KR20130083915A - 4-(5-cyano-pyrazol-1-yl)-piperidine derivatives as gpr119 modulators - Google Patents

Abstract

Described herein are compounds that modulate the activity of G-protein-coupled receptor GPR119 and their use in the treatment of diseases associated with the modulation of G-protein-coupled receptor GPR119 in animals.

Description

4- (5-Cyano-pyrazol-1-yl) -piperidine derivative as GPR119 modulator {4- (5-CYANO-PYRAZOL-1-YL) -PIPERIDINE DERIVATIVES AS GPR119 MODULATORS}

The present invention relates to a new class of cyanopyrazoles, pharmaceutical compositions containing these compounds, and their use for modulating the activity of the G-protein-coupled receptor GPR119.

Diabetes is a disorder in which high levels of blood sugar occur as a result of abnormal glucose homeostasis. The most common forms of diabetes are type I diabetes (also referred to as insulin-dependent diabetes) and type II diabetes (also referred to as non-insulin-dependent diabetes). Type II diabetes, which accounts for approximately 90% of all diabetes cases, causes microvascular complications (including retinopathy, neuropathy and nephropathy), as well as macrovascular complications (including accelerated atherosclerosis, coronary heart disease and stroke). Is a serious progressive disease.

At present, there is no cure for diabetes. Standard treatments for the disease are limited, with a focus on controlling blood glucose levels to minimize or delay complications. Current treatments target insulin resistance (metformin, thiazolidinedione) or insulin release from beta cells (sulfonylurea, exanatid). Sulfonylureas and other compounds that act through depolarization of beta cells promote hypoglycemia because they stimulate insulin secretion regardless of circulating glucose concentration. One approved drug exenatide stimulates insulin secretion only in the presence of high glucose, but should be injected because of lack of oral bioavailability. Dipeptidyl peptidase IV inhibitor cytagliptin is a novel drug that can increase blood levels of incretin hormones to increase insulin secretion, reduce glucagon secretion and have other less characterized effects. However, cytagliptin and other dipeptidyl peptidase IV inhibitors may also affect the tissue levels of other hormones and peptides, but the long-term consequences of this broader effect have not been fully investigated.

In type II diabetes, muscle, fat and liver cells do not respond normally to insulin. This condition (insulin resistance) may be due to a reduced number of cellular insulin receptors, disruption of cellular signaling pathways, or both. First, beta cells compensate for insulin resistance by increasing insulin release. In the end, however, beta cells are unable to produce enough insulin to maintain normal glucose levels (normal blood glucose), indicating progression to type II diabetes.

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) an increase in basal insulin release observed in type II diabetes as well as an insulin resistant diabetes prediabetes stage of obesity (occurs at low non-stimulatory glucose concentrations), and 2) Failure to increase insulin release above the already elevated basal level, in response to hyperglycemia induction, which does not occur in the prediabetes stage and can signal a transition from a normal-glycemic insulin resistant state to type II diabetes. Current therapies for treating the latter aspects include the administration of beta-cell ATP-sensitive potassium channels that trigger the release of endogenous insulin storage, and the administration of exogenous insulin. Neither has correctly normalized blood glucose levels, both of which carry a risk of causing hypoglycemia.

Therefore, there is a great interest in finding agents that act in a glucose-dependent manner. Physiological signaling pathways that work in this manner, including the gut peptides GLP-1 and GIP, are well known. These hormones transmit signals through cognate G-protein coupled receptors to stimulate the production of cAMP in pancreatic beta-cells. It is clear that the increase in cAMP does not cause stimulation of insulin release during fasting or preprandial conditions. However, the biochemical targets of many cAMPs, including ATP-sensitive potassium channels, voltage-sensitive potassium channels, and extracellular flux machinery, are regulated to significantly increase insulin secretion due to postprandial glucose stimulation. Thus, agonist modulators of novel similarly acting beta-cell GPCRs, such as GPR119, will also stimulate the release of endogenous insulin and promote normalization of glucose levels in patients with type II diabetes. In addition, for example, an increase in cAMP as a result of GLP-1 stimulation promotes beta-cell proliferation, inhibits beta-cell death, thereby improving islet mass. This positive effect on beta-cell mass should be beneficial for type II diabetes, where insufficient insulin is produced.

Metabolic disorders have a negative effect on other physiological systems, often co-occurrence of multiple disease states (eg, type I diabetes, type II diabetes, insufficient glucose tolerance, insulin resistance, hyperglycemia, Hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, dyslipidemia, obesity or cardiovascular disease) or secondary diseases that occur secondary to diabetes, such as kidney disease and peripheral neuropathy. Thus, treatment of diabetic conditions should be beneficial for such correlated disease states.

Summary of the Invention

In accordance with the present invention, a new class of GPR119 modulators has been discovered. These compounds include:

Isopropyl 4- {5-cyano-4-[(2,4-difluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;

Isopropyl 4- {5-cyano-4-[(2-methylphenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;

1-methylcyclopropyl 4- {5-cyano-4-[(2,5-difluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;

1-methylcyclopropyl 4- {5-cyano-4-[(2,3-difluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;

1-methylcyclopropyl 4- {4-[(4-carbamoyl-2-fluorophenoxy) methyl] -5-cyano-1H-pyrazol-1-yl} piperidine-1-carboxyl Rate;

1-methylcyclopropyl 4- {4-[(4-carbamoylphenoxy) methyl] -5-cyano-1H-pyrazol-1-yl} piperidine-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- I) 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-pyra Zol-1-yl) piperidine-1-carboxylate;

Isopropyl 4- (5-cyano-4-((2-fluoro-4- (1- (2-hydroxyethyl) -1H-tetrazol-5-yl) phenoxy) methyl) -1H-pyra Zol-1-yl) piperidine-1-carboxylate;

1-methylcyclopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenoxy] methyl} -1H-pyrazole-1 -Yl) piperidine-1-carboxylate;

1-methylcyclopropyl 4- (5-cyano-4-{[4- (1-methyl-1H-tetrazol-5-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperi Dine-1-carboxylate;

1-Methylcyclopropyl 4- (4-((4-carbamoyl-3-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine-1-carboxyl Rate;

Isopropyl 4- (5-cyano-4- {1- [2-fluoro-4- (methylsulfonyl) phenoxy] ethyl} -1H-pyrazol-1-yl) piperidine-1-car Carboxylates;

Isopropyl 4- (5-cyano-4- {1-[(2-methylpyridin-3-yl) oxy] ethyl} -1 H-pyrazol-1-yl) piperidine-1-carboxylate;

Isopropyl 4- (5-cyano-4- {2- [2-fluoro-4- (methylsulfonyl) phenyl] propyl} -1H-pyrazol-1-yl) piperidine-1-carboxyl Rate;

1-methylcyclopropyl 4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) piperidine-1-carboxylate ;

1-methylcyclopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate ;

Isopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;

Isopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-imidazol-2-yl) phenoxy] methyl} -1H-pyrazol-1-yl) Piperidine-1-carboxylate;

Isopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-imidazol-5-yl) phenoxy] methyl} -1H-pyrazol-1-yl) Piperidine-1-carboxylate;

Isopropyl 4- [5-cyano-4-({[2-methyl-6- (1H-1,2,4-triazol-1-yl) pyridin-3-yl] oxy} methyl) -1H- Pyrazol-1-yl] piperidine-1-carboxylate;

Isopropyl 4- [5-cyano-4-({[2-methyl-6- (1H-1,2,4-triazol-1-yl) pyridin-3-yl] amino} methyl) -1H- Pyrazol-1-yl] piperidine-1-carboxylate;

Isopropyl 4- [5-cyano-4-({[2-methyl-6- (methylsulfonyl) pyridin-3-yl] amino} methyl) -1H-pyrazol-1-yl] piperidine- 1-carboxylate;

1-methylcyclopropyl 4- (5-cyano-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine-1- Carboxylates;

1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyra Sol-5-carbonitrile;

Isopropyl 4- {5-cyano-4-[(3-cyanophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;

Isopropyl 4- {5-cyano-4-[(4-cyano-3-methylphenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate;

Isopropyl 4- {5-cyano-4-[(4-cyanophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;

4-[(4-cyano-2-fluorophenoxy) methyl] -1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -1H-pyrazole-5 Carbonitrile;

tert-butyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate;

Isopropyl 4- {5-cyano-4-[(2-cyano-4-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate;

Isopropyl 4- (5-cyano-4-{[4- (dimethylcarbamoyl) -2-fluorophenoxy] methyl} -1 H-pyrazol-1-yl) piperidine-1-carboxyl Rate;

1-methylcyclopropyl 4- (5-cyano-4-{[4- (dimethylcarbamoyl) -2-fluorophenoxy] methyl} -1 H-pyrazol-1-yl) piperidine-1 Carboxylates;

1-methylcyclopropyl 4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine-1 Carboxylates;

4-({5-cyano-1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -1H-pyrazol-4-yl} methoxy) -3-fluoro Rho-N, N-dimethylbenzamide;

1-methylcyclopropyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate ;

tert-butyl (3S, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazol-1-yl)- 3-fluoropiperidine-1-carboxylate;

tert-butyl (3R, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazol-1-yl)- 3-fluoropiperidine-1-carboxylate;

1-methylcyclopropyl (3S, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate;

1-methylcyclopropyl (3R, 4R) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate;

tert-butyl (3S, 4S) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3-fluoro Piperidine-1-carboxylate;

tert-butyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl) -3-fluoro Piperidine-1-carboxylate;

1-methylcyclopropyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3- Fluoropiperidine-1-carboxylate;

1-methylcyclopropyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3- Fluoropiperidine-1-carboxylate;

1-methylcyclopropyl (3R, 4S) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3- Fluoropiperidine-1-carboxylate;

tert-butyl 4- (5-cyano-4-{[4- (1H-1,2,3-triazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperi Dine-1-carboxylate;

tert-butyl 4- (5-cyano-4-{[4- (2H-1,2,3-triazol-2-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperi Dine-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 4- [5-cyano-4-({[1- (methylsulfonyl) piperidin-4-yl] oxy} methyl) -1H-pyrazol-1-yl] piperidine-1 Carboxylates;

tert-butyl 4- [5-cyano-4-({2-fluoro-4-[(2-hydroxyethyl) (methyl) carbamoyl] phenoxy} methyl) -1H-pyrazole-1- General] piperidine-1-carboxylate;

tert-butyl 4- [5-cyano-4-({2-fluoro-4-[(3-hydroxypyrrolidin-1-yl) carbonyl] phenoxy} methyl) -1H-pyrazole- 1-yl] piperidine-1-carboxylate;

tert-butyl 4- (4-{[4- (azetidin-1-ylcarbonyl) -2-fluorophenoxy] methyl} -5-cyano-1H-pyrazol-1-yl) piperidine -1-carboxylate;

1-methylcyclopropyl 4- [5-cyano-4-({[1- (methylsulfonyl) piperidin-4-yl] oxy} methyl) -1H-pyrazol-1-yl] piperidine -1-carboxylate;

1-methylcyclopropyl 4- (5-cyano-4-((2-fluoro-4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -1H-pyrazole -1-yl) piperidine-1-carboxylate;

Isopropyl 4- (5-cyano-4-((2-fluoro-4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -1H-pyrazole-1- I) piperidine-1-carboxylate;

1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-fluoro-4- (1H-1,2,3-triazole-1- Yl) phenoxy) methyl) -1H-pyrazole-5-carbonitrile;

Isopropyl 4- (4-((4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine -1-carboxylate;

4-((4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -1- (1- (5-ethylpyrimidin-2-yl) piperidine-4- Yl) -1H-pyrazole-5-carbonitrile;

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-ethylpyrimidin-2-yl) piperidine-4- Yl) -1H-pyrazole-5-carbonitrile;

1-Methylcyclopropyl 4- (5-cyano-4-((2-fluoro-4- (2H-1,2,3-triazol-2-yl) phenoxy) methyl) -1H-pyrazole -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- I) piperidine-1-carboxylate;

1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-fluoro-4- (2H-1,2,3-triazole-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-pyrazole- 1-yl) piperidine-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) pi Ferridine-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-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((3-fluoro-4- (1H-tetrazol-1-yl) phenoxy) methyl ) -1H-pyrazole-5-carbonitrile;

4-((5- (1H-1,2,3-triazol-1-yl) pyridin-2-yloxy) methyl) -1- (1- (5-ethylpyrimidin-2-yl) piperi Din-4-yl) -1H-pyrazole-5-carbonitrile;

1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-methyl-6- (1H-1,2,3-triazol-1-yl ) Pyridin-3-yloxy) methyl) -1H-pyrazole-5-carbonitrile;

Isopropyl 4- (5-cyano-4-((2-methyl-6- (1H-1,2,3-triazol-1-yl) pyridin-3-yloxy) methyl) -1H-pyrazole -1-yl) piperidine-1-carboxylate;

1-methylcyclopropyl 4- (5-cyano-4-((2-methyl-6- (1H-1,2,3-triazol-1-yl) pyridin-3-yloxy) methyl) -1H -Pyrazol-1-yl) piperidine-1-carboxylate;

1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) Phenoxy) methyl) -1H-pyrazole-5-carbonitrile;

1-Methylcyclopropyl 4- (4-((4- (azetidine-1-carbonyl) -2-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperi Dine-1-carboxylate;

Isopropyl 4- (4-((4- (azetidine-1-carbonyl) -2-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine-1 Carboxylates; And

4-((4- (azetidine-1-carbonyl) -2-fluorophenoxy) methyl) -1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -1H-pyrazole-5-carbonitrile;

Or a pharmaceutically acceptable salt thereof.

These compounds modulate the activity of G-protein-coupled receptors. More specifically, the compound modulates GPR119. As such, the compounds are useful for the treatment of diseases, such as diabetes, in which the activity of GPR119 causes the pathology or symptoms of the disease. Examples of such conditions are hyperlipidemia, type I diabetes, type II diabetes, idiopathic type I diabetes (type Ib), adult latent autoimmune diabetes (LADA), early-onset type 2 diabetes (EOD), adolescents Onset atypical diabetes (YOAD), adolescent onset diabetes (MODY), malnutrition-related diabetes, gestational diabetes, coronary heart disease, ischemic stroke, restenosis after angioplasty, peripheral vascular disease, intermittent claudication, myocardial infarction ( Necrosis and apoptosis), dyslipidemia, postprandial hemostasis, impaired glucose tolerance (IGT) status, fasting plasma glucose disorder status, metabolic acidosis, ketoneosis, arthritis, obesity, osteoporosis, hypertension, congestive heart failure, left ventricular hypertrophy, peripheral arteries Diseases, diabetic retinopathy, macular degeneration, cataracts, diabetic nephropathy, glomerulosclerosis, chronic renal failure, diabetic neuropathy, metabolic syndrome, syndrome X, premenstrual syndrome, coronary heart disease, angina pectoris, Prerequisite, atherosclerosis, transient ischemic attack, stroke, vascular restenosis, hyperglycemia, hyperinsulinemia, hyperlipidemia, hypertriglyceridemia, insulin resistance, impaired glucose metabolism, impaired glucose tolerance, obesity, erectile dysfunction , Skin and connective tissue disorders, foot ulcers and ulcerative colitis, endothelial dysfunction and vascular elastic disorders. The compounds can be used to treat neurological disorders such as Alzheimer's disease, schizophrenia and cognitive disorders. The compounds will also be beneficial for gastrointestinal diseases such as inflammatory bowel disease, ulcerative colitis, Crohn's disease, irritable bowel syndrome and the like. As mentioned above, the compounds can also be used to stimulate weight loss in obese patients, especially patients with diabetes.

Further embodiments of the invention relate to pharmaceutical compositions containing a compound of the invention. Such formulations will typically contain a compound of the invention in admixture with one or more pharmaceutically acceptable excipients. Such formulations may also contain one or more additional pharmaceutical agents. Examples of such agents include anti-obesity agents and / or antidiabetic agents. A further aspect of the invention relates to the use of a compound of the invention in the manufacture of a medicament for the treatment of diabetes and related conditions as described herein.

It is to be understood that both the above summary and the following detailed description are exemplary and explanatory only and are not limitative of the invention as claimed.

DETAILED DESCRIPTION OF THE INVENTION

The invention may be more readily understood by reference to the following detailed description of exemplary embodiments of the invention and the examples included therein.

It is to be understood that the present invention is of course not limited to specific synthetic manufacturing methods which may 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. Plural and singular forms should be treated as interchangeable except where indicated by a number.

a. "Halogen" refers to a chlorine, fluorine, iodine or bromine atom.

b. "C ₁ -C ₄ alkyl" refers to a branched or straight chain alkyl group containing 1 to 5 carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and the like.

c. “C ₁ -C ₄ alkoxy” refers to straight or branched chain alkoxy groups containing 1 to 4 carbon atoms such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy and the like Refer.

d. "C ₃ -C ₆ cycloalkyl" refers to a non-aromatic ring that is fully hydrogenated and exists as a single ring. Examples of such carbocyclic rings include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

e. "C ₁ -C ₄ haloalkyl" refers to a straight or branched chain alkyl group containing 1 to 4 carbon atoms substituted with one or more halogen atoms.

f. "C ₁ -C ₄ haloalkoxy" refers to a straight or branched chain alkoxy group containing 1 to 4 carbon atoms substituted with one or more halogen atoms.

g. "5 to 10 membered heteroaryl" has a total of 5 to 10 ring atoms and contains 1, 2, 3 or 4 heteroatoms independently selected from oxygen, nitrogen and sulfur and wherein 1, 2 or 3 rings (wherein Such rings may be fused). The term "fused" means that the 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 "fused". The term "heteroaryl" refers to aromatic radicals such as pyridine, pyridazine, pyrazine, pyrimidine, imidazo [1,2-a] pyridine, imidazo [1,5-a] pyridine, [1,2,4] tria Solo [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. A "therapeutically effective amount" means (i) treating or preventing a particular disease, condition or disorder, (ii) attenuating, ameliorating or eliminating one or more symptoms of a particular disease, condition or disorder, or (iii) a particular disease described herein. , Amount of a compound of the invention that prevents or delays the onset of one or more symptoms of a condition or disorder.

i. "Patient" refers to warm-blooded animals such as guinea pigs, mice, rats, gerbils, cats, rabbits, dogs, monkeys, chimpanzees and humans.

j. "Treat" includes both prophylactic, ie prophylactic and palliative treatment, ie, alleviating, alleviating or slowing the progression of a patient disease (or condition), or any tissue damage associated with the disease.

k. The term "modulated," "modulating" or "modulate" as used herein, unless otherwise indicated, refers to the activation of G-protein-coupled receptor GPR119 with a compound of the present invention.

l. "Pharmaceutically acceptable" indicates that the substance or composition must be chemically and / or toxicologically compatible with the agent comprising the other ingredients and / or the mammal being treated with it.

m. "Salt" is intended to refer to pharmaceutically acceptable salts and salts suitable for use in industrial processes, such as in the manufacture of compounds.

n. "Pharmaceutically acceptable salt" is intended to refer to either "pharmaceutically acceptable acid addition salt" or "pharmaceutically acceptable basic addition salt" depending on the actual structure of the compound.

o. "Pharmaceutically acceptable acid addition salt" is intended to apply to any nontoxic organic or inorganic acid addition salt of a base compound or any intermediate thereof. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid, and acid metal salts such as sodium orthophosphate, and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include mono-, di- and tricarboxylic acids. Examples of the acid are, for example, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxy -Benzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, 2-phenoxybenzoic acid, p-toluenesulfonic acid, and sulfonic acids such as methane sulfonic acid and 2-hydroxyethane sulfonic acid. Such salts may be present in either hydrate or substantially anhydrous form. In general, acid addition salts of these compounds are soluble in water and various hydrophilic organic solvents.

p. A "pharmaceutically acceptable basic addition salt" is intended to apply to any nontoxic organic or inorganic basic addition salt of a compound or any intermediate thereof. Exemplary bases for forming suitable salts include alkali metal or alkaline-earth metal hydroxides such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide or barium hydroxide; Ammonia, and aliphatic, cycloaliphatic or aromatic organic amines such as methylamine, dimethylamine, trimethylamine and picoline.

q. "Isomer" means "stereoisomers" and "geometric isomers" as defined below. "Stereoisomers" refer to compounds having one or more chiral centers, each of which may exist in an R or S configuration. Stereoisomers include all diastereomeric, enantiomeric and epimeric forms as well as racemates and mixtures thereof. "Geometric isomers" refer to compounds that may exist in cis, trans, anti, syn, engegen (E) and juzammen (Z) forms as well as mixtures thereof.

Certain compounds of the present invention may exist as geometric isomers. The compound may have one or more asymmetric centers and therefore exist in two or more stereoisomeric forms. The present invention includes all individual stereoisomers and geometric isomers of the compounds of the present invention, and mixtures thereof. Individual enantiomers can be obtained by chiral separation or by using the relevant enantiomers in the synthesis. As mentioned above, some of the compounds exist as isomers. These isomeric mixtures can be separated into their individual isomers on the basis of their physicochemical differences by methods known to those skilled in the art, such as by chromatography and / or fractional crystallization. Enantiomers convert enantiomeric mixtures to diastereomeric mixtures by reaction with a suitable optically active compound (e.g., a chiral adjuvant such as chiral alcohol or Mosher acid chloride), to separate diastereomers and , Individual diastereomers can be separated by conversion (eg hydrolysis) to the corresponding pure enantiomers. Enantiomers can also be separated using chiral HPLC columns. Alternatively, certain stereoisomers are synthesized by using optically active starting materials, by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one stereoisomer to another stereoisomer via asymmetric transformation. Can be.

The present invention also encompasses isotopically-labeled compounds of the present invention, which are identical to those shown herein, wherein one or more atoms are replaced by atoms having an atomic mass or mass number that is different from the atomic mass or mass number usually found in nature. Examples of isotopes that may be incorporated in the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, respectively. , ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I, ¹²⁵ I and ³⁶ Cl.

Certain isotopically-labelled compounds of the invention (eg, labeled with ³ H and ¹⁴ C) are useful for compound and / or matrix tissue distribution assays. Certain isotopically labeled ligands, including tritium, ¹⁴ C, ³⁵ S and ¹²⁵ I, may be useful in radioligand binding assays. Tritium (ie ³ H) and carbon-14 (ie ¹⁴ C) isotopes are particularly preferred for ease of preparation and detectability. In addition, substitution with heavier isotopes such as deuterium (ie, ² H) may provide certain therapeutic advantages due to better metabolic stability (eg, increased half-life in vivo or reduced dose requirements), Thus it may be desirable in some situations. Positron emitting isotopes such as ¹⁵ O, ¹³ N, ¹¹ C and ¹⁸ F are useful for positron emission tomography (PET) studies to investigate receptor occupancy. The isotopically labeled compounds of the present invention may be prepared according to procedures similar to those described in the following Schemes and / or Examples herein by replacing isotopically labeled reagents with isotopically labeled reagents.

Certain compounds of the present invention may exist in more than one crystalline form (generally also referred to as “polymorphs”). Polymorphs are, for example, by crystallization under various conditions using different solvents or different solvent mixtures for recrystallization; By crystallization at different temperatures; And / or from very fast cooling to very slow cooling during crystallization. Polymorphs can also be obtained by heating or melting a compound of the invention and then cooling it gradually or rapidly. The presence of the polymorph can be measured by solid probe NMR spectroscopy, IR spectroscopy, differential scanning calorimetry, powder X-ray diffraction or other technique.

In addition, the compounds of the present invention may exist in unsolvated forms 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 as one or more crystalline states, ie cocrystals, polymorphs, or they may exist as amorphous solids. All such forms are included by the present invention and claims.

In embodiments in the compositions of the present invention, the composition further comprises one or more additional pharmaceutical agents selected from the group consisting of anti-obesity agents and anti-diabetic agents. Exemplary anti-obesity agents include dilottapid, mitratapid, implitapide, R56918 (CAS No. 403987), CAS No. 913541-47-6, lorcaserine, cetistastat, PYY _{3 -36} , naltrexone, oleic Oils-estrone, obinetidet, frraminted, tesofensin, leptin, liraglutide, bromocriptine, orlistat, exenatide, AOD-9604 (CAS No. 221231-10-3) and sibutramine . Exemplary antidiabetic agents include metformin, acetohexamide, chlorpropamide, diabines, glybenclamide, glypide, glyburide, glymepiride, glyclazide, glypentide, glyquidone, glysolamide , Tolazamide, tolbutamide, tendamistat, trestatin, acarbose, adifosine, camigliose, emiglitate, miglitol, bogglibose, pramimycin-Q, salvostatin, balaglitazone , Cyglitazone, darglitazone, englitazone, isaglitazone, pioglitazone, rosiglitazone, troglitazone, exendin-3, exendin-4, trodusquemine, reserbatrol, hirthiosal extract, cita Gliptin, bilagliptin, allogliptin and saxagliptin.

In another embodiment of the method of the invention, the compound or composition of the invention is hyperlipidemia, type I diabetes, type II diabetes, idiopathic type I diabetes (type Ib), adult latent autoimmune diabetes (LADA) , Early-onset type 2 diabetes (EOD), adolescent-onset atypical diabetes (YOAD), adolescent onset diabetes (MODY), malnutrition-related diabetes, gestational diabetes, coronary heart disease, ischemic stroke, after angioplasty Restenosis, peripheral vascular disease, intermittent claudication, myocardial infarction (e.g., necrosis and apoptosis), dyslipidemia, postprandial hemostasis, impaired glucose tolerance (IGT) status, fasting plasma glycemic disorder status, metabolic acidosis, ketoneism, arthritis, obesity, Osteoporosis, hypertension, congestive heart failure, left ventricular hypertrophy, peripheral arterial disease, diabetic retinopathy, macular degeneration, cataracts, 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 attack, stroke, vascular restenosis, hyperglycemia, hyperinsulinemia, hyperlipidemia, hypertriglyceridemia, insulin resistance, glucose metabolism Disorders, impaired glucose tolerance, fasting plasma blood glucose disorders, obesity, erectile dysfunction, skin and connective tissue disorders, foot ulcers and ulcerative colitis, endothelial dysfunction and vascular elasticity, hyperapolytic lipoproteinemia, Alzheimer's disease, schizophrenia And cognitive impairment, inflammatory bowel disease, ulcerative colitis, Crohn's disease and irritable bowel syndrome.

In further embodiments, the method further comprises a second composition comprising one or more additional pharmaceutical agents selected from the group consisting of anti-obesity agents and anti-diabetic agents, and one or more pharmaceutically acceptable excipients. Such methods can be used to administer the compositions simultaneously or sequentially and in any order.

In another embodiment, the compounds of the invention are useful in the manufacture of a medicament for treating a disease, condition or disorder that modulates the activity of the G-protein-coupled receptor GPR119. In addition, the compounds are useful in the manufacture of a medicament for the treatment of diabetes or a disease associated with the diabetes.

synthesis

For illustrative purposes, the schemes shown below provide a potential route to synthesize key compounds as well as compounds of the present invention. For a more detailed description of the individual reaction steps, see the Examples section below. One skilled in the art will appreciate that compounds of the present invention may be synthesized using other synthetic routes. Although specific starting materials and reagents are shown in the schemes and discussed below, other starting materials and reagents can be readily substituted to provide various derivatives and / or reaction conditions. In addition, many of the compounds prepared by the methods described below can be further modified in light of the present disclosure using conventional chemistry well known to those skilled in the art.

The compounds of the present invention can be synthesized by synthetic routes, including methods analogous to those well known in the chemical art, in particular in view of the description contained herein. 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 (eg, Louis F. Fieser). and Mary Fieser, Reagents for Organic Synthesis, v. 1-19, Wiley, NY (1967-1999 ed.) or Beilsteins Handbuch der organischen Chemie, 4, Aufl.ed.Springer-Verlag, Berlin (includes appendix) (Also made by the method generally described in the Beilstein online database).

The compounds of the present invention can be prepared using methods similarly known in the art for the preparation of ethers. The reader is interested in textbooks that describe this reaction in more detail, such as:

Headed to.

<Reaction Scheme 1>

Scheme 1 may be used to prepare a compound of Formula N, wherein

이고;X is

ego;

Z is pyrimidine substituted with -C (O) -OR ⁶ , or C ₁ -C ₄ alkyl, CF ₃ , halogen, cyano, C ₃ -C ₆ cycloalkyl or C ₃ -C ₆ cycloalkyl One carbon atom of the cycloalkyl moiety may be optionally substituted with methyl or ethyl;

m is 1, 2 or 3;

n is 0, 1 or 2;

R ¹ is hydrogen, C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl;

R ^2a is hydrogen, fluoro or C ₁ -C ₄ alkyl;

Each R ³ is individually hydroxy, halogen, cyano, C ₁ -C ₄ Alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ Haloalkyl, C ₁ -C ₄ Haloalkoxy, -SO ₂ -R ⁷ , -P (O) (OR ⁸ ) (OR ⁹ ), -C (O) -NR ⁸ R ⁹ , -N (CH ₃ ) -CO-O- (C ₁ -C ₄ ) alkyl, -NH-CO-O- (C ₁ -C ₄ ) alkyl, -NH- Each independently from CO— (C ₁ -C ₄ ) alkyl, —N (CH ₃ ) —CO— (C ₁ -C ₄ ) alkyl, —NH— (CH ₂ ) ₂ —OH, and oxygen, nitrogen, and sulfur Selected from the group consisting of 5 to 6-membered heteroaryl groups containing 1, 2, 3 or 4 heteroatoms selected wherein the carbon atoms on the heteroaryl groups are optionally substituted with R ^4a or nitrogen on the heteroaryl groups The atom is optionally substituted with R ^4b ;

R ^4a is hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ haloalkyl or halogen, wherein said alkyl is optionally substituted with hydroxyl or C ₁ -C ₄ alkoxy;

R ^4b is hydrogen, C ₁ -C ₄ alkyl, —CH ₂ -C ₁ -C ₃ haloalkyl, —C ₂ -C ₄ alkyl-OH or —CH ₂ -C ₁ -C ₄ alkoxy;

R ⁵ is hydrogen, or when R ¹ is hydrogen, R ⁵ is hydrogen or C ₁ -C ₄ alkyl;

R ⁶ is C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl, wherein one carbon atom of the cycloalkyl moiety may be optionally substituted with methyl or ethyl;

R ⁷ represents C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl, NH ₂ or — (CH ₂ ) ₂ —OH;

R ⁸ represents hydrogen or C ₁ -C ₄ alkyl;

R ⁹ is hydrogen, C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl,-(CH ₂ ) ₂ -OH,-(CH ₂ ) ₂ -O-CH ₃ ,-(CH ₂ ) ₃ -OH, -(CH ₂ ) ₃ -O-CH ₃ , 3-oxetanyl or 3-hydroxycyclobutyl;

Or when R ³ is -C (O) -NR ⁸ R ⁹ , R ⁸ and R ⁹ together with the nitrogen atom to which they are attached may form an azetidine, pyrrolidine, piperidine or morpholine ring have.

In step 1, the compound of formula C is formula A in various solvents including but not limited to ethanol, toluene and acetonitrile at temperatures ranging from 22 ° C. to 130 ° C., depending on the solvent used, for a period of 1 to 72 hours. It can be prepared through the condensation reaction of the compound of and commercial compound B (Sigma-Aldrich). If the compound of formula (A) is a hydrogen chloride or trifluoroacetic acid salt, a base modifier such as sodium acetate or sodium bicarbonate may be added in one to three equivalents to neutralize the salt. The reaction can be carried out in polar protic solvents such as methanol and ethanol at temperatures ranging from 22 ° C. to 85 ° C. Typical conditions for this conversion include the use of 3 equivalents of sodium acetate in ethanol heated at 85 ° C. for 3 hours.

Compounds of formula A may be prepared via a four-step procedure starting from substituted or unsubstituted 4-piperidinone hydrochloride salts (J. Med. Chem. 2004, 47, 2180). Firstly these salts are treated with the 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 a reducing agent 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 a compound of formula A, which is typically isolated and corresponding salt (eg, Dihydrochloride salt).

In step 2, the compound of formula D can be prepared via the formation of an intermediate diazonium salt from the compound of formula C via a Sandmeyer reaction (Comp. Org. Synth., 1991, 6, 203). . These salts can be prepared through the diazotization of compounds of formula C using sodium nitrite and aqueous acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and acetic acid alone or in combination. This reaction is typically carried out in water at 0 ° C to 100 ° C. Alternatively, anhydrous conditions using alkyl nitrites such as tert-butyl nitrite with solvents such as acetonitrile can be used at temperatures ranging from 0 ° C. to 95 ° C. (J. Med. Chem. 2006 , 49, 1562]. The diazonium intermediate is then reacted with a copper salt such as copper bromide (II), copper bromide (I), or tribromomethane to form a compound of formula (D). Typical conditions for this conversion include the use of tert-butyl nitrite, copper bromide (II) in acetonitrile at 65 ° C. for 30 minutes.

In step 3, the compound of formula E is a polar aprotic solvent such as tetrahydrofuran, diethyl ether, 1,4-dioxane or 1,2-dime at a temperature ranging from 0 ° C to 110 ° C for 1 to 24 hours. It may be prepared from a compound of formula D using a reducing agent in oxyethane such as lithium aluminum hydride, sodium borohydride, lithium borohydride, borane-dimethylsulfide, borane-tetrahydrofuran. Typical conditions include using borane-dimethylsulfide in tetrahydrofuran at 70 ° C. for 14 hours.

To prepare a compound of formula F from a compound of formula E, a cyano group must be introduced (step 4). This can be accomplished through a range of conditions. One method of introducing cyano groups is a polar aprotic solvent such as N, N-dimethylformamide (DMF), N-methylpyrrolidinone (NMP), at temperatures ranging from 22 ° C. to 200 ° C. for 1 to 24 hours. It may be using copper salts such as copper cyanide in N, N-dimethylacetamide (DMA). Copper cyanide in N, N-dimethylformamide heated at 165 ° C. for 5 hours is a typical protocol for this conversion.

Alternatively in step 4, an alkali sanide salt, such as potassium cyanide or sodium cyanide, can be used to form a polar aprotic solvents such as acetonitrile and dimethylsulfoxide at temperatures ranging from 22 ° C. to 100 ° C. to add cyano groups to the template. Heavy catalysts such as 18-crown-6 (US2005020564) and or tetrabutylammonium bromide (J. Med. Chem. 2003, 46, 1144).

Finally, the use of metal catalysis is common for the conversion shown in step 4. Common cyanide salts used in the catalytic procedure include zinc cyanide, copper cyanide, sodium cyanide and potassium hexacyanoferrate (II). Metal catalysts include copper catalysts such as copper iodide and / or palladium catalysts such as tris (dibenzylideneacetone) dipalladium (Pd ₂ (dba) ₃ ), palladium tetrakis-triphenylphosphine (Pd (PPh ₃ ) ₄ ) or dichloro (diphenyl-phosphinoferrocene) -palladium (Pd (dppf) Cl ₂ ). Such catalysts may be used alone or in any combination with any of the above cyanide salts. Ligands such as 1,1'-bis (diphenylphosphino) -ferrocene (dppf) or metal additives such as zinc or copper metal may be added to this reaction. The reaction is carried out in polar aprotic solvents such as NMP, DMF, DMA with or without water as an additive. The reaction is carried out at temperatures ranging from 22 ° C. to 150 ° C. via conventional heating or microwave heating for 1 to 48 hours, which can be carried out in a sealed or unsealed reaction vessel. Typical conditions for step 4 include using zinc cyanide, Pd ₂ (dba) ₃ , dppf and zinc powder in DMA heated at 120 ° C. under microwaves for 1 hour (J. Med. Chem. 2005 , 48, 1132].

In step 5, the compound of formula G can be synthesized from the compound of formula F via a Mitsunobu reaction. Mitsunobu reactions are described in synthetic literature (see, eg, Chem. Asian. J. 2007, 2, 1340; Eur. J. Org. Chem. 2004, 2763; S. Chem. Eur. J. 2004, 10, 3130]) and a number of synthetic protocols listed in these reviews may be used. Azodicarboxylates such as diethyl azodicarboxylate (DEAD), di-tert-butyl azodicarboxylate (TBAD), diisopropyl azodicarboxylate (DIAD) and phosphine reagents such as tri The use of the Mitsunobu reaction protocol using phenylphosphine (PPh ₃ ), tributylphosphine (PBu ₃ ) and polymer-supported triphenylphosphine (PS-PPh ₃ ) is useful for the compounds of formula F and general structure X-OH. Combined with a compound. Solvents used in the 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 azodicarboxylate used. Can be. Typical conditions for this conversion are the use of DEAD with PS-PPh ₃ in 1,4-dioxane at 22 ° C. for 15 hours.

An alternative to the Mitsunobu reaction for preparing compounds of formula G is to use methanesulfonyl chloride or para-toluenesulfonyl chloride, respectively, in the presence of a base such as triethylamine or pyridine to convert the compound of formula F to the corresponding methanesulfo Conversion to nate or para-toluenesulfonate derivatives. 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 give a compound of formula G.

Compounds of formula K, wherein R ¹ is C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl, can be prepared from the compound of formula F in three steps: 1) the corresponding formula H of the primary alcohol (Step 6, scheme 1), 2) to provide a secondary alcohol of formula J, wherein R ¹ is C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl (step 7 ) Reaction of an aldehyde intermediate of formula H with an organometallic reagent of formula R ¹ M, wherein M is lithium (Li) or magnesium halide (MgCl, MgBr or MgI), and 3) under conditions of Mitsunobu reaction Reaction of the secondary alcohol of with the phenol of formula X-OH (step 8).

In step 6 (Scheme 1), the compound of formula H is 1 to 20 in a solvent alone or in combination including but not limited to dichloromethane, acetonitrile, hexane or acetone at 22 ° C. to 80 ° C. for 1 to 72 hours. It can be formed through an oxidation procedure involving the use of an equivalent amount of activated manganese dioxide. Alternatively, this oxidation is carried out at 0.1 to 1 equivalent of 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) in dichloromethane or chloroform for 0.1 to 12 hours at temperatures ranging from 0 ° C to 22 ° C. Can be carried out using 1 to 3 equivalents of trichloroisocyanuric acid in the presence of). Typical conditions for this conversion are the use of trichloroisocyanuric acid in the presence of 0.1 equivalent of TEMPO in dichloromethane for 1 hour at 22 ° C.

The preparation of compounds of the invention wherein Y is NR ⁵ is also shown in Scheme 1. Compounds of formula L can be prepared from intermediate compounds of formula H (Scheme 1) by reaction with amino compounds of formula X-NH-R ⁵ under reductive amination conditions (step 9) (J. Org. Chem., 1996, 61, 3849; Org.React. 2002, 59, 1]). Similarly, a compound of Formula N, wherein R ¹ is C ₁ -C ₄ alkyl or C ₃ -C ₆ cycloalkyl, is an intermediate of Formula J, wherein R ¹ is C ₁ -C ₄ alkyl or C _3- C ₆ cycloalkyl), 1) oxidation of the formula M to the corresponding ketone (step 10), and 2) reaction of the ketone of formula M with an amino compound of formula X-NH-R ⁵ under reducing amination conditions It can be produced in two steps by (step 11). Alternatively, compounds of Formula L and Formula N, wherein R ⁵ is C ₁ -C ₄ alkyl, are formula (C ₁ -C ₄ ) -Cl, (C ₁ -C ₄ ) -Br in the presence of a base Or from a corresponding compound of formula L, wherein R ⁵ is H, by alkylation with an alkyl halide of (C ₁ -C ₄ ) -I, or a corresponding compound of formula N, wherein R ⁵ is H It can be prepared from).

Compounds of the invention can also be prepared as shown in Schemes 2 and 3, wherein X, Z, R ¹ , R ^2a , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ Is the same as described in Scheme 1. In particular, compounds of formula R may be prepared as shown in Scheme 2.

<Reaction Scheme 2>

In step 1 of Scheme 2, the compound of formula O is dimethyl (diazomethyl) phosphonate or dimethyl- in a solvent including methanol, ethanol or tetrahydrofuran at a temperature ranging from -78 ° C to 22 ° C for 0.1 to 24 hours. It can be formed from the aldehyde of formula H (see also Scheme 1) through the use of 1-diazo-2-oxopropylphosphonate and a base such as potassium carbonate or potassium tert-butoxide. Typical conditions for this conversion include the use of dimethyl-1-diazo-2-oxopropylphosphonate and two equivalents of potassium carbonate in methanol at 22 ° C. for 0.75 hours.

In step 2, the compound of formula Q is formulated via a metal-catalyzed Sonogashira coupling procedure with a compound of general formula XP, wherein P is a halide or trifluoromethsulfonate (triplate) It may be formed from the compound of O. Sonogashira reactions have 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 can be used for the synthesis of compounds of formula Q. Typically, the use of a metal catalyst in the reaction is a copper catalyst such as copper iodide and or a palladium catalyst such as Pd ₂ (dba) ₃ , Pd (PPh ₃ ) ₄ , Pd (dppf) Cl ₂ or Pd (PPh ₃ ) ₂ Cl ₂ . These catalysts may be used alone or in any combination. Base additives are typically used in the reaction and may include amine bases such as diethylamine, triethylamine, diisopropylethylamine or pyrrolidine, or inorganic bases such as potassium carbonate or potassium fluoride. The reaction is carried out in a solvent with or without water as an additive, such as dichloromethane, chloroform, acetonitrile, DMF, toluene or 1,4-dioxane. The reaction is carried out at temperatures ranging from 0 ° C. to 150 ° C. depending on the solvent for a time ranging from 0.1 to 48 hours. Typical conditions for this conversion include the use of CuI and Pd (PPh ₃ ) ₂ Cl ₂ in DMF for 2 hours at 90 ° C.

Finally, in step 3 the compound of formula R can be formed from the compound of formula Q via hydrogenation in the presence of a transition metal catalyst. Common catalysts include the use of 5-20% palladium on carbon, or 5-20% palladium hydroxide on carbon. This reaction is carried out in a Parr shaker device under hydrogen pressure in the range of 1 to 50 psi in a polar solvent such as tetrahydrofuran, ethyl acetate, methanol or ethanol at a temperature of 22 ° C. to 50 ° C. for a time ranging from 0.1 to 24 hours or H. Cube hydrogenation fluidized reactor (ThalesNano, UK). Typical conditions for step 3 include using the compound of formula Q in ethyl acetate at a flow rate of 1 mL / min via a 10% palladium cartridge on carbon on an H-cube fluidized device set to a “full hydrogen” setting.

Scheme 3 shows a process for preparing the compound of formula W.

<Reaction Scheme 3>

In step 1 of Scheme 3, a compound of Formula F (also see Scheme 2) can be treated with a reagent such as phosphorus tribromide or carbon tetrabromide and triphenylphosphine to give a compound of Formula S. The compound of formula S is then reacted in step 2 with triphenylphosphine in a solvent such as dichloromethane, chloroform, toluene, benzene, tetrahydrofuran (THF) or acetonitrile to give the triphenylphosphonium salt of formula T can do. The salts of formula T are then prepared in a solvent such as THF, diethylether or a base in 1,4-dioxane such as n-butyllithium, sodium bis (trimethylsilyl) amide, lithium bis (trimethylsilyl) amide, potassium bis ( It can be combined with a carbonyl compound of formula U in the presence of trimethylsilyl) amide or lithium diisopropylamide to give an alkene compound of formula V, which is typically isolated as a mixture of E and Z geometric isomers (Step 3 ). Such reactions commonly known as Wittig olefination reactions are described in Chem. Rev. 1989, 89, 863; Modern Carbonyl Olefination 2004, 1-17; Liebigs Ann. Chem. 1997, 1283 ]) Is extensively reviewed.

In step 4, the compound of formula W is formed from the compound of formula V via hydrogenation in the presence of a transition metal catalyst. Common catalysts include the use of 5-20% palladium on carbon, or 5-20% palladium hydroxide on carbon. This reaction can be carried out in a similar manner as described in step 3 of Scheme 2.

Alternatively, the compound of formula W may be prepared from the aldehyde of formula H via a Wittich reaction with the triphenylphosphonium salt of formula AA (step 5, scheme 3). As for step 3, this reaction produces an alkene compound of formula V, which is typically isolated as a mixture of E and Z geometric isomers and can be converted to a compound of formula W by hydrogenation. Salts of formula AA are obtained through conversion of the corresponding alcohol to bromide and subsequent reaction with triphenylphosphine in a manner similar to that used to prepare salts of formula T.

The compounds of formula BB shown below, wherein X, Z, R ¹ and R ^2a are as defined in Scheme 1, are secondary alcohols of formula J (see Scheme 2) through a reaction sequence similar to that shown in Scheme 3, or It may be prepared from a ketone of Formula M (see Scheme 2). Conversion of the compound of formula (J) to the corresponding bromide, followed by biti olefination with aldehydes of formula (X-CHO), gives alkenes of formula (CC). Alkenes of the formula CC can also be obtained through the Wittich reaction of a ketone of the formula M with a salt of the general structure X—CH ₂ —PPh ₃ ⁺ Br ⁻ . The alkenes of formula CC are then converted to the compounds of formula BB by hydrogenation.

In certain cases, it is possible to change the order of the steps shown in Schemes 1, 2 and 3. For example, in Scheme 1, it may sometimes be possible to introduce cyano groups onto the pyrazole ring as the final step, ie to reverse the order in which steps 4 and 5 are carried out. In certain cases it is also preferred to introduce or modify the substituents R ³ on the group X, even as a final step in subsequent synthesis. For example, when R ³ is SO ₂ R ⁷ , the SO ₂ R ⁷ group can be formed in the final step by oxidation of the corresponding compound bearing a substituent of the general formula SR ⁷ .

Compounds of the invention are shown in Schemes 1, 2 and 3 starting from 3,3-difluoro-4,4-dihydroxy 1-piperidine carboxylic acid 1,1-dimethylethyl ester (WO 2008121687) It may be prepared according to a similar order as described above. In a manner similar to that described for preparing intermediates of Formula A in Scheme 1, the materials can be converted to hydrazine derivatives of Formula DD, which are then used analogously to intermediates of Formula A in Scheme 1.

As will be readily apparent to those skilled in the art, it may be necessary to protect the distal functional groups of the intermediate (eg, primary or secondary amines). The need for such protection will depend on the nature of the distal functional group and the conditions of the preparation method. Suitable amino-protecting groups (NH-Pg) include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9-fluorenylmethyleneoxycarbonyl (Fmoc) . Similarly, "hydroxy-protecting group" refers to a substituent of a hydroxy group that blocks or protects a hydroxy functional group. Suitable hydroxyl-protecting groups (O-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 general descriptions of protecting groups and their use, see T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

As mentioned above, some of the compounds of the present invention are acidic, which form salts with pharmaceutically acceptable cations. Some of the compounds of the present invention are basic, which forms salts with pharmaceutically acceptable anions. All such salts are within the scope of the present invention, which can be prepared by conventional methods, for example by combining acidic and basic entities in a generally stoichiometric ratio of a suitably aqueous, non-aqueous or partially aqueous medium. . The salt is suitably recovered by filtration, by filtration after precipitation using a non-solvent, by evaporation of the solvent, or by lyophilization in the case of an aqueous solution. The compounds are obtained in crystalline form according to procedures known in the art, for example by dissolution in suitable solvent (s) such as ethanol, hexane or water / ethanol mixtures.

Medical purpose

Compounds of the invention modulate the activity of G-protein-coupled receptor GPR119. As such, the compounds are useful for the prevention and treatment of diseases, such as diabetes, in which the activity of GPR119 causes the pathology or symptoms of the disease. In conclusion, another aspect of the present invention is to administer a compound of the invention, a salt of the compound, or a therapeutically effective amount of a pharmaceutical composition containing such a compound to an individual in need thereof for the treatment of metabolic diseases and / or metabolic-related disorders. And methods of treating metabolic diseases and / or metabolic-related disorders in said individual. Metabolic diseases and metabolic-related disorders include hyperlipidemia, type I diabetes, type II diabetes, idiopathic type I diabetes (type Ib), adult latent autoimmune diabetes (LADA), early-onset type 2 diabetes (EOD) ), Juvenile-onset atypical diabetes (YOAD), juvenile onset diabetes (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, postprandial hemostasis, impaired glucose tolerance (IGT) status, fasting plasma glucose disorder status, metabolic acidosis, ketoneosis, arthritis, obesity, osteoporosis, hypertension, congestive heart failure, left ventricular hypertrophy Peripheral artery disease, diabetic retinopathy, macular degeneration, cataracts, diabetic nephropathy, glomerulosclerosis, chronic renal failure, diabetic neuropathy, metabolic syndrome, syndrome X, premenstrual syndrome, coronary heart quality , Angina, thrombosis, atherosclerosis, myocardial infarction, transient ischemic attack, stroke, vascular restenosis, hyperglycemia, hyperinsulinemia, hyperlipidemia, hypertriglyceridemia, insulin resistance, impaired glucose metabolism, impaired glucose tolerance, fasting plasma glucose status , Obesity, erectile dysfunction, skin and connective tissue disorders, foot ulcers, endothelial dysfunction, hyperlipo B lipoproteinemia and vascular elastic disorders. In addition, the compounds can be used to treat neurological disorders such as Alzheimer's disease, schizophrenia and cognitive disorders. The compounds will also be beneficial for gastrointestinal diseases such as inflammatory bowel disease, ulcerative colitis, Crohn's disease, irritable bowel syndrome and the like. As mentioned above, the compounds can also be used to stimulate weight loss in obese patients, especially patients with diabetes.

In accordance with the above, the present invention further comprises administering a therapeutically effective amount of a compound of the invention to a subject in need of preventing or alleviating the symptoms of any of the diseases or disorders described above. A method of preventing or alleviating the symptoms of a disease or disorder is provided. A further aspect of the invention includes the manufacture of a medicament for the treatment of diabetes and its associated co-morbidity.

In order to exhibit the therapeutic properties described above, the compound needs to be administered in an amount sufficient to modulate the activation of the G-protein-coupled receptor GPR119. Such amounts may vary depending on the particular disease / condition to be 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 inherent disease states of the patient. When administered systemically, the compound typically exhibits its effect in a dosage range of about 0.1 mg / kg / day to about 100 mg / kg / day for any of the diseases or conditions listed above. Repeated daily administration may be desirable, which will vary depending on the condition outlined above.

The compounds of the present invention can be administered by a variety of routes. The compound may be administered orally. The compounds may also be administered parenterally (ie, subcutaneously, intravenously, intramuscularly, intraperitoneally or intradural), rectally or topically.

Administration

The compounds of the present invention may also be used with other pharmaceutical agents for the treatment of the diseases, conditions and / or disorders described herein. Thus, there is also provided a method of treatment comprising administering a compound of the invention in combination with another pharmaceutical agent. Suitable pharmaceutical agents that can be used in combination with the compounds of the present invention include anti-obesity agents (including appetite suppressants), antidiabetic agents, antihyperglycemic agents, lipid lowering agents and antihypertensive agents.

Suitable antidiabetic agents include acetyl-CoA carboxylase-2 (ACC-2) inhibitors, diacylglycerol O-acyltransferase 1 (DGAT-1) inhibitors, phosphodiesterase (PDE) -10 inhibitors, sulfonyl Urea (e.g. acetohexamide, chlorpropamide, diabines, glybenclamide, glyphide, glyburide, glymepiride, glyclazide, glypentide, glyquidone, glysolamide, tall Lazamide and tolbutamide), meglitinide, α-amylase inhibitors (eg, tenamistat, trestatin and AL-3688), α-glucoside hydrolase inhibitors (eg, acarbose) , α-glucosidase inhibitors (e.g. adiposin, camigliose, emiglytate, miglitol, bogliose, pradimycin-Q and salvostatin), PPARγ agonists (e.g., balaglita John, Ciglitazone, Darglitazone, Englitazone, Isaglitazone, Piogli Zone, rosiglitazone and troglitazone), PPAR α / γ agonists (eg CLX-0940, GW-1536, GW-1929, GW-2433, KRP-297, L-796449, LR-90, MK-0767 and SB-219994), biguanides (eg metformin), glucagon-like peptide 1 (GLP-1) agonists (eg exendin-3 and exendin-4), protein tyrosine phosphatase-1B ( PTP-1B) inhibitors (eg, trodusquemin, hirthiosal extract, and Zhang, S., et al., Drug Discovery Today, 12 (9/10), 373-381 (2007)) Compounds described), SIRT-1 inhibitors (eg, reservatrol), dipeptidyl peptidase IV (DPP-IV) inhibitors (eg, citagliptin, bilagliptin, allogliptin and saxaggle Liptin), insulin secretagogues, fatty acid oxidation inhibitors, A2 antagonists, c-jun amino-terminal kinase (JNK) inhibitors, insulin, insulin mimetics, glycogen phosphorylase inhibitors, VPAC2 receptor agonists and SGLT2 inhibitors (sodium dependent And a base course transporter inhibitors, such as the such as rosin pageul ripple). Preferred antidiabetic agents are metformin and DPP-IV inhibitors (eg, citagliptin, bilagliptin, allogliptin and saxagliptin).

Suitable anti-obesity agents are 11β-hydroxy steroid dehydrogenase-1 (11β-HSD type 1) inhibitors, stearoyl-CoA desaturase-1 (SCD-1) inhibitors, MCR-4 agonists, cholecystokinin- A (CCK-A) agonists, monoamine reuptake inhibitors (such as sibutramine), sympathomimetic, β ₃ adrenergic agonists, dopamine agonists (such as bromocriptine), melanocyte-stimulating hormone analogs, 5HT2c efficacy Agents, melanin enrichment hormone antagonists, leptin (OB protein), leptin analogs, leptin agonists, galanin antagonists, lipase inhibitors (such as tetrahydrolipstatin, ie orlistat), appetite suppressants (such as bombesin agonists), neuros -Y peptide antagonists (e.g., NPY Y5 antagonists), PYY _{3 -36} (including analogs thereof), thyroid hormone mimetics, epi dehydroacetic Andros testosterone or an analogue, glucocorticoid agonists or antagonists, orexin antagonists, articles Luca Similar peptide-1 agonists, ciliary neurotrophic factors (such as Regeneron Pharmaceuticals, Inc., Tarrytown, NY) and Procter & Gamble Company (Cincinnati, Ohio). Axokine ™, human aguti-related protein (AGRP) inhibitors, ghrelin antagonists, histamine 3 antagonists or inverse agonists, neuromedin U agonists, MTP / ApoB inhibitors (e.g. Selective MTP inhibitors such as dilottapid), opioid antagonists, orexin antagonists and the like.

Preferred anti-obesity agents for use in the combination aspects of the present invention are gut-selective MTP inhibitors (e.g., dilottapid, mitratapid and implitapid, R56918 (CAS No. 403987) and CAS No. 913541- 47-6), CCKa agonists (eg, N-benzyl-2- [4- (1H-indol-3-ylmethyl) -5-oxo-1-phenyl-4,5-dihydro-2, 3,6,10b-tetraaza-benzo [e] azulen-6-yl] -N-isopropyl-acetamide (described in PCT Publication No. WO 2005/116034 or US Publication No. 2005-0267100 A1), 5HT2c (e. g., serine Mallorca) agonists, MCR4 agonist (e.g., compounds described in US 6,818,658), lipase inhibitor (e.g., a three-start Tilly), PYY _{3 -36} (used herein Likewise, “PYY _{3 -36} ” refers to analogs such as PEGylated PYY _{3 -36} , including those described in US Publication 2006/0178501, opioid antagonists (eg naltrexone), oleoyl-estrone ( CAS number 180003-17-2), Obinetidet (TM30338), Pramlinet (Symlin ^® ), Tesofensin (NS2330), Leptin, Liraglutide, Bromocriptine, Orlistat, Exenatide (Byetta ^® ) , AOD-9604 (CAS No. 221231-10-3) and sibutramine Preferably the compounds and combination therapies of the invention are administered in combination with exercise and a rational diet.

All US patents and publications cited above are incorporated herein by reference.

Pharmaceutical preparation

The invention also provides pharmaceutical compositions comprising a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof in admixture with one or more pharmaceutically acceptable excipients. The compositions include those in forms adapted for oral, topical or parenteral use, which can be used for the treatment of diabetes and related conditions as described above.

The composition may be formulated for administration by any route known in the art, such as subcutaneous, inhalation, oral, topical, parenteral and the like. The composition 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 forms, which include conventional excipients such as binders such as syrup, acacia, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone; Fillers such as lactose, sugar, corn starch, calcium phosphate, sorbitol or glycine; Tableting lubricants such as magnesium stearate, talc, polyethylene glycol or silica; Disintegrants such as potato starch; Or acceptable wetting agents, such as sodium lauryl sulfate. Tablets may be coated according to methods well known in conventional pharmaceutical practice.

Oral liquid formulations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as dry products which are reconstituted with water or other suitable vehicle before use. Such liquid preparations are conventional additives such as suspending agents, for example sorbitol, methyl cellulose, glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel or hydrogenated edible fats, emulsifiers such as lecithin, sorbent Non-elastic 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 such as 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 using compounds and sterile vehicles, with water being preferred. Depending on the vehicle and concentration used, the compounds may be suspended or dissolved in the vehicle or other suitable solvent. In preparing the solution, the compound is dissolved in water for injection and sterile filtered, then filled into a suitable vial or ampoule and sealed. Advantageously, agents such as local anesthetics, preservatives, buffers and the like can be dissolved in the vehicle. To improve the stability, the composition can be filled into vials and then frozen, and the water removed under vacuum. The dry lyophilized powder may then be sealed in a vial and supplied with a vial of water for injection to reconstitute the liquid prior to use. Parenteral suspensions are prepared in substantially the same manner except that the compounds are suspended instead of dissolved in the vehicle and sterilization cannot be performed by filtration. The compound can be sterilized by exposure to ethylene oxide and then suspended in a sterile vehicle. Advantageously, a surfactant or wetting agent is included in the composition to facilitate even distribution of the compound.

The composition may contain, for example, from about 0.1% to about 99% by weight of active substance, depending on the method of administration. If the composition comprises dosage units, each unit will contain, for example, about 0.1 to 900 mg of active ingredient, more typically 1 mg to 250 mg of active ingredient.

The compounds of the present invention can be formulated for administration in any convenient manner for use in human or veterinary medicine, similar to other antidiabetic agents. Such methods are known in the art and are summarized above. For a more detailed discussion relating to the preparation of such formulations, reader's attention is directed to Remington's Pharmaceutical Sciences, 21 ^st Edition, by University of the Sciences in Philadelphia.

Embodiments of the invention are illustrated by the following examples. However, it should be understood that embodiments of the present invention are not limited to the specific details of these examples, as other variations of embodiments of the present invention are known to those skilled in the art or will be apparent in light of the present disclosure.

Example

Unless otherwise specified, starting materials are generally commercial sources, such as Aldrich Chemicals Co. (Milwaukee, WI), Lancaster Synthesis, Inc. (Lancaster Synthesis, Inc .; Wyndham, New Hampshire), Acros Organics (Fairlon, NJ), Maybridge Chemical Company, Limited (Maybridge Chemical Company, Ltd .; Cornwall, UK), Tiger Scientific ( Tyger Scientific, Princeton, NJ and AstraZeneca Pharmaceuticals (London, UK), Mallinckrodt Baker (Phillipsburg, NJ); Available from EMD (Gibbstown, NJ).

General Experiment Procedure

NMR spectra were either Varian Unity ™ 400 (DG400-5 probe) or 500 (DG500-5 probe) at 400 MHz or 500 MHz, respectively, at room temperature for proton analysis (both Varian Inc. (Available from Palo Alto, CA). Chemical shifts are expressed in parts per million (delta) relative to residual solvent as internal reference. Peak shape is indicated as follows: s, singlet; d, doublet; dd, doublet of doublets; t, triplet; q, quadrature; m, multiplet; bs, broad singlet; 2s, two singlets.

Atmospheric pressure chemical ionization mass spectra (APCI) were measured using a 50:50 water / acetonitrile eluent system at a flow rate of 0.3 mL / min (Waters ™ spectrometer (Micromass ZMD, carrier gas: nitrogen) ( Obtained on Waters Corp. (available from Milford, Mass.) Electrospray ionization mass spectra (ES) were obtained with a 95: 5-0: 100 gradient of acetonitrile, with 0.01% formic acid added to each solvent. Heavy water was obtained on a liquid chromatography mass spectrometer from a Waters ™ (Micromass ZQ or ZMD instrument (carrier gas: nitrogen) (Waters Corporation, Milford, Mass.). The instrument was subjected to 1 mL / min for 3.75 minutes, Or a Varian Polaris 5 C18-A20x2.0 mm column (Varian Inc., Palo, CA) at a flow rate of 2 mL / min for 1.95 minutes. Alto) was used.

Column chromatography was performed on silica gel under nitrogen pressure using a Flash 40 Biotage ™ column (ISC, Inc., Shelton, Connecticut) or Biotage ™ SNAP cartridge KPsil or Redisep ) Rf silica (obtained from Teledyne Isco Inc.) Preparative HPLC was performed using a Waters FractionLynx system with a photodiode array (Waters 2996) and a mass spectrometer ( Analytical HPLC operations were performed using a Waters 2795 Alliance HPLC or Waters ACQUITY UPLC (Photodiode Array, Single Quadrupole Mass and Vapor). Light scattering detection was performed.

Concentration under vacuum refers to evaporation of the solvent under reduced pressure using a rotary evaporator.

Unless otherwise indicated, chemical reactions were carried out at room temperature (about 23 ° C.). In addition, unless otherwise indicated, the chemical reaction proceeded under a nitrogen atmosphere.

Pharmacological data

The practice of the invention for the treatment of diseases modulated by agonist activation of G-protein-coupled receptor GPR119 using the compounds of the invention is demonstrated by activity in one or more of the functional assays described herein below. Can be. Sources are provided in parentheses.

In vitro functional assay

β-lactamase:

Assays for GPR119 agonists use cell-based (hGPR119 HEK293-CRE beta-lactamase) reporter constructs, wherein agonist activation of human GPR119 is dependent upon beta-lactamase production and cyclic AMP response elements (CRE). Connected. GPR119 activity was then measured using a FRET-enabled beta-lactamase substrate, CCF4-AM (Live Blazer FRET-B / G loading kit (Invitrogen Cat. No. K1027)). Specifically, hGPR119-HEK-CRE-beta-lactamase cells (Invitrogen 2.5 × 10 ⁷ cells / mL) were harvested from the liquid nitrogen reservoir and plated medium (Dulbecco's modified Eagle's medium high glucose (DMEM: Gibco ( Gibco) Catalog No. 11995-065), 10% Heat Inactivated Fetal Bovine Serum (HIFBS; Sigma Catalog No. F4135), 1X MEM Non-Essential Amino Acids (Gibco Catalog No. 15630-080), 25 mM HEPES (pH 7.0) (Gibco Catalog No. 15630-080), 200 nM potassium clavulanate (Sigma catalog number P3494). Cell concentration was adjusted using cell plating medium, and 50 μL (12.5 × 10 ⁴ viable cells) of the cell suspension was placed in a black transparent bottomed poly-d-lysine coated 384-well plate (Grainer Bio-One) Greiner Bio-One) Cat. No. 781946) and incubated at 37 ° C. in a humid environment containing 5% carbon dioxide. After 4 hours, the plating medium was removed and replaced with 40 μL of assay medium (the assay medium is a plating medium without potassium clavulanate and HIFBS). The concentration of each compound to be tested was then added in varying concentrations of 10 μL (final DMSO ≦ 0.5%) and the cells were incubated for 16 hours at 37 ° C. in a humid environment containing 5% carbon dioxide. The plate was removed from the incubator and allowed to equilibrate to room temperature for approximately 15 minutes. 10 μL of 6 × CCF4 / AM working dye solution (prepared according to instructions in Live Blazer FRET-B / G Loading Kit (Invitrogen Catalog No. K1027)) was added to each well and incubated in the dark for 2 hours at room temperature. . Fluorescence was measured on an EnVision fluorometric plate reader (excitation 405 nm, emission 460 nm / 535 nm). EC ₅₀ was determined from agonist-response curves analyzed with a curve fitting program using a four-parameter logistic dose-response equation.

cAMP:

GPR119 agonist activity was also determined in a cell-based assay using HTRF (homogeneous time-resolved fluorescence) cAMP detection kit (cAMP Dynamic 2 assay kit; Cis Bio catalog number 62AM4PEC) to measure cAMP levels in cells. . The method is a competitive immunoassay between native cAMP produced by the cell and cAMP labeled with the dye d2. Tracer binding is visualized by Mab anti-cAMP labeled with Cryptate. Specific signals (ie energy transfer) are inversely proportional to cAMP concentration in a standard or sample.

Specifically, hGPR119 HEK-CRE beta-lactamase cells (invitrogen 2.5 × 10 ⁷ cells / mL; same cell line as used in the beta-lactamase assay described above) were harvested from cryopreservation and growth medium (Dulbecco) Modified Eagle Medium High Glucose (DMEM: Gibco Catalog No. 11995-065), 1% Charcoal Dextran Treated Fetal Bovine Serum (CD Serum; HyClone Catalog No. SH30068.03), 1x MEM Non-Essential Amino Acids (Gibco Catalog No. 15630-080) and 25 mM HEPES (pH 7.0) (Gibco Cat. No. 15630-080). Cell concentration was adjusted to 1.5 × 10 ⁵ cells / mL and 30 mL of the suspension was added to a T-175 flask and incubated at 37 ° C. in a humid environment in 5% carbon dioxide. After 16 hours (overnight), the cells (tap on the flask side) were removed from the T-175 flask and centrifuged at 800 xg, followed by assay medium (1x HBSS + CaCl ₂ + MgCl ₂ (Gibco Catalog No. 14025-092). ) And 25 mM HEPES (pH 7.0) (Gibco Cat. No. 15630-080). Adjust the cell concentration to 6.25 x 10 ⁵ cells / mL using the assay medium, and add 8 μL (5000 cells) of the cell suspension to a white grinder 384-well, low volume assay plate (VWR Catalog No. 82051-458). To each well.

The concentration of each compound to be tested was varied and diluted in 3-isobutyl-1-methylxanthine containing assay buffer (IBMX; Sigma catalog number I5879) and added to the assay plate wells in 2 μL volume (final IBMX concentration). Is 400 μΜ and the final DMSO concentration is 0.58%). After 30 min incubation at room temperature, 5 μL of labeled d2 cAMP and 5 μL of anti-cAMP antibody (both diluted 1:20 in cell lysis buffer; as described in the manufacturer's assay protocol) were determined for each assay plate. Was added to the wells. 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 330 nm and emission 615 and 665 nm. Agonist-response curves converted from raw cAMP data to nM cAMP by interpolation from the cAMP standard curve (as described in the manufacturer's assay protocol) and analyzed with a curve fitting program using 4-parameter logistic dose-response equations. EC 50 was determined from.

It is recognized that the cAMP response due to the activation of GPR119 can be generated in cells other than the specific cell lines used herein.

β-arrestin:

GPR119 agonist activity is also measured in cell-based assays using the DiscoverX PathHunter β-arrestin cell assay technology and its U2OS hGPR119 β-arrestin cell line (DiscoverX catalog number 93-0356C3). It was. In this assay, agonist activation was determined by measuring the interaction of agonist-induced β-arrestin with activated GPR119. A small enzyme fragment of 42 amino acids, called ProLink, was added to the C-terminus of GPR119. Arestin was fused to a larger enzyme fragment named EA (enzyme receptor). Activation of GPR119 stimulated the binding of Arestin and made the two enzyme fragments complementary, resulting in the formation of a functional β-galactosidase enzyme capable of hydrolyzing the substrate and generating chemiluminescent signals.

Specifically, U2OS hGPR119 β-arrestin cells (DiscoverX 1 x 10 ⁷ pcs / mL) were harvested from cryopreservation, growth medium (minimum essential medium (MEM; Gibco Cat. No. 11095-080), 10% heat Activated fetal bovine serum (HIFBS; Sigma catalog number F4135-100), 100 mM sodium pyruvate (Sigma catalog number S8636), 500 μg / mL G418 (Sigma catalog number G8168), and 250 μg / mL Hygromycin B ( Invitrogen catalog number 10687-010) Adjust the cell concentration to 1.66 × 10 ⁵ cells / mL and add 30 mL of the suspension to a T-175 flask and 37 ° C. in a humid environment in 5% carbon dioxide. After 48 hours, cells were removed from the T-175 flask with enzyme-free cell dissociation buffer (Gibco Cat. No. 13151-014), centrifuged at 800 × g, and then plated medium (Opti). -MEM I (Invitrogen / BRL Catalog No. 31985-070) and 2% charcoal dextran treated fetal bovine serum (CD serum; Hyclone catalog number SH30068.03) Cell concentrations were 2.5 × 10 ⁵ cells / mL using plating medium. 10 μL (2500 cells) of the cell suspension is added to each well of a white grinder 384-well, low volume assay plate (VWR Cat. No. 82051-458), and the plate is placed in a humid environment in 5% carbon dioxide. Incubate at 37 ° C.

After 16 hours (overnight), the assay plate was removed from the incubator and assayed for (assay buffer (1 × HBSS + CaCl ₂ + MgCl ₂ (Gibco catalog number 14025-092), 20 mM HEPES pH 7.0) (Gibco catalog number 15630-080). Concentrations of each compound to be tested (diluted in 0.1% BSA (Sigma Catalog No. A9576)) were added to the assay plate wells in varying volumes of 2.5 μL (final DMSO concentration was 0.5%) in 5% carbon dioxide. After 90 minutes incubation at 37 ° C. in a humid environment, the Galacton Star β-galactosidase substrate (PathHunter Detection Kit (DiscoverX Catalog No. 93-0001)) (manufacturer of 7.5 μL was added to each well of the assay plate The plate was incubated at room temperature and after 60 minutes the change in luminescence was Envision 2104 multilabel at 0.1 seconds per well. Read using a plate reader EC50 was determined from 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 Assays

Wild-type human GPR119 (published in PCT Patent Publication No. 2010/106457) was used as pIRES-Puro-hGPR119 as a template and the following primers for polymerase chain reaction (PCR) (Pfu Turbo Masterer Mix, Stratazine ( Amplified by Stratagene (La Jolla, CA):

hGPR119 BamH1, top

 (BamH1 site insert at the 5 'end)

hGPR119 EcoRI, sub

(EcoRI site insert at 3 'end)

Purified amplified product (Qiaquick Kit (Qiagen, Valencia, CA)), BamH1 and EcoRI (New England BioLabs; Ipswich, Mass.) According to manufacturer's protocol The vector pFB-VSVG-CMV-poly (published in PCT Patent Publication No. 2010/106457) was digested with BamH1 and EcoRI (New England Biolabs, Ipswich, Mass.) Digested DNA was treated with 1% agarose. Separation by electrophoresis on the gel; fragments were cut from the gel and purified (Qiaquik Kit (Qiagen, Valencia, CA)), ligation of the vector and gene fragments (Rapid Ligase Kit) (Roche, Pleasanton, CA), were transformed into one-shot DH5alpha T1R cells (Invitrogen, Carlsbad, CA.) Eight ampicillin-resistant colonies (“clones 1-8”) were miniffed. (QIAGEN mini prep kit (Qiagen Miniprep Kit) (Qiagen; Valencia, Calif.)) Was grown for, and sequence analysis confirmed his identity and the correct insertion orientation.

PFB-VSVG-CMV-poly-hGPR119 construct (clone # 1) was transformed into one shot DH10Bac cells (Invitrogen, Carlsbad, CA) according to the manufacturer's protocol. Eight positive (ie white) colonies were again streaked to identify "positive" and subsequently grown for bakmid isolation. Recombinant hGPR119 bakmid was isolated via a modified alkaline lysis procedure using buffers from the Qiagen Miniprep Kit (Qiagen, Valencia, CA). Briefly, pelleted cells were lysed in Buffer P1, neutralized in Buffer P2, and precipitated with Buffer N3. The precipitate was pelleted by centrifugation (17,900 × g for 10 minutes) and the supernatant was combined with isopropanol to precipitate DNA. DNA was pelleted by centrifugation (17,900 × g for 30 minutes), washed once with 70% ethanol and resuspended in 50 μL of buffer EB (Tris-HCL, pH 8.5). Polymerase chain reaction (PCR) using commercially available primers (M13F, M13R, Invitrogen, Carlsbad, CA) was used to confirm the presence of hGPR119 insert in bakmid.

hGPR119 recombinant baculovirus generation

Generation of P0 Virus Stock

Suspension-adapted Sf9 cells grown in Sf900II medium (Invitrogen, Carlsbad, CA) were transfected with 10 μL of hGPR119 bakmid DNA according to the manufacturer's protocol (Cellfectin, Invitrogen, Carlsbad, CA). Infected. After 5 days incubation, the conditioned medium (ie, "P0" virus stock) was centrifuged and filtered through a 0.22 μm filter (Steriflip, Millipore, Billerica, Mass.).

Generation of frozen virus (BIIC) stock solutions

For long-term virus storage and production of working (ie, "P1") virus stocks, frozen BIIC (baculovirus infected insect cells) stocks were generated as follows: Suspension-fitted Sf9 cells were prepared in Sf900II medium (Invitro Trogen, Carlsbad, CA) and were infected with stocks of hGPR119 P0 virus. After 24 hours growth, infected cells were slowly centrifuged (approximately 100 xg) and resuspended in freezing medium (10% DMSO in Sf900II medium, 1% albumin) to a final density of 1 x 10 ⁷ cells / mL, Frozen in 1 mL aliquots according to standard freezing protocol.

Generation of Working ("P1") Viral Stock

Suspension adapted Sf9 cells grown in Sf900II medium (Invitrogen, Carlsbad, Calif.) Were infected with a 1: 100 dilution of thawed hGPR119 BIIC stock solution and incubated for several days (shaking at 27 ° C.). When the viability of the cells reached 70%, the conditioned media was collected by centrifugation and virus titers determined by ELISA (BaculoElisa Kit, Clontech, Mountain View, CA). It was.

Overexpression of hGPR119 in suspension-adapted HEK 293FT cells

HEK 293FT cells (Invitrogen, Carlsbad, CA) were grown in 293 Freestyle medium (Invitrogen) supplemented with 50 μg / mL neomycin and 10 mM HEPES in shake flasks (37 ° C., 8 % Carbon dioxide, shaking). Centrifuge the cells slowly (approximately 500 × g, 10 min) and resuspend the pellet in a mixture of Dulbecco's PBS (minus Mg ++ /-Ca ++) supplemented with 18% fetal bovine serum (Sigma Aldrich) and P1 virus to infect multiple The degree (MOI) was 10 and the final cell density was 1.3 × 10 ⁶ cells / mL (total volume 2.5 L). Cells were transferred to a 5 L Wave Bioreactor Wavebag (Wave Technologies, Massachusetts) and incubated at 27 ° C. for 4 hours (17 vibrations / min, 7 degree platform angle); At the end of the incubation period, an equal amount (2.5 L) of 293 freestyle medium supplemented with 30 mM sodium butyrate (Sigma Aldrich) was added (final concentration = 15 mM) and cells were grown for 20 hours (37 ° C., 8%). CO ₂ [0.2 L / min], 25 vibrations / min, 7 degree platform angle). Cells were harvested by centrifugation (3,000 × g, 10 min), washed once on DPBS (minus Ca ++ / Mg ++), resuspended in 0.25 M sucrose, 25 mM HEPES, 0.5 mM EDTA, pH 7.4, Frozen at -80 ° C.

Membrane Preparation for Radioligand Binding Assay

Frozen cells were thawed on ice and centrifuged at 700 xg (1400 rpm) for 10 minutes at 4 ° C. Cell pellets were resuspended in 20 mL phosphate-buffered saline and centrifuged at 1400 rpm for 10 minutes. The cell pellet was then subjected to homogenization buffer (10 mM HEPES (Gibco # 15630), pH 7.5, 1 mM EDTA (BioSolutions, # BIO260-15), 1 mM EGTA (Sigma, # E-4378), 0.01 mg / mL Benzamidine (Sigma #B 6506), 0.01 mg / mL Bacitracin (Sigma #B 0125), 0.005 mg / mL Lupeptin (Sigma #L 8511), 0.005 mg / mL Aprotinin (Sigma #A 1153 Resuspend in)) and incubate on ice for 10 minutes. The cells were then lysed in 15 smooth strokes with a fitted glass Dounce homogenizer. Homogenates were centrifuged at 1000 xg (2200 rpm) for 10 minutes at 4 ° C. The supernatant was transferred to a new centrifuge tube on ice. Cell pellets were resuspended in homogenization buffer, centrifuged again at 1000 × g (2200 rpm) for 10 minutes at 4 ° C., then the supernatant was removed and the pellet was resuspended in homogenization buffer. Repeat the above procedure three times, then combine the supernatants and add Benzonase (Novagen # 71206) and MgCl ₂ (Fluka # 63020) to final concentrations of 1 U / mL and 6, respectively. Add in mM and incubate on ice for 1 hour. The solution was then centrifuged at 25,000 × g (15000 rpm) for 20 minutes at 4 ° C., the supernatant was discarded, and the pellet was resuspended in fresh homogenization buffer (minus benzonase and MgCl ₂ ). After repeating the 25,000 xg centrifugation step, the final membrane pellet was resuspended in homogenization buffer and frozen at -80 ° C. Protein concentration was determined using the Pierce BCA Protein Assay Kit (Pierce Reagent A # 23223 and B # 23224).

Synthesis and Purification of [ ³ H] -Compound A

Compound A (Isopropyl 4- (1- (4- (methylsulfonyl) phenyl) -3a, 7a-dihydro-1H-pyrazolo [3,4-d] pyrimidin-4-yloxy) piperidine -1-carboxylate, as shown above) (4 mg, 0.009 mmol) is dissolved in 0.5 mL of dichloromethane and the resulting solution is (1,5-cyclooctadiene) (pyridine) (tricyclohexylphosphate) Pin) -iridium (I) hexaflurophosphate (J. Organometal. Chem. 1979, 168, 183) (5 mg, 0.006 mmol). The reaction vessel was sealed and the solution stirred for 17 hours under tritium gas atmosphere. The reaction solvent was removed under reduced pressure and the resulting residue was dissolved in ethanol. Purification of crude [ ³ H] -Compound A was carried out by preparative HPLC using the following conditions.

Column: Atlantis, 4.6 x 150 mm, 5 μm

Mobile phase A: water / acetonitrile / formic acid (98/2 / 0.1)

Mobile Phase B: Acetonitrile

Gradient: time% B

0.00 30.0

1.00 30.0

13.00 80.0

Run time: 16 minutes

After time: 5 minutes

Flow rate: 1.5 mL / min

Injection volume: 20-50 μL

Injection Solvent: DMSO

Detection: UV at 210 nm and 245 nm

The specific activity of the purified [ ³ H] -Compound A was determined to be 70 Ci / mmol by mass spectrometry.

Alternatively, binding assays can be performed using [ ³ H] -Compound B.

Synthesis and Purification of [ ³ H] -Compound B

Compound B (tert-butyl 4- (1- (4- (methylsulfonyl) phenyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yloxy) piperidine-1-carboxylate (As indicated above) (5 mg, 10.6 μmol) was dissolved in 1.0 mL of dichloromethane and the resulting solution was treated with crabtree catalyst (5 mg, 6.2 μmol). The reaction vessel was sealed and the solution stirred for 17 hours under tritium gas atmosphere. The reaction solvent was removed under reduced pressure and the resulting residue was dissolved in ethanol. Purification of crude [ ³ H] -Compound B 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. Was performed.

The specific activity of the purified [ ³ H] -Compound B was determined to be 57.8 Ci / mmol by mass spectrometry.

GPR119 Radioligand Binding Assay

Test compounds were serially diluted in 100% DMSO (J.T. Baker # 922401). 2 μL of each dilution was added to the appropriate wells of a 96-well plate (each concentration measured in triplicate). Unlabeled Compound A (or Compound B) at a final concentration of 10 μM was used to determine non-specific binding.

[ ³ H] -Compound A (or [ ³ H] -Compound B) was combined with a buffer (50 mM Tris-HCl, pH 7.5), (Sigma # T7443), 10 mM MgCl ₂ (Fluka 63020), 1 mM EDTA (Biosolutions # BIO260-15), 0.15% bovine serum albumin (Sigma # A7511), 0.01 mg / mL benzamidine (Sigma #B 6506), 0.01 mg / mL bacitracin (Sigma #B 0125), 0.005 mg Dilute to a concentration of 60 nM in / mL leupetin (Sigma #L 8511), 0.005 mg / mL aprotinin (Sigma #A 1153), and 100 μL in a 96-well plate (Nalge Nunc # 267245) To all wells.

Membranes expressing GPR119 were thawed, diluted in binding buffer to a final concentration of 20 μg / 100 μL per well, and 100 μL of diluted membrane was added to each well of a 96-well plate.

The plate was incubated for 60 minutes with shaking at room temperature (approximately 25 ° C.). The assay was terminated by vacuum filtration on pre-soaked GF / C filter plates (Packard # 6005174) in 0.3% polyethyleneamine using a Packard harvester. The filter was then washed six times using wash buffer (50 mM Tris-HCl, pH 7.5 maintained at 4 ° C). The filter plate was then air-stained overnight at room temperature. Add 30 μL of scintillation liquid (Ready Safe (Beckman Coulter # 141349) to each well, seal the plate, and use a Wallac Trilux MicroBeta plate-based scintillation counter The radioactivity associated with each filter was measured.

By carrying out the saturation binding ^[3 H] - Compound A were analyzed data by a linear regression (Prism Graph Pad (- determining the Kd for (or ^[3 H] compound B), in an appropriate ratio to the 1-site hyperbolic Graph Pad Prism). IC ₅₀ was determined from competition curves analyzed with the patented curve fitting program (SIGHTS) and the four-parameter logistic dose response equation. Ki values were calculated from IC ₅₀ values using the Cheng-Prusoff equation.

The following results were obtained for beta-lactamase, beta-arrestin, cAMP and binding assays:

^* Intrinsic activity is standard GPR119 agonist at final concentration of 10 μM, 4-[[6-[(2-fluoro-4 methylsulfonylphenyl) amino] pyrimidin-4-yl] oxy] piperidine-1- Carboxylic acid isopropyl ester (WO2005121121) or (S) -1-methylcyclopropyl 4- (1-fluoro-2- (2- (2,3,6-trifluorophenyl) acetamido) ethyl) Percentage of maximum activity of a test compound relative to that of piperidine-1-carboxylate (see figure below).

^** EC50 was calculated by extrapolating the curve to 100%.

Note that blanks indicate that the test was not performed for that example.

(S) -1-Methylcyclopropyl 4- (1-fluoro-2- (2- (2,3,6-trifluorophenyl) acetamido) -ethyl) piperidine-1-carboxylate ^*** structure

^*** 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 presented by 'by Kathleen Farley.'

In vivo data

All in vivo protocols were approved by Pfizer's Animal Welfare Committee. Naive male Wistar rats (weight 200-250 g on receipt) were obtained from Harlan Laboratories (Indianapolis, Indiana) and paired in plastic gauges hanging on Sani-Chips sawdust bedding. And Purina 5001 chow freely supplied. Rats were housed under controlled light periods (light cycle 6 am to 6 pm) at controlled temperature and humidity conditions. Rats were adapted to the facility at least one week before the study.

Compound manufacturing

Example 50 was formulated as 10% SDD in vehicle 20 mM Tris buffer (pH 7.4) with 0.5% methylcellulose and 0.5% HPMCAS-HF. A dose (75 mg / kg) was formulated at 15 mg / mL for administration at 5 mL / kg, the required bulk was added to the mortar, ground with a small amount of vehicle into a pestle with a pestle and the additional vehicle mixed When added until this flow and transferred to a stirring vessel, the mortar was rinsed several times with the remaining amount of vehicle and capped to prevent evaporation. Compounds were formulated on the day of dosing and continued stirring with a magnetic stir bar before or during the dosing procedure.

Oral glucose tolerance test (OGTT) protocol

Rats had a vehicle of 90 or 30 minutes before glucose (20 mM Tris buffer (pH 7.4) with 0.5% methylcellulose and 0.5% hydroxypropyl methylcellulose acetate succinate-high grade, fine particles (HPMCAS-HF)), or 75 mg / kg of glucose for 90 minutes or 30 minutes before glucose stratified into one of the four dose groups of Example 50 (n = 8 / group). Stratification was performed according to body weight on Day-1, so that each group had the same group mean weight value. Rats were fasted overnight (~ 15 hours) in clean cages before oral glucose tolerance test. Body weights were recorded in the morning of study day (after fasting) for dose volume calculations. Blood samples were collected from all rats through the tail vein and then vehicle or test compound was administered via oral gavage (5 mL / kg). After 90 or 30 minutes rats were bled and immediately administered an oral dose of glucose (2 g / kg). Blood was collected again in rats at 15, 30, 60 and 120 minutes after glucose loading. Blood samples (˜250 μL / time) were collected into EDTA tubes with aprotinin / DPPIVi (0.6 μL per mL of whole blood). Blood tubes were inverted several times immediately after collection, placed on ice and then spun for 5 minutes at 14,000 rpm in a refrigerated centrifuge. Plasma samples were analyzed for glucose levels using the Roche c311 clinical chemistry analyzer, plasma insulin concentrations were determined using Alpco super-sensitive insulin rat ELISA, and total amide GLP-1 concentration was determined by MSD. Determination was made using an ELISA kit.

The results are given as mean +/- SEM (standard error of the mean) unless stated otherwise. Statistical evaluation of the data was performed using one-way analysis of variance (ANOVA) with appropriate post analysis between time-matched vehicle and treatment groups. The difference compared to vehicle with P <0.05 is considered to be statistically significant using the unadjusted T-test.

Table 1:

Preparation of starting material

Preparation Example 1 Isopropyl 4-hydrazinopiperidine-1-carboxylate dihydrochloride salt

Isopropyl 4- {2- (tert-butoxycarbonyl) hydrazinyl} piperidine-1-carboxylate (20.2 g, 67.02 mmol) (obtained as described in WO2008137436) was dissolved in anhydrous ethanol (250 mL) Dissolved in, 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 for 4 h at room temperature under nitrogen. The reaction was concentrated to a white solid containing 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 at rt for 14 h. The reaction was then concentrated under reduced pressure to afford a white solid, which was treated with heptane (100 mL) and concentrated again to give the title compound (15 g, 81%) as a white solid.

Preparation Example 2 isopropyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -piperidine-1-carboxylate

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) were stirred at 85 ° C. for 3 hours. The mixture was concentrated to about one 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 was concentrated in vacuo to afford the crude title compound (9.8 g) as a light yellow solid which was used for the next step without further purification. Analytical samples were prepared by purification via chromatography on silica gel eluting with 30% to 60% ethyl acetate solution in heptane.

Preparation Example 3: Isopropyl 4- [5-bromo-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

Pure tert-butyl nitrite (4.8 mL, 39.3 mmol) isopropyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] in acetonitrile (100 mL) at room temperature. -Added slowly to a stirred mixture of piperidine-1-carboxylate (Preparation Example 2) (8.5 g, 26.2 mmol) and copper (II) bromide (3.7 g, 16 mmol). A significant exothermic effect was observed when the mixture was warmed to about 50 ° C. After continuing heating at 65 ° C. for 30 minutes, the reaction was cooled to room temperature and then concentrated in vacuo. Excess 10% aqueous ammonia was added and the mixture was extracted with ethyl acetate. The organic phase was washed with water and brine and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with 30% to 70% ethyl acetate in heptane to afford the title compound as a yellow oil, which was about 70% pure by NMR and LCMS. The material was used for next step without further purification.

Preparation Example 4 isopropyl 4- [5-bromo-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

Isopropyl 4- [5-bromo-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate in tetrahydrofuran (32 mL) cooled to 0 ° C. To a solution of (3.59 g, 6.5 mmol) was added a solution of 2 M 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 at room temperature for 4 hours. The mixture was cooled to 0 ° C. and methanol was added. The resulting solution was allowed to warm to room temperature and stirred for 10 minutes. The solution was recooled to 0 ° C. and 2 M aqueous 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 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 was concentrated in vacuo. Chromatography on silica gel eluting with 55% to 70% ethyl acetate in heptanes afforded the title compound as an oil.

Preparation Example 5 isopropyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

Isopropyl 4- [5-bromo-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate (1.42 g, 4.10 mmol), tris- (dibenzylidene Acetone) dipaladium (156 mg, 0.170 mmol), 1-1'-bis- (diphenylphosphino) ferrocene (192 mg, 0.346 mmol), zinc powder (68.8 mg, 1.06 mmol), zinc cyanide (497 mg, 4.23 mmol) and N, N-dimethylacetamide (20 mL) were mixed in the 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 Florisil ™ pad, diluted with ethyl acetate and water was added. The aqueous phase was extracted three times with ethyl acetate and the combined organic layers were dried over magnesium sulfate. The mixture was filtered and the filtrate was evaporated in vacuo. Chromatography on silica gel eluting with 55% to 70% ethyl acetate in heptanes gave the title compound (1.06 g, 88%) as a green oil, which solidified on standing.

Preparation Example 6 2-Fluoro-4-[(2-hydroxyethyl) thio] phenol

9,9-dimethyl- in 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) 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) Was added and the dark brown reaction solution was heated at 110 ° C. for 16 h. The reaction was allowed to cool to rt, diluted with water and extracted four times with ethyl acetate. The organic extracts were combined and dried over magnesium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a brown oil which was purified by chromatography on silicon gel to give the title compound (985 mg, 76%) as a brown solid.

Preparation Example 7 4-[(2-{[tert-butyl (dimethyl) silyl] oxy} ethyl) thio] -2-fluorophenol

To a solution of 2-fluoro-4-[(2-hydroxyethyl) thio] phenol (985 mg, 5.24 mmol) and imidazole (371 mg, 5.30 mmol) in N, N-dimethylformamide (5 mL). tert-butyldimethylsilyl chloride (814 mg, 5.24 mmol) was added in small portions and the reaction was stirred at rt for 4 h. The reaction was concentrated under reduced pressure and the residue was diluted with water and then extracted three times with ethyl acetate. The combined organic 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 title compound (1.43 g, 90%) as an orange oil, which was used without further purification.

Preparation Example 8 1- [4- (benzyloxy) -3-fluorophenyl] -1 H-tetrazole

To a suspension of 4- (benzyloxy) -3-fluoroaniline (1.04 g, 4.8 mmol) (WO 2005030140) under nitrogen atmosphere, acetic acid (2.3 mL, 38.3 mmol), triethylorthoformate (2.44 mL, 14.4 mmol) And sodium azide (0.34 g, 5.3 mmol) was added and the reaction mixture was heated at 95 ° C. for 2.5 h. The solution was then allowed to cool to rt, water was added, and then extracted three times with ethyl acetate. The extracts were combined, washed with brine and dried over magnesium sulfate. The mixture was filtered, concentrated under reduced pressure and the crude material was purified by chromatography on silicon gel (20-40% ethyl acetate in heptane) to give the title compound (1.12 g, 86%) as a white solid.

Preparation Example 9 2-Fluoro-4- (1H-tetrazol-1-yl) phenol

To 1- [4- (benzyloxy) -3-fluorophenyl] -1 H-tetrazole (1.12 g, 4.14 mmol) in a Par shake flask was added ethanol (40 mL) and the solution was purged with nitrogen gas. 10% palladium on carbon (0.30 g) was added and the reaction was hydrogenated on a Parr shake machine at 40 psi of hydrogen for 30 minutes. The mixture was then filtered through a fine pore filter and the filtrate was concentrated under reduced pressure to give the title compound (0.67 g, 90%) as a white solid, which was used without further purification.

Preparation Example 10 Isopropyl 4- (5-cyano-4-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

Isopropyl 4- (5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (Preparation 5) (75 mg, 0.24 mmol) was anhydrous. Dissolve in 1 mL dichloromethane and add triethylamine (0.1 mL, 0.74 mmol). After the reaction mixture was cooled in an ice bath, methanesulfonic anhydride (62 mg, 0.34 mmol) was 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 separated. The aqueous layer was extracted more than three times with dichloromethane. The organic extracts were combined, washed with brine, dried over sodium sulfate, filtered and the filtrate was concentrated to give an oil (75 mg, yield 100%). The crude material was used in the next step without further purification.

Preparation Example 11 tert-butyl 4-hydrazinopiperidine-1-carboxylate hydrochloride salt

Hydrazine mono-hydrochloride (17.2 g, 0.25) in water (100 mL) to a solution of tert-butyl 4-oxopiperidine-1-carboxylate (50 g, 0.25 mmol) in methanol (500 mL) in an autoclave mmol) was added. The white mixture was stirred under argon and then 5% palladium on carbon (750 mg) as slurry in water was added. The autoclave was sealed, charged to 60 atmospheres with hydrogen and the reaction stirred for 15 hours. After completion, the reaction was filtered through Celite ^® and the pad was 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, Wash several times with water. The aqueous filtrate was then concentrated under reduced pressure to afford the desired product (221 g, 59%) as a colorless solid.

Preparation Example 12 tert-Butyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

tert-butyl 4-hydrazinopiperidine-1-carboxylate hydrochloride salt (221 g, 880 mmol), ethyl 2-cyano-3-ethoxyacrylate (153 g, 880 mmol), sodium acetate 3 A mixture of hydrate (477 g, 352 mmol) and ethanol (2000 mL) was stirred at 85 ° C. for 8 hours. The mixture was concentrated under reduced pressure and the residue was dissolved in ethyl acetate and water. The layers were separated and the aqueous layer was extracted with ethyl acetate. The combined organic extracts were then dried over magnesium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure. The crude material was purified by filtration through a short plug of silica gel eluting with 40% ethyl acetate in heptanes to give the product as light yellow solid (214 g, 72%).

Preparation Example 13: tert-Butyl 4- [5-bromo-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

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 ° C. Herein tert-butyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate (215 g, in acetonitrile (650 mL) 640 mmol) was added dropwise over 30 minutes. After 4 hours, the reaction was allowed to cool to room temperature and then poured into 2 M hydrochloric acid (1500 mL) on ice. The mixture was extracted three times with ethyl acetate 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 was concentrated under reduced pressure. The resulting residue is purified by filtration through a short silica gel plug eluting with 10% heptane in dichloromethane and then with dichloromethane to give the title compound (137 g, 53%) as a yellow oil, which solidifies on standing. It became.

Preparation Example 14 tert-Butyl 4- [5-bromo-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

Tert-butyl 4- [5-bromo-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxyl in tetrahydrofuran (1300 mL) cooled to 0 ° C. To the solution of rate (137 g, 0.34 mol) 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) was added dropwise. The solution was then stirred for 20 minutes at room temperature. 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 organic layers were washed with brine, dried over magnesium sulfate and the solvent was removed under reduced pressure. The resulting residue was purified by filtration through a short silica gel plug eluting with 30% ethyl acetate in heptane to afford the title compound (61.4 g, 50%) as a colorless oil. The impurity material from the above purification was further purified via the above chromatography procedure to give the second batch of title compound (22 g, 18%) as a colorless solid.

Preparation Example 15 tert-Butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

Copper cyanide (I) (2.97 g, 33.3 mmol) was removed in tert-butyl 4- [5-bromo-4- (hydroxymethyl) -1H-pyrazol-1-yl in degassed dimethylformamide (100 mL). ] Added to a stirred solution of piperidine-1-carboxylate (10 g, 27.8 mmol). The reaction was then heated at 165 ° C. for 4 hours and allowed to cool to room temperature. It was further cooled in an ice bath, a solution of ethylenediamine (5.5 mL) in water (20 mL) was added and then diluted with additional 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 was concentrated under reduced pressure. The procedure was performed in eight batches. The final crude residue was combined and purified by repeated silica gel column chromatography eluting with 40% ethyl acetate in heptanes to give the title compound (11.6 g, 17%) as a colorless solid.

See Example 50 for an alternative synthesis of tert-butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate.

Preparation Example 16: tert-Butyl 4- (5-cyano-4-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

Tert-butyl 4- (5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (202 mg, 0.659 mmol in dichloromethane (6.6 mL) To a stirred solution of) triethylamine (0.18 mL, 1.32 mmol) was added followed by methanesulfonic anhydride (189 mg, 1.1 mmol) at room temperature. After the mixture was stirred for 4.5 h, 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)-as oil. 1H-pyrazol-1-yl) piperidine-1-carboxylate was obtained and 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- (trimethylsilyl) ethoxy) methyl) -2H-tetrazol-5-yl) phenol

A) 4- (benzyloxy) -3-fluorobenzonitrile

To a stirred solution of 3-fluoro-4-hydroxybenzonitrile (1.00 g, 7.30 mmol) in 20 mL of acetonitrile was added small portions of 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 rt for 70 h and then diluted with ethyl acetate and water. The organic phase was separated, 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 5 to 20% gradient of ethyl acetate in heptanes to give 4- (benzyloxy) -3-fluorobenzonitrile (1.33 g) as a white solid.

B) 5- (4- (benzyloxy) -3-fluorophenyl) -1H-tetrazole and 5- (4- (benzyloxy) -3-fluorophenyl) -2H-tetrazole

4- (benzyloxy) -3-fluorobenzonitrile (250 mg, 1.10 mmol), sodium azide (214 mg, 3.30 mmol), ammonium chloride (176 mg, 3.30 mmol) and N, N-dimethylformamide The vial filled with 3 mL was heated at 110 ° C. for 18 h. The reaction mixture was cooled to rt, diluted with water and ethyl acetate and the pH was adjusted to 3 with 1 N aqueous hydrochloric acid. The organic phase was separated, washed with brine, dried over magnesium sulfate, filtered and the filtrate was concentrated in vacuo to afford the title compound (270 mg) as a white solid. The material was used for the next step without further purification.

C) 5- (4- (benzyloxy) -3-fluorophenyl) -1-((2- (trimethylsilyl) ethoxy) methyl) -1H-tetrazole and 5- (4- (benzyloxy)- 3-fluorophenyl) -2-((2- (trimethylsilyl) ethoxy) methyl) -2H-tetrazole

5- (4- (benzyloxy) -3-fluorophenyl) -1H-tetrazole and 5- (4- (benzyloxy) -3-fluorophenyl) -2H-tetrazole (dissolved in tetrahydrofuran) To a solution of 270 mg, 1 mmol) sodium hydride (44 mg, 1.1 mmol) was added in four portions and the resulting mixture was stirred at room temperature for 15 minutes. Then (2- (chloromethoxy) ethyl) trimethylsilane (0.19 mL, 1.0 mmol) was added and the reaction mixture was stirred at rt for 16 h. The reaction was quenched by addition of water and ethyl acetate was added. The organic phase was separated and the aqueous phase 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 heptane (5 to 20% ethyl acetate) gave the desired product (270 mg, yield 67%) as a white solid.

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

5- (4- (benzyloxy) -3-fluorophenyl) -1-((2- (trimethylsilyl) ethoxy) methyl) -1H-tetra dissolved in a mixture of 2 mL ethanol and 2 mL tetrahydrofuran Palladium in a solution of sol and 5- (4- (benzyloxy) -3-fluorophenyl) -2-((2- (trimethylsilyl) ethoxy) methyl) -2H-tetrazole (140 mg, 0.35 mmol) Black (56 mg, 0.53 mmol) and formic acid (0.14 mL, 3.5 mmol) were added. The resulting mixture was stirred at rt for 4 h and then filtered through a Celite ^® pad. The filtrate was concentrated under reduced pressure and the resulting crude was used in the next step without further purification.

Preparation Example 18 5- (4- (benzyloxy) -3-fluorophenyl) -1- (2- (trimethylsilyloxy) ethyl) -1H-tetrazole and 5- (4- (benzyloxy) -3 -Fluorophenyl) -2- (2- (trimethylsilyloxy) ethyl) -2H-tetrazole

5- (4- (benzyloxy) -3-fluorophenyl) -1H-tetrazole and 5- (4- (benzyloxy) -3-fluoro dissolved in N, N-dimethylformamide (8 mL) To a solution of phenyl) -2H-tetrazole (Preparation 17, Step B) (550 mg, 2 mmol) was added sodium hydride (163 mg, 4 mmol) in two portions and the resulting mixture was stirred at room temperature for 5 minutes. Stirred. Then (2-bromoethoxy) trimethylsilane (1.3 mL, 6 mmol) was added and the reaction mixture was stirred at 70 ° C. for 16 h and then cooled to rt. The reaction was quenched by addition of water and ethyl acetate was added. The organic phase was separated and the aqueous phase 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-fluorophenyl) -1- (2- ( Trimethylsilyloxy) ethyl) -1H-tetrazole (100 mg, yield 12%) and 5- (4- (benzyloxy) -3-fluorophenyl) -2- (2- (trimethylsilyloxy) ethyl)- 2H-tetrazole (600 mg, 69% yield) was obtained.

Preparation 19: 2-Fluoro-4- (2- (2- (trimethylsilyloxy) ethyl) -2H-tetrazol-5-yl) phenol

5- (4- (benzyloxy) -3-fluorophenyl) -2- (2- (trimethylsilyloxy) ethyl) -2H-tetrazole (prepared in a mixture of 6 mL ethanol and 6 mL tetrahydrofuran) Example 18) (230 mg, 0.54 mmol) was added palladium black (86 mg, 0.806 mmol) and formic acid (0.215 mL, 5.4 mmol). The resulting mixture was stirred at rt for 4 h and then filtered through a Celite ^® pad. The filtrate was concentrated under reduced pressure and the resulting crude material (180 mg) was used in the next step without further purification.

Preparation Example 20 2-Fluoro-4- (1- (2- (trimethylsilyloxy) ethyl) -1H-tetrazol-5-yl) phenol

5- (4- (benzyloxy) -3-fluorophenyl) -1- (2- (trimethylsilyloxy) ethyl) -1H-tetrazole (prepared in a mixture of 2 mL ethanol and 2 mL tetrahydrofuran) Example 18) To a solution of (130 mg, 0.30 mmol) was added palladium black (48 mg, 0.45 mmol) and formic acid (0.12 mL, 3 mmol). The resulting mixture was stirred at rt for 4 h and then filtered over a Celite ^® pad. The filtrate was concentrated under reduced pressure and the resulting crude material (94 mg) was used in the next step without further purification.

Preparation Example 21 2-Fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenol

A) 5- (4- (benzyloxy) -3-fluorophenyl) -1-methyl-1H-tetrazole and 5- (4- (benzyloxy) -3-fluorophenyl) -2-methyl-2H Tetrazole

5- (4- (benzyloxy) -3-fluorophenyl) -1H-tetrazole and 5- (4- (benzyloxy) -3-fluorophenyl) -2H-tetrazole in 30 mL of tetrahydrofuran ( To a stirred solution of Preparation 17, step B) (1.50 g, 5.55 mmol), sodium hydride (444 mg, 11.1 mmol) was added in two portions at room temperature. After 5 minutes, iodomethane (1.04 mL, 16.6 mmol) was added and the reaction stirred at room temperature under nitrogen atmosphere for 15 hours. The mixture was diluted with water and 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 chromatography eluting with 10-40% ethyl acetate in heptanes to give 5- (4- (benzyloxy) -3-fluorophenyl) -2-methyl-2H-tetrazole as a white solid ( 1.1 g) and 5- (4- (benzyloxy) -3-fluorophenyl) -1-methyl-1H-tetrazole (450 mg) as a white solid were obtained.

5- (4- (benzyloxy) -3-fluorophenyl) -1-methyl-1H-tetrazole.

B) 2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenol

In a solution of 5- (4- (benzyloxy) -3-fluorophenyl) -1-methyl-1H-tetrazole (500 mg, 1.76 mmol) in 6 mL of ethanol and 6 mL of tetrahydrofuran, formic acid (0.7 mL, 17.6 mmol) followed by palladium black (281 mg, 2.64 mmol). The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was filtered through Celite ^® . The filtrate was concentrated in vacuo to afford 2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenol (330 mg) as a white solid which was used in the next step without further purification. .

Preparation Example 22 4- (1-Methyl-1H-tetrazol-5-yl) phenol

A) 4- (benzyloxy) -N-methylbenzamide

In a flask filled with thionyl chloride (0.35 mL, 4.82 mmol) at 0 ° C., 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 The solution was added with stirring. The ice bath was removed and the solution was stirred at rt for 4 h. The mixture was concentrated in vacuo to yield a white solid. The solid was dissolved 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, the organic layer was separated, dried over magnesium sulfate, filtered and the filtrate was concentrated in vacuo to give a white solid. The solid was recrystallized from ethyl acetate and heptane to give 4- (benzyloxy) -N-methylbenzamide (850 mg) as a white needle.

B) 5- (4- (benzyloxy) phenyl) -1-methyl-1H-tetrazole

In a flask equipped with a reflux condenser in a stirred solution of 4- (benzyloxy) -N-methylbenzamide (200 mg, 0.829 mmol) in 3 mL of acetonitrile and 1 drop of N, N-dimethylformamide, Ethylamine (0.12 mL) was added.

The reaction mixture was stirred for 10 minutes, then thionyl chloride (0.078 mL, 1.08 mmol) was added dropwise. 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 stirred vigorously 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 10 to 40% gradient ethyl acetate in heptanes to give 5- (4- (benzyloxy) phenyl) -1-methyl-1H-tetrazole (180 mg) as a white solid. Obtained.

C) 4- (1-methyl-1H-tetrazol-5-yl) phenol

Formic acid (0.27 mL, 6.76 mmol) in a stirred solution of 5- (4- (benzyloxy) phenyl) -1-methyl-1H-tetrazole (180 mg, 0.676 mmol) in 3 mL of ethanol and 3 mL of tetrahydrofuran. Then palladium black (108 mg, 1.01 mmol) was added. 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 (110 mg) as a white solid, which was followed without further purification. Used for reaction.

Preparation Example 23 3-Fluoro-4-hydroxybenzamide

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 h. After cooling to rt, the mixture was concentrated in vacuo, the residue was taken up in water, acidified with acetic acid and extracted with ethyl acetate. The combined organic extracts were dried over magnesium sulphate, filtered and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography eluting with 20 to 60% gradient ethyl acetate in heptanes to afford 3-fluoro-4-hydroxybenzamide (210 mg) as a white solid.

Alternatively, 3-fluoro-4-hydroxybenzamide can be prepared as follows:

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 and then a solution of 3-fluoro-4-hydroxybenzonitrile (1.00 g, 7.29 mmol) in 5 mL of ethanol was added. The mixture was stirred vigorously for 2 hours at room temperature and then diluted with water (100 mL) and ethyl acetate (100 mL). After the mixture was stirred for 5 minutes, 1 M hydrochloric acid was added to pH 4. The aqueous layer was separated and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over magnesium sulphate, filtered and the filtrate was concentrated to give a white solid. The solid was triturated with diethyl ether and heptane (2: 1, 90 mL) for 1 hour and then filtered to afford 3-fluoro-4-hydroxybenzamide (1.05 g) as a white solid.

Preparation Example 24 2-Fluoro-4-hydroxybenzamide

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 and then a solution of 2-fluoro-4-hydroxybenzonitrile (500 mg, 3.65 mmol) in 2 mL of ethanol was added. The mixture was stirred vigorously for 2 hours at room temperature and then diluted with water (100 mL) and ethyl acetate (100 mL). After the mixture was stirred for 5 minutes, 1 M hydrochloric acid was added to pH = 4. The aqueous layer was separated and extracted with ethyl acetate (3X50 mL). The combined organic layers were dried over magnesium sulphate, filtered and the filtrate was concentrated to give 2-fluoro-4-hydroxybenzamide as a white solid.

Preparation Example 25 Isopropyl 4- (5-cyano-4- (1-hydroxyethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

Isopropyl 4- (5-cyano-4-formyl-1H-pyrazol-1-yl) piperidine-1-carboxylate (Example 9, Step A) (51 mg, 0.18 mmol) It was dissolved in 2 mL of tetrahydrofuran and cooled to -78 ° C under nitrogen atmosphere. Then methylmagnesium bromide (0.070 mL, 0.21 mmol, 3 M in diethyl ether) was added dropwise. The cold bath was removed and the mixture was stirred at rt for 1 h. 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 yield isopropyl 4- (5-cyano-4- (1-hydroxyethyl)-as a white solid. 1H-pyrazol-1-yl) piperidine-1-carboxylate (33 mg) was obtained and used in the next step without further purification.

Preparation 26: 1-Methylcyclopropyl 4-nitrophenyl carbonate

A) 1-methylcyclopropanol

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 ° C. until methanol was removed. Toluene was removed at 180 ° C. Additional toluene was added and the process repeated twice. After all 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. The 5 L three-necked flask was equipped with an overhead stirrer, an inert gas inlet and a pressure-equilibration 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 75.6 mL ether) and diethyl ether (1500 mL). The solution was stirred while maintaining the reaction flask in a room temperature water bath. The addition funnel was charged with 3 M ethylmagnesium bromide solution (554 mL, 1.66 moles). Grignard reagent was added dropwise over 3 hours at room temperature. The mixture became a light yellow solution, then gradually a precipitate formed, eventually turning to a dark green / brown / black mixture. After stirring for an additional 15 minutes, after the Grignard was added, 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 solids dissolved. The aqueous layer was separated and extracted with 2 x 500 mL of diethyl ether. 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 fractions boiling at about 100 ° C. while distilling off the crude were collected to give the title compound (23 g), methyl ethyl ketone and 2-butanol as small amounts of impurities.

The preparation of the title compound is also described in WO09105717.

B) 1-methylcyclopropyl 4-nitrophenyl carbonate

1-methylcyclopropanol (10 g, 137 mmol), 4-nitrophenyl chloroformate (32 g, 152 mmol) and 4-dimethylaminopyridine (150 mg, 1.2 mmol) in minor crystals in dichloromethane (462 mL) The solution of was cooled to 0 ° C. Triethylamine (36.5 g, 361 mmol) was added dropwise. 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 sulphate, filtered and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography, eluting with a gradient of an ethyl acetate mixture in heptane (0 to 5% ethyl acetate over 10 minutes, then heptane in an isocratic of 5% ethyl acetate) to give the desired carbo as a clear oil. 20.8 g of Nate were obtained. This oil solidified upon standing.

Alternatively, 1-methylcyclopropanol can be prepared as follows:

1-methylcyclopropanol

The 2000 mL four-necked flask was equipped with a mechanical stirrer, inert gas inlet, thermometer and two pressure-equilibration 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. A 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 below 10 ° C. 40 mL of methyl acetate solution was added to the flask. The Grignard reagent was then added dropwise from the addition funnel at a rate no faster than 2 mL per minute dropping about 2 drops per second. After 40 mL of Grignard reagent first was added, an additional 20 mL portion of methyl acetate in ether solution was added. After 40 mL of Grignard Reagent 2 was added, an additional 20 mL portion of methyl acetate in diethyl ether solution was added. After 40 mL of Grignard reagent 3 was added, an additional 20 mL portion of methyl acetate in ether solution was added. After 40 mL of Grignard Reagent 4 was added, the last 20 mL portion of methyl acetate in ether solution was added.

The mixture was stirred for an additional 15 minutes after completion of Grignard reagent addition. The mixture was then stirred rapidly while pouring into a mixture of 660 g ice and 60 mL concentrated sulfuric acid 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 g of 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 transfer. Distillation at atmospheric pressure was continued through a 2 cm x 20 cm jacketed Vigreux column. The liquid distilled at 98-105 ° C. was collected to give 14 g of the title compound as a colorless liquid.

Preparation 27: 2-Fluoro-4- (1-methyl-1H-imidazol-5-yl) phenol

A) 5- (3-fluoro-4-methoxyphenyl) -1-methyl-1H imidazole

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 an additional 10 minutes. . The vessel was placed in a microwave reactor at 140 ° C. for 2 hours. The mixture was diluted with ethyl acetate, filtered through Celite ^® and the filtrate was concentrated under reduced pressure. The crude was purified by chromatography eluting with a gradient of 25 to 100% ethyl acetate in heptane followed by 0 to 10% methanol in dichloromethane to afford the title compound (210 mg) as a yellow oil.

Proton migration at 7.42 is an indicator of the desired imidazole isomer compared to the literature (Eur. J. Org. Chem., 2008, 5436 and Eur. J. Org., 2006, 1379).

B) 2-fluoro-4- (1-methyl-1H-imidazol-5-yl) phenol

Boron bromide (III) solution (0.50 mL, in a solution of 5- (3-fluoro-4-methoxyphenyl) -1-methyl-1H imidazole (101.8 mg, 0.494 mmol) in dichloromethane (2.0 mL) 1.0 M solution in heptane) was added at -30 ° C. The mixture was stirred at room temperature for 20 hours. The mixture was then cooled to -30 ° C and methanol (2 mL) was added to the mixture. The mixture was concentrated in vacuo and the residue dissolved in water and neutralized with 1 M sodium hydroxide. The solution was concentrated to give the title compound (90 mg) as a yellow solid. This compound was used without further purification.

Preparation Example 28 2-Fluoro-4- (1-methyl-1H-imidazol-2-yl) phenol

A) 2- (3-fluoro-4-methoxyphenyl) -1-methyl-1H imidazole

2-fluoro-4-bromoanisole (0.256 mL, 1.93 mmol) and copper iodide (I) (375 mg, 1.93 mmol) were placed in microwave vials and N, N-dimethylformamide (4.8 mL). )). The mixture was degassed with a stream of nitrogen gas for 10 minutes, 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 an additional 10 minutes. . The vessel was placed in a microwave reactor at 140 ° C. for 2 hours. The mixture was diluted with ethyl acetate (3 mL), poured into saturated aqueous ammonium chloride solution, stirred for 30 minutes in open air, which was extracted twice with ethyl acetate. The combined organic phases were washed with water, dried over sodium sulphate, filtered and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography eluting with a gradient of ethyl acetate mixture in heptane (25 to 100% ethyl acetate / heptane, then 0 to 10% methanol in dichloromethane) to give 2- (3-fluoro as a yellow oil. 4-methoxyphenyl) -1-methyl-1H imidazole (35.8 mg) was obtained.

Proton NMR is characterized by proton NMR of 5- (3-fluoro-4-methoxyphenyl) -1-methyl-1H imidazole (Preparation 27) and Eur. J. Org. chem., 2008, 5436 and Eur. J. Org., 2006, 1379]), which represents the desired imidazole isomer.

B) 2-fluoro-4- (1-methyl-1H-imidazol-2-yl) phenol

2-Fluoro-4- (1-methyl-1H-imidazol-2-yl) phenol was prepared according to a procedure similar to the method of Preparation 27 (B). Preparation from -1-methyl-1H imidazole gave the title compound (33.4 mg) as a brown solid. The crude material was used without further purification.

Preparation 29: 2-Fluoro-4- (methylsulfonyl) -1- (prop-1-en-2-yl) benzene

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) To the solution was sequentially added the catalyst 1,1'-bis- (diphenylphosphino) -ferrocene palladium dichloride (67 mg, 0.089 mmol) and triethylamine (0.17 mL, 1.20 mmol). The reaction was heated at 90 ° C. 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 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 was concentrated under reduced pressure. The residue was purified by silica gel chromatography (10 to 100% ethyl acetate in heptanes) to give the title compound (130 mg, 80%) as a white solid.

Preparation 30: 4-hydroxy-2-methylbenzonitrile

Boron trichloride in dichloromethane (61.2 mL, 1M) was added slowly to dichloromethane (93 mL) and cooled to -78 ° C. 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 ° C for 5 minutes. The reaction mixture was then slowly warmed to room temperature and stirred for 2 hours. The reaction was quenched by the slow addition of ice slurry. The reaction was allowed to stir for 30 minutes and the layers separated. The aqueous layer was extracted with dichloromethane (2x) and the combined organic extracts passed through a phase separation 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 (1.85 g, 68%) as a yellow solid.

Preparation 31A: 3-Fluoro-4-hydroxy-N-methylbenzamide

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 h. The solution was cooled to rt, the solid was filtered off 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 afford the desired product benzyl 4- (benzyloxy) -3-fluorobenzoate.

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 a 1 M aqueous solution of hydrochloric acid. The white solid was precipitated, filtered and dried overnight to afford the desired product, 4- (benzyloxy) -3-fluorobenzoic acid (7.6 g, 90%) as a white solid.

C) 4- (benzyloxy) -3-fluoro-N-methylbenzamide

Thionyl chloride (2.7 mL, 37 mmol) is added to a solution of 4- (benzyloxy) -3-fluorobenzoic acid in dimethylformamide (0.048 mL, 0.617 mmol) and dichloromethane (100 mL) at 0 ° C., The resulting solution was stirred at rt for 20 h. 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), a 2 M solution of methylamine in tetrahydrofuran (35 mL) was added, and the reaction was stirred at rt for 18 h. The reaction mixture was concentrated to half of original volume under reduced pressure, and a white solid precipitated out. The solid was filtered off, washed with water and dried overnight in a vacuum oven to afford the desired product (6.00 g, 70%) as a white solid.

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 constant stream of nitrogen. The reaction was shaken under a 50 psi atmosphere of hydrogen for 64 hours at room temperature. The reaction mixture was carefully filtered through a pad of Celite ^® while washing with excess ethyl acetate. The filtrate was concentrated under reduced pressure to afford the desired product (628 mg, 93%) as a light brownish yellow solid.

Preparation 31B: 3-Fluoro-4-hydroxy-N, N-dimethylbenzamide

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 diisopropyl Ethyl amine (4.5 mL, 26 mmol) was 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 at room temperature overnight. The reaction was diluted with dichloromethane and 1 M phosphoric acid. The precipitate 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%).

Preparation Example 32 tert-Butyl 3-hydroxy-4,4-dimethoxypiperidine-1-carboxylate

tert-butyl 4-oxo-1-piperidinecarboxylate (2.00 g, 10 mmol) was dissolved in methanol (20 mL) and cooled to 0 ° C. Powdered potassium hydroxide (1.26 g, 22.1 mmol) was added. Iodine (2.8 g, 11 mmol) was dissolved in methanol (25 mL) and added dropwise to the reaction over 45 minutes. The reaction was then slowly warmed to rt and stirred for 16 h. The reaction was concentrated and toluene (50 mL) was added. The resulting solid was 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 of 30% to 100% ethyl acetate in heptane and tert-butyl 3-hydroxy-4,4-dimethoxypiperidine- 1-carboxylate (1.89 g, 72%) was obtained.

Preparation Example 33 tert-Butyl 3-hydroxy-4-oxopiperidine-1-carboxylate

tert-butyl 3-hydroxy-4,4-dimethoxypiperidine-1-carboxylate (6.70 g, 26 mmol) is dissolved in acetone (135 mL) and p-toluene sulfonic acid (244 mg, 1.28 mmol) ) Was added. The reaction was stirred at rt for 16 h. 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-oxopiperidine-1-carboxylate (4.67 g, 69%) as an oil. .

Preparation Example 34 tert-butyl 4-hydrazino-3-hydroxypiperidine-1-carboxylate

tert-butyl 3-hydroxy-4-oxopiperidine-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, which was also added to the reaction. 10% platinum catalyst on carbon (500 mg) was slurried in water and added to the reaction mixture. The mixture was shaken under a 50 psi atmosphere of hydrogen at room temperature for 16 hours. The reaction was filtered through a pad of Celite ^® while washing with methanol. The filtrate was concentrated under reduced pressure, then diluted with heptane and concentrated under reduced pressure to afford the desired product, tert-butyl 4-hydrazino-3-hydroxypiperidine-1-carboxylate.

Preparation Example 35 tert-Butyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-hydroxypiperidine-1-carboxylate

tert-butyl 4-hydrazino-3-hydroxypiperidine-1-carboxylate (5.30 g, 20 mmol) and ethyl (ethoxymethylene) cyanoacetate (3.42 g, 19.8 mmol) were dissolved in anhydrous 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 rt, concentrated under reduced pressure and the resulting residue diluted with water and ethyl acetate. The organic layer was dried over sodium sulphate, filtered and the filtrate was concentrated under reduced pressure. The crude oil was purified by flash chromatography eluting with a gradient of 10% to 100% ethyl acetate in heptane, tert-butyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazole-1- Il] -3-hydroxypiperidine-1-carboxylate was obtained.

Preparation Example 36 tert-Butyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate

tert-butyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-hydroxypiperidine-1-carboxylate (1.71 g, 4.82 mmol) It was dissolved in dichloromethane (50 mL) and cooled to -78 ° C. Diethylaminosulfur trifluoride (0.710 mL, 0.58 mmol) was added dropwise and then warmed to 0 ° C. for 25 minutes. The reaction solution was cooled to -78 ° C and methanol (10 mL) was added carefully. The reaction was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with a gradient of 10% to 100% ethyl acetate in heptane to give tert-butyl 4- [5-amino-4- (ethoxycarbonyl)- 1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate was obtained.

Preparation Example 37 tert-Butyl 4- [5-bromo-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate

tert-butyl 4- [5-amino-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate (710 mg, 1.99 mmol) It was dissolved in acetonitrile (25 mL). Copper (II) bromide (539 mg, 2.39 mmol) was added and the reaction was heated to 60 ° C. tert-butyl nitrile (0.315 mL, 2.9 mmol) was added dropwise and the mixture was heated at 65 ° C. for 15 minutes. The reaction was cooled to rt, poured into cold 1 N hydrochloric acid and extracted with ethyl acetate (2 ×). The combined organic extracts were washed with saturated aqueous sodium bicarbonate and brine, 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 of 10% to 50% ethyl acetate in heptane, tert-butyl 4- [5-bromo-4- (ethoxycarbonyl) -1H-pyrazole- 1-yl] -3-fluoropiperidine-1-carboxylate (320 mg, 38%) was obtained.

Preparation Example 38 Ethyl 5-cyano-1- (3-fluoropiperidin-4-yl) -1H-pyrazole-4-carboxylate

tert-butyl 4- [5-bromo-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate (185 mg, 0.31 mmol) , 1,1'bis (diphenylphosphino) ferrocene (18 mg, 0.032 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 ° C. for 4.5 hours. The reaction mixture was cooled to rt 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 followed by brine, 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 of 10% to 100% ethyl acetate in heptanes to afford ethyl 5-cyano-1- (3-fluoropiperidin-4-yl) -1H-pyra. Obtained sol-4-carboxylate (80 mg, 98%).

Preparation Example 39 tert-butyl 4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate

Ethyl 5-cyano-1- (3-fluoropiperidin-4-yl) -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 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 of 10% to 100% ethyl acetate in heptanes to give tert-butyl 4- [5-cyano-4- (ethoxycarb as oil). Bonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate (52 mg, 63%) was obtained.

Preparation 40: 1- [1- (tert-butoxycarbonyl) -3-fluoropiperidin-4-yl] -5-cyano-1H-pyrazole-4-carboxylic acid

tert-butyl 4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -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 ° C. and lithium hydroxide monohydrate (19 mg, 0.436 mmol) was added. The reaction was allowed to warm slowly to room temperature over 2.5 hours. Concentrate the reaction mixture; 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 to pH 2 with 1 N aqueous sodium bisulfate. The acidic solution is extracted with ethyl acetate (3x), the extract is washed with brine, dried over sodium sulfate, filtered and the filtrate is concentrated under reduced pressure to give 1- [1- (tert-butoxycarbonyl) as a white solid. -3-fluoropiperidin-4-yl] -5-cyano-1H-pyrazole-4-carboxylic acid was obtained.

Preparation Example 41 tert-Butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate

Freshly recrystallized cyanuric acid chloride (78 mg, 0.414 mmol) (from heptane) was dissolved in dimethoxyethane (2 mL) and 4-methyl-morpholine (0.020 mL, 0.215 mmol) was added. 1- [1- (tert-butoxycarbonyl) -3-fluoropiperidin-4-yl] -5-cyano-1H-pyrazole dissolved in dimethoxyethane (2 mL) in this viscous solution 4-carboxylic acid (70 mg, 0.21 mmol) was added. The reaction was heated at 60 &lt; 0 &gt; C for 3 hours. The reaction was cooled to room temperature and filtered through a pad of Celite ^® while washing with dimethoxyethane. The filtrate was cooled to 0 ° C. and a solution of sodium borohydride (17 mg, 0.474 mmol) dissolved in water (0.4 mL) was added very slowly (dropwise). Once the addition was complete, the reaction was allowed to warm to room temperature for 2.5 hours. The reaction solution was further diluted with water and acidified to pH 2.5 with 1 M 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 gradient of 10% to 100% ethyl acetate in heptanes to give tert-butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazole as an oil. -1-yl] -3-fluoropiperidine-1-carboxylate (28 mg, 42%) was obtained.

Preparation Example 42 tert-Butyl 4- (5-cyano-4-{[(methylsulfonyl) oxy] methyl} -1 H-pyrazol-1-yl) -3-fluoropiperidine-1-car Carboxylate

tert-butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate (28 mg, 0.086 mmol) It was dissolved in dichloromethane (3 mL). Triethylamine (0.036 mL, 0.258 mmol) was added and the mixture was cooled to 0 ° C. Methanesulfonic anhydride (20 mg, 0.112 mmol) was added dropwise and allowed to slowly warm 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 (2 ×) and the combined organic extracts passed through a cotton plug. The filtrate was concentrated under reduced pressure to afford tert-butyl 4- (5-cyano-4-{[(methylsulfonyl) oxy] methyl} -1 H-pyrazol-1-yl) -3-fluoropiperi as oil. Dean-1-carboxylate (33 mg, 95%) was obtained.

Preparation Example 43 Isomers of tert-Butyl-3-fluoro-4-hydroxypiperidine-1-carboxylate (B and C)

The experimental details are described in detail in Scheme 4 below.

Scheme 4

Step A) tert-Butyl-4-[(trimethylsilyl) oxy] -3,6-dihydropyridine-1 (2H) -carboxylate

Trimethylsilyl chloride (22.9 mL, 0.18 mol) and tri in a solution of N-tert-butoxycarbonyl-4-piperidone (30.0 g, 0.15 mol) in dry N, N-dimethylformamide (300 mL) at room temperature Ethylamine (50.4 mL, 0.36 mol) was added sequentially via an addition funnel. The resulting solution was heated at 80 ° C. 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 was concentrated under reduced pressure to afford the crude product as a yellow oil. The oil was purified by passing through a plug of silica gel eluting with 9: 1 heptane / ethyl acetate to afford the title compound (33.6 g, 82%) as a colorless oil.

Step B) tert-Butyl-3-fluoro-4-oxopiperidine-1-carboxylate

To a stirred solution of tert-butyl-4-[(trimethylsilyl) oxy] -3,6-dihydropyridine-1 (2H) -carboxylate (28.8 g, 0.11 mol) in acetonitrile (300 mL) at room temperature Selectfluor ™ (41.4 g, 0.12 mol) was added. The resulting pale yellow suspension was stirred at rt for 1.5 h. Saturated aqueous sodium bicarbonate (300 mL) and ethyl acetate (300 mL) were added and the layers separated. The aqueous layer was extracted twice with ethyl acetate and all organic layers were combined, 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 afford the crude product as pale yellow oil. Purification of this material was repeated column chromatography on silica gel using a heptane / ethyl acetate gradient (2: 1 to 1: 1) to afford the title compound (15.5 g, 67%) as a white solid.

Alternatively step B can be carried out as follows while isolating the hydrate of the ketone.

To a stirred solution of tert-butyl-4-[(trimethylsilyl) oxy] -3,6-dihydropyridine-1 (2H) -carboxylate (41.3 g, 0.15 mol) in acetonitrile (500 mL) at room temperature SelectFluor ™ (56.9 g, 0.16 mol) was added. 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 organic layers were combined, 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 afford crude tert-butyl-3-fluoro-4-oxopiperidine-1-carboxylate as a white solid. The crude tert-butyl-3-fluoro-4-oxopiperidine-1-carboxylate was suspended in tetrahydrofuran (120 mL) and water (120 mL) was added. The resulting solution was stirred at rt for 5.5 h 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 were collected by filtration using a sintered glass funnel. The resulting filter cake was washed thoroughly with a 1: 1 mixture of dichloromethane (200 mL), dichloromethane (200 mL) and heptane (100 mL). The solid was then dried under high vacuum to afford tert-butyl 3-fluoro-4,4-dihydroxypiperidine-1-carboxylate (26.4 g).

Step C) Isomers of (R ^* )-tert-butyl-3- (S) -fluoro-4- (R) -hydroxypiperidine-1-carboxylate (racemic)

Sodium borohydride (3.51 g, 93.7 mmol) in a solution of tert-butyl-3-fluoro-4-oxopiperidine-1-carboxylate (15.5 g, 71.3 mmol) in methanol (150 mL) at 0 ° C. Was added. The resulting mixture was stirred at 0 ° C. 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 is filtered and the filtrate is concentrated under reduced pressure to give a crude product mixture which is purified by column chromatography on silica gel eluting with heptane-ethyl acetate (3: 2-1: 1) as light yellow oil. First elution product, tert-butyl- (3,4-trans) -3-fluoro-4-hydroxypiperidine-1-carboxylate (Compound C, Scheme 4) (3.81 g, 24%) Obtained, which solidified to a white solid upon standing.

Then, a second eluting compound as a white solid, tert-butyl- (3,4-cis) -3-fluoro-4-hydroxy-piperidine-1-carboxylate (Compound B, Scheme 4) (10.57 g, 68%).

Alternatively step C can be carried out starting with hydrate tert-butyl 3-fluoro-4,4-dihydroxypiperidine-1-carboxylate (step 2) according to the following.

Tetrahydrofuran at −35 ° C. in a stirred solution of tert-butyl 3-fluoro-4,4-dihydroxypiperidine-1-carboxylate (20.0 g, 85 mmol) in tetrahydrofuran (500 mL) A solution of L-Selectride ^® in (170 mL, 1 M, 170 mmol) was added dropwise over 30 minutes. The reaction mixture was warmed to 0 ° C. over 1.5 h. The reaction mixture was quenched with saturated aqueous ammonium chloride (150 mL) and stirred vigorously for 15 minutes. To this 0 ° C. mixture was added pH 7 phosphate buffer (150 mL), followed by the dropwise addition of 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 washed sequentially with water, saturated aqueous sodium thiosulfate and brine. The organic layer is then dried over anhydrous magnesium sulphate, filtered and the filtrate is concentrated under reduced pressure to give a crude product mixture which is column chromatography on silica gel eluting with heptane-ethyl acetate (10 to 60% gradient). Purification by [combiflash isco 330 g column] yielded tert-butyl- (3,4-cis) -3-fluoro-4-hydroxypiperidine-1-carboxylate (13.9 g). .

Step D) Enantiomer 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 flown through a mobile phase of 90:10 carbon dioxide and ethanol respectively at a flow rate of 10 Purified to isomer thereof via preparative high pressure liquid chromatography using a Chiralpak AD-H column (10 × 250 mm) using mL / min. The wavelength for monitoring the separation was 210 nm. Analytical purity of each isomer was determined via analytical high pressure chromatography using a Chiralpak AD-H (4.6 mm x 25 cm) column using a mobile phase of 90:10 carbon dioxide and ethanol at a flow rate of 2.5 mL / min. The wavelength for monitoring the peak was 210 nm. Two isomers were obtained:

Compound E, scheme 4) (3S, 4R) -tert-butyl 3-fluoro-4-hydroxypiperidine-1-carboxylate, enantiomer 1 (363 mg): R _t = 2.67 min (100% ee) (light rotation in dichloromethane = +21.2 degrees) and

Compound D, scheme 4) (3R, 4S) -tert-butyl 3-fluoro-4-hydroxypiperidine-1-carboxylate, enantiomer 2 (403 mg): R _t = 2.99 min (88% ee).

Absolute stereochemistry of tert-butyl- (3,4-cis) -3-fluoro-4-hydroxy-piperidine-1-carboxylate isomers was prepared using 5- (6- ((3S, 4R) -3-fluoropiperidin-4-yloxy) -5-methylpyrimidin-4-yl) -1-methyl-1,4,5,6-tetrahydropyrrolo [3 It was determined by preparing (1S)-(+)-camphorsulfonic acid (see similar recipe for racemic form below) of, 4-c] pyrazole.

5- (6-{[(3,4-cis) -3-fluoropiperidin-4-yl] oxy} -5-methylpyrimidin-4-yl) -1-methyl-1,4,5 Preparation of 6-tetrahydropyrrolo [3,4-c] pyrazole (racemic)

A. Preparation of 5- (6-chloro-5-methylpyrimidin-4-yl) -1-methyl-1,4,5,6-tetrahydropyrrolo [3,4-c] pyrazole

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) was suspended in tetrahydrofuran (51 mL) at room temperature. To this was added triethylamine (4.41 mL, 31.6 mmol), which clouded the mixture and gave a brown solid sticking to the flask wall. The mixture was stirred at rt for 4 h and then heated to 50 ° C. for an additional 19 h. The reaction mixture was cooled to rt and diluted with water (100 mL). This mixture was extracted with ethyl acetate (3 x 100 mL). The organic extracts were combined, washed with brine, dried over sodium sulfate and filtered. The filtrate was dried under reduced pressure in vacuo to give the title compound (1.95 g, 78%) as a light brown solid which was used in the next step without further purification.

B. tert-butyl (3,4-cis) -3-fluoro-4-{[5-methyl-6- (1-methyl-4,6-dihydropyrrolo [3,4-c] pyrazole Preparation of -5 (1H) -yl) pyrimidin-4-yl] oxy} piperidine-1-carboxylate (racemic)

Tert-butyl (3,4-cis) -3-fluoro-4-hydroxypiperidine-1-carboxylate (1.67 g, 7.62 mmol) and 5- (6-chloro-5-methyl) A mixture of pyrimidin-4-yl) -1-methyl-1,4,5,6-tetrahydropyrrolo [3,4-c] pyrazole (900 mg, 3.60 mmol) was prepared with 1,4-dioxane ( 20 mL) and heated to 105 ° C. After heating for 10 minutes all the material is in solution and sodium bis (trimethylsilyl) amide (4.3 mL, 4.3 mmol, 1M in toluene) is added quickly to the mixture to give a cloudy yellow mixture which is then 2 Stir at 105 ° C for hours. The reaction was then cooled to room temperature and quenched by addition of a mixture of eastern skin 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 sulphate and filtered. The filtrate was concentrated in vacuo to give a yellow residue which was purified by column chromatography on silica gel eluting with 60 to 100% ethyl acetate in heptane. Mixture of the title compound and starting 5- (6-chloro-5-methylpyrimidin-4-yl) -1-methyl-1,4,5,6-tetrahydropyrrolo [3,4-c] pyrazole ( 1.20 g) was isolated as a white solid and used for further reaction without further purification.

Crude tert-butyl (3,4-cis) -3-fluoro-4-{[5-methyl-6- (1-methyl-4,6-dihydropyrrolo [] from a separate reaction carried out under the same conditions. The 3,4-c] pyrazol-5 (1H) -yl) pyrimidin-4-yl] oxy} piperidine-1-carboxylate batch was purified by HPLC. The crude sample (9.5 mg) was dissolved in dimethyl sulfoxide (1 mL) and 8.5 minutes with a linear gradient of 0% water / acetonitrile in 80% water / acetonitrile (0.03% ammonium hydroxide modifier) followed by 0% water / Waters XBridge C ₁₈ eluting with acetonitrile for 1.5 min (flow rate: 25 mL / min) Purification by preparative reverse phase HPLC on a 19 x 100 mm, 0.005 mm column. This gave the title compound (5 mg). Analytical LCMS: Retention time 2.81 min (Waters Xbridge C ₁₈ 4.6 x 50 mm, 0.005 mm column; 5% water / acetonitrile in 90% water / acetonitrile linear gradient over 4.0 minutes, then 5% water for 1 minute duration) Acetonitrile 0.03% ammonium hydroxide modifier; flow rate: 2.0 mL / min);

C. 5- (6-{[(3,4-cis) -3-fluoropiperidin-4-yl] oxy} -5-methylpyrimidin-4-yl) -1-methyl-1,4 Preparation of 5,6-tetrahydropyrrolo [3,4-c] pyrazole (racemic)

Crude tert-butyl (3,4-cis) -3-fluoro-4-{[5-methyl-6- (1-methyl-4,6-dihydropyrrolo [3,4-c] prepared above] Pyrazol-5 (1H) -yl) pyrimidin-4-yl] oxy} piperidine-1-carboxylate (1.20 g) is dissolved in dichloromethane (12 mL) and trifluoroacetic acid in this solution. (5 mL) was added. The reaction was stirred at room temperature for 1 hour. The solvent was removed in vacuo 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 1 N aqueous sodium hydroxide solution (20 mL) and extracted three times with dichloromethane (40 mL). The combined organic extracts were washed with brine, dried over sodium sulphate and filtered. The filtrate was concentrated under reduced pressure to afford 5- (6-{[(3,4-cis) -3-fluoropiperidin-4-yl] oxy} -5-methylpyrimidin-4-yl) as a white solid. -1-Methyl-1,4,5,6-tetrahydropyrrolo [3,4-c] pyrazole (0.72 g, 60% over 2 steps) was obtained and used without further purification.

Note that the example number starts at 11.

Example 11: Isopropyl 4- {5-cyano-4-[(2,4-difluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate

Isopropyl 4- (5-cyano-4-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) 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) are placed in a microwave vial, dissolved in acetonitrile (3 mL) and 20 min at 110 ° C. Heated in a microwave vial. The mixture was cooled to rt, concentrated in vacuo, 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 in vacuo. The crude product was purified by preparative reverse phase HPLC on a Waters Atlantis C ₁₈ column 4.6 x 50 mm, 0.005 mm eluting with a gradient of water in acetonitrile (0.05% trifluoroacetic acid modifier) and isopropyl 4- { 5-cyano-4-[(2,4-difluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate was obtained. Analytical LCMS: Retention time: 3.62 min (Waters Atlantis C ₁₈ 4.6 x 50 mm, 0.005 mm; 5% water / acetonitrile in 95% water / acetonitrile linear gradient over 4.0 minutes; 0.05% trifluoroacetic acid modifier; flow rate 2.0 mL / min);

Example 12 Isopropyl 4- {5-cyano-4-[(2-methylphenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate

Ortho-cresol (21 mg, 0.19 mmol) and isopropyl 4- (5-cyano-4-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) pi in acetonitrile (1.6 mL) To a stirred solution of ferridine-1-carboxylate (Preparation 10) (60 mg, 0.16 mmol) was added cesium carbonate (106 mg, 0.32 mmol). The mixture was heated at reflux for 15 h. After cooling to room temperature, the crude material was concentrated to dryness in vacuo and the residue was taken up in water and extracted three times with ethyl acetate (20 mL each). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and the filtrate was concentrated to dryness in vacuo to give a tan residue (0.065 g, 100%). The crude sample was dissolved in dimethyl sulfoxide (1 mL) and after a linear gradient of 0% water / acetonitrile in 80% water / acetonitrile for 8.5 minutes, followed by a 1.5 minute period of 0% water / acetonitrile (0.05% trifluoro Purification by preparative reverse phase HPLC on a Waters Sunfire C ₁₈ 19 × 100 mm, 0.005 mm column eluting with acetic acid modifier). Analytical LCMS: Retention time 3.82 minutes (Waters Atlantis C ₁₈ 4.6 x 50 mm, 0.005 mm column; 5% water / acetonitrile in 95% water / acetonitrile linear gradient over 4.0 minutes, then 5% water / aceto for 1 minute duration Nitrile, 0.05% trifluoroacetic acid modifier; flow rate: 2.0 mL / min);

Example 13: 1-methylcyclopropyl 4- {5-cyano-4-[(2,5-difluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-car Carboxylate

A) tert-butyl 4- (5-cyano-4-((2,5-difluorophenoxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

2,5-difluorophenol (54 mg, 0.39 mmol) and tert-butyl 4- (5-cyano-4-((methylsulfonyloxy) methyl) -1H-pyrazole-1 in 3 mL of acetonitrile To a stirred solution of -yl) piperidine-1-carboxylate (Preparation 16) (126 mg, 0.33 mmol) was added cesium carbonate (214 mg, 0.66 mmol). The mixture was heated at reflux for 15 h. The mixture was cooled to rt and diluted with ethyl acetate and water. The layers were separated and the aqueous phase 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-.ano-4-((2,5-difluorophenoxy) Methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate was obtained and used in the next step without purification.

B) 4-((2,5-difluorophenoxy) methyl) -1- (piperidin-4-yl) -1H-pyrazole-5-carbonitrile

Tert-butyl 4- (5-cyano-4-((2,5-difluorophenoxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxyl in 5 mL of dichloromethane To a solution of rate (137 mg, 0.33 mmol) was added 0.82 mL of hydrochloric acid (4M in 1,4-dioxane). After the mixture was stirred at room temperature for 2 hours, the mixture was concentrated in vacuo to afford 4-((2,5-difluorophenoxy) methyl) -1- (piperidin-4-yl) -1H-pyrazole -5-carbonitrile was obtained, which was used in the next step without purification.

C) 1-methylcyclopropyl 4- {5-cyano-4-[(2,5-difluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate

4-((2,5-difluorophenoxy) methyl) -1- (piperidin-4-yl) -1H-pyrazole-5-carbonitrile (104 mg, 0.33 mmol) in 3.3 mL of dichloromethane To a stirred solution of was added triethylamine (0.18 mL, 1.3 mmol) followed by 1-methylcyclopropyl 4-nitrophenyl carbonate (see Preparation 26 and WO09105717) (171 mg, 0.72 mmol). The resulting pale yellow mixture was stirred under nitrogen atmosphere for 15 hours. The reaction mixture was diluted with dichloromethane and water. The layers were separated and the aqueous phase 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. A portion of this material (45 mg) was dissolved in dimethyl sulfoxide (0.9 mL) and purified on a Waters Xbridge C ₁₈ column 19 x 100 mm, 0.005 column, eluting with a gradient of water in acetonitrile (0.03% ammonium hydroxide modifier). Purification by reverse phase HPLC. Analytical LCMS: retention time 3.60 minutes (Atlantis C ₁₈ 4.6 x 50 mm, 5 μιη column; 5% water / acetonitrile at 95% water / acetonitrile linear gradient over 4 minutes; 0.05% trifluoroacetic acid modifier; flow rate 2.0 mL / min;

Example 14 1-methylcyclopropyl 4- {5-cyano-4-[(2,3-difluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-car Carboxylate

The title compound was prepared using a commercially available 2,3-diflurophenol following procedures similar to Example 13. The crude material (49 mg) was dissolved in dimethyl sulfoxide (0.9 mL) and purified reverse phase on a Waters Xbridge C ₁₈ column 19 x 100 mm, 0.005 column, eluting with a gradient of water in acetonitrile (0.03% ammonium hydroxide modifier). Purification by HPLC. Analytical LCMS: retention time 3.62 min (Atlantis C ₁₈ 4.6 x 50 mm, 5 μιη column; 5% water / acetonitrile at 95% water / acetonitrile linear gradient over 4 minutes; 0.05% trifluoroacetic acid modifier; flow rate 2.0 mL / min;

Example 15 1-Methylcyclopropyl 4- {4-[(4-carbamoyl-2-fluorophenoxy) methyl] -5-cyano-1H-pyrazol-1-yl} piperidine- 1-carboxylate

A) tert-butyl 4- (4-((4-carbamoyl-2-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine-1-carboxyl Rate

Tert-butyl 4- (5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate in 3 mL of 1,4-dioxane (Preparation 15 ) In a stirred solution of (200 mg, 0.65 mmol), 3-fluoro-4-hydroxybenzamide (Preparation 23) (100 mg, 0.64 mmol) and triinilphosphine (188 mg, 0.72 mmol) Ethyl azodicarboxylate (0.11 mL, 0.69 mmol) was added dropwise. The resulting mixture was stirred at rt overnight, then the mixture was concentrated in vacuo. The residue was purified by flash chromatography, eluting with a 30 to 70% ethyl acetate gradient in heptanes to give tert-butyl 4- (4-((4-carbamoyl-2-fluorophenoxy) methyl as a white solid. ) -5-cyano-1H-pyrazol-1-yl) piperidine-1-carboxylate (215 mg) was obtained.

B) 4-((5-cyano-1- (piperidin-4-yl) -1H-pyrazol-4-yl) methoxy) -3-fluorobenzamide

Tert-butyl 4- (4-((4-carbamoyl-2-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine-1 in 2 mL dichloromethane To a stirred solution of carboxylate (215 mg, 0.48 mmol) 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% methanol in dichloromethane containing 2% aqueous ammonia to afford 4-((5-cyano-1- (piperidine) as a white solid. -4-yl) -1H-pyrazol-4-yl) methoxy) -3-fluorobenzamide (150 mg) was obtained.

C) 1-methylcyclopropyl 4- {4-[(4-carbamoyl-2-fluorophenoxy) methyl] -5-cyano-1H-pyrazol-1-yl} piperidine-1- Carboxylate

4-((5-cyano-1- (piperidin-4-yl) -1H-pyrazol-4-yl) methoxy) -3-fluorobenzamide (40 mg, 0.12 in 1 mL of dichloromethane) triethylamine (0.036 mL, 0.26 mmol) was added to the stirred solution of mmol), and then 1-methylcyclopropyl 4-nitrophenyl carbonate (Preparation 26 and WO09105717) (60 mg, 0.26 mmol) was added to room temperature. Was added. The resulting pale yellow mixture was stirred at 65 ° C. for 2 hours under a nitrogen atmosphere. The reaction was cooled to rt, 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 heptanes to give 1-methylcyclopropyl 4- {4-[(4-carbamoyl-2-fluorophenoxy as a white solid. ) Methyl] -5-cyano-1H-pyrazol-1-yl} piperidine-1-carboxylate (34 mg) was obtained.

1H NMR is present in less than 10% of what is considered to be �, the corresponding isopropyl carbamate derivative (from isopropyl 4-nitrophenyl carbonate contaminated with 1-methylcyclopropyl 4-nitrophenyl carbonate) Indicates.

Example 16: 1-methylcyclopropyl 4- {4-[(4-carbamoylphenoxy) methyl] -5-cyano-1H-pyrazol-1-yl} piperidine-1-carboxylate

The title compound was prepared using commercially available 4-hydroxybenzamide following a similar procedure as in Example 15.

1H NMR is present in less than 10% of what is considered to be �, the corresponding isopropyl carbamate derivative (from isopropyl 4-nitrophenyl carbonate contaminated with 1-methylcyclopropyl 4-nitrophenyl carbonate) Indicates.

Example 17 1-methylcyclopropyl 4- (5-cyano-4-((4-cyanophenoxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

The title compound was prepared using commercially available 4-hydroxybenzonitrile following a procedure similar to Example 15. Purification of the crude reaction mixture was carried out by flash chromatography eluting with a gradient mixture of ethyl acetate in heptane (0 to 100% ethyl acetate).

Example 18: Isopropyl 4- (4-((4- (1H-pyrazol-1-yl) phenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine-1 Carboxylate

The title compound was prepared using 4- (1H-pyrazol-1-yl) phenol (WO 2003072547) following a similar procedure as in Example 12. Purification of the crude reaction mixture was carried out by flash chromatography eluting with a gradient mixture of ethyl acetate in heptane (0 to 100% ethyl acetate).

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-pyra Zol-1-yl) piperidine-1-carboxylate

A) 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- (trimethylsilyl) ethoxy) methyl) -2H-tetrazol-5-yl) phenoxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

Isopropyl 4- (5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (94 mg, 0.322 in 5 mL of 1,4-dioxane) mmol), 2-fluoro-4- (1-((2- (trimethylsilyl) ethoxy) methyl) -1H-tetrazol-5-yl) phenol and 2-fluoro-4- (2-(( Stirred solution of 2- (trimethylsilyl) ethoxy) methyl) -2H-tetrazol-5-yl) phenol (Preparation 17) (100 mg, 0.322 mmol) and triphenylphosphine (110 mg, 0.42 mmol) To diethyl azodicarboxylate (0.060 mL, 0.39 mmol) was added dropwise. The resulting mixture was stirred at rt overnight, then the mixture was concentrated in vacuo. The residue was purified by flash chromatography, eluting with a gradient of 10 to 40% ethyl acetate in heptanes 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- (trimethylsilyl) ethoxy) methyl) -2H-tetrazol-5-yl) phenoxy) methyl) -1H- Pyrazol-1-yl) piperidine-1-carboxylate (140 mg, yield 74%) was obtained.

Isopropyl 4- (5-cyano-4-((2-fluoro-4- (2-((2- (trimethylsilyl) ethoxy) methyl) -2H-tetrazol-5-yl) phenoxy) Methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate.

B) Isopropyl 4- (5-cyano-4-((2-fluoro-4- (1H-tetrazol-5-yl) phenoxy) methyl) -1H-pyrazol-1-yl) piperi Dean-1-carboxylate and isopropyl 4- (5-cyano-4-((2-fluoro-4- (2H-tetrazol-5-yl) phenoxy) methyl) -1H-pyrazole- 1-yl) piperidine-1-carboxylate

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- (Trimethylsilyl) ethoxy) methyl) -2H-tetrazol-5-yl) phenoxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (220 mg, 0.38 mmol) Was dissolved in ethanol (3 mL) and an aqueous solution of 2M hydrochloric acid (3 mL) was added dropwise. The resulting mixture was stirred at 50 ° C. for 4 hours, then cooled to room temperature and filtered. The resulting white solid was washed with ethyl acetate and heptanes (1/1 volume) and dried under reduced pressure to give the title compound (80 mg, yield 47%).

Example 20 Isopropyl 4- (5-cyano-4-((2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenoxy) methyl) -1H-pyrazole- 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-pyrazole- 1-yl) piperidine-1-carboxylate

Isopropyl 4- (5-cyano-4-((2-fluoro-4- (1H-tetrazol-5-yl) phenoxy) methyl) -1H-pyra in tetrahydrofuran (2 mL) at room temperature Zol-1-yl) piperidine-1-carboxylate and isopropyl 4- (5-cyano-4-((2-fluoro-4- (2H-tetrazol-5-yl) phenoxy) To a solution of methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (70 mg, 0.15 mmol) was added sodium hydride (14 mg, 0.31 mmol) in two portions, and the resulting mixture was Was stirred for 5 minutes. Iodomethane (0.03 mL, 0.46 mmol) was then added and the reaction mixture was stirred for an additional 16 hours at room temperature. The reaction was quenched by the addition of water and the mixture was diluted with ethyl acetate. The organic phase was separated and the aqueous phase 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, yield 14%) and isopropyl 4- (5-cyano-4-((2-fluoro-4- (2-methyl-2H-tetrazol-5-yl) phenoxy) methyl) -1H-pyrazol-1-yl) piperi Dean-1 -carboxylate (30 mg, yield 42%) was obtained.

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).

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).

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

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

Isopropyl 4- (5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate in 5 mL of 1,4-dioxane (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 ) And triethylphosphine (77 mg, 0.29 mmol) were added dropwise to diethyl azodicarboxylate (0.046 mL, 0.28 mmol). The resulting mixture was stirred at rt for 15 h, then the mixture was concentrated in vacuo. The residue was purified by flash chromatography, eluting with a gradient of 5 to 40% ethyl acetate in heptanes 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, yield 86%) Obtained.

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) is dissolved in methanol (2 mL) and 4M hydrochloric acid solution in 1,4-dioxane (1 mL) Was added dropwise. The resulting mixture was stirred at rt for 2 h and then concentrated under reduced pressure. The residue (160 mg) was divided and about 50 mg of crude material was purified by reverse phase HPLC to give the title compound (30 mg, 26%) (column: Waters Xbridge C18 19 × 100, 5 μm; mobile phase A: 0.03% (v / v) ammonium hydroxide in water; mobile phase B: 0.03% (v / v) ammonium hydroxide in acetonitrile; gradient: 0% water / 100% aceto in 85% water / 15% acetonitrile linear for 8.5 minutes Nitrile, held in 0% water / 100% acetonitrile for 10.0 min.Flow rate: 25 mL / min Detection: 215 nm.

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

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

Isopropyl 4- (5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (43 mg, 0.15 in 3 mL of 1,4-dioxane) mmol), 2-fluoro-4- (1- (2- (trimethylsilyloxy) ethyl) -1H-tetrazol-5-yl) phenol (Preparation 20) (50 mg, 0.15 mmol) and triphenylforce To a stirred solution of pin (43 mg, 0.16 mmol) was added diethyl azodicarboxylate (0.025 mL, 0.16 mmol) dropwise. The resulting mixture was stirred at rt overnight, then the mixture was concentrated in vacuo. The residue was purified by flash chromatography, eluting with a gradient of 30 to 70% ethyl acetate in heptanes 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, yield 55%) Obtained.

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) is dissolved in methanol (2 mL) and 4M hydrochloric acid solution in 1,4-dioxane (1 mL) Was added dropwise. The resulting mixture was stirred at rt for 2 h and then concentrated under reduced pressure. The residue (60 mg) was purified by reverse phase HPLC to give the title compound (20 mg, yield 49%) (column: Waters Xbridge C18 19 × 100, 5 μm; mobile phase A: 0.03% in water (v / v) ammonium hydroxide; mobile phase B: 0.03% in acetonitrile (v / v) ammonium hydroxide; gradient: 0% water / 100% acetonitrile in 80% water / 20% acetonitrile linear for 8.5 minutes, 0% for 10.0 minutes Maintained in water / 100% acetonitrile flow rate: 25 mL / min detection: 215 nm

Example 24 1-methylcyclopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenoxy] methyl} -1H- Pyrazol-1-yl) piperidine-1-carboxylate

The title compound was prepared using 2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenol (Preparation 21) following a procedure similar to Example 15.

1 H NMR is present in less than 10% of what is considered to be the corresponding isopropyl carbamate derivative (from isopropyl 4-nitrophenyl carbonate containing 1-methylcyclopropyl 4-nitrophenyl carbonate) Indicates.

Example 25 1-methylcyclopropyl 4- (5-cyano-4-{[4- (1-methyl-1H-tetrazol-5-yl) phenoxy] methyl} -1H-pyrazol-1- I) piperidine-1-carboxylate

The title compound was prepared using 4- (1-methyl-1H-tetrazol-5-yl) phenol (Preparation 22) following a procedure similar to Example 15.

1H NMR is present in less than 10% of what is considered to be �, the corresponding isopropyl carbamate derivative (from isopropyl 4-nitrophenyl carbonate contaminated with 1-methylcyclopropyl 4-nitrophenyl carbonate) Indicates.

Example 26 1-Methylcyclopropyl 4- (4-((4-carbamoyl-3-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine- 1-carboxylate

The title compound was prepared using 2-fluoro-4-hydroxybenzamide (Preparation 24) following a procedure similar to Example 13.

1H NMR is present in less than 10% of what is considered to be �, the corresponding isopropyl carbamate derivative (from isopropyl 4-nitrophenyl carbonate contaminated with 1-methylcyclopropyl 4-nitrophenyl carbonate) Indicates.

Example 27: Isopropyl 4- (5-cyano-4- {1- [2-fluoro-4- (methylsulfonyl) phenoxy] ethyl} -1 H-pyrazol-1-yl) piperidine -1-carboxylate

The title compound was purified using 2-fluoro-4- (methylsulfonyl) phenol and isopropyl 4- (5-cyano-4- (1-hydroxyethyl) -1H-pyrazole- according to a similar procedure as in Example 15. 1-yl) piperidine-1-carboxylate (Preparation 25). Samples were purified by reverse phase HPLC (Column: Waters Xbridge C18 19 × 100, 5 μm; Mobile Phase A: 0.03% (v / v) in ammonium hydroxide; Mobile Phase B: 0.03% (v / v) in acetonitrile) Ammonium hydroxide: Gradient: 0% water / 100% acetonitrile in 80% water / 20% acetonitrile linear for 8.5 minutes, hold in 0% water / 100% acetonitrile for 10.0 minutes Flow: 25 mL / min.

Example 28: Isopropyl 4- (5-cyano-4- {1-[(2-methylpyridin-3-yl) oxy] ethyl} -1 H-pyrazol-1-yl) piperidine-1- Carboxylate

The title compound was washed with 2-methylpyridin-3-ol and isopropyl 4- (5-cyano-4- (1-hydroxyethyl) -1H-pyrazol-1-yl) following a procedure similar to Example 15. Prepared using ferridine-1-carboxylate (Preparation 25). Samples were purified by reverse phase HPLC (Column: Waters Xbridge C18 19 × 100, 5 μm; Mobile Phase A: 0.03% (v / v) in ammonium hydroxide; Mobile Phase B: 0.03% (v / v) in acetonitrile) Ammonium hydroxide: Gradient: 0% water / 100% acetonitrile in 80% water / 20% acetonitrile linear for 8.5 minutes, hold in 0% water / 100% acetonitrile for 10.0 minutes Flow: 25 mL / min.

Example 29: Isopropyl 4- (5-cyano-4- {2- [2-fluoro-4- (methylsulfonyl) phenyl] propyl} -1 H-pyrazol-1-yl) piperidine- 1-carboxylate

A) Isopropyl 4- (5-cyano-4-vinyl-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 ° C. add n-butyllithium (0.360 mL, 0.89 mmol, 2.5 M in hexane). Added dropwise. The resulting yellow mixture was stirred at −78 ° C. for 30 minutes and then isopropyl 4- (5-cyano-4-formyl-1H-pyrazol-1-yl) piperi in tetrahydrofuran (2.5 mL). A solution of dean-1-carboxylate (Example 9, Step A) (171 mg, 0.59 mmol) was added. The cold bath was removed and the reaction mixture was stirred at rt for 3.75 h. 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 afford the title compound (116 mg, 68%) as a clear oil.

B) (E, Z) -Isopropyl 4- (5-cyano-4- (2- (2-fluoro-4- (methylsulfonyl) phenyl) prop-1-enyl) -1H-pyrazole -1-yl) piperidine-1-carboxylate

Isopropyl 4- (5-cyano-4-vinyl-1H-pyrazol-1-yl) piperidine-1-carboxylate (116 mg, 0.4 mmol) and 2- in anhydrous dichloromethane (2 mL) In a solution of fluoro-4- (methylsulfonyl) -1- (prop-1-en-2-yl) benzene (Preparation 29) (43 mg, 0.20 mmol), the second generation Hoveyda-Grubbs ( Hoveyda-Grubbs) catalyst (commercially available from Aldrich) (12.5 mg, 0.020 mmol) was added. The green solution was heated at 40 ° C. for 72 hours with periodic addition of dichloromethane. The material was concentrated under reduced pressure and the residue was purified by silica gel chromatography (10 to 100% ethyl acetate in heptane) to give the product as an impurity oil (8 mg, 8%). The material was used as is.

C) Isopropyl 4- (5-cyano-4- {2- [2-fluoro-4- (methylsulfonyl) phenyl] propyl} -1H-pyrazol-1-yl) piperidine-1- Carboxylate

(E, Z) -Isopropyl 4- (5-cyano-4- (2- (2-fluoro-4- (methylsulfonyl) -phenyl) prop-1-enyl in ethyl acetate (3 mL) A solution of) -1H-pyrazol-1-yl) piperidine-1-carboxylate (8 mg, 0.02 mmol) was set up using a 10% palladium on carbon cartridge at a flow rate of 1 mL / min. Hydrogenated on an H-cube ™ at ". The material was concentrated in vacuo and the residue (4 mg) was purified by reverse phase HPLC (column: Waters Xbridge C18 19 × 100, 5 μm; mobile phase A: 0.03% (v / v) ammonium hydroxide in water; mobile phase B: 0.03% (v / v) ammonium hydroxide in acetonitrile; gradient: 90% water / 10% acetonitrile in linear 80% water / 20% acetonitrile for 8.5 minutes, 0% water / 100% acetonitrile up to 10 minutes Flow rate: 25 mL / min) The title compound (1.9 mg, 23%) was obtained:

Example 30: 1-methylcyclopropyl 4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl) piperidine-1 Carboxylate

The title compound was prepared using 2-methylpyridin-3-ol following a similar procedure as in 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 give 77 mg of the title compound as a white solid.

Example 31 1-Methylcyclopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1 Carboxylate

A) tert-butyl 4- (5-cyano-4-((2,3,6-trifluorophenoxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate

tert-butyl 4- (5-cyano-4-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) 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 a microwave vial and dissolved in acetonitrile (3 mL). The vial was heated in a microwave reactor at 110 ° C. 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 gradient of 0 to 30% ethyl acetate in heptanes to give tert-butyl 4- (5-cyano-4-((2,3,6-trifluorophenoxy) as a clear oil. 36.2 mg of c) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate was obtained.

B) 1-methylcyclopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-car Carboxylate

1-methylcyclopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate Was prepared using a commercially available 2,3,6-trifluoro phenol following procedures similar to Example 13 (B and C). The crude material (17.1 mg) was purified by preparative reverse phase HPLC on Sepax 2-ethyl pyridine column 250 × 21.2 mm, 0.005 eluting with a gradient of ethanol in heptane. Analytical LCMS: Retention time 11.769 minutes (Phenomenex Luna (2) C ₁₈ 150 x 3.0 mm, 5 μιη column; 100% methanol in 95% water / methanol linear gradient over 12.5 minutes; 0.1% formic acid modifier) Flow rate 0.75 mL / min;

Example 32: Isopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxyl Rate

The title compound was prepared using a commercially available 2,3,6-trifluorophenol following a similar procedure as in Example 11. The crude material was purified by column chromatography eluting with a gradient of 0 to 25% ethyl acetate in heptane to give isopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) as a clear oil. C) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate.

Example 33: Isopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1 H-imidazol-2-yl) phenoxy] methyl} -1 H-pyrazole- 1-yl) piperidine-1-carboxylate

The title compound was prepared in the same manner as in Example 11 with 2-fluoro-4- (1-methyl-1H-imidazol-2-yl) phenol (Preparation 28) and isopropyl 4- (5-cyano-4 Prepared from-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (Preparation 10). The crude material was purified by preparative reverse phase HPLC on Sepax Silica 250 × 21.2 mm, 0.005 mm eluting with a gradient of ethanol in heptane. Analytical LCMS: retention time 8.598 min (Phenomenex Luna (2) C ₁₈ 150 x 3.0 mm, 5 μιη column; 100% methanol in 95% water / methanol linear gradient over 12.5 minutes; 0.1% formic acid modifier; flow rate 0.75 mL) /minute;

Example 34 isopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-imidazol-5-yl) phenoxy] methyl} -1H-pyrazole- 1-yl) piperidine-1-carboxylate

The title compound was prepared in the same manner as in Example 11 with 2-fluoro-4- (1-methyl-1H-imidazol-5-yl) phenol (Preparation 27) and isopropyl 4- (5-cyano-4 Prepared from-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (Preparation 10). The crude material was purified by preparative reverse phase HPLC on Sepax silica 250 × 21.2 mm, 0.005 eluting with a gradient of ethanol in heptane. Analytical LCMS: Retention time 8.797 min (Phenomenex Luna (2) C ₁₈ 150 x 3.0 mm, 5 μιη column; 100% methanol in 95% water / methanol linear gradient over 12.5 minutes; 0.1% formic acid modifier; flow rate 0.75 mL) /minute;

Example 35 Isopropyl 4- [5-cyano-4-({[2-methyl-6- (1H-1,2,4-triazol-1-yl) pyridin-3-yl] oxy} methyl ) -1H-pyrazol-1-yl] piperidine-1-carboxylate

The title compound was prepared using 2-methyl-6- (1H-1,2,4-triazol-1-yl) pyridin-3-ol following a similar procedure as in Example 12. Samples were reversed phase HPLC (column: Waters Xbridge C18 19 × 100, 5 μm; mobile phase A: 0.03% (v / v) ammonium hydroxide in water; mobile phase B: 0.03% (v / v) ammonium hydroxide in acetonitrile; gradient : Purified by 0% water / 100% acetonitrile in 80% water / 20% acetonitrile linear for 8.0 min in 0% water / 100% acetonitrile until 9.5 min. Flow rate: 25 mL / min).

Example 36 Isopropyl 4- [5-cyano-4-({[2-methyl-6- (1H-1,2,4-triazol-1-yl) pyridin-3-yl] amino} methyl ) -1H-pyrazol-1-yl] piperidine-1-carboxylate

Isopropyl 4- (5-cyano-4-((methylsulfonyloxy) methyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate in 0.75 mL of tetrahydrofuran (Preparation 10 ) (44 mg, 0.12 mmol) was added N, N-diisopropylethylamine (0.042 mL, 0.24 mmol), followed by 2-methyl-6- (1H-1,2,4-tria). Zol-1-yl) pyridin-3-amine (21 mg, 0.12 mmol) was added. After the reaction mixture was heated at 60 ° C. for 16 hours, it was cooled to room temperature and diluted with water and brine. The mixture was then extracted three times with 15 mL of 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 yellow foam. Samples were purified by reverse phase HPLC (Column: Waters Sunfire C18 19 × 100, 5 μm; Mobile Phase A: 0.05% (v / v) trifluoroacetic acid in water; Mobile Phase B: 0.05% in acetonitrile (v / v) Trifluoroacetic acid; Gradient: 0% water / 100% acetonitrile in 90% water / 10% acetonitrile linear for 8.5 min, hold in 0% water / 100% acetonitrile until 10.0 min.Flow rate: 25 mL /minute.

Example 37: Isopropyl 4- [5-cyano-4-({[2-methyl-6- (methylsulfonyl) pyridin-3-yl] amino} methyl) -1H-pyrazol-1-yl] Piperidine-1-carboxylate

The title compound was prepared using 2-methyl-6- (methylsulfonyl) pyridin-3-amine following a similar procedure as in Example 36. Samples were purified by reverse phase HPLC (Column: Waters Xbridge C18 19 × 100, 5 μm; Mobile Phase A: 0.03% (v / v) in ammonium hydroxide; Mobile Phase B: 0.03% (v / v) in acetonitrile) Ammonium hydroxide; Gradient: 0% water / 100% acetonitrile in 85% water / 15% acetonitrile linear for 8.5 minutes, hold in 0% water / 100% acetonitrile up to 10.0 minutes Flow: 25 mL / min.

Example 38: 1-methylcyclopropyl 4- (5-cyano-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperi Dean-1-carboxylate

The title compound was prepared using commercially available 4-tetrazol-1-yl-phenol following a similar procedure as in Example 15. The crude material was purified by flash chromatography eluting with a gradient of 0% to 75% ethyl acetate in heptane.

Example 39: 1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazole-5-carbonitrile

A) tert-butyl 4- (5-cyano-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine-1- Carboxylate

To a stirred cold (0 ° C.) solution of triphenylphosphine (283 mg, 1.08 mmol) in tetrahydrofuran (2 mL) was added dropwise diethylazodicarboxylate (0.17 mL, 1.1 mmol). The cold reaction mixture was stirred for 20 minutes and then a solution of 4-tetrazol-1-yl-phenol (165.5 mg, 1.021 mmol) in tetrahydrofuran was added. After 35 minutes, tert-butyl 4- (5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl) piperidine-1-carboxylate in tetrahydrofuran (Preparation 15) (300 mg, 0.979 mmol) was added and the reaction was allowed to slowly warm to room temperature overnight. The reaction was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with a gradient of 0% to 80% ethyl acetate in heptane to afford the title compound (304 mg, 68%) as a white downy solid.

B) 1-piperidin-4-yl-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazole-5-carbonitrile

tert-butyl 4- (5-cyano-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine-1-carboxyl Rate (298 mg, 0.663 mmol) was dissolved in dichloromethane (1.6 mL). Trifluoroacetic acid (0.15 mL) was added and the reaction stirred for 1.5 h at room temperature under nitrogen. The reaction was concentrated and used as such in the next step without further purification.

C) 1-methylcyclopropyl 4- (5-cyano-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine- 1-carboxylate

1-piperidin-4-yl-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-5-carbonitrile (30 mg, 0.086 mmol) and di Isopropylethylamine (0.12 ml, 0.688 mmol) was dissolved in acetonitrile (2 mL) in a sealed tube. 2-chloro-5-ethylpyrimidine (0.020 mL, 0.2 mmol) was added and the reaction was heated at 120 ° C. for 18 hours and at room temperature for 36 hours. The reaction mixture was concentrated under reduced pressure and the crude was purified by flash chromatography eluting with a gradient of 0% to 70% ethyl acetate in heptane to give a brown solid. The solid was triturated with a minimum amount of ether to give the title compound (3 mg, 8%) as a light brown solid.

Example 40 Isopropyl 4- {5-cyano-4-[(3-cyanophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate

The title compound was prepared using a commercially available 3-cyanophenol following a similar procedure as in 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.

Example 41: Isopropyl 4- {5-cyano-4-[(4-cyano-3-methylphenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate

The title compound was prepared using 4-hydroxy-2-methylbenzonitrile following a procedure similar to Example 12. The crude was purified by flash chromatography eluting with a gradient of 0% to 40% ethyl acetate in heptanes to give 18.8 mg (69%) of the title compound as a clear residue.

Example 42: Isopropyl 4- {5-cyano-4-[(4-cyanophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate

The title compound was prepared using commercially available 4-cyanophenol following a similar procedure as in Example 12. The crude material was purified by flash chromatography eluting with a gradient of 0% to 40% ethyl acetate in heptanes to give 14.7 mg (56%) of the title compound as a sticky white solid.

Example 43: 4-[(4-cyano-2-fluorophenoxy) methyl] -1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -1H- Pyrazole-5-carbonitrile

The title compound was prepared using commercially available 4-cyano-2-fluorophenol following a similar procedure as in Example 39. The crude material was purified by flash chromatography eluting with a gradient of 0% to 1.5% methanol in dichloromethane. The resulting solid was further purified via recrystallization from 10% methanol / ethyl acetate to give 3.67 g (60%) of clear product as an almost white solid.

Example 44 tert-butyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-car Carboxylate

The title compound was prepared using commercially available 4-cyano-2-fluorophenol following a similar procedure as in Example 15. The crude material was purified by flash chromatography eluting with a gradient of 10% to 40% ethyl acetate in heptanes to afford the title compound (21 g, 100%).

Example 45: Isopropyl 4- {5-cyano-4-[(2-cyano-4-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxyl Rate

The title compound was prepared using a commercially available 2-cyano-4-fluorophenol following a similar procedure as in Example 15. The crude material was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 μιη; Modifier: 0.05% trifluoroacetic acid; Gradient: 95% water / 5% acetonitrile linear over 4.0 minutes 5% water / 95% acetonitrile, 5.0 Purification by minute holding in 5% water / 95% acetonitrile; flow rate: 2.0 mL / min) afforded 35.8 mg (73%) of the title compound.

Example 46: Isopropyl 4- (5-cyano-4-{[4- (dimethylcarbamoyl) -2-fluorophenoxy] methyl} -1 H-pyrazol-1-yl) piperidine- 1-carboxylate

The title compound was prepared using 3-fluoro-4-hydroxy-N, N-dimethylbenzamide (Preparation 31B) following a procedure similar to Example 15. The crude material was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 μιη; Modifier: 0.05% trifluoroacetic acid; Gradient: 95% water / 5% acetonitrile linear over 4.0 minutes 5% water / 95% acetonitrile, 5.0 Purification by minute holding in 5% water / 95% acetonitrile; flow rate: 2.0 mL / min) afforded 6.8 mg (12%) of the title compound.

Example 47 1-methylcyclopropyl 4- (5-cyano-4-{[4- (dimethylcarbamoyl) -2-fluorophenoxy] methyl} -1 H-pyrazol-1-yl) py Ferridine-1-carboxylate

The title compound was prepared using 3-fluoro-4-hydroxy-N, N-dimethylbenzamide (Preparation 31B) following a procedure similar to Example 15. The crude material was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 μιη; Modifier: 0.05% trifluoroacetic acid; Gradient: 5% water / 95% acetonitrile in 95% water / 5% acetonitrile linear over 4.0 minutes, Purification by 5 min water / 95% acetonitrile up to 5.0 min; flow rate: 2.0 mL / min) afforded 28.7 mg (51%) of the title compound.

Example 48 1-methylcyclopropyl 4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazol-1-yl) py Ferridine-1-carboxylate

The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide (Preparation 31A) following a procedure similar to Example 15. The crude material was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 μιη; Modifier: 0.05% trifluoroacetic acid; Gradient: 5% water / 95% acetonitrile in 95% water / 5% acetonitrile linear over 4.0 minutes, Purification by 5% water / 95% acetonitrile up to 5.0 min; flow rate: 2.0 mL / min) afforded 35.6 mg (65%) of the title product.

Example 49: 4-({5-cyano-1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -1H-pyrazol-4-yl} methoxy) -3-fluoro-N, N-dimethylbenzamide

The title compound was prepared using 3-fluoro-4-hydroxy-N, N-dimethylbenzamide (Preparation 31B) following a procedure similar to Example 39. The crude material was purified by HPLC (Column Waters Atlantis dC18 4.6x50 mm, 5 μιη; Modifier: 0.05% trifluoroacetic acid; Gradient: 5% water / 95% acetonitrile in 95% water / 5% acetonitrile linear over 4.0 minutes, Purification by 5% water / 95% acetonitrile up to 5.0 min; flow rate: 2.0 mL / min) gave 26.7 mg of the title product.

Example 50: 1-Methylcyclopropyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1 Carboxylate

The synthesis is outlined in Scheme 5 below.

Scheme 5

A) tert-butyl 4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

Ethyl 5-cyano-1H-pyrazole-4-carboxylate (Jubilant Chemsys Ltd., Noida 201 301 sector-59 di-12, Uttar Pradesh, 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) , Cooled to 0 ° C. A 40% solution of diethyl azodicarboxylate in toluene (76.5 mL, 420 mmol) was added dropwise When the addition was complete, the reaction was allowed to warm at room temperature over 1 hour and then 18 hours. Under vigorous stirring, heptane (1400 mL) was added carefully, and after 1 h a suspension formed, the solid was filtered off and the filtrate was heptane (400 mL) and ethyl acetate (200 mL). The mixture was washed with, then the filtrate was concentrated and the residue was taken up in 25% in heptane. Eluting with acetate and purified by flash chromatography, followed by ethyl acetate-to afford the desired product was recrystallized from heptane (35.2 g, 33%).

B): tert-butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate

tert-butyl 4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate (45.5 g, 131 mmol) was added to tetrahydrofuran ( 350 mL) and cooled to -78 ° C. A 1.5 M solution (50 g, 350 mmol) of diisobutylaluminum hydride in toluene was added dropwise over 75 minutes while maintaining the internal temperature between -65 ° C and -60 ° C. Once the addition was complete, the reaction mixture was warmed at -10 ° C for 90 minutes. While maintaining a temperature of −10 ° C., 4 M aqueous solution of potassium hydroxide (350 mL, 10.7 equiv) was added dropwise carefully. Once the addition was complete, the reaction mixture was allowed to warm slowly to room temperature with vigorous stirring and then allowed to stir at room temperature for 20 hours. Methyl tert-butyl ether (200 mL) and heptane (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 solid was filtered 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 ° C. for 1 hour and the suspension was slowly cooled to room temperature with stirring. The resulting solid was filtered and dried in a vacuum oven set at 40 ° C. to tert-butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1- Carboxylate (31.2 g, 78%) was obtained.

C) tert-butyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate

Tert-butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate (336 g, 1.10 mol) in a 4 L bottle, tri Phenylphosphine (359.58 g, 1.37 mol), 4-cyano-2-fluorophenol (157.89 g, 1.15 mol) and 2-methyltetrahydrofuran (2.02 L, 20.10 mol) were charged. The mixture was stirred into solution and kept under nitrogen at room temperature. Another 4 L bottle was charged with a solution of diethyl diazendicarboxylate in toluene (564.33 mL, 620.76 g, 1.43 mol) and 2-methyltetrahydrofuran (2.12 L, 21.11 mol). The mixture was stirred to complete solution and kept under nitrogen at room temperature. Pumping two streams into T-pieces (stainless) using a single peristaltic pump (two supply lines), then pumping into a coil volume of 100 mL (1/8 "followed by 1/4" ID PTE tube) Combined at a flow rate of 20 mL / min. After 8 hours of flow, the feed bottle was emptied and the bottle was rinsed using 2 x 25 mL of methyltetrahydrofuran and pumped through the line. The product stream was used as such for the next reaction.

D) 4-[(4-cyano-2-fluorophenoxy) methyl] -1-piperidin-4-yl-1H-pyrazole-5-carbonitrile tosylate salt

Tert-butyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1- collected in step C The stream of carboxylates was divided into two 5 L one-necked flasks. p-toluenesulfonic acid monohydrate (344.22 g, 1.81 mol) was charged in portions to the mixture and the flask was heated using a rotary evaporator bath maintained at 75 ° C. for 8 hours. The reaction was allowed to cool to room temperature and granulated overnight. The mixture was filtered and collected dry under vacuum for 3 hours to give the target product (480 g, 88% over 2 steps) as tosylate salt.

E) 1-methylcyclopropyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-car Carboxylate

Tosylate salt of 4-[(4-cyano-2-fluorophenoxy) methyl] -1-piperidin-4-yl-1H-pyrazole-5-carbonitrile (478 g, 960.71 mmol) Dissolved in 4-L bottle in 2-methyltetrahydrofuran (2.39 L, 23.83 mol) and water (478.00 mL). Triethylamine (200.86 mL, 1.44 mol) and 1-methylcyclopropyl 4-nitrophenyl carbonate (Preparation 26) (229.83 g, 960.71 mmol) were added and stirred for 48 hours. The reaction mixture was washed with aqueous 1 N sodium hydroxide (1 L). The mixture was stirred and the layers separated. The organic layer was washed several times with aqueous 1 N sodium hydroxide (1 L), dried over magnesium sulfate, filtered and the filtrate was concentrated under reduced pressure to give a light yellow solid. These solids were made into a slurry in ethyl acetate overnight at room temperature. The solid was filtered off and the resulting pale yellow solid was reslurried in ethyl acetate (3 vol), filtered and collected dry under vacuum to give the target compound (313 g, 77% in two batches) as an off-white solid. It was.

Melting point = 144.6 ° C

Combustion Analysis: Quantitative Technologies Inc. (QTI)

White House, New Jersey

Salem Ind. Park-Building 5

East 22 route 291

C22H22FN5O3

C (theoretical value = 62.40%)

62.28%

62.29%

H (theoretical value = 5.24%)

5.17%

5.13%

N (theoretical value = 16.54%)

16.42%

16.50%

Example 51 tert-butyl (3S, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazole-1 -Yl) -3-fluoropiperidine-1-carboxylate

A) tert-butyl (3S, 4S) -4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxyl Rate

tert-butyl (3S, 4S) -4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate For the preparation of tert-butyl 4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate (Example 50, Step A) Ethyl 5-cyano-1H-pyrazole-4-carboxylate and (3S, 4R) -tert-butyl 3-fluoro-4-hydroxypiperidine-1-carboxylate in a similar manner as described ( Prepared from Preparation Example 43 B). The crude material was purified by flash chromatography eluting with a gradient of 0% to 30% ethyl acetate in heptane to afford the desired product as a thick clear oil (149.4 mg, 32%).

B) tert-butyl (3S, 4S) -4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate

tert-butyl (3S, 4S) -4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-carboxylate -Butyl 4- [5-cyano-4- (hydroxymethyl) -1H-pyrazol-1-yl] piperidine-1-carboxylate (Example 50, Step B) as described for the preparation of Tert-butyl (3S, 4S) -4- [5-cyano-4- (ethoxycarbonyl) -1H-pyrazol-1-yl] -3-fluoropiperidine-1-car Prepared from carboxylates. The crude product was purified by flash chromatography eluting with a gradient of 5% to 50% ethyl acetate in heptane to afford the desired product as a thick clear oil (74 mg, 56%), which solidified upon standing.

C) tert-butyl (3S, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate

The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide (Preparation 31A) following a procedure similar to Example 50. Crude was purified by HPLC (Column: Waters Xbridge C12 4.6x50 mm, 5 μιη; Modifier: 0.05% ammonium hydroxide; Gradient: 95% water / 5% acetonitrile linear 4.0% 4.0 over 4.0 min, 5.0% acetonitrile, 5.0 Purification by 5% water / 95% acetonitrile up to minute; flow rate: 2.0 mL / min) afforded the desired product.

Example 52 52 tert-butyl (3R, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazole- 1-yl) -3-fluoropiperidine-1-carboxylate

The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide (Preparation 31A) following a procedure similar to Example 51. The crude material was purified by HPLC (column: Waters Xbridge C12 4.6x50 mm, 5 μιη; modifier: 0.05% ammonium hydroxide; gradient: 5% water / 95% acetonitrile in 95% water / 5% acetonitrile linear over 4.0 minutes, Purification by 5.0% holding in 5% water / 95% acetonitrile; flow rate: 2.0 mL / min) afforded the desired product.

Example 53: 1-Methylcyclopropyl (3S, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazole -1-yl) -3-fluoropiperidine-1-carboxylate

The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide (Preparation 31A) following procedures similar to Examples 50 and 51. Crude was purified by HPLC (Column: Princeton 2-ethyl pyridine 250 x 21.2 mm 5 μιη; Gradient: 95% heptane / 5% ethanol over 1.5 minutes, linear to 0% heptane / 100% ethanol over 10 minutes, 5.0 Purification via hold in 0% heptane / 100% ethanol (min) and linear to 95% heptane / 5% ethanol; flow rate 28 mL / min) until minutes afforded the desired product.

Example 54 1-methylcyclopropyl (3R, 4R) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazole -1-yl) -3-fluoropiperidine-1-carboxylate

The title compound was prepared using 3-fluoro-4-hydroxy-N-methylbenzamide (Preparation 31A) following procedures similar to Examples 50 and 51. The crude material was purified by HPLC (column: Princeton 2-ethyl pyridine 250 x 21.2 mm, 5 μιη; gradient: 95% heptane / 5% ethanol over 1.5 minutes, linear to 0% heptane / 100% ethanol over 10 minutes, up to 5.0 minutes Purification through 0% heptane / 100% ethanol (1 min) and 95% heptane / 5% ethanol linear; flow rate: 28 mL / min) afforded the desired product.

Example 55 tert-butyl (3S, 4S) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl)- 3-fluoropiperidine-1-carboxylate

tert-butyl 4- (5-cyano-4-{[(methylsulfonyl) oxy] methyl} -1H-pyrazol-1-yl) -3-fluoropiperidine-1-carboxylate (manufacture Example 42) (33 mg, 0.082 mmol) was dissolved in acetonitrile (3 mL) and cesium carbonate (53 mg, 0.164 mmol) and 3-hydroxy-2-methylpyridine (9 mg, 0.082 mmol) were added. . The reaction mixture was heated to 80 ° C. for 1.5 h. The reaction was cooled to rt 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 0.5 N sodium hydroxide, water and brine, 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 of 30% to 100% ethyl acetate in heptanes to give the racemic product (30 mg, 70%) as an amber oil.

Example 56 tert-butyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl)- 3-fluoropiperidine-1-carboxylate

The title compound was prepared using commercially available 3-hydroxy-2-methylpyridine following procedures similar to Example 55. The crude material was purified by flash chromatography eluting with a 30% to 100% ethyl acetate gradient in heptane to afford the desired product as an amber oil.

Example 57 1-methylcyclopropyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate

The title compound was prepared using commercially available 3-hydroxy-2-methylpyridine following procedures similar to Example 55. The crude material was purified by flash chromatography eluting with a gradient of 40% to 100% ethyl acetate in heptane to afford the desired racemic product as a white solid.

Example 58: 1-Methylcyclopropyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate

The title compound was prepared using commercially available 3-hydroxy-2-methylpyridine following procedures similar to Example 55. The crude material was purified by flash chromatography eluting with a gradient of 40% to 100% ethyl acetate in heptane to afford the racemic product, which was subjected to the following conditions: column: chiralcel OJ-H 4.6 mm x 25 cm; Mobile phase: 85/15 carbon dioxide / methanol, modifier: 0.2% isopropylamine; Flow rate: 2.5 mL / min further purification by chiral HPLC to give the title compound.

Example 59: 1-methylcyclopropyl (3R, 4S) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate

The title compound was prepared using commercially available 3-hydroxy-2-methylpyridine following procedures similar to Example 55. The crude material was purified by flash chromatography eluting with a gradient of 40% to 100% ethyl acetate in heptane to afford the racemic product, which was subjected to the following conditions: column: chiralcel OJ-H 4.6 mm x 25 cm; Mobile phase: 85/15 carbon dioxide / methanol, modifier: 0.2% isopropylamine; Flow rate: 2.5 mL / min further purification by chiral HPLC to give the title compound.

Example 60 tert-butyl 4- (5-cyano-4-{[4- (1H-1,2,3-triazol-1-yl) phenoxy] methyl} -1 H-pyrazole-1- I) piperidine-1-carboxylate

The title compound was purified using 4- (1H-1,2,3-triazol-1-yl) phenol (US Patent Application No. PCT / US2009 / 038315, Publication No. WO 2009/129036 A1), following procedures analogous to Example 15. Prepared using. Crude was purified by HPLC (column: Phenomenex Gemini C18 250x21.2 mm, 8 μιη; mobile phase: 50% acetonitrile (ammonia pH 10) in water (ammonia pH 10) in water (ammonia pH 10) Purification by 55% acetonitrile (ammonia pH 10); flow rate: 25 mL / min; wavelength: 220 nm) gave the title compound.

Example 61 tert-butyl 4- (5-cyano-4-{[4- (2H-1,2,3-triazol-2-yl) phenoxy] methyl} -1 H-pyrazole-1- I) piperidine-1-carboxylate

The title compound was purified using 4- (2H-1,2,3-triazol-2-yl) phenol (US Patent Application No. PCT / US2009 / 038315, Publication No. WO 2009/129036 A1), following procedures analogous to Example 15. Prepared using. Crude was purified by HPLC (column: Phenomenex Gemini C18 250x21.2 mm, 8 μιη; mobile phase: 63% acetonitrile (ammonia pH 10) in water (ammonia pH 10); flow rate: 25 mL / min; wavelength: 220 nm) Purification by gave the title compound.

Example 62 1-Methylcyclopropyl 4- (4-((4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -5-cyano-1H-pyrazole- 1-yl) piperidine-1-carboxylate

The title compound was prepared in a similar manner to Example 60 starting from Example 60. The crude material was purified by reverse phase HPLC:

Column: Chromasil Ethernity-5-C18 150x30 mmx 5 μm

Mobile phase: 58% acetonitrile (0.225% formic acid) in water (0.225% formic acid) in 38% acetonitrile (0.225% formic acid) in water (0.225% formic acid)

Flow rate: 30 mL / min

Wavelength: 220 nm

Example 63 1-Methylcyclopropyl 4- (4-((4- (2H-1,2,3-triazol-2-yl) phenoxy) methyl) -5-cyano-1H-pyrazole- 1-yl) piperidine-1-carboxylate

The title compound was prepared in a similar manner to Example 61 starting with Example 61. The crude residue was purified by preparative HPLC to give 50 mg (39%) of the title compound as a white solid:

Column: Boston Symmetrix ODS-H 150x30 mm x 5 μm

Mobile phase: 70% acetonitrile (0.225% formic acid) in water (0.225% formic acid) in 50% acetonitrile (0.225% formic acid) in water (0.225% formic acid)

Flow rate: 30 mL / min

Wavelength: 220 nm

Example 64 tert-butyl 4- [5-cyano-4-({[1- (methylsulfonyl) piperidin-4-yl] oxy} methyl) -1H-pyrazol-1-yl] pi Ferridine-1-carboxylate

The title compound was prepared in a similar manner to Example 13. The crude compound was purified by silica gel chromatography using a 1: 4 mixture of petroleum ether and ethyl acetate.

Example 65 tert-butyl 4- [5-cyano-4-({2-fluoro-4-[(2-hydroxyethyl) (methyl) carbamoyl] phenoxy} methyl) -1H-pyra Zol-1-yl] piperidine-1-carboxylate

The title compound was prepared in a similar manner to Example 46. The crude material was purified by reverse phase HPLC:

Column: Phenomenex Gemini C18 250x21.2 mm x 8 μm

Mobile phase: 60% acetonitrile (ammonia pH 10) in water (ammonia pH 10) in 40% acetonitrile (ammonia pH 10) in water (ammonia pH 10)

Flow rate: 25 mL / min

Wavelength: 220 nm

Example 66 tert-butyl 4- [5-cyano-4-({2-fluoro-4-[(3-hydroxypyrrolidin-1-yl) carbonyl] phenoxy} methyl) -1H -Pyrazol-1-yl] piperidine-1-carboxylate

The title compound was prepared in a similar manner to Example 46. The crude material was purified by reverse phase HPLC:

Column: Phenomenex Gemini C18 250x21.2 mm x 8 μm

Mobile phase: 60% acetonitrile (ammonia pH 10) in water (ammonia pH 10) in 40% acetonitrile (ammonia pH 10) in water (ammonia pH 10)

Flow rate: 25 mL / min

Wavelength: 220 nm

Example 67 tert-butyl 4- (4-{[4- (azetidin-1-ylcarbonyl) -2-fluorophenoxy] methyl} -5-cyano-1H-pyrazol-1-yl Piperidine-1-carboxylate

The title compound was prepared in a similar manner to Example 46. The crude material was purified by reverse phase HPLC:

Column: Phenomenex Gemini C18 250x21.2 mm x 8 μm

Mobile phase: 60% acetonitrile (ammonia pH 10) in water (ammonia pH 10) in 40% acetonitrile (ammonia pH 10) in water (ammonia pH 10)

Flow rate: 25 mL / min

Wavelength: 220 nm

Example 68 1-methylcyclopropyl 4- [5-cyano-4-({[1- (methylsulfonyl) piperidin-4-yl] oxy} methyl) -1H-pyrazol-1-yl Piperidine-1-carboxylate

The title compound was prepared in a similar manner to Example 64. The crude material was purified by reverse phase HPLC:

Column: Phenomenex Synergi C18 150x30 mm x 4 μm

Mobile phase: 53% acetonitrile (0.225% formic acid) in water (0.225% formic acid) in 43% acetonitrile (0.225% formic acid) in water (0.225% formic acid)

Flow rate: 30 mL / min

Various documents are cited throughout this application. The disclosure of which is incorporated herein by reference in its entirety 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.

Claims (12)

Isopropyl 4- {5-cyano-4-[(2,4-difluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;
Isopropyl 4- {5-cyano-4-[(2-methylphenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;
1-methylcyclopropyl 4- {5-cyano-4-[(2,5-difluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;
1-methylcyclopropyl 4- {5-cyano-4-[(2,3-difluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;
1-methylcyclopropyl 4- {4-[(4-carbamoyl-2-fluorophenoxy) methyl] -5-cyano-1H-pyrazol-1-yl} piperidine-1-carboxyl Rate;
1-methylcyclopropyl 4- {4-[(4-carbamoylphenoxy) methyl] -5-cyano-1H-pyrazol-1-yl} piperidine-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- I) 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-pyra Zol-1-yl) piperidine-1-carboxylate;
Isopropyl 4- (5-cyano-4-((2-fluoro-4- (1- (2-hydroxyethyl) -1H-tetrazol-5-yl) phenoxy) methyl) -1H-pyra Zol-1-yl) piperidine-1-carboxylate;
1-methylcyclopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenoxy] methyl} -1H-pyrazole-1 -Yl) piperidine-1-carboxylate;
1-methylcyclopropyl 4- (5-cyano-4-{[4- (1-methyl-1H-tetrazol-5-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperi Dine-1-carboxylate;
1-Methylcyclopropyl 4- (4-((4-carbamoyl-3-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine-1-carboxyl Rate;
Isopropyl 4- (5-cyano-4- {1- [2-fluoro-4- (methylsulfonyl) phenoxy] ethyl} -1H-pyrazol-1-yl) piperidine-1-car Carboxylates;
Isopropyl 4- (5-cyano-4- {1-[(2-methylpyridin-3-yl) oxy] ethyl} -1 H-pyrazol-1-yl) piperidine-1-carboxylate;
Isopropyl 4- (5-cyano-4- {2- [2-fluoro-4- (methylsulfonyl) phenyl] propyl} -1H-pyrazol-1-yl) piperidine-1-carboxyl Rate;
1-methylcyclopropyl 4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) piperidine-1-carboxylate ;
1-methylcyclopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate ;
Isopropyl 4- {5-cyano-4-[(2,3,6-trifluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;
Isopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-imidazol-2-yl) phenoxy] methyl} -1H-pyrazol-1-yl) Piperidine-1-carboxylate;
Isopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-imidazol-5-yl) phenoxy] methyl} -1H-pyrazol-1-yl) Piperidine-1-carboxylate;
Isopropyl 4- [5-cyano-4-({[2-methyl-6- (1H-1,2,4-triazol-1-yl) pyridin-3-yl] oxy} methyl) -1H- Pyrazol-1-yl] piperidine-1-carboxylate;
Isopropyl 4- [5-cyano-4-({[2-methyl-6- (1H-1,2,4-triazol-1-yl) pyridin-3-yl] amino} methyl) -1H- Pyrazol-1-yl] piperidine-1-carboxylate;
Isopropyl 4- [5-cyano-4-({[2-methyl-6- (methylsulfonyl) pyridin-3-yl] amino} methyl) -1H-pyrazol-1-yl] piperidine- 1-carboxylate;
1-methylcyclopropyl 4- (5-cyano-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine-1- Carboxylates;
1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyra Sol-5-carbonitrile;
Isopropyl 4- {5-cyano-4-[(3-cyanophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;
Isopropyl 4- {5-cyano-4-[(4-cyano-3-methylphenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate;
Isopropyl 4- {5-cyano-4-[(4-cyanophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate;
4-[(4-cyano-2-fluorophenoxy) methyl] -1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -1H-pyrazole-5 Carbonitrile;
tert-butyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate;
Isopropyl 4- {5-cyano-4-[(2-cyano-4-fluorophenoxy) methyl] -1 H-pyrazol-1-yl} piperidine-1-carboxylate;
Isopropyl 4- (5-cyano-4-{[4- (dimethylcarbamoyl) -2-fluorophenoxy] methyl} -1 H-pyrazol-1-yl) piperidine-1-carboxyl Rate;
1-methylcyclopropyl 4- (5-cyano-4-{[4- (dimethylcarbamoyl) -2-fluorophenoxy] methyl} -1 H-pyrazol-1-yl) piperidine-1 Carboxylates;
1-methylcyclopropyl 4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine-1 Carboxylates;
4-({5-cyano-1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -1H-pyrazol-4-yl} methoxy) -3-fluoro Rho-N, N-dimethylbenzamide;
1-methylcyclopropyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate ;
tert-butyl (3S, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazol-1-yl)- 3-fluoropiperidine-1-carboxylate;
tert-butyl (3R, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1 H-pyrazol-1-yl)- 3-fluoropiperidine-1-carboxylate;
1-methylcyclopropyl (3S, 4S) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate;
1-methylcyclopropyl (3R, 4R) -4- (5-cyano-4-{[2-fluoro-4- (methylcarbamoyl) phenoxy] methyl} -1H-pyrazol-1-yl ) -3-fluoropiperidine-1-carboxylate;
tert-butyl (3S, 4S) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3-fluoro Piperidine-1-carboxylate;
tert-butyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1 H-pyrazol-1-yl) -3-fluoro Piperidine-1-carboxylate;
1-methylcyclopropyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3- Fluoropiperidine-1-carboxylate;
1-methylcyclopropyl (3S, 4R) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3- Fluoropiperidine-1-carboxylate;
1-methylcyclopropyl (3R, 4S) -4- (5-cyano-4-{[(2-methylpyridin-3-yl) oxy] methyl} -1H-pyrazol-1-yl) -3- Fluoropiperidine-1-carboxylate;
tert-butyl 4- (5-cyano-4-{[4- (1H-1,2,3-triazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperi Dine-1-carboxylate;
tert-butyl 4- (5-cyano-4-{[4- (2H-1,2,3-triazol-2-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperi Dine-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 4- [5-cyano-4-({[1- (methylsulfonyl) piperidin-4-yl] oxy} methyl) -1H-pyrazol-1-yl] piperidine-1 Carboxylates;
tert-butyl 4- [5-cyano-4-({2-fluoro-4-[(2-hydroxyethyl) (methyl) carbamoyl] phenoxy} methyl) -1H-pyrazole-1- General] piperidine-1-carboxylate;
tert-butyl 4- [5-cyano-4-({2-fluoro-4-[(3-hydroxypyrrolidin-1-yl) carbonyl] phenoxy} methyl) -1H-pyrazole- 1-yl] piperidine-1-carboxylate;
tert-butyl 4- (4-{[4- (azetidin-1-ylcarbonyl) -2-fluorophenoxy] methyl} -5-cyano-1H-pyrazol-1-yl) piperidine -1-carboxylate;
1-methylcyclopropyl 4- [5-cyano-4-({[1- (methylsulfonyl) piperidin-4-yl] oxy} methyl) -1H-pyrazol-1-yl] piperidine -1-carboxylate;
1-methylcyclopropyl 4- (5-cyano-4-((2-fluoro-4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -1H-pyrazole -1-yl) piperidine-1-carboxylate;
Isopropyl 4- (5-cyano-4-((2-fluoro-4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -1H-pyrazole-1- I) piperidine-1-carboxylate;
1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-fluoro-4- (1H-1,2,3-triazole-1- Yl) phenoxy) methyl) -1H-pyrazole-5-carbonitrile;
Isopropyl 4- (4-((4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine -1-carboxylate;
4-((4- (1H-1,2,3-triazol-1-yl) phenoxy) methyl) -1- (1- (5-ethylpyrimidin-2-yl) piperidine-4- Yl) -1H-pyrazole-5-carbonitrile;
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-ethylpyrimidin-2-yl) piperidine-4- Yl) -1H-pyrazole-5-carbonitrile;
1-Methylcyclopropyl 4- (5-cyano-4-((2-fluoro-4- (2H-1,2,3-triazol-2-yl) phenoxy) methyl) -1H-pyrazole -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- I) piperidine-1-carboxylate;
1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-fluoro-4- (2H-1,2,3-triazole-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-pyrazole- 1-yl) piperidine-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) pi Ferridine-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-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((3-fluoro-4- (1H-tetrazol-1-yl) phenoxy) methyl ) -1H-pyrazole-5-carbonitrile;
4-((5- (1H-1,2,3-triazol-1-yl) pyridin-2-yloxy) methyl) -1- (1- (5-ethylpyrimidin-2-yl) piperi Din-4-yl) -1H-pyrazole-5-carbonitrile;
1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-methyl-6- (1H-1,2,3-triazol-1-yl ) Pyridin-3-yloxy) methyl) -1H-pyrazole-5-carbonitrile;
Isopropyl 4- (5-cyano-4-((2-methyl-6- (1H-1,2,3-triazol-1-yl) pyridin-3-yloxy) methyl) -1H-pyrazole -1-yl) piperidine-1-carboxylate;
1-methylcyclopropyl 4- (5-cyano-4-((2-methyl-6- (1H-1,2,3-triazol-1-yl) pyridin-3-yloxy) methyl) -1H -Pyrazol-1-yl) piperidine-1-carboxylate;
1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -4-((2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) Phenoxy) methyl) -1H-pyrazole-5-carbonitrile;
1-Methylcyclopropyl 4- (4-((4- (azetidine-1-carbonyl) -2-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperi Dine-1-carboxylate;
Isopropyl 4- (4-((4- (azetidine-1-carbonyl) -2-fluorophenoxy) methyl) -5-cyano-1H-pyrazol-1-yl) piperidine-1 Carboxylates; And
4-((4- (azetidine-1-carbonyl) -2-fluorophenoxy) methyl) -1- (1- (5-ethylpyrimidin-2-yl) piperidin-4-yl) -1H-pyrazole-5-carbonitrile
&Lt; / RTI &gt; or a pharmaceutically acceptable salt thereof. 1-methylcyclopropyl 4- (5-cyano-4-{[2-fluoro-4- (1-methyl-1H-tetrazol-5-yl) phenoxy] methyl} -1H-pyrazole-1 -Yl) piperidine-1-carboxylate;
1-methylcyclopropyl 4- (5-cyano-4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyrazol-1-yl) piperidine-1- Carboxylates;
1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -4-{[4- (1H-tetrazol-1-yl) phenoxy] methyl} -1H-pyra Sol-5-carbonitrile;
4-[(4-cyano-2-fluorophenoxy) methyl] -1- [1- (5-ethylpyrimidin-2-yl) piperidin-4-yl] -1H-pyrazole-5 Carbonitrile; And
1-methylcyclopropyl 4- {5-cyano-4-[(4-cyano-2-fluorophenoxy) methyl] -1H-pyrazol-1-yl} piperidine-1-carboxylate
&Lt; / RTI &gt; or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a compound according to claim 1 or 2 present in a therapeutically effective amount in admixture with one or more pharmaceutically acceptable excipients. The composition of claim 3, further comprising one or more additional pharmaceutical agents selected from the group consisting of anti-obesity agents and anti-diabetic agents. The anti-obesity agent according to claim 4, wherein the anti-obesity agent is dilottapid, mitratapid, implitapide, R56918 (CAS No. 403987), CAS No. 913541-47-6, lorcaserin, cetilistat, PYY _3-36 , Naltrexone, oleoyl-estrone, obinetidet, pramlinet, tesofensin, leptin, liraglutide, bromocriptine, orlistat, exenatide, AOD-9604 (CAS No. 221231-10-3) and The composition is selected from the group consisting of sibutramine. The method according to claim 4, wherein the antidiabetic agent is metformin, acetohexamide, chlorpropamide, diabines, glybenclamide, glyphide, glyburide, glymepiride, glyclazide, glypentide, glyquidone , Glysolamide, tolazamide, tolbutamide, tendamistat, trestatin, acarbose, adiposin, camigliose, emiglitate, miglitol, bogliboss, pradimycin-Q, salvostatin, Balaglitazone, Ciglitazone, Darglitazone, Englitazone, Isaglitazone, Pioglitazone, Rosiglitazone, Troglitazone, Exendin-3, Exendin-4, Trodusquemin, Reserbatrol, Hirtio A composition selected from the group consisting of flesh extract, cytagliptin, bilagliptin, allogliptin and saxagliptin. A method of treating diabetes, comprising administering an effective amount of a compound according to claim 1 to a patient in need thereof. A method of treating a metabolic or metabolic-related disease, condition or disorder comprising administering to a patient a therapeutically effective amount of a compound according to claim 1. Hyperlipidemia, type I diabetes, type II diabetes, idiopathic type I diabetes (type Ib), adult latent autoimmune diabetes, early, comprising administering an effective amount of a compound according to claim 1 or 2 Onset type 2 diabetes, adolescent-onset atypical diabetes, adolescent onset diabetes, malnutrition-related diabetes, gestational diabetes, coronary heart disease, ischemic stroke, restenosis after angioplasty, peripheral vascular disease, intermittent claudication, myocardium Infarction (e.g., necrosis and apoptosis), dyslipidemia, postprandial hemostasis, impaired glucose tolerance, fasting plasma glycemic disorder status, metabolic acidosis, ketoneosis, arthritis, obesity, osteoporosis, hypertension, congestive heart failure, left ventricular hypertrophy, peripheral arterial disease , Diabetic retinopathy, macular degeneration, cataracts, diabetic nephropathy, glomerulosclerosis, chronic renal failure, diabetic neuropathy, metabolic syndrome, syndrome X, premenstrual syndrome, coronary Pulsed heart disease, angina pectoris, thrombosis, atherosclerosis, myocardial infarction, transient ischemic attack, stroke, vascular restenosis, hyperglycemia, hyperinsulinemia, hyperlipidemia, hypertriglyceridemia, insulin resistance, impaired glucose metabolism, impaired glucose tolerance, fasting plasma Hypoglycemic conditions, obesity, erectile dysfunction, skin and connective tissue disorders, foot ulcers and ulcerative colitis, endothelial dysfunction and vascular elasticity, hyperapolipoproteinemia, Alzheimer's disease, schizophrenia, cognitive disorders, inflammatory bowel disease, A method of treatment in a condition selected from the group consisting of ulcerative colitis, Crohn's disease and irritable bowel syndrome. To patients in need of treatment of a metabolic or metabolic-related disease, condition or disorder
(i) a first composition according to claim 3; And
(ii) a second composition comprising one or more additional pharmaceutical agents selected from the group consisting of anti-obesity agents and anti-diabetic agents and one or more pharmaceutically acceptable excipients
A method of treating a metabolic or metabolic-related disease, condition or disorder comprising administering two separate pharmaceutical compositions comprising. The method of claim 10, wherein the first composition and the second composition are administered simultaneously. The method of claim 10, wherein the first composition and the second composition are administered sequentially and in any order.