Method and intermediate for preparing 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt

Abstract

The present invention provides a method for preparing 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt, certain intermediates, and processes for preparing these intermediates.

Description

Technical Field

[0001] The present invention provides a method for preparing 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt, certain intermediates, and processes for preparing these intermediates.

Background Art

[0002] Diabetes is a major public health concern due to its increasing prevalence and associated health risks. The disease is characterized by high levels of blood glucose resulting from defects in insulin production, insulin action, or both. Two major forms of diabetes, type 1 and type 2, are recognized. Type 1 diabetes (T1D) develops when the body's immune system destroys pancreatic beta cells, the only cells in the body that produce the hormone insulin that regulates blood glucose. People with type 1 diabetes must receive insulin by injection or pump to survive. Type 2 diabetes mellitus (generally referred to as T2DM) usually begins either with insulin resistance or insufficient insulin production to maintain acceptable glucose levels.

[0003] Currently, various pharmacological approaches are available for treating hyperglycemia and subsequent T2DM (Hampp, C. et al., Use of Antidiabetic Drugs in the U.S., 2003 - 2012, Diabetes Care 2014, 37, 1367 - 1374). These can be grouped into six major classes that act via different primary mechanisms: (A) Insulin secretagogues, including sulfonylureas (e.g., glypidide, glimepiride, glipizide), meglitinides (e.g., nateglinide, repaglinide), dipeptidyl peptidase IV (DPP-IV) inhibitors (e.g., sitagliptin, vildagliptin, alogliptin, dutogliptin, linagliptin, saxagliptin), and glucagon-like peptide-1 receptor (GLP-1R) agonists (e.g., liraglutide, albiglutide, exenatide, lixisenatide, dulaglutide, semaglutide), which act on pancreatic beta cells to enhance insulin secretion. Sulfonylureas and meglitinides have limited efficacy and tolerability, cause weight gain, and often induce hypoglycemia. DPP-IV inhibitors have limited efficacy. The marketed GLP-1R agonists are peptides administered by subcutaneous injection. Liraglutide is additionally approved for the treatment of obesity. (B) Biguanides (e.g., metformin) are thought to act primarily by decreasing hepatic glucose production. Biguanides often cause gastrointestinal disorders and lactic acidosis, further limiting their use. (C) Alpha-glucosidase inhibitors (e.g., acarbose) reduce intestinal glucose absorption. These agents often cause gastrointestinal disorders. (D) Thiazolidinediones (e.g., pioglitazone, rosiglitazone) act on specific receptors (peroxisome proliferator-activated receptor-gamma) in the liver, muscle, and adipose tissue. They regulate lipid metabolism and then enhance the response of these tissues to insulin action. Frequent use of these drugs may lead to weight gain and can induce edema and anemia.(E) Insulin is used, either alone or in combination with the above-mentioned agents, in more severe cases, and frequent use may also lead to weight gain and carries the risk of hypoglycemia. (F) Sodium-glucose co-transporter 2 (SGLT2) inhibitors (e.g., dapagliflozin, empagliflozin, canagliflozin, ertugliflozin) inhibit the reabsorption of glucose in the kidneys, thereby reducing glucose levels in the blood. This emerging class of drugs may be associated with ketoacidosis and urinary tract infections.

[0004] However, except for GLP-1R agonists and SGLT2 inhibitors, drugs have limited efficacy and do not address the most important problem of declining β-cell function and associated obesity.

[0005] Obesity is a highly prevalent chronic disease in modern society and is associated with a number of medical problems, including hypertension, hypercholesterolemia, and coronary heart disease. It is also highly correlated with T2DM and insulin resistance, the latter of which is generally accompanied by hyperinsulinemia or hyperglycemia, or both. In addition, T2DM is associated with a 2- to 4-fold increased risk of coronary artery disease. Currently, the only treatment that effectively eliminates obesity is bariatric surgery, which is expensive and risky. Pharmacological interventions are generally not very effective and are associated with side effects. Therefore, there is clearly a need for more effective pharmacological interventions with fewer side effects and convenient administration.

[0006] T2DM is most commonly associated with hyperglycemia and insulin resistance, but other diseases associated with T2DM include hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, obesity, dyslipidemia, hypertension, hyperinsulinemia, and non-alcoholic fatty liver disease (NAFLD).

[0007] NAFLD is a hepatic manifestation of metabolic syndrome and is a spectrum of liver conditions that includes steatosis, non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and ultimately hepatocellular carcinoma. NAFLD and NASH are considered primary fatty liver diseases because they account for the largest proportion of individuals with elevated liver lipids. The severity of NAFLD / NASH is based on the presence of lipids, inflammatory cell infiltration, hepatocyte ballooning, and the degree of fibrosis. Not all individuals with steatosis progress to NASH, but a significant proportion do.

[0008] GLP-1 is a 30-amino acid incretin hormone secreted by L cells in response to food intake in the gut. GLP-1 has been shown to stimulate insulin secretion in a physiological and glucose-dependent manner, decrease glucagon secretion, inhibit gastric emptying, reduce appetite, and stimulate beta cell proliferation. In preclinical experiments, GLP-1 promotes the persistence of beta cell function by stimulating the transcription of genes important for glucose-dependent insulin secretion and by promoting beta cell neogenesis (Meier et al., Biodrugs. 2003;17(2):93-102).

[0009] In healthy individuals, GLP-1 plays an important role in regulating postprandial blood glucose levels by stimulating glucose-dependent insulin secretion by the pancreas, resulting in increased glucose uptake peripherally. GLP-1 also suppresses glucagon secretion, leading to a reduction in hepatic glucose production. In addition, GLP-1 delays gastric emptying and slows small intestinal motility, delaying food absorption. In people with T2DM, the normal postprandial rise in GLP-1 is absent or reduced (Vilsboll T et al., Diabetes. 2001.50;609-613).

[0010] Holst (Physiol. Rev. 2007, 87, 1409) and Meier (Nat. Rev. Endocrinol. 2012, 8, 728) have described three major pharmacological activities of GLP-1 receptor agonists such as GLP-1, liraglutide, and exenatide-4 to improve glycemic control in patients with T2DM by reducing fasting and postprandial glucose (FPG and PPG): (i) increased glucose-dependent insulin secretion (improvement in the first and second phases), (ii) glucagon inhibitory activity under hyperglycemic conditions, and (iii) delayed gastric emptying rate resulting in delayed absorption of dietary glucose.

Summary of the Invention

Problems to be Solved by the Invention

[0011] Simple preventive and / or treatment measures for cardiometabolic and related diseases are still needed.

[0012] The compound 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid is a GLP-1R agonist described in U.S. Patent No. 10,208,019 (see Example 4A-01 of the patent), the disclosure of which is hereby incorporated by reference in its entirety for all purposes. The compound designated herein as "C111" has the following structure:

[0013]

Chemical formula

[0014] The compound may be administered in a pharmaceutically acceptable salt form thereof, for example, as its 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt (also known as its 2-amino-2-(hydroxymethyl)propane-1,3-diol salt), or as its tris(hydroxyethyl)methylamine salt, or as its tris salt.

[0015] The tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid means the salt of C111 prepared by using 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine. Tris relates to the carboxylic acid moiety of C111. Unless otherwise stated, when referring to the tris salt of C111, the counterion and C111 are in a stoichiometric ratio of about 1:1 (i.e., from 0.9:1.0 to 1.0:0.9, for example, from 0.95:1.00 to 1.00:0.95, or from 0.99:1.00 to 1.00:1.01). Another chemical name representing the tris salt of C111 is 1,3-dihydroxy-2-(hydroxymethyl)propan-2-aminium 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate, which may also be represented, for example, by one of the following structures.

[0016]

Chemical Structure

[0017] The solid forms of a particular drug, such as crystalline forms (including, for example, anhydrates, hydrates, solvates, etc.), are often well-known to be important determinants of the ease of drug preparation, stability, solubility, storage stability, ease of formulation, ease of handling, as well as in vivo pharmacology and / or efficacy. When a substance of the same composition crystallizes in different lattice arrangements, different crystalline forms appear, resulting in different thermodynamic properties and stabilities specific to the particular polymorphic forms. In cases where two or more solid forms (e.g., two or more crystalline forms, or an amorphous form and one or more crystalline forms) can be produced, it is desirable to have a method for producing each of the solid forms in a pure form. When determining which solid form is preferred, numerous properties of the solid forms must be compared, and the preferred solid (e.g., crystalline) form must be selected based on many physical property variables. It is entirely possible that one crystalline form may be preferred in some situations where certain aspects such as ease of preparation, stability, etc. are critical. In other situations, different crystalline forms may be preferred for greater solubility and / or better pharmacokinetics. Furthermore, due to the potential advantages associated with one pure crystalline form, when two or more solid forms of a substance can exist, it is desirable to prevent or minimize polymorphic transformation (i.e., the transformation from one crystalline form to another, or the transformation between one crystalline form and an amorphous form). Such polymorphic transformation can occur both during the preparation of a formulation containing a solid form (e.g., a crystalline form) and during the storage of a pharmaceutical dosage form containing a solid form (e.g., a crystalline form). For example, since improved drug formulations showing better bioavailability or better stability are consistently desired, there is a continuing need for new or more pure solid (e.g., crystalline) forms of existing drug molecules. Furthermore, there is a continuing need for cheaper and / or more efficient processes (including those that improve purity and / or generate fewer unwanted impurities) for making existing drug molecules. The processes and intermediates described herein are directed to this and other important objectives.

Means for Solving the Problems

[0018] In one embodiment (Embodiment A1), the present invention is a process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile,

[0019]

Chemical formula

[0020] In one embodiment (Embodiment B1), the present invention is a process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile,

[0021] [Chemical formula] (a1)tert-Butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate is reacted with 3-fluoro-4-(hydroxymethyl)benzonitrile, a palladium catalyst [palladium(II) acetate, tris(dibenzylideneacetone)dipalladium, or palladium(II) chloride, etc.], a base [an inorganic base, e.g., tripotassium phosphate, cesium carbonate, potassium hydroxide, or sodium hydride, etc.], and a phosphorus ligand [a monodentate phosphorus ligand or a bidentate phosphorus ligand, e.g., 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (XPhos), (2-biphenylyl)di-tert-butylphosphine (JohnPhos), 5-(di-tert-butylphosphino)-1′,3′,5′-triphenyl-1′H-[1,4′]bipyrazole (Bippyphos), 5-[bis(1-adamantyl)phosphino]-1′,3′,5′-triphenyl-1,4′-bi-1H-pyrazole (AdBippyphos), 2-(dicyclohexylphosphino)-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl (BrettPhos), di(adamantan-1-yl)(2′,4′,6′-triisopropyl-3,6-dimethoxy-2-biphenylyl)phosphine (AdBrettPhos), 2-dicyclohexylphosphino-2′,6-bis(N,N-dimethylamino)biphenyl (CPhos), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (SPhos), 2-dicyclohexylphosphino-2′-(N,N-dimethylamino)biphenyl (DavePhos), 2-di-tert-butylphosphino-2′-(N,N-dimethylamino)biphenyl (tBuDavePhos), 2-(di-tert-butylphosphino)-2′,4′,6′-triisopropyl-3,6-dimethoxy-1,1′-biphenyl (tBuBrettPhos), di-tert-butyl(2′,4′,6′-triisopropyl-3-methoxy-6-methyl-[1,1′-biphenyl]-2-yl)phosphine (RockPhos), 4,5-bis(diphenylphosphino)-9,In the presence of 9 - dimethylxanthene (XantPhos), or bis[(2 - diphenylphosphino)phenyl]ether (DPEPhos), etc., in a solvent system [such as those containing anisole, acetonitrile, tetrahydrofuran, dioxane, or methyl tert - butyl ether, etc.], react to form tert - butyl 4-(6 - ((4 - cyano - 2 - fluorobenzyl)oxy)pyridin - 2 - yl)piperidine - 1 - carboxylate, where the amount of 3 - fluoro - 4-(hydroxymethyl)benzonitrile is about 1.0 to about 1.1 molar equivalents relative to tert - butyl 4-(6 - chloropyridin - 2 - yl)piperidine - 1 - carboxylate, and, (a2) At the completion of the reaction in step (a1), add water, ethyl acetate, and ethanol to the reaction mixture, (a3) Separate the organic phase from the aqueous phase from step (a2), (b1) Add p - toluenesulfonic acid monohydrate to the separated organic phase from step (a3), thereby reacting tert - butyl 4-(6 - ((4 - cyano - 2 - fluorobenzyl)oxy)pyridin - 2 - yl)piperidine - 1 - carboxylate with p - toluenesulfonic acid monohydrate to form the bis(4 - methylbenzenesulfonate) salt of 3 - fluoro - 4 -(((6 - (piperidin - 4 - yl)pyridin - 2 - yl)oxy)methyl)benzonitrile, where the amount of p - toluenesulfonic acid monohydrate is about 2.0 to about 3.0 molar equivalents relative to tert - butyl 4-(6 - ((4 - cyano - 2 - fluorobenzyl)oxy)pyridin - 2 - yl)piperidine - 1 - carboxylate, (b2) Isolate the bis(4 - methylbenzenesulfonate) salt of 3 - fluoro - 4 -(((6 - (piperidin - 4 - yl)pyridin - 2 - yl)oxy)methyl)benzonitrile from step (b1), A process is provided that includes the above steps.

[0022] In one embodiment (Embodiment C1), the present invention is a process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile,

[0023]

Chemical formula

[0024] In one embodiment (Embodiment D1), the present invention provides anhydrous 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt, the monohydrate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt, the monotosylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, the bismesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The monomesylate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The monosulfate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The hemisulfate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, Crystalline methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, or (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid hemibarium salt which provides an intermediate useful for preparing C111 or a tris salt of C111.

[0025] In one embodiment (Embodiment E1), the present invention is a process for preparing methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, comprising:

[0026] [Chemical formula] (a1) preparing methyl (S)-4-amino-3-(oxetane-2-carboxamide)benzoate; (a2) reducing methyl (S)-4-amino-3-(oxetane-2-carboxamide)benzoate in the presence of a reducing reagent to form methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate; Step of reacting methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate with 2-chloro-1,1,1-trimethoxyethane in the presence of an acid (such as an organic acid like citric acid or p-toluenesulfonic acid) to form methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate A process is provided that includes

[0027] In one embodiment (Embodiment F1), the present invention is a process for preparing methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, comprising

[0028] [Chemical formula] (a1) Methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate is reacted with the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in a solvent system containing acetonitrile in the presence of diisopropylethylamine to form methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, where the amount of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is from about 1.1 to about 1.5 molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and the amount of diisopropylethylamine is from about 4.0 to about 6.0 (e.g., 5.0) molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. A process is provided that includes

[0029] In one embodiment (Embodiment G1), the present invention is a process for preparing methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, which comprises

[0030] [Chemical formula] (a1) Methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate is reacted with the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in a solvent system containing methanol in the presence of diisopropylethylamine to form methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, where the amount of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is about 1.0 to about 1.2 (e.g., 1.1) molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and the amount of diisopropylethylamine is about 4.0 to about 6.0 (e.g., 5.0) molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. A process is provided which includes this.

[0031] In one embodiment (Embodiment H1), the present invention is a process for preparing the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid,

[0032] [Chemical formula] (a1) Reacting methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with barium hydroxide [Ba(OH)₂] in a solvent system containing an organic solvent and water to form the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, wherein the amount of barium hydroxide [Ba(OH)₂] is about 0.5 to about 0.6 (e.g., about 0.5) molar equivalents relative to methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and (a2) Optionally, isolating the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid from step (a1) and A process is provided that includes.

[0033] In one embodiment (Embodiment J1), the present invention is a process for preparing the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, comprising

[0034]

Chemical formula

[0035] In one embodiment (Embodiment K1), the present invention is a process for preparing the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid,

[0036] [Chemical formula] (a1) Reacting methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with barium hydroxide [Ba(OH)2] in a solvent system comprising acetone and water to form the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, where the amount of barium hydroxide [Ba(OH)2] is about 0.5 to about 0.6 (e.g., about 0.5) molar equivalents relative to methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, (a2) Water, a water-immiscible organic solvent (e.g., toluene, TBME, or ethyl acetate), and an organic acid (e.g., acetic acid) are added to the reaction mixture from step (a1), and the resulting mixture is mixed to form (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, and (a3) Optionally, separating the organic phase from the aqueous phase of the mixture from step (a2) and adding methanol to the separated organic phase, and (a4) 2-Amino-2-(hydroxymethyl)-1,3-propanediol is added to the separated organic phase from step (a3), whereby 2-amino-2-(hydroxymethyl)-1,3-propanediol and (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid are reacted to form the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, where the reaction mixture is held at a holding temperature of from about 35 °C to about 55 °C (e.g., from about 40 °C to about 50 °C, or about 45 °C), and (a6) A seed crystalline material of the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid is added to the reaction mixture from step (a4) to form a slurry, where the slurry is held at the holding temperature for a time longer than about 1 minute, and (a7) The slurry from step (a6) is cooled to a temperature of from about 15 °C to about 20 °C (e.g., 20 °C) and the slurry is held at that temperature for a time longer than about 1 minute, and (a7) Step of isolating the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid from the slurry in step (a6) A process is provided which comprises.

[0037] In one embodiment (Embodiment L1), the present invention is a method for preparing Form 1 of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, comprising: (a) Suspending the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid in a solvent system, wherein the solvent system consists of dimethyl sulfoxide (DMSO) and water, the volume ratio of DMSO:water is from about 10:1 to about 6:1 (e.g., from about 9:1 to about 7:1, or about 8:1), and the ratio of water (volume) to the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (weight) is from about 0.8 mL / g to about 1.2 mL / g (e.g., 1.0 mL / g) at a temperature from about 20 °C to about 35 °C (e.g., from about 20 °C to about 30 °C); (b) Heating the suspension in step (a) to a temperature increase from about 60 °C to about 70 °C (e.g., about 65 °C) to form a solution, and then mixing the solution for a time longer than about 1 minute during the temperature increase; (c) Slowly add water to the solution from step (b) while maintaining the reaction mixture as a solution, where the amount of water added is approximately the same as the water used in step (a), and then hold the resulting solution at an elevated temperature for a time longer than about 1 minute, (d) Seed the solution with the crystalline form 1 material of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid while maintaining the temperature at an elevated level, where the amount of seed crystal crystallinity is at least about 0.5 wt% (e.g., about 0.5 wt% or about 1.0 wt%) of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid used in step (a), and then hold the resulting mixture at an elevated temperature for a time longer than about 1 minute, (e) Optionally, slowly cool the temperature of the mixture from step (d) to about 30 °C and then hold the mixture at that temperature for a time longer than about 1 minute, (f) Optionally, slowly heat the mixture from step (e) from about 40 °C to about 50 °C and then hold the mixture at that temperature for a time longer than about 1 minute, (g) Slowly cool the temperature of the mixture from step (d) or from step (f) (if steps (e) and (f) are carried out) to about 15 °C and then hold the mixture at that temperature for a time longer than about 1 minute, (h) Isolate the solid from the resulting mixture from step (g) to obtain the form I of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid A method is provided that includes the above steps.

[0038] In one embodiment (Embodiment M1), the present invention is a method for preparing Form 1 of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, comprising: (a) suspending the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid in a solvent system, wherein the solvent system consists of tetrahydrofuran (THF) and water, the volume ratio of THF:water is from about 1:1 to about 4:1 (e.g., from about 2.5:1 to about 3.5:1, from about 2.8:1 to about 3.2:1, from about 2.9:1 to about 3.1:1, or about 3.0:1), and the ratio of water (volume) to the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (weight) is from about 1.1 mL / g to about 3.8 mL / g (e.g., from about 1.3 mL / g to about 1.5 mL / g, or about 1.39 mL / g) at a temperature from about 20 °C to about 35 °C (e.g., from about 20 °C to about 30 °C, about 22 °C); (b) heating the suspension in step (a) to a high temperature from about 49 °C to about 59 °C (e.g., about 55 °C) to form a solution, cooling the temperature to a holding temperature from about 47 °C to about 51 °C (e.g., about 49 °C), during which the mixture remains a solution, and optionally mixing the solution at the holding temperature for a time longer than about 1 minute; (c) To the solution from step (b), crystalline form 1 material of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid is seeded while maintaining the temperature at the holding temperature, where the amount of the seed crystal crystallinity is about 0.5% by weight or more (e.g., about 0.5% by weight or about 1.0% by weight) of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid used in step (a), and then the resulting mixture is held at the holding temperature for a time longer than about 1 minute, (d) Slowly cooling the temperature of the mixture from step (c) to an intermediate temperature of about 35 °C, and then holding the mixture at the intermediate temperature for a time longer than about 1 minute, (e) A water-miscible organic solvent (e.g., acetonitrile, isopropanol, or acetone) is slowly added to the mixture from step (d) while maintaining the temperature of the mixture at the intermediate temperature, and then the mixture is held at the intermediate temperature for a time longer than about 1 minute, (f) Slowly cooling the temperature of the mixture from step (e) to a low temperature of about 10 °C, and then holding the mixture at the low temperature for a time longer than about 1 minute, (g) Taking a sample of the mixture (slurry) and determining the particle size of the solid in the mixture for a time longer than about 1 minute, (h) Performing high-shear wet milling until the D90 of the particle size of the solid in the mixture is less than about 150 μM, (i) Isolating the solid from the resulting mixture from step (f) to obtain Form I of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid A method is provided that includes the above steps.

Brief Description of the Drawings

[0039]

Figure 1

Figure 2

Figure 3

Figure 4

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Figure 6

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Figure 8

Figure 9

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Figure 14

DETAILED DESCRIPTION OF THE INVENTION

[0040] The present invention can be more easily understood by referring to the following detailed description of exemplary embodiments of the present invention and the examples included therein.

[0041] It should be understood that the present invention is not limited to specific preparation methods that can naturally vary. It should also be understood that the terms used in this specification are for the sole purpose of describing specific embodiments and are not intended to be limiting.

[0042] Any solid form of the present invention can be substantially pure. As used herein, the term "substantially pure" with respect to a particular solid form (e.g., a crystalline form) means that the particular solid form (e.g., a crystalline form) contains less than 15 wt%, less than 10 wt%, less than 5 wt%, less than 3 wt%, or less than 1 wt% of any other physical form of the tris salt of C111.

[0043] The term "substantially the same", when used to describe an X-ray powder diffraction pattern, is intended to include patterns in which the peaks (with respect to 2θ) are within the deviations specified herein.

[0044] The term "substantially the same", when used to describe an ssNMR spectrum, is intended to include ssNMR spectra in which the peaks (with respect to chemical shift) are within the deviations specified herein.

[0045] The term "substantially the same", when used to describe an FT-Raman spectrum, is intended to include an FT-Raman spectrum in which the peaks (with respect to wavenumber) are within the deviations specified herein.

[0046] The term "about" generally means within 10%, preferably within 5%, more preferably within 1% of a given value or range. Alternatively, the term "about" means within the acceptable standard error as would be considered by one of ordinary skill in the art.

[0047] The term "tris" means 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine, tromethamine, or 2-amino-2-(hydroxymethyl)propane-1,3-diol, also known as THAM.

[0048] The tris salt of C111 means a salt of C111 prepared using 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine and C111. Tris is related to the carboxylic acid moiety of C111. Unless otherwise stated, when referring to the tris salt of C111, the counterion and C111 are in a stoichiometric ratio of about 1:1 (i.e., from 0.9:1.0 to 1.0:0.9, for example, from 0.95:1.00 to 1.00:0.95). Another chemical name representing the tris salt of C111 is 1,3-dihydroxy-2-(hydroxymethyl)propan-2-aminium 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate, which can also be represented, for example, by one of the following structures.

[0049]

Chemical Structure

[0050] One of ordinary skill in the art will readily understand that multiple nomenclatures may be used to name the same compound (including the same salt).

[0051] Any example or embodiment of the solid form of the present invention can be grouped, individually or together in any combination with any number and any embodiment described herein, and can be claimed.

[0052] Room temperature (RT) or ambient temperature: 15 to 25 °C.

[0053] Dimethyl sulfoxide: DMSO.

[0054] 1 The 1H nuclear magnetic resonance (NMR) spectra were consistent with the proposed structures in all cases. Characteristic chemical shifts (δ) are reported in parts per million relative to the residual proton signal in the deuterated solvent (CHCl3 at 7.27 ppm, CD2HOD at 3.31 ppm, MeCN at 1.94 ppm, DMSO at 2.50 ppm), and the conventional abbreviations for the names of the major peaks are used: for example, s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad. The symbol ^ indicates that the peak was partially obscured by the water peak, 1 The 1H NMR peak area indicates that it was estimated. The symbol ^^ indicates that the peak was partially obscured by the solvent peak, 1 The 1H NMR peak area indicates that it was estimated.

[0055] The compounds and intermediates described hereinafter were named using the naming rules provided by ACD / ChemSketch 2012, ChemDraw, File Version C10H41, Build 69045 (Advanced Chemistry Development, Inc., Toronto, Ontario, Canada). The naming rules provided by ACD / ChemSketch 2012 are well-known to those skilled in the art, and the naming rules provided by ACD / ChemSketch 2012 are generally considered to be in compliance with the IUPAC (International Union of Pure and Applied Chemistry) recommendations and CAS index rules for the nomenclature of organic compounds. You will notice that the chemical name may have only round brackets or may have both round brackets and square brackets. The stereochemical descriptors may be placed in different locations within the name itself depending on the naming rules. Those skilled in the art will recognize these variations in format and understand that they provide the same chemical structure.

[0056] Pharmaceutically acceptable salts include acid addition and base salts.

[0057] Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, 1,5-naphthalenedisulfonic acid and xinafoate.

[0058] Suitable base salts are formed from bases that form non-toxic salts. Examples include aluminum salts, arginine salts, benzathine salts, calcium salts, choline salts, diethylamine salts, bis(2-hydroxyethyl)amine (diolamine) salts, glycine salts, lysine salts, magnesium salts, meglumine salts, 2-aminoethanol (olamine) salts, potassium salts, sodium salts, 2-amino-2-(hydroxymethyl)propane-1,3-diol (tris or tromethamine) salts, and zinc salts.

[0059] Hemisalts of acids and bases, such as hemisulfates and hemicalcium salts, can also be formed. For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use, by Stahl and Wermuth (Wiley-VCH, 2002).

[0060] Pharmaceutically acceptable salts can be prepared in three ways: (i) by reacting the compound with the desired acid or base, (ii) by removing an acid- or base-labile protecting group from a suitable precursor of the compound, or by ring-opening a suitable cyclic precursor, such as a lactone or lactam, using the desired acid or base, or (iii) by converting one salt of the compound to another by reaction with a suitable acid or base or by utilizing a suitable ion-exchange column by one or more of the above methods.

[0061] All three reactions are typically carried out in solution. The resulting salt may be condensed and collected by filtration or recovered by evaporation of the solvent. The degree of ionization in the resulting salt can vary from almost completely ionized to almost non-ionized.

[0062] Compounds and pharmaceutically acceptable salts can exist in unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex comprising the compound or a salt thereof and one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term "hydrate" is used when the solvent is water. Forms 1 and 2 described herein are believed to be unsolvated (and thus anhydrous).

[0063] The currently approved classification system for organic hydrates defines isolated-site, channel, or metal-ion coordinated hydrates; see Polymorphism in Pharmaceutical Solids, by K.R. Morris (edited by H.G. Brittain, Marcel Dekker, 1995). Isolated-site hydrates are those in which water molecules are isolated from direct contact with each other by intervening organic molecules. In channel hydrates, the water molecules are in lattice channels where they are adjacent to other water molecules. In metal-ion coordinated hydrates, the water molecules are bound to metal ions.

[0064] When the solvent or water is tightly bound, the complex can have a well-defined stoichiometry independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water / solvent content can depend on humidity and drying conditions. In such cases, non-stoichiometry is the norm.

[0065] Multicomponent complexes (other than salts and solvates) in which the drug and at least one other component are present in stoichiometric or non-stoichiometric amounts are also included within the scope of the present invention. This type of complex includes clathrates (drug-host inclusion complexes) and co-crystals. The latter are typically defined as crystalline complexes of neutral molecular components held together via non-covalent interactions, but can also be complexes of neutral molecules and salts. Co-crystals may be prepared by melt crystallization, recrystallization from a solvent, or by physically grinding the components together; see Chem Commun, 17, 1889-1896, by O. Almarsson and M. J. Zaworotko (2004). For a general review of multicomponent complexes, see J Pharm Sci, 64(8), 1269-1288, by Haleblian (August 1975).

[0066] The compounds of the present invention can exist in a series of solid states ranging from completely amorphous to completely crystalline. The term "amorphous" refers to a state in which the material lacks long-range order at the molecular level and can exhibit the physical properties of a solid or a liquid depending on temperature. Typically, such materials do not give a distinct X-ray diffraction pattern and are described more formally as liquids while exhibiting the properties of a solid. Upon heating, a change in state, typically a secondary ("glass transition") characterized change from solid to liquid-like properties, appears. The term "crystalline" refers to a solid phase in which the material has a regular ordered internal structure at the molecular level and gives a distinct X-ray diffraction pattern with defined peaks. Such materials will also exhibit the properties of a liquid when heated sufficiently, but the change from solid to liquid is characterized by a phase change, typically a primary ("melting point").

[0067] When subjected to suitable conditions, the compound may exist in an intermediate state (mesophase or liquid crystal). The intermediate state is an intermediate between the true crystalline state and the true liquid state (either molten or in solution). The liquid crystallinity resulting from a change in temperature is described as "thermotropic", and that resulting from the addition of a second component such as water or another solvent is described as "lyotropic". Compounds having the potential to form lyotropic mesophases are described as "amphiphilic" and consist of molecules having ionic (-COO - Na + ,-COO - K + or -SO3 - Na + etc.) or non-ionic (-N - N + (CH3)3 etc.) polar head groups. For further information, see Crystals and the Polarizing Microscope, by N.H. Hartshorne and A. Stuart, 4th edition (Edward Arnold, 1970).

[0068] Some compounds may exhibit polymorphism and / or one or more types of isomerism (e.g., optical, geometric or tautomeric isomerism). The solid forms of the present invention (e.g., crystalline and / or amorphous forms) may be isotopically labeled. Such variations are implicit in C111 or its salts as defined with reference to their structural characteristics and are therefore within the scope of the present invention.

[0069] Compounds containing one or more asymmetric carbon atoms can exist as two or more stereoisomers. When the compound contains an alkenyl or alkenylene group, geometric cis / trans (or Z / E) isomers are possible. Tautomeric isomerism ("tautomerism") can occur when the structural isomers are interconvertible via a low energy barrier. This can take the form of proton tautomerism, for example, in compounds containing an imino, keto, or oxime group in an aromatic moiety, or so-called valence tautomerism. It follows that a single compound can exhibit multiple types of isomerism.

[0070] Certain pharmaceutically acceptable salts of C111 may also contain counterions that are optically active (e.g., d-lactic acid or l-lysine) or racemic (e.g., dl-tartaric acid or dl-arginine).

[0071] Cis / trans isomers can be separated by conventional techniques well known to those skilled in the art, such as chromatography and fractional crystallization.

[0072] Conventional techniques for the preparation / isolation of individual enantiomers involve chiral synthesis from suitable optically pure precursors or the resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC). Alternatively, a racemic precursor containing a chiral ester can be separated by enzymatic resolution (see, for example, Int J Mol Sci 29682 - 29716, by A.C.L.M. Carvaho et al. (2015)). When the compound contains an acidic or basic moiety, salts can be formed with an optically pure base or acid such as 1-phenylethylamine or tartaric acid. The resulting mixture of diastereomers can be separated by fractional crystallization, and one or both of the diastereomeric salts can be converted to the corresponding pure enantiomers by means well known to those skilled in the art. Alternatively, the racemate (or racemic precursor) can be covalently reacted with a suitable optically active compound, such as an alcohol, an amine, or a benzyl-type chloride. The resulting mixture of diastereomers can be separated by chromatography and / or fractional crystallization by means well known to those skilled in the art to obtain the separated diastereomers as a single enantiomer having two or more chiral centers. Chiral compounds (and their chiral precursors) can be obtained in an enantiomerically enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing from 0 to 50% by volume, typically from 2 to 20% by volume, of isopropanol, and from 0 to 5% by volume, typically 0.1% diethylamine, of an alkylamine. Concentration of the eluate yields an enriched mixture. Chiral chromatography using subcritical or supercritical fluids can be used. Methods for chiral chromatography useful in some embodiments of the present invention are known in the art (see, for example, Smith, Roger M., Loughborough University, Loughborough, UK; Chromatographic Science Series (1998), 75 (SFC with Packed Columns), pp. 223 - 249 and references cited therein).In some related examples herein, the columns were obtained from Chiral Technologies, Inc, West Chester, Pennsylvania, USA, a subsidiary of Daicel (R) Chemical Industries, Ltd., Tokyo, Japan.

[0073] When any racemate crystallizes, two different types of crystals are possible. The first type is the above-mentioned racemic compound (true racemate) in which one homogeneous form of crystals containing both enantiomers in equimolar amounts is produced. The second type is a racemic mixture or conglomerate in which two forms of crystals each containing a single enantiomer are produced in equimolar amounts. All the crystal forms present in the racemic mixture have the same physical properties, but they may have different physical properties compared to the true racemate. The racemic mixture may be separated by conventional techniques known to those skilled in the art, see, for example, Stereochemistry of Organic Compounds, by E.L. Eliel and S.H. Wilen (Wiley, 1994).

[0074] It should be emphasized that C111 and its salts are depicted herein in a single tautomeric form, and all possible tautomeric forms are included within the scope of the present invention.

[0075] The present invention includes all pharmaceutically acceptable isotope-labeled C111 or salts thereof in which one or more atoms have been replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number that is predominant in nature but having the same number of atoms.

[0076] Examples of isotopes suitable for inclusion in the compounds of the present invention are 2 H and 3 H, etc. of hydrogen, 11 C, 13 C and 14 C, etc. of carbon, 36 Cl, etc. of chlorine, 13 N and 15 N, etc. of nitrogen, and 15O, 17 O and 18 contains oxygen isotopes such as O.

[0077] Certain specific isotope-labeled C111 or its salts, such as those incorporating radioisotopes, are useful in drug and / or substrate tissue distribution studies. Radioisotope tritium, i.e., 3 H, and carbon-14, i.e., 14 C are particularly useful for this purpose considering their ease of incorporation and immediate detection means.

[0078] Substitution with heavier isotopes such as deuterium, i.e., 2 H, can provide certain therapeutic advantages resulting from better metabolic stability, such as an increase in in vivo half-life or a reduction in the required dosage.

[0079] 11 C, 18 F, 15 O and 13 Substitution with positron-emitting isotopes such as N can be useful in positron emission tomography (PET) studies for examining substrate receptor occupancy.

[0080] Isotope-labeled compounds can generally be prepared by using appropriate isotope-labeled reagents in place of the previously used unlabeled reagents by conventional techniques known to those skilled in the art or by processes similar to those described in the accompanying examples and preparations.

[0081] The pharmaceutically acceptable solvates according to the present invention include those in which the crystallization solvent may be isotope-substituted, such as D2O, d6-acetone, d6-DMSO.

[0082] In one embodiment (Embodiment A1), the present invention is a process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile,

[0083]

Chem.

[0084] The progress of the reactions in each of steps (a1) and (b1) can be monitored by various techniques, for example, by chromatographic techniques (e.g., LC, TLC, or reverse phase HPLC). The reactions in each of steps (a1) and (b1) are carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0085] Embodiment A2 is a further embodiment of Embodiment A1, wherein the amount of the palladium catalyst [e.g., palladium(II) acetate or tris(dibenzylideneacetone)dipalladium] is from about 0.1 mol% to about 2.0 mol% (e.g., from about 0.15 mol% to about 0.5 mol%, or 0.25 mol%) of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1). In some further embodiments, the palladium catalyst is palladium(II) acetate. In some further embodiments, palladium(II) acetate is dissolved in anisole to form a solution, and then added to the reaction mixture in step (a1).

[0086] Embodiment A3 is a further embodiment of Embodiment A1 or A2, wherein the amount of the phosphorus ligand (e.g., JohnPhos) is about 2.0 molar equivalents relative to the palladium catalyst [e.g., palladium(II) acetate] in step (a1). In some further embodiments, the ligand is JohnPhos. In some further embodiments, JohnPhos is dissolved in anisole to form a solution, and then added to the reaction mixture in step (a1).

[0087] Embodiment A4 is a further embodiment of any one of Embodiments A1 to A3, wherein the amount of the base (for example, tripotassium phosphate) is about 1 to about 2 molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1). In some further embodiments, the base is tripotassium phosphate. Also in some further embodiments, the amount of tripotassium phosphate is about 1.5 to about 1.9 (for example, 1.7) molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1).

[0088] Embodiment A5 is a further embodiment of any one of Embodiments A1 to A4, wherein the volume amount of anisole is about 5 ml / g to about 10 ml / g (for example, about 7 ml / g to about 9 ml / g, or about 8 ml / g) based on the weight of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1).

[0089] Embodiment A6 is a further embodiment of any one of Embodiments A1 to A5, wherein the reaction mixture is stirred at about 80 to about 120 °C (for example, about 90 to about 110 °C, or about 100 °C) for a time sufficient to form tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (a1). The progress of the reaction can be monitored by various techniques, for example, by chromatographic techniques (such as LC, TLC, or reverse-phase HPLC). In some embodiments, the reaction takes about 16 to about 24 hours to complete.

[0090] Embodiment A7 is a further embodiment of Embodiment A6, wherein the reaction mixture is cooled to ambient temperature after completion of the reaction in step (a1) and before step (a2) is carried out.

[0091] Embodiment A8 is a further embodiment of any one of Embodiments A1 to A7, wherein the volume amount of ethanol in step (a2) is about 0.2 to about 0.3 (e.g., 0.25) equivalents of the volume amount of anisole in step (a1).

[0092] Embodiment A9 is a further embodiment of any one of Embodiments A1 to A8, wherein filtering in step (a3) further includes washing with ethyl acetate. In some further embodiments, the volume amount of ethyl acetate used for washing is about 0.4 to about 0.6 (e.g., about 0.5) equivalents of the volume amount of anisole in step (a1).

[0093] The filtrate from step (a3) is used directly in step (b1).

[0094] Embodiment A10 is a further embodiment of any one of Embodiments A1 to A9, wherein the amount of p-toluenesulfonic acid monohydrate is about 2.0 to about 2.5 molar equivalents relative to tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (b1).

[0095] Embodiment A11 is a further embodiment of any one of Embodiments A1 to A10, wherein the amount of p-toluenesulfonic acid monohydrate is about 2.1 to about 2.4 (e.g., 2.2) molar equivalents relative to tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (b1).

[0096] Embodiment A12 is a further embodiment of any one of Embodiments A1 to A11, in which the reaction mixture in step (b1) is stirred at about 30 °C to about 60 °C (for example, about 35 °C to about 50 °C, or about 40 °C) for a time sufficient to form the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile. The progress of the reaction can be monitored by various techniques, for example by chromatographic techniques (such as LC, TLC, or reverse-phase HPLC).

[0097] Embodiment A13 is a further embodiment of any one of Embodiments A1 to A12, in which p-toluenesulfonic acid monohydrate is added as an undiluted reagent (i.e., without pre-mixing with a solvent) to the reaction mixture in step (b1).

[0098] Embodiment A14 is a further embodiment of any one of Embodiments A1 to A13, in which isolating the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in step (b2) involves cooling the reaction mixture and filtering the mixture after the reaction from step (b1) is complete. In some further embodiments, cooling the reaction mixture from step (b1) involves cooling the reaction mixture to about 0 °C. Also, in some further embodiments, cooling the reaction mixture involves cooling the reaction mixture to about 0 °C with stirring over at least 1 hour.

[0099] Embodiment A15 is a further embodiment of Embodiment A14, in which filtering the mixture further involves washing the solid obtained by filtration with anisole. In some further embodiments, filtering the mixture further involves washing the solid obtained by filtration with anisole and ethyl acetate.

[0100] Embodiment A16 is a further embodiment of any one of Embodiments A1 to A15, wherein the isolated bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in step (b2) is further dried, optionally under vacuum. In some further embodiments, the vacuum drying is carried out at a temperature of about 30 °C or lower, about 35 °C or lower, about 40 °C or lower, about 45 °C or lower, about 50 °C or lower, or about 60 °C or lower. In some further embodiments, the vacuum drying is carried out at a temperature of about 40 °C or lower.

[0101] Embodiment A17 is a further embodiment of any one of Embodiments A1 to A16, wherein the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile isolated in step (b2) is in the anhydrous form.

[0102] In one embodiment (Embodiment B1), the present invention provides a process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, comprising

[0103] [Chemical formula] (a1)tert-Butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate is reacted with 3-fluoro-4-(hydroxymethyl)benzonitrile, a palladium catalyst [palladium(II) acetate, tris(dibenzylideneacetone)dipalladium, or palladium(II) chloride, etc.], a base [an inorganic base, e.g., tripotassium phosphate, cesium carbonate, potassium hydroxide, or sodium hydride, etc.], and a phosphorus ligand [a monodentate phosphorus ligand or a bidentate phosphorus ligand, e.g., 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (XPhos), (2-biphenylyl)di-tert-butylphosphine (JohnPhos), 5-(di-tert-butylphosphino)-1′,3′,5′-triphenyl-1′H-[1,4′]bipyrazole (Bippyphos), 5-[bis(1-adamantyl)phosphino]-1′,3′,5′-triphenyl-1,4′-bi-1H-pyrazole (AdBippyphos), 2-(dicyclohexylphosphino)-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl (BrettPhos), di(adamantan-1-yl)(2′,4′,6′-triisopropyl-3,6-dimethoxy-2-biphenylyl)phosphine (AdBrettPhos), 2-dicyclohexylphosphino-2′,6-bis(N,N-dimethylamino)biphenyl (CPhos), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (SPhos), 2-dicyclohexylphosphino-2′-(N,N-dimethylamino)biphenyl (DavePhos), 2-di-tert-butylphosphino-2′-(N,N-dimethylamino)biphenyl (tBuDavePhos), 2-(di-tert-butylphosphino)-2′,4′,6′-triisopropyl-3,6-dimethoxy-1,1′-biphenyl (tBuBrettPhos), di-tert-butyl(2′,4′,6′-triisopropyl-3-methoxy-6-methyl-[1,1′-biphenyl]-2-yl)phosphine (RockPhos), 4,5-bis(diphenylphosphino)-9,In the presence of 9-dimethylxanthene (XantPhos), or bis[(2-diphenylphosphino)phenyl]ether (DPEPhos), etc., in a solvent system [such as those containing anisole, acetonitrile, tetrahydrofuran, dioxane, or methyl tert-butyl ether, etc.], react to form tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate, where the amount of 3-fluoro-4-(hydroxymethyl)benzonitrile is about 1.0 to about 1.1 molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate, and, (a2) Adding water, ethyl acetate, and ethanol to the reaction mixture from step (a1); (a3) Separating the organic phase from the aqueous phase in step (a2); (b1) Adding p-toluenesulfonic acid monohydrate to the separated organic phase from step (a3), thereby reacting tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate with p-toluenesulfonic acid monohydrate to form the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, where the amount of p-toluenesulfonic acid monohydrate is about 2.0 to about 3.0 molar equivalents relative to tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate; (b2) Isolating the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile from step (b1); A process is provided that includes the above steps.

[0104] The progress of the reactions in each of steps (a1) and (b1) can be monitored by various techniques, for example by chromatographic techniques (e.g., LC, TLC, or reverse phase HPLC). The reactions in each of steps (a1) and (b1) are carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0105] Embodiment B2 is a further embodiment of Embodiment B1 where the amount of palladium catalyst [e.g., palladium(II) acetate] is from about 0.1 mol% to about 0.75 mol% (e.g., 0.25 mol%) of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1). In some further embodiments, the palladium catalyst is palladium(II) acetate. Also in some further embodiments, palladium(II) acetate is dissolved in anisole to form a solution and then added to the reaction mixture in step (a1).

[0106] Embodiment B3 is a further embodiment of Embodiment B1 or B2 where the amount of ligand (e.g., X-Phos) is about 1.0 molar equivalent relative to the palladium catalyst [e.g., palladium(II) acetate] in step (a1). In some further embodiments, the ligand is X-Phos. Also in some further embodiments, X-Phos is dissolved in anisole to form a solution and then added to the reaction mixture in step (a1).

[0107] Embodiment B4 is a further embodiment of any one of Embodiments B1 to B3, wherein the amount of the base (for example, tribasic potassium phosphate) is about 1 to about 2 molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1). In some further embodiments, the base is tribasic potassium phosphate. Also in some further embodiments, the amount of tribasic potassium phosphate is about 1.5 to about 1.9 (for example, 1.7) molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1).

[0108] Embodiment B5 is a further embodiment of any one of Embodiments B1 to B4, wherein the volume amount of anisole is about 5 ml / g to about 10 ml / g (for example, about 8 ml / g) based on the weight of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1).

[0109] Embodiment B6 is a further embodiment of any one of Embodiments B1 to B5, wherein the reaction mixture is stirred at about 100 °C for a time sufficient to form tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (a1). The progress of the reaction can be monitored by various techniques, for example, by chromatographic techniques (for example, LC, TLC, or reverse-phase HPLC). In some embodiments, the reaction takes about 16 to 24 hours to complete.

[0110] Embodiment B7 is a further embodiment of Embodiment B6, wherein the reaction mixture is cooled to ambient temperature after completion of the reaction in step (a1) and before step (a2) is carried out.

[0111] Embodiment B8 is a further embodiment of any one of Embodiments B1 to B7, wherein the volume amount of water in step (a2) is about 0.3 to about 0.45 (e.g., 0.375) equivalents of the volume amount of anisole in step (a1).

[0112] Embodiment B9 is a further embodiment of any one of Embodiments B1 to B8, wherein the volume amount of ethanol in step (a2) is about 0.2 to about 0.3 (e.g., 0.25) equivalents of the volume amount of anisole in step (a1).

[0113] Embodiment B10 is a further embodiment of any one of Embodiments B1 to B8, wherein the volume amount of ethyl acetate in step (a2) is about 0.40 to about 0.60 (e.g., 0.50) equivalents of the volume amount of anisole in step (a1).

[0114] After adding water, ethyl acetate, and ethanol in step (a2), the resulting mixture is thoroughly mixed, then the two layers are allowed to settle, and then in step (a3), the organic layer (or organic phase) is separated from the aqueous phase. The organic layer (or organic phase) separated from step (a3) is used directly in step (b1).

[0115] Embodiment B11 is a further embodiment of any one of Embodiments B1 to B10, wherein the amount of p-toluenesulfonic acid monohydrate is about 2.1 to about 2.5 molar equivalents relative to tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (b1).

[0116] Embodiment B12 is a further embodiment of any one of Embodiments B1 to B11, wherein the amount of p-toluenesulfonic acid monohydrate is about 2.1 to about 2.4 (e.g., 2.2) molar equivalents relative to tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (b1).

[0117] Embodiment B13 is a further embodiment of any one of Embodiments B1 to B12, in which the reaction mixture in step (b1) is stirred at about 40° C. for a time sufficient to form the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile. The progress of the reaction can be monitored by various techniques, for example by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC).

[0118] Embodiment B14 is a further embodiment of any one of Embodiments B1 to B13, in which p-toluenesulfonic acid monohydrate is added as an undiluted reagent (i.e., without pre-mixing with a solvent) to the reaction mixture in step (b1).

[0119] Embodiment B15 is a further embodiment of any one of Embodiments B1 to B14, in which isolating the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in step (b2) includes cooling the reaction mixture and filtering the mixture after the reaction from step (b1) is complete. In some further embodiments, cooling the reaction mixture from step (b1) includes cooling the reaction mixture to about 5° C. In some further embodiments, cooling the reaction mixture includes cooling the reaction mixture to about 5° C. over at least 1 hour while stirring the reaction mixture.

[0120] Embodiment B16 is a further embodiment of Embodiment B15, in which filtering the mixture further includes washing the solid obtained by filtration with ethyl acetate.

[0121] Embodiment B17 is a further embodiment of any one of Embodiments B1 to B16, wherein the isolated bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in step (b2) is further dried, optionally under vacuum. In some further embodiments, the vacuum drying is carried out at a temperature of about 30 °C or lower, about 35 °C or lower, about 40 °C or lower, about 45 °C or lower, about 50 °C or lower, or about 60 °C or lower. In some further embodiments, the vacuum drying is carried out at a temperature of about 40 °C or lower.

[0122] Embodiment B18 is a further embodiment of any one of Embodiments B1 to B17, wherein the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile isolated in step (b2) is the monohydrate of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0123] In one embodiment (Embodiment C1), the present invention is a process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, comprising

[0124]

Chemical formula

[0125] The progress of the reactions in each of steps (a1) and (b1) can be monitored by various techniques, such as chromatographic techniques (e.g., 1H NMR, TLC, or reverse-phase HPLC). The reactions in each of steps (a1) and (b1) are carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0126] Embodiment C2 is a further embodiment of Embodiment C1, where the amount of the copper catalyst [tetrakis(acetonitrile)copper(I) triflate, copper(I) iodide, copper(I) acetate, tetrakis(acetonitrile)copper(I) hexafluorophosphate, or copper trifluoromethanesulfonate, etc.] is from about 4.0 mol% to about 20 mol% (e.g., from about 4.0 mol% to about 6.0 mol%, or about 5.0 mol%) of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1). In some further embodiments, the copper catalyst is tetrakis(acetonitrile)copper(I) triflate.

[0127] Embodiment C3 is a further embodiment of Embodiment C1 or C2, where the amount of the ligand (e.g., N,N'-bis(1-naphthylmethyl)oxamide) is about 1.0 molar equivalent relative to the copper catalyst [tetrakis(acetonitrile)copper(I) triflate, etc.] in step (a1). In some further embodiments, the ligand is N,N'-bis(1-naphthylmethyl)oxamide.

[0128] Embodiment C4 is a further embodiment of any one of Embodiments C1 to C3, where the amount of the base (e.g., sodium tert-pentoxide) is from about 1 to about 2 molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1). In some further embodiments, the base is sodium tert-pentoxide. Also in some further embodiments, the amount of sodium tert-pentoxide is from about 1.3 to about 1.7 (e.g., 1.5) molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1). In some further embodiments, after dissolving sodium tert-pentoxide in toluene to form a solution, it is added to the reaction mixture in step (a1).

[0129] Embodiment C5 is a further embodiment of any one of Embodiments C1 to C4, wherein the solvent system in step (a1) is 1,4-dioxane, and the volume amount of 1,4-dioxane is about 7 ml / g to about 9 ml / g (for example, about 8 ml / g) based on the weight of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate in step (a1).

[0130] Embodiment C6 is a further embodiment of any one of Embodiments C1 to C5, wherein the reaction mixture is stirred at about 80 °C for a time sufficient to form tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (a1). The progress of the reaction can be monitored by various techniques, for example, by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). In some embodiments, the reaction takes about 16 to 24 hours to complete.

[0131] Embodiment C7 is a further embodiment of Embodiment C6, wherein after completion of the reaction in step (a1) and before step (a2) is carried out, the reaction mixture is cooled to ambient temperature.

[0132] Embodiment C8 is a further embodiment of any one of Embodiments C1 to C7, wherein the volume amount of MTBE in step (a2) is about 0.2 to about 0.3 (for example, 0.25) equivalents of the volume amount of the solvent 1,4-dioxane in step (a1). In some further embodiments, MTBE is divided into two portions for washing. Also, in some further embodiments, the reaction vessel is further rinsed with 1,4-dioxane [for example, about 0.2 volume equivalents of the volume amount of the solvent 1,4-dioxane in step (a1)], and filtration is also carried out before the MTBE wash (one or more times).

[0133] In step (a3), the solvent is removed, and the obtained tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate is carried over to step (b1).

[0134] Embodiment C9 is a further embodiment of any one of Embodiments C1 to C8, wherein the volume amount of 1,4-dioxane used to dissolve tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (b1) is about 1.5 to about 1.9 (e.g., about 1.67) ml / g based on the weight of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate used in step (a1).

[0135] Embodiment C10 is a further embodiment of any one of Embodiments C1 to C9, wherein the volume amount of MTBE used to dissolve tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (b1) is about 0.75 to about 0.95 (e.g., about 0.83) ml / g based on the weight of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate used in step (a1).

[0136] Embodiment C11 is a further embodiment of any one of Embodiments C1 to C10, wherein the amount of p-toluenesulfonic acid monohydrate is about 2.0 to about 2.2 (e.g., about 2.1) molar equivalents relative to tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate in step (b1), assuming a 100% reaction yield from step (a1).

[0137] Embodiment C12 is a further embodiment of any one of Embodiments C1 to C11, wherein p-toluenesulfonic acid monohydrate is added to the reaction mixture in step (b1) as an undiluted reagent (i.e., without pre-mixing with a solvent).

[0138] Embodiment C13 is a further embodiment of any one of Embodiments C1 to C12, wherein the reaction mixture in step (b1) is stirred at about 40 °C for a time sufficient to form the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile. The progress of the reaction can be monitored by various techniques, for example by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). In some further embodiments, additional 1,4-dioxane is added to collect the reaction slurry. The volume amount of the additional 1,4-dioxane is about 3.0 to about 3.8 (e.g., about 3.3) ml / g based on the weight of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate used in step (a1).

[0139] Embodiment C14 is a further embodiment of any one of Embodiments C1 to C13, wherein isolating the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in step (b2) includes cooling the reaction mixture and filtering the mixture after the reaction from step (b1) is complete. In some further embodiments, cooling the reaction mixture from step (b1) includes cooling the reaction mixture to about 0 - 5 °C. Also in some further embodiments, cooling the reaction mixture includes cooling the reaction mixture to about 0 °C with stirring over at least about 1 hour.

[0140] Embodiment C15 is a further embodiment of Embodiment C14, wherein filtering the mixture further comprises washing the solid obtained by filtration with a mixture of 1,4-dioxane:MTBE (1:1 by volume). The amount of the 1,4-dioxane:MTBE washing solution is about 3.0 to about 3.8 (e.g., about 3.3) ml / g based on the weight of tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate used in step (a1).

[0141] Embodiment C16 is a further embodiment of any one of Embodiments C1 to C15, wherein the isolated bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in step (b2) is further dried, optionally under vacuum. In some further embodiments, the vacuum drying is carried out at a temperature of about 30 °C or lower, about 35 °C or lower, about 40 °C or lower, about 45 °C or lower, about 50 °C or lower, or about 60 °C or lower. In some further embodiments, the vacuum drying is carried out at a temperature of about 40 °C or lower.

[0142] Embodiment C17 is a further embodiment of any one of Embodiments C1 to C16, wherein the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile isolated in step (b2) is in the anhydrous form of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0143] In one embodiment (Embodiment D1), the present invention provides anhydrous 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt, The monohydrate of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The monotosylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The bismesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The monomesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The monosulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, The hemisulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, Crystalline methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, or (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid hemibarium salt which provides an intermediate useful for preparing C111 or the tris salt of C111.

[0144] Embodiment D2 is a further embodiment of Embodiment D1, wherein the present invention provides an anhydrous crystalline form of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0145] Embodiment D2A is a further embodiment of Embodiment D2, wherein the anhydrous crystalline form is Form 1 of anhydrous 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt, and Form 1 has a powder X-ray diffraction pattern (PXRD) including at least two peaks selected from those at 13.3 ± 0.2°, 15.5 ± 0.2°, and 17.2 ± 0.2° with respect to 2θ.

[0146] Embodiment D2A1 is a further embodiment of Embodiment D2A, wherein Form 1 has a PXRD including at least three peaks at 13.3 ± 0.2°, 15.5 ± 0.2°, and 17.2 ± 0.2° with respect to 2θ.

[0147] Embodiment D2A2 is a further embodiment of Embodiment D2A, wherein Form 1 has a PXRD including at least two peaks selected from those at 12.8 ± 0.2°, 13.3 ± 0.2°, 14.7 ± 0.2°, 15.5 ± 0.2°, and 17.2 ± 0.2° with respect to 2θ.

[0148] Embodiment D2A3 is a further embodiment of Embodiment D2A, wherein Form 1 has a PXRD including at least three peaks selected from those at 12.8 ± 0.2°, 13.3 ± 0.2°, 14.7 ± 0.2°, 15.5 ± 0.2°, and 17.2 ± 0.2° with respect to 2θ.

[0149] Embodiment D2A4 is a further embodiment of Embodiment D2A, wherein Form 1 has a PXRD including peaks at 12.8 ± 0.2°, 13.3 ± 0.2°, 14.7 ± 0.2°, 15.5 ± 0.2°, and 17.2 ± 0.2° with respect to 2θ.

[0150] Embodiment D2A5 is a further embodiment of Embodiment D2A, wherein Form 1 has a PXRD substantially as shown in Figure 1.

[0151] Embodiment D3 is a further embodiment of Embodiment D1, wherein the present invention provides a monohydrate crystalline form of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt.

[0152] Embodiment D3A is a further embodiment of Embodiment D3, wherein the monohydrate crystalline form is Form 2 of the monohydrate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt, and Form 2 has a powder X-ray diffraction pattern (PXRD) comprising at least two peaks selected from those at 2θ of 13.0±0.2°, 13.7±0.2°, and 22.7±0.2°.

[0153] Embodiment D3A1 is a further embodiment of Embodiment D3A, wherein Form 2 has a PXRD comprising at least three peaks at 2θ of 13.0±0.2°, 13.7±0.2°, and 22.7±0.2°.

[0154] Embodiment D3A2 is a further embodiment of Embodiment D3A, wherein Form 2 has a PXRD comprising at least two peaks selected from those at 2θ of 13.0±0.2°, 13.7±0.2°, 17.0±0.2°, 22.7±0.2°, and 27.9±0.2°.

[0155] Embodiment D3A3 is a further embodiment of Embodiment D3A, wherein Form 2 has a PXRD comprising at least three peaks selected from those at 2θ of 13.0±0.2°, 13.7±0.2°, 17.0±0.2°, 22.7±0.2°, and 27.9±0.2°.

[0156] Embodiment D3A4 is a further embodiment of Embodiment D3A, wherein Form 2 has a PXRD that includes peaks at 13.0 ± 0.2°, 13.7 ± 0.2°, 17.0 ± 0.2°, 22.7 ± 0.2°, and 27.9 ± 0.2° with respect to 2θ.

[0157] Embodiment D3A5 is a further embodiment of Embodiment D3A, wherein Form 2 has a PXRD substantially as shown in Figure 2.

[0158] Embodiment D4 is a further embodiment of Embodiment D1, wherein the present invention provides the monotosylate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0159] Embodiment D4A is a further embodiment of Embodiment D4, wherein the monotosylate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is crystalline.

[0160] Embodiment D4A1 is a further embodiment of Embodiment D4A, wherein the crystalline monotosylate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile has a PXRD substantially as shown in Figure 3.

[0161] Embodiment D5 is a further embodiment of Embodiment D1, wherein the present invention provides the bismesylate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0162] Embodiment D5A is a further embodiment of Embodiment D5, wherein the bismesylate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is crystalline.

[0163] Embodiment D5A1 is a further embodiment of Embodiment D5A, wherein the crystalline bis(mesylate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile has a PXRD substantially as shown in Figure 4.

[0164] Embodiment D6 is a further embodiment of Embodiment D1, wherein the present invention provides a monomesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0165] Embodiment D6A is a further embodiment of Embodiment D6, wherein the monomesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is crystalline.

[0166] Embodiment D6A1 is a further embodiment of Embodiment D6A, wherein the crystalline monomesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile has a PXRD substantially as shown in Figure 5.

[0167] Embodiment D7 is a further embodiment of Embodiment D1, wherein the present invention provides a monosulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0168] Embodiment D7A is a further embodiment of Embodiment D7, wherein the monosulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is crystalline.

[0169] Embodiment D7A1 is a further embodiment of Embodiment D7A, wherein the crystalline monosulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile has a PXRD substantially as shown in Figure 6.

[0170] Embodiment D8 is a further embodiment of Embodiment D1, in which the present invention provides the hemisulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile.

[0171] Embodiment D8A is a further embodiment of Embodiment D8, in which the hemisulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is crystalline.

[0172] Embodiment D8A1 is a further embodiment of Embodiment D8A, in which the crystalline hemisulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile has a PXRD substantially as shown in Figure 7.

[0173] Embodiment D9 is a further embodiment of Embodiment D1, in which the present invention provides crystalline methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0174] Embodiment D9A is a further embodiment of Embodiment D9, in which crystalline methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate is in Form X, and Form X has a powder X-ray diffraction pattern (PXRD) including at least two peaks selected from those at 2θ of 8.4.0 ± 0.2°, 13.7 ± 0.2°, and 15.0 ± 0.2°.

[0175] Embodiment D9A1 is a further embodiment of Embodiment D9A, in which Form X has a PXRD including at least three peaks at 2θ of 8.4.0 ± 0.2°, 13.7 ± 0.2°, and 15.0 ± 0.2°.

[0176] Embodiment D9A2 is a further embodiment of Embodiment D9A, wherein Form X has a PXRD comprising at least two peaks selected from those at 2θ of 8.4.0 ± 0.2°, 11.9 ± 0.2°, 13.7 ± 0.2°, and 15.0 ± 0.2°, and 19.2 ± 0.2°.

[0177] Embodiment D9A3 is a further embodiment of Embodiment D9A, wherein Form X has a PXRD comprising at least three peaks selected from those at 2θ of 4.0 ± 0.2°, 11.9 ± 0.2°, 13.7 ± 0.2°, and 15.0 ± 0.2°, and 19.2 ± 0.2°.

[0178] Embodiment D9A4 is a further embodiment of Embodiment D9A, wherein Form X has a PXRD comprising peaks at 2θ of 4.0 ± 0.2°, 11.9 ± 0.2°, 13.7 ± 0.2°, and 15.0 ± 0.2°, and 19.2 ± 0.2°.

[0179] Embodiment D9A5 is a further embodiment of Embodiment D9A, wherein Form X has a PXRD substantially as shown in Figure 14.

[0180] Embodiment D10 is a further embodiment of Embodiment D1, wherein the present invention provides the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, and its structure is shown below.

[0181] [Chemical formula]

[0182] Embodiment D10A1 is a further embodiment of Embodiment D10, wherein the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid is crystalline.

[0183] Embodiment D10A2 is a further embodiment of Embodiment D10A1, wherein the crystalline hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid has a powder X-ray diffraction pattern (PXRD) substantially as shown in Figure 8.

[0184] In one embodiment (Embodiment E1), the present invention provides a process for preparing methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, comprising:

[0185] [Chemical formula] (a1) preparing methyl (S)-4-amino-3-(oxetane-2-carboxamide)benzoate; (a2) reducing methyl (S)-4-amino-3-(oxetane-2-carboxamide)benzoate in the presence of a reducing reagent to form methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate; and (a3) reacting methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate with 2-chloro-1,1,1-trimethoxyethane in the presence of an acid (e.g., an organic acid such as citric acid or p-toluenesulfonic acid) to form methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. The process is provided.

[0186] The progress of the reactions in steps (a2) and (a3) can be monitored by various techniques, for example, by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). The reactions in steps (a2) and (a3) are carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0187] In some further embodiments of embodiment E1, methyl (S)-4-amino-3-(oxetane-2-carboxamide) benzoate can be prepared by a method similar to that described in Example 5 below. Ethyl (S)-oxetane-2-carboxylate is hydrolyzed in the presence of a base (e.g., an alkali metal hydroxide, such as an inorganic strong base like KOH or NaOH) to form a salt such as the potassium salt of (S)-oxetane-2-carboxylic acid. The potassium salt of (S)-oxetane-2-carboxylic acid can be treated with triethylamine hydrochloride to form the triethylamine salt of (S)-oxetane-2-carboxylic acid. The triethylamine salt of (S)-oxetane-2-carboxylic acid can then be reacted with methyl 3,4-diaminobenzoate in the presence of a base (e.g., diisopropylethylamine) to form methyl (S)-4-amino-3-(oxetane-2-carboxamide) benzoate.

[0188] Embodiment E2 is a further embodiment of embodiment E1, wherein the reducing reagent in step (a2) comprises a metal borohydride such as lithium borohydride or sodium borohydride. In some further embodiments, the reducing reagent comprises lithium borohydride. Also in some further embodiments, the amount of the metal borohydride is about 1.0 to about 1.5 (e.g., about 1.25) molar equivalents relative to the amount of methyl (S)-4-amino-3-(oxetane-2-carboxamide) benzoate in step (a2).

[0189] Embodiment E3 is a further embodiment of Embodiment E1 or E2, wherein the reducing reagent in step (a2) contains a metal borohydride such as lithium borohydride or sodium borohydride, and the reduction reaction is carried out in the presence of a borate ester compound (e.g., a trialkyl borate such as triethyl borate). In some embodiments, the amount of the borate ester (e.g., triethyl borate) is about 2.5 to about 3.5 (e.g., 3.0) molar equivalents relative to the amount of methyl (S)-4-amino-3-(oxetan-2-carboxamido)benzoate in step (a2).

[0190] Embodiment E4 is a further embodiment of any one of Embodiments E1 to E3, wherein the reducing reagent in step (a2) contains a metal borohydride such as lithium borohydride or sodium borohydride, and the reduction reaction is carried out in the presence of a borate ester compound (e.g., a trialkyl borate such as triethyl borate). In some embodiments, the amount of the borate ester (e.g., triethyl borate) is about 2.5 to about 3.5 (e.g., 3.0) molar equivalents relative to the amount of methyl (S)-4-amino-3-(oxetan-2-carboxamido)benzoate in step (a2).

[0191] The progress of the reaction in step (a2) can be monitored by various techniques, for example, by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). The reaction can be carried out using a suitable organic solvent such as dichloromethane. The reaction is carried out at a suitable temperature, for example, from about 10°C to about 35°C (e.g., 20°C or room temperature / ambient temperature). The work-up can be carried out by quenching the reaction mixture (after completion of the reaction) with water and separating the organic phase from the aqueous phase. The organic phase can be washed with an acidic aqueous solution [e.g., a phosphoric acid solution (0.5 M) and / or a citric acid aqueous solution (0.5 M)].

[0192] The organic phase containing methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate is concentrated by evaporating the solvent (for example, under vacuum to remove most or all of the solvent). The residue can be used in step (a3).

[0193] Embodiment E5 is a further embodiment of any one of embodiments E1 to E4, wherein the amount of 2-chloro-1,1,1-trimethoxyethane in step (a3) is about 1.00 to 1.1 (for example, 1.02 to about 1.07, such as 1.05) molar equivalents relative to the amount of methyl (S)-4-amino-3-(oxetane-2-carboxamide)benzoate in step (a2).

[0194] Embodiment E6 is a further embodiment of any one of embodiments E1 to E5, wherein the reaction in step (a3) is carried out in a solvent system containing isopropanol.

[0195] Embodiment E7 is a further embodiment of any one of embodiments E1 to E6, wherein the amount of acid in step (a3) is less than about 1.5 mol% equivalent (or less than 1.2 mol% or about 1.0 mol%) of the amount of methyl (S)-4-amino-3-(oxetane-2-carboxamide)benzoate in step (a2). In some further embodiments, the acid is citric acid.

[0196] Embodiment E8 is a further embodiment of any one of embodiments E1 to E7, wherein the reaction in step (a3) is carried out at about 40°C to about 60°C (for example, 50°C).

[0197] Embodiment E9 is a further embodiment of any one of embodiments E1 to E8, further comprising isolating methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate formed in step (a3).

[0198] The reaction in step (a3) can be carried out as follows [when the solvent contains or is isopropanol (IPA)]. After completion of the reaction, the reaction mixture is cooled to about 30 °C or lower. Then, water is added to form a suspension. The suspension is then cooled to about 5 °C, followed by filtration and washing with IPA / water (e.g., 1:9 volume ratio). The solid collected by filtration is dried, for example, under vacuum and at a temperature of about 30 °C to 50 °C (e.g., 40 °C).

[0199] In one embodiment (Embodiment F1), the present invention is a process for preparing methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, comprising

[0200] [Chemical formula] (a1) Methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate is reacted with the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in a solvent system containing acetonitrile in the presence of diisopropylethylamine to form methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, wherein the amount of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is from about 1.1 to about 1.5 molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and the amount of diisopropylethylamine is from about 4.0 to about 6.0 (e.g., 5.0) molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. A process is provided that includes the above.

[0201] The progress of the reaction in step (a1) can be monitored by various techniques, such as chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). The reaction in step (a1) is carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0202] Embodiment F2 is a further embodiment of Embodiment F1, wherein the amount of the solvent system is from about 7 ml / g to about 9 ml / g (e.g., 8 ml / g) based on the amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0203] Embodiment F3 is that the process (b1) adding the total amount of the solvent system to the reactor, where the solvent system is maintained at a temperature of about 20 °C to about 30 °C (e.g., 25 °C, designated as the "holding temperature"), and the amount of the solvent system is about 8 ml / g based on the amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate; (b2) adding a portion (e.g., 1 / 2) of the total amount of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile to the reactor while maintaining the temperature within 5 °C of the holding temperature in step (b1); (b3) adding a portion of the total amount of diisopropylethylamine to the reactor over a period of time while maintaining the temperature within 5 °C of the holding temperature in step (b1), where the portion of the total amount of diisopropylethylamine is the same as the portion of the total amount of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in step (b2); (b4) repeating steps (b2) and (b3) until the total amounts of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile and diisopropylethylamine are added to the reactor; (b5) adding the total amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate to the reactor; (b6) heating the reaction mixture to a temperature of about 45 °C to about 60 °C (e.g., 50 °C ± 5.0 °C) and maintaining the reaction mixture at that temperature for a sufficient time to form methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. (b7) Seeding the reaction mixture with seed crystal material of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, wherein the amount of the seed crystal material is about 1% by weight of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and after seeding, maintaining the reaction mixture at a temperature of about 45 °C to about 60 °C for a time longer than about 1 minute; (b8) Adding water [wherein the amount of water is about 11 ml / g to about 13 ml / g (e.g., 12 ml / g) based on the amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate] to the reaction mixture over a period of time while maintaining the temperature at about 40 °C to about 55 °C; (b9) Cooling the reaction mixture to 15 °C ± 5.0 °C over about 3 hours (at a rate of about 0.2 °C / min); (b10) Stirring the slurry resulting from step (b9) at 15 °C ± 5.0 °C for at least 5 hours (e.g., at least 8 hours); (b11) Filtering the slurry from step (b10); (b12) Washing the reaction vessel with water and acetonitrile, wherein the washing liquid is cooled to 15 °C ± 5.0 °C; (b13) Transferring the washing liquid from step (b12) to a filter; (b14) Collecting the solid from the filtrate [step (b13)]; (b15) Optionally drying the collected solid under vacuum at about 45 °C to about 55 °C (e.g., 50 °C) which is a further embodiment of Embodiment F1 or F2.

[0204] In one embodiment (Embodiment G1), the present invention is a process for preparing methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate,

[0205]

Chemical formula

[0206] The progress of the reaction in step (a1) can be monitored by various techniques, for example, by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). The reaction in step (a1) is carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0207] Embodiment G2 is a further embodiment of Embodiment G1, wherein the amount of the solvent system is about 11 ml / g to about 13 ml / g (e.g., 12 ml / g) based on the amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0208] Embodiment G3 is such that the process (b1) adding the total amount of the solvent system to the reactor, where the solvent system is maintained at a temperature of about 20 °C to about 30 °C (e.g., 25 °C, designated as the "holding temperature"), and the amount of the solvent system is about 11 ml / g to about 13 ml / g (e.g., 12 ml / g) based on the amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate; and (b2) adding the total amount of bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile to the reactor while maintaining the temperature within 5 °C of the holding temperature in step (b1); and (b3) adding the total amount of diisopropylethylamine to the reactor over a period of time while maintaining the temperature within 5 °C of the holding temperature in step (b1); and (b4) adding the total amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate to the reactor. (b5) Heating the reaction mixture to a temperature of about 45 °C to about 60 °C (e.g., 50 °C ± 5.0 °C) and maintaining the reaction mixture at that temperature for a sufficient time to form methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate; (b6) Seeding the reaction mixture with seed crystal material of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, wherein the amount of the seed crystal material is about 1% by weight of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and after seeding, maintaining the reaction mixture at a temperature of about 45 °C to about 60 °C for a time longer than about 1 minute; (b7) Adding water [wherein the amount of water is about 3 ml / g to about 5 ml / g (e.g., 4 ml / g) based on the amount of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate] to the reaction mixture over a period of time while maintaining the temperature at about 40 °C to about 55 °C; (b8) Cooling the reaction mixture to 20 °C ± 5.0 °C over at least 2 hours (at a rate of about 0.2 °C / min); (b9) Stirring the slurry resulting from step (b8) at 15 °C ± 5.0 °C for at least 2 hours (e.g., at least 3 hours); (b10) Filtering the slurry from step (b9); (b11) Washing the reaction vessel with water and methanol, wherein the washing liquid is cooled to 20 °C ± 5.0 °C; (b12) Transferring the washing liquid from step (b11) to a filter; (b13) Collecting the solid from the filtrate [step (b12)]; Step of drying the collected solid, optionally under vacuum, at about 45 °C to about 55 °C (e.g., 50 °C). It is a further embodiment of Embodiment G1 or G2, including

[0209] In one embodiment (Embodiment H1), the present invention is a process for preparing the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid,

[0210] [Chemical formula] (a1) Reacting methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with barium hydroxide [Ba(OH)2] in a solvent system containing an organic solvent and water to form the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, where the amount of barium hydroxide [Ba(OH)2] is about 0.5 to about 0.6 (e.g., about 0.5) molar equivalents relative to methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, (a2) Optionally, isolating the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid from step (a1), A process is provided that includes

[0211] The progress of the reaction in step (a1) can be monitored by various techniques, for example, by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). The reaction in step (a1) is carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0212] Embodiment H2 is a further embodiment of Embodiment H1, wherein the organic solvent in the solvent system in step (a1) contains an aprotic organic solvent, where the aprotic organic solvent is miscible with water. In some further embodiments, the organic solvent in the solvent system is acetone or acetonitrile. Also in some further embodiments, the volume ratio of the organic solvent and water in the solvent system is from about 4.5:1 to about 5.5:1, or from about 4.5:1 to about 5.0:1, for example, about 4.7:1, about 4.8:1, or about 5:1.

[0213] Embodiment H3 is a further embodiment of Embodiment H1 or H2, wherein the organic solvent in the solvent system in step (a1) is acetonitrile, and the volume ratio of the organic solvent and water in the solvent system is from about 4.5:1 to about 5.0:1.

[0214] Embodiment H4 is a further embodiment of any one of Embodiments H1 to H3, wherein the organic solvent in the solvent system in step (a1) is acetonitrile, the volume ratio of the organic solvent and water in the solvent system is from about 4.5:1 to about 5.0:1, and the volume amount of acetonitrile is from about 5 ml / g to about 15 ml / g based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0215] Embodiment H5 is a further embodiment of any one of Embodiments H1 to H4, in which the reaction in step (a1) is carried out under heating at a temperature of, for example, from about 40°C to about 65°C (for example, 50°C or 60°C).

[0216] Embodiment H6 is a further embodiment of Embodiment H1 or H2, in which the organic solvent in the solvent system is acetone, and the volume ratio of the organic solvent and water in the solvent system is from about 4.5:1 to about 5.5:1 (for example, 5.0:1).

[0217] Embodiment H7 is a further embodiment of Embodiment H6, in which the volume amount of acetone is from about 5 ml / g to about 15 ml / g (for example, 6 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0218] Embodiment H8 is a further embodiment of Embodiment H6 or H7, in which the reaction in step (a1) is carried out under heating at a temperature of, for example, from about 45°C to about 55°C (for example, 50°C).

[0219] The progress of the reaction in step (a1) can be monitored by various techniques, for example, by chromatographic techniques (for example, LC, TLC, or reverse-phase HPLC). At the completion of the reaction, the reaction mixture can be cooled from the elevated temperature to room temperature / ambient temperature. The hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid formed precipitates from the reaction solvent system.

[0220] Embodiment H9 is a further embodiment of any one of Embodiments H1 to H8, which includes isolating the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid by filtering the formed hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid.

[0221] Embodiment H10 is a further embodiment of any one of Embodiments H1 to H8, which includes isolating the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid by filtering the formed hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, and washing the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid obtained by filtration with the same organic solvent as that in the organic solvent system in step (a1).

[0222] Embodiment H11 is a further embodiment of any one of Embodiments H1 to H10, wherein the isolated hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid is further dried, optionally under vacuum and at an elevated temperature (e.g., from 45 °C to about 55 °C).

[0223] In one embodiment (Embodiment J1), the present invention provides a process for preparing the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, comprising:

[0224] [Chemical formula] (a1) Reacting methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with barium hydroxide [Ba(OH)2] in a solvent system comprising acetonitrile and water to form the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, wherein the amount of barium hydroxide [Ba(OH)2] is about 0.5 to about 0.6 (e.g., about 0.5) molar equivalents relative to methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate; (a2) Water, a water-immiscible organic solvent (e.g., toluene, TBME, or ethyl acetate), and an organic acid (e.g., acetic acid) are added to the reaction mixture in step (a1), and the resulting mixture is mixed to form (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, (a3) separating the organic phase / solution from the aqueous phase / solution of the mixture from step (a2); (a4) 2-Amino-2-(hydroxymethyl)-1,3-propanediol is added to the separated organic phase from step (a3), whereby 2-amino-2-(hydroxymethyl)-1,3-propanediol and (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid are reacted to form the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, where the reaction mixture is held at a holding temperature of from about 35 °C to about 55 °C (e.g., from about 40 °C to about 50 °C, or about 45 °C); (a5) adding a seed crystalline material of the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid to the reaction mixture from step (a4) to form a slurry, where the slurry is held at the holding temperature for a time longer than about 1 minute; (a6) cooling the slurry from step (a5) to a temperature of from about 20 °C to about 30 °C (e.g., 25 °C) and holding the slurry at that temperature for a time longer than about 1 minute; (a7) Optionally, heating the slurry from step (a6) to a holding temperature [from about 40 °C to about 50 °C, for example about 45 °C] and holding the slurry at the holding temperature for a time longer than about 1 minute; (a8) Optionally, cooling the slurry from step (a7) to a temperature of about 15 °C to about 25 °C (e.g., 20 °C) and holding the slurry at that temperature for a time longer than about 1 minute; (a9) Isolating the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid in the slurry from step (a6) or (a8). A process is provided that includes the above steps.

[0225] The progress of the reactions in steps (a1), (a2), and (a4) can be monitored by various techniques, such as chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). The reactions in steps (a1) and (a4) are carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0226] Embodiment J2 is a further embodiment of Embodiment J1, wherein the volume ratio of acetonitrile to water in the solvent system in step (a1) is from about 4.5:1 to about 5.5:1, or from about 4.5:1 to about 5.0:1, for example about 4.7:1, about 4.8:1, or about 5:1.

[0227] Embodiment J3 is a further embodiment of Embodiment J1 or J2, wherein the volume of acetonitrile in step (a1) is from about 5 ml / g to about 10 ml / g (e.g., about 7 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. In some further embodiments, the volume of acetonitrile in step (a1) is from about 6 ml / g to about 8 ml / g (e.g., about 7 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0228] Embodiment J4 is a further embodiment of any one of Embodiments J1 to J3, wherein the reaction in step (a1) is carried out at an elevated temperature, for example, at a temperature from about 40 °C to about 65 °C (e.g., about 50 °C or about 60 °C).

[0229] The progress of the reaction in step (a1) can be monitored by various techniques, for example, by chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). At the end of the reaction, the reaction mixture can be cooled from the elevated temperature to room temperature / ambient temperature, or to a lower temperature such as a temperature from about 20 °C to about 25 °C. The hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid precipitates from the reaction solvent system, and the reaction mixture is in the form of a slurry. This slurry is carried directly to step (a2).

[0230] Embodiment J5 is a further embodiment of any one of Embodiments J1 to J4, wherein the volume amount of water added in step (a2) is about 3 ml / g to about 4 ml / g (for example, about 3.5 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1).

[0231] Embodiment J6 is a further embodiment of any one of Embodiments J1 to J5, wherein the volume amount of the water-immiscible organic solvent (for example, toluene) added in step (a2) is about 0.8 ml / g or more based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1), and for example, it is from 0.8 ml / g to about 1.2 ml / g (for example, about 1.0 ml / g).

[0232] Embodiment J7 is a further embodiment of any one of Embodiments J1 to J6, wherein the amount of the organic acid (for example, acetic acid) added in step (a2) is about 1.0 to about 2.0 (for example, about 1.5) molar equivalents of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1). In some further embodiments, the organic acid added in step (a2) is acetic acid.

[0233] In step (a2), water and toluene are added to the reaction mixture from step (a1). An organic acid (for example, acetic acid) is added to the mixture to convert the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid to the free acid of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid. The free acid is extracted into an organic phase containing a water-immiscible organic solvent (for example, toluene).

[0234] In step (3), the organic phase containing the water-immiscible organic solvent (for example, toluene) from step (a2) is separated from the aqueous phase. The organic phase is carried directly to step (a4).

[0235] Embodiment J8 is a further embodiment of any one of embodiments J1 to J7, wherein the amount of 2-amino-2-(hydroxymethyl)-1,3-propanediol added in step (a4) is from about 1.0 to about 1.5 (for example, from about 1.1 to about 1.3, or about 1.2) molar equivalents of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1).

[0236] Embodiment J9 is a further embodiment of any one of embodiments J1 to J8, wherein the 2-amino-2-(hydroxymethyl)-1,3-propanediol added in step (a4) is in the form of an aqueous solution.

[0237] Embodiment J10 is a further embodiment of any one of embodiments J1 to J9, wherein the reaction in step (a4) is carried out with a temperature increase to about 45 °C.

[0238] Embodiment J11 is a further embodiment of any one of Embodiments J1 to J10, wherein the amount of the seed crystal is from about 0.20 to about 0.50 mol% (e.g., from about 0.25 to about 0.35 mol%, or about 0.3 mol%) of the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1) [e.g., about 0.004 g / g based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1)].

[0239] Embodiment J12 is a further embodiment of any one of Embodiments J1 to J11, which performs steps (a7) and (a8). Steps (a7) and (a8) reduce some impurities, e.g., methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0240] Embodiment J13 is a further embodiment of any one of Embodiments J1 to J12, wherein isolating the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid from the slurry from step (a6) or (a8) involves filtration, e.g., filtration under vacuum.

[0241] Embodiment J14 is a further embodiment of Embodiment J13, wherein filtration or filtration under vacuum further comprises washing the solid with methyl ethyl ketone (MEK). In some further embodiments, washing with MEK comprises 1, 2, or 3 washes. In some further embodiments, the amount of MEK used for each wash is from about 2 mL / g to about 4 mL / g (e.g., about 3 mL / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1).

[0242] Embodiment J15 is a further embodiment of any one of Embodiments J1 to J14, wherein the isolated tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid from step (a9) is further dried, for example, under vacuum at a temperature of about 40 °C or lower, about 50 °C or lower, about 60 °C or lower, or from about 45 °C to about 55 °C (e.g., 50 °C).

[0243] Embodiment J16 is a further embodiment of any one of Embodiments J1 to J15, wherein the slurry in step (a5) is held at the holding temperature for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, or longer than about 60 minutes.

[0244] Embodiment J17 is a further embodiment of any one of Embodiments J1 to J16, wherein the slurry in step (a6) is held at a temperature of about 20 °C to about 30 °C for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, or longer than about 60 minutes.

[0245] Embodiment J18 is a further embodiment of any one of Embodiments J1 to J17, in which the slurry in step (a7) is held at the holding temperature for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, or longer than about 60 minutes.

[0246] Embodiment J19 is a further embodiment of any one of Embodiments J1 to J18, in which the slurry in step (a8) is held at a temperature of about 15°C to about 25°C for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, longer than about 60 minutes, longer than about 1 hour, longer than about 2 hours, longer than about 3 hours, longer than about 4 hours, longer than about 5 hours, longer than about 6 hours, longer than about 7 hours, longer than about 8 hours, or longer than about 9 hours. In some further embodiments, the holding time in step (a8) is longer than about 4 hours, longer than about 5 hours, longer than about 6 hours, longer than about 7 hours, or longer than about 8 hours.

[0247] In one embodiment (Embodiment K1), the present invention is a process for preparing the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid,

[0248]

Chemical formula

[0249] The progress of the reactions in each of steps (a1), (a2), and (a4) can be monitored by various techniques, such as chromatographic techniques (e.g., LC, TLC, or reverse-phase HPLC). The reactions in each of steps (a1) and (a4) are carried out for a sufficient time and under such conditions to form the intended product (and to complete the reaction).

[0250] Embodiment K2 is a further embodiment of Embodiment K1, wherein the volume ratio of acetone to water in the solvent system in step (a1) is from about 4.5:1 to about 5.5:1, for example about 5:1.

[0251] Embodiment K3 is a further embodiment of Embodiment K1 or K2, wherein the volume amount of acetone in step (a1) is from about 5 ml / g to about 8 ml / g (e.g., about 6 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. In some further embodiments, the volume amount of acetonitrile in step (a1) is from about 5 ml / g to about 7 ml / g (e.g., about 6 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.

[0252] Embodiment K4 is a further embodiment of any one of Embodiments K1 to K3, wherein the reaction in step (a1) is carried out at an elevated temperature, for example, at a temperature from about 40°C to about 60°C (e.g., from about 45°C to about 55°C, or about 50°C).

[0253] Embodiment K5 is a further embodiment of any one of Embodiments K1 - K4, wherein the volume amount of water added in step (a2) is from about 2.5 ml / g to about 4 ml / g (e.g., about 3.0 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1).

[0254] Embodiment K6 is a further embodiment of any one of Embodiments K1 - K5, wherein the volume amount of the water-immiscible organic solvent (e.g., toluene, TBME, or ethyl acetate) added in step (a2) is about 2.0 ml / g or more, for example, from 2.0 ml / g to about 4.0 ml / g, or from 3.0 ml / g to about 4.0 ml / g (e.g., about 3.5 ml / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1). In some further embodiments, the water-immiscible organic solvent is toluene.

[0255] Embodiment K7 is a further embodiment of any one of Embodiments K1 - K6, wherein the amount of the organic acid (e.g., acetic acid) added in step (a2) is from about 1.0 to about 2.0 (e.g., about 1.5) molar equivalents of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1). In some further embodiments, the organic acid added in step (a2) is acetic acid.

[0256] The progress of the reaction in step (a1) can be monitored by various techniques, for example, by chromatographic techniques (such as LC, TLC, or reverse-phase HPLC). At the end of the reaction, the reaction mixture can be cooled from an elevated temperature to room temperature / ambient temperature, or to a lower temperature such as from about 20 °C to about 30 °C (for example, from about 22 °C to about 28 °C, or from about 23 °C to about 27 °C, or about 25 °C). The hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid formed precipitates from the reaction solvent system, and the reaction mixture is in the form of a slurry. This slurry is carried directly to step (a2).

[0257] In step (a2) of some embodiments, water and toluene are added to the reaction mixture from step (a1). An organic acid (such as acetic acid) is added to the mixture to convert the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid to the free acid of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid. After the addition of the organic acid, the reaction mixture in step (a2) is stirred for a sufficient time and under conditions (such as at a temperature from about 20 °C to about 30 °C, for example, at 25 °C) to complete the reaction. The free acid formed in step (a2) is then extracted into an organic phase containing a water-immiscible organic solvent (such as toluene) in step (a3).

[0258] In step (a3), the organic phase containing the water-immiscible organic solvent (such as toluene) from step (a2) is separated from the aqueous phase. After separation, methanol may be added to the organic phase, and the resulting solution is carried directly to step (a4).

[0259] Embodiment K8 is a further embodiment of any one of Embodiments K1 to K7, wherein the amount of methanol used in step (a3), if present, is from about 0.5 mL / g to 1.0 mL / g (e.g., 0.75 mL / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1).

[0260] Embodiment K9 is a further embodiment of any one of Embodiments K1 to K8, wherein the amount of 2-amino-2-(hydroxymethyl)-1,3-propanediol added in step (a4) is from about 1.0 to about 1.5 (e.g., from about 1.1 to about 1.3, or about 1.2) molar equivalents of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1).

[0261] Embodiment K10 is a further embodiment of any one of Embodiments K1 to K9, wherein the 2-amino-2-(hydroxymethyl)-1,3-propanediol added in step (a4) is in the form of an aqueous solution.

[0262] Embodiment K11 is a further embodiment of any one of Embodiments K1 to K10, wherein the reaction in step (a4) is carried out with a temperature increase of about 45 °C.

[0263] Embodiment K12 is a further embodiment of any one of embodiments K1 to K11, wherein the amount of the seed crystal is from about 0.20 to about 0.50 mol% (e.g., from about 0.25 to about 0.35 mol%, or about 0.3 mol%) of the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1) [e.g., about 0.004 g / g based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1)]. In some further embodiments, after seeding, the slurry is stirred for a time longer than about 1 hour.

[0264] Embodiment K13 is a further embodiment of any one of embodiments K1 to K12, wherein isolating the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid in step (a7) involves filtration, e.g., filtration under vacuum.

[0265] Embodiment K14 is a further embodiment of Embodiment K13, wherein the filtration or filtration under vacuum further comprises washing the solid with acetone. In some further embodiments, washing with acetone comprises 1, 2, or 3 washes (e.g., 2 washes). In some further embodiments, the amount of acetone used for each wash is about 2 mL / g to about 4 mL / g (e.g., about 3 mL / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1).

[0266] Embodiment K15 is a further embodiment of any one of Embodiments K1 to K14, wherein the isolated tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid from step (a7) is further dried, for example, under vacuum at a temperature of about 40 °C or lower, about 50 °C or lower, about 60 °C or lower, or from about 45 °C to about 55 °C (e.g., 50 °C).

[0267] Embodiment K16 is a further embodiment of any one of Embodiments K1 to K15, wherein the slurry in step (a5) is held at the holding temperature for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, or longer than about 60 minutes.

[0268] Embodiment K17 is a further embodiment of any one of Embodiments K1 to K16, wherein the slurry in step (a6) is held at a temperature of from about 15°C to about 20°C for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, longer than about 60 minutes, longer than about 1 hour, longer than about 2 hours, longer than about 3 hours, longer than about 4 hours, longer than about 5 hours, longer than about 6 hours, longer than about 7 hours, longer than about 8 hours, or longer than about 9 hours. In some further embodiments, the holding time in step (a8) is longer than about 4 hours, longer than about 5 hours, longer than about 6 hours, longer than about 7 hours, or longer than about 8 hours.

[0269] In one embodiment (Embodiment L1), the present invention is a method for preparing Form 1 of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, comprising: (a) suspending the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid in a solvent system, wherein the solvent system consists of dimethyl sulfoxide (DMSO) and water, the volume ratio of DMSO:water is from about 10:1 to about 6:1 (e.g., from about 9:1 to about 7:1, or about 8:1), and the ratio of water (volume) to the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (weight) is from about 0.8 mL / g to about 1.2 mL / g (e.g., 1.0 mL / g) at a temperature from about 20°C to about 35°C (e.g., from about 20°C to about 30°C); (b) Heating the suspension in step (a) to a temperature rise from about 60 °C to about 70 °C (e.g., about 65 °C) to form a solution, and then mixing the solution for a time longer than about 1 minute during the temperature rise; (c) Slowly adding water to the solution from step (b) while maintaining the reaction mixture as a solution, where the amount of water added is approximately the same as the water used in step (a), and then holding the resulting solution for a time longer than about 1 minute during the temperature rise; (d) Seeding the solution with the crystalline form 1 material of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid while maintaining the temperature at the temperature rise, where the amount of the seed crystal crystallinity is at least about 0.5 wt% (e.g., about 0.5 wt% or about 1.0 wt%) of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid used in step (a), and then holding the resulting mixture for a time longer than about 1 minute during the temperature rise; (e) Optionally, slowly cooling the temperature of the mixture from step (d) to about 30 °C (e.g., at a rate of about 0.2 °C / min), and then holding the mixture at that temperature for a time longer than about 1 minute; (f) Optionally, slowly heating the mixture from step (e) to about 40 °C to about 50 °C (e.g., at a rate of about 0.5 °C / min), and then holding the mixture at that temperature for a time longer than about 1 minute; (g) Slowly cooling the temperature of the mixture from step (d) or step (f) (when steps (e) and (f) are carried out) to about 15 °C (e.g., at a rate of about 0.2 °C / min), and then holding the mixture at that temperature for a time longer than about 1 minute; (h) Isolating a solid from the mixture resulting from step (g) to obtain Form I of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid and providing a method comprising the same.

[0270] Embodiment L1 relates to a recrystallization process in which (1) a solvent system consisting of dimethyl sulfoxide (DMSO) and water is used to dissolve the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid at an elevated temperature, (2) while maintaining the resulting mixture as a solution, water (a poor solvent) is added to the solution at an elevated temperature, (3) the solution is seeded with crystalline Form I material at an elevated temperature, (4) the resulting mixture is cooled to a low temperature (e.g., about 15 °C), (5) optionally, the mixture is heated and then cooled, and (6) crystalline Form I is isolated from the cooled mixture.

[0271] Embodiment L2 is a further embodiment of Embodiment L1, wherein the volume ratio of DMSO:water in step (a) is from about 9:1 to about 7:1.

[0272] Embodiment L3 is a further embodiment of Embodiment L1 or L2, wherein the volume ratio of DMSO:water in step (a) is about 8:1.

[0273] Embodiment L4 is a further embodiment of any one of Embodiments L1 to L3, wherein the ratio of water (volume) to the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (weight) in step (a) is about 1.0 mL / g.

[0274] Embodiment L5 is a further embodiment of any one of Embodiments L1 to L4, where in step (b), the suspension is heated at an elevated temperature (e.g., about 65°C) to form a solution. In some further embodiments, the solution in step (b) is mixed at an elevated temperature for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, or longer than about 30 minutes.

[0275] Embodiment L6 is a further embodiment of any one of Embodiments L1 to L5, where in step (c), water is slowly added so that the mixture is maintained as a solution and the temperature of the solution is maintained without deviating from an elevated temperature above 5.0°C. In some further embodiments, the holding time in step (c) is longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, or longer than about 30 minutes.

[0276] Embodiment L7 is a further embodiment of any one of Embodiments L1 to L6, where the seeding in step (d) is carried out so that the temperature of the mixture is maintained without deviating from an elevated temperature above 5.0°C, and at least some of the seeding material remains solid after the seeding is completed and some solids are present after the holding period in step (d). In some further embodiments, the holding time in step (d) is longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, or longer than about 60 minutes. In some further embodiments, the holding time in step (e) is longer than about 1 hour.

[0277] Embodiment L8 is a further embodiment of any one of Embodiments L1 to L7, where steps (e) and (f) are carried out.

[0278] Embodiment L9 is a further embodiment of any one of Embodiments L1 to L8, wherein the cooling in step (e) is performed at a rate of less than about 1 °C / min, less than about 0.8 °C / min, less than about 0.5 °C / min, less than about 0.3 °C / min (e.g., about 0.2 °C / min). In some further embodiments, the cooling in step (e) is performed at a rate of about 0.2 °C / min.

[0279] Embodiment L10 is a further embodiment of any one of Embodiments L1 to L9, wherein the holding time in step (e) is longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, or longer than about 1 hour. In some further embodiments, the holding time in step (e) is longer than about 1 hour.

[0280] Embodiment L11 is a further embodiment of any one of Embodiments L1 to L10, wherein the heating in step (f) is performed at a rate of less than about 1 °C / min, less than about 0.8 °C / min, less than about 0.7 °C / min, less than about 0.6 °C / min, less than about 0.5 °C / min, less than about 0.3 °C / min. In some further embodiments, the heating in step (f) is performed at a rate of about 0.5 °C / min.

[0281] Embodiment L12 is a further embodiment of any one of Embodiments L1 to L11, wherein the holding time in step (f) is longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, or longer than about 30 minutes. In some further embodiments, the holding time in step (f) is about 30 minutes.

[0282] Embodiment L13 is a further embodiment of any one of Embodiments L1 to L12, wherein the cooling in step (g) is performed at a rate of less than about 1 °C / min, less than about 0.8 °C / min, less than about 0.5 °C / min, or less than about 0.3 °C / min (e.g., about 0.2 °C / min). In some further embodiments, the cooling in step (g) is performed at a rate of about 0.2 °C / min.

[0283] Embodiment L14 is a further embodiment of any one of Embodiments L1 to L13, wherein the holding time in step (g) is longer than about 1 hour, longer than about 2 hours, longer than about 3 hours, longer than about 4 hours, longer than about 5 hours, longer than about 6 hours, longer than about 7 hours, longer than about 8 hours, or longer than about 9 hours. In some further embodiments, the holding time in step (g) is longer than about 8 hours.

[0284] Embodiment L15 is a further embodiment of any one of Embodiments L1 to L14, wherein isolating the solid in step (f) includes filtration, for example, filtration under vacuum.

[0285] Embodiment L16 is a further embodiment of Embodiment L15, wherein filtration or filtration under vacuum further includes washing the solid with methyl ethyl ketone (MEK). In some further embodiments, washing with MEK includes 1, 2, or 3 washes (for example, 2 washes). Also, in some further embodiments, the amount of MEK used for each wash is about 2 mL / g to about 4 mL / g (for example, about 3 mL / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1). Additionally, in some further embodiments, the MEK used for each wash is cooled to about 15 °C before washing.

[0286] Embodiment L16 is a further embodiment of any one of Embodiments L1 to L15, wherein the isolated solid from step (h) is further dried, for example, under vacuum at a temperature of about 40 °C or lower, about 50 °C or lower, about 60 °C or lower, or from about 50 °C to about 60 °C (for example, 55 °C).

[0287] In one embodiment (Embodiment M1), the present invention is a method for preparing Form 1 of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, (a) suspending the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid in a solvent system, where the solvent system consists of tetrahydrofuran (THF) and water, the volume ratio of THF:water is from about 1:1 to about 4:1 (e.g., from about 2.5:1 to about 3.5:1, from about 2.8:1 to about 3.2:1, from about 2.9:1 to about 3.1:1, or about 3.0:1), and the ratio of water (volume) to the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (weight) is from about 1.1 mL / g to about 3.8 mL / g (e.g., from about 1.3 mL / g to about 1.5 mL / g, or about 1.39 mL / g) at a temperature from about 20 °C to about 35 °C (e.g., from about 20 °C to about 30 °C, about 22 °C); (b) heating the suspension in step (a) to a high temperature from about 49 °C to about 59 °C (e.g., about 55 °C) to form a solution, cooling the temperature to a holding temperature from about 47 °C to about 51 °C (e.g., about 49 °C), during which the mixture remains a solution, and optionally, mixing the solution at the holding temperature for a time longer than about 1 minute; (c) To the solution from step (b), crystalline form 1 material of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid is seeded while maintaining the temperature at the holding temperature, where the amount of seed crystal crystallinity is about 0.5% by weight or more (e.g., about 0.5% by weight or about 1.0% by weight) of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid used in step (a), and then the resulting mixture is held at the holding temperature for a time longer than about 1 minute, (d) The temperature of the mixture from step (c) is slowly cooled to an intermediate temperature of about 35 °C, and then the mixture is held at the intermediate temperature for a time longer than about 1 minute, (e) A water-miscible organic solvent (e.g., acetonitrile, isopropanol, or acetone) is slowly added to the mixture from step (d) while maintaining the temperature of the mixture at the intermediate temperature, and then the mixture is held at the intermediate temperature for a time longer than about 1 minute, (f) The temperature of the mixture from step (e) is slowly cooled to a low temperature of about 10 °C, and then the mixture is held at the low temperature for a time longer than about 1 minute, (g) Taking a sample of the mixture (slurry) and determining the particle size of the solid in the mixture, (h) Performing high-shear wet grinding until the D90 of the particle size of the solid in the mixture is less than about 150 μM, (i) Isolating the solid from the resulting mixture from step (f) to obtain form I of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid A method is provided that includes the above steps.

[0288] Embodiment M1 relates to a recrystallization process in which a tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid is dissolved at a high temperature (e.g., 55 °C) using a solvent system consisting of THF and water, then the solution is cooled to a holding temperature (e.g., 49 °C), (2) the solution is seeded with crystalline form I material at the holding temperature, (3) the resulting mixture is cooled to an intermediate temperature (e.g., 35 °C), acetonitrile is added at the intermediate temperature, (4) the resulting mixture is cooled to a low temperature (e.g., about 10 °C), and (5) crystalline form I is isolated from the cooled mixture.

[0289] Embodiment M2 is a further embodiment of Embodiment M1, wherein the volume ratio of THF:water in step (a) is from about 2.9:1 to about 3.1:1.

[0290] Embodiment M3 is a further embodiment of Embodiment M1 or M2, wherein the volume ratio of THF:water in step (a) is about 3.0:1.

[0291] Embodiment M4 is a further embodiment of any one of Embodiments M1 to M3, wherein the ratio of water (volume) to the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (weight) in step (a) is about 1.39 mL / g.

[0292] Embodiment M5 is a further embodiment of any one of Embodiments M1 to M4, in which first in step (b), the suspension is heated to a high temperature from about 53°C to about 59°C (for example, about 55°C) to form a solution, and then the temperature of the solution is cooled to a holding temperature from about 48°C to about 50°C, during which the mixture remains in solution. In some further embodiments, first in step (b), the suspension is heated at a high temperature from about 53°C to about 57°C (for example, about 55°C) to form a solution, and then the temperature of the solution is cooled to a holding temperature of about 49°C. In some further embodiments, the solution in step (b) is mixed at the holding temperature for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, or longer than about 1 hour.

[0293] Embodiment M6 is a further embodiment of any one of Embodiments M1 to M5, in which the seeding in step (c) is carried out such that the temperature of the mixture is maintained without deviating from a holding temperature above 1.0°C or above 2.0°C, and at least some of the seeding material remains solid after the seeding is completed and some solids are present after the holding period in step (c). In some further embodiments, after seeding, the mixture in step (c) is mixed at the holding temperature for a time longer than about 1 minute, longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, longer than about 60 minutes, or longer than about 75 minutes.

[0294] Embodiment M7 is a further embodiment of any one of Embodiments M1 to M6, in which the cooling in step (d) is carried out at a rate of less than about 0.5 °C / min, less than about 0.4 °C / min, less than about 0.3 °C / min, less than about 0.2 °C / min (for example, from about 0.1 °C / min to about 0.2 °C / min). In some further embodiments, the cooling in step (d) is carried out at a rate of from about 0.1 °C / min to about 0.2 °C / min. Also, in some further embodiments, after the mixture in step (d) is cooled to the intermediate temperature, the mixture is held at the intermediate temperature for a time longer than about 5 minutes, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, longer than about 60 minutes, or longer than about 75 minutes.

[0295] Embodiment M8 is a further embodiment of any one of Embodiments M1 to M7, in which in step (e), a water-miscible organic solvent (for example, acetonitrile) is slowly added so that the temperature of the mixture is maintained without deviating from an intermediate temperature above 1.0 °C, above 2.0 °C, or above 3.0 °C. In some embodiments, the water-miscible organic solvent is acetonitrile. In some further embodiments, the addition of acetonitrile is carried out over a time exceeding 1 hour, exceeding 1.5 hours, or exceeding 2 hours.

[0296] Embodiment M9 is a further embodiment of any one of Embodiments M1 to M8, in which after the addition of acetonitrile in step (e) is completed, the resulting mixture is held at the intermediate temperature for a time longer than about 1 minute, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, longer than about 60 minutes, or longer than about 75 minutes.

[0297] Embodiment M10 is a further embodiment of any one of embodiments M1 to M9, in which the cooling in step (f) is carried out at a rate of less than about 0.5 °C / min, less than about 0.4 °C / min, less than about 0.3 °C / min, or less than about 0.2 °C / min (for example, about 0.2 °C / min). In some further embodiments, the cooling in step (f) is carried out at a rate of about 0.2 °C / min.

[0298] Embodiment M11 is a further embodiment of any one of embodiments M1 to M10, in which the holding time in step (f) is longer than about 1 minute, longer than about 10 minutes, longer than about 15 minutes, longer than about 20 minutes, longer than about 30 minutes, longer than about 45 minutes, longer than about 1 hour, longer than about 2 hours, longer than about 3 hours, longer than about 4 hours, longer than about 5 hours, longer than about 6 hours, longer than about 7 hours, longer than about 8 hours, or longer than about 9 hours. In some further embodiments, the holding time in step (f) is longer than about 5 hours, about 6 hours, about 7 hours, or about 8 hours.

[0299] Embodiment M12 is a further embodiment of any one of embodiments M1 to M11, in which after performing high-shear wet grinding on a sample of the mixture in step (h) to determine the particle size of the solid in the mixture, if the D90 of the particle size of the solid in the mixture is greater than about 125 μM, high-shear wet grinding is performed again until the D90 value is less than about 125 μM.

[0300] Embodiment M13 is a further embodiment of any one of embodiments M1 to M12, in which isolating the solid in step (i) includes filtration, for example, filtration under vacuum.

[0301] Embodiment M14 is a further embodiment of Embodiment M13, wherein the filtration or filtration under vacuum further comprises washing the solid with methyl ethyl ketone (MEK). In some further embodiments, washing with MEK comprises 1, 2, or 3 washes (e.g., 2 washes). In some further embodiments, the amount of MEK used for each wash is from about 2 mL / g to about 2.5 mL / g (e.g., about 2.2 mL / g) based on the amount of methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate used in step (a1). In yet further embodiments, the MEK used for each wash may be at room temperature cooled to about 10 °C before washing.

[0302] Embodiment M15 is a further embodiment of any one of Embodiments M1 to M14, wherein the isolated solid from step (i) is further dried, for example, under vacuum at a temperature of about 40 °C or lower, about 50 °C or lower, about 60 °C or lower, or from about 50 °C to about 60 °C (e.g., 55 °C).

[0303] Preparation C111, its tris salt, the solid form of the tris salt of C111, and certain intermediates can be prepared by the general and specific methods described hereinafter, which are linked to the common general knowledge of those skilled in the art of synthetic organic chemistry and / or solid forms of pharmaceutical compounds. Such common general knowledge can be found in standard reference books such as Comprehensive Organic Chemistry, edited by Barton and Ollis, Elsevier; Comprehensive Organic Transformations: A Guide to Functional Group Preparations, Larock, John Wiley and Sons; and Compendium of Organic Synthetic Methods, Volumes I - XII (published by Wiley - Interscience). The starting materials used herein can be commercially available or prepared by routine methods known in the art.

[0304] In the preparation of the compounds, salts, and solid forms (e.g., crystalline and amorphous) of the present invention, it should be noted that some of the preparation methods described herein may require the protection of remote functional groups (e.g., primary amines, secondary amines, carboxyls in the precursors). The need for such protection will vary depending on the nature of the remote functional group and the conditions of the preparation method. The need for such protection can be easily determined by those skilled in the art. The use of such protection / deprotection methods is also within the scope of the skills possessed by those skilled in the art. For an overview of protecting groups and their use, see T.W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0305] For example, a particular compound contains a primary amine or carboxylic acid functional group that, if left unprotected, can interfere with reactions at other sites of the molecule. Thus, such functional groups may be protected by appropriate protecting groups that can be removed in a subsequent step. Protecting groups suitable for amine and carboxylic acid protection include those commonly used in peptide synthesis (for amines, N-t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), and 9-fluorenylmethyloxycarbonyl (Fmoc), and for carboxylic acids, lower alkyl or benzyl esters, etc.), which are generally chemically non-reactive under the described reaction conditions and can typically be removed without chemically altering other functional groups in the compound.

[0306] The following description is intended to provide an overview of the methodologies used in the preparation of the compounds and solid forms of the present invention. Some of the compounds of the present invention may contain single or multiple chiral centers with stereochemical designations (R) or (S). It will be apparent to those skilled in the art that all synthetic transformations can be carried out in a similar manner regardless of whether the material is enantiomerically enriched or racemic. Further, resolution to the desired optically active material can occur at any desired point in the sequence using well-known methods such as those described herein and in the chemical literature. For example, intermediates and final products may be separated using chiral chromatographic methods. Alternatively, chiral salts may be utilized to isolate enantiomerically enriched intermediates and final compounds.

Examples

[0307] The following illustrates the synthesis of non-limiting compounds of the present invention (including their solid forms).

[0308] The experiments were generally carried out under an inert atmosphere (nitrogen or argon), particularly in cases where reagents or intermediates sensitive to oxygen or moisture were used. Commercially available solvents and reagents were generally used without further purification. Where appropriate, anhydrous solvents, generally AcroSeal® products from Acros Organics, Aldrich® Sure / Seal™ products from Sigma - Aldrich, or DriSolv® products from EMD Chemicals were used. In other cases, commercially available solvents were passed through columns packed with 4 Å molecular sieves until they reached the following QC criteria for water: a) less than 100 ppm for dichloromethane, toluene, N,N - dimethylformamide, and tetrahydrofuran; b) less than 180 ppm for methanol, ethanol, 1,4 - dioxane, and diisopropylamine. For highly sensitive reactions, the solvents were further treated with metallic sodium, calcium hydride, or molecular sieves and distilled immediately before use. The products were generally dried under vacuum and then carried over to further reactions or submitted for biological testing. Mass spectrometry data was reported by either liquid chromatography - mass spectrometry (LCMS), atmospheric pressure chemical ionization (APCI), or gas chromatography - mass spectrometry (GCMS) equipment. The symbol ◆ indicates that the chlorine isotope pattern was observed in the mass spectrum.

[0309] Chiral separation was used to separate enantiomers or diastereomers of some intermediates during the preparation of the compounds of the present invention.

[0310] Following reactions proceeding via detectable intermediates, LCMS was generally performed and allowed to proceed to complete conversion before the addition of subsequent reagents. In syntheses referring to procedures in other examples or methods, the reaction conditions (reaction time and temperature) may vary. Generally, following the reaction, thin - layer chromatography or mass spectrometry was performed and, where appropriate, subjected to work - up. Purification may vary between experiments and generally, the solvents and solvent ratios used for the eluent / gradient were appropriate for the R for selected to provide a retention time. All starting materials in these preparations and examples are either commercially available or can be prepared by methods known in the art or as described herein.

[0311] (Example 1) Preparation of 3-Fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt (anhydrous)

[0312] [Chemical formula] Step 1. Preparation of C103 [tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate] The clean and dry reactor was evacuated and filled with nitrogen to atmospheric pressure until the oxygen content was 1.0% or less. 1,4-Dioxane (594 kg, 8.3 kg / kg) was charged into the reactor at 15 to 25 °C. While maintaining the temperature at 15 to 25 °C, C101 [72 kg, 1.0 equivalent (limiting reagent), tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate] was added to the mixture, followed by C102 (38.5 kg, 1.05 equivalents, 3-fluoro-4-(hydroxymethyl)benzonitrile), and the mixture was stirred for 10 to 20 minutes. Cesium carbonate (132.5 kg, 1.8 kg / kg) was added to the mixture under nitrogen protection, followed by tris(dibenzylideneacetone)dipalladium (2.4 kg, 0.01 equivalent, Pd2dba3) and 2-(di-tert-butylphosphino)biphenyl (JohnPhos 1.6 kg, 0.02 equivalent). The mixture was heated to 83 to 90 °C under nitrogen protection. After 8 to 16 hours, at the completion of the reaction, the reaction mixture was cooled to 15 to 25 °C. The mixture was filtered through celite (25 kg, 0.2 kg / kg). The filter cake was washed with MTBE (3 × 270 kg, 3 × 3.75 kg / kg, methyl tert-butyl ether). To the combined filtrate at 10 to 35 °C, a silicon-based metal scavenger (72 kg, 1 kg / kg) was added. The mixture was stirred at 40 to 50 °C for 12 to 18 hours. After 12 - 18 hours, at the completion of the reaction (confirmed by LC), the mixture was cooled to 20 to 30 °C, filtered, and the cake was washed with MTBE (324 kg, 4.5 kg / kg). The filtrate was transferred through an in-line filter and concentrated under reduced pressure (P ≤ -0.08 MPa) at T ≤ 50 °C to a final volume of 2.8 to 3.5 L / kg. Absolute ethanol (182 kg, 2.5 kg / kg) was added, and the mixture was concentrated under reduced pressure at T ≤ 50 °C to a final volume of 2.8 to 3.5 L / kg. This process was repeated 3 more times to obtain a concentrated mixture containing C103.

[0313] Step 2. Preparation of 3-Fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile Bis(4-methylbenzenesulfonate) Salt (C104 Bis(4-methylbenzenesulfonate)) Absolute ethanol (193 kg, 2.7 kg / kg) was added to the concentrated mixture containing C103 from Step 1. In a separate vessel, a solution of p-toluenesulfonic acid (118 kg, 2.55 equivalents) in absolute ethanol (144 kg, 2.0 kg / kg) was prepared at 15 to 25 °C and complete dissolution was confirmed. The prepared p-toluenesulfonic acid / ethanol solution was added to the C103-containing ethanol mixture. The mixture was heated to 55 to 65 °C under nitrogen protection. After 2 to 4 hours, the mixture was sampled for HPLC analysis. While maintaining the temperature at 55 to 65 °C, ethyl acetate (580 kg, 8 kg / kg) was added to the mixture. The mixture was cooled to 0 to 5 °C to cause crystallization. The mixture was slurried at 0 to 5 °C for 2 to 5 hours and then filtered. The filter cake was washed with ethyl acetate (2 × 90 kg, 1.25 kg / kg). The solid was dried at 30 to 40 °C for 8 to 16 hours. After cooling the solid to 15 to 30 °C, it was collected (weight 135 kg). Karl Fischer titration was performed to determine that the collected solid was anhydrous. 1 H NMR (600 MHz, DMSO-d6) δ: 8.53 (br s, 1H), 8.26 (br s, 1H), 7.89 (d, 1H), 7.67 - 7.78 (m, 3H), 7.48 (d, 4H), 7.11 (d, 4H), 6.90 (d, 1H), 6.79 (d, 1H), 5.48 (s, 2H), 3.35 (d, 2H), 2.96 - 3.09 (m, 2H), 2.79 - 2.96 (m, 1H), 2.29 (s, 6H), 1.93 - 2.03 (m, 2H), 1.77 - 1.90 (m, 2H).

[0314] The relative amounts of each reagent used in this example refer to the limiting reagent, which is C101.

[0315] (Example 2) Alternative Preparation of 3-Fluoro-4-(((6-(Piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile Bis(4-methylbenzenesulfonate) Salt (Anhydrous)

[0316] [Chemical Formula] Step 1. Preparation of C103 [tert-Butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate] Into a clean reactor vessel, C101 (25 g, 1.0 equivalent, tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate, limiting reagent), C102 (13.5 g, 1.05 equivalents, 3-fluoro-4-(hydroxymethyl)benzonitrile), and tribasic potassium phosphate (31.1 g, 1.7 equivalents) were charged. Anisole (200 ml, 8 ml / g) was charged into the reactor at once, and the reactor was stirred at 25 °C under nitrogen sweep to degas the system (target <0.15% w / w O2). In a separate vessel, palladium(II) acetate (0.05 g, 0.0025 equivalent) was dissolved in anisole (1.0 ml, 0.04 ml / g) at 20 °C to obtain a clear pale orange / brown solution. In a separate vessel, X-Phos (0.1 g, 0.0025 equivalent) was dissolved in anisole (1.0 ml, 0.04 ml / g) to obtain a clear solution. The solutions of palladium acetate and X-Phos were charged into the reactor vessel at once via syringe. The resulting suspension was stirred at 100 °C for 20 h, then sampled to confirm the completion of the reaction (LC). At the completion of the reaction, the reaction mixture was cooled to 20 - 25 °C, ethanol (50 ml, 2 mL / g) was added to the clear orange solution at 20 - 25 °C, and then filtered through Arbocel® (approx. 6.25 g, 0.25 g / g). The Arbocel® filter cake was washed with ethyl acetate (100 ml, 4 ml / g).

[0317] Step 2. Preparation of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt (C104 bis(4-methylbenzenesulfonate)) The filtrate obtained from Step 1 was charged into a clean reactor vessel. p-Toluenesulfonic acid monohydrate (36 g, 2.2 equivalents) was added all at once at 20 - 25 °C, and the suspension was stirred until completely dissolved. The reaction mixture was stirred at 40 °C for 20 hours (overnight), and then a sample was taken to check for completion of the reaction. At the completion of the reaction, the pale yellow suspension was cooled to 0 °C and stirred for at least 1 hour. The suspension was filtered under vacuum with anisole (100 ml, 4 ml / g), and used in two portions (50 ml, 2 ml / g per portion) as the cake wash liquid. The filter cake was further washed with ethyl acetate (50 ml, 2 ml / g) and then transferred to a vacuum oven. The product was dried under vacuum at 50 °C for 16 hours (overnight) and then additionally dried at 60 °C for 16 hours (overnight). The crude C104 bis(4-methylbenzenesulfonate) salt was isolated as a white to off-white solid (48.3 g, 87% yield). Karl Fischer titration was performed to determine that the white to off-white solid was anhydrous.

[0318] The relative amounts of each reagent used in this example refer to the limiting reagent which is C101.

[0319] Powder X-ray diffraction (PXRD) method (used here for all PXRD data described in this patent). The powder X-ray diffraction pattern was generated using a Bruker AXS D8 Endeavor diffractometer equipped with a Cu radiation source. The tube voltage and amperage were set to 40 kV and 40 mA, respectively. The motorized divergence slit was set to 11 mm of continuous illumination. The diffracted radiation was detected using a LYNXEYE XE-T energy-dispersive X-ray detector with a position-sensitive detector (PSD) aperture set to 4.00°. Data were collected on a theta-theta goniometer using a step width of 0.019° 2θ and a time per step of 0.2 seconds at a Cu wavelength from 2.0 to 55.0 degrees two-theta (°2θ). Samples were prepared for analysis by placing them in a silicon low-background small sample holder and rotated at 15 rpm during data collection. The data were analyzed using DIFFRAC.EVA V5.0 software. The peak list was prepared using reflections with a relative intensity of more than 5% of the most intense band in each respective diffraction pattern. A typical error of ±0.2° 2θ at the peak position (USP-941) applies to this data. Minor errors associated with this measurement can arise for various reasons including (a) sample preparation (e.g., sample height), (b) instrument characteristics, (c) instrument calibration, (d) operator input (e.g., in determining peak locations), and (e) material properties (e.g., preferred orientation and transparency effects).

[0320] A sample of the anhydrous bistosylate of C104 (as prepared by the procedure of either Example 1 or 2) was used to generate a powder X-ray diffraction pattern (designated as Form 1 herein). The PXRD pattern for Form 1 of the bistosylate of C104 is provided in Figure 1, and the corresponding peak list is provided in Table E2 (along with the peaks whose relative intensity is 5% or more).

[0321] [Table 1]

[0322] (Example 3) Preparation of 3-Fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile Bis(4-methylbenzenesulfonate) Salt (Monohydrate)

[0323] [Chemical Formula] Step 1. Preparation of C103 [tert-Butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate] Into a clean reactor vessel, C101 [25 g, 1.0 equivalent, limiting reagent, tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate], C102 (13.5 g, 1.05 equivalents, 3-fluoro-4-(hydroxymethyl)benzonitrile) and tribasic potassium phosphate (31.1 g, 1.7 equivalents) were charged. Anisole (200 ml, 8 ml / g) was charged into the reactor at once, and the reactor was stirred at 25 °C under nitrogen sweeping to degas the system (target <0.15% w / w O2). In a separate vessel, palladium(II) acetate (0.05 g, 0.0025 equivalent) was dissolved in anisole (5.0 ml, 0.2 ml / g) at 20 °C to obtain a clear pale orange / brown solution. In a separate vessel, X-Phos (0.1 g, 0.0025 equivalent) was dissolved in anisole (5.0 ml, 0.2 ml / g) to obtain a clear solution. The solutions of palladium acetate and X-Phos were charged into the reactor vessel at once via a syringe. The resulting suspension was stirred at 100 °C for 16 hours, and then a sample was taken to confirm the completion of the reaction (LC). Upon completion of the reaction, the reaction mixture was cooled to 20 - 25 °C, and water (75 ml, 3 ml / g), ethyl acetate (100 ml, 4 ml / g) and ethanol (50 ml, 2 ml / g) were added to the reaction mixture. The layers were allowed to settle and the phases were separated.

[0324] Step 2. Preparation of 3-Fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile Bis(4-methylbenzenesulfonate) Salt (C104 Bis(4-methylbenzenesulfonate)) p-Toluenesulfonic acid monohydrate (40.7 g, 2.5 eq) was added all at once to the organic phase (from Step 1) at 20 - 25 °C, and the reaction mixture was heated to 40 °C. After 20 h, a sample of the slurry was taken to confirm completion of the reaction. At completion of the reaction, the suspension was cooled to 5 °C and stirred for at least 1 h. The suspension was filtered under vacuum, and the filter cake was washed twice with ethyl acetate (2 x 50 ml, 2 x 2 ml / g) and then transferred to a vacuum oven. The product was dried under vacuum at 50 °C for 16 h (overnight) and then additionally dried at 60 °C for 16 h (overnight). The crude C104 bis(4-methylbenzenesulfonate) salt was isolated as a white - off - white solid (46.4 g, 84% yield). Karl Fischer titration was performed to determine that the white - off - white solid was the monohydrate.

[0325] The relative amounts of each reagent used in this example refer to the limiting reagent which is C101.

[0326] [Table 2]

[0327] A sample of the monohydrate bistosylate of C104 (as prepared by the procedure described in Example 3) was used to generate a powder X - ray diffraction pattern (designated as Form 2 herein). The PXRD pattern for Form 2 (monohydrate) of the bistosylate of C104 is provided in Figure 2, and the corresponding peak list is provided in Table E3 (along with the peaks having a relative intensity of 5% or more).

[0328] [Table 3]

[0329] (Example 4) Further alternative preparation of 3 - fluoro - 4 - ((((6 - (piperidin - 4 - yl)pyridin - 2 - yl)oxy)methyl)benzonitrile bis(4 - methylbenzenesulfonate) salt (anhydrous)

[0330]

Chem.

[0331] Step 2. Preparation of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt [C104 bis(4-methylbenzenesulfonate)] The oil obtained from Step 1 was redissolved in 1,4-dioxane (5 ml, 1.67 ml / g) and MTBE (2.5 ml, 0.83 ml / g), p-toluenesulfonic acid monohydrate (4.1 g, 2.1 equivalents) was added, and then the resulting mixture was heated to 50 °C. Additional 1,4-dioxane (10 ml, 3.33 ml / g) was added to collect the slurry. The solid was isolated by filtration, washed with 1:1 1,4-dioxane:MTBE (10 ml, 3.33 ml / g), and then dried in a vacuum oven at 60 °C to obtain the C104 bis(4-methylbenzenesulfonate) salt (5.05 g, 76.2% yield). Karl Fischer titration was performed to determine that the dried final product was anhydrous.

[0332] The relative amounts of each reagent used in this example are compared to the limiting reagent, which is C101.

[0333] (Example 5) Preparation of the monotosylate, bismesylate, monomesylate, monosulfate, and hemisulfate salts of C104 General procedure for the preparation of the monotosylate, bismesylate, monomesylate, monosulfate, and hemisulfate salts of C104 C104 (250 mg, 1.0 equivalent, limiting reagent) was dissolved in anisole (2 ml, 8 ml / g) with stirring. To the solution was added the defined stoichiometry (0.5 equivalent to 2.0 equivalents) of the desired acid. Then, ethyl acetate (2 ml, 8 ml / g) was added to the mixture as a poor solvent. The resulting slurry was filtered and dried in a vacuum oven at 50 °C.

[0334] Analytical data for the prepared salts was collected.

[0335] 5A. Monotosylate salt of C104

[0336]

Chemical formula

[0337]

Table 4

[0338] A sample of the monotosylate (p - toluenesulfonate) salt of C104 (as prepared by the procedure described in Example 5) was used to generate a powder X - ray diffraction pattern. The PXRD pattern for the monotosylate salt of C104 (as prepared by the procedure described in Example 5) is provided in Figure 3.

[0339] 5B. Bis(methanesulfonate) salt of C104

[0340]

Chemical formula

[0341]

Table 5

[0342] A sample of the bis(methanesulfonate) salt of C104 (as prepared by the procedure described in Example 5) was used to generate a powder X - ray diffraction pattern. The PXRD pattern for the bis(methanesulfonate) salt of C104 (as prepared by the procedure described in Example 5) is provided in Figure 4.

[0343] 5C. Monomethanesulfonate salt of C104

[0344]

Chemical formula

[0345]

Table 6

[0346] A sample of the monomesylate of C104 (as prepared by the procedure described in Example 5) was used to generate a powder X-ray diffraction pattern. The PXRD pattern for the monomesylate of C104 (as prepared by the procedure described in Example 5) is provided in Figure 5.

[0347] 5D. Monosulfate of C104

[0348]

Chemical formula

[0349]

Table 7

[0350] A sample of the monosulfate of C104 (as prepared by the procedure described in Example 5) was used to generate a powder X-ray diffraction pattern. The PXRD pattern for the monosulfate of C104 (as prepared by the procedure described in Example 5) is provided in Figure 6.

[0351] 5E. Hemisulfate of C104

[0352]

Chemical formula

[0353]

Table 8

[0354] A sample of the hemisulfate of C104 (as prepared by the procedure described in Example 5) was used to generate a powder X-ray diffraction pattern. The PXRD pattern for the hemisulfate of C104 (as prepared by the procedure described in Example 5) is provided in Figure 7.

[0355] (Example 6) Preparation of Methyl (S)-2-(Chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (C109)

[0356] [Chemical formula] Step 1. Preparation of Methyl (S)-4-amino-3-(oxetane-2-carboxamide)benzoate (C107) To a nitrogen-purged jacketed vessel equipped with overhead stirring, isopropanol (4.0 mL / g, 15.7 g, 20.0 mL) was added, followed by potassium hydroxide pellets (85 mass%, 1.65 equivalents, 3.28 g, 49.6 mmol). The mixture was heated to 40 °C over 30 minutes to dissolve most / all of the KOH. The mixture was then cooled to 10 °C at 0.5 °C / min. Then, C106 (88.4 mass%, 1.05 equivalents, 4.65 g, 31.6 mmol, ethyl (S)-oxetane-2-carboxylate) was added over 15 - 30 minutes. Once the addition was complete, the reaction mixture was stirred at 10 °C for 30 minutes, then warmed to 25 °C at 0.5 °C / min and the reaction mixture was left stirred at 25 °C. After 1 hour, a sample of the reaction mixture was taken for completion of the reaction (ester hydrolysis, this was carried out in CD3OD) 1(confirmed by \(^1H\) NMR). Triethylammonium hydrochloride (1.95 eq, 8.08 g, 58.7 mmol) was added to the reaction mixture at 25 °C, and the resulting mixture was stirred at 25 °C. After about 12 h at 25 °C, the reaction mixture was diluted with ethyl acetate (for reaction) (10.0 mL / g, 45.0 g, 50.0 mL). Then, C105 (5.0 g, 30.1 mmol, methyl 3,4-diaminobenzoate, limited reagent) was added to the reaction mixture. Then, diisopropylethylamine (3.00 eq, 11.7 g, 15.7 mL, 90.3 mmol) was added to the reaction mixture. Then, the reaction mixture was stirred at 25 °C for 30 min to homogenize / dissolve C105 and then cooled to 10 °C at 0.5 °C / min. Then, 50 wt% T3P (propanephosphonic anhydride, also known as PPAA, cyclic trimer) in EtOAc (ethyl acetate) (50 mass%, 1.50 eq, 28.7 g, 26.9 mL, 45.1 mmol) was added to the reaction mixture over 30 min while maintaining the temperature at 8 - 15 °C. The resulting reaction mixture was stirred at 10 °C. After 1 h, the reaction mixture was sampled for reaction completion (HPLC analysis for in-process monitoring). The reaction mixture was quenched with potassium bicarbonate (aqueous solution) (0.75 M) (10.0 mL / g, 50.0 mL). The reaction mixture was warmed to 25 °C, stirred to completely mix the phases, then the stirring was stopped and the phases were separated (two clear phases, no solids). The lower aqueous phase was drained from the vessel. Then, the aqueous phase was back-extracted with dichloromethane (10.0 mL / g, 66.3 g, 50.0 mL). After stirring to completely mix the phases, the stirring was stopped and the phases were separated (two clear phases, no solids). The phases were separated and the upper aqueous phase was discarded. The EtOAc and DCM extracts were combined. Then, the combined organic phase was washed with water (3.0 mL / g, 15.0 g, 15.0 mL). The phases were separated and the upper aqueous phase was discarded. The product solution was concentrated at a jacket temperature of 40 - 50 °C and 150 - 250 mbar (to about 12 mL / g, 60 mL). The vacuum was released and the mixture was self-seeded. Stirring at 40 - 45 °C for 15 min caused the material to start crystallizing. The product solution was concentrated at a jacket temperature of 40 - 50 °C and 150 - 250 mbar (to about 8 mL / g, 40 mL).The vacuum was released and the resulting fine suspension was held at 40 - 45 °C for 30 minutes and then cooled to 5 °C at 0.2 °C / min. The suspension was stirred at 5 °C for 1 hour. The suspension was filtered using vacuum filtration. The filter cake was washed with pre-cooled ethyl acetate (EtOAc, 2.0 mL / g, 9.00 g, 10.0 mL). After suction drying for 1 hour, the solid was dried under vacuum at 40 - 50 °C for 12 to 48 hours. When taken out of the oven, C107 was obtained as an off-white solid (crude weight: 4.62 g, crude yield: 61%).

[0357]

Table 9

[0358] The relative amounts of each reagent used in Step 1 of this example are compared to the limiting reagent which is C105.

[0359] Step 2. Preparation of Methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate (C108) Dichloromethane (DCM, 15.0 mL / g, 1590 g, 1200 mL) was added to a nitrogen-purged, jacketed vessel equipped with overhead stirring. C107 (80.00 g, 319.7 mmol; limiting reagent; methyl (S)-4-amino-3-(oxetane-2-carboxamide) benzoate) was added to the reactor vessel and stirred at 20 °C. Triethyl borate (3.00 equivalents, 140 g, 164 mL, 959 mmol) was then added to the reaction and stirred at 20 °C. Lithium borohydride (2 M) in tetrahydrofuran (1.25 equivalents, 180 g, 200 mL, 400 mmol) was added to the suspension over 30 minutes and stirred at 20 °C. Once the addition of LiBH4 was complete, the reaction was stirred at 20 °C and samples were taken periodically to confirm completion of the reaction. At the completion of the reaction, the reaction mixture was quenched slowly with water (5.0 mL / g, 400.0 g, 400.0 mL) over 30 minutes or more. The resulting mixture was stirred for over 2 hours and then allowed to phase separate. Stirring was stopped, the phases were separated, and the lower DCM phase was drained and transferred to a holding vessel. Next, phosphoric acid (0.5 M) in water (5.0 mL / g, 400.0 mL) was added to the DCM product solution and stirred at 20 °C for 30 minutes. Stirring was stopped, the phases were separated, and the lower DCM phase was drained and transferred to a holding vessel. Next, aqueous citric acid solution (0.5 M) in water (5.0 mL / g, 400.0 mL) was added to the DCM product solution and stirred at 20 °C for 30 minutes. Stirring was stopped, the phases were separated, and the lower DCM phase was drained and transferred to a holding vessel. Next, the DCM solution of C108 was held for analysis and proceeded to the next step without isolation of C108, assuming 100% yield for the next step (Step 3).

[0360] The relative amounts of each reagent used in Step 2 of this example are compared to the limiting reagent, which is C107.

[0361] Step 3. Preparation of methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (C109) Into a nitrogen-purged, jacketed vessel equipped with overhead stirring, the DCM solution of C108 from the previous step (Step 2) (total volume = ca. 1350 mL, theoretical = 75.5 g, 319.5 mmol, limiting reagent) was added and stirred at 20 °C. Under vacuum, the reaction contents were concentrated to a total volume of 3 to 4 mL / g (226 - 301 mL). The vacuum was released and then isopropanol (8.0 mL / g, 474.7 g) was charged. The mixture was concentrated again to a total volume of 3 to 4 mL / g (226 - 301 mL). To the mixture at 25 °C, 2-chloro-1,1,1-trimethoxyethane (1.05 eq, 51.87 g, 45.22 mL, 335.5 mmol) was added all at once. To the reaction, citric acid (0.01 eq, 0.6139 g, 3.195 mmol) was added all at once. The reaction was then heated to 50 °C. At the end of the reaction, after reducing the temperature to 30 °C, water (12.0 mL / g, 906.0 g, 906.0 mL) was added over 1 hour. At the end of the water addition, the mixture was a fine suspension, which was held at 30 °C for 30 minutes and then cooled to 5 °C. After 16 hours at 5 °C, the suspension was filtered and the solid was washed with IPA / water (1:9 v / v) (3.0 mL / g, 226.5 mL). The filter cake was suction dried under vacuum for 2 hours and then placed in a vacuum oven and dried at 40 °C for at least 12 hours to give 61.6 g (65% - 2 steps) of C109 as a white solid. 1 H NMR (600 MHz, CDCl3) δ 8.12 (s, 1H), 8.00 (d, 1H), 7.79 (d, 1H), 5.16 - 5.26 (m, 1H), 5.03 (s, 2H), 4.57 - 4.66 (m, 2H), 4.48 - 4.56 (m, 1H), 4.33 (m, 1H), 3.95 (s, 3H), 2.71 - 2.81 (m, 1H), 2.36 - 2.47 (m, 1H).

[0362] The relative amounts of each reagent used in Step 3 of this example are compared to the limiting reagent, which is C108.

[0363] (Example 7) (S)-2-((4-(6-((4-Cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (C110) Preparation

[0364] [Chemical formula] Acetonitrile (400 mL, 8 L / Kg) was added to the vessel, and the solvent was maintained at 25 °C. C104 bis(4-methylbenzenesulfonate) salt (57.3 g, 0.087 mol, 0.515 eq) was added to the vessel while maintaining the temperature at 25 °C ± 5.0 °C. Diisopropylethylamine (74.0 mL, 0.424, 2.5 eq) was slowly added to the vessel over at least 30 minutes while maintaining the temperature at 25 °C ± 5.0 °C. The second charge of C104 bis(4-methylbenzenesulfonate) salt (57.3 g, 0.087 mol, 0.515 eq) was added to the vessel while maintaining the temperature at 25 °C ± 5.0 °C. The second charge of diisopropylethylamine (74.0 mL, 0.424, 2.5 eq) was slowly added to the vessel over at least 30 minutes while maintaining the temperature at 25 °C ± 5.0 °C. C109 (50.0 g, 0.170 mol, 1.00 eq, limiting reagent) was added to the vessel. The reaction mixture was heated to 50 °C ± 5.0 °C over 30 minutes. This mixture was held at 50 °C ± 5.0 °C for at least 6 hours. A sample was taken for UPLC analysis. C110 seed crystals (0.5 g, 0.88 mmol, 0.01 g / g) were added to the vessel. The reaction mixture was granulated at 50 °C for 2 hours. Water (600 mL, 12 L / Kg) was added to the vessel over at least 1 hour while maintaining the temperature at 50 °C (acceptable temperature from 40 to 55 °C). It was cooled to 15 °C ± 5.0 °C over 3 hours (at a rate of 0.2 °C / min). The slurry was stirred at 15 °C for at least 8 hours. The reaction mixture was filtered. Water (175 mL, 3.5 mL / g) was added to an empty vessel. Acetonitrile (25 mL, 0.5 mL / g) was added to the vessel. The washing solution was stirred at 15 °C ± 5.0 °C. The washing solution of acetonitrile and water was transferred to the filter. The product was suction dried and loaded into a vacuum oven. The product was dried under vacuum at 50 °C for at least 12 hours (93.7 g, 97% yield). 1 H NMR (600 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.87 (d, 1H), 7.80 (d, 1H), 7.55 - 7.73 (m, 4H), 6.87 (d, 1H), 6.70 (d, 1H), 5.45 (s, 2H), 5.04 - 5.19 (m, 1H), 4.81 (dd, 1H), 4.66 (dd, 1H), 4.41 - 4.54 (m, 1H), 4.36 (dt, 1H), 3.94 (d, 1H), 3.86 (s, 3H), 3.76 (d, 1H), 2.97 (d, 1H), 2.82 (d, 1H), 2.63 - 2.77 (m, 1H), 2.49 - 2.63 (m, 1H), 2.37 - 2.46 (m, 1H), 2.18 - 2.29 (m, 1H), 2.05 - 2.18 (m, 1H), 1.47 - 1.82 (m, 4H). LC - MS(ES+): 570.5 (M + H).

[0365] The relative amounts of each reagent used in this example are compared to the limiting reagent, which is C109.

[0366] Using a sample of C110 (as prepared by the procedure described in Example 7), a powder X - ray diffraction pattern was generated and found to be crystalline. The observed PXRD pattern for crystalline C110 (designated as Form X herein) is provided in Figure 14, and the corresponding peak list is provided in Table E7 (along with the peaks whose relative intensity is 5% or more).

[0367] [Table 10 - 1]

[0368] [Table 10 - 2]

[0369] Preparation of C110 seed crystal material The seed crystal material of C110 can be prepared by the same procedures as described in this example, except for not using a seed crystal material (i.e., relying on self-nucleation).

[0370] (Example 7A) Alternative Preparation of Methyl (S)-2-((4-(6-((4-Cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (C110)

[0371] [Chemical formula] Methanol (1200 mL, 12 L / kg) was added to the vessel, and the solvent was maintained at 25 °C. C104 bis(4-methylbenzenesulfonate) salt (244.8 g, 373 mol, 1.1 eq) was added to the vessel at 25 °C. Diisopropylethylamine (297 mL, 1697 mol, 5.0 eq) was slowly added to the vessel over at least 15 minutes while maintaining the temperature at 25 °C. C109 (100.0 g, 339 mol, 1.0 eq, limiting reagent) was added to the vessel, and the reaction mixture was heated to 50 °C. The mixture was held at 50 °C for 15 minutes. C110 seed crystals (1.0 g, 1.8 mmol, 0.01 kg / kg) were added to the vessel. The mixture was held at 50 °C for at least 24 hours. Samples were taken for UPLC analysis. Water (400 mL, 4 L / kg) was added to the vessel over at least 1 hour while maintaining a temperature of 50 °C (an acceptable temperature range of 45 to 55 °C). The reaction mixture was granulated at 50 °C for 1 hour. It was cooled to 20 °C at a rate of 0.2 K / min. The slurry was stirred at 20 °C for at least 2 hours. The reaction product was filtered. Water (350 mL, 3.5 L / kg) was added to an empty vessel. Methanol (50 mL, 0.5 L / kg) was added to the vessel. The washing solution was stirred at 20 °C. The methanol and water washing solution was transferred to the filter. Water (350 mL, 3.5 L / kg) was added to an empty vessel. Methanol (50 mL, 0.5 L / kg) was added to the vessel. The washing solution was stirred at 20 °C. The methanol and water washing solution was transferred to the filter. The product was suction dried on the filter for 2 hours. The product was transferred to a vacuum oven and dried under vacuum at 50 °C for at least 16 hours (200.6 g, 100% yield). Compared to the process in Example 7, Process 7A reduced impurity IMP-A in the final product C110.

[0372]

Chemical formula

[0373] The relative amounts of each reagent used in this example are compared to the limiting reagent, which is C109.

[0374] Preparation of the C110 seed crystal material The C110 seed crystal material can be prepared by the same procedure as described in this example, except that no seed crystal material is used (i.e., relying on self-nucleation). Alternatively, the C110 seed crystal material can be prepared by the same procedure as described in Example 6.

[0375] (Example 8) Preparation of hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid

[0376]

Chemical formula

[0377] The relative amounts of each reagent used in this example are compared to the limiting reagent, which is C110.

[0378]

Table 11

[0379] A sample of the hemibarium salt of C111 (as prepared by the procedure described in Example 8) was used to generate a powder X-ray diffraction pattern. The PXRD pattern for the hemibarium salt of C111 (as prepared by the procedure described in Example 8) is provided in Figure 8.

[0380] (Example 9) (S)-2-((4-(6-((4-Cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, tris salt (tris salt of C111) Preparation

[0381] [Chemical formula] 1.5 mL / g (105 mL) of H2O was added to the 2 L reactor when it was ready. Subsequently, 7 mL / g (490 mL) of MeCN was added to the vessel all at once. 10.53 g (61.44 mmol, 0.5 equivalent) of Ba(OH)2 was added to the vessel as a solid, and the vessel was stirred to ensure a suspension of barium hydroxide. To the same vessel, 70.0 g (122.9 mmol, limiting reagent) of C110 was added as a solid all at once, along with a small amount of MeCN used to rinse the addition funnel. Subsequently, the reactants were heated to an internal temperature of 55 °C at a rate of 1 °C / min. The reactants were held under N2(g) for a total of 24 hours under these conditions. Samples of the reactants were taken to confirm the completion of the reaction (LC). Then, upon completion of the reaction, the reaction mixture was cooled to an internal temperature of 25 °C at a rate of 1 °C / min. 3.5 mL / g (245 ml) of H2O was added to this slurry. 1 ml / g (70 ml) of toluene was added to the slurry. 1.5 equivalents (10.56 mL, 184.3 mmol) of acetic acid was added to this slurry. The resulting mixture was stirred at 25 °C for 30 minutes. The phases were separated. The organic solution was heated to 45 °C. To the organic solution, 1.20 equivalents (18.05 g, 147.5 mmol) of 2-amino-2-(hydroxymethyl)-1,3-propanediol was added as a solution in water (1 mL / g, 70 mL). A seed crystal charge of 0.004 g / g of C111 tris salt was added to the solution (0.280 g, 0.413 mmol, 0.003 molar equivalent). The reaction slurry was held at 45 °C for 60 minutes. The reactants were cooled to 25 °C at a rate of 1 °C / min and then held for 60 minutes. The reactants were heated to 45 °C at a rate of 1 °C / min and then held for 60 minutes. The reactants were cooled to 20 °C at a rate of 1 °C / min and held overnight. The slurry was isolated by filtration, and the liquid was drawn off to the top of the cake. The cake was washed with a first MEK (3 mL / g, 210 ml, methyl ethyl ketone) wash solution and the liquid was drawn off to the top of the cake. The cake was washed with a second MEK (3 mL / g, 210 ml) wash solution and the liquid was drawn off to the top of the cake. The resulting cake was dried by suction for 30 minutes. The solid (C111 tris salt) was unloaded and dried overnight in a vacuum oven at 50 °C (73.01 g, 107.9 mmol, approximately 87% yield).

[0382] The final product (C111 tris salt) prepared in this example contains a reduced amount of impurity IMP-2 compared to the method in Example 4A-01 of U.S. Patent No. 10,208,019.

[0383]

Chemical formula

[0384] The relative amounts of each reagent used in this example are compared to the limiting reagent, which is C110.

[0385] Preparation of the tris salt of the C111 seed crystal material The seed crystal material of the tris salt of C111 can be prepared by the same procedure as described in this example, except that no seed crystal material is used (i.e., relying on self-nucleation).

[0386] (Example 10) (S)-2-((4-(6-((4-Cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid tris salt (tris salt of C111) alternative preparation

[0387]

Chemical formula

[0388] The final product (C111 tris salt) prepared in this example contains a reduced amount of impurity IMP-2 compared to the method in Example 4A-01 of U.S. Patent No. 10,208,019.

[0389] The relative amounts of each reagent used in this example are compared to the limiting reagent, which is C110.

[0390] Preparation of the tris salt of C111 seed material The seed material of the tris salt of C111 can be prepared by the same procedure as described in this example, except that no seed material is used (i.e., relying on self-nucleation). Alternatively, the seed material of the tris salt of C111 can be prepared by the same procedure as described in Example 9.

[0391] (Example 11) Preparation of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, Form 1 of the tris salt (Form 1 of the tris salt of C111)

[0392] [Chemical formula] DMSO (16 mL, 8 L / Kg) was added to the vessel and the solvent was maintained at 25 °C. Water (2 mL, 1 L / Kg) was added to the vessel and the solvent was maintained at 25 °C. C111 tris salt (2.0 g, 2.96 mmol, 1.0 equivalent, limiting reagent) was added to the vessel while maintaining a temperature of 25 °C ± 5.0 °C. The reaction mixture was heated to 65 °C over 30 minutes. The reaction mixture was maintained at 65 °C for at least 30 minutes. Water (2 mL, 1 L / Kg) was slowly added to the vessel over 30 minutes while maintaining a temperature of 65 °C. The reaction mixture was maintained at 65 °C for at least 30 minutes. Seed crystal C111 tris salt (10 mg, 0.01 mmol, 0.005 g / g) was added to the vessel while maintaining a temperature of 65 °C. The reaction mixture was maintained at 65 °C for at least 1 hour. The reaction mixture was cooled to 30 °C at a rate of 0.2 °C / min. The reaction mixture was maintained at 30 °C for 1 hour. The reaction mixture was heated to 45 °C over 30 minutes. The reaction mixture was maintained at 45 °C. The reaction mixture was cooled to 15 °C at a rate of 0.2 °C / min. The reaction mixture was granulated at 15 °C for at least 8 hours. The batch was filtered and dried by suction. Butan-2-one (or methyl ethyl ketone, MEK) (6 mL, 3 L / Kg) was added to the vessel and the solvent was cooled to 15 °C. The cake wash was transferred to the filter and the batch was dried by suction. The product was transferred to an oven and dried at 55 °C for at least 8 hours.

[0393] The final product (C111 tris salt) prepared in this example contains reduced amounts of impurities IMP-1 and IMP-2 compared to the method in Example 4A-01 of U.S. Patent No. 10,208,019.

[0394]

Chemical formula

[0395] Preparation of Form 1 of the Tris Salt of C111 Seed Crystal Material The seed crystal material of Form 1 of the tris salt of C111 can be prepared by a procedure similar to that described in this example, except that no seed crystal material is used (i.e., relying on self-nucleation).

[0396] (Example 12) (S)-2-((4-(6-((4-Cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, alternative preparation of tris salt form 1 (tris salt form 1 of C111)

[0397] [Chemical formula] 4.5 g of C111 tris salt (limited reagent), 18.75 ml of THF (4.17 L / kgW) and 6.25 ml of water (1.39 L / kgW) were charged into a vessel and stirred. The slurry was heated to 55 °C to confirm dissolution. Then, the mixture was cooled to 49 °C at 1 K / min, and when the contents were checked, a solution remained. 1% of the seed crystal of C111 tris salt was added to the solution. After seeding, the mixture was held at 49 °C for 1 hour and the slurry was checked. The mixture was cooled to 35 °C at 0.1 - 0.2 K / min and then held at 35 °C for 1 hour. 25 ml of acetonitrile (5.56 L / kgW) was added over 1.5 hours. The resulting mixture was held at 35 °C for 1 hour and then cooled to 10 °C at 0.2 K / min. The mixture was held at 10 °C for at least 8 hours. A sample of the slurry was taken to check the particle size. If the D90 of the particles in the slurry was 125 microns or more, high-shear wet grinding was carried out. The sample collection and wet grinding steps for particle size confirmation were repeated several times until the D90 of the particles in the slurry was less than 125 microns. Then, the slurry was filtered under pressure and washed with 2 × 10 ml of MEK (2 × 2.22 L / kg) while stirring. The material was transferred to a dryer and dried under reduced pressure at a jacket temperature of 55 °C (4.14 g, 92%).

[0398] The final product (C111 tris salt) prepared by the method in this example contains reduced amounts of impurities IMP-1, IMP-2, and IMP-3 compared to the method in Example 4A-01 of U.S. Patent No. 10,208,019. Further, the process in this example provides the final product with a larger particle size, and the particle size was relatively uniform (e.g., compared to the method in Example 4A-01 of U.S. Patent No. 10,208,019).

[0399] [Chemical formula]

[0400] Preparation of Form 1 of the Tris Salt of C111 Seed Crystal Material The seed crystal material of Form 1 of the tris salt of C111 can be prepared by the same procedure as described in this example, except for not using a seed crystal material (i.e., relying on self-nucleation). Alternatively, the seed crystal material of Form 1 of the tris salt of C111 can be prepared by the same procedure as described in Example 11.

[0401] A sample of Form 1 of the tris salt of C111 (as prepared by the procedure as described in Example 11 or 12) was used to generate a powder X-ray diffraction pattern. The PXRD pattern for Form 1 of the tris salt of C111 (as prepared by the procedure as described in Example 11 or 12) is provided in Figure 9, and the corresponding peak list is provided in Table E12 (along with the peaks whose relative intensity is 5% or more).

[0402] [Table 12]

[0403] (Example 13) Solid-State NMR Analysis of Form 1 of the Tris Salt of C111 Solid-state NMR (ssNMR) analysis [using a sample of Form 1 of the tris salt of C111 as prepared by the procedure as described in Example 11 or 12] was performed on a Bruker Avance III HD 400 MHz ( 1 H frequency) NMR spectrometer. A 4 mm magic angle spinning (MAS) probe was used at a MAS speed of 10 kHz for 13 C analysis. 19 F spectra were recorded using a 3.2 mm MAS probe at a spin speed of 20 kHz. 15 N ssNMR analysis was performed on a Bruker Avance Neo 400 MHz NMR spectrometer equipped with a 4 mM MAS probe at a spin speed of 20 kHz. All spectra were acquired at a temperature adjusted to 20 °C.

[0404] Using TOSS spin sideband suppression 13 C cross-polarization (CP) measurements were recorded with a CP contact time of 4 milliseconds and a recycle delay of 40 seconds (see Figure 10 and Table E13-a). A phase-modulated proton decoupling field of approximately 100 kHz was applied during spectrum acquisition. Carbon spectrum referencing was done relative to undiluted tetramethylsilane by setting the high-frequency signal from an external sample of adamantane to 38.5 ppm.

[0405] 19 F spectra were collected by direct excitation using proton decoupling and a recycle delay of 120 seconds (see Figure 11 and Table E13-b). Spectrum referencing was done for CFCl3 by setting the resonance from an external sample of 50% v / v trifluoroacetic acid in H2O to -76.54 ppm.

[0406] 15 N CP spectra were recorded with a CP contact time of 10 milliseconds and a recycle delay of 3 seconds (see Figure 12 and Table E13-c). Nitrogen spectrum referencing was done relative to undiluted nitromethane by setting the signal from an external sample of glycine to -346.8 ppm.

[0407]

Table 13-1

[0408]

Table 13-2

[0409]

Table 14

[0410]

Table 15

[0411] The peak positions and relative intensities were obtained using the ACD Labs Spectrus Processor 2019 software. 13 C, 15 N and 19 The error in the peak positions reported in the 19F ssNMR data is estimated to be ±0.2 ppm. The ssNMR intensities can vary depending on the experimental parameter settings and the thermal history of the sample.

[0412] (Example 14) Raman spectroscopic analysis of Form 1 of the tris salt of C111 The Raman spectrum [using a sample of Form 1 of the tris salt of C111 prepared by the procedure as described in Example 11 or 12] was collected using the RAM II FT-Raman module (Bruker Optik GmbH) attached to a Vertex 70 spectrometer. The instrument was equipped with a 1064 nm solid-state (Nd:YAG) laser and a liquid nitrogen-cooled germanium detector. Prior to data acquisition, the instrument performance and calibration verification were performed using a white light source and polystyrene and naphthalene reference substances.

[0413] Samples were prepared and analyzed in cut NMR tubes. A sample spinner (Ventacon, UK) was used during measurement to maximize the volume of material exposed to the laser during data collection. The Raman signal backscattered from the sample was optimized, and data were collected at a spectral resolution of 2 cm -1 with a laser power output of 500 mW. A Blackman-Harris 4-term apodization function was applied to minimize spectral aberration. Spectra were generated with an appropriate number of scans adjusted as needed to ensure a reasonable signal-to-noise between 3500 and 50 cm -1 .

[0414] The spectra were normalized by setting the intensity of the most intense peak to 2.00. Peaks were then identified using the automatic peak picking function in OPUS v8.2 software (Bruker Optik GmbH) with the sensitivity set to 2%. Peak positions and relative peak intensities were extracted and tabulated. The variability in peak positions using this experimental setup is within ±2 cm -1 .

[0415] Figure 13 shows a representative FT-Raman spectrum of Form 1 of the tris salt of C111, and Table E14 shows an FT-Raman peak list for Form 1 of the tris salt of C111.

[0416]

Table 16-1

[0417]

Table 16-2

[0418] Form 1 of the tris salt of C111 is, for example, the powder X-ray diffraction (PXRD) data, solid-state nuclear magnetic resonance (ssNMR) data (e.g., 13 C ssNMR data, 15 N ssNMR data, and / or 19It can be identified by its unique solid state footprint with respect to 19F ssNMR), and / or FT-Raman spectroscopy data.

[0419] Form 1 of the tris salt of C111 has a powder X-ray diffraction pattern (PXRD) that includes one or two peaks selected from those at 14.3 ± 0.2°, 17.5 ± 0.2°, and 18.0 ± 0.2° with respect to 2θ (CuKα radiation source, wavelength 1.5406 Å). In some embodiments, form 1 of the tris salt of C111 has a PXRD that includes one peak at 14.3 ± 0.2° with respect to 2θ.

[0420] In some embodiments, form 1 of the tris salt of C111 has a PXRD that includes peaks at 14.3 ± 0.2°, 17.5 ± 0.2°, 18.0 ± 0.2°, and 23.4 ± 0.2° with respect to 2θ.

[0421] In some embodiments, form 1 of the tris salt of C111 has PXRD peaks at 14.3 ± 0.2°, 17.5 ± 0.2°, 18.0 ± 0.2°, 23.4 ± 0.2°, and 24.7 ± 0.2° with respect to 2θ. In further embodiments, form 1 of the tris salt of C111 has a powder X-ray diffraction pattern (PXRD) that is substantially the same as FIG. 9 (FIG. 9).

[0422] In some embodiments, form 1 of the tris salt of C111 includes one peak selected from those at 171.0 ± 0.2 ppm and 141.3 ± 0.2 ppm with respect to chemical shift 13 and has a 13C ssNMR spectrum. In some further embodiments, form 1 of the tris salt of C111 13 has a 13C ssNMR spectrum.

[0423] In some embodiments, form 1 of the tris salt of C111 includes peaks at 171.0 ± 0.2 ppm and 141.3 ± 0.2 ppm with respect to chemical shift 13 and has a 13C ssNMR spectrum.

[0424] In some embodiments, Form 1 of the tris salt of C111 includes peaks at 171.0 ± 0.2 ppm, 141.3 ± 0.2 ppm, and 64.0 ± 0.2 ppm with respect to chemical shift. 13 It has a C ssNMR spectrum.

[0425] In some embodiments, Form 1 of the tris salt of C111 includes peaks at 171.0 ± 0.2 ppm, 141.9 ± 0.2 ppm, 141.3 ± 0.2 ppm, 120.7 ± 0.2 ppm, and 64.0 ± 0.2 ppm with respect to chemical shift. 13 It has a C ssNMR spectrum. In further embodiments, Form 1 of the tris salt of C111 has a C ssNMR spectrum that is substantially the same as FIG. 10 (FIG. 10). 13 It has a C ssNMR spectrum.

[0426] In some embodiments, Form 1 of the tris salt of C111 includes one peak at -118.8 ± 0.2 ppm with respect to chemical shift. 19 It has an F ssNMR spectrum. In further embodiments, the crystalline form has an F ssNMR spectrum that is substantially the same as FIG. 11 (FIG. 11). 19 It has an F ssNMR spectrum.

[0427] In some embodiments, Form 1 of the tris salt of C111 includes one peak at -339.9 ± 0.2 ppm or -223.4 ± 0.2 ppm with respect to chemical shift. 15 It has an N ssNMR spectrum.

[0428] In some embodiments, Form 1 of the tris salt of C111 includes one peak at -339.9 ± 0.2 ppm with respect to chemical shift. 15 It has an N ssNMR spectrum.

[0429] In some embodiments, Form 1 of the tris salt of C111 includes one peak at -223.4 ± 0.2 ppm with respect to chemical shift. 15 It has an N ssNMR spectrum.

[0430] In some embodiments, Form 1 of the tris salt of C111 includes peaks at -339.9 ± 0.2 ppm and -223.4 ± 0.2 ppm with respect to chemical shift. 15 has an N ssNMR spectrum. In further embodiments, Form 1 of the tris salt of C111 has an N ssNMR spectrum that is substantially the same as that of FIG. 12 (FIG. 12). 15 has an N ssNMR spectrum.

[0431] In some embodiments, Form 1 of the tris salt of C111 has an FT-Raman spectrum that includes one or two peaks selected from those at a wavenumber (cm -1 ) of 1371 ± 2 cm -1 , 430 ± 2 cm -1 , and 416 ± 2 cm -1 .

[0432] In some embodiments, Form 1 of the tris salt of C111 has an FT-Raman spectrum that includes peaks at a wavenumber (cm -1 ) of 1371 ± 2 cm -1 , 430 ± 2 cm -1 , and 416 ± 2 cm -1 .

[0433] In some embodiments, Form 1 of the tris salt of C111 has an FT-Raman spectrum that includes peaks at a wavenumber (cm -1 ) of 1371 ± 2 cm -1 , 430 ± 2 cm -1 , 416 ± 2 cm -1 , and 3026 ± 2 cm -1 . In further embodiments, in some embodiments, Form 1 of the tris salt of C111 has an FT-Raman spectrum that is substantially the same as that of FIG. 13 (FIG. 13).

[0434] (Example AA) CHO GLP-1R Clone H6-Assay 1 GLP-1R-mediated agonist activity was determined in a cell-based functional assay using a HTRF (Homogeneous Time-Resolved Fluorescence) cAMP detection kit (cAMP HI Range Assay Kit; CisBio catalog number 62AM6PEJ) that measures cAMP levels in cells. The method is a competitive immunoassay between native cAMP produced by the cells and exogenous cAMP labeled with the dye d2. The tracer binding is visualized by a mAb anti-cAMP labeled with cryptate. The specific signal (i.e., energy transfer) is inversely proportional to the concentration of cAMP in either the standard or experimental sample.

[0435] The human GLP-1R coding sequence (NCBI reference sequence NP_002053.3 including the naturally occurring variant Gly168Ser) was subcloned into pcDNA3 (Invitrogen), and a cell line stably expressing the receptor was isolated (designated clone H6). 125 I-GLP-1 7-36 (Perkin Elmer) using a saturation binding assay (filtration assay procedure) showed that the plasma membrane derived from this cell line expressed a high GLP-1R density (K d : 0.4 nM, B max : 1900 fmol / mg protein).

[0436] Cells were removed from cryopreservation, resuspended in 40 mL of Dulbecco's Phosphate Buffered Solution (DPBS - Lonza catalog number 17 - 512Q), and centrifuged at 800×g for 5 minutes at 22°C. The cell pellet was then resuspended in 10 mL of growth medium [DMEM / F12 1:1 mixture supplemented with HEPES, L - Gln, 500 mL (DMEM / F12 Lonza catalog number 12 - 719F), 10% heat - inactivated fetal bovine serum (Gibco catalog number 16140 - 071), 5 mL of 100× penicillin - streptomycin (Gibco catalog number 15140 - 122), 5 mL of 100× L - glutamine (Gibco catalog number 25030 - 081), and 500 μg / mL geneticin (G418) (Invitrogen 10131035)]. A 1 mL sample of the cell suspension in growth medium was counted using a Becton Dickinson hemocytometer to determine cell viability and cell count per mL. The remaining cell suspension was then adjusted with growth medium and 2000 viable cells per well were delivered using a Matrix CombiMultiDrop reagent dispenser and dispensed into a white 384 - well tissue culture - treated assay plate (Corning 3570). The assay plate was then incubated in a humidified environment at 37°C in 5% carbon dioxide for 48 hours.

[0437] Each compound to be tested at various concentrations (in DMSO) was diluted in assay buffer (HBSS supplemented with calcium / magnesium (Lonza / BioWhittaker catalog number 10 - 527F) / 0.1% BSA (Sigma Aldrich catalog number A7409 - 1L) / 20 mM HEPES (Lonza / BioWhittaker catalog number 17 - 737E)) containing 100 μM 3 - isobutyl - 1 - methylxanthine (IBMX; Sigma catalog number I5879). The final DMSO concentration was 1%.

[0438] After 48 hours, the growth medium was removed from the assay plate wells and the cells were treated with 20 μL of serial diluted compound in assay buffer for 30 minutes at 37 °C in a humidified environment with 5% carbon dioxide. After 30 minutes of incubation, 10 μL of labeled d2 cAMP and 10 μL of anti-cAMP antibody (both diluted 1:20 in cell lysis buffer; as described in the manufacturer's assay protocol) were added to each well of the assay plate. The plate was then incubated at room temperature and after 60 minutes, changes in the HTRF signal were read using an Envision 2104 multilabel plate reader with excitation at 330 nm and emission at 615 and 665 nm. The raw data was converted to nM cAMP by interpolation from a cAMP standard curve (as described in the manufacturer's assay protocol), and the percent effect relative to the saturating concentration of the full agonist GLP-1 7-36 (1 μM) was determined. EC 50 Determinations were made from agonist dose-response curves analyzed by a curve fitting program using a four-parameter logistic dose-response equation.

[0439] (Example BB) CHO GLP-1R Clone C6 - Assay 2 GLP-1R-mediated agonist activity was determined in a cell-based functional assay using an HTRF (homogeneous time-resolved fluorescence) cAMP detection kit (cAMP HI Range Assay Kit; Cis Bio catalog number 62AM6PEJ) that measures cAMP levels in cells. The method is a competitive immunoassay between native cAMP produced by the cells and exogenous cAMP labeled with the dye d2. Tracer binding is visualized by an mAb anti-cAMP labeled with cryptate. The specific signal (i.e., energy transfer) is inversely proportional to the concentration of cAMP in either the standard or experimental samples.

[0440] The human GLP-1R coding sequence (NCBI reference sequence NP_002053.3 including the naturally occurring variant Leu260Phe) was subcloned into pcDNA5-FRT-TO, and a clonal CHO cell line stably expressing low receptor density was isolated using the Flp-In™ T-Rex™ system as described by the manufacturer (ThermoFisher). 125 Saturation binding assays (filtration assay procedure) using I-GLP-1 (Perkin Elmer) showed that the plasma membrane derived from this cell line (designated clone H6) expressed a lower GLP-1R density (K d : 0.3 nM, B max : 240 fmol / mg protein) compared to the clone H6 cell line.

[0441] Cells were removed from cryopreservation, resuspended in 40 mL of Dulbecco's Phosphate Buffered Solution (DPBS - Lonza catalog number 17 - 512Q), and centrifuged at 800×g for 5 minutes at 22°C. The DPBS was aspirated, and the cell pellet was resuspended in 10 mL of complete growth medium (DMEM:F12 1:1 mixture supplemented with HEPES, L - Gln, 500 mL (DMEM / F12 Lonza catalog number 12 - 719F), 10% heat - inactivated fetal bovine serum (Gibco catalog number 16140 - 071), 5 mL of 100× penicillin - streptomycin (Gibco catalog number 15140 - 122), 5 mL of 100× L - glutamine (Gibco catalog number 25030 - 081), 700 μg / mL hygromycin (Invitrogen catalog number 10687010) and 15 μg / mL blasticidin (Gibco catalog number R21001)). A 1 mL sample of the cell suspension in the growth medium was counted using a Becton Dickinson hemocytometer to determine cell viability and cell count per mL. The remaining cell suspension was then adjusted with growth medium and 1600 viable cells per well were delivered using a Matrix CombiMultiDrop reagent dispenser and dispensed into a white 384 - well tissue culture - treated assay plate (Corning 3570). The assay plate was then incubated at 37°C for 48 hours in a humidified environment (95% O2, 5% CO2).

[0442] Each compound to be tested at various concentrations (in DMSO) was diluted in assay buffer [HBSS supplemented with calcium / magnesium (Lonza / BioWhittaker catalog number 10 - 527F) / 0.1% BSA (Sigma Aldrich catalog number A7409 - 1L) / 20 mM HEPES (Lonza / BioWhittaker catalog number 17 - 737E)] containing 100 μM 3 - isobutyl - 1 - methylxanthine (IBMX; Sigma catalog number I5879). The final DMSO concentration in the compound / assay buffer mixture was 1%.

[0443] After 48 hours, the growth medium was removed from the assay plate wells and the cells were treated with 20 μL of serially diluted compound in assay buffer for 30 minutes at 37 °C in a humidified environment (95% O2, 5% CO2). After 30 minutes of incubation, 10 μL of labeled d2 cAMP and 10 μL of anti-cAMP antibody (both diluted 1:20 in cell lysis buffer; as described in the manufacturer's assay protocol) were added to each well of the assay plate. The plate was then incubated at room temperature and after 60 minutes, changes in the HTRF signal were read using an Envision 2104 multilabel plate reader with excitation at 330 nm and emission at 615 and 665 nm. The raw data was converted to nM cAMP by interpolation from a cAMP standard curve (as described in the manufacturer's assay protocol) and the percent effect relative to the saturating concentration of the full agonist GLP-1 (1 μM) included in each plate was determined. EC 50 Determinations were made from agonist dose-response curves analyzed by a curve fitting program using a four-parameter logistic dose-response equation.

[0444] In Table BB-1, the assay data are presented as geometric mean (EC 50 ) and arithmetic mean (Emax) to two (2) significant figures based on the number of replicates included.

[0445]

Table 17

[0446] All patents, patent applications, and references mentioned in this specification are hereby incorporated by reference in their entirety.

Claims

1. A process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, comprising: 【Chemical 1】 (a1) reacting tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate with 3-fluoro-4-(hydroxymethyl)benzonitrile in a solvent system in the presence of a palladium catalyst, a base, and a phosphorus ligand to form tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate, wherein the amount of 3-fluoro-4-(hydroxymethyl)benzonitrile is from about 1.0 to about 1.1 molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate; (a2) adding ethanol to the reaction mixture from step (a1); (a3) filtering the resulting reaction mixture from step (a2); (b1) adding p-toluenesulfonic acid monohydrate to the filtrate from step (a3), thereby reacting tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate with p-toluenesulfonic acid monohydrate to form the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, wherein the amount of p-toluenesulfonic acid monohydrate is from about 2.0 to about 3.0 molar equivalents relative to tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate; and (b2) isolating the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile from step (b1). A process as claimed in claim 1, which comprises the steps of:

2. A process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, comprising: 【Chemical Formula 2】 (a1) Reacting tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate with 3-fluoro-4-(hydroxymethyl)benzonitrile in a solvent system in the presence of a palladium catalyst, a base, and a phosphorus ligand to form tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate, where the amount of 3-fluoro-4-(hydroxymethyl)benzonitrile is from about 1.0 to about 1.1 molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate, and (a2) Adding water, ethyl acetate, and ethanol to the reaction mixture upon completion of the reaction in step (a1); (a3) Separating the organic phase from the aqueous phase from step (a2); (b1) Adding p-toluenesulfonic acid monohydrate to the separated organic phase from step (a3), thereby reacting tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate with p-toluenesulfonic acid monohydrate to form the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, where the amount of p-toluenesulfonic acid monohydrate is from about 2.0 to about 3.0 molar equivalents relative to tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate, and (b2) Isolating the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile from step (b1) A process comprising.

3. A process for preparing the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, comprising [Chemical Formula 3] (a1) React tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate with 3-fluoro-4-(hydroxymethyl)benzonitrile in a solvent system in the presence of a copper catalyst and a ligand to form tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate, wherein the amount of 3-fluoro-4-(hydroxymethyl)benzonitrile is about 1.1 to about 1.3 (e.g., 1.2) molar equivalents relative to tert-butyl 4-(6-chloropyridin-2-yl)piperidine-1-carboxylate; (a2) Filter the reaction mixture from step (a1), wherein filtering further comprises washing with methyl tert-butyl ether (MTBE); (a3) Concentrate the filtrate from step (2) to obtain tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate; (b1) Dissolve tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate from step (a3) in a solvent system comprising 1,4-dioxane and MTBE to form a solution, then add p-toluenesulfonic acid monohydrate to the solution, whereby tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate is reacted with p-toluenesulfonic acid monohydrate to form the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-((((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, wherein the amount of p-toluenesulfonic acid monohydrate is about 2.0 to about 2.5 molar equivalents relative to tert-butyl 4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidine-1-carboxylate; (b2) Isolate the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-((((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile from step (b1); (5) A process comprising the above steps. (6) Claim 4 Anhydrous 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt, Monohydrate of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile bis(4-methylbenzenesulfonate) salt, Monotosylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, Bismesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, Monomesylate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, Monosulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, Hemisulfate salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile, Crystalline methyl (S)-2-((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, or Hemibarium salt of (S)-2-((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid is a compound.

5. A process for preparing methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, comprising 【Chemical Formula 4】 (a1) preparing methyl (S)-4-amino-3-(oxetane-2-carboxamido)benzoate, (a2) reducing methyl (S)-4-amino-3-(oxetane-2-carboxamido)benzoate in the presence of a reducing reagent to form methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate, Step of reacting methyl (S)-4-amino-3-((oxetan-2-ylmethyl)amino)benzoate with 2-chloro-1,1,1-trimethoxyethane in the presence of an acid to form methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate A process comprising the same. **Claim 6** A process for preparing methyl (S)-2-(((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, [Chemical Formula 5] (a1) Reacting methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-((((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in a solvent system containing acetonitrile in the presence of diisopropylethylamine to form methyl (S)-2-(((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, wherein the amount of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-((((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is about 1.1 to about 1.5 molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and the amount of diisopropylethylamine is about 4.0 to about 6.0 molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate A process comprising the same. **Claim 7** A process for preparing methyl (S)-2-(((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, 【Chemical Formula 6】 Step of reacting (a1) methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile in a solvent system containing methanol in the presence of diisopropylethylamine to form methyl (S)-2-(((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, wherein the amount of the bis(4-methylbenzenesulfonate) salt of 3-fluoro-4-(((6-(piperidin-4-yl)pyridin-2-yl)oxy)methyl)benzonitrile is from about 1.0 to about 1.2 molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate, and the amount of diisopropylethylamine is from about 4.0 to about 6.0 molar equivalents relative to methyl (S)-2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate A process comprising the above step Claim 8 A process for preparing the hemibarium salt of (S)-2-(((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, comprising 【Chemical Formula 7】 (a1) Reacting methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with barium hydroxide in a solvent system comprising an organic solvent and water to form the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, wherein the amount of barium hydroxide is from about 0.5 to about 0.6 molar equivalents relative to methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate; (a2) Optionally, isolating the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid from step (a1); A process comprising the above steps. **Claim 9** A process for preparing the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, comprising: [Chemical Formula 8] Reacting methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with barium hydroxide in a solvent system containing acetonitrile and water to form the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, wherein the amount of barium hydroxide is from about 0.5 to about 0.6 molar equivalents relative to methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate; Adding water, a water-immiscible organic solvent, and an organic acid to the reaction mixture in step (a1), mixing the resulting mixture to form (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid; Separating the organic phase / solution from the aqueous phase / solution of the mixture from step (a2); Adding 2-amino-2-(hydroxymethyl)-1,3-propanediol to the separated organic phase from step (a3), thereby reacting 2-amino-2-(hydroxymethyl)-1,3-propanediol and (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid to form the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, wherein the reaction mixture is held at a holding temperature of from about 35 °C to about 55 °C; The seed crystalline material of the tris salt of (a5) (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid is added to the reaction mixture from step (a4) to form a slurry, where the step of holding the slurry at the holding temperature for a time longer than about 1 minute, and the step of cooling the slurry from step (a5) to a temperature of about 20 °C to about 30 °C and holding the slurry at that temperature for a time longer than about 1 minute, and optionally, the step of heating the slurry from step (a6) to the holding temperature and holding the slurry at the holding temperature for a time longer than about 1 minute, and optionally, the step of cooling the slurry from step (a7) to a temperature of about 15 °C to about 25 °C and holding the slurry at that temperature for a time longer than about 1 minute, and the step of isolating the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid in the slurry from step (a6) or (a8) and a process comprising. Claim 10 A process for preparing the tris salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, comprising 【Chemical Formula 9】 Step (a1): React methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate with barium hydroxide in a solvent system containing acetone and water to form the hemibarium salt of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, where the amount of barium hydroxide is from about 0.5 to about 0.6 molar equivalents relative to methyl (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate. Step (a2): Add water, a water-immiscible organic solvent, and an organic acid to the reaction mixture from step (a1), and mix the resulting mixture to form (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid. Step (a3): Optionally, separate the organic phase from the aqueous phase of the mixture from step (a2) and add methanol to the separated organic phase. (a4) Add 2-amino-2-(hydroxymethyl)-1,3-propanediol to the separated organic phase from step (a3), whereby 2-amino-2-(hydroxymethyl)-1,3-propanediol and (S)-2-(((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid are reacted to form the tris salt of (S)-2-(((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid, wherein the reaction mixture is held at a holding temperature of from about 35 °C to about 55 °C; (a5) Add a seed crystalline material of the tris salt of (S)-2-(((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid to the reaction mixture from step (a4) to form a slurry, wherein the slurry is held at the holding temperature for a time longer than about 1 minute; (a6) Cool the slurry from step (a5) to a temperature of from about 15 °C to about 20 °C and hold the slurry at that temperature for a time longer than about 1 minute; (a7) Isolate the tris salt of (S)-2-(((4-(6-(((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid in the slurry from step (a6). A process comprising the above steps. [

11. ] A method for preparing Form 1 of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, Suspending the tris salt of 2 - [(4 - {6 - [(4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [(2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid in a solvent system, where the solvent system consists of dimethyl sulfoxide (DMSO) and water, the volume ratio of DMSO:water is from about 10:1 to about 6:1, and the ratio of water (volume) to the tris salt of 2 - [(4 - {6 - [(4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [(2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid (weight) is from about 0.8 mL / g to about 1.2 mL / g at a temperature from about 20 °C to about 35 °C; heating the suspension from step (a) to a temperature increase from about 60 °C to about 70 °C to form a solution, and then mixing the solution for a time longer than about 1 minute during the temperature increase; slowly adding water to the solution from step (b) while maintaining the reaction mixture as a solution, where the amount of water added is approximately the same as the water used in step (a), and then holding the resulting solution for a time longer than about 1 minute during the temperature increase; seeding the solution with the crystalline form 1 material of the tris salt of 2 - [(4 - {6 - [(4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [(2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid while maintaining the temperature at the temperature increase, where the amount of the seed crystal is at least about 0.5 wt% of the tris salt of 2 - [(4 - {6 - [(4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [(2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid used in step (a), and then holding the resulting mixture for a time longer than about 1 minute during the temperature increase; optionally, slowly cooling the temperature of the mixture from step (d) to about 30 °C, and then holding the mixture at that temperature for a time longer than about 1 minute; (f)Optionally, slowly heat the mixture from step (e) from about 40 °C to about 50 °C, and then hold the mixture at that temperature for a time longer than about 1 minute; (g)Slowly cool the temperature of the mixture from step (d) or step (f) [when steps (e) and (f) are carried out] to about 15 °C, and then hold the mixture at that temperature for a time longer than about 1 minute; (h)Isolate the solid from the resulting mixture from step (g) to obtain Form I of the tris salt of 2 - [((4 - {6 - [((4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [((2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid; A method comprising the above steps.

12. A method for preparing Form 1 of the tris salt of 2 - [((4 - {6 - [((4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [((2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid, comprising: (a)Suspend the tris salt of 2 - [((4 - {6 - [((4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [((2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid in a solvent system, wherein the solvent system consists of tetrahydrofuran (THF) and water, the volume ratio of THF:water is from about 1:1 to about 4:1, and the ratio of water (volume) to the tris salt of 2 - [((4 - {6 - [((4 - cyano - 2 - fluorobenzyl)oxy]pyridin - 2 - yl}piperidin - 1 - yl)methyl] - 1 - [((2S)-oxetan - 2 - ylmethyl] - 1H - benzimidazole - 6 - carboxylic acid (weight) is from about 1.1 mL / g to about 3.8 mL / g at a temperature from about 20 °C to about 35 °C; (b)Heat the suspension in step (a) to a high temperature from about 49 °C to about 59 °C to form a solution, cool the temperature to a holding temperature from about 47 °C to about 51 °C, during which the mixture remains a solution, and optionally, mix the solution at the holding temperature for a time longer than about 1 minute; (c) To the solution from step (b), crystalline form 1 material of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid is seeded while maintaining the temperature at the holding temperature, where the amount of the seed crystal crystallinity is at least about 0.5% by weight of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid used in step (a), and then the resulting mixture is held at the holding temperature for a time longer than about 1 minute, and (d) Slowly cooling the temperature of the mixture from step (c) to an intermediate temperature of about 35 °C, and then holding the mixture at the intermediate temperature for a time longer than about 1 minute, and (e) Slowly adding a water-miscible organic solvent to the mixture from step (d) while maintaining the temperature of the mixture at the intermediate temperature, and then holding the mixture at the intermediate temperature for a time longer than about 1 minute, and (f) Slowly cooling the temperature of the mixture from step (e) to a low temperature of about 10 °C, and then holding the mixture at the low temperature for a time longer than about 1 minute, and (g) Sampling a sample of the mixture (slurry) to determine the particle size of the solid in the mixture, and (h) Performing high-shear wet milling until the D90 of the particle size of the solid in the mixture is less than about 150 μM, and (i) Isolating the solid from the resulting mixture from step (f) to obtain form I of the tris salt of 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid and A method comprising.

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