4-((2-fluoro-4-(2-((2-(trimethylsilypethoxy)methyl)-2H-tetrazol-5-
y1)phenoxy)methyl)-1H-
pyrazol-1-yl)piperidine-1-carboxylate
To a stirred solution of isopropyl 4-(5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yl)piperidine-1-carboxylate (94 mg, 0.322 mmol), 2-fluoro-4-(1-((2-
(trimethylsilypethoxy)methyl)-1H-tetrazol-5-y1)phenol and 2-fluoro-4-(2-((2-
(trimethylsilypethoxy)methyl)-2H-tetrazol-5-y1)phenol (Preparation 17) (100
mg, 0.322
mmol) and triphenylphosphine (110 mg, 0.42 mmol) in 5 mL of 1,4-dioxane was
added
drop-wise diethyl azodicarboxylate (0.060 mL, 0.39 mmol). The resulting
mixture was
stirred overnight at room temperature before the mixture was concentrated in
vacuo.
The residue was purified by flash chromatography, eluting with a gradient of
10 to 40%
ethyl acetate in heptane to give isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-((2-
(trimethylsilyl)ethoxy)methyl)-1H-tetrazol-5-yl)phenoxy)methyl)-1H-pyrazol-1-
yl)piperidine-1-carboxylate and isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-((2-
(trimethylsilypethoxy)methyl)-2H-tetrazol-5-y1)phenoxy)methyl)-1H-pyrazol-1-
y1)piperidine-1-carboxylate (140 mg, 74% yield).
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-((2-(trimethylsilypethoxy)methyl)-2H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate.1H NMR (400 MHz,
deuterochloroform) delta -0.05-0.01 (m, 9 H), 0.90- 1.00 (m, 2 H), 1.18 - 1.27
(m, 6 H),
2.02 (br. s., 2 H), 2.13 (m, 2 H) 2.93 (br. s., 2 H), 3.65- 3.78 (m, 2 H),
4.30 (d, J=7.22
Hz, 2 H), 4.46 - 4.58 (m, 1 H), 4.86 - 4.98 (m, 1 H), 5.16 (s, 2 H), 5.89 (s,
2 H), 7.09 -
7.18 (m, 1 H), 7.69 (s, 1 H), 7.88 - 7.96 (m, 2 H). LCMS (ES) 585.1 (M+1).
B) Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1H-tetrazol-5-yl)phenoxy)methyl)-1H-
pyrazol-1-
yl)piperidine-1-carboxylate and Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
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Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-((2-(trimethylsilypethoxy)methyl)-1H-
tetrazol-5-
y1)phenoxy)methyl)-1H-pyrazol-1-y1)piperidine-1-carboxylate and isopropyl 4-(5-
cyano-
4-((2-fluoro-4-(2-((2-(trimethylsilypethoxy)methyl)-2H-tetrazol-5-
y1)phenoxy)methyl)-1H-
pyrazol-1-y1)piperidine-1-carboxylate (220 mg, 0.38 mmol) were dissolved in
ethanol (3
mL) and a solution of aqueous 2 M hydrochloric acid (3 mL) was added drop-
wise, The
resulting mixture was stirred at 50 degrees Celsius for 4 hours before being
cooled
down to room temperature and filtered. The resulting white solid was washed
with ethyl
acetate and heptane (1/1 volume) and dried under reduced pressure to give the
title
compound (80 mg, 47% yield). 1H NMR (400 MHz, deutero dimethyl sulfoxide)
delta
1.16 (d, J=6.25 Hz, 6 H), 1.76 - 1.90 (m, 2 H), 1.98 (dd, J=14.45, 3.12 Hz, 2
H), 2.99 (br.
s., 2 H), 4.04 (d, J=15.81 Hz, 2 H), 4.59 - 4.71 (m, 1 H), 4.70 - 4.82 (m, 1
H), 5.27 (s, 2
H), 7.47 - 7.57 (m, 1 H), 7.80 - 7.83 (m, 1 H), 7.83 - 7.87 (m, 1 H), 7.90 (s,
1 H). LCMS
(ES) 455.0 (M+1).
Example 20: Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-methyl-1H-tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate and
Example 21: Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-methyl-2H-tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
N,
N'' N--- N,
-----N' ' N
1\1-
41 N i\l-
410 N
0....1
F F I
N-( ________________________________________________________________
\ 0-(
/
'N 0 ---=N'
/ 0
To a solution of isopropyl 4-(5-cyano-4-((2-fluoro-4-(1H-tetrazol-5-
yl)phenoxy)methyl)-
1H-pyrazol-1-yl)piperidine-1-carboxylate and isopropyl 4-(5-cyano-4-((2-fluoro-
4-(2H-
tetrazol-5-yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (70 mg,
0.15
mmol) at room temperature in tetrahydrofuran (2 mL) was added sodium hydride
(14
mg, 0.31 mmol) in two portions, and the resulting mixture was stirred for 5
minutes.
lodomethane (0.03 mL, 0.46 mmol) was then added and the reaction mixture was
stirred at room temperature for an additional 16 hours. The reaction was
quenched by
addition of water and the mixture was diluted with ethyl acetate. The organic
phase was
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separated and the aqueous phase was extracted twice with ethyl acetate. The
combined organic extracts were washed with brine, dried over magnesium
sulfate,
filtered and the filtrate was concentrated in vacuo. The residue was purified
by flash
silica gel chromatography, eluting with a gradient mixture of ethyl acetate in
heptane (30
to 60% ethyl acetate) to give isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-methyl-1H-
tetrazol-
5-yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (10 mg, 14%
yield) and
isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-methyl-2H-tetrazol-5-yl)phenoxy)methyl)-
1H-
pyrazol-1-yl)piperidine-1-carboxylate (30 mg, 42% yield).
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-methyl-1H-tetrazol-5-yl)phenoxy)methyl)-
1H-
pyrazol-1-yl)piperidine-1-carboxylate (Example 20). 1H NMR (400 MHz,
deuterochloroform) delta 1.18- 1.28 (m, 6 H), 1.95 - 2.06 (m, 2 H), 2.13 (m, 2
H), 2.85 -
3.02 (m, 2 H), 4.17 (s, 3 H), 4.36 (d, J=10.15 Hz, 2 H), 4.46 - 4.57 (m, 1 H)
4.92 (spt, 1
H), 5.19 (s, 2 H), 7.17- 7.24 (m, 1 H), 7.48 - 7.58 (m, 2 H), 7.70 (s, 1 H).
LCMS (ES)
469.0 (M+1).
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-methyl-2H-tetrazol-5-yl)phenoxy)methyl)-
1H-
pyrazol-1-yl)piperidine-1-carboxylate (Example 21). 1H NMR (400 MHz,
deuterochloroform) delta 1.24 (d, J=6.25 Hz, 6 H) 1.95 - 2.05 (m, 2 H) 2.13
(m, 2 H)
2.93 (t, J=12.59 Hz, 2 H) 4.31 (br. s., 2 H) 4.37 (s, 3 H) 4.51 (m, 1 H) 4.92
(m, 1 H) 5.16
(s, 2 H) 7.09 - 7.16 (m, 1 H) 7.69 (s, 1 H) 7.83 - 7.87 (m, 1 H) 7.87- 7.90
(m, 1 H).
LCMS (ES) 469.0 (M+1).
Example 22: Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-(2-hydroxyethyl)-2H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
N,õ,
HO---/---N' 11
i\l-
fa N
F C(N...e...//
/ ______________________________________________ \ 0
'NI ____________________________________________ / 0
A) Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-(2-(trimethylsilyloxy)ethyl)-2H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
To a stirred solution of isopropyl 4-(5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yl)piperidine-1-carboxylate (Preparation 5) (78 mg, 0.266 mmol), 2-fluoro-4-(2-
(2-
(trimethylsilyloxy)ethyl)-2H-tetrazol-5-yl)phenol (Preparation 19) (90 mg,
0.27 mmol)
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and triphenylphosphine (77 mg, 0.29 mmol) in 5 mL of 1,4-dioxane was added
drop-
wise diethyl azodicarboxylate (0.046 mL, 0.28 mmol). The resulting mixture was
stirred
for 15 hours at room temperature before the mixture was concentrated in vacuo.
The
residue was purified by flash chromatography, eluting with a gradient of 5 to
40% ethyl
acetate in heptane to give isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-(2-
(trimethylsilyloxy)ethyl)-2H-tetrazol-5-yl)phenoxy)methyl)-1H-pyrazol-1-
yl)piperidine-1-
carboxylate (140 mg, 86% yield).
B) Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-(2-hydroxyethyl)-2H-tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(2-(2-(trimethylsilyloxy)ethyl)-2H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (140 mg, 0.228
mmol) was
dissolved in methanol (2 mL), and a solution of 4 M hydrochloric acid (1 mL)
in 1,4-
dioxane was added drop-wise, The resulting mixture was stirred at room
temperature
for 2 hours before being concentrated under reduced pressure. The residue (160
mg)
was divided and ca. 50 mg of the crude was purified by reverse-phase HPLC to
give the
title compound (30 mg, 26%) (Column: Waters XBridge C18 19x100, 5 micrometer;
Mobile phase A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03%
ammonium hydroxide in acetonitrile (v/v); Gradient: 85%water/15%acetonitrile
linear to
0%water/100%acetonitrile in 8.5 minutes, hold at 0%water / 100%acetonitrile to
10.0
minutes. Flow: 25mL/min. Detection: 215 nm. LCMS (ES+): 499.5 (M+1).
Example 23: Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-(2-hydroxyethyl)-1H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
r-OH
1\l'"N.,NJ
1\1-
10 N
F U/
--N / o
A) Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-(2-(trimethylsilyloxy)ethyl)-1H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
To a stirred solution of isopropyl 4-(5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yl)piperidine-1-carboxylate (43 mg, 0.15 mmol), 2-fluoro-4-(1-(2-
(trimethylsilyloxy)ethyl)-
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1H-tetrazol-5-yl)phenol (preparation 20) (50 mg, 0.15 mmol) and
triphenylphosphine (43
mg, 0.16 mmol) in 3 mL of 1,4-dioxane was added drop-wise diethyl
azodicarboxylate
(0.025 mL, 0.16 mmol). The resulting mixture was stirred overnight at room
temperature
before the mixture was concentrated in vacuo. The residue was purified by
flash
chromatography, eluting with a gradient of 30 to 70% ethyl acetate in heptane
to give
isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-(2-(trimethylsilyloxy)ethyl)-1H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (50 mg, 55%
yield).
B) Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-(2-hydroxyethyl)-1H-tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
Isopropyl 4-(5-cyano-4-((2-fluoro-4-(1-(2-(trimethylsilyloxy)ethyl)-1H-
tetrazol-5-
yl)phenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (50 mg, 0.082
mmol) was
dissolved in methanol (2 mL) and a solution of 4 M hydrochloric acid (1 mL) in
1,4-
dioxane was added drop-wise, The resulting mixture was stirred at room
temperature
for 2 hours before being concentrated under reduced pressure. The residue (60
mg)
was purified by reversed-phase HPLC to give the title compound (20 mg, 49%
yield)
(Column: Waters XBridge C18 19x100, 5 micrometer; Mobile phase A: 0.03%
ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium hydroxide in
acetonitrile (v/v); Gradient: 80%water/20%acetonitrile linear to
0%water/100%acetonitrile in 8.5 minutes,
hold at 0%water / 100%acetonitrile to 10.0 minutes. Flow: 25mL/min. Detection:
215
nm
LCMS (ES+): 499.4 (M+1).
Example 24: 1-Methylcyclopropyl 4-(5-cyano-4-{[2-fluoro-4-(1-methyl-1H-
tetrazol-5-
yl)phenoxy]methyll-1H-pyrazol-1-y1)piperidine-1-carboxylate
N
FN,
0 ___________________________________________________ (\
_(
N
/ 0
The title compound was prepared using 2-fluoro-4-(1-methyl-1H-tetrazol-5-
yl)phenol
(Preparation 21), following procedures analogous to Example 15. 1H NMR (400
MHz,
deuterochloroform) delta 0.58 - 0.67 (m, 2 H), 0.83 - 0.92 (m, 2 H), 1.57 (s,
3 H), 1.94 -
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2.05 (m, 2 H), 2.05 - 2.21 (m, 2 H), 2.92 (t, J=12.98 Hz, 2 H), 4.17 (s, 3 H),
4.32 (br. s.,
2 H), 4.43 - 4.56 (m, 1 H), 5.19 (s, 2 H), 7.17 - 7.24 (m, 1 H), 7.48 - 7.58
(m, 2 H), 7.70
(s, 1 H). 1H NMR indicated the presence of less than 10% of what is believed
to be the
corresponding isopropyl carbamate derivative (from the isopropyl 4-nitrophenyl
carbonate contaminating the 1-methylcyclopropyl 4-nitrophenyl carbonate). LCMS
(ES)
481.6 (M+1).
Example 25: 1-Methylcyclopropyl 4-(5-cyano-4-{1-4-(1-methy1-1H-tetrazol-5-
yl)phenoxylmethyll-1H-pyrazol-1-yl)piperidine-1-carboxylate
NN'
N,, "
N git iNi
0 ,- _< ______________________________________ \N_LC)-e
N
The title compound was prepared using 4-(1-methyl-1H-tetrazol-5-y1)phenol
(Preparation 22), following procedures analogous to Example 15.
1H NMR (400 MHz, deuterochloroform) delta 0.60 - 0.67 (m, 2 H), 0.83 - 0.91
(m, 2 H),
1.58 (s, 3 H), 1.96 - 2.06 (m, 2 H), 2.06 - 2.21 (m, 2 H), 2.84 - 3.00 (m, 2
H), 4.16 (s, 3
H), 4.33 (br. s., 2 H), 4.45 - 4.57 (m, 1 H), 5.12 (s, 2 H), 7.10 -7.15 (m, 2
H), 7.68 (s, 1
H), 7.69 - 7.74 (m, 2 H). 1H NMR indicated the presence of less than 10% of
what is
believed to be the corresponding isopropyl carbamate derivative (from the
isopropyl 4-
nitrophenyl carbonate contaminating the 1-methylcyclopropyl 4-nitrophenyl
carbonate).
LCMS (ES) 463.5 (M+1).
Example 26: 1-Methylcyclopropyl 4-(4-((4-carbamoy1-3-fluorophenoxy)methyl)-5-
cyano-
1H-pyrazol-1-yl)piperid ine-1-carboxylate
NH2 F
ID
fk N
0.õ..1
-- \ 0
,N1 / N-
(\
...-N \ 0
99
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The title compound was prepared using 2-fluoro-4-hydroxybenzamide (Preparation
24),
following procedures analogous to Example 13. 1H NMR (400 MHz,
deuterochloroform)
delta 0.57 - 0.65 (m, 2 H), 0.82 - 0.89 (m, 2 H), 1.53 (s, 3 H), 1.92 - 2.04
(m, 2 H), 2.10
(qd, J=12.14, 4.20 Hz, 2 H), 2.90 (br. s., 2 H), 4.32 (br. s., 2 H), 4.49 (tt,
J=11.25, 4.37
Hz, 1 H), 5.02 - 5.09 (m, 2 H), 6.00 (br. s., 1 H), 6.51 - 6.64 (m, 1 H), 6.69
(dd, J=13.66,
2.54 Hz, 1 H), 6.84 (dd, J=8.78, 2.54 Hz, 1 H), 7.64 (s, 1 H), 8.07 (t, J=9.08
Hz, 1 H). 1H
NMR indicated the presence of less than 10% of what is believed to be the
corresponding isopropyl carbamate derivative (from the isopropyl 4-nitrophenyl
carbonate contaminating the 1-methylcyclopropyl 4-nitrophenyl carbonate). LCMS
(ES)
442.4(M+1).
Example 27: Isopropyl 4-(5-cyano-4-{1-1-2-fluoro-4-
(methylsulfonyl)phenoxylethy11-1H-
pyrazol-1-y1)piperidine-1-carboxylate
0
ii
0
0
F
Y
NNo
N
0
The title compound was prepared using 2-fluoro-4-(methylsulfonyl)phenol and
isopropyl
4-(5-cyano-4-(1-hydroxyethyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate
(Preparation
25), following procedures analogous to Example 15. The sample was purified by
reversed-phase HPLC (Column: Waters XBridge C18 19x100, 5 micrometer; Mobile
phase A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03%
ammonium hydroxide in acetonitrile (v/v); Gradient: 80%water/20%acetonitrile
linear to
0%water/100%acetonitrile in 8.5 minutes, hold at 0%water / 100%acetonitrile to
10.0
minutes. Flow: 25mL/minute. LCMS ( ES+): 479.2 M+1).
Example 28: Isopropyl 4-(5-cyano-4-{1-1-(2-methylpyridin-3-yl)oxylethyll-1H-
pyrazol-1-
yl)piperidine-1-carboxylate
ON \i-i
/ 0
\ \
N N
100
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The title compound was prepared using 2-methylpyridin-3-ol and isopropyl 4-(5-
cyano-
4-(1-hydroxyethyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (Preparation 25) ,
following
procedures analogous to Example 15. The sample was purified by reversed-phase
HPLC (Column: Waters XBridge C18 19x100, 5 micrometer; Mobile phase A: 0.03%
ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium hydroxide in
acetonitrile (v/v); Gradient: 85%water/15%acetonitrile linear to
0%water/100%acetonitrile in 8.5 minutes, hold at 0%water / 100%acetonitrile to
10.0
minutes. Flow: 25mL/minute. LCMS (ES+): 398.2 M+1).
Example 29: Isopropyl 4-(5-cyano-4-{212-fluoro-4-
(methylsulfonyl)phenyl]propy11-1H-
pyrazol-1-yl)piperidine-1-carboxylate
,N _________________________________________
N
/ b
A) Isopropyl 4-(5-cyano-4-viny1-1H-pyrazol-1-yl)piperidine-1-carboxylate
To a stirred mixture of (methyl)-triphenylphosphonium bromide (323 mg, 0.88
mmol) in
tetrahydrofuran (5 mL) at -78 degrees Celsius was added drop-wise n-
butyllithium
(0.360 mL, 0.89 mmol, 2.5 M in hexanes). The resulting yellow mixture was
stirred at -
78 degrees Celsius for 30 minutes, and then a solution of isopropyl 4-(5-cyano-
4-formyl-
1H-pyrazol-1-yl)piperidine-1-carboxylate (Example 9, Step A) (171 mg, 0.59
mmol) in
tetrahydrofuran (2.5 mL) was added. The cold bath was removed, and the
reaction
mixture was stirred for 3.75 hours at room temperature. The reaction was
quenched
with saturated aqueous ammonium chloride, and the mixture was extracted twice
with
ethyl acetate. The combined extracts were washed sequentially with water and
brine
and then dried over sodium sulfate. The mixture was filtered, and the filtrate
was
concentrated in vacuo. The residue was purified by silica gel chromatography
eluting
with a gradient mixture of ethyl acetate in heptane (10 to 100%) to give the
title
compound as a clear oil (116 mg, 68%). 1H NMR (500 MHz, deuterochloroform)
delta
0.88 (d, J=6.10 Hz, 6 H), 1.55 - 1.67 (m, 2 H), 1.68 - 1.84 (m, 2 H), 2.43 -
2.73 (m, 2 H),
3.95 (br. s., 2 H), 4.04 - 4.21 (m, 1 H) 4.44 - 4.67 (m, 1 H), 5.02 (d,
J=11.22 Hz, 1 H),
5.43 (d, J=17.81 Hz, 1 H), 6.20 (dd, J=17.81, 11.22 Hz, 1 H), 7.27 (s, 1 H).
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B) (E,Z)-Isopropyl 4-(5-cyano-4-(2-(2-fluoro-4-(methylsulfonyl)phenyl)prop-1-
eny1)-1H-
pyrazol-1-yl)piperidine-1-carboxylate
To a solution of isopropyl 4-(5-cyano-4-vinyl-1H-pyrazol-1-yl)piperidine-1-
carboxylate
(116 mg, 0.4 mmol) and 2-fluoro-4-(methylsulfonyI)-1-(prop-1-en-2-yl)benzene
(Preparation 29) (43 mg, 0.20 mmol) in anhydrous dichloromethane (2 mL) was
added
the second generation Hoveyda-Grubbs catalyst (commercially available from
Aldrich)
(12.5 mg, 0.020 mmol). The green solution was heated at 40 degrees Celsius for
72
hours periodically adding dichloromethane. The material was concentrated under
reduced pressure, and the residue purified by silica gel chromatography (10 to
100%
ethyl acetate in heptane) to give the product as an impure oil (8 mg, 8%).
This material
was used as is. LCMS (APCI): 473.2 (M ¨ 1).
C) Isopropyl 4-(5-cyano-4-{2-1-2-fluoro-4-(methylsulfonyl)phenyllpropy11-1H-
pyrazol-1-
yl)piperidine-1-carboxylate
A solution of (E,Z)-isopropyl 4-(5-cyano-4-(2-(2-fluoro-4-(methylsulfonyI)-
phenyl)prop-1-
enyI)-1H-pyrazol-1-yl)piperidine-1-carboxylate (8 mg, 0.02 mmol) in ethyl
acetate (3 mL)
was hydrogenated on the H-Cube TM at the "full hydrogen" setting using a 10%
palladium
on carbon cartridge at a flow rate of 1 mL/minute. The material was
concentrated in
vacuo, and the residue (4 mg) was purified by reversed-phase HPLC (Column:
Waters
XBridge C18 19x100, 5 micrometer; Mobile phase A: 0.03% ammonium hydroxide in
water (v/v); Mobile phase B: 0.03% ammonium hydroxide in acetonitrile (v/v);
Gradient: 80%water/ 20%acetonitrile linear to 0%water/100%acetonitrile in 8.5
minutes,
hold at 0% water! 100% acetonitrile to 10.0 minutes. Flow: 25mL/minute) to
give the
title compound (1.9 mg, 23%): LCMS (ES+): 477.2 (M+1).
Example 30: 1-Methylcyclopropyl 4-(5-cyano-4-{[(2-methylpyridin-3-
yl)oxy]methyll-1H-
pyrazol-1-y1)piperidine-1-carboxylate
I N
\
0_,/
/
---1\1' 0
The title compound was prepared using 2-methylpyridin-3-ol, following
procedures
analogous to Example 13. The crude material was purified by flash
chromatography,
eluting with a gradient mixture of ethyl acetate in heptane (60 to 100% ethyl
acetate) to
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give 77 mg of the title compound as a white solid. 1H NMR (400 MHz,
deuterochloroform) delta 0.60 - 0.66 (m, 2 H), 0.83 - 0.90 (m, 2 H), 1.55 (s,
3 H), 1.96 -
2.05 (m, 2 H), 2.05 - 2.20 (m, 2 H), 2.49 (s, 3 H), 2.84 - 2.98 (m, 2 H), 4.11
- 4.42 (m, 2
H), 4.46 - 4.55 (m, 1 H), 5.04 (s, 2 H), 7.06 - 7.16 (m, 2 H), 7.65 (s, 1 H),
8.12 (dd,
J=4.49, 1.56 Hz, 1 H).
Example 31: 1-Methylcyclopropyl 4-{5-cyano-4-[(2,3,6-trifluorophenoxy)methyI]-
1H-
pyrazol-1-yllpiperidine-1-carboxylate
F
lei F
N
0...õ1/
F
__________________________________ / -""-----N' 0
A) tert-Butyl 4-(5-cyano-4-((2,3,6-trifluorophenoxy)methyl)-1H-pyrazol-1-
y1)piperidine-1-
carboxylate
tert-Butyl 4-(5-cyano-4-((methylsulfonyloxy)methyl)-1H-pyrazol-1-y1)piperidine-
1-
carboxylate (Preparation 16) (87.8 mg, 0.228 mmol), 2,3,6-trifluorophenol
(51.7 mg,
0.342 mmol), and cesium carbonate (149 mg, 0.456 mmol) were placed in
microwave
vial and dissolved in acetonitrile (3 mL). The vial was heated in a microwave
reactor at
110 degrees Celsius for 20 minutes. The mixture was concentrated under reduced
pressure, and the residue was taken up in 1 N sodium hydroxide solution (5 mL)
and
extracted three times with dichloromethane. The combined organic extracts were
washed with brine, dried over sodium sulfate, filtered, and the filtrate was
concentrated
under reduced pressure. The crude material was purified by chromatography
eluting
with a 0 to 30 % ethyl acetate in heptane gradient to give 36.2 mg of tert-
butyl 4-(5-
cyano-4-((2,3,6-trifluorophenoxy)methyl)-1H-pyrazol-1-yl)piperidine-1-
carboxylate as a
clear oil.
1111-Methylcyclopropyl 4-{5-cyano-4-1-(2,3,6-trifluorophenoxy)methy11-1H-
pyrazol-1-
yllpiperidine-1-carboxylate
1-Methylcyclopropyl 4-{5-cyano-4-[(2,3,6-trifluorophenoxy)methyI]-1H-pyrazol-1-
yllpiperidine-1-carboxylate was prepared using commercially available 2,3,6-
trifluoro
phenol, following procedures analogous to Example 13 (B and C). The crude
material
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(17.1 mg) was purified by preparative reverse-phase HPLC on a Sepax 2-Ethyl
Pyridine column 250 x 21.2 mm, 0.005 eluting with a gradient of ethanol in
heptane.
Analytical LCMS: retention time 11.769 minutes (Phenomenex Luna (2) C18 150 x
3.0mm, 5 micrometer column; 95% water/methanol linear gradient to 100%
methanol
over 12.5 minutes; 0.1% formic acid modifier; flow rate 0.75 mUminute; LCMS
(ES+):
456.9 ( M + Na). 1H NMR (500 MHz, deuterochloroform) delta 0.64 - 0.66 (m, 2
H), 0.88
- 0.91 (m, 2 H), 1.57 (s, 3 H), 2.00 (d, J=10.49 Hz, 2 H), 2.07 - 2.18 (m, 2
H), 2.91 -
2.95 (m, 2 H), 4.18 (br. s., 1 H), 4.36 (br. s., 1 H), 4.50 (tt, J=11.34, 4.15
Hz, 1 H), 5.19
(s, 2 H), 6.83 - 6.90 (m, 2 H), 7.67 (s, 1 H).
Example 32: Isopropyl 4-{5-cyano-4-[(2,3,6-trifluorophenoxy)methyI]-1H-pyrazol-
1-
yllpiperidine-1-carboxylate
F
40 F
N
Oc_e_t
F
'N / 0
The title compound was prepared using commercially available 2,3,6-
trifulorophenol
following procedures analogous to Example 11. The crude material was purified
by
column chromatography eluting with a 0 to 25% ethyl acetate in heptane
gradient to
give isopropyl 4-{5-cyano-4-[(2,3,6-trifluorophenoxy)methyI]-1H-pyrazol-1-
yllpiperidine-
1-carboxylate as a clear oil. 1H NMR (500 MHz, deuterochloroform) delta 1.26
(d,
J=6.10 Hz, 6 H), 2.01 (d, J=11.22 Hz, 2 H) 2.13 (qd, J=12.28, 4.64 Hz, 2 H),
2.88 - 3.01
(m, 2 H), 4.32 (br. s., 2 H) 4.51 (tt, J=11.34, 4.15 Hz, 1 H), 4.90 - 4.98
(m,1 H), 5.18(s,
2 H), 6.82 - 6.92 (m, 2 H), 7.67 (s, 1 H); LCMS (ES+): 423.4 ( M + H).
Example 33: Isopropyl 4-(5-cyano-4-{12-fluoro-4-(1-methyl-1H-imidazol-2-
yl)phenoxylmethyll-1H-pyrazol-1-yl)piperidine-1-carboxylate
(N,
N' 41 F
N
N
'N' -( ______________________________________ / \ 0-(
N(
0
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The title compound was prepared from 2-fluoro-4-(1-methyl-1H-imidazol-2-
yl)phenol
(Preparation 28) and isopropyl 4-(5-cyano-4-((methylsulfonyloxy)methyl)-1H-
pyrazol-1-
y1)piperidine-1-carboxylate (Preparation 10) following procedures analogous to
Example
11. The crude material was purified by preparative reverse-phase HPLC on a
Sepax
Example 34: Isopropyl 4-(5-cyano-4-{1-2-fluoro-4-(1-methyl-1H-imidazol-5-
15 yl)phenoxylmethy11-1H-pyrazol-1-y1)piperidine-1-carboxylate
N
F
\ 0
0
The title compound was prepared from 2-fluoro-4-(1-methyl-1H-imidazol-5-
yl)phenol
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Example 35: Isopropyl 4-1-5-cyano-4-({12-methy1-6-(1H-1,2,4-triazol-1-
yl)pyridin-3-
ylloxylmethyl)-1H-pyrazol-1-yllnineridine-1-carboxylate
N,-.....1
µNi-NN,
, I N
/ ___________________________________________ \ 0-(
N- /N-
-14 _______ 0
The title compound was prepared using 2-methyl-6-(1H-1,2,4-triazol-1-
y1)pyridin-3-ol
following procedures analogous to Example 12. The sample was purified by
reversed-
phase HPLC (Column: Waters XBridge C18 19x100, 5 micrometer; Mobile phase
A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium
hyrdroxide in acetonitrile (v/v); Gradient: 80%water/20%acetonitrile linear to
0%water/100%acetonitrile in 8.0 minutes, hold at 0% water! 100% acetonitrile
to 9.5
minutes. Flow: 25mL/minute. LCMS (MS ES+:451.1).
Example 36: Isopropyl 4-1-5-cyano-4-({12-methy1-6-(1H-1,2,4-triazol-1-
yl)pyridin-3-
yllaminolmethyl)-1H-pyrazol-1-yllpiperidine-1-carboxylate
Nz-1
...N N
I N
NI--1/
\ 0-(
,N-( N __________________________________________ =K
/ \
'N ______________________________________________ o
To a stirred solution of isopropyl 4-(5-cyano-4-((methylsulfonyloxy)methyl)-1H-
pyrazol-
1-y1)piperidine-1-carboxylate (Preparation 10) (44 mg, 0.12 mmol) in 0.75 mL
of
tetrahydrofuran was added N,N-diisopropylethylamine (0.042 mL, 0.24 mmol)
followed
by 2-methyl-6-(1H-1,2,4-triazol-1-y1)pyridin-3-amine (21 mg, 0.12 mmol). The
reaction
mixture was heated at 60 degrees Celsius for 16 hours before it was cooled to
room
temperature and diluted with water and brine. The mixture was then extracted
three
times with 15 mL ethyl acetate. The combined organic extracts were washed with
brine,
dried over sodium sulfate, filtered and the filtrate was concentrated in vacuo
to give 52
mg of a yellow foam. The sample was purified by reversed-phase HPLC (Column:
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Waters Sunfire C18 19x100, 5 micrometer; Mobile phase A: 0.05% trifluoroacetic
acid
in water (v/v); Mobile phase B: 0.05% trifluoroacetic acid in acetonitrile
(v/v));
Gradient: 90%water/10%acetonitrile linear to 0%water/100%acetonitrile in 8.5
minutes,
hold at 0% water! 100% acetonitrile to 10.0 minutes. Flow: 25mL/minute. LCMS
(MS
ES+: 450.1).
Example 37: Isopropyl 415-cyano-4-({[2-methyl-6-(methylsulfonyl)pyridin-3-
yllaminolmethyl)-1H-pyrazol-1-yllpiperidine-1-carboxylate
0
ii
, \ix
0's 1
N
1
NI-t
\N 0-K
-µ
--11 / 0
The title compound was prepared using 2-methyl-6-(methylsulfonyl)pyridin-3-
amine
following procedures analogous to Example 36. The sample was purified by
reversed-
phase HPLC (Column: Waters XBridge C18 19x100, 5 micrometer; Mobile phase
A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium
hyrdroxide in acetonitrile (v/v); Gradient: 85%water/15%acetonitrile linear to
0%water/100%acetonitrile in 8.5 minutes, hold at 0% water! 100% acetonitrile
to 10.0
minutes. Flow: 25mL/minute. LCMS (ES+): 461.0 (M+1).
Example 38: 1-Methylcyclopropyl 4-(5-cyano-4-{1-4-(1H-tetrazol-1-
yl)phenoxylmethyll-
1H-pyrazol-1-yl)piperidine-1-carboxylate
,1*---:---N
/ k
N ,
N 0
N
I I 0-'1
0
-N
The title compound was prepared using commercially available 4-tetrazol-1-yl-
phenol
following procedures analogous to Example 15. The crude material was purified
by
flash chromatography eluting with a gradient from 0% to 75% ethyl acetate in
heptanes.
1H NMR (400 MHz, deuterochloroform) delta ppm 0.60 - 0.66 (m, 2 H) 0.84 - 0.90
(m, 2
H) 1.19 (t, J=7.03 Hz, 1 H) 1.55 (s, 3 H) 2.03 (br. s., 2 H) 2.06 - 2.19 (m, 2
H) 2.92 (br. s.,
2 H) 3.46 (q, J=7.09 Hz, 1 H) 4.46 -4.56 (m, 1 H) 5.11 (s, 2 H) 7.11 -7.16 (m,
2 H) 7.60
- 7.65 (m, 2 H) 7.68 (s, 1 H) 8.90 (s, 1 H)
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Example 39: 1-0-(5-Ethylpyrimidin-2-yl)piperidin-4-y11-4-{1-4-(1H-tetrazol-1-
y1)bhenoxylmethyl}-1H-byrazole-5-carbonitrile
reN\
I N 0
INI
A) tert-Butyl 4-(5-cyano-4-{1-4-(1H-tetrazol-1-yl)bhenoxylmethyl}-1H-byrazol-1-
y1)piperidine-1-carboxylate
To a stirred, cold (0 degrees Celsius) solution of triphenylphosphine (283 mg,
1.08
mmol) in tetrahydrofuran (2 mL) was added diethylazodicarboxylate (0.17 mL,
1.1
mmol) drop wise. The cold reaction mixture was stirred for 20 minutes before a
solution
of 4-tetrazol-1-yl-phenol (165.5 mg, 1.021 mmol) in tetrahydrofuran was added.
After 35
minutes a solution of tert-butyl 4-(5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yl)piperidine-
1-carboxylate (Preparation 15) (300 mg, 0.979 mmol) in tetrahydrofuran was
added and
the reaction was slowly allowed to warm up to room temperature overnight. The
reaction was concentrated under reduced pressure and the residue was purified
by
flash chromatography eluting with a gradient from 0% to 80% ethyl acetate in
heptanes
to give the title compound as a white fluffy solid (304 mg, 68%). 1H NMR (400
MHz,
deuterochloroform) delta ppm 1.46 (s, 9 H) 2.03 (s, 2 H) 2.06 - 2.20 (m, 2 H)
2.90 (br. s.,
2 H) 4.28 (br. s., 2 H) 4.46 - 4.56 (m, 1 H) 5.12 (s, 2 H) 7.10 - 7.18 (m, 2
H) 7.59- 7.66
(m, 2 H) 8.90 (s, 1 H); LCMS (ES) 451.1 (M+1)
B) 1-Piperidin-4-y1-4-{1-4-(1H-tetrazol-1-yl)phenoxylmethyl}-1H-pyrazole-5-
carbonitrile
tert-Butyl 4-(5-cyano-4-{[4-(1H-tetrazol-1-y1)phenoxy]methyll-1H-pyrazol-1-
y1)piperidine-
1-carboxylate (298 mg, 0.663 mmol) was dissolved in dichloromethane (1.6
mL). Trifluoroacetic acid (0.15 mL) was added and the reaction was stirred at
room
temperature under nitrogen for 1.5 hours. The reaction was concentrated and
used as
is in the following step without further purification. LCMS (ES+) 351.1 (M+1)
C) 1-Methylcyclopropyl 4-(5-cyano-4-{[4-(1H-tetrazol-1-yl)phenoxy]methyll-1H-
pyrazol-
1-yl)piperidine-1-carboxylate
1-Piperidin-4-y1-4-{[4-(1H-tetrazol-1-yl)phenoxy]methyll-1H-pyrazole-5-
carbonitrile (30
mg, 0.086 mmol) and diisopropylethylamine (0.12 ml, 0.688 mmol) were dissolved
in
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acetonitrile (2 mL) in a sealed tube. 2-Chloro-5-ethylpyrimidine (0.020mL,
0.2 mmol) was added and the reaction was heated at 120 degrees Celsius for 18
hours
and at room temperature for 36 hours. The reaction mixture was concentrated
under
reduced pressure and the crude material was purified by flash chromatography
eluting
with a gradient from 0% to 70% ethyl acetate in heptanes to give a brown
solid. The
solid was triturated with minimal amounts of ether to give the title compound
as a light
brown solid (3 mg, 8%). LCMS (ES+) 457.1 (M+1) 1H NMR (400 MHz,
deuterochloroform) delta ppm 1.16- 1.22(m, 3 H) 2.10 (br. s., 2 H) 2.13 - 2.25
(m, 2 H)
2.43 - 2.50 (m, 2 H) 3.00 - 3.10 (m, 2 H) 3.43 -3.50 (m, 1 H) 4.88 - 4.96 (m,
2 H) 5.12 (s,
2 H) 7.10 - 7.16 (m, 2 H) 7.60 - 7.64 (m, 2 H) 7.66 (s, 1 H) 8.12 - 8.24 (m, 2
H) 8.90 (s, 1
H)
Example 40: Isopropyl 4-{5-cyano-4-[(3-cyanophenoxy)methy1]-1H-pyrazol-1-
yllpiperidine-1-carboxylate
o)
N\
\ N\
= 0\
--N
The title compound was prepared using commercially available 3-cyanophenol,
following procedures analogous to Example 12. The crude material was purified
by
flash chromatography eluting with a gradient of 0% to 40% ethyl acetate in
heptane to
give 16.4 mg (62%) of the title compound as a clear colorless residue. 1H NMR
(400
MHz, deuterochloroform) delta ppm 1.28(d, J=6.25 Hz, 6 H) 1.99 - 2.09 (m, 2 H)
2.10 -
2.24 (m, 2 H) 2.88 - 3.06 (m, 2 H) 4.35 (br. s., 2 H) 4.48 - 4.60 (m, 1 H)
4.90 - 5.01 (m, 1
H) 5.08 (s, 2 H) 7.19 - 7.25 (m, 2 H) 7.32 (d, J=7.82 Hz, 1 H) 7.39 - 7.47 (m,
1 H) 7.68 (s,
1 H)
Example 41: Isopropyl 4-{5-cyano-4-1-(4-cyano-3-methylphenoxy)methy11-1H-
pyrazol-1-
yllpiperidine-1-carboxylate
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N
lei 00%
\ 0 (
....... /N _______________________________ ( N
----N / 0
The title compound was prepared using 4-hydroxy-2-methylbenzonitrile,
following
procedures analogous to Example 12. The crude material was purified by flash
chromatography eluting with a gradient from 0% to 40% ethyl acetate in
heptanes to
give 18.8 mg (69%) of the title compound as a clear residue. 1H NMR (400 MHz,
deuterochloroform) delta ppm 1.27 (d, J=6.25 Hz, 6 H) 1.96 - 2.08 (m, 2 H)
2.09 - 2.23
(m, 2 H) 2.54 (s,3 H) 2.96 (t, J=12.51 Hz, 2 H) 4.35 (br. s., 2 H) 4.54 (tt,
J=11.29, 4.15
Hz, 1 H) 4.95 (spt, J=6.25 Hz, 1 H) 5.09 (s, 2 H) 6.85 (dd, J=8.60, 2.35 Hz, 1
H) 6.90 (s,
1 H) 7.57 (d, J=8.60 Hz, 1 H) 7.67 (s, 1 H)
Example 42 : Isopropyl 4-{5-cyano-4-[(4-cyanophenoxy)methy1]-1H-pyrazol-1-
yllpiperidine-1-carboxylate
N
Si N
_....:::. /N K i,N1
N 0
The title compound was prepared using commercially available 4-cyanophenol,
following procedures analogous to Example 12. The crude material was purified
by
flash chromatography eluting with a gradient from 0% to 40% ethyl acetate in
heptane
to give 14.7mg (56%) of the title compound as a sticky white solid. 1H NMR
(400 MHz,
deuterochloroform) delta ppm 1.27 (d, J=6.25 Hz, 6 H) 1.95 - 2.08 (m, 2 H)
2.16 (m, 2
H) 2.85 - 3.08 (m, 2 H) 4.35 (br. s., 2 H) 4.54 (tt, J=11.29, 4.15 Hz, 1 H)
4.95 (dt,
J=12.51, 6.25 Hz, 1 H) 5.11 (s, 2 H) 7.04 (d, J=8.99 Hz, 2 H) 7.64 (d, J=8.99
Hz, 2 H)
7.68 (s, 1 H)
Example 43: 4-[(4-Cyano-2-fluorophenoxy)methy1]-111-(5-ethylpyrimidin-2-
y1)piperidin-
4-yll-1 H-pyrazole-5-carbonitri le
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N
\\
40, F
N
? p
----AN-K \--(N-) _____________________________________ /
N \ // \
N
The title compound was prepared using commercially available 4-cyano-2-
fluorophenol,
following procedures analogous to Example 39. The crude material was purified
by
flash chromatography eluting with a gradient from 0% to 1.5% methanol in
dichloromethane. The resulting solids were further purified via
recrystallization from
10% methanol/ethyl acetate to give 3.67 g (60%) of clean product as a nearly
white
solid. 1H NMR (500 MHz, deuterochloroform) delta ppm 1.22 (t, J=7.56 Hz, 3 H)
2.07 -
2.15 (m, 2 H) 2.15 - 2.28 (m, 2 H) 2.50 (q, J=7.56 Hz, 2 H) 3.03 - 3.13 (m, 2
H) 4.66 (tt,
J=11.44, 4.18 Hz, 1 H) 4.95 (d, J=13.66 Hz, 2 H) 5.19 (s, 2 H) 7.13 (t, J=8.17
Hz, 1 H)
7.42 (dd, J=10.37, 1.83 Hz, 1 H) 7.47 (d, J=8.29 Hz, 1 H) 7.70 (s, 1 H) 8.21
(s, 2 H)
Example 44: tert-Butyl 4-{5-cyano-4-[(4-cyano-2-fluorophenoxy)methy1]-1H-
pyrazol-1-
yllpiperidine-1-carboxylate
N
\\
fib F
N
0/
N _______________________________________ ( )N __ . (
1\1/ 0
The title compound was prepared using commercially available 4-cyano-2-
fluorophenol,
following procedures analogous to Example 15. The crude material was purified
by
flash chromatography eluting with a gradient from 10% to 40% ethyl acetate in
heptanes
to give the title compound (21 g, 100%). 1H NMR (deuterochloroform) delta ppm
7.71
(s, 1H), 7.44 - 7.48 (m, 1H), 7.40 - 7.43 (m, 1H), 7.09 - 7.15 (m, 1H), 5.18
(s, 2H), 4.48 -
4.56 (m, 1H), 4.22 - 4.38 (m, 2H), 2.84 - 3.01 (m, 2H), 2.09 - 2.19 (m, 2H),
1.99 - 2.06
(m, 2H), 1.49 (s, 9H)
Example 45: Isopropyl 4-{5-cyano-4-1-(2-cyano-4-fluorophenoxy)methy11-1H-
pyrazol-1-
yllpiperidine-1-carboxylate
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F
. ---N
N
1:::/
N--( \14-(
_______________________________________________ 0
The title compound was prepared using commercially available 2-cyano-4-
fluorophenol,
following procedures analogous to Example 15. The crude material was purified
by
HPLC (Column Waters Atlantis dC18 4.6x5Omm, 5 micrometer; Modifier: 0.05%
trifluoroacetic acid; Gradient: 95% water! 5% acetonitrile linear to 5% water!
95%
acetonitrile over 4.0 min, HOLD at 5% water! 95% acetonitrile to 5.0 min;
Flow: 2.0
mL/min) to give 35.8 mg (73%) of the title compound. LCMS (ES+): 412. 0 (M+1)
Example 46: Isopropyl 4-(5-cyano-4-{1-4-(dimethylcarbamoy1)-2-
fluorophenoxylmethyll-
1H-pyrazol-1-yl)piperidine-1-carboxylate
\
N'
0
ilk F
N
? p
N _______________________________________ ( IN
-------N/
0
The title compound was prepared using 3-fluoro-4-hydroxy-N,N-dimethylbenzamide
(Preparation 31B), following procedures analogous to Example 15. The crude
material
was purified by HPLC (Column Waters Atlantis dC18 4.6x5Omm, 5 micrometer;
Modifier: 0.05% trifluoroacetic acid; Gradient: 95% water! 5% acetonitrile
linear to 5%
water! 95% acetonitrile over 4.0 min, HOLD at 5% water! 95% acetonitrile to
5.0 min;
Flow: 2.0mL/min) to give 6.8 mg (12%) of the title compound. LC/MS (ES+):
458.0
(M+1)
Example 47: 1-Methylcyclopropyl 4-(5-cyano-4-{[4-(dimethylcarbamoy1)-2-
fluorophenoxylmethy11-1H-pyrazol-1-yl)piperidine-1-carboxylate
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\
N.--
0
fjk F
N
0 p
7
--------N,
0
The title compound was prepared using 3-fluoro-4-hydroxy-N,N-dimethylbenzamide
(Preparation 31B), following procedures analogous to Example 15. The crude
material
was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 micrometer;
Modifier: 0.05% trifluoroacetic acid; Gradient: 95% water! 5% acetonitrile
linear to 5%
water! 95% acetonitrile over 4.0 min, HOLD at 5% water! 95% acetonitrile to
5.0 min;
Flow: 2.0mL/min) to give 28.7 mg (51%) of the title compound. LC/MS ( ES+):
470.1
(M+1)
Example 48: 1-Methylcyclopropyl 4-(5-cyano-4-{1-2-fluoro-4-
(methylcarbamoyl)phenoxylmethyly1H-pyrazol-1-yl)piperidine-1-carboxylate
\NH
0
4410 F
N
___________________________________________ \
-- I
N -(N I
0
The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide
(Preparation 31A), following procedures analogous to Example 15. The crude
material
was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 micrometer;
Modifier: 0.05% trifluoroacetic acid; Gradient: 95% water! 5% acetonitrile
linear to 5%
water! 95% acetonitrile over 4.0 min, Hold at 5% water! 95% acetonitrile to
5.0 min;
Flow: 2.0mL/min) to give 35.6 mg (65%) of the title product. LC/MS (ES+):
456.0 (M+1)
Example 49: 4-({5-Cyano-141-(5-ethylpyrimidin-2-yl)piperidin-4-y1F1H-pyrazol-4-
yllmethoxy)-3-fluoro-N,N-dimethylbenzamide
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\----
0
. F
N
U/
....._ ( __________________________________ \ N=-) /
N ,N1- / _____________________________________________
1\1/ _____________________________________ i N
The title compound was prepared using 3-fluoro-4-hydroxy-N,N-dimethylbenzamide
(Preparation 31B), following procedures analogous to Example 39. The crude
material
was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 micrometer;
Modifier: 0.05% trifluoroacetic acid; Gradient: 95% water! 5% acetonitrile
linear to 5%
water! 95% acetonitrile over 4.0 min, HOLD at 5% water! 95% acetonitrile to
5.0 min;
Flow: 2.0mL/min) to give 26.7 mg of the title product. LC/MS (ES+): 478.0
(M+1)
Example 50: 1-Methylcyclopropyl 4-{5-cyano-4-[(4-cyano-2-fluorophenoxy)methy11-
1H-
pyrazol-1-yllpiperidine-1-carboxylate
N
\\
4* F
N
N--( N-
1\1/ ______________________________________ / o
The synthesis is outlined in Scheme 5 below.
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0
o ) ii c.......7H
8 _____________________________________ ( : ___________
Step A N,
N ----":":N
+ r\j--
OH Ns NN Step B
1\1 II---) =N
N-N
0 0 H --.
N 1\1
.......--,..õ,
0 0 0 0
õ....---,.õ, ......--,...õ..
Step C
N
HO $
F
//N
/P di 0 N
0 //
F ii (---0
F
(-0
F
N
µ1\1-...'-'N N \ _.,_._.
'1\1---'----N Step D N ) --..--_,.õ.
Step E
N-----N
-.. ..--
N 1\1
H
0 0 Ts0H
0 0
,.....---,..õ,
Scheme 5
A) tert-Butyl 4-1-5-cyano-4-(ethoxycarbony1)-1H-pyrazol-1-yllpiperidine-1-
carboxylate
N
0 ///
/N--(
Ethyl 5-cyano-1H-pyrazole-4-carboxylate (Jubilant Chemsys Ltd. D-12, Sector-
59, 201
301, Noida, U.P. India) (50 g, 300 mmol), tert-butyl 4-hydroxypiperidine-1-
carboxylate
(67 g, 333 mmol), and triphenylphosphine (111 g, 420 mmol) were dissolved in 2-
methyl
tetrahydrofuran (200 mL) and cooled to 0 degrees Celsius. A 40% solution of
diethyl
azodicarboxylate in toluene (76.5 mL, 420 mmol) was added drop wise. Once the
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addition was complete, the reaction was warmed up to room temperature over 1
hour
and then allowed to stir at room temperature for 18 hours. Under vigorous
stirring,
heptane (1400 mL) was carefully added and a suspension formed after 1 hour.
The
solids were filtered off, and the filtrate was washed with a mixture of
heptane (400 mL)
and ethyl acetate (200 mL). The filtrate was then concentrated and the residue
was
purified by flash chromatography eluting with 25% ethyl acetate in heptanes
and then
re-crystallized from ethyl acetate-heptane to give the desired product (35.2
g, 33%). 1H
NMR (deuterochloroform) delta ppm 7.97 (s, 1H), 4.49 - 4.59 (m, 1H), 4.36 (q,
J = 7.1
Hz, 2H), 4.22 - 4.30 (m, 2H), 2.80 - 2.99 (m, 2H), 2.06 - 2.19 (m, 2H), 1.93 -
2.02 (m,
2H), 1.46 (s, 9H), 1.37 (t, J = 7.1 Hz, 3H)
B): tert-Butyl 4-1-5-cyano-4-(hydroxymethyl)-1H-byrazol-1-yllbiberidine-1-
carboxylate
N
oi-(_...,/
_______________________________________ \ o (
7
o
tert-Butyl 4[5-cyano-4-(ethoxycarbony1)-1H-pyrazol-1-yl]piperidine-1-
carboxylate (45.5
g, 131 mmol) was dissolved in tetrahydrofuran (350 mL) and cooled to -78
degrees
Celsius. A 1.5M solution of diisobutylaluminum hydride in toluene (50 g, 350
mmol) was
added drop wise over 75 minutes maintaining the internal temperature between -
65
degrees Celsius and -60 degrees Celsius. Once the addition was complete, the
reaction mixture was warmed to -10 degrees Celsius for 90 minutes. While
maintaining
a temperature of -10 degrees Celsius, an aqueous 4 M solution of potassium
hydroxide
(350 mL, 10.7 eq) was carefully added drop wise. Once addition was complete,
the
reaction mixture was slowly allowed to come up to room temperature with
vigorous
stirring and then allowed to stir at room temperature for 20 hours. Methyl
tert-butyl
ether (200 mL) and heptanes (400 mL) were added and the organic phase was
separated. The organic phase was washed with 1 M aqueous potassium hydrogen
sulfate, brine, and dried over a mixture of magnesium sulfate and 30 g of
silica gel. The
solids were filtered off and the filtrate was concentrated under reduced
pressure. Upon
concentration a precipitate began to form. The resulting wet residue was
triturated with
500 mL of 10% methyl tert-butyl ether in heptane at 60 degrees Celsius for 1
hour and
the suspension was slowly cooled to room temperature under stirring. The
resulting
solids were filtered off and dried in a vacuum oven set at 40 degrees Celsius
to give
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tert-butyl 4[5-cyano-4-(hydroxymethyl)-1H-pyrazol-1-yl]piperidine-1-
carboxylate (31.2g,
78%). 1H NMR (deuterochloroform) delta ppm 7.59 (s, 1H), 4.70 (d, J = 5.5 Hz,
2H),
4.41 -4.51 (m, 1H), 4.17 - 4.32 (m, 2H), 2.81 -2.96 (m, 2H), 2.01 -2.16 (m,
3H), 1.94 -
2.00 (m, 2H), 1.45 (s, 9H)
C) tert-Butyl 4-{5-cyano-4-1-(4-cyano-2-fluorophenoxy)methy11-1H-pyrazol-1-
yllpiperidine-1-carboxylate
N
\\
OF
N
? 9/
. 0 (
CN-( 1\N
-----NI
0
To a 4 L bottle was charged tert-butyl 445-cyano-4-(hydroxymethyl)-1H-pyrazol-
1-
yl]piperidine-1-carboxylate (336 g, 1.10 moles), triphenylphosphine (359.58 g,
1.37
moles), 4-cyano-2-fluorophenol (157.89 g, 1.15 moles), and 2-
methyltetrahydrofuran
(2.02 L, 20.10 moles). The mixture was stirred into solution and kept under
nitrogen at
room temperature. To another 4 L bottle was charged, a solution of diethyl
diazenedicarboxylate in toluene (564.33 mL, 620.76 g, 1.43 moles) and 2-
methyltetrahydrofuran (2.12 L, 21.11 moles). The mixture was agitated to
ensure
complete solution, and kept under nitrogen at room temperature. A single
peristaltic
pump was used (two feed lines) to pump the two streams to a T-piece
(stainless)
followed by 100 mL of coil volume (1/8" followed by 1/4" ID PTE tubing) with a
combined flow rate of 20 mL/min. After 8 hours of flowing, the feed bottles
were
emptied, and 2 x 25 mL of methyltetrahydrofuran was used to rinse the bottles,
and
pumped through the lines. The product stream was used as is in the following
reaction.
D) 4-1-(4-Cyano-2-fluorophenoxy)methy11-1-piperidin-4-y1-1H-pyrazole-5-
carbonitrile
tosylate salt
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F
N( .\NH Ts0H
The stream of tert-butyl 4-{5-cyano-4-[(4-cyano-2-fluorophenoxy)methy1]-1H-
pyrazol-1-
yllpiperidine-1-carboxylate collected in Step C was split into two 5 L single
neck
flasks. p-Toluenesulfonic acid monohydrate (344.22 g, 1.81 moles) was split
charged
into the mixture and the flask was heated using a rotary evaporator bath kept
at 75
degrees Celsius for 8 hours. The reaction was allowed to cool to room
temperature and
granulated overnight. The mixture was filtered and pulled dry under vacuum for
3 hours
to give the target product as the tosylate salt (480 g, 88% over two steps).
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E) 1-Methylcyclopropyl 4-{5-cyano-4-[(4-cyano-2-fluorophenoxy)methy1]-1H-
pyrazol-1-
yllpiperidine-1-carboxylate
N
\\
OF
N
U/
N ,N
--:N/ < _____________________________________ / 0
The tosylate salt of 4-[(4-cyano-2-fluorophenoxy)methy1]-1-piperidin-4-y1-1H-
pyrazole-5-
carbonitrile (478 g, 960.71 mmoles) was dissolved in 2-methyltetrahydrofuran
(2.39 L,
23.83 moles) and water (478.00 mL) in a 4L bottle. Triethylamine (200.86 mL,
1.44
moles) and 1-methylcyclopropyl 4-nitrophenyl carbonate (Preparation 26)
(229.83 g,
960.71 mmoles) were added and stirred for 48 hours. The reaction mixture was
washed
with aqueous 1N sodium hydroxide (1 L). The mixture was stirred and the layers
separated. The organic layer was washed several times with aqueous 1N sodium
hydroxide (1 L), dried over magnesium sulfate, filtered and the filtrate was
concentrated
under reduced pressure to give bright yellow solids. These solids were
slurried in ethyl
acetate at room temperature overnight. The solids were filtered and the
resulting pale
yellow solids were re-slurried in ethyl acetate (3 volumes), filtered and
pulled dry under
vacuum to give the target compound as an off-white solids (313 g, in two
batches, 77%).
1H NMR (400 MHz, deuterochloroform) delta ppm 0.60 - 0.66 (m, 2 H) 0.84 - 0.90
(m, 2
H) 1.55(s, 3 H) 1.96 - 2.04 (m, 2 H) 2.11 (qd, J=12.10, 4.68 Hz, 2 H) 2.92
(br. s., 2 H)
4.07 - 4.41 (m, 2 H) 4.50 (tt, J=11.27, 4.15 Hz, 1 H) 5.16 (s, 2 H) 7.09 (t,
J=8.20 Hz, 1
H) 7.36 - 7.46 (m, 2 H) 7.68 (s, 1 H).
Melting point = 144.6 degrees Celsius
Combustion Analysis for (Quantitative Technologies Inc. (QTI)
291 Route 22 East
Salem Ind. Park - Bldg 5
Whitehouse NJ 08888-0470
C22H22FN503
C (Theoretical=62.40%)
62.28 %
62.29 %
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H (Theoretical=5.24%)
5.17 %
5.13 %
N (Theoretical=16.54%)
16.42%
16.50 %
Example 51: tert-Butyl (3S,4S)-4-(5-cyano-4-{1-2-fluoro-4-
(methylcarbamoyl)ohenoxylmethyll-1H-byrazol-1-y1)-3-fluorobioeridine-1-
carboxylate
\NH
0 41 F
N
Occt F
-- N...... \N __ .( (
N -/ \O
A) tert-Butyl (3S,4S)-4-[5-cyano-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-
fluoropiperidine-
1-carboxylate
tert-Butyl (3S,4S)-4-[5-cyano-4-(ethoxycarbony1)-1H-pyrazol-1-y1]-3-
fluoropiperidine-1-
carboxylate was prepared from ethyl 5-cyano-1H-pyrazole-4-carboxylate and
(3S,4R)-
tert-butyl 3-fluoro-4-hydroxypiperidine-1-carboxylate (Preparation 43 B) in a
manner
similar to that described for the preparation of tert-butyl 445-cyano-4-
(ethoxycarbony1)-
1H-pyrazol-1-yl]piperidine-1-carboxylate (Example 50, Step A). The crude
material was
purified by flash chromatography eluting with a gradient from 0% to 30% ethyl
acetate in
heptanes to give the desired product as a thick clear oil, (149.4 mg, 32%).
B) tert-Butyl (35,45)-415-cyano-4-(hydroxymethyl)-1H-pyrazol-1-y1]-3-
fluoropiperidine-
1-carboxylate
tert-Butyl (35,45)-445-cyano-4-(hydroxymethyl)-1H-pyrazol-1-y1]-3-
fluoropiperidine-1-
carboxylate was prepared from tert-butyl (3S,4S)-4-[5-cyano-4-(ethoxycarbony1)-
1H-
pyrazol-1-y1]-3-fluoropiperidine-1-carboxylate in a manner similar to that
described for
the preparation of tert-butyl 445-cyano-4-(hydroxymethyl)-1H-pyrazol-1-
yl]piperidine-1-
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carboxylate (Example 50, Step B). The crude product was purified by flash
chromatography eluting with a gradient from 5% to 50% ethyl acetate in
heptanes to
give the desired product as a thick clear oil that solidified upon standing
(74 mg, 56%).
C) tert-Butyl (35,45)-4-(5-cyano-4-{1-2-fluoro-4-
(methylcarbamoyl)phenoxylmethy11-1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate
The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide
(Preparation 31A), following procedures analogous to Example 50. The crude
material
was purified by HPLC (Column: Waters Xbridge C12 4.6x5Omm, 5 micrometer;
Modifier:
0.05% Ammonium hydroxide; Gradient: 95% water! 5% acetonitrile linear to 5%
water!
95% acetonitrile over 4.0 min, HOLD at 5% water! 95% acetonitrile to 5.0 min;
Flow:
2.0mL/min) to give the desired product. LC/MS (ES+): 476.4 (M+1)
Example: 52: tert-Butyl (3R,45)-4-(5-cyano-4-{[2-fluoro-4-
(methylcarbamoyl)phenoxy]methy11-1H-pyrazol-1-y1)-3-fluoropiperidine-1-
carboxylate
\NH
0 =:N
y/
/ / E
-- \ 0<
--- N-..< /N
N 0
The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide
(Preparation 31A), following procedures analogous to Example 51. The crude
material
was purified by HPLC (Column: Waters Xbridge C12 4.6x50 mm, 5 micrometer;
Modifier: 0.05% Ammonium hydroxide; Gradient: 95% water! 5% acetonitrile
linear to
5% water! 95% acetonitrile over 4.0 min, HOLD at 5% water! 95% acetonitrile to
5.0
min; Flow: 2.0mL/min) to give the desired product. LC/MS (ES+): 476.4 (M+1)
Example 53: 1-Methylcyclopropyl (35,45)-4-(5-cyano-4-{[2-fluoro-4-
(methylcarbamoyl)phenoxylmethy11-1H-pyrazol-1-y1)-3-fluoropiperidine-1-
carboxylate
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\NH
0 F
yt F
________________________________________________ 0
The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide
(Preparation 31A), following procedures analogous to Examples 50 and 51. The
crude
material was purified via HPLC (Column: Princeton 2-ethyl pyridine 250 x 21.2
mm 5
micrometer; Gradient: 95% heptane / 5% ethanol for 1.5 minutes, linear to 0%
heptane
/100% ethanol over 10 min, HOLD at 0% heptane /100% ethanol to 5.0 minfor 1
minute and linear to 95% heptane / 5% ethanol ; Flow: 28 mL/min) to give the
desired
product. LC/MS (ES+): 473.9 (M+1)
Example 54: 1-Methylcyclopropyl (3R,4R)-4-(5-cyano-4-{[2-fluoro-4-
(methylcarbamoyl)phenoxy1methy11-1H-pyrazol-1-y1)-3-fluoropiperidine-1-
carboxylate
\N H
0 F
0/
/N--(/ 0
The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide
(Preparation 31A), following procedures analogous to Examples 50 and 51. The
crude
material was purified via HPLC (Column: Princeton 2-ethyl pyridine 250 x
21.2mm, 5
micrometer; Gradient: 95% heptane / 5% ethanol for 1.5 minutes, linear to 0%
heptane
/ 100% ethanol over 10min, Hold at 0% heptane / 100% ethanol to 5.0 minfor 1
minute
and linear to 95% heptane / 5% ethanol; Flow: 28 mL/min) to give the desired
product.
LC/MS (ES+): 473.9 (M+1)
Example 55: tert-Butyl (3S,4S)-4-(5-cyano-4-{[(2-methylpyridin-3-
yl)oxylmethyll-1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate
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, 1 N
.z.-----N _______________________________ / 0
tert-Butyl 4-(5-cyano-4-{[(methylsulfonyl)oxy]methyll-1H-pyrazol-1-y1)-3-
fluoropiperidine-
1-carboxylate (Preparation 42) (33 mg, 0.082 mmol) was dissolved in
acetonitrile (3 mL)
and cesium carbonate (53 mg, 0.164mmol) and 3-hydroxy-2-methylpyridine (9 mg,
0.082 mmol) were added. The reaction mixture was heated to 80 degrees Celsius
for
1.5 hour. The reaction was cooled to room temperature and concentrated under
reduced pressure. The crude residue was diluted with water and extracted with
ethyl
acetate (3x). The combined organic extracts were washed with aqueous 0.5N
sodium
hydroxide, water and brine and dried over sodium sulfate, filtered and the
filtrate was
concentrated under reduced pressure. The crude residue was purified by flash
chromatography eluting with a gradient from 30% to 100% ethyl acetate in
heptanes to
give the racemic product as an amber oil (30 mg, 70%). 1H NMR (500 MHz,
deuterochloroform) delta ppm 1.50 (s, 9 H) 2.11 (m, 1 H) 2.25 -2.39 (m, 1 H)
2.53 (s, 3
H) 2.93 (br. s., 2 H) 4.30 (br. s., 1 H) 4.43 - 4.71 (m, 2 H) 4.72 - 4.91 (m,
1 H) 5.09 (s, 2
H) 7.09 - 7.20 (m, 2 H) 7.75 (s, 1 H) 8.16 (d, J=3.90 Hz, 1 H)
Example 56: tert-Butyl (3S,4R)-4-(5-cyano-4-{[(2-methylpyridin-3-
yl)oxylmethyll-1H-
pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate
N
1
//N/I F
--c--
N-- \N-i (
/ 0
The title compound was prepared using commercially available 3-hydroxy-2-
methylpyridine, following procedures analogous to Example 55. The crude
material was
purified by flash chromatography eluting with a gradient from 30% to 100%
ethyl acetate
in heptane to give the desired product as an amber oil. 1H NMR (500 MHz,
deuterochloroform) delta ppm 1.50 (s, 9H), 2.01 - 2.08 (m, 1 H) 2.52 (s, 3 H)
2.74 - 2.88
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(M, 1 H) 2.94 - 3.14 (m, 1 H) 3.14 - 3.34 (m, 1 H) 4.27 - 4.57 (m, 2 H) 4.61 -
4.75 (m, 1
H) 4.80 - 5.01 (m, 1 H) 5.09 (s, 2 H) 7.10 - 7.18 (m, 2 H) 7.73 (s, 1 H) 8.15
(d, J=3.66 Hz,
1 H)
Example 57: 1-Methylcyclopropyl (3S,4R)-4-(5-cyano-4-{[(2-methylpyridin-3-
yl)oxylmethyll-1H-pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate
1 N
-10// F
- \ 0-.
IN ,N1
N --- ____________________________________ / 0
The title compound was prepared using commercially available 3-hydroxy-2-
methylpyridine, following procedures analogous to Example 55. The crude
material was
purified by flash chromatography, eluting with a gradient of 40% to 100% ethyl
acetate
in heptanes to give the desired racemic product as a white solid. 1H NMR (400
MHz,
deuterochloroform) delta ppm 0.66 (br. s., 2 H) 0.91 (br. s., 2 H) 1.57 (s, 3
H) 2.05 (d,
J=12.10 Hz, 1 H) 2.52 (s, 3 H) 2.82 (br. d, J=9.00 Hz, 1 H) 3.05 (br. d,
J=9.00 Hz, 1 H)
3.15 - 3.40 (m, 1 H) 4.20 - 4.60 (m, 2 H) 4.60 - 4.77 (m, 1 H) 4.77 - 5.03 (m,
1 H) 5.09 (s,
2 H) 7.06 - 7.21 (m, 2 H) 7.73 (s, 1 H) 8.16 (d, J=4.29 Hz, 1 H)
Example 58: 1-Methylcyclopropyl (3S,4R)-4-(5-cyano-4-{[(2-methylpyridin-3-
yl)oxylmethyll-1H-pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate
, I
CI:1 97 F
i"--c
N-- \N 1
/ "a \1/
The title compound was prepared using commercially available 3-hydroxy-2-
methylpyridine, following procedures analogous to Example 55. The crude
material was
purified by flash chromatography eluting with a gradient of 40% to 100% ethyl
acetate in
heptanes to give the racemic product which was further purified by chiral HPLC
with the
following conditions: Column: chiralcel OJ-H 4.6mm x 25cm; Mobile Phase: 85/15
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carbon dioxide/methanol, Modifier: 0.2% isopropylamine; Flow Rate:
2.5mL/minute to
give the title compound. LC/MS (ES+): 414.1 (M+1)
Example 59: 1-Methylcyclopropyl (3R,4S)-4-(5-cyano-4-{[(2-methylpyridin-3-
yl)oxy]methy11-1H-pyrazol-1-y1)-3-fluoropiperidine-1-carboxylate
, I
9% F
'-:
, 0
----CNI......( \N ______________________________ ¨.
/ o
The title compound was prepared using commercially available 3-hydroxy-2-
methylpyridine, following procedures analogous to Example 55. The crude
material was
purified by flash chromatography eluting with a gradient from 40% to 100%
ethyl acetate
in heptanes to give the racemic product which was further purified by chiral
HPLC with
the following conditions: Column: chiralcel OJ-H 4.6mm x 25cm; Mobile Phase:
85/15
carbon dioxide/methanol, Modifier: 0.2% isopropylamine; Flow Rate:
2.5mL/minute to
give the title compound. LC/MS (ES+): 414.1 (M+1)
Example 60: tert-Butyl 4-(5-cyano-4-{[4-(1H-1,2,3-triazol-1-yl)phenoxy]methyll-
1H-
pyrazol-1-yl)piperidine-1-carboxylate
N/
NN
41 N
? p
. ______________________________________________ 0 (----CN-( /\N
------N/
0
The title compound was prepared using 4-(1H-1,2,3-triazol-1-yl)phenol (US
Patent
Application No. PCT/U52009/038315, Publication No. WO 2009/129036 Al)
following
procedures analogous to Example 15. The crude material was purified by HPLC
(Column: Phenomenex Gemini C18 250x21.2 mm, 8 micrometer; Mobile Phase: from
50% acetonitrile (ammonia pH 10) in water (ammonia pH 10) to 55% acetonitrile
(ammonia pH 10) in water (ammonia pH 10); Flow Rate: 25mUminute; wavelength:
220
nm) to give the title compound. LC/MS (ES+): 450.1 (M+1)
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Example 61: tert-Butyl 4-(5-cyano-4-{[4-(2H-1,2,3-triazol-2-yl)phenoxy]methyll-
1H-
pyrazol-1-yl)piperidine-1-carboxylate
Op
N--11
'N
? p
N __________________________________________ / __ (
0
The title compound was prepared using 4-(2H-1,2,3-triazol-2-yl)phenol (US
Patent
Application No. PCT/U52009/038315, Publication No. WO 2009/129036 Al)
following
procedures analogous to Example 15. The crude material was purified by HPLC
(Column: Phenomenex Gemini C18 250x21.2 mm, 8 micrometer; Mobile Phase: 63%
acetonitrile (ammonia pH 10) in water (ammonia pH 10); Flow Rate: 25mUminute;
wavelength: 220 nm) to give the title compound. LC/MS (ES+): 450.1 (M+1)
Example 62: 1-Methylcyclopropyl 4-(4-((4-(1H-1,2,3-triazol-1-
y1)phenoxy)methyl)-5-
cyano-1H-pyrazol-1-ypoiperidine-1-carboxylate
,N
N '
N---1
#
N ,
N --- N
)\
N
00
1>'
The title compound was prepared in a manner analogous to Example 60 starting
with
Example 60. The crude material was purified by reverse phase HPLC:
Column: Kromasil Eternity-5-C18 150x30 mmx 5 micrometer
Mobile phase: from 38% acetonitrile (0.225% formic acid) in water (0.225%
formic acid)
to 58% acetonitrile (0.225% formic acid) in water (0.225% formic acid)
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Flow rate: 30 mL/min
Wavelength: 220 nm
1H NMR (400 MHz, deuterochlorform): delta ppm 7.92 (s, 1H), 7.84 (s, 1H), 7.68
(d,
3H), 7.11 (t, 2H), 5.11 (s, 2H), 4.53 (m, 1H), 4.26 (m, 2H), 2.93 (s, 2H),
2.12(t, 2H), 2.03
(d, 2H) 1.56 (s, 3H), 0.88 (t, 2H), 0.64 (t, 2H)
Example 63: 1-Methylcyclopropyl 4-(4-((4-(2H-1,2,3-triazol-2-
yl)phenoxy)methyl)-5-
cyano-1H-pyrazol-1-yl)piperidine-1-carboxylate
N)
1%\i"--N
1110
s 0
c.......
N ,
N ----N
a
N
00
The title compound was prepared in a manner analogous to Example 61 starting
with
Example 61. The crude residue was purified by preparative HPLC to yield 50 mg
(39%)
of the title compound as a white solid:
Column: Boston Symmetrix ODS-H 150x30 mm x 5 micrometer
Mobile phase: from 50% acetonitrile (0.225% formic acid) in water (0.225%
formic acid)
to 70% acetonitrile (0.225% formic acid) in water (0.225% formic acid)
Flow rate: 30 mL/min
Wavelength: 220 nm
1H NMR (400 MHz, deuterochloroform): delta ppm 8.01 (d, 2H), 7.78 (s, 2H),
7.69 (s,
1H), 7.07 (d, 2H), 5.10 (s, 2H), 4.51 (m, 1H), 4.33 (m, 2H), 2.93 (s, 2H),
2.11 (t, 2H),
2.03 (d, 2H) 1.56 (s, 3H), 0.80 (s, 2H), 0.65 (d, 2H).
Example 64: tert-Butyl 4-1-5-cyano-4-(ff1-(methylsulfonyl)piperidin-4-
ylloxylmethyl)-1H-
pyrazol-1-yllpiperidine-1-carboxylate
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, 0---/
N# c_.....õ
,
N ---N
---
N
0 0
/\
The title compound was prepared in a manner analogous to Example 13. The crude
compound was purified by silica gel chromatography using an 1:4 mixture of
petroleum
ether and ethyl acetate.
1H NMR (400 MHz, deuterochloroform): delta ppm 7.54 (s, 1H), 4.53 (s, 2H),
4.48 (m,
1H), 4.28 (br, 2H), 3.69 (m, 1H), 3.31 (m, 4H), 2.90 (m, 2H), 2.79 (s, 3H),
2.11(m, 2H),
1.88-2.00 (m, 6H), 1.47 (s, 9H).
Example 65: tert-Butyl 4-1-5-cyano-4-({2-fluoro-4-[(2-
hydroxyethyl)(methyl)carbamoyllphenoxylmethyl)-1H-pyrazol-1-yllpiperidine-1-
carboxylate
0
N/",,,7OH
s o10 \
F
N ,
N --N
..---
N
00
õ.õ---......,
The title compound was prepared in a manner analogous to Example 46. The crude
material was purified by reverse phase HPLC:
Column: Phenomenex Gemini C18 250x21.2 mm x 8 micrometer
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Mobile phase: from 40% acetonitrile (ammonia pH 10) in water (ammonia pH 10)
to
60% acetonitrile (ammonia pH 10) in water (ammonia pH 10)
Flow rate: 25 mL/min
Wavelength: 220 nm
1H NMR (400 MHz, deuterochloroform): delta ppm 7.69 (s, 1H), 7.28 (d, 1H),
7.24 (s,
1H), 7.05 (d, 1H), 5.18 (s, 2H), 4.50 (q, 1H) , 4.29 (d, 2H) , 3.90 (s, 2H),
3.71 (s, 2H),
3.10 (s, 3H), 2.91 (s, 2H), 2.14 (q, 2H), 1.99 (s, 2H), 1.48 (s, 9H).
Example 66: tert-Butyl 415-cyano-4-({2-fluoro-4-[(3-hydroxypyrrolidin-1-
yl)carbonyllohenoxylmethyl)-1H-pyrazol-1-yllpiperidine-1-carboxylate
0
sio N\r
OH
c0
F
N ,
N ----N
0 0
/.\
The title compound was prepared in a manner analogous to Example 46. The crude
material was purified by reverse phase HPLC:
Column: Phenomenex Gemini C18 250x21.2 mm x 8 micrometer
Mobile phase: from 40% acetonitrile (ammonia pH 10) in water (ammonia pH 10)
to
60% acetonitrile (ammonia pH 10) in water (ammonia pH 10)
Flow rate: 25 mL/min
Wavelength: 220 nm
1H NMR (400 MHz, deuterochloroform): delta ppm 7.69 (s, 1H), 7.32 (d, 2H),
7.05 (t,
1H), 5.14 (s, 2H), 4.52 (q, 2H) , 4.29 (s, 2H) , 3.78 (d, 2H), 3.64 (d, 1H),
3.45 (d, 1H),
2.90 (s, 2H), 2.14 (q, 2H), 2.00 (d, 4H), 1.47 (s, 9H).
Example 67: tert-Butyl 4-(4-{[4-(azetidin-1-ylcarbonyI)-2-
fluorophenoxy]rnethyll-5-cyano-
1H-pyrazol-1-yl)piperidine-1-carboxylate
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0
110
ii, c_______O
F
N ,
N ---N
N
00
........---,õ
The title compound was prepared in a manner analogous to Example 46. The crude
material was purified by reverse phase HPLC:
Column: Phenomenex Gemini C18 250x21.2 mm x 8 micrometer
Mobile phase: from 40% acetonitrile (ammonia pH 10) in water (ammonia pH 10)
to
60% acetonitrile (ammonia pH 10) in water (ammonia pH 10)
Flow rate: 25 mL/min
Wavelength: 220 nm
1H NMR (400 MHz, deuterochloroform): delta ppm 7.69 (s, 1H), 7.43 (d, 1H),
7.41 (s,
1H), 7.04 (t, 1H), 5.14 (s, 2H), 4.50 (q, 1H) , 4.31 (d, 6H) , 2.90 (d, 2H),
2.38 (q, 2H),
2.15 (q, 2H), 2.08 (d, 2H), 1.47 (s, 9H).
Example 68: 1-Methylcyclopropyl 415-cyano-4-({[1-(methylsulfonyl)piperidin-4-
ylloxylmethyl)-1H-pyrazol-1-yllpiperidine-1-carboxylate
s"
;N,
ri\c '0
ii, c.........07
N ,
N --N
N
00
The title compound was prepared in a manner analogous to Example 64. The crude
material was purified by reverse phase HPLC:
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Column: Phenomenex Synergi C18 150x30 mm x 4 micrometer
Mobile phase: 43% acetonitrile (0.225% formic acid) in water (0.225% formic
acid) to
53% acetonitrile (0.225% formic acid) in water (0.225% formic acid)
Flow Rate: 30mUmin
1H NMR (400 MHz, deuterochloroform): delta ppm 7.54 (s, 1H), 4.53 (s, 2H),
4.48 (m,
1H), 4.35 (d, 2H), 3.69 (m, 1H), 3.31 (m, 4H), 2.91 (m, 2H), 2.78 (s, 3H),
2.12(m, 2H),
1.87-1.98 (m, 6H), 1.56 (s, 3H), 0.88 (t, 2H), 0.66 (t, 2H).
Throughout this application, various publications are referenced. The
disclosures of
these publications in their entireties are hereby incorporated by reference
into this
application for all purposes.
It will be apparent to those skilled in the art that various modifications and
variations can be made in the present invention without departing from the
scope or
spirit of the invention. Other embodiments of the invention will be apparent
to those
skilled in the art from consideration of the specification and practice of the
invention
disclosed herein. It is intended that the specification and examples be
considered as
exemplary only, with a true scope and spirit of the invention being indicated
by the
following claims.
131
