GLP-1 receptor agonists and uses thereof

Abstract

Provided herein are 6-formic acids of benzimidazoles and 4-aza-, 5-aza-, and 7-aza-benzimidazoles that are GLP-1R agonists, methods of making the compounds, and methods of administering the compounds to mammals in need thereof. # imgabs0 #

Description

[0001] This application is a divisional application of the patent application with application number 201980053632.5, application date June 11, 2019, and invention name “GLP-1 receptor agonists and their uses”. Technical Field

[0002] Provided herein are benzimidazoles and 6-carboxylic acids of 4-aza-, 5-aza-, and 7-aza-benzimidazoles as GLP-1R agonists, methods of making the compounds, and methods comprising administering the compounds to a mammal in need thereof. Background Art

[0003] Diabetes is an important public health problem due to its increasing prevalence and associated health risks. This disease is characterized by high levels of blood sugar caused by defects in insulin production, insulin action, or both. Two major forms of diabetes have been identified, type 1 and type 2. Type 1 diabetes (T1D) develops when the body's immune system destroys pancreatic β cells, which are the only cells in the body that make the hormone insulin (regulate blood sugar). In order to survive, people with type 1 diabetes must administer insulin by injection or pump. Type 2 diabetes (commonly known as T2DM) usually begins when insulin resistance occurs or when insulin is not fully produced to maintain acceptable blood sugar levels.

[0004] Various pharmacological approaches are currently available for the treatment of hyperglycemia and, consequently, T2DM (Hampp, C. et al. Use of Antidiabetic Drugs in the US, 2003-2012, Diabetes Care 2014, 37, 1367-1374). These can be grouped into six major categories, each acting through a different primary mechanism: (A) insulin-releasing drugs, including sulfonylureas (e.g., glipizide, glimepiride, glyburide), meglitinides (e.g., nateglidine, repaglinide), dipeptidyl peptidase IV (DPP-IV) inhibitors (e.g., sitagliptin, vildagliptin, agliflozin), and insulin-releasing drugs. (alogliptin, dutogliptin, linagliptin, saxogliptin), and glucagon-like peptide-1 receptor (GLP-1R) agonists (e.g., liraglutide, albiglutide, exenatide, lixisenatide, dulaglutide, semaglutide), which enhance insulin secretion by acting on pancreatic beta cells. Sulfonyl-ureas and meglitinides have limited efficacy and tolerability, cause weight gain and often induce hypoglycemia. DPP-IV inhibitors have limited efficacy. Marketed GLP-1R agonists are peptides administered by subcutaneous injection. Liraglutide is also approved for the treatment of obesity. (B) Biguanides (e.g., metformin) are believed to work primarily by reducing hepatic glucose production. Biguanides often cause gastrointestinal disturbances and lactic acidosis, further limiting their use. (C) Inhibitors of alpha-glucosidase (eg, acarbose) reduce intestinal glucose absorption. These agents often cause gastrointestinal disturbances. (D) Thiazolidinediones [eg, pioglitazone, rosiglitazone] act on specific receptors (peroxisome proliferator-activated receptor-gamma) in the liver, muscle, and adipose tissue. They modulate lipid metabolism and thereby enhance the responsiveness of these tissues to the action of insulin. Regular use of these drugs may lead to weight gain and may induce edema and anemia.(E) Insulin is used alone or in combination with the aforementioned agents for more severe cases, and regular use may also lead to weight gain and the risk of hypoglycemia. (F) Sodium-glucose linked transporter cotransporter 2 (SGLT2) inhibitors (e.g., dapagliflozin, empagliflozin, canagliflozin, ertugliflozin) inhibit the reabsorption of glucose in the kidneys and thus lower glucose levels in the blood. This emerging class of drugs may be associated with ketoacidosis and urinary tract infections.

[0005] However, with the exception of GLP-1R agonists and SGLT2 inhibitors, these drugs have limited efficacy and fail to address the most important issue, β-cell dysfunction and associated obesity.

[0006] In modern society, obesity is a very common chronic disease and is associated with many medical problems, including hypertension, hypercholesterolemia and coronary heart disease. It is also further highly associated with T2DM and insulin resistance, the latter usually accompanied by hyperinsulinemia or hyperglycemia, or both. In addition, T2DM is related to a two to fourfold increase in the risk of coronary artery disease. At present, the only treatment that can efficiently eliminate obesity is bariatric surgery, but this treatment is expensive and risky. Pharmacological interventions are usually less effective and involve side effects. Therefore, it is clearly necessary to have a more effective pharmacological intervention with fewer side effects and convenient administration.

[0007] Although T2DM is most commonly associated with hyperglycemia and insulin resistance, 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).

[0008] NAFLD is the hepatic manifestation of metabolic syndrome and is a spectrum of liver conditions that includes steatosis, nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and ultimately hepatocellular carcinoma. NAFLD and NASH are considered the main 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 extent of fibrosis. Although not all individuals with steatosis will progress to NASH, a significant portion of individuals will.

[0009] GLP-1 is a 30 amino acid long incretin hormone secreted by L-cells in the intestine in response to food ingestion. GLP-1 has been shown to stimulate insulin secretion in a physiological and glucose-dependent manner, reduce glucagon secretion, inhibit gastric emptying, reduce appetite, and stimulate β-cell proliferation. In nonclinical experiments, GLP-1 promotes sustained β-cell capacity by stimulating the transcription of genes important for glucose-dependent insulin secretion and by promoting β-cell neogenesis (Meier et al., Biodrugs. 2003; 17(2): 93-102).

[0010] In healthy individuals, GLP-1 plays an important role in regulating postprandial blood glucose levels by stimulating glucose-dependent insulin secretion from the pancreas, leading to increased peripheral glucose absorption. GLP-1 also suppresses glucagon secretion, leading to reduced glucose production in the liver. In addition, GLP-1 delays gastric emptying and slows small intestinal motility, thereby delaying food absorption. In people with T2DM, the normal postprandial rise of GLP-1 is absent or reduced (Vilsboll T et al., Diabetes. 2001. 50; 609-613).

[0011] Holst (Physiol. Rev. 2007, 87, 1409) and Meier (Nat. Rev. Endocrinol. 2012, 8, 728) described that GLP-1 receptor agonists, such as GLP-1, liraglutide and exendin-4, have three main pharmacological activities that improve glycemic control in patients with T2DM by lowering fasting and postprandial glucose (FPG and PPG): (i) increasing glucose-dependent insulin secretion (improved first and second phases), (ii) glucagon suppressive activity under hyperglycemic conditions, and (iii) delaying the rate of gastric emptying, thereby slowing the absorption of meal-derived glucose.

[0012] There remains a need for easily administered prevention and / or treatments of cardiometabolic and related diseases. Summary of the invention

[0013] The present invention relates to compounds of formula I

[0014]

[0015] or a pharmaceutically acceptable salt thereof, wherein

[0016] R is F, Cl, or -CN;

[0017] p is 0 or 1;

[0018] Ring A is phenyl or 6-membered heteroaryl;

[0019] m is 0, 1, 2, or 3;

[0020] Each R 1 Independently selected from: halogen, -CN, -C 1-3 Alkyl, and -OC 1-3 Alkyl, where C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group is substituted with 0 to 3 F atoms;

[0021] R 2 Is H or -C 1-3 Alkyl, wherein the alkyl group is substituted with 0 to 1 OH groups;

[0022] Each R 3 are independently F, -OH, -CN, -C 1-3 Alkyl, -OC 1-3 Alkyl, and -C 3-4 Cycloalkyl, or 2 R 3 Can cyclize together to form -C 3-4 Spirocycloalkyl, where, if valence permits, C 1-3 Alkyl and OC 1-3 The alkyl, cycloalkyl, or spirocycloalkyl group of the alkyl group may be substituted by 0 to 3 F atoms and by 0 to 1 -OH groups;

[0023] q is 0, 1, or 2;

[0024] XL is N-CH 2 、CHCH 2 , or cyclopropyl;

[0025] Y is CH or N;

[0026] R 4 Yes-C 1-3 Alkyl, -C 0-3 Alkylene-C 3-6 Cycloalkyl, -C 0-3 Alkylene-R 5 , or -C 1-3 Alkylene-R 6 , wherein if valence permits, the alkyl group may be substituted by 0 to 3 substituents independently selected from 0 to 3 F atoms and 0 to 1 substituent selected from -C 0-1 Alkylene -CN, -C 0-1 Alkylene-OR O 、-SO 2 -N(R N ) 2 、-C(O)-N(R N ) 2 、-N(C=O)(R N ), and -N(RN ) 2 substituted with a substituent; and

[0027] Wherein, if valence permits, the alkylene and cycloalkyl groups may be independently substituted with 0 to 2 substituents independently selected from 0 to 2 F atoms and 0 to 1 substituent selected from -C 0-1 Alkylene -CN, -C 0-1 Alkylene-OR O , and -N(R N ) 2 Substituents are substituted;

[0028] R 5 is a 4- to 6-membered heterocycloalkyl group, wherein the heterocycloalkyl group may be substituted, if valence permits, with 0 to 2 substituents independently selected from:

[0029] 0 to 1 oxo (=O),

[0030] 0 to 1 -CN,

[0031] 0 to 2 F atoms, and

[0032] 0 to 2 independently selected from -C 1-3 Alkyl and -OC 1-3 Alkyl substituents, where valence permits, C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group may be substituted with 0 to 3 substituents independently selected from the following:

[0033] 0 to 3 F atoms,

[0034] 0 to 1 -CN, and

[0035] 0 to 1 -OR O ;

[0036] R 6 is a 5- to 6-membered heteroaryl group, wherein, if valence permits, the heteroaryl group may be substituted with 0 to 2 substituents independently selected from:

[0037] 0 to 2 halogens,

[0038] 0 to 1 selected from -OR O and -N(R N ) 2 The substituents, and

[0039] 0 to 2 -C 1-3 Alkyl, wherein if valence permits, the alkyl may be substituted with 0 to 3 substituents independently selected from:

[0040] 0 to 3 F atoms, and

[0041] 0 to 1 -OR O ;

[0042] Each R O are independently H, or -C 1-3 Alkyl, where C 1-3 The alkyl group may be substituted by 0 to 3 F atoms;

[0043] Each R N are independently H, or -C 1-3 alkyl;

[0044] Z 1 , Z 2 , and Z 3 Each is -CR Z ,or

[0045] Z 1 , Z 2 , and Z 3 One of them is N and the other two are -CR Z ;and

[0046] Each R Z are independently H, F, Cl, or -CH 3 .

[0047] Another embodiment relates to compounds of formula II

[0048]

[0049] or a pharmaceutically acceptable salt thereof, wherein

[0050] R is F;

[0051] p is 0 or 1;

[0052] Ring A is phenyl or pyridyl;

[0053] m is 0, 1, or 2;

[0054] Each R 1 Independently selected from: halogen, -CN, -C 1-3 Alkyl, and -OC 1-3 Alkyl, where C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group is substituted with 0 to 3 F atoms;

[0055] R 2 is H or CH 3 ;

[0056] XL is N-CH 2 , or cyclopropyl;

[0057] Y is CH or N;

[0058] Z 3 Yes-CR Z or N; and

[0059] R Z is H, F, Cl, or -CH 3 .

[0060] Another embodiment relates to compounds of formula III

[0061]

[0062] or a pharmaceutically acceptable salt thereof, wherein

[0063] Ring A is phenyl or pyridyl;

[0064] m is 0, 1, or 2;

[0065] Each R 1 independently selected from F, Cl, and -CN;

[0066] R 2 is H or CH 3 ;and

[0067] Y is CH or N.

[0068] Another embodiment relates to compounds of formula IV

[0069]

[0070] or a pharmaceutically acceptable salt thereof, wherein

[0071] m is 0, 1, or 2;

[0072] Each R 1 independently selected from F, Cl, and -CN;

[0073] R 2 is H or CH 3 ;and

[0074] Y is CH or N.

[0075] Another embodiment relates to compounds of formula V

[0076]

[0077] or a pharmaceutically acceptable salt thereof, wherein

[0078] m is 0 or 1;

[0079] R 1 is F, Cl, or -CN;

[0080] R2 is H or CH 3 ;and

[0081] Y is CH or N.

[0082] Another embodiment relates to compounds of formula IV or formula V, wherein the phenyl or pyridyl group of ring A has one R 1 , which is substituted at the para position relative to the carbon at which the phenyl or pyridyl group is attached to the dioxolane to provide:

[0083]

[0084] or a pharmaceutically acceptable salt thereof, wherein

[0085] Each R 1 independently selected from F, Cl, and -CN;

[0086] R 2 is H or CH 3 ;and

[0087] Y is CH or N.

[0088] Another embodiment is directed to a compound of the other embodiments herein, e.g., a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, wherein XL is N—CH 2 ; and Y is CH or N. According to the embodiments described herein, in this case, X is N and L is CH 2 .

[0089] Another embodiment is directed to a compound of the other embodiments herein, e.g., a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, wherein XL is CHCH 2 ; and Y is N. According to the embodiments described herein, in this case, X is CH and L is CH 2 .

[0090] Another embodiment is directed to a compound of the other embodiments herein, e.g., a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, wherein XL is CHCH 2 ; and Y is CH. According to the embodiments described herein, in this case, X is CH and L is CH 2 .

[0091] Another embodiment is directed to a compound of the other embodiments herein, for example, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, wherein XL is cyclopropyl; and Y is N.

[0092] In embodiments where XL is cyclopropyl, the compound of Formula I or II provides:

[0093]

[0094] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein R 4 Yes-CH 2 CH 2 OCH 3 , C 1-3 Alkylene-R 5 , or C 1-3 Alkylene-R 6 .

[0095] Another embodiment is directed to a compound of Formula II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein R 4 As defined for compounds of formula I.

[0096] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein R 4 Yes-C 1-3 Alkyl, wherein if valence permits, the alkyl may be substituted with 0 to 1 substituents selected from: -C 0-1 Alkylene-OR O , and -N(R N ) 2 .

[0097] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein R 4 Yes-(CH 2 ) 2 OCH 3 , or -(CH 2 ) 2 N(CH 3 ) 2 .

[0098] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein R 4 Yes-CH 2 -R 5 , where R 5 is a 4- to 5-membered heterocycloalkyl group, wherein, if valence permits, the heterocycloalkyl group may be substituted with 0 to 2 substituents independently selected from:

[0099] 0 to 2 F atoms, and

[0100] 0 to 1 selected from -OCH 3 and -CH 2 OCH 3substituent.

[0101] Another embodiment is directed to compounds of Formula I, II, III, IV, or V, wherein the heterocycloalkyl group is

[0102]

[0103] Where valence permits, the heterocycloalkyl group may be substituted with 0 to 2 substituents independently selected from the following (e.g., replacing hydrogen):

[0104] 0 to 1 oxo (O=),

[0105] 0 to 1 -CN,

[0106] 0 to 2 F atoms, and

[0107] 0 to 2 independently selected from -C 1-3 Alkyl and -OC 1-3 Alkyl substituents, where valence permits, C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group may be independently substituted with 0 to 3 substituents independently selected from:

[0108] 0 to 3 F atoms,

[0109] 0 to 1 -CN, and

[0110] 0 to 1 -OR O ,

[0111] or a pharmaceutically acceptable salt thereof.

[0112] Another embodiment is directed to compounds of Formula I, II, III, IV, or V, wherein the heterocycloalkyl group is

[0113]

[0114] Where valence permits, the heterocycloalkyl group may be substituted with 0 to 2 substituents independently selected from the following (e.g., replacing hydrogen):

[0115] 0 to 1 -CN,

[0116] 0 to 2 F atoms, and

[0117] 0 to 2 independently selected from -C 1-3 Alkyl and -OC 1-3 Alkyl substituents, where valence permits, C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group may be independently substituted with 0 to 3 substituents independently selected from:

[0118] 0 to 3 F atoms,

[0119] 0 to 1 -CN, and

[0120] 0 to 1 -OR O ,

[0121] or a pharmaceutically acceptable salt thereof.

[0122] Another embodiment is directed to compounds of Formula I, II, III, IV, or V, wherein the heterocycloalkyl group is

[0123]

[0124] Where valence permits, the heterocycloalkyl group may be substituted with 0 to 1 substituents selected from the following (e.g., replacing hydrogen):

[0125] -CN,

[0126] F atoms, and

[0127] 0 to 1 independently selected from -C 1-3 Alkyl and -OC 1-3 Alkyl substituents, where valence permits, C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group may be substituted with 0 to 3 substituents independently selected from the following:

[0128] 0 to 3 F atoms,

[0129] 0 to 1 -CN, and

[0130] 0 to 1 -OR O ,

[0131] or a pharmaceutically acceptable salt thereof.

[0132] Another embodiment is directed to compounds of Formula I, II, III, IV, or V, wherein the heterocycloalkyl group is

[0133]

[0134] or a pharmaceutically acceptable salt thereof.

[0135] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein the heterocycloalkyl group is

[0136]

[0137] Where valence permits, the heterocycloalkyl group may be substituted with 0 to 1 methyl groups, wherein the methyl groups may be substituted with 0 to 3 F atoms.

[0138] Another embodiment is directed to compounds of Formula I, II, III, IV, or V, wherein the heterocycloalkyl group is

[0139]

[0140] wherein the heterocycloalkyl group is unsubstituted.

[0141] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein -CH 2 -R 5 With R 4 Connected nitrogen provides:

[0142]

[0143] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein R 4 Yes-CH 2 -R 6 , where R 6 is a 5-membered heteroaryl, wherein, if valence permits, the heteroaryl may be substituted with 0 to 2 substituents independently selected from:

[0144] 0 to 2 halogens, wherein halogens are independently selected from F and Cl,

[0145] 0 to 1 -OCH 3 ,and

[0146] 0 to 1 -CH 3 、-CH 2 CH 3 , -CF 3 , or -CH 2 CH 2 OCH 3 .

[0147] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein the heteroaryl group is

[0148]

[0149] Wherein, if valence permits, the heteroaryl group may be substituted with 0 to 2 substituents independently selected from the following (e.g., replacing hydrogen):

[0150] 0 to 2 halogens, wherein halogens are independently selected from F and Cl,

[0151] 0 to 1 selected from -OR O and -N(R N ) 2 A substituent, or

[0152] 0 to 2 -C 1-3 Alkyl, wherein if valence permits, the alkyl may be substituted with 0 to 3 substituents independently selected from:

[0153] 0 to 3 F atoms, and

[0154] 0 to 1 -OR O .

[0155] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein the heteroaryl group is

[0156]

[0157] Wherein, if valence permits, the heteroaryl group may be substituted with 0 to 2 substituents independently selected from the following (e.g., replacing hydrogen):

[0158] 0 to 2 halogens, wherein halogens are independently selected from F and Cl,

[0159] 0 to 1 selected from -OR O and -N(R N ) 2 or

[0160] 0 to 2 -C 1-3 Alkyl, wherein if valence permits, the alkyl may be substituted with 0 to 3 substituents independently selected from:

[0161] 0 to 3 F atoms, and

[0162] 0 to 1 -OR O .

[0163] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein the heteroaryl group is

[0164]

[0165] Wherein, if valence permits, the heteroaryl group may be substituted with 0 to 1 -C 1-2 Alkyl substituents, wherein if valence permits, the alkyl group may be substituted with 0 to 3 substituents independently selected from:

[0166] 0 to 3 F atoms, and

[0167] 0 to 1 -OR O ;and

[0168] Each R O are independently H, or -C 1-3 alkyl.

[0169] It is recognized that any substituent may replace the H on the substituted carbon or nitrogen. Non-limiting examples of substituted heteroaryl groups are:

[0170]

[0171] It is recognized that H is substituted by, for example, R 6s (R 6 ) to provide:

[0172]

[0173] Where R 6s Yes-C 1-2 Alkyl, wherein if valence permits, the alkyl may be substituted with 0 to 3 substituents independently selected from:

[0174] 0 to 3 F atoms, and

[0175] 0 to 1 -R O ;and

[0176] Each R O are independently H, or -C 1-3 alkyl;

[0177] or a pharmaceutically acceptable salt thereof.

[0178] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein heteroaryl is:

[0179]

[0180] Another embodiment is directed to a compound of the other embodiments herein, or a pharmaceutically acceptable salt thereof, for example, a compound of Formula I, II, III, IV, or V, wherein Z 1 , Z 2 , and Z 3 Each is CR Z .

[0181] Another embodiment is directed to a compound of the other embodiments herein, or a pharmaceutically acceptable salt thereof, for example, a compound of Formula I, II, III, IV, or V, wherein R Z It's H.

[0182] Another embodiment is directed to a compound of the other embodiments herein, or a pharmaceutically acceptable salt thereof, for example, a compound of Formula I, II, III, IV, or V, wherein Z 1 , Z 2 , and Z 3Each is CH.

[0183] Another embodiment is directed to a compound of the other embodiments herein, or a pharmaceutically acceptable salt thereof, for example, a compound of Formula I, II, III, IV, or V, wherein R 3 Yes-CH 3 , or -CF 3 ; and q is 1.

[0184] Another embodiment is directed to a compound of the other embodiments herein, or a pharmaceutically acceptable salt thereof, e.g., a compound of Formula I, II, III, IV, or V, wherein each R 1 are independently F, Cl, or -CN.

[0185] Another embodiment is directed to a compound of the other embodiments herein, or a pharmaceutically acceptable salt thereof, for example, a compound of Formula I, II, III, IV, or V, wherein R 4 Yes-CH 2 -R 5 .

[0186] Another embodiment is directed to a compound of the other embodiments herein, or a pharmaceutically acceptable salt thereof, for example, a compound of Formula I, II, III, IV, or V, wherein R 4 Yes-CH 2 -R 6 .

[0187] Another embodiment is directed to a compound of the other embodiments herein, for example, a compound of Formula I, II, III, IV, or V, wherein the compound is a free acid.

[0188] Another embodiment is directed to any embodiment of a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein Ring A and R 2 supply:

[0189]

[0190] in:

[0191] R is F, Cl, or -CN;

[0192] p is 0 or 1;

[0193] m is 0, 1, or 2; and

[0194] Each R 1 Independently selected from: halogen, -CN, -C 1-3 Alkyl, and -OC 1-3 Alkyl, where C 1-3 Alkyl and OC 1-3The alkyl group of the alkyl group is substituted with 0 to 3 F atoms.

[0195] Another embodiment is directed to a compound of Formula I, II, III, IV, or V, or a pharmaceutically acceptable salt thereof, wherein R 2 It's H.

[0196] Another embodiment relates to wherein R 2 is a compound of the present invention wherein H, or a pharmaceutically acceptable salt thereof.

[0197] Another embodiment relates to compounds of the present invention, wherein said compound is

[0198] 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or

[0199] 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0200] or a pharmaceutically acceptable salt thereof.

[0201] Another embodiment relates to compounds of the present invention, wherein said compound is

[0202] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or

[0203] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0204] or a pharmaceutically acceptable salt thereof.

[0205] Another embodiment relates to wherein R 2 Yes CH 3 The compound of the present invention, or a pharmaceutically acceptable salt thereof.

[0206] Another embodiment relates to compounds of the present invention, wherein said compound is

[0207] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0208] 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0209] 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0210] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-3-(1,3-oxazol-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid;

[0211] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0212] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-4-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0213] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(pyridin-3-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0214] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-5-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0215] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-1,2,3-triazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0216] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0217] 2-({4-[2-(4-chloro-2-fluorophenyl)-7-fluoro-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0218] 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylic acid; or

[0219] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0220] or a pharmaceutically acceptable salt thereof.

[0221] Another embodiment relates to compounds of the present invention, wherein said compound is

[0222] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or

[0223] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0224] or a pharmaceutically acceptable salt thereof.

[0225] Another embodiment relates to compounds of the present invention, wherein said compound is

[0226] 2-({4-[(2S)-2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0227] 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or

[0228] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0229] or a pharmaceutically acceptable salt thereof.

[0230] Another embodiment relates to compounds of the present invention, wherein said compound is

[0231] 2-({4-[(2R)-2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0232] 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or

[0233] 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0234] or a pharmaceutically acceptable salt thereof.

[0235] Another embodiment relates to compounds of the present invention, wherein said compound is

[0236] 2-({4-[2-(4-cyano-2-fluorophenyl)-2*-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, wherein the chirality of 2* comes from C56;

[0237] 2-({4-[2-(5-chloropyridin-2-yl)-2*-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, wherein the chirality of 2* comes from P9;

[0238] 2-({4-[2-(4-chloro-2-fluorophenyl)-2*-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid, wherein the chirality of 2* comes from 17;

[0239] 2-({4-[2-(4-chloro-2-fluorophenyl)-7-fluoro-2*-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, wherein the chirality of 2* is derived from 96; or

[0240] 2-({4-[2-(4-cyano-2-fluorophenyl)-2*-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylic acid, wherein the chirality of 2* comes from C82;

[0241] or a pharmaceutically acceptable salt thereof.

[0242] Another embodiment includes a compound which is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, or a pharmaceutically acceptable salt thereof, wherein the salt is a tris salt.

[0243] Another embodiment includes the compound which is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid as the free acid.

[0244] Another embodiment includes compounds which are

[0245]

[0246] or a pharmaceutically acceptable salt thereof.

[0247] Another embodiment includes the compound 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, or a pharmaceutically acceptable salt thereof, wherein the salt is a tris salt {the tris salt of this compound is also known as: 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate}.

[0248] In certain embodiments, the present invention provides a crystalline form of anhydrous tris salt of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid. In certain further embodiments, the crystalline form of the anhydrous (anhydrate) tris salt of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid is designated as "Form I" based on its unique solid-state characteristics with respect to, for example, powder X-ray diffraction (PXRD) as described herein (e.g., substantially as described herein). Figure 1In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising at least two characteristic peaks selected from 3.7 ± 0.2°; 7.3 ± 0.2°; 8.5 ± 0.2°; 10.1 ± 0.2°; 14.7 ± 0.2°; and 16.9 ± 0.2°. In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising at least three characteristic peaks selected from 3.7 ± 0.2°; 7.3 ± 0.2°; 8.5 ± 0.2°; 10.1 ± 0.2°; 14.7 ± 0.2°; and 16.9 ± 0.2°. In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising at least four characteristic peaks selected from 3.7 ± 0.2°; 7.3 ± 0.2°; 8.5 ± 0.2°; 10.1 ± 0.2°; 14.7 ± 0.2°; and 16.9 ± 0.2°. In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising at least five characteristic peaks selected from 3.7 ± 0.2°; 7.3 ± 0.2°; 8.5 ± 0.2°; 10.1 ± 0.2°; 14.7 ± 0.2°; and 16.9 ± 0.2°.

[0249] In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising characteristic peaks at 3.7±0.2° and 7.3±0.2° 2θ.

[0250] In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising peaks at 3.7±0.2°, 7.3±0.2°, and 14.7±0.2°, 2θ. In certain further embodiments, Form I exhibits a powder X-ray diffraction pattern further comprising at least one peak selected from 8.5±0.2°, 10.1±0.2°, and 16.9±0.2°, 2θ.

[0251] In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising peaks at 3.7±0.2°; 7.3±0.2°; 14.7±0.2°; and 16.9±0.2° 2θ.

[0252] In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising peaks at 3.7±0.2°; 7.3±0.2°; 8.5±0.2°; 10.1±0.2°; 14.7±0.2°; and 16.9±0.2°, with respect to 2θ.

[0253] In certain embodiments, Form I exhibits substantially Figure 1The powder X-ray diffraction pattern shown is shown in Table X1 above. A series of diffraction peaks expressed in terms of 2θ degrees and relative intensity (relative intensity ≥ 3.0%) are provided.

[0254] As is well known in the art of powder diffraction, the relative intensities of peaks (reflections) may vary with sample preparation techniques, sample installation procedures, and the specific instrument used. In addition, instrument variation and other factors may affect 2-θ values. Therefore, XRPD peak attributions may vary by ± about 0.2°.

[0255] Another embodiment includes the compound which is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid as the free acid.

[0256] Another embodiment includes compounds which are

[0257]

[0258] or a pharmaceutically acceptable salt thereof.

[0259] Another embodiment includes compounds which are the following compounds as free acids:

[0260] 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0261] 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or

[0262] 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid.

[0263] Another embodiment includes compounds: which are

[0264] 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0265] 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or

[0266] 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0267] or a pharmaceutically acceptable salt thereof, wherein the salt is a tris salt.

[0268] Another embodiment includes the compound which is 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2:

[0269]

[0270] or a pharmaceutically acceptable salt thereof. In certain further embodiments, the present invention provides a compound which is 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2, or a tris salt thereof [i.e., 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine salt]. The chiral center on the left portion of the compound structure is labeled "abs" to indicate that the chiral center has only one stereo configuration (i.e., not a racemic form relative to the chiral center).

[0271] In certain embodiments, the present invention provides a crystalline form of the anhydrous tris salt of 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2. In certain further embodiments, the crystalline form of the anhydrous (anhydrate) tris salt of 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2, is designated as "Form A" based on its unique solid-state characteristics with respect to, for example, powder X-ray diffraction (PXRD) as described herein (e.g., substantially as described herein). Figure 2 Depicted). In certain embodiments, Form A exhibits a powder X-ray diffraction pattern comprising at least two characteristic peaks selected from 7.7±0.2°; 15.2±0.2°; 15.7±0.2° and 17.6±0.2° in terms of 2θ. In certain embodiments, Form A exhibits a powder X-ray diffraction pattern comprising at least three characteristic peaks selected from 7.7±0.2°; 15.2±0.2°; 15.7±0.2° and 17.6±0.2° in terms of 2θ. In certain embodiments, Form A exhibits a powder X-ray diffraction pattern comprising characteristic peaks selected from 7.7±0.2°; 15.2±0.2°; 15.7±0.2° and 17.6±0.2° in terms of 2θ.

[0272] In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising characteristic peaks at 7.7±0.2°; and 17.6±0.2° 2θ.

[0273] In certain embodiments, Form A exhibits a powder X-ray diffraction pattern comprising peaks at 7.7±0.2°; 15.2±0.2°; and 17.6±0.2° 2θ.

[0274] In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising peaks at 7.7±0.2°; 15.2±0.2°; and 15.7±0.2° 2θ.

[0275] In certain embodiments, Form I exhibits a powder X-ray diffraction pattern comprising peaks at 7.7±0.2°; 15.2±0.2°; 15.7±0.2° and 17.6±0.2° 2θ.

[0276] In certain embodiments, Form A exhibits substantially Figure 2The powder X-ray diffraction pattern shown is shown in Table X2 above. A series of diffraction peaks expressed in terms of 2θ degrees and relative intensity (relative intensity ≥ 3.0%) are provided.

[0277] As is well known in the art of powder diffraction, the relative intensities of peaks (reflections) may vary with sample preparation techniques, sample installation procedures, and the specific instrument used. In addition, instrument variation and other factors may affect 2-θ values. Therefore, XRPD peak attributions may vary by ± about 0.2°.

[0278] In another embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, in admixture with at least one pharmaceutically acceptable excipient. This includes a pharmaceutical composition comprising a compound of formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, in admixture with at least one pharmaceutically acceptable excipient and one or more other therapeutic agents discussed herein.

[0279] The present invention also includes the following embodiments:

[0280] A compound of formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, for use as a medicament;

[0281] A compound of Formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, for use in preventing and / or treating cardiometabolic and related diseases discussed herein, including T2DM, prediabetes, NASH, and cardiovascular disease;

[0282] A method of treating a GLP-1R agonist-indicated disease in a subject in need of such prevention and / or treatment, comprising administering to the subject a therapeutically effective amount of a compound of Formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof;

[0283] Use of a compound of formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease or condition for which a GLP-1R agonist is indicated;

[0284] A compound of formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, for use in treating a disease or condition for which a GLP-1R agonist is indicated; or

[0285] A pharmaceutical composition for treating a disease or condition for which a GLP-1R agonist is indicated, comprising a compound of Formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof.

[0286] Each example or a pharmaceutically acceptable salt thereof may be claimed individually, or grouped together with any number of each of the embodiments described herein in any combination.

[0287] The present invention also relates to a pharmaceutical composition comprising a compound of Formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment and / or prevention of the cardiovascular metabolic and related diseases discussed herein (including T2DM, prediabetes, NASH, and cardiovascular disease).

[0288] Another embodiment of the invention relates to a compound of formula I, II, III, IV, or V as defined in any of the embodiments described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment and / or prevention of cardiovascular metabolic and related diseases including: diabetes (T1D and / or T2DM, including prediabetes), idiopathic T1D (type 1b), latent autoimmune diabetes in adults (LADA), early-onset T2DM (EOD), atypical diabetes of the juvenile onset (YOAD), maturity-onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, hyperglycemia , insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, renal disease (e.g., acute renal dysfunction, tubular dysfunction, proinflammatory changes in the proximal tubules), diabetic retinopathy, adipocyte dysfunction, visceral fat deposition, sleep apnea, obesity (including hypothalamic obesity and monogenic obesity) and related comorbidities (e.g., osteoarthritis and urinary incontinence), eating disorders (including binge eating disorder, bulimia nervosa, and syndromic obesity, such as Prader-Willi and Bardet-Biedl syndromes), obesity due to the use of other Weight gain due to medications (e.g., caused by the use of steroids and antipsychotics), excessive sugar cravings, dyslipidemia (including hyperlipidemia, hypertriglyceridemia, increased total cholesterol, high LDL cholesterol, and low HDL cholesterol), hyperinsulinemia, NAFLD (including related diseases, such as steatosis, NASH, fibrosis, cirrhosis, and hepatocellular carcinoma), cardiovascular disease, atherosclerosis (including coronary artery disease), peripheral vascular disease, hypertension, endothelial dysfunction, impaired vascular compliance, congestive heart failure, myocardial infarction (e.g., necrosis and apoptosis), stroke, hemorrhagic Stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidosis, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, macular degeneration, cataract, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina, thrombosis, atherosclerosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, impaired fasting glucose, hyperuricemia, gout, erectile dysfunction, skin and connective tissue disorders, psoriasis, foot ulcers, ulcerative colitis, high apo Prevention or treatment of hyper apo Blipoproteinemia, Alzheimer's disease, schizophrenia, impaired cognitive function, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel syndrome, polycystic ovary syndrome, and treatment of addiction (e.g., alcohol and / or drug abuse).

[0289] The abbreviations used in this article are as follows:

[0290] The term "alkyl" as used herein refers to a n H (2n+1) Non-limiting examples include methyl, ethyl, propyl, butyl, 2-methyl-propyl, 1,1-dimethylethyl, pentyl, and hexyl.

[0291] The term "alkylene" as used herein refers to a n H 2n - is a straight or branched divalent hydrocarbon group. Non-limiting examples include ethylene and propylene.

[0292] The term "cycloalkyl" as used herein refers to a radical of the formula -C n H (2n-1) Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0293] As used herein, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0294] The term "heterocycloalkyl" as used herein refers to a cycloalkyl group in which one or more cyclomethylene groups (-CH 2 -) has been selected from -O-, -S- or nitrogen-substituted cycloalkyl groups, wherein the nitrogen can provide the point of attachment or can be substituted as provided in each embodiment. In the case where nitrogen provides the point of attachment, the structure diagram of the heterocycloalkyl will have a hydrogen on the nitrogen. In general, if valence permits, the heterocycloalkyl can be substituted by 0 to 2 substituents independently selected from the following: oxo, -CN, halogen, alkyl and -O alkyl, and the alkyl can be further substituted. It can be noted that when there are 0 substitutions, the heterocycloalkyl is unsubstituted.

[0295] The term "heteroaryl" as used herein refers to a monocyclic aromatic hydrocarbon containing 5 to 6 carbon atoms, wherein at least one ring carbon atom is replaced by a heteroatom selected from oxygen, nitrogen and sulfur. Such heteroaryl groups may be attached via a ring carbon atom or, where valence permits, via a ring nitrogen atom. In general, heteroaryl groups may be substituted, where valence permits, by 0 to 2 substituents independently selected from the group consisting of halogen, OH, alkyl, O-alkyl, and amino (e.g., NH 2 、NH alkyl、N(alkyl) 2 ), and the alkyl group may be further substituted. It may be noted that when there are 0 substitutions, the heteroaryl group is unsubstituted.

[0296] Room temperature: RT (15 to 25°C).

[0297] Methanol: MeOH.

[0298] Ethanol: EtOH.

[0299] Isopropyl alcohol: iPrOH.

[0300] Ethyl acetate: EtOAc.

[0301] Tetrahydrofuran: THF.

[0302] Toluene: PhCH 3 .

[0303] Cesium carbonate: Cs 2 CO 3 .

[0304] Lithium bis(trimethylsilyl)amide: LiHMDS.

[0305] Sodium tert-butoxide: NaOtBu.

[0306] Potassium tert-butoxide: KOtBu.

[0307] Lithium diisopropylamide: LDA.

[0308] Triethylamine: Et 3 N.

[0309] N,N-Diisopropylethylamine: DIPEA.

[0310] Potassium carbonate: K 2 CO 3 .

[0311] Dimethylformamide: DMF.

[0312] Dimethylacetamide: DMAc.

[0313] Dimethyl sulfoxide: DMSO.

[0314] N-Methyl-2-pyrrolidone: NMP.

[0315] Sodium hydride: NaH.

[0316] Trifluoroacetic acid: TFA.

[0317] Trifluoroacetic anhydride: TFAA.

[0318] Acetic anhydride: Ac 2 O.

[0319] Dichloromethane: DCM.

[0320] 1,2-Dichloroethane: DCE.

[0321] Hydrochloric acid: HCl.

[0322] 1,8-diazabicyclo[5.4.0]undec-7-ene: DBU.

[0323] Borane-dimethyl sulfide complex: BH 3 -DMS.

[0324] Borane-tetrahydrofuran complex: BH 3 -THF.

[0325] Lithium aluminum hydride: LAH.

[0326] Acetic acid: AcOH.

[0327] Acetonitrile: MeCN.

[0328] p-Toluenesulfonic acid: pTSA.

[0329] Dibenzylideneacetone: DBA.

[0330] 2,2'-Bis(diphenylphosphino)-1,1'-binaphthyl: BINAP. 1,1'-Ferrocenyldiyl-bis(diphenylphosphine): dppf.

[0331] 1,3-Bis(diphenylphosphino)propane: DPPP.

[0332] 3-Chloroperoxybenzoic acid: m-CPBA.

[0333] Tert-butyl methyl ether: MTBE.

[0334] Methanesulfonyl: Ms.

[0335] N-Methylpyrrolidone: NMP.

[0336] Thin layer chromatography: TLC.

[0337] Supercritical fluid chromatography: SFC.

[0338] 4-(Dimethylamino)pyridine: DMAP.

[0339] tert-Butyloxycarbonyl: Boc.

[0340] 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate: HATU.

[0341] Petroleum ether: PE.

[0342] 2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate: HBTU.

[0343] 2-Amino-2-(hydroxymethyl)propane-1,3-diol: tris.

[0344] Tris(dibenzylideneacetone)dipalladium: Pd 2 (dba) 3 .

[0345] 1 H nuclear magnetic resonance (NMR) spectra were consistent with the proposed structures in all cases. Characteristic chemical shifts (δ) are expressed relative to the deuterated solvent (CHCl at 7.27 ppm). 3 ; CD at 3.31ppm 2 The residual proton signal is given in parts per million in HOD; MeCN at 1.94 ppm; DMSO at 2.50 ppm and is recorded using conventional abbreviations for designating major peaks: e.g., s, singlet; d, doublet; t: triplet; q, quartet; m, multiplet; br, broad. The symbol ^ indicates 1 The H NMR peak areas are assumed because the peaks are partially obscured by the water peak. 1 HNMR peak areas are assumed because the peaks are partially obscured by solvent peaks.

[0346] The wavy lines used in this article It indicates the point of attachment of a substituent to another group.

[0347] The compounds and intermediates described below are named using the naming conventions provided by ACD / ChemSketch 2012, File Version C10H41, Build 69045 (Advanced Chemistry Development, Inc., Toronto, Ontario, Canada). The naming conventions provided by ACD / ChemSketch 2012 are well known to those skilled in the art and the naming conventions provided by ACD / ChemSketch 2012 are considered to be generally consistent with the IUPAC (International Union for Pure and Applied Chemistry) recommendations and CAS indexing rules for organic chemical nomenclature. It can be noted that a chemical name may have only parentheses or may have parentheses and square brackets. Depending on the naming convention, stereochemical descriptors may also be placed in different positions within the name itself. Those skilled in the art will recognize these format variations and understand that they provide the same chemical structure.

[0348] Pharmaceutically acceptable salts of the compounds of Formula I, II, III, IV, or V include acid addition and base salts.

[0349] Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hyaluronate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, Maleate, malonate, methanesulfonate, methylsulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, toluenesulfonate, trifluoroacetate, 1,5-naphthalene disulfonate, and xinofoate salts.

[0350] Suitable base salts are formed from bases which form non-toxic salts. Examples include aluminum, arginine, benzathine penicillin, calcium, choline, diethylamine, bis(2-hydroxyethyl)amine (diethanolamine), glycine, lysine, magnesium, meglumine, 2-aminoethanol (ethanolamine), potassium, sodium, 2-amino-2-(hydroxymethyl)propane-1,3-diol (tris or tromethamine) and zinc salts.

[0351] Hemi-salts of acids and bases can also be formed, such as hemisulphate and hemicalcium salts. For a review of suitable salts, see Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (Wiley-VCH, 2002).

[0352] Pharmaceutically acceptable salts of compounds of formula I can be prepared by one or more of the following three methods:

[0353] (i) reacting a compound of formula I with a desired acid or base;

[0354] (ii) removing an acid- or base-labile protecting group from a suitable precursor of a compound of formula I, or ring-opening a suitable cyclic precursor (e.g., a lactone or lactam), using a desired acid or base; or

[0355] (iii) converting one salt of a compound of formula I into another salt by reaction with an appropriate acid or base or with the aid of a suitable ion exchange column.

[0356] All three reactions are usually carried out in solution. The resulting salt can be precipitated and collected by filtration, or can be recovered by evaporation of the solvent. The degree of ionization of the resulting salt can vary from fully ionized to almost non-ionized.

[0357] The compounds of formula I and their pharmaceutically acceptable salts can exist in unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of formula I or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol). When the solvent is water, the term "hydrate" is used.

[0358] The currently accepted classification system for organic hydrates is one that defines isolated site, channel or metal-ion coordinated hydrates - see KR Morris's Polymorphism in Pharmaceutical Solids (Editors HG Brittain, Marcel Dekker, 1995). Isolated site hydrates are hydrates in which the water molecules are separated from each other by intervening organic molecules, not in direct contact. In channel hydrates, the water molecules are located in lattice channels next to other water molecules. In metal-ion coordinated hydrates, the water molecules are bonded to the metal ions.

[0359] When the solvent or water is tightly bound, the complex may have a well-defined stoichiometry that is independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water / solvent content may depend on humidity and drying conditions. In such cases, non-stoichiometry will be the norm.

[0360] Also included within the scope of the present invention are multi-component complexes (except salts and solvates) in which the drug and at least one other component are present in stoichiometric or non-stoichiometric amounts. This type of complex includes clathrates (drug-host including complexes) and co-crystals. The latter is generally defined as a crystalline complex of neutral molecular components bound together via non-covalent interactions, but may also be a complex of a neutral molecule with a salt. Co-crystals can be prepared by melt crystallization, by recrystallization from a solvent, or by physically grinding the components together. See O. Almarsson and MJ Zaworotko, Chem Commun, 17 , 1889-1896 (2004). For a general review of multicomponent complexes, see Haleblian, JPharmSci, 64 (8), 1269-1288 (August 1975).

[0361] The compounds of the present invention can exist in a continuum of solid states ranging from completely amorphous to completely crystalline. The term "amorphous" refers to a state in which a material lacks long range order at the molecular level and can exhibit the physical properties of a solid or a liquid depending on the temperature. Such materials generally do not produce a unique X-ray diffraction pattern and, despite exhibiting solid properties, are more formally described as liquids. Upon heating, a change in solid to liquid properties occurs, characterized by a change in state, typically a secondary change ("glass transition"). The term "crystalline" refers to a solid phase in which a material has a regularly ordered internal structure at the molecular level and produces a unique X-ray diffraction pattern with well-defined peaks. Such materials will also exhibit the properties of a liquid when fully heated, but the change from solid to liquid is characterized by a phase transition, typically a primary phase transition ("melting point").

[0362] Compounds of Formula I may also exist in a mesomorphic state (mesophase or liquid crystal) when subjected to suitable conditions. The mesomorphic state is an intermediate state between a true crystalline state and a true liquid state (melt or solution). Mesomorphism due to temperature changes is described as "thermotropic", and mesomorphism due to the addition of a second component (such as water or another solvent) is described as "soluble". Compounds with the potential to form soluble mesophases are described as "amphoteric", and are composed of compounds with ionic polar head groups (such as -COO - Na + 、-COO - K + or -SO 3 - Na + ) or nonionic polar head groups (such as -N - N + (CH 3 ) 3 ) molecular composition. For more information, see NH Hartshorne and A. Stuart Crystals and the Polarizing Microscope , 4th edition (Edward Arnold, 1970).

[0363] The compounds of Formula I may exhibit polymorphism and / or one or more isomerisms (e.g., optical isomerism, geometric isomerism, or tautomerism). The compounds of Formula I may also be isotopically labeled. Such variations imply compounds of Formula I that are defined herein by reference to their structural features and are therefore within the scope of the present invention.

[0364] Compounds of formula I containing one or more asymmetric carbon atoms can exist as two or more stereoisomers. When compounds of formula I contain alkenyl or alkenylene, geometric cis / trans (or Z / E) isomers are possible. When structural isomers can be mutually converted via low energy barriers, tautomerism ("tautomerism") may exist. This can take the form of proton tautomerism in compounds of formula I containing, for example, imino, keto or oxime groups, or can take the form of so-called valence tautomerism in compounds containing aromatic moieties. As a result, a single compound can exhibit more than one type of isomerism.

[0365] Pharmaceutically acceptable salts of compounds of Formula I may also contain optically active (eg, d-lactate or 1-lysine) or racemic (eg, dl-tartrate or dl-arginine) counterions.

[0366] Cis / trans isomers may be separated by conventional techniques well known to those skilled in the art (eg chromatography and fractional crystallization).

[0367] Conventional techniques for preparing / separating individual enantiomers include chiral synthesis from suitable optically pure precursors or separation of racemates (or racemates of salts or derivatives) using, for example, chiral high pressure liquid chromatography (HPLC). Alternatively, racemic precursors containing chiral esters can be separated by enzymatic resolution (see, for example, Int J Mol Sci 29682-29716 (2015) by ACLM Carvaho et al.). In the case where the compound of Formula I contains an acidic or basic moiety, the salt can be formed using an optically pure base or acid (e.g., 1-phenylethylamine or tartaric acid). The resulting diastereomeric mixture can be separated by fractional crystallization and one or two diastereomeric salts can be converted into the corresponding pure enantiomers by means well known to technicians. Alternatively, the racemate (or racemic precursor) can be covalently reacted with an appropriate optically active compound (e.g., alcohol, amine or benzyl chloride). The resulting diastereomeric mixture can be separated by chromatography and / or fractional crystallization by means well known to the skilled person to obtain the separated diastereomers in the form of single enantiomers with 2 or more chiral centers. Using chromatography (usually HPLC) on an asymmetric resin using a mobile phase consisting of a hydrocarbon (usually heptane or hexane) containing 0 to 50% by volume (usually 2% to 20% by volume) of isopropanol and 0 to 5% by volume of an alkylamine (usually 0.1% diethylamine) ), chiral compounds of Formula I (and chiral precursors thereof) can be obtained in an enantiomerically enriched form. Concentration of the eluate can yield an enriched mixture. Chiral chromatography using sub- and supercritical fluids can be employed. Methods of chiral chromatography useful in certain embodiments of the present invention are known in the art (see, e.g., Smith, Roger M., Loughborough University, Loughborough, UK; Chromatographic Science Series (1998), 75 (SFC with Packed Columns), pp. 223-249 and references cited therein). In certain relevant examples herein, the columns are obtained from Chiral Technologies, Inc, West Chester, Pennsylvania, USA, A subsidiary of Chemical Industries, Ltd., Tokyo, Japan.

[0368] When any racemate is crystallized, two different types of crystals are possible. The first type is the racemic compound (real racemate) mentioned above, which wherein produces a homogeneous crystalline form containing two enantiomers in equimolar amounts. The second type is a racemic mixture or agglomerate, which wherein produces two crystalline forms of each comprising a single enantiomer in equimolar amounts. Although the two crystalline forms present in the racemic mixture may have the same physical properties, it 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, the Stereochemistry of Organic Compounds (Wiley, 1994) of ELEiel and SH Wilen.

[0369] It must be emphasized that, herein, the compound of Formula I is drawn in a single tautomeric form and that all possible tautomeric forms are included within the scope of the present invention.

[0370] The present invention includes all pharmaceutically acceptable isotopically-labeled compounds of formula I wherein one or more atoms are replaced by atoms having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature.

[0371] Examples of isotopes suitable for inclusion in the compounds of the present invention include: isotopes of hydrogen, for example, 2 H and 3 H; isotopes of carbon, e.g. 11 C. 13 C and 14 C; isotopes of chlorine, e.g. 36 Cl; isotopes of fluorine, e.g. 18 F; isotopes of iodine, e.g. 123 I and 125 I; isotopes of nitrogen, e.g. 13 N and 15 N; isotopes of oxygen, e.g. 15 Oh, 17 O and 18 O; isotopes of phosphorus, e.g. 32 P; and isotopes of sulfur, e.g. 35 S.

[0372] Certain isotopically labeled compounds of Formula I, for example, those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. In view of the ease of incorporation and the ready availability of detection means, the radioactive isotope tritium, i.e., 3 H, and carbon-14, i.e., 14 C is particularly useful for this purpose.

[0373] Using heavier isotopes, such as deuterium, 2 H substitution may offer certain therapeutic advantages due to its greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements.

[0374] Using positron emitting isotopes, e.g. 11 C. 18 F. 15 O and 13 N substitution can be used for positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0375] Isotopically-labeled compounds of Formula I can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

[0376] Pharmaceutically acceptable solvates according to the present invention include those in which the solvent of crystallization may be isotopically substituted (e.g., D 2 O、d 6 -acetone, d 6 -DMSO).

[0377] One way to implement the present invention is to administer a compound of formula I in the form of a prodrug. Thus, derivatives of certain compounds of formula I that have little or no pharmacological activity themselves can be converted into compounds of formula I having the desired activity when administered to or on the body, for example, by hydrolytic cleavage, particularly hydrolytic cleavage promoted by esterases or peptidases. Such derivatives are referred to as "prodrugs". Further information on the use of prodrugs can be found in: 'Pro-drugs as Novel Delivery Systems', Vol. 14, ACS Symposium Series (T. Higuchi and W. Stella) and 'Bioreversible Carriers in Drug Design', Pergamon Press, 1987 (Ed. EB Roche, American Pharmaceutical Association). Reference may also be made to Nature Reviews / Drug Discovery, 2008, 7, 355 and Current Opinion in Drug Discovery and Development, 2007, 10, 550.

[0378] Prodrugs according to the invention can, for example, be produced by replacing appropriate functional groups present in the compounds of formula I with certain moieties known to those skilled in the art as "pro-moieties", as described, for example, in 'Design of Prodrugs' by H. Bundgaard (Elsevier, 1985) and YM Choi-Sledeski and CG Wermuth, 'Designing Prodrugs and Bioprecursors', Practice of Medicinal Chemistry, (4th edition), Chapter 28, 657-696 (Elsevier, 2015).

[0379] Thus, the prodrugs according to the present invention are (a) ester or amide derivatives of carboxylic acids within the compounds of formula I; (b) ester, carbonate, carbamate, phosphate or ether derivatives of hydroxy groups within the compounds of formula I; (c) amide, imine, carbamate or amine derivatives of amino groups within the compounds of formula I; (d) oxime or imine derivatives of carbonyl groups within the compounds of formula I; or (e) methyl, primary alcohol or aldehyde groups that can be metabolically oxidized to carboxylic acids within the compounds of formula I.

[0380] Some specific examples of prodrugs according to the present invention include:

[0381] (i) Where the compound of formula I contains a carboxylic acid function (-COOH), an ester thereof, for example, wherein the hydrogen of the carboxylic acid function of the compound of formula I is replaced by C 1 -C 8 Alkyl (e.g., ethyl) or (C 1 -C 8 alkyl)C(=O)OCH 2 -(For example, t BuC(=O)OCH 2 -) substituted compounds;

[0382] (ii) In the case where the compound of formula I contains an alcohol function (-OH), an ester thereof, for example, wherein the hydrogen of the alcohol function of the compound of formula I is replaced by -CO(C 1 -C 8 Compounds in which an alcohol is esterified with an amino group;

[0383] (iii) In case the compound of formula I contains an alcohol function (-OH), its ether, for example, wherein the hydrogen of the alcohol function of the compound of formula I is replaced by (C 1 -C 8 alkyl)C(=O)OCH 2 -or-CH 2 OP(=O)(OH)2 Alternative compounds;

[0384] (iv) In the case where the compound of formula I contains an alcohol function (-OH), its phosphate ester, for example, wherein the hydrogen of the alcohol function of the compound of formula I is replaced by -P(=O)(OH) 2 or -P(=O)(ONa) 2 or -P(=O)(O - ) 2 Ca 2+ Alternative compounds;

[0385] (v) In the compounds of formula I containing a primary or secondary amino function (-NH 2 or -NHR, wherein R≠H), its amide, for example, wherein one or both (as the case may be) hydrogen atoms of the amino function of the compound of formula I are replaced by (C 1 -C 10 )alkanoyl, -COCH 2 NH 2 Compounds in which the amino group is substituted or derivatized with amino acids;

[0386] (vi) In the compound of formula I containing a primary or secondary amino function (-NH 2 or -NHR, wherein R≠H), in the case of an amine, for example, in which one or both (as the case may be) hydrogen atoms of the amino function of the compound of formula I are replaced by -CH 2 OP(=O)(OH) 2 Alternative compounds;

[0387] (vii) the carboxylic acid group in the compound of formula I is replaced by a methyl group, a -CH 2 In the case of OH group or aldehyde group replacement.

[0388] Certain compounds of Formula I can themselves act as prodrugs of other compounds of Formula I. Two compounds of Formula I can also be combined together in the form of prodrugs. In some cases, a prodrug of a compound of Formula I can be generated by internally linking two functional groups within a compound of Formula I (e.g., by forming a lactone).

[0389] Reference to a compound of formula I includes the compound itself and its prodrugs. The present invention includes such compounds of formula I as well as pharmaceutically acceptable salts of such compounds and pharmaceutically acceptable solvates of said compounds and salts.

[0390] Administration and dosing

[0391] In general, the compounds of the present invention are administered in an amount effective to treat the conditions described herein. The compounds of the present invention can be administered as the compounds themselves, or alternatively, in the form of pharmaceutically acceptable salts. For the purposes of administration and dosing, the compounds themselves or their pharmaceutically acceptable salts will simply be referred to as compounds of the present invention.

[0392] The compounds of the present invention can be administered by any appropriate route, in the form of a pharmaceutical composition adapted to such route, and in a dose effective for the intended treatment. The compounds of the present invention can be administered orally, rectally, vaginally, parenterally, or topically.

[0393] The compounds of the invention may be administered orally. Oral administration may involve swallowing, whereby the compound enters the gastrointestinal tract, or buccal or sublingual administration may be employed, whereby the compound enters the bloodstream directly from the mouth.

[0394] In another embodiment, the compounds of the invention may also be administered directly to the bloodstream, muscle, or internal organs. Suitable means for parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, and subcutaneous. Suitable devices for parenteral administration include needle (including microneedle) syringes, needle-free syringes, and infusion techniques.

[0395] In another embodiment, the compounds of the present invention may also be topically applied to the skin or mucous membranes, i.e., percutaneously or transdermally. In another embodiment, the compounds of the present invention may also be administered intranasally or by inhalation. In another embodiment, the compounds of the present invention may be administered rectally or vaginally. In another embodiment, the compounds of the present invention may also be administered directly to the eyes or ears.

[0396] The dosage regimen of the compounds of the present invention and / or compositions containing the compounds is based on various factors, including: the type, age, weight, sex and medical condition of the patient; the severity of the condition; the route of administration; and the activity of the specific compound used. Therefore, the dosage regimen can vary widely. In one embodiment, for the treatment of the specified condition discussed herein, the total daily dose of the compounds of the present invention is generally about 0.001 to about 100 mg / kg (i.e., the number of mg of the compounds of the present invention relative to each kg of body weight). In another embodiment, the total daily dose of the compounds of the present invention is about 0.01 to about 30 mg / kg, and in another embodiment, about 0.03 to about 10 mg / kg, and in yet another embodiment, about 0.1 to about 3 mg / kg. It is not uncommon to repeat multiple times (usually no more than four times) in one day to administer the compounds of the present invention. If necessary, multiple doses per day can usually be used to increase the total daily dose.

[0397] For oral administration, the composition can be provided in the form of tablets containing 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 30.0, 50.0, 75.0, 100, 125, 150, 175, 200, 250 and 500 mg of active ingredient to adjust the dose to the patient symptomatically. The drug typically contains about 0.01 mg to about 500 mg of active ingredient, or in another embodiment, about 1 mg to about 100 mg of active ingredient. When administered intravenously, the dose may be in the range of about 0.01 to about 10 mg / kg / minute during a constant rate infusion.

[0398] Suitable subjects according to the present invention include mammalian subjects. In one embodiment, humans are suitable subjects. Human subjects can be of either sex and any developmental stage.

[0399] Pharmaceutical composition

[0400] In another embodiment, the present invention comprises a pharmaceutical composition. Such a pharmaceutical composition comprises a compound of the present invention present together with a pharmaceutically acceptable carrier. Other pharmacologically active substances may also be present. "Pharmaceutically acceptable carrier" as used herein includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of the following: water, saline, phosphate-buffered saline, glucose, glycerol, ethanol, etc., and combinations thereof, and isotonic agents may be included in the composition, for example, sugars, sodium chloride, or polyols (e.g., mannitol or sorbitol). Pharmaceutically acceptable substances, for example, wetting agents or small amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives or buffers that improve the shelf life or effectiveness of the antibody or antibody portion.

[0401] The compositions of the present invention can be presented in various forms. These forms include, for example, liquid, semisolid and solid dosage forms, for example, liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories. The dosage form depends on the intended mode of administration and therapeutic application.

[0402] Typical compositions are in the form of injectable or infusible solutions, e.g., compositions similar to those used for passive immunization of humans containing general antibodies. One mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In another embodiment, the antibody is administered by intravenous infusion or injection. In yet another embodiment, the antibody is administered by intramuscular or subcutaneous injection.

[0403] Oral administration of solid dosage forms can be presented, for example, as discrete units, such as hard or soft capsules, pills, cachets, lozenges, or tablets, each containing a predetermined amount of at least one compound of the present invention. In another embodiment, oral administration can be in the form of a powder or granules. In another embodiment, the oral dosage form is sublingual, such as a lozenge. In such solid dosage forms, the compound of Formula I is generally combined with one or more adjuvants. Such capsules or tablets may contain a controlled release formulation. In the case of capsules, tablets, and pills, the dosage form may also contain a buffer or may be prepared using an enteric coating.

[0404] In another embodiment, oral administration may be in liquid dosage form. Liquid dosage forms for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art (e.g., water). Such compositions may also include adjuvants, for example, wetting agents, emulsifiers, suspending agents, flavoring agents (e.g., sweeteners), and / or spices.

[0405] In another embodiment, the present invention comprises parenteral dosage forms. "Parenteral administration" includes, for example, subcutaneous injection, intravenous injection, intraperitoneal injection, intramuscular injection, intrasternal injection, and infusion. Injectable preparations (i.e., sterile injectable aqueous or oily suspensions) can be prepared according to known techniques using appropriate dispersants, wetting agents, and / or suspending agents.

[0406] In another embodiment, the present invention comprises topical dosage forms. "Topical application" includes, for example, transdermal application (e.g., via a transdermal patch or iontophoresis device), intraocular application, or intranasal or inhaled application. Compositions for topical application also include, for example, topical gels, sprays, ointments, and creams. Topical preparations may include compounds that enhance the absorption or penetration of active ingredients through the skin or other affected areas. When the compounds of the present invention are applied via a transdermal device, a patch of a reservoir and a porous membrane type or a solid matrix variety is used to complete the application. Typical preparations for this purpose include: gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, skin patches, flat sheets, implants, sponges, fibers, bandages, and microemulsions. Liposomes may also be used. Typical carriers include: alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerol, polyethylene glycol, and propylene glycol. Permeation enhancers may be incorporated - see, for example, BC Finnin and TM Morgan, J. Pharm. Sci., vol. 88, pp. 955-958, 1999.

[0407] Preparations suitable for topical application to the eye include, for example, eye drops, wherein the compound of the present invention is dissolved or suspended in an appropriate carrier. Typical preparations suitable for eye or ear administration can be in the form of drops of isotonic, pH-adjusted, micronized suspensions or solutions in sterile saline. Other preparations suitable for eye and ear administration include: ointments, biodegradable (i.e., absorbable gel sponges, collagen) and non-biodegradable (i.e., silicone) implants, flat sheets, lenses and microgranular or vesicular systems (e.g., vesicles (niosomes) or liposomes). Polymers, for example, cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulose polymers (e.g., hydroxypropyl methylcellulose, hydroxyethyl cellulose or methylcellulose) or heteropolysaccharide polymers (e.g., gellan gum (gelan gum)), can be incorporated together with preservatives (e.g., benzalkonium chloride). This preparation can also be delivered by iontophoresis.

[0408] For intranasal administration or administration by inhalation, the compounds of the invention are conveniently delivered from a pump spray container (squeezed or pumped by the patient) in the form of a solution or suspension, or from a pressurized container or nebulizer using an appropriate propellant, in the form of an aerosol spray presentation. Formulations suitable for intranasal administration are typically in the form of a dry powder (alone, as a mixture (e.g., a dry blend with lactose), or as a mixed component granule (e.g., mixed with a phospholipid, such as lecithin)), administered by a dry powder inhaler; or as an aerosol spray, administered by a pressurized container, pump, spray, nebulizer (preferably a nebulizer using electrohydrodynamics to produce a fine mist), or nebulizer with or without an appropriate propellant (e.g., 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane). For intranasal use, the powder may contain a bioadhesive, for example, chitosan or cyclodextrin.

[0409] In another embodiment, the present invention comprises a rectal dosage form. Such a rectal dosage form can be in the form of, for example, a suppository. Cocoa butter is a traditional suppository base, but various substitutes can be used as appropriate.

[0410] Other carrier materials and modes of administration known in the pharmaceutical field may also be used. The pharmaceutical compositions of the present invention may be prepared by any well-known techniques of pharmacy, such as effective formulation and administration procedures. The foregoing considerations regarding effective formulation and administration procedures are well known in the art and have been described in standard textbooks. The formulation of drugs is discussed in, for example, Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, 1975; Liberman et al., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Kibbe et al., Eds., Handbook of Pharmaceutical Excipients (3rd Ed.), American Pharmaceutical Association, Washington, 1999.

[0411] Co-administration

[0412] The compounds of the present invention can be used alone or in combination with other therapeutic agents. The present invention provides any use, method or composition as defined herein, wherein any compound of formula I embodiment herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate of the compound or salt is used in combination with one or more other therapeutic agents discussed herein. This will include a pharmaceutical composition for treating a disease or condition adapted by an agonist of GLP-1R, comprising a compound of formula I, II, III, IV, or V as defined in any embodiment described herein, or a pharmaceutically acceptable salt thereof, and one or more other therapeutic agents discussed herein.

[0413] The administration of two or more compounds "in combination" means that all compounds are administered sufficiently closely in time so that each compound can produce a biological effect in the same time frame. The presence of one drug may change the biological effect of other compounds. Two or more compounds can be administered simultaneously, in parallel or sequentially. In addition, simultaneous administration can be performed by mixing the mixture before administration or in separate dosage forms at the same or different administration sites at the same time point.

[0414] "Concurrent administration," "co-administration," "simultaneous administration," and "administering simultaneously" mean that the compounds are administered in combination.

[0415] In another embodiment, the present invention provides a method of treatment comprising administering a compound of the present invention together with one or more additional drugs, wherein the one or more additional drugs can be selected from the agents discussed herein.

[0416] In one embodiment, the compounds of this invention are administered with an antidiabetic agent, including but not limited to: a biguanide (e.g., metformin); a sulfonylurea (e.g., tolbutamide, glibenclamide, gliclazide, chlorpropamide, tolazamide, acetohexamide, glyclopyramide, glimepiride, or glipizide); a thiazolidinedione (e.g., pioglitazone, rosiglitazone, or lobeglitazone); a glitazar (e.g., saroglitazar, aleglitazar, muraglitazar, or tesaglitazar); a meglitinide (e.g., glitazar, seroglitazar, aleglitazar, muraglitazar, or tesaglitazar); litinide (e.g., nateglinide, repaglinide); dipeptidyl peptidase 4 (DPP-4) inhibitors (e.g., sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, dutogliptin, or omarigliptin); glitazones (e.g., pioglitazone, rosiglitazone, balaglitazone, rivoglitazone, or lobeglitazone);Sodium-glucose linked transporter 2 (SGLT2) inhibitors (for example, empagliflozin, canagliflozin, dapagliflozin, ipragliflozin, Ipragliflozin, tofogliflozin, sergliflozin etabonate, remogliflozin etabonate) etabonate), or ergliptin); SGLTL1 inhibitors; GPR40 agonists (FFAR1 / FFA1 agonists, such as fasiglifam); glucose-dependent insulinotropic polypeptide (GIP) and its analogs; α-glucosidase inhibitors (such as voglibose, acarbose, or miglitol); insulin or insulin analogs; including pharmaceutically acceptable salts of specifically specified agents and pharmaceutically acceptable solvates of the agents and salts. ;

[0417] In another embodiment, the compounds of the invention are administered with an anti-obesity agent, including but not limited to: peptide YY or an analog thereof, a neuropeptide Y receptor type 2 (NPYR2) agonist, an NPYR1 or NPYR5 agonist, a cannabinoid receptor type 1 (CB1R) agonist, a lipase inhibitor (e.g., orlistat), human proislet peptide (HIP), a melanocortin receptor 4 agonist (e.g., setmelanotide), a melanin concentrating hormone receptor 1 antagonist, a farnesoid X receptor (FXR) agonist (e.g., obeticholic acid), a lipase inhibitor (e.g., orlistat), a ... (e.g., acid), zonisamide, phentermine (alone or in combination with topiramate), norepinephrine / dopamine reuptake inhibitors (e.g., buproprion), opioid receptor antagonists (e.g., naltrexone), combinations of norepinephrine / dopamine reuptake inhibitors and opioid receptor antagonists (e.g., buproprion in combination with naltrexone), GDG-15 analogs, Cholecystokinin agonists, amylin and its analogs (e.g., pramlintide), leptin and its analogs (e.g., metroleptin), serotonergic agents (e.g., lorcaserin), methionine aminopeptidase 2 (MetAP2) inhibitors (e.g., beloranib or ZGN-1061), phendimetrazine, diethylpropion, benzphetamine ( benzphetamine), SGLT2 inhibitors (e.g., empagliflozin, canagliflozin, dapagliflozin, ipragliflozin, ipragliflozin, togliflozin, sergliflozin etabonate, repagliflozin etabonate, or erpagliflozin), SGLTL1 inhibitors, dual SGLT2 / SGLT1 inhibitors, fibroblast growth factor receptor (FGFR) modulators, AMP-activated protein kinase (AMPK) activators, biotin, MAS receptor modulator, or a glucagon receptor agonist (alone or in combination with another GLP-1R agonist, for example, liraglutide, exenatide, dulaglutide, albiglutide, lixisenatide, or semaglutide); including pharmaceutically acceptable salts of the specifically specified agents and pharmaceutically acceptable solvates of the agents and salts.

[0418] In another embodiment, the compounds of the invention are administered in combination with one or more of the following: an agent for treating NASH (including, but not limited to, PF-05221304), a FXR agonist (e.g., obeticholic acid), a PPARα / δ agonist (e.g., elafibranor), a synthetic fatty acid-cholic acid conjugate (e.g., aramchol), a caspase inhibitor (e.g., emricasan), an anti-lysyl oxidase homolog 2 (LOXL2) monoclonal antibody (e.g., , simtuzumab), galectin 3 inhibitors (e.g., GR-MD-02), MAPK5 inhibitors (e.g., GS-4997), dual antagonists of chemokine receptor 2 (CCR2) and CCR5 (e.g., cenicriviroc), fibroblast growth factor 21 (FGF21) agonists (e.g., BMS-986036), leukotriene D4 (LTD4) receptor antagonists (e.g., tipelukast), niacin analogs (e.g., ARI 3037MO), ASBT inhibitors (e.g., volixibat), acetyl-CoA carboxylase (ACC) inhibitors (e.g., NDI010976 or PF-05221304), ketokinase (KHK) inhibitors, diacylglycerol acyltransferase 2 (DGAT2) inhibitors, CB1 receptor antagonists, anti-CB1R antibodies, or apoptosis signal regulating kinase 1 (ASK1) inhibitors, including pharmaceutically acceptable salts of the specifically indicated agents and pharmaceutically acceptable solvates of the agents and salts.

[0419] Certain specific compounds that can be used in combination with the compounds of the invention for treating the diseases or disorders described herein (e.g., NASH) include:

[0420] 4-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4'-piperidine]-1'-carbonyl)-6-methoxypyridin-2-yl)benzoic acid, which is an example of a selective ACC inhibitor and is prepared as a free acid in Example 9 of U.S. Pat. No. 8,859,577, which is the U.S. National Phase of International Application No. PCT / IB2011 / 054119, the disclosure of which is hereby incorporated by reference in its entirety for all purposes. Crystalline forms of 4-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4'-piperidine]-1'-carbonyl)-6-methoxypyridin-2-yl)benzoic acid, including an anhydrous mono-tris form (Form 1) and a trihydrate of the mono-tris salt (Form 2), described in International PCT Application No. PCT / IB2018 / 058966, the disclosure of which is hereby incorporated by reference in its entirety for all purposes;

[0421] (S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide, or a pharmaceutically acceptable salt thereof, and crystalline solid forms thereof (Form 1 and Form 2) are examples of DGAT2 inhibitors described in Example 1 of U.S. Pat. No. 10,071,992, the disclosure of which is hereby incorporated by reference in its entirety for all purposes;

[0422] [(1R,5S,6R)-3-{2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl}-3-azabicyclo[3.1.0]hexan-6-yl]acetic acid, or a pharmaceutically acceptable salt thereof (including its crystalline free acid form) is an example of a hexokinokinase inhibitor and is described in Example 4 of U.S. Pat. No. 9,809,579, the disclosure of which is hereby incorporated by reference herein in its entirety, the disclosure of which is hereby incorporated by reference herein in its entirety for all purposes; and

[0423] The FXR agonist Tropifexor, or a pharmaceutically acceptable salt thereof, is described in Example 1-1B of US Pat. No. 9,150,568, the disclosure of which is hereby incorporated by reference in its entirety for all purposes.

[0424] These agents and compounds of the invention may be combined with a pharmaceutically acceptable vehicle (eg, saline, Ringer's solution, dextrose solution, etc.) The specific dosage regimen, ie, dose, timing, and repetition, will depend on the particular individual and the individual's medical history.

[0425] Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and may include: buffers (e.g., phosphates, citrates, and other organic acids); salts (e.g., sodium chloride); antioxidants (including ascorbic acid and methionine); preservatives (e.g., octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride); chloride); benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens, e.g., methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins (e.g., serum albumin, gelatin, or Igs); hydrophilic polymers (e.g., polyvinyl pyrrolidone); amino acids (e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine); monosaccharides, disaccharides, and other carbohydrates (including glucose, mannose, or dextrins); chelating agents (e.g., EDTA); sugars (e.g., sucrose, mannitol, trehalose, or sorbitol); salt-forming counterions (e.g., sodium); metal complexes (e.g., zinc-protein complexes); and / or nonionic surfactants (e.g., TWEEN TM 、PLURONICS TM or polyethylene glycol (PEG).

[0426] Liposomes containing these agents and / or compounds of the invention are prepared by methods known in the art, for example, methods described in U.S. Patent Nos. 4,485,045 and 4,544,545. Liposomes with increased circulation time are disclosed in U.S. Patent No. 5,013,556. Particularly useful liposomes can be produced by reverse phase evaporation methods using lipid compositions comprising phosphatidylethanolamine (PEG-PE) derived from lecithin, cholesterol and PEG. Liposomes are extruded through filters with defined pore sizes to obtain liposomes with desired diameters.

[0427] These drugs and / or compounds of the invention may also be embedded in microcapsules prepared, for example, by coacervation techniques or interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacrylate) microcapsules, respectively; embedded in colloidal drug delivery systems (e.g., liposomes, albumin microparticles, microemulsions, nanoparticles and nanocapsules) or embedded in coarse emulsions. This technology is disclosed in Remington, The Science and Practice of Pharmacy, 20th Ed., Mack Publishing (2000).

[0428] Sustained release formulations can be used. Suitable examples of sustained release formulations include semipermeable matrices of solid hydrophobic polymers containing compounds of Formula I, II, III, IV, or V, in the form of molded articles, such as films or microcapsules. Examples of sustained release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactic acid (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and 7-ethyl-L-glutamine, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers (e.g., for LUPRON DEPOT TM (those in injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid.

[0429] Preparations for intravenous administration must be sterile. This can be easily accomplished by, for example, filtering through a sterile filtration membrane. The compounds of the invention are typically placed in a container with a sterile access port, for example, an intravenous solution bag or vial with a stopper that can be pierced by a hypodermic needle.

[0430] Suitable emulsions include commercially available fat emulsions, such as Intralipid TM , Liposyn TM Infonutrol TM 、Lipofundin TM Lipiphysan TM Preparation. The active ingredient may be soluble in a premixed emulsion composition or alternatively it may be soluble in an oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil or almond oil) and an emulsion formed by mixing a phospholipid (e.g., egg phospholipids, soybean phospholipids or soybean lecithin) and water. It is understood that other ingredients, such as glycerol or glucose, may be added to adjust the tension of the emulsion. Suitable emulsions typically contain up to 20% oil, e.g., 5 to 20%. Fat emulsions may contain fat droplets of 0.1 to 1.0 μm, particularly 0.1 to 0.5 μm, and have a pH in the range of 5.5 to 8.0.

[0431] The emulsion composition can be prepared by mixing the compound of the present invention with Intralipid TM Or those prepared by mixing their components (soybean oil, egg lecithin, glycerol and water).

[0432] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, and powders. Liquid or solid compositions may contain appropriate pharmaceutically acceptable excipients as listed above. In certain embodiments, the composition is administered by oral or nasal respiratory route to achieve local or systemic efficacy. Compositions in preferably sterile pharmaceutically acceptable solvents can be atomized using gases. Nebulized solutions can be inhaled directly by a nebulizing device or the nebulizing device can be connected to a mask, curtain or intermittent positive pressure respirator. Solutions, suspensions or powder compositions can be administered by a device that delivers the formulation in an appropriate manner, preferably orally or nasally.

[0433] Reagent test kit

[0434] Another aspect of the present invention provides a kit comprising a compound of formula I, II, III, IV, or V or a pharmaceutical composition comprising a compound of formula I, II, III, IV, or V of the present invention. In addition to a compound of formula I, II, or III of the present invention or a pharmaceutical composition thereof, the kit may also include a diagnostic or therapeutic agent. The kit may also include instructions for a diagnostic or therapeutic method. In certain embodiments, the kit comprises a compound of formula I, II, III, IV, or V or a pharmaceutical composition thereof and a diagnostic agent. In other embodiments, the kit comprises a compound of formula I, II, III, IV, or V or a pharmaceutical composition thereof.

[0435] In yet another embodiment, the invention comprises a kit suitable for carrying out the methods of treatment described herein. In one embodiment, the kit contains a first dosage form comprising one or more compounds of the invention in an amount sufficient to carry out the methods of the invention. In another embodiment, the kit comprises one or more compounds of the invention in an amount sufficient to carry out the methods of the invention and a container for the dosage.

[0436] preparation

[0437] The compounds of formula I, II, III, IV, or V can be prepared by the general and specific methods described below, using the common knowledge of those skilled in the art of synthetic organic chemistry. Such common knowledge can be found in standard reference books, for example, Comprehensive Organic Chemistry, Ed. Barton and Ollis, Elsevier; Comprehensive Organic Transformations: A Guide to Functional Group Preparations, Larock, John Wiley and Sons; and Compendium of Organic Synthetic Methods, Vol. I-XII (published by Wiley-Interscience). The starting materials used herein are commercially available or can be prepared by routine methods known in the art.

[0438] In the preparation of compounds of Formula I, II, III, IV, or V, it is worth noting that some of the preparation methods described herein may require protection of distal functional groups (e.g., primary amines, amines, carboxyl groups in Formula I precursors). The need for such protection will vary depending on the nature of the distal functional group and the conditions of the preparation method. One skilled in the art can easily determine the need for such protection. The use of such protection / deprotection methods is also a skill in the art. A general description of protecting groups and their uses is provided in: TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0439] For example, some compounds contain primary amine or carboxylic acid functional groups, which, if unprotected, may interfere with reactions at other sites of the molecule. Therefore, such functional groups may be protected by appropriate protecting groups, which may be removed in subsequent steps. Protecting groups suitable for amine and carboxylic acid protection include those commonly used in peptide synthesis (e.g., N-tert-butyloxycarbonyl (Boc), benzyloxycarbonyl (Cbz), and 9-fluorenylmethoxycarbonyl (Fmoc) for amines; and lower alkyl esters or benzyl esters for carboxylic acids), which are generally chemically non-reactive under the reaction conditions described and can generally be removed without chemically altering other functional groups in the compound of Formula I.

[0440] The schemes described below are intended to provide a general description of the methods used in the preparation of the compounds of the present invention. Certain 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 performed in a similar manner, regardless of whether the material is enantiomerically enriched or racemic. In addition, separation into the desired optically active materials can be performed at any desired point in the sequence using well-known methods (e.g., as described herein and in the chemical literature). For example, intermediates (e.g., S4, S7, S8, S24, S40, and S41) and final products (e.g., S25 and S42) can be separated using chiral chromatography. Alternatively, chiral salts can be utilized to separate enantiomerically enriched intermediates and final compounds.

[0441] In the following scenario, variables X, Y, and Z are 1 , Z 2 , Z 3 , R, R 1 , R 2 , R 3 , R 4 , m, p, and q are as defined herein for compounds of Formula I, II, III, IV, or V, unless otherwise indicated. For simplicity, the variable A is used to represent Ring A and its optional substituents R 1 . For the scheme provided below, X 1 , X 2 , X 3 , and X 4 Each may independently be a leaving group, for example, any alkyl or aryl sulfonate (eg, mesylate, tosylate, or triflate), or a halogen or any other group that can be replaced by an amine or used in a metal-mediated coupling reaction. 4 It can also be a protected carboxylic acid (ie, an ester). When the protecting group is identified as Pg 1 When the protecting group is identified as Pg, it can be an alkylamine protecting group (e.g., benzyl, benzhydryl, allyl, etc.); a carbamate protecting group (e.g., Boc, Cbz, etc.); or an amide protecting group (e.g., trifluoroacetamide). 2 When the protecting group is identified as Pg 3 When R is an alcohol protecting group (e.g., trimethylsilylethoxyethyl); or an acyl group (e.g., acetyl, benzoyl, etc.); or a trialkylsilyl group (e.g., trimethylsilyl, tert-butyldimethylsilyl, triisopropylsilyl, etc.). 2a Is H or -C 1-2 Alkyl, wherein the alkyl group may have 0 to 1 OH.4a It is C 1-2 Alkyl, C 0-2 Alkylene-C 3-6 Cycloalkyl, C 0-2 Alkylene-R 5 , or C 1-2 Alkylene-R 6 , wherein if valence permits, the alkyl, alkylene, or cycloalkyl group may be independently substituted by 0 to 3 F atoms and 0 to 1 independently selected C 0-1 Alkylene-OR O and -N(R N ) 2 substituted by a substituent.

[0442] Substituted piperidine S8 (wherein R 2 =H and Y=CH) can be prepared as discussed in Scheme 1. Aryl or heteroaryl bromide S1 can be treated with an alkyl lithium (e.g., butyl lithium or tert-butyl lithium) to give an aryl- or heteroaryl-lithium species, which can react with aldehyde S2 to give diol S3. Other aryl or heteroaryl organometallic reagents (e.g., but not limited to, Grignard reagents) can also be used to prepare S3. The reaction is typically carried out at a temperature of about -70°C. Then, NaIO 4 Oxidation of diol S3 to provide acetal S4 (R 2 =H). Compound S4 can then be reacted with a substituted boronic acid or boronic ester (S5) in the presence of a palladium catalyst and a ligand complex in the manner of the Suzuki reaction (Maluenda and Navarro, Molecules, 2015, 20, 7528-7557) to provide compounds of general formula S6. Reduction of the olefin to provide compounds of general structure S7 can be carried out under a hydrogen atmosphere (15-100 psi H 2 ), an alcoholic solvent (e.g., methanol or ethanol) or alternatively an aprotic organic solvent (e.g., ethyl acetate or THF), a suitable catalyst (e.g., palladium on carbon, Pd(OH) on carbon) 2 (Pearlman catalyst), PtO 2 (Adams catalyst), or tris(triphenylphosphine)rhodium(I) chloride (Wilkinson catalyst). Using an appropriate catalyst, a transfer hydrogenation reagent (e.g., ammonium formate or dihydrobenzene, or the like) may be employed. Alternatively, the reduction may be accomplished by alternative methods known to those skilled in the art using reagents (e.g., triethylsilane or other silanes) in the presence of an acid or metal catalyst, or using a metal reducing agent (e.g., magnesium or the like). Alternatively, the olefin may be functionalized by methods known to those skilled in the art to introduce R 3For example, olefins can be hydroborated to produce alcohols, which can be alkylated or further converted to nitriles, F or alkyl groups. 1 Removal of can be performed using a variety of methods described in the literature to provide amine S8.

[0443] Solution 1

[0444]

[0445] Scheme 2 provides an alternative preparation of compounds of general structure S4. The reaction of an appropriately substituted diol of general structure S9 with an aldehyde or ketone of general structure S10a in the presence of an acid (e.g., p-toluenesulfonic acid or pyridinium p-toluenesulfonate) in an aprotic organic solvent (e.g., toluene or benzene) can provide compounds of general structure S4. The reaction is typically heated at reflux using a Dean-Stark trap to remove water azeotropically. Diol S9 can also be reacted with a cyclic (dashed line present) or acyclic (dashed line absent) acetal or ketal of general structure S10b under acidic catalysis. The same is also applicable to the use of cyclic or acyclic thioacetals or thioketals of general structure S10c under the influence of mercuric salts, mild oxidants or alkylating agents to provide compound S4. Alternatively, the diol of general formula S9 can be reacted with an appropriately substituted alkyne S11 in an aprotic solvent (e.g., toluene) in the presence of triruthenium dodecacarbonyl at a temperature of about 100° C. to give a compound of general structure S4 (where R 2 =CH 2 R 2a ). In which R 2 Containing alcohol functional groups (e.g., CH 2 OH), the alcohol protecting group (Pg 3 ) (e.g., acetate) can be incorporated into compounds of general structure S10. The protecting group can then be removed in a subsequent step. Intermediate S4 can then be further modified using the methods described for Scheme 1 to provide amines of general structure S8.

[0446] Solution 2

[0447]

[0448] As provided in Scheme 3, conversion of S4 to compounds of general structure S7 (wherein Y = N) can be accomplished by, for example, Buchwald-Hartwig CN coupling between compounds of general structure S4 and appropriately substituted and protected piperazine S12 in the presence of a palladium or copper catalyst and a ligand complex. These reactions are typically carried out at 0 to 110° C. in the presence of a base (e.g., Cs 2 CO 3, LiHMDS or NaOtBu) (for example, but not limited to, 1,4-dioxane and PhCH 3 ) is carried out. 1 Removal of can be performed using a variety of methods described in the literature to provide amine S8 (wherein Y = N).

[0449] Solution 3

[0450]

[0451] Amine compound S8 prepared by the method described in Scheme 1-3 can be used to react with protected 2-bromoacetate in the presence of an appropriate base (e.g., K 2 CO 3 , Et 3 N, NaH or LiHMDS) in a polar aprotic solvent (such as but not limited to DMF, DMAc, DMSO or NMP) to give a compound of general structure S13 (X = N, L = CH 2 ). Standard ester hydrolysis can be performed to provide the acid S14. 2 If Pg is tert-butyl, standard acidic deprotection methods (e.g., TFA / DCM, HCl / 1,4-dioxane, HCl / EtOAc or other appropriate conditions) can be used to provide the acid S14. 2 If S14 is methyl or ethyl, standard basic deprotection methods (eg, aqueous NaOH in methanol or ethanol, or other appropriate conditions) can be used to provide acid S14.

[0452] Solution 4

[0453]

[0454] Compounds of general structure S15 (Scheme 5) can be precipitated in a base (e.g., sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate, potassium bicarbonate, cesium bicarbonate, sodium hydroxide, potassium hydroxide, cesium hydroxide, sodium acetate, potassium acetate, or cesium acetate) or an organic amine base (e.g., Et 3 N, DIPEA, DBU, etc.), in a polar aprotic solvent (such as but not limited to THF, DMF, DMAc, DMSO or NMP) or a protic solvent (such as water, MeOH, EtOH or iPrOH) or a mixture thereof, with an amine R 4 NH 2 Reaction to give compounds of general structure S16. It should be noted that if the examples provide R with a resolved enantiomeric center 4 , then other enantiomers or their racemic mixtures can be obtained by selecting appropriate raw materials. Preferred X 3Substituents include F, Cl, and Br, and preferred X 4 Groups include Cl, Br, and -CO 2 -Pg 2 The reduction of the nitro group can be carried out by using a metal catalyst (e.g., palladium on carbon or Raney nickel) in a protic solvent (e.g., methanol or ethanol) or an aprotic solvent (e.g., DMF, THF, or EtOAC) at 1-6 atm H 2 Alternatively, the nitro group can be hydrogenated using iron, zinc, SnCl 2 or other suitable metals in an acidic medium (e.g., 1N HCl, AcOH, or aqueous NH 4 Cl (in THF or methanol)) to provide compounds of general structure S17 (Scheme 5a). Compounds such as S18 can be acylated with acyl halides in standard manner or with carboxylates via standard amide coupling protocols to provide compounds S19. Reduction to compounds S20 can be carried out under standard conditions using a reducing agent (e.g., LAH or BH 3 -THF or BH 3 -DMS) (Scheme 5b).

[0455] Solution 5

[0456]

[0457] Diamine compounds S17 and S20 prepared by the methods described in Schemes 5a and 5b, collectively referred to as diamine S21 (Scheme 6), can be acylated using acids of general structure S14 under standard amide coupling protocols to give amides S22, which exist as a mixture of 100% S22a to 100% S22b. This mixture of amines S22 can be cyclized by various methods to give compounds of general structure S23. Amines S22 can be cyclized under microwave conditions (10-60 minutes at 120-180°C) with a dehydrating agent (e.g., ) or an alkanol (e.g., n-butanol) to provide compound S23. Alternatively, the mixture of compound S22 can be heated under acidic conditions (e.g., AcOH, 60-100° C.) or basic conditions (e.g., aqueous NaOH or KOH (in 1,4-dioxane), 60-100° C.) to provide S23. Compounds of general structure S23 (X 4 =Cl, Br or I) can be converted to an ester of structure S24 by palladium-catalyzed carbonylation using a suitable alcohol (e.g., MeOH or EtOH or other alkanol) under a 15-100 psi carbon monoxide atmosphere at a temperature of 20-100°C. Hydrolysis of ester S24 can be carried out as described in Scheme 4 to provide acid S25. For compounds S22 (wherein X 4=CO 2 -Pg 2 ), conversion to ester S24 is carried out under similar conditions as described above, except that a basic cyclization method is used, wherein compound S25 can be isolated directly from the reaction mixture. 4 It is CO 2 tBu), deprotection to the acid S25 can be carried out under the acidic conditions described in Scheme 4. Alternatively, for compound S24 (where Pg 2 It is C 1 -C 8 For example, alkyl, such as methyl, ethyl, hexyl or octyl, ester deprotection can be carried out using various enzymes well known to those skilled in the art, including esterases, proteases, peptidases, lipases, and glycosidases. The hydrolysis reaction can also be carried out by treating the ester with an aqueous solution of 1,5,7-triazabicyclo[4.4.0]dec-5-ene at room temperature.

[0458] Solution 6

[0459]

[0460] In addition, diamine S21 can be converted to 2-chloromethylbenzimidazole S26 by several methods (Scheme 7). Treatment with 2-chloroacetyl chloride or chloroacetic anhydride in an aprotic solvent (e.g., 1,4-dioxane) followed by heating at 40-100 °C for 2-18 hours can afford the desired benzimidazole S26 (where Z 1 , Z 2 and Z 3 is CH). In which Z 1 , Z 2 and Z 3 Not all CR zIn the case of , after treatment with 2-chloroacetyl chloride in an aprotic solvent (e.g., 1,4-dioxane) for 30 minutes to 4 hours, the solvent is changed to an acidic medium (e.g., AcOH or TFA), followed by heating at 40-100° C. for 2-18 hours to provide the desired compound S26. Diamine S21 can also be treated with chloroacetic anhydride at a temperature of 0 to 80° C. in an aprotic solvent (e.g., but not limited to, 1,4-dioxane, THF, or MeCN), followed by heating at 60-100° C. for 2 to 18 hours to obtain the desired compound S26. In addition, diamine S21 can be treated with 2-chloro-1,1,1-trimethoxyethane in an aprotic solvent (e.g., but not limited to, 1,4-dioxane, THF, or MeCN) or in a protic solvent (e.g., MeOH or EtOH) in the presence of an acid catalyst (e.g., pTSA) at 20-100° C. Alternatively, diamine S21 can be heated with 2-hydroxyacetic acid at 100-180° C. in an aprotic solvent (such as, but not limited to, mesitylene) to provide a hydroxymethyl intermediate. Conversion of the hydroxymethyl group to the chloromethyl compound S26 can be accomplished by standard methods, including reaction with SOCl in an aprotic solvent. 2 The reaction can be completed in a base (e.g., sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate, potassium bicarbonate, or cesium bicarbonate, NaH) or an organic amine base (e.g., Et 3 Compound S8 is reacted with a compound of general structure S26 in the presence of N, DIPEA, DBU, etc. in a polar aprotic solvent (such as, but not limited to, THF, MeCN, DMF, DMAc, DMSO or NMP) to obtain compound S23 (X 4 =Cl, Br, I) or compound S24 (X 4 =CO 2 -Pg 2 ), which was then used to obtain compound S25 via the method described in Scheme 6.

[0461] Solution 7

[0462]

[0463] Alternatively (Scheme 8), compounds of general structure S26 can be reacted in a base (e.g., sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate, potassium bicarbonate, or cesium bicarbonate, NaH) or an organic amine base (e.g., Et 3 N, DIPEA, DBU, etc.) in a polar aprotic solvent (such as, but not limited to, THF, MeCN, DMF, DMAc, DMSO or NMP) to provide compounds S27 (Scheme 8). 1Removal of can be performed using a variety of methods described in the literature to provide amine S28. General Structure S23 (X 4 =Cl, Br or I) or S24(X 4 =CO 2 -Pg 2 ) can be accomplished by a Buchwald-Hartwig CN coupling such as between compounds of general structure S4 and as described previously in Scheme 3. Compounds of general structure S23 or S24 can then be used to obtain compounds of structure S25 via the methods described in Scheme 6.

[0464] Solution 8

[0465]

[0466] Compounds of general structure S25 can also be prepared as discussed in Scheme 9. Diol S9 can be protected to provide S29. The trimethylsilylethoxymethyl group is a preferred protecting group. Protection of the diol as the corresponding acetal (e.g., formaldehyde acetal) is also preferred. Compound S29 can then be reacted with a substituted boronic acid or boronic ester (S5), and the olefin is then reduced using the method described in Scheme 1 to provide compounds of general structure S31 (wherein Y=CH). Alternatively, compound S29 can be coupled with piperazine of general structure S12 using the method described in Scheme 3 to provide S31 (wherein Y=N). Compounds of general structure S31 can be deprotected using the method described in Scheme 7 and then coupled with S26 to provide compounds of general structure 33. Alternatively, compounds of general structure S33 can be prepared from S32 by converting S32 to the corresponding N-acetic acid derivative as described in Schemes 4 and 6, and then condensing it with diamine S21. Deprotection of S33 using methods known to those skilled in the art can provide a diol of general structure S34, which can then be reacted with an alkyne of general structure S11 using the methods described in Scheme 2 to provide S23 or S24. Alternatively, S34 can be converted to S23 or S24 using an aldehyde, ketone, or derivative thereof as discussed in Scheme 2. Compounds of general structure S23 or S24 can then be used to obtain compounds of structure S25 via the methods described in Scheme 6.

[0467] Solution 9

[0468]

[0469] Compounds of general structure S24 and S33, where Y = N and XL = cyclopropyl, can be prepared as discussed in Scheme 10. Protected piperidone S35 can be homologated to unsaturated ester S36 using methods well known to those skilled in the art. For example, Horner-Wadsworth-Emmons olefination of S42 with a phosphate ester (e.g., ethyl (diethoxyphosphoryl)acetate) that has been deprotonated using a strong base (e.g., lithium tert-butoxide, sodium tert-butoxide, or potassium tert-butoxide) can provide S36. The reaction is typically carried out in an aprotic solvent such as THF or DME at a temperature around 0 to -50°C. Conversion of S36 to the cyclopropane derivative S37 can be accomplished by treatment with a sulfoxonium ylid derived from trimethylsulfoxonium iodide and a base (e.g., potassium tert-butoxide or sodium hydride). Deprotection of S37 and subsequent reaction of the resulting carboxylic acid S38 with S21 (where X 4 =CO 2 Pg 2 ) can provide compounds of general structure S39. Deprotection of S39 and coupling with S4 using the methods described in Scheme 3 can provide compounds of general structure S24 (wherein Y=N and XL is cyclopropyl). Compounds of general structure S24 can then be used to provide compounds of structure S25 via the methods described in Scheme 6. Alternatively, S40 can be reacted with S29 using the methods described in Scheme 3 to provide S33 (wherein Y=N and XL=cyclopropyl). Compounds of general structure S33 can then be used to provide compounds of structure S25 via the methods described in Scheme 6 and 9.

[0470] Solution 10

[0471]

[0472] Alternatively, compounds of general structure S25, wherein Y = N and XL is cyclopropyl, can be prepared as described in Scheme 11. Removal of Pg from S37 1 , to provide the piperidine derivative S43. Coupling of S43 with S4 in a manner similar to that described in Scheme 3 provides S13 (wherein Y=N and XL is cyclopropyl). Deprotection can then provide compounds of general structure S14, which can then be used to prepare S25 as described in Scheme 6.

[0473] Solution 11

[0474] BRIEF DESCRIPTION OF THE DRAWINGS

[0475] Figure 1This shows the observed powder X-ray diffraction pattern of the anhydrous (anhydrate) crystalline form (Form 1) of the tris salt of Compound Example 7.

[0476] Figure 2 This shows an observed powder X-ray diffraction pattern of the anhydrous (anhydrate) crystalline form (Form A) of the tris salt of Compound Example 10. DETAILED DESCRIPTION

[0477] The following illustrates the synthesis of non-limiting compounds of the present invention. Other compounds within the scope of the present invention can be prepared using the methods illustrated in these examples alone or in conjunction with techniques generally known in the art.

[0478] In general, experiments were performed under an inert atmosphere (nitrogen or argon), especially when reagents or intermediates sensitive to oxygen or moisture were used. Commercial solvents and reagents were usually used without further purification. Anhydrous solvents were used as necessary, typically Acros Organics Products from Sigma-Aldrich Sure / Seal TM or from EMD Chemicals In other cases, commercial solvents are filled molecular sieves until the following QC standards for water are reached: a) <100 ppm for dichloromethane, toluene, N,N-dimethylformamide, and tetrahydrofuran; b) <180 ppm for methanol, ethanol, 1,4-dioxane, and diisopropylamine. For very sensitive reactions, the solvents are further treated with sodium metal, calcium hydride, or molecular sieves and distilled just before use. The products are usually dried in vacuo before further reactions or submission to biological testing. Mass spectrometry data are reported by liquid chromatography-mass spectrometry (LCMS), atmospheric pressure chemical ionization (APCI), or gas chromatography-mass spectrometry (GCMS) instruments. The symbol ◆ indicates that a chlorine isotope pattern is observed in the mass spectrum.

[0479] Chiral separation is used to separate the enantiomers or diastereomers of certain intermediates during the preparation of the compounds of the present invention. When the chiral separation is completed, the separated enantiomers are designated as ENT-1 or ENT-2 (or DIAST-1 or DIAST-2) according to their elution order. In certain embodiments, the enantiomer designated as ENT-1 or ENT-2 can be used as a raw material to prepare other enantiomers or diastereomers. In this case, the enantiomers prepared are designated as ENT-X1 and ENT-X2 according to their raw materials; similarly, the diastereomers prepared are designated as DIAST-X1 and DIAST-X2 (or DIAST-) according to their raw materials. In the synthesis using multiple intermediates, the DIAST-Y and DIAST-Z nomenclatures are similarly used.

[0480] For compounds with two chiral centers, stereoisomers at each stereocenter are separated at different times. The designation of ENT-1 or ENT-2 (or DIAST-1 or DIAST-2) of an intermediate or embodiment refers to the elution order of the separation completed in the step. It can be recognized that when stereoisomers of a chiral center are separated in a compound with two or more centers, the enantiomers separated are diastereomers of each other. For example (but not limited to), Examples 15 and 16 have two chiral centers. When intermediate C36 is separated into ENT-1 (obtaining intermediate P17) and ENT-2 (obtaining intermediate P18), the chiral center of the cyclopropyl portion is separated. Then, P18 is used to prepare C70, which has a stereoisomer enriched in the chiral carbon of the cyclopropyl group and a mixture of stereoisomers on the dioxolane carbon. C70 was then separated into DIAST-Y1 on the dioxolane carbon to give intermediate C71, and DIAST-Y2 on the dioxolane carbon to give intermediate C72, wherein these intermediates were enriched in a single stereoisomer. C71 was then used to prepare Example 15, which was identified as 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, DIAST-X1, trifluoroacetate salt [starting from P18 via C71]. In these preparations, the chiral center was identified as "abs" near the center after the mixture was subjected to the separation procedure, understanding that the isolated enantiomers may not be enantiomerically pure. Typically, the enriched enantiomer at each chiral center is >90% of the isolate. Preferably, the enriched enantiomer at each center is >98% of the mixture.

[0481] In certain embodiments, the optical rotation of enantiomers is measured using a polarimeter. Based on its observed rotation data (or its specific rotation data), the enantiomer that rotates clockwise is designated as the (+)-enantiomer and the enantiomer that rotates counterclockwise is designated as the (-)-enantiomer. Racemic compounds are represented by undrawn or described stereochemistry, or by the appearance of (+ / -) next to the structure; in the latter case, the stereochemistry represented represents the relative (rather than absolute) configuration of the compound's substituents.

[0482] Reactions carried out through detectable intermediates are usually followed by LCMS and allowed to proceed to complete conversion before subsequent reagents are added. For syntheses with reference to procedures in other embodiments or methods, reaction conditions (reaction time and temperature) may vary. In general, reactions are followed by thin layer chromatography or mass spectrometry and, where appropriate, post-processing. Purification may vary from experiment to experiment: In general, solvents and solvent ratios for eluent / gradient are selected to provide appropriate R f s or retention time. All starting materials in these Preparations and Examples are commercially available or can be prepared by methods known in the art or as described herein.

[0483] Example

[0484] Preparation of P1

[0485] tert-Butyl 4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (P1)

[0486]

[0487] Step 1: Synthesis of 2-bromo-6-[(4-chloro-2-fluorophenyl)(hydroxy)methyl]phenol (C1)

[0488] This experiment was performed in two batches of the same scale. n-Butyl lithium (2.5 M solution in hexanes; 32.8 mL, 82.0 mmol) was slowly added to a -70°C solution of 1-bromo-4-chloro-2-fluorobenzene (17.2 g, 82.1 mmol) in diethyl ether (100 mL) while maintaining the temperature of the reaction mixture below -60°C. After stirring the reaction mixture at -70°C for 20 minutes, a solution of 3-bromo-2-hydroxybenzaldehyde (5.5 g, 27 mmol) in diethyl ether (100 mL) was slowly added while maintaining the reaction temperature below -60°C. After further stirring at -70°C for 1 hour, the reaction was quenched by the addition of aqueous ammonium chloride solution (50 mL) at -70°C, and the resulting mixture was diluted with water (100 mL). At this point, the two batches were combined and extracted with ethyl acetate (400 mL); the organic layer was washed with saturated aqueous sodium chloride solution (200 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (Gradient: 0% to 7% ethyl acetate in petroleum ether) provided C1 as a white solid. Combined yield: 15.7 g, 47.4 mmol, 88%. 1 H NMR (400MHz, chloroform-d) δ7.44 (dd, J=8.0, 1.5Hz, 1H), 7.37 (dd, J=8.1, 8.1Hz, 1H), 7.15 (br dd,J=8.5,2.1Hz,1H),7.12-7.05(m,2H),6.80(dd,J=7.8,7.8Hz,1H),6.78(s,1H),6.31(d,J=4.8Hz,1H),3.02(br d,J=4.9Hz,1H).

[0489] Step 2: Synthesis of 4-bromo-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxole (C2)

[0490] A solution of sodium periodate (25.4 g, 119 mmol) in water (105 mL) was added to a solution of C1 (15.7 g, 47.4 mmol) in methanol (450 mL), and the reaction mixture was stirred at 30 ° C for 16 hours, whereupon it was concentrated in vacuo. After diluting the residue with dichloromethane (500 mL), it was washed with water (500 mL). The dichloromethane solution was then dried over sodium sulfate, filtered, and concentrated in vacuo. Purification via silica gel chromatography (eluent: petroleum ether) provided C2 as a white solid. Yield: 10.0 g, 30.3 mmol, 64%. The following 1 H NMR data were obtained from experiments performed in the same manner but on a smaller scale. 1 H NMR (400 MHz, DMSO-d6 )δ7.67-7.61(m,2H),7.50(s,1H),7.43(br dd,J=8,2Hz,1H),7.09(dd,J=8.3,1.1Hz,1H),7.01(dd,J=7.9,1.1Hz,1H),6.86(dd,J=8.1,8.1Hz,1H).

[0491] Step 3: Synthesis of tert-butyl 4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]-3,6-dihydropyridine-1(2H)-carboxylate (C3)

[0492] A mixture containing C2 (8.00 g, 24.3 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (9.01 g, 29.1 mmol), sodium carbonate (5.15 g, 48.6 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride [Pd(dppf)Cl 2 ; 888mg, 1.21mmol] The reaction flask of the suspension in 1,4-dioxane (80mL) and water (32mL) was evacuated and charged with nitrogen. The evacuation cycle was repeated twice, and then, at 90°C, the reaction mixture was stirred for 16 hours. After the solvent was removed in vacuo, the residue was distributed between ethyl acetate (200mL) and water (200mL). The organic layer was washed with saturated aqueous sodium chloride solution (100mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Silica gel chromatography (gradient: 0% to 4.3% ethyl acetate (in petroleum ether)) provided the product, which was combined with a material from a similar reaction using C2 (2.00g, 6.07mmol) to obtain C3 as a light yellow jelly. Combined yield: 10.3g, 23.8mmol, 78%. 1 H NMR (400MHz, chloroform-d) δ7.53(dd,J=8.3,7.8Hz,1H),7.23-7.16(m,3H),6.88-6.83(m,2H),6.81-6.76(m,1H),6.34-6.28(br m,1H),4.10-4.05(m,2H),3.61(br dd,J=6,5Hz,2H),2.59-2.50(br m,2H),1.48(s,9H).

[0493] Step 4: Synthesis of tert-butyl 4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (P1)

[0494] A solution of C3 (10.3 g, 23.8 mmol) and tris(triphenylphosphine)rhodium(I) chloride (Wilkinson's catalyst; 1.54 g, 1.66 mmol) in methanol (100 mL) was stirred at 50 °C under hydrogen (45 psi) for 18 hours. The reaction mixture was then filtered through a pad of celite, and the filtrate was concentrated and subjected to silica gel chromatography (gradient: 0% to 9% ethyl acetate in petroleum ether) under reduced pressure. The resulting material was combined with material from a similar reaction performed using C3 (1.67 g, 3.87 mmol) to give P1 as a colorless gum. Combined yield: 10.3 g, 23.7 mmol, 86%. LCMS m / z 456.1 ◆ [M+Na + ]. 1 H NMR (400MHz, chloroform-d) δ7.52(dd,J=8.5,7.6Hz,1H),7.23-7.17(m,2H),7.16(s,1H),6.83(dd,J=7.8,7.8Hz,1H),6.78-6.69(m,2H),4.35-4.10(br m,2H),2.89-2.71(m,3H),1.89-1.77(m,2H),1.77-1.63(m,2H),1.47(s,9H).

[0495] Preparation of P2

[0496] tert-Butyl 4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (P2)

[0497]

[0498] Step 1: Synthesis of 4-bromo-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxole (C4)

[0499] 1-(4-Chloro-2-fluorophenyl)ethanone (316 g, 1.83 mol) and p-toluenesulfonic acid (6.02 g, 35.0 mmol) were added to a solution of 3-bromobenzene-1,2-diol (330 g, 1.75 mol) in toluene (1.5 L). The reaction apparatus was equipped with a Dean-Stark trap, and the reaction mixture was heated at 140° C. for 60 hours, whereupon the solution was concentrated in vacuo and purified using silica gel chromatography (eluent: petroleum ether); C4 was obtained as a mixture of yellow oil and solid. Yield: 158 g, 460 mmol, 26%. 1H NMR (400 MHz, chloroform-d): δ7.54 (dd, J=8.4, 8.4 Hz, 1H), 7.17-7.10 (m, 2H), 6.95 (dd, J=7.9, 1.4 Hz, 1H), 6.75 (dd, component of ABX pattern, J=7.8, 1.4 Hz, 1H), 6.70 (dd, component of ABX pattern, J=7.9, 7.9 Hz, 1H), 2.11 (d, J=1.1 Hz, 3H).

[0500] Step 2: Synthesis of tert-butyl 4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-3,6-dihydropyridine-1(2H)-carboxylate (C5)

[0501] Tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (62 g, 200 mmol) and sodium carbonate (100 g, 940 mmol) were added to a solution of C4 (58.0 g, 169 mmol) in 1,4-dioxane (600 mL). After adding [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) (6.0 g, 8.2 mmol), the reaction mixture was heated to 90 °C and stirred for 16 hours. Then, water (500 mL) was added, and the resulting mixture was extracted with ethyl acetate (2 x 500 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (2 x 500 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (Gradient: 0% to 9% ethyl acetate in petroleum ether) provided C5 as a yellow oil. Yield: 56.0 g, 126 mmol, 75%. 1 H NMR (400 MHz, chloroform-d) δ 7.50 (dd, J = 8.2, 8.2 Hz, 1H), 7.17-7.09 (m, 2H), 6.83-6.77 (m, 2H), 6.74 (dd, component of ABX pattern, J = 5.4, 3.6 Hz, 1H), 6.39-6.33 (br m, 1H), 4.14-4.08 (m, 2H), 3.70-3.56 (m, 2H), 2.66-2.45 (m, 2H), 2.07 (d, J = 1.1 Hz, 3H), 1.50 (s, 9H).

[0502] Step 3: Synthesis of tert-butyl 4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (P2)

[0503] Tris(triphenylphosphine)rhodium(I) chloride (Wilkinson's catalyst; 8.10 g, 8.75 mmol) was added to a solution of C5 (56.0 g, 126 mmol) in methanol (200 mL), and the reaction mixture was heated to 50° C. for 18 hours under hydrogen (45 psi). It was then cooled to 25° C. and filtered through celite. The filtrate was concentrated in vacuo and purified twice using silica gel chromatography (first column - gradient: 0% to 9% ethyl acetate (in petroleum ether); second column - gradient: 0% to 2% ethyl acetate (in petroleum ether)) to give P2 as a yellow solid. Yield: 37.0 g, 82.6 mmol, 66%. LCMS m / z 392.1 ◆ [(M-2-methylprop-1-ene) + H] + . 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.51 (dd, J = 8.3, 8.0 Hz, 1H), 7.17-7.09 (m, 2H), 6.77 (dd, component of ABC pattern, J = 7.8, 7.8 Hz, 1H), 6.70 (dd, component of ABC pattern, J = 7.7, 1.3 Hz, 1H), 6.66 (dd, component of ABC pattern, J = 7.8, 1.3 Hz, 1H), 4.37-4.13 (br m, 2H), 2.92-2.73 (m, 3H), 2.05 (d, J = 1.1 Hz, 3H), 1.90-1.63 (m, 4H), 1.49 (s, 9H).

[0504] Preparation of P3

[0505] 4-[(2S)-2-(4-Chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine, p-toluenesulfonate (P3)

[0506]

[0507] Step 1: Separation of tert-butyl 4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (C6) and tert-butyl 4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (C7)

[0508] P2 (75.2 g, 168 mmol) was separated into its constituent enantiomers via SFC (supercritical fluid chromatography) [column: Chiral Technologies Chiralpak AD-H, 5 μm; mobile phase: 4:1 carbon dioxide / (2-propanol containing 0.2% 1-aminopropan-2-ol)]. The first eluting compound was designated as C6 and the second eluting enantiomer as C7. The indicated absolute configurations were assigned based on single crystal X-ray structure determinations performed on C8 (which is derived from C6, see below).

[0509] C6 - Yield: 38.0 g, 84.8 mmol, 50%. Retention time 3.64 min [Column: Chiral Technologies Chiralpak AD-H, 4.6 x 250 mm, 5 μm; Mobile phase A: carbon dioxide; Mobile phase B: 2-propanol containing 0.2% 1-aminopropan-2-ol; Gradient: 5% B in 1.00 min, then 5% to 60% B in 8.00 min; Flow rate: 3.0 mL / min; Back pressure: 120 bar].

[0510] C7 - Yield: 36.8 g, 82.2 mmol, 49%. Retention time 4.19 min (Analytical SFC conditions were the same as those used for C6).

[0511] Step 2: Synthesis of 4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine, p-toluenesulfonate (P3)

[0512] A solution of C7 (1.62 g, 3.62 mmol) in ethyl acetate (36 mL) was treated with p-toluenesulfonic acid monohydrate (791 mg, 4.16 mmol) and heated at 45 °C. After 23 hours, the reaction mixture was cooled to room temperature and the solid was collected by filtration. It was rinsed with a mixture of ethyl acetate and heptane (1:1, 2 x 15 mL) to give P3 as a white solid. Yield: 1.37 g, 2.63 mmol, 73%. LCMS m / z 348.1◆[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ8.53(v brs,1H),8.29(v br s,1H),7.65-7.55(m,2H),7.47(d,J=8.1Hz,2H),7.35(dd,J=8.4,2.0Hz,1H),7.11(d,J=7.8Hz,2H),6.88-6.81 (m,2H),6.75-6.68(m,1H),3.42-3.33(m,2H),3.11-2.93(m,3H),2.29(s,3H),2.03(s,3H),1.98-1.82(m,4H).

[0513] C6 was converted to 4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine, mesylate (C8) for determination of absolute stereochemistry

[0514]

[0515] p-Toluenesulfonic acid (377 mg, 2.19 mmol) was added to a solution of C6 (490 mg, 1.09 mmol) in ethyl acetate (5.5 mL), and the reaction mixture was stirred at room temperature overnight. After dilution with additional ethyl acetate, the reaction mixture was washed successively with aqueous sodium bicarbonate solution, water, and saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Yield: 375 mg, 1.08 mmol, 99%. 1 H NMR (400 MHz, methanol-d 4 )δ7.59(dd,J=8.3.8.3Hz,1H),7.27(dd,J=10.9,2.0Hz,1H),7.20(br dd,J=8.4,2.1Hz,1H),6.81-6.75(m,1H),6.74-6.67(m,2H),3.18-3.09(m,2H), 2.88-2.77(m,1H),2.77-2.67(m,2H),2.02(d,J=0.7Hz,3H),1.85-1.73(m,4H).

[0516] A 0.1 M solution of this free base (4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine) in ethyl acetate was prepared and subjected to salt screening. Only the formation of the mesylate salt is described here. A mixture of methanesulfonic acid (25 μL, 39 μmol) and substrate solution (0.1 M; 0.25 mL, 25 μmol) was stirred overnight. Sufficient methanol was then added to dissolve the solids present, and ethyl acetate (3 mL) was added. The resulting solution was slowly evaporated without stirring to give C8 crystals, one of which was used for the single crystal X-ray structure determination described below.

[0517] Single crystal X-ray structure determination of C8

[0518] Single crystal X-ray analysis

[0519] Data collection was performed at room temperature on a Bruker D8 Quest diffractometer. Data collection consisted of ω and φ scans.

[0520] Through intrinsic phasing, SHELX software was used to analyze the orthorhombic space group P2 1 2 1 2 1 The structure was solved in . Then, the structure was refined by full-matrix least squares. All non-hydrogen atoms were found and refined using anisotropic displacement parameters.

[0521] The formation of the mesylate salt was confirmed via N1_H1X_O4 proton transfer.

[0522] Hydrogen atoms located on nitrogen and oxygen were found from Fourier difference maps and refined with restrained distances. The remaining hydrogen atoms were placed at calculated positions and carried on their supporting atoms. The final refinement included isotropic displacement parameters for all hydrogen atoms.

[0523] Absolute structure analysis using the likelihood method (Hooft, 2008) was performed using PLATON (Spek). The results indicated that the absolute structure had been correctly assigned; the method calculated the probability that the structure was correct to be 100%. Hootf parameters were reported as 0.02, esd as 0.0012, and Parson parameters were reported as 0.07, esd as 0.009. The absolute configuration of C7 was confirmed to be (R).

[0524] The asymmetric unit is composed of one molecule of C8 protonated free base and one molecule of deprotonated methanesulfonic acid. The final R index is 4.6%. The final difference Fourier analysis shows no missing or misplaced electron density.

[0525] Relevant crystal, data collection, and refinement information are summarized in Table A. Atomic coordinates, bond lengths, bond angles, and displacement parameters are listed in Table BD.

[0526] Software and References

[0527] SHELXTL, Version 5.1, Bruker AXS, 1997.

[0528] PLATON,ALSpek,J.Appl.Cryst.2003,36,7-13.

[0529] MERCURY, CF Macrae, PR Edington, P. McCabe, E. Pidcock, GP Shields, R. Taylor, M. Towler and van de Streek, J. Appl. Cryst. 2006, 39, 453-457.

[0530] OLEX2, OV Dolomanov, LJ Bourhis, RJ Gildea, JAK Howard and Puschmann, J. Appl. Cryst. 2009, 42, 339-341.

[0531] RWWHooft, LHStraver and L.Spek, J.Appl.Cryst.2008,41,96-103.

[0532] HDFlack, Acta Cryst. 1983, A39, 867-881.

[0533] Table A: Crystal data and structure refinement of C8

[0534]

[0536] Table B: Atomic coordinates of C8 (x 10 4 ) and the equivalent isotropic displacement parameter

[0537] U(eq) is defined as the orthogonal U ij One third of the tensor's trace.

[0538]

[0539] Table C: Bond lengths of C8 and angle [°].

[0540]

[0541]

[0542]

[0543] Symmetry transformations used to produce equivalent atoms.

[0544] Table D: Anisotropic displacement parameters of C8 The anisotropic displacement factor exponent takes the following form: -2π 2 [h 2 a* 2 U 11 +...+2h ka*b*U 12 ].

[0545]

[0546] P3, Preparation of di-p-toluoyl-L-tartrate

[0547] 4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine, di-p-toluoyl-L-tartrate (P3, di-p-toluoyl-L-tartrate)

[0548]

[0549] A solution of C13, free base (519 mg, 1.49 mmol) and di-p-toluoyl-L-tartaric acid (278 mg, 0.719 mmol) in acetonitrile (7.5 mL) was stirred at 50 °C for 1.5 hours. The mixture was cooled to room temperature at 0.2 °C / min. After 15 hours at room temperature, the mixture was heated to 65 °C and charged with acetonitrile (15 mL). The mixture was cooled to room temperature at 0.2 °C / min. After 15 hours at room temperature, the mixture was heated to 54 °C. After 3 hours, the solid was collected by filtration and dried in a vacuum oven at 35 °C under nitrogen to provide P3, di-p-toluoyl-L-tartaric acid salt (217 mg, 0.296 mmol, 20%, 82% ee) as a white solid.

[0550] A solution of P3, di-p-toluoyl-L-tartrate (217 mg, 0.296 mmol, 20%, 82% ee) in acetonitrile (8.0 mL) was cooled to room temperature at 0.2°C / min at 50°C. After 15 hours, the solid was collected by filtration and dried in a vacuum oven under nitrogen at 35°C to give P3, di-p-toluoyl-L-tartrate (190 mg, 0.259 mmol, 88%, 88% ee) as a white solid. LCMS m / z 348.1◆[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ8.9-8.5 (br s, 2H), 7.79 (d, J=8.1 Hz, 4H), 7.64-7.54 (m, 2H), 7.34 (dd, J=8.4, 2.1 Hz, 1H), 7.26 (d, J=8.0 Hz, 4H), 6.87-6.78 (m, 2H), 6.69 (dd, J=6.7, 2.5 Hz, 1H), 5.58 (s, 2H), 3.37-3.28 (m, 2H, assumed; partially obscured by water peak), 3.05-2.89 (m, 3H), 2.33 (s, 6H), 2.02 (s, 3H), 1.92-1.80 (m, 4H). Retention times: Peak 1 (4.97 minutes, minor) and Peak 2 (5.31 minutes, major) {Column: Chiralpak IC-U 3.0 x 50 mm, 1.6 μm; Mobile phase A: carbon dioxide; Mobile phase B: 0.1% isopropylamine in methanol; Gradient: 10% B, 5.00 minutes; then 45% B, 0.6 minutes; Flow rate: 1.7 mL / min; Back pressure: 130 bar}.

[0551] Preparation of P4

[0552] tert-Butyl 4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (P4)

[0553] A suspension of P2 (2.00 g, 4.46 mmol), zinc cyanide (734 mg, 6.25 mol), zinc (70.1 mg, 1.07 mmol), 1,1'-bis(diphenylphosphino)ferrocene (dppf; 198 mg, 0.357 mmol) and tris(dibenzylideneacetone)dipalladium(0) (164 mg, 0.179 mmol) in N,N-dimethylacetamide (20 mL) was stirred at 120°C for 16 hours, whereupon it was filtered. The filtrate was mixed with water (50 mL) and extracted with ethyl acetate (3 x 50 mL); the combined organic layers were then washed successively with water (30 mL) and saturated aqueous sodium chloride solution (20 mL) and concentrated in vacuo. Silica gel chromatography (gradient: 0% to 30% ethyl acetate in petroleum ether) gave a solid, which was treated with acetonitrile (15 mL) and water (15 mL) and subjected to freeze drying. This gave P4 as a light yellow solid. Yield: 1.17 g, 2.67 mmol, 60%. LCMS m / z 461.3 [M+Na + ]. 1 H NMR (400 MHz, CHLOROFORM-d) 7.71 (dd, J = 7.7, 7.6 Hz, 1H), 7.45 (dd, J = 8.0, 1.6 Hz, 1H), 7.42 (dd, J = 10.0, 1.5 Hz, 1H), 6.79 (dd, component of ABC pattern, J = 7.7, 7.6 Hz, 1H), 6.72 (dd, component of ABC pattern, J = 7.8, 1.3 Hz, 1H), 6.68 (dd, component of ABC pattern, J = 7.8, 1.3 Hz, 1H), 4.37-4.14 (br m, 2H), 2.91-2.73 (m, 3H), 2.07 (d, J = 1.1 Hz, 3H), 1.89-1.62 (m, 4H), 1.49 (s, 9H).

[0554] Preparation of P5 and P6

[0555] 4-Bromo-2-phenyl-1,3-benzodioxole, ENT-1(P5) and 4-Bromo-2-phenyl-1,3-benzodioxole, ENT-2(P6)

[0556]

[0557] Step 1: Synthesis of 2-bromo-6-[hydroxy(phenyl)methyl]phenol (C9)

[0558] Phenyllithium (1.9 M solution in 1-butoxybutane; 78.5 mL, 149 mmol) was slowly added to a -70°C solution of 3-bromo-2-hydroxybenzaldehyde (10.0 g, 49.7 mmol) in tetrahydrofuran (70 mL) at a rate that maintained the reaction temperature below -60°C. The resulting suspension was stirred for 1 hour at -70°C and then allowed to warm to room temperature overnight, whereupon it was poured into an aqueous ammonium chloride solution (30 mL) at 0°C. The mixture was extracted with ethyl acetate (3x30 mL), and the combined organic layers were washed with saturated aqueous sodium chloride solution (30 mL), dried over sodium sulfate, filtered and concentrated in vacuo. Silica gel chromatography (gradient: 0% to 5% ethyl acetate in petroleum ether) provided C9 as a yellow solid. Yield: 6.11 g, 21.9 mmol, 44%. 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.45-7.28 (m, 6H), 7.22-7.18 (m, 1H), 7.06 (br d, J=7.7 Hz, 1H), 6.77 (dd, J=7.9, 7.8 Hz, 1H), 6.06 (br s, 1H), 2.89 (br s, 1H).

[0559] Step 2: Synthesis of 4-bromo-2-phenyl-1,3-benzodioxole (C10)

[0560] A solution of sodium periodate (11.7 g, 54.7 mmol) in water (175 mL) was added to a solution of C9 (6.11 g, 21.9 mmol) in methanol (370 mL). The reaction mixture was stirred at 30 ° C for 40 hours, whereupon most of the methanol was removed by vacuum concentration. The resulting mixture was extracted with dichloromethane (5 x 100 mL), and the combined organic layers were washed successively with aqueous sodium sulfite solution (100 mL) and saturated aqueous sodium chloride solution (100 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Silica gel chromatography (eluent: petroleum ether) provided C10 as a colorless oil. Yield: 4.50 g, 16.2 mmol, 74%. LCMS m / z 278.5 (bromine isotope pattern observed) [M+H] + . 1 H NMR (400 MHz, chloroform-d) δ 7.62-7.57 (m, 2H), 7.49-7.43 (m, 3H), 7.04 (s, 1H), 7.00 (dd, J = 8.0, 1.4 Hz, 1H), 6.79 (dd, component of ABX pattern, J = 7.8, 1.4 Hz, 1H), 6.75 (dd, component of ABX pattern, J = 7.9, 7.8 Hz, 1H).

[0561] Step 3: Separation of 4-bromo-2-phenyl-1,3-benzodioxole, ENT-1 (P5) and 4-bromo-2-phenyl-1,3-benzodioxole, ENT-2 (P6)

[0562] Using SFC [column: Chiral Technologies ChiralCel OD, 10 μm; mobile phase: 3:1 carbon dioxide / (methanol containing 0.1% ammonium hydroxide)], enantiomers including C10 (5.00 g, 18.0 mmol) were separated. The first eluting enantiomer was designated as ENT-1 (P5), and the second eluting enantiomer was designated as ENT-2 (P6); both were obtained as yellow oils.

[0563] P5 yield: 2.20 g, 7.94 mmol, 44%. LCMS m / z 277.0 (bromine isotope pattern observed) [M+H] + . 1 HNMR (400 MHz, chloroform-d) δ 7.63-7.55 (m, 2H), 7.51-7.42 (m, 3H), 7.04 (s, 1H), 7.00 (dd, J = 8.0, 1.3 Hz, 1H), 6.80 (dd, component of ABX mode, J = 7.8, 1.4 Hz, 1H), 6.75 (dd, component of ABX mode, J = 7.9, 7.8 Hz, 1H). Retention time 3.28 minutes (column: Chiral Technologies ChiralCel OD-H, 4.6x150mm, 5μm; mobile phase A: carbon dioxide; mobile phase B: methanol containing 0.05% diethylamine; gradient: 5% to 40% B, over 5.5 minutes; flow rate: 2.5 mL / min).

[0564] P6 yield: 2.00 g, 7.22 mmol, 40%. LCMS m / z 276.9 (bromine isotope pattern observed) [M+H] + . 1 HNMR (400 MHz, chloroform-d) δ 7.63-7.55 (m, 2H), 7.50-7.42 (m, 3H), 7.04 (s, 1H), 7.00 (dd, J = 8.0, 1.4 Hz, 1H), 6.80 (dd, component of ABX pattern, J = 7.8, 1.4 Hz, 1H), 6.75 (dd, component of ABX pattern, J = 7.9, 7.9 Hz, 1H). Retention time 3.73 minutes (analytical conditions are the same as those used for P5).

[0565] Preparation of P7

[0566] tert-Butyl 4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (P7)

[0567]

[0568] Step 1: Synthesis of 2-(4-bromo-2-methyl-1,3-benzodioxol-2-yl)-5-chloropyridine (C11)

[0569] A mixture of 5-chloro-2-ethynylpyridine (1.80 g, 13.1 mmol), 3-bromobenzene-1,2-diol (2.47 g, 13.1 mmol) and triruthenium dodecacarbonyl (167 mg, 0.261 mmol) in toluene (25 mL) was degassed for 1 minute and then heated at 100 °C for 16 hours. The reaction mixture was diluted with ethyl acetate (30 mL) and filtered through a pad of celite; the filtrate was concentrated under vacuum and purified using silica gel chromatography (gradient: 0% to 1% ethyl acetate in petroleum ether) to provide C11 as a yellow oil. Yield: 1.73 g, 5.30 mmol, 40%. LCMS m / z 325.6 (bromine-chlorine isotope pattern observed) [M+H] + . 1 H NMR (400 MHz, chloroform-d) δ8.63 (dd, J=2.4, 0.7 Hz, 1H), 7.71 (dd, component of ABX pattern, J=8.4, 2.4 Hz, 1H), 7.60 (dd, component of ABX pattern, J=8.4, 0.7 Hz, 1H), 6.97 (dd, J=8.0, 1.4 Hz, 1H), 6.76 (dd, component of ABX pattern, J=7.8, 1.4 Hz, 1H), 6.72 (dd, component of ABX pattern, J=8.0, 7.8 Hz, 1H), 2.10 (s, 3H).

[0570] Step 2: Synthesis of tert-butyl 4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]-3,6-dihydropyridine-1(2H)-carboxylate (C12)

[0571] [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II)dichloride (388 mg, 0.530 mmol) was added to a suspension of C11 (1.73 g, 5.30 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.64 g, 5.30 mmol), and cesium carbonate (5.18 g, 15.9 mmol) in 1,4-dioxane (35 mL) and water (6 mL). The reaction mixture was stirred at 90 °C for 4 hours, whereupon it was diluted with ethyl acetate (30 mL) and water (5 mL). The organic layer was concentrated in vacuo and the residue was subjected to silica gel chromatography (Gradient: 0% to 5% ethyl acetate in petroleum ether) to give C12 as a yellow gum. Yield: 1.85 g, 4.31 mmol, 81%. LCMS m / z 451.0◆[M+Na + ]. 1 H NMR (400 MHz, chloroform-d) δ 8.62 (dd, J = 2.5, 0.8 Hz, 1H), 7.69 (dd, component of ABX pattern, J = 8.4, 2.4 Hz, 1H), 7.57 (dd, component of ABX pattern, J = 8.4, 0.8 Hz, 1H), 6.84-6.79 (m, 2H), 6.78-6.73 (m, 1H), 6.39-6.33 (br m, 1H), 4.13-4.07 (m, 2H), 3.68-3.58 (m, 2H), 2.60-2.51 (br m, 2H), 2.07 (s, 3H), 1.49 (s, 9H).

[0572] Step 3: Synthesis of tert-butyl 4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (P7)

[0573] A solution of C12 (2.61 g, 6.08 mmol) and tris(triphenylphosphine)rhodium(I) chloride (Wilkinson's catalyst; 563 mg, 0.608 mmol) in methanol (100 mL) was degassed in vacuo and then purged with hydrogen; this evacuation-purge cycle was performed a total of three times. The reaction mixture was then stirred under hydrogen (50 psi) at 60 °C for 16 hours, whereupon it was filtered. The filtrate was concentrated in vacuo and the residue was purified using silica gel chromatography (gradient: 0% to 10% ethyl acetate in petroleum ether); the resulting material was combined with material from a similar hydrogenation performed on C12 (110 mg, 0.256 mmol) to provide P7 as a pale yellow gum. Combined yield: 2.05 g, 4.76 mmol, 75%. LCMS m / z 431.3 ◆ [M+H] + . 1 H NMR (400 MHz, chloroform-d) δ8.62 (d, J=2.3 Hz, 1H), 7.69 (dd, component of ABX pattern, J=8.4, 2.4 Hz, 1H), 7.57 (d, half of AB quartet, J=8.4 Hz, 1H), 6.79 (dd, component of ABC pattern, J=7.8, 7.7 Hz, 1H), 6.72 (dd, component of ABC pattern, J=7.8, 1.3 Hz, 1H), 6.68 (br d, component of ABC pattern, J=7.9 Hz, 1H), 4.32-4.12 (br m, 2H), 2.91-2.73 (m, 3H), 2.05 (s, 3H), 1.90-1.62 (m, 4H), 1.48 (s, 9H).

[0574] Preparation of P8 and P9

[0575] 4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylic acid tert-butyl ester, ENT-1(P8) and 4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylic acid tert-butyl ester, ENT-2(P9)

[0576]

[0577] Separation of P7 (500 mg, 1.16 mmol) into its constituent enantiomers was performed using SFC {column: Phenomenex Lux Amylose-1, 5 μm; mobile phase: 9:1 carbon dioxide / [2-propanol containing 0.2% (7 M ammonia in methanol)]}. The first eluting enantiomer was designated as ENT-1 (P8), and the second eluting enantiomer was designated as ENT-2 (P9).

[0578] P8 yield: 228 mg, 0.529 mmol, 46%. Retention time 4.00 min {column: Phenomenex LuxAmylose-1, 4.6 x 250 mm, 5 μm; mobile phase A: carbon dioxide; mobile phase B: [2-propanol containing 0.2% (7 M ammonia in methanol)]; gradient: 5% B, 1.00 min, then 5% to 60% B, over 8.00 min; flow rate: 3.0 mL / min; back pressure: 120 bar}.

[0579] P9 yield: 229 mg, 0.531 mmol, 46%. Retention time 4.50 min (analytical conditions were the same as those used for P8).

[0580] Preparation of P10

[0581] {4-[2-(4-Chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}acetic acid (P10)

[0582]

[0583] Step 1: Synthesis of 4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine, p-toluenesulfonate (C13)

[0584] A solution of P2 (5.0 g, 11 mmol) and p-toluenesulfonic acid (4.81 g, 27.9 mmol) in ethyl acetate (100 mL) was stirred at 60 °C for 2 hours, whereupon it was concentrated in vacuo to afford C13 as a yellow gum. This material was used directly in the next step. LCMS m / z 347.9◆[M+H] + .

[0585] Step 2: Synthesis of ethyl {4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}acetate (C14)

[0586] Potassium carbonate (7.71 g, 55.8 mmol) and ethyl bromoacetate (1.86 g, 11.2 mmol) were added to a solution of C13 (from the previous step; ≤ 11 mmol) in acetonitrile (150 mL), and the reaction mixture was stirred at 55 °C for 16 hours. It was then filtered, and the filtrate was concentrated in vacuo and purified using silica gel chromatography (Gradient: 0% to 30% ethyl acetate in petroleum ether) to give C14 as a yellow gum. 1 H NMR analysis showed that this material was not completely pure. Yield: 3.57 g, 8.23 ​​mmol, 75% (over two steps). 1 H NMR (400 MHz, chloroform-d), C14 peak only: δ 7.52 (dd, J = 8.4, 8.0 Hz, 1H), 7.17-7.07 (m, 2H), 6.77 (dd, component of ABC pattern, J = 7.8, 7.8 Hz, 1H), 6.72-6.67 (m, 2H), 4.21 (q, J = 7.1 Hz, 2H), 3.27 (s, 2H), 3.07 (m, 2H), 2.70 (tt, J = 12.1, 3.8 Hz, 1H), 2.35 (ddd, J = 11.5, 11.5, 2.7 Hz, 2H), 2.04 (d, J = 1.1 Hz, 3H), 2.02-1.76 (m, 4H), 1.29 (t, J = 7.1 Hz, 3H).

[0587] Step 3: Synthesis of {4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}acetic acid (P10)

[0588] A solution of C14 (3.57 g, 8.23 ​​mmol) and aqueous sodium hydroxide solution (3 M; 13.7 mL, 41.1 mmol) in a mixture of methanol (80 mL) and tetrahydrofuran (40 mL) was stirred at 25 °C for 16 hours. After the solvent was removed in vacuo, the aqueous residue was acidified to pH 7 by adding 1 M hydrochloric acid and then extracted with a mixture of dichloromethane and methanol (10:1, 2 x 100 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure to provide P10 as a yellow solid. Yield: 2.95 g, 7.27 mmol, 88%. LCMS m / z 406.2◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4)δ7.61(dd, J=8.3,8.3Hz,1H),7.29(dd, J=10.9,2.0Hz,1H),7.22(ddd, J=8.4,2.0,0.8Hz,1H),6.82(dd, component of ABC pattern, J=8.3,7.1Hz,1H),6.78-6.72(m,2H),3.65-3.54(br m,2H),3.51(s,2H),3.04-2.88(m,3H),2.23-2.07(m,2H),2.07-1.93(m,2H),2.04(d,J=1.1Hz,3H).

[0589] Preparation of P11

[0590] 2-(Chloromethyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid methyl ester (P11)

[0591]

[0592] Step 1: Synthesis of methyl 3-[(2-methoxyethyl)amino]-4-nitrobenzoate (C15)

[0593] Triethylamine (40.7 g, 402 mmol, 55.8 mL) was added to a colorless solution of methyl 3-fluoro-4-nitrobenzoate (50 g, 250 mmol) in tetrahydrofuran (400 mL), and then 2-methoxyethylamine (30.2 g, 402 mmol) in tetrahydrofuran (100 mL) was added dropwise at room temperature. The resulting yellow solution was stirred for 18 hours at 55 ° C. The solution was cooled to room temperature and concentrated under reduced pressure to remove tetrahydrofuran. The resulting yellow solid was dissolved in ethyl acetate (800 mL) and washed with saturated aqueous ammonium chloride solution (250 mL). The aqueous phase was separated and extracted with ethyl acetate (200 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (3 x 250 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain C15 (60.2 g, 94%) as a yellow solid. 1 H NMR (600 MHz, CHLOROFORM-d) δ 8.23 ​​(d, 1H), 8.17 (br s, 1H), 7.58 (d, 1H), 7.25 (dd, 1H), 3.95 (s, 3H), 3.69-3.73 (m, 2H), 3.56 (m, 2H), 3.45 (s, 3H); LCMS m / z 255.4 [M+H] + .

[0594] Step 2: Synthesis of methyl 4-amino-3-[(2-methoxyethyl)amino]benzoate (C16)

[0595] Pd / C (10 g, 94 mmol) was added to a solution of C15 (30 g, 118 mmol) in methanol (500 mL). The reaction was stirred for 18 hours at room temperature under 15 psi hydrogen. The black suspension was filtered through celite and the filter cake was washed with methanol (500 mL). The combined filtrate was concentrated in vacuo to give C16 (26.5 g, quantitative) as a brown oil which solidified upon standing. 1 H NMR (400 MHz, CHLOROFORM-d) δ7.48 (dd, 1H), 7.36 (d, 1H), 6.69 (d, 1H), 3.87 (s, 3H), 3.77 (br s, 2H), 3.68 (t, 2H), 3.41 (s, 3H), 3.32 (t, 2H); LCMS m / z 224.7 [M+H] + .

[0596] Step 3: Synthesis of methyl 2-(chloromethyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (P11)

[0597] 2-Chloro-1,1,1-trimethoxyethane (3.31 mL, 24.6 mmol) and p-toluenesulfonic acid monohydrate (84.8 mg, 0.446 mmol) were added to a solution of C16 (5.00 g, 22.3 mmol) in tetrahydrofuran (100 mL). The reaction mixture was heated at 45 °C for 5 hours, whereupon it was concentrated in vacuo; the residual oil was dissolved in ethyl acetate (10 mL) and heated until a solution formed. It was stirred slowly while cooling to room temperature overnight. The precipitate was collected by filtration and washed with heptane to give P11 as a grey solid. Yield: 5.73 g, 20.3 mmol, 91%. 1 HNMR(600MHz,chloroform-d)δ8.12(br s,1H),8.01(br d,J=8.6Hz,1H),7.79(d,J=8.4Hz,1H),4.96(s,2H),4.52(t,J=5.1Hz,2H),3.96(s,3H),3.74(t,J=5.1Hz,2H),3.28(s,3H).

[0598] Step 4: Synthesis of methyl 2-(chloromethyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate, hydrochloride (P11, HCl salt)

[0599] A solution of C16 (5.0 g, 24 mmol) in 1,4-dioxane (100 mL) was heated to 100 ° C, a solution of chloroacetic anhydride (4.1 g, 24.5 mmol) in 1,4-dioxane (60 mL) was added via an addition funnel over a 10 hour period, and the reaction mixture was stirred for another 12 hours at 100 ° C. The next day, the reaction was cooled to room temperature and 1,4-dioxane was removed under reduced pressure. The crude reaction mixture was dissolved in ethyl acetate and washed with saturated aqueous sodium bicarbonate solution. The ethyl acetate layer was separated, dried over sodium sulfate, and filtered. Under constant stirring, a solution of 4M hydrogen chloride in 1,4-dioxane (1.1 equivalents) was added to the ethyl acetate solution. The hydrochloride salt of P11 precipitated out as a pale yellow solid. The suspension was stirred for 1 hour, and the hydrochloride salt of P11 was then collected by filtration to give a yellow solid (6.1 g, 86%). 1 H NMR (600MHz, CD 3 OD)δ8.64(s,1H),8.30(d,1H),7.92(d,1H),5.32(s,2H),4.84(m,2H),3.99(s,3H),3.83(t,2H),3.31(s,3H). LCMS m / z 283.2[M+H] + .

[0600] Preparation of P12

[0601] 1-(2-methoxyethyl)-2-(piperazin-1-ylmethyl)-1H-benzimidazole-6-carboxylic acid methyl ester (P12)

[0602]

[0603] Step 1: Synthesis of methyl 2-{[4-(tert-butoxycarbonyl)piperazin-1-yl]methyl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (C17)

[0604] Compound P11 (1.59 g, 5.62 mmol) was added to a 15° C. mixture of tert-butyl piperazine-1-carboxylate (1.00 g, 5.37 mmol) and potassium carbonate (2.97 g, 21.5 mmol) in acetonitrile (15 mL), and the reaction mixture was stirred at 55° C. for 12 hours. It was then combined with a similar reaction using P11 and tert-butyl piperazine-1-carboxylate (200 mg, 1.07 mmol), and the mixture was filtered. After the filtrate was concentrated in vacuo, the residue was purified via silica gel chromatography (gradient: 0% to 60% ethyl acetate in petroleum ether) to provide C17 as a light yellow solid. Combined yield: 2.30 g, 5.32 mmol, 83%. LCMS m / z 433.0 [M+H] + . 1 H NMR (400MHz, chloroform-d) δ8.12 (d, J=1.5Hz, 1H), 7.96 (dd, J=8.4, 1.5Hz, 1H), 7.73 (d, J=8.5Hz, 1H),4.58(t,J=5.4Hz,2H),3.95(s,3H),3.89(s,2H),3.73(t,J=5.4Hz,2H),3.46-3.37(br m,4H),3.28(s,3H),2.54-2.44(br m,4H),1.45(s,9H).

[0605] Step 2: Synthesis of methyl 1-(2-methoxyethyl)-2-(piperazin-1-ylmethyl)-1H-benzimidazole-6-carboxylate (P12)

[0606] A solution of hydrogen chloride in ethyl acetate (20 mL) was added to a solution of C17 (2.30 g, 5.32 mmol) in dichloromethane (80 mL). The reaction mixture was stirred at 20 °C for 2 hours, whereupon it was concentrated in vacuo. The residue was diluted with water (20 mL), the pH was adjusted to 9-10 by the addition of saturated aqueous sodium bicarbonate solution, and extracted with a mixture of ethyl acetate and methanol (10:1, 15 x 50 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to give P12 as a light yellow solid. Yield: 1.68 g, 5.05 mmol, 95%. LCMS m / z 332.8 [M+H] + . 1HNMR(400MHz,chloroform-d)δ8.13(br s,1H),7.96(br d,J=8.5Hz,1H),7.72(d,J=8.5Hz,1H),4.59(t,J=5.5Hz,2H),3.95(s,3H),3. 86(s,2H),3.75(t,J=5.5Hz,2H),3.29(s,3H),2.87(t,J=4.8Hz,4H),2.50(br m,4H).

[0607] Preparation of P13

[0608] 6-Bromo-2-(chloromethyl)-1-(2-methoxyethyl)-1H-imidazo[4,5-b]pyridine (P13)

[0609]

[0610] Step 1: Synthesis of 5-bromo-N-(2-methoxyethyl)-2-nitropyridin-3-amine (C18)

[0611] A solution of 5-bromo-3-fluoro-2-nitropyridine (400 mg, 1.81 mmol) and 2-methoxyethylamine (408 mg, 5.43 mmol) in tetrahydrofuran (10 mL) was stirred at 25° C. for 2 hours, whereupon it was diluted with ethyl acetate (100 mL) and washed with water (50 mL). The organic layer was washed with saturated aqueous sodium chloride solution (50 mL), dried over magnesium sulfate, filtered and concentrated to afford C18 as a yellow solid. Yield: 430 mg, 1.56 mmol, 86%.

[0612] Step 2: 5-Bromo-N 3 Synthesis of -(2-methoxyethyl)pyridine-2,3-diamine (C19)

[0613] A solution of C18 (430 mg, 1.56 mmol), ammonium chloride (833 mg, 15.6 mmol), and iron powder (870 mg, 15.6 mmol) in a mixture of methanol (10 mL) and water (2 mL) was stirred at 80° C. for 30 minutes. The resulting suspension was poured into water (50 mL) and extracted with ethyl acetate (2×50 mL); the combined organic layers were dried over magnesium sulfate, filtered, and concentrated to provide C19 as a brown solid. Yield: 350 mg, 1.42 mmol, 91%. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.63 (d, J = 2.1 Hz, 1H), 6.88 (d, J = 2.0 Hz, 1H), 4.33-4.19 (br s, 2H), 3.65 (dd, J = 5.6, 4.6 Hz, 2H), 3.40 (s, 3H), 3.22 (br t, J = 5 Hz, 2H).

[0614] Step 3: Synthesis of 6-bromo-2-(chloromethyl)-1-(2-methoxyethyl)-1H-imidazo[4,5-b]pyridine (P13)

[0615] A solution of C19 (400 mg, 1.63 mmol) in 1,4-dioxane (8 mL) was treated with chloroacetyl chloride (0.284 mL, 3.57 mmol) and stirred at room temperature until LCMS analysis showed complete conversion of C19 to the intermediate amide. After removal of 1,4-dioxane in vacuo, the residue was dissolved in trifluoroacetic acid (8 mL) and heated at 80 ° C for 18 hours, whereupon the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The resulting oil was dissolved in ethyl acetate (50 mL) and neutralized by addition of saturated aqueous sodium bicarbonate solution. The aqueous layer was extracted with ethyl acetate (20 mL), and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Silica gel chromatography (gradient: 0% to 80% ethyl acetate (in heptane)) gave P13 as a solid. Yield: 176 mg, 0.578 mmol, 35%. LCMS m / z 306.1 (Br-Cl isotope pattern observed) [M+H] + . 1 H NMR (600 MHz, CHLOROFORM-d) δ 8.58 (br s, 1H), 7.89 (br s, 1H), 4.92 (s, 2H), 4.44 (t, J = 5.0 Hz, 2H), 3.71 (t, J = 5.0 Hz, 2H), 3.28 (s, 3H).

[0616] Preparation of P14

[0617] 2-{[4-(2,3-dihydroxyphenyl)piperidin-1-yl]methyl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid methyl ester (P14)

[0618]

[0619]

[0620] Step 1: Synthesis of [(3-bromobenzene-1,2-diyl)bis(oxymethanediyloxyethane-2,1-diyl)]bis(trimethylsilane) (C20)

[0621] This reaction was carried out in two batches of the same scale. N,N-diisopropylethylamine (37.8 mL, 217 mmol) was added dropwise to a solution of 3-bromobenzene-1,2-diol (10.0 g, 52.9 mmol) in tetrahydrofuran (300 mL). After the mixture was stirred at 20°C for 10 minutes, [2-(chloromethoxy)ethyl](trimethyl)silane (19.2 mL, 108 mmol) was added dropwise over 5 minutes, and stirring was continued at room temperature (18°C) for 16 hours. N,N-diisopropylethylamine (27.6 mL, 158 mmol) was added again, followed by [2-(chloromethoxy)ethyl](trimethyl)silane (14.0 mL, 79.1 mmol) dropwise at room temperature (18°C). After an additional 2.5 hours at room temperature, the reaction mixture was filtered, and the filtrate was concentrated in vacuo. At this point, the crude product from both batches was combined and purified using silica gel chromatography (Gradient: 0% to 7% ethyl acetate in petroleum ether) to afford C20 as a colorless oil. 1 H NMR analysis showed that this material was not completely pure. Combined yield: 22.9 g, 50.9 mmol, 48%. 1 H NMR (400 MHz, chloroform-d), C20 peak only: δ 7.19 (dd, J = 8.1, 1.5 Hz, 1H), 7.12 (dd, J = 8.3, 1.4 Hz, 1H), 6.90 (dd, J = 8.2.8.2 Hz, 1H), 5.26-5.19 (m, 4H), 4.00-3.92 (m, 2H), 3.80-3.73 (m, 2H), 1.00-0.91 (m, 4H), 0.03 (s, 9H), 0.00 (s, 9H).

[0622] Step 2: Synthesis of tert-butyl 4-(2,3-bis{[2-(trimethylsilyl)ethoxy]methoxy}phenyl)-3,6-dihydropyridine-1(2H)-carboxylate (C21)

[0623] A reaction vessel containing C20 (6.11 g, 13.6 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (5.04 g, 16.3 mmol), aqueous sodium carbonate (1 M; 40.8 mL, 40.8 mmol), and a suspension of [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (497 mg, 0.679 mmol) in 1,4-dioxane (100 mL) was evacuated and filled with nitrogen. This evacuation cycle was repeated twice, then the reaction mixture was stirred at 85° C. for 16 hours, whereupon the reaction mixture was diluted with water (40 mL) and extracted with ethyl acetate (3×150 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Purification via silica gel chromatography (Gradient: 0% to 8% methanol in dichloromethane) provided C21 as a yellow oil. Yield: 5.47 g, 9.91 mmol, 73%. 1 H NMR (600MHz, chloroform-d) δ7.10(br d,J=8.2Hz,1H),6.98(dd,J=7.9,7.9Hz,1H),6.81(br d,J=7.7Hz,1H),5.79(br s,1H),5.23(s,2H),5.07(s,2H),4.03(brs,2H),3.83-3.74(m,4H),3.59(br s,2H),2.52(br s,2H),1.49(s,9H),1.01-0.89(m,4H),0.01(s,9H),0.01(s,9H).

[0624] Step 3: Synthesis of tert-butyl 4-(2,3-bis{[2-(trimethylsilyl)ethoxy]methoxy}phenyl)piperidine-1-carboxylate (C22)

[0625] A solution of C21 (12.5 g, 22.6 mmol) in methanol (300 mL) was treated with 10% palladium on carbon (2.94 g, 2.76 mmol) and hydrogenated at 40 psi and 25 °C for 16 h. LCMS analysis at this point indicated conversion to the product: LCMS m / z 576.0 [M+Na + After filtering the reaction mixture and washing the filter cake with methanol (2 x 100 mL), the combined filtrates were concentrated in vacuo to give C22 as a colorless oil. Yield: 11.2 g, 20.1 mmol, 89%. 1HNMR (400MHz, chloroform-d) δ7.05-6.97(m,2H),6.83(dd,J=6.9,2.5Hz,1H),5.22(s,2H),5.13(s,2H),4.38-4.10(br m,2H),3.90-3.82(m,2H),3.81-3.73(m,2H),3.22(tt,J=12.2,3.5Hz,1H),2.79(br dd,J=12.8,12.8Hz,2H),1.78(br d,J=13Hz,2H),1.65-1.52(m,2H),1.48(s,9H),1.04-0.91(m,4H),0.03(s,9H),0.00(s,9H).

[0626] Step 4: Synthesis of 4-(2,3-bis{[2-(trimethylsilyl)ethoxy]methoxy}phenyl)piperidine (C23)

[0627] 2,6-Lutidine (2.39 g, 22.3 mmol) was added to a room temperature (15 °C) solution of C22 (7.23 g, 13.0 mmol) in dichloromethane (90 mL), followed by the dropwise addition of trimethylsilyl trifluoromethanesulfonate (3.80 g, 17.1 mmol). The reaction mixture was stirred at 15 °C for 16 hours, whereupon additional 2,6-Lutidine (909 mg, 8.48 mmol) and trimethylsilyl trifluoromethanesulfonate (1.45 g, 6.52 mmol) were added. After stirring at room temperature (15 °C) for an additional 5 hours, LCMS analysis of the reaction mixture showed the presence of the product: LCMS m / z 454.1 [M+H] + The reaction mixture was concentrated in vacuo, and the residue was washed sequentially with aqueous ammonium chloride solution (3 x 100 mL) and saturated aqueous sodium chloride solution (100 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give C23 (6.6 g) as a brown oil. This material was used directly in the next step.

[0628] Step 5: Synthesis of methyl 2-{[4-(2,3-bis{[2-(trimethylsilyl)ethoxy]methoxy}phenyl)piperidin-1-yl]methyl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (C24)

[0629] P11 (3.08 g, 10.9 mmol) was added to a solution of C23 (from the previous step; 6.6 g, ≤ 13 mmol) in acetonitrile (150 mL), followed by potassium carbonate (10.1 g, 73.1 mmol), and the reaction mixture was stirred at room temperature (15°C) for 16 hours. LCMS analysis at this point indicated the presence of the product: LCMS m / z 700.2 [M+H] + The reaction mixture was filtered and the filtrate was concentrated in vacuo; purification via silica gel chromatography (Gradient: 34% to 56% ethyl acetate in petroleum ether) gave C24 as a yellow oil. Yield: 5.4 g, 7.7 mmol, 59% (over two steps). 1 HNMR (400MHz, chloroform-d) δ8.16-8.12(m,1H),7.96(dd,J=8.5,1.5Hz,1H),7.73(d,J=8.5Hz,1H),7.04-6.96(m,2H),6.86(dd,J=6.7,2.6Hz,1H), 5.21(s,2H),5.12(s,2H),4.63(t,J=5.5Hz,2H),3.95(s,3H),3.93-3. 83(m,4H),3.80-3.72(m,4H),3.31(s,3H),3.17-3.06(m,1H),2.99(br d, J = 11.2 Hz, 2H), 2.35-2.22 (m, 2H), 1.81 (br d, half of the AB quartet, J = 12.6 Hz, 2H), 1.75-1.61 (m, 2H), 1.04-0.91 (m, 4H), 0.05 (s, 9H), -0.01 (s, 9H).

[0630] Step 6: Synthesis of methyl 2-{[4-(2,3-dihydroxyphenyl)piperidin-1-yl]methyl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (P14)

[0631] A solution of hydrogen chloride in 1,4-dioxane (4M; 96 mL, 384 mmol) was added to a room temperature (18°C) solution of C24 (6.40 g, 9.14 mmol) in 1,4-dioxane (120 mL). After the addition was complete, the reaction mixture was stirred at room temperature (18°C) for 16 hours, combined with a similar reaction performed using C24 (1.00 g, 1.43 mmol) and concentrated in vacuo. The residue was treated with a mixture of dichloromethane and methanol (20:1, 150 mL) and stirred at room temperature (18°C) for 1 hour, whereupon the solid (4.85 g) was collected via filtration. This material was treated with water (100 mL) and the mixture was adjusted to pH 7-8 by the addition of aqueous sodium bicarbonate, stirred at room temperature (18°C) for 30 minutes, and filtered. The filter cake was washed with water (2 x 20 mL), then mixed with methanol (100 mL) and concentrated in vacuo. The resulting material was treated with petroleum ether (100 mL) and stirred at room temperature (18° C.) for 30 minutes. After filtration, the filter cake was mixed with toluene (30 mL) and concentrated in vacuo to provide P14 as a gray solid. Combined yield: 2.92 g, 6.64 mmol, 63%. LCMS m / z 440.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ8.21(d,J=1.6Hz,1H),7.81(dd,J=8.5,1.6Hz,1H),7.66(d,J=8.5Hz,1H),6.64-6.51(m,3H),4.63(t, J=5.3Hz,2H),3.88(s,3H),3.84(s,2H),3.75(t,J=5.3Hz,2H),3.22(s,3H),2.97-2.78(m,3H),2.18(br dd,J=11,11Hz,2H),1.75-1.64(m,2H),1.64-1.49(m,2H).

[0632] Preparation of P15

[0633] 2-(Chloromethyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid methyl ester (P15)

[0634]

[0635] This whole sequence is carried out on a large scale. Generally speaking, before the reaction and after adding the reagents, the reactor is evacuated to -0.08 to -0.05 MPa and then filled to normal pressure with nitrogen. This process is usually repeated three times, and the oxygen content is then evaluated to ensure that it is ≤1.0%. For the extraction and washing process of the organic layer, the mixture is usually stirred for 15 to 60 minutes, and then allowed to stand for 15 to 60 minutes before the layers are separated.

[0636] Step 1: Synthesis of (2S)-2-[(benzyloxy)methyl]oxetane (C25)

[0637] This reaction is carried out in three batches of approximately the same scale. 2-methyl propan-2-ol (774.7kg) is loaded into a 2000L glass-lined reactor. Potassium tert-butoxide (157.3kg, 1402mol) is added via a solid addition funnel, and the mixture is stirred for 30 minutes. Then, trimethyl sulfoxide iodide (308.2kg, 1400mol) is added in the same manner, and at 55°C to 65°C, the reaction mixture is heated for 2 to 3 hours, then at a rate of 5 to 20kg / hour, (2S)-2-[(benzyloxy)methyl]oxirane (92.1kg, 561mol) is added. After the reaction mixture is maintained at 55°C to 65°C for 25 hours, it is cooled to 25°C to 35°C, and filtered through diatomaceous earth (18.4kg). The filter cake was rinsed with tert-butyl methyl ether (3 x 340 kg), and the combined filtrates were transferred to a 5000 L reactor, treated with pure water (921 kg), and stirred at 15° C. to 30° C. for 15 to 30 minutes. The organic layer was then washed twice with a solution of sodium chloride (230.4 kg) in pure water (920.5 kg), and concentrated under reduced pressure (≤-0.08 MPa), ≤45° C. n-heptane (187 kg) was added, and the resulting mixture was concentrated under reduced pressure (≤-0.08 MPa), ≤45° C.; the organic phase was purified using silica gel chromatography (280 kg) with sodium chloride (18.5 kg) at the top of the column. The crude was loaded into the column using n-heptane (513 kg), and then eluted with a mixture of n-heptane (688.7 kg) and ethyl acetate (64.4 kg). The three batches were combined to afford C25 (189.7 kg, 906 mmol, 54%) as a 85% pure light yellow oil. 1H NMR (400 MHz, chloroform-d), C25 peak only: δ 7.40-7.32 (m, 4H), 7.32-7.27 (m, 1H), 4.98 (dddd, J = 8.1, 6.7, 4.9, 3.7 Hz, 1H), 4.72-4.55 (m, 4H), 3.67 (dd, component of ABX pattern, J = 11.0, 4.9 Hz, 1H), 3.62 (dd, component of ABX pattern, J = 11.0, 3.7 Hz, 1H), 2.72-2.53 (m, 2H).

[0638] Step 2: Synthesis of (2S)-oxetan-2-ylmethanol (C26)

[0639] 10% palladium on carbon (30.7 kg) was added to a 10°C-30°C solution of 85% pure C25 (from the previous step, 185.3 kg, 884.8 mol) in tetrahydrofuran (1270 kg) in a 3000 L stainless steel autoclave reactor via an addition funnel. The addition funnel was rinsed with pure water and tetrahydrofuran (143 kg), and the rinse was added to the reaction mixture. After the reactor contents were purged with nitrogen, they were similarly purged with hydrogen, the pressure was increased to 0.3 to 0.5 MPa, and then vented to 0.05 MPa. The hydrogen purge was repeated five times, whereupon the hydrogen pressure was increased to 0.3 to 0.4 MPa. The reaction mixture was then heated to 35°C-45°C. After 13 hours, the mixture was vented to 0.05 MPa while maintaining the hydrogen pressure at 0.3 to 0.5 MPa, and purged with nitrogen five times by increasing the pressure to 0.15-0.2 MPa and then venting to 0.05 MPa. After the mixture was cooled to 10° C. to 25° C., it was filtered and the reactor was rinsed with tetrahydrofuran (2 x 321 kg). The filter cake was soaked twice with this rinse and then filtered; concentrated under reduced pressure (≤-0.06 MPa) at ≤40° C. to give C26 (62.2 kg, 706 mol, 80%) in tetrahydrofuran (251 kg).

[0640] Step 3: Synthesis of (2S)-oxetan-2-ylmethyl 4-methylbenzenesulfonate (C27)

[0641] 4-(Dimethylamino)pyridine (17.5 kg, 143 mol) was added to a 10°C to 25°C solution of C26 (from the previous step; 62.2 kg, 706 mol) in tetrahydrofuran (251 kg) and triethylamine (92.7 kg, 916 mol in 1240 kg dichloromethane). After 30 minutes, p-toluenesulfonyl chloride (174.8 kg, 916.9 mol) was added portionwise at 20 to 40 minute intervals, and the reaction mixture was stirred at 15 to 25°C for 16 hours and 20 minutes. Pure water (190 kg) was added; after stirring, the organic layer was washed with an aqueous sodium bicarbonate solution (prepared using 53.8 kg sodium bicarbonate and 622 kg pure water), and then washed with an aqueous ammonium chloride solution (prepared using 230 kg ammonium chloride and 624 kg pure water). After a final wash with purified water (311 kg), the organic layer was filtered through a stainless steel Nutsche filter pre-packed with silica gel (60.2 kg). The filter cake was soaked with dichloromethane (311 kg) for 20 minutes and then filtered; the combined filtrate was concentrated under reduced pressure (≤-0.05 MPa) and ≤40°C until 330 to 400 L remained. Tetrahydrofuran (311 kg) was then added at 15-30°C, and the mixture was concentrated to a final volume of 330 to 400 L in the same manner. The addition of tetrahydrofuran and concentration were repeated, again to a volume of 330 to 400 L, to give a light yellow solution of C27 (167.6 kg, 692 mmol, 98%) in tetrahydrofuran (251.8 kg). 1 H NMR (400 MHz, chloroform-d), C27 peak only: δ 7.81 (d, J = 8.4 Hz, 2H), 7.34 (d, J = 8.1 Hz, 2H), 4.91 (ddt, J = 8.0, 6.7, 3.9 Hz, 1H), 4.62-4.55 (m, 1H), 4.53-4.45 (m, 1H), 4.14 (d, J = 3.9 Hz, 2H), 2.75-2.63 (m, 1H), 2.60-2.49 (m, 1H), 2.44 (s, 3H).

[0642] Step 4: Synthesis of (2S)-2-(azidomethyl)oxetane (C28)

[0643] N,N-dimethylformamide (473 kg), sodium azide (34.7 kg, 534 mol), and potassium iodide (5.2 kg, 31 mol) were combined in a 3000 L glass-lined reactor at 10° C. to 25° C. After adding C27 (83.5 kg, 344.6 mol, in 125.4 kg tetrahydrofuran), the reaction mixture was heated to 55° C. to 65° C. for 17 hours and 40 minutes, whereupon it was cooled to 25-35° C. and nitrogen was bubbled through the bottom valve for 15 minutes. Then, tert-butyl methyl ether (623 kg) and pure water (840 kg) were added, and the resulting aqueous layer was extracted twice with tert-butyl methyl ether (312 kg and 294 kg). The combined organic layers were washed with pure water (2 x 419 kg) while maintaining the temperature at 10° C. to 25° C. to obtain C28 (31.2 kg, 276 mol, 80%) in the aforementioned organic layer solution (1236.8 kg).

[0644] Step 5: Synthesis of 1-[(2S)-oxetan-2-yl]methanamine (C29)

[0645] 10% palladium on carbon (3.7 kg) was added to a 10°C to 30°C solution of C28 [from the previous step, 1264 kg (31.1 kg of C28, 275 mol)] in tetrahydrofuran (328 kg) in a 3000 L stainless steel autoclave reactor via an addition funnel. The addition funnel was rinsed with tetrahydrofuran (32 kg) and the rinse was added to the reaction mixture. After the reactor contents were purged with nitrogen, they were similarly purged with hydrogen, the pressure was increased to 0.05 to 0.15 MPa, and then vented to 0.03-0.04 MPa. This hydrogen purge was repeated five times, whereupon the hydrogen pressure was increased to 0.05 to 0.07 MPa. The reaction temperature was increased to 25°C-33°C, and the hydrogen pressure was maintained at 0.05-0.15 MPa for 22 hours, while the hydrogen was replaced every 3 to 5 hours. The mixture was then purged five times with nitrogen by increasing the pressure to 0.15-0.2 MPa and then venting to 0.05 MPa. After filtration, tetrahydrofuran (92 kg and 93 kg) was used to wash the reactor and then soak the filter cake. The combined filtrate was concentrated under reduced pressure (≤-0.07 MPa) and ≤45°C to give C29 (18.0 kg, 207 mol, 75%) in tetrahydrofuran (57.8 kg). 1 H NMR (400 MHz, DMSO-d 6), only C29 peak: δ4.62 (ddt, J = 7.6, 6.6, 5.1 Hz, 1H), 4.49 (ddd, J = 8.6, 7.3, 5.6 Hz, 1H), 4.37 (dt, J = 9.1, 5.9 Hz, 1H), 2.69 (d, J = 5.1 Hz, 2H), 2.55-2.49 (m, 1H), 2.39 (m, 1H).

[0646] Step 6: Synthesis of methyl 4-nitro-3-{[(2S)-oxetan-2-ylmethyl]amino}benzoate (C30)

[0647] Potassium carbonate (58.1 kg, 420 mol) was added to a solution of methyl 3-fluoro-4-nitrobenzoate (54.8 kg, 275 mol) in tetrahydrofuran (148 kg) in a 100 L glass-lined reactor, and the mixture was stirred for 10 minutes. A solution of C29 (29.3 kg, 336 mol) in tetrahydrofuran (212.9 kg) was added, and the reaction mixture was stirred for 12 hours at 20° C. to 30° C., whereupon ethyl acetate (151 kg) was added, and the mixture was filtered through silica gel (29 kg). The filter cake was rinsed with ethyl acetate (150 kg and 151 kg), and the combined filtrate was concentrated to a volume of 222-281 L at reduced pressure (≤−0.08 MPa) and ≤45° C. After the mixture was cooled to 10°C-30°C, n-heptane (189 kg) was added, stirring was performed for 20 minutes, and the mixture was concentrated to a volume of 222 L at reduced pressure (≤-0.08 MPa) and ≤45°C. n-heptane (181 kg) was added to the mixture again at a reference rate of 100 to 300 kg / hour, and stirring was continued for 20 minutes. The mixture was sampled until the residual tetrahydrofuran was ≤5% and the remaining ethyl acetate was 10% to 13%. The mixture was heated to 40°C-45°C and stirred for 1 hour, whereupon it was cooled to 15°C-25°C at a rate of 5°C to 10°C per hour, and then stirred at 15°C to 25°C for 1 hour. Filtration using a stainless steel centrifuge provided a filter cake, which was rinsed with a mixture of ethyl acetate (5.0 kg) and n-heptane (34 kg), and then stirred with tetrahydrofuran (724 kg) at 10-30° C. for 15 minutes; filtration provided a yellow solid consisting mostly of C30 (57.3 kg, 210 mol, 76%). 1 H NMR (400 MHz, DMSO-d 6)8.34(t,J=5.8Hz,1H),8.14(d,J=8.9Hz,1H),7.63(d,J=1.7Hz,1H),7.13(dd,J=8.9,1.8Hz,1H),4.99(dddd,J=7.7,6.7,5.3,4.1Hz,1H),4.55(dd d,J=8.6,7.3,5.8Hz,1H),4.43(dt,J=9.1,6.0Hz,1H),3.87(s,3H),3.67- 3.61(m,2H),2.67(dddd,J=11.1,8.6,7.7,6.2Hz,1H),2.57-2.47(m,1H).

[0648] Step 7: Synthesis of methyl 2-(chloromethyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (P15)

[0649] A solution of C30 (from the previous step, 51.8 kg, 190 mol) in tetrahydrofuran (678 kg) in a 3000 L autoclave reactor was treated with 10% palladium on carbon (5.2 kg) at 10°C to 30°C. The feed line was flushed with tetrahydrofuran (46 kg), and the flush was added to the reaction mixture. After the reactor contents were purged with nitrogen, they were similarly purged with hydrogen, the pressure was increased to 0.1 to 0.2 MPa, and then vented to 0.02-0.05 MPa. This hydrogen purge was repeated five times, whereupon the hydrogen pressure was increased to 0.1 to 0.25 MPa. The reaction mixture was stirred at 20°C-30°C, and every 2 to 3 hours, the mixture was purged three times with nitrogen, and then purged five times with hydrogen; after each final hydrogen exchange, the hydrogen pressure was increased to 0.1 to 0.25 MPa. After 11.25 hours total reaction time, the reaction mixture was vented to normal pressure and purged with nitrogen five times by increasing the pressure to 0.15-0.2 MPa and then venting to 0.05 MPa. It was then filtered and the filter cake was rinsed twice with tetrahydrofuran (64 kg and 63 kg); the combined rinse and filtrate were concentrated to a volume of 128-160 L at reduced pressure (≤-0.08 MPa) and ≤40° C. Tetrahydrofuran (169 kg) was added and the mixture was concentrated again to a volume of 128-160 L; this process was repeated a total of four times to give a solution of the intermediate methyl 4-amino-3-{[(2S)-oxetan-2-ylmethyl]amino}benzoate.

[0650] Tetrahydrofuran (150 kg) is added to this solution, followed by 2-chloro-1,1,1-trimethoxyethane (35.1 kg, 227 mol) and p-toluenesulfonic acid monohydrate (1.8 kg, 9.5 mol). After stirring the reaction mixture for 25 minutes, it is heated at 40°C to 45°C for 5 hours, whereupon it is concentrated to a volume of 135 to 181 L under reduced pressure. 2-Propanol (142 kg) is added, and the mixture is concentrated again to a volume of 135-181 L, whereupon 2-propanol (36.5 kg) and pure water (90 kg) are added, and stirring is continued until a solution is obtained. It is filtered using an in-line liquid filter, and then, at 20°C to 40°C, at a reference rate of 150 to 400 kg / hour, it is treated with pure water (447 kg). After the mixture was cooled to 20-30°C, it was stirred for 2 hours and the solid was collected by filtration using a centrifuge. The filter cake was rinsed with a solution of 2-propanol (20.5 kg) and purified water (154 kg); after drying, P15 (32.1 kg, 109 mol, 57%) was obtained as a white solid. 1 H NMR (400 MHz, chloroform-d) δ 8.14-8.11 (m, 1H), 8.01 (dd, J = 8.5, 1.1 Hz, 1H), 7.79 (br d, J = 8.6 Hz, 1H), 5.26-5.18 (m, 1H), 5.04 (s, 2H), 4.66-4.58 (m, 2H), 4.53 (dd, component of ABX pattern, J = 15.7, 2.7 Hz, 1H), 4.34 (dt, J = 9.1, 6.0 Hz, 1H), 3.96 (s, 3H), 2.82-2.71 (m, 1H), 2.48-2.37 (m, 1H).

[0651] Preparation of P16

[0652] 2-(Chloromethyl)-1-methyl-1H-benzimidazole-6-carboxylic acid methyl ester (P16)

[0653]

[0654] Methyl 4-amino-3-(methylamino)benzoate (206 mg, 1.14 mmol) was dissolved in 1,4-dioxane (11.5 mL) and treated with chloroacetyl chloride (109 μL, 1.37 mmol). The mixture was stirred at 100 °C for 3 hours and cooled to room temperature. Triethylamine (0.8 mL, 7 mmol) and heptane (10 mL) were added and filtered. The filtrate was concentrated under reduced pressure and the crude was purified by silica gel chromatography (eluent: 40% ethyl acetate in heptane) to give 120 mg of P16 (44%).1 H NMR (400 MHz, chloroform-d) δ8.14 (s, 1H), 8.01 (d, 1H), 7.78 (d, 1H), 4.87 (s, 2H), 3.97 (s, 3H), 3.94 (s, 3H); LCMS m / z 239.1 [M+H] + .

[0655] Preparation of P17 and P18

[0656] 2-(6-Azaspiro[2.5]octan-1-yl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid methyl ester, ENT-1(P17) and 2-(6-Azaspiro[2.5]octan-1-yl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid methyl ester, ENT-2(P18)

[0657]

[0658] Step 1: Synthesis of tert-butyl 4-(2-ethoxy-2-oxoethylidene)piperidine-1-carboxylate (C31)

[0659] A solution of potassium tert-butoxide (65.9 g, 587 mmol) in tetrahydrofuran (500 mL) was added to a 0°C solution of ethyl (diethoxyphosphoryl)acetate (132 g, 589 mmol) in tetrahydrofuran (500 mL), and the resulting suspension was stirred at 0°C for 1 hour, whereupon it was cooled to -50°C. A solution of tert-butyl 4-oxopiperidine-1-carboxylate (90.0 g, 452 mmol) in tetrahydrofuran (1.5 L) was added dropwise at -50°C, and the reaction mixture was then slowly warmed to 20°C and then stirred at 20°C for 16 hours. After adding water (1 L), the mixture was concentrated in vacuo to remove tetrahydrofuran. The aqueous residue was extracted with ethyl acetate (2 x 800 mL), and the combined organic layers were washed with saturated aqueous sodium chloride solution (500 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting material was washed several times with petroleum ether (200 mL) to provide C31 as a white solid. Yield: 95.0 g, 353 mmol, 78%. 1 H NMR (400 MHz, CHLOROFORM-d) δ 5.71 (s, 1H), 4.16 (q, J = 7.2 Hz, 2H), 3.55-3.43 (m, 4H), 2.94 (br t, J = 5.5 Hz, 2H), 2.28 (brt, J = 5.5 Hz, 2H), 1.47 (s, 9H), 1.28 (t, J = 7.0 Hz, 3H).

[0660] Step 2: Synthesis of 6-tert-butyl 1-ethyl 6-azaspiro[2.5]octane-1,6-dicarboxylate (C32)

[0661] Potassium tert-butoxide (71.2 g, 634 mmol) was added in one portion to a solution of trimethylsulfoxide iodide (140 g, 636 mol) in dimethylsulfoxide (800 mL) at 20°C. After the reaction mixture was stirred at 20°C for 1.5 hours, a solution of C31 (95.0 g, 353 mmol) in dimethylsulfoxide (800 mL) was added dropwise, and stirring was continued at 20°C for 16 hours. Then, saturated aqueous sodium chloride solution (2.0 L) was added; the resulting mixture was neutralized by adding ammonium chloride and extracted with ethyl acetate (3.0 L). The combined organic layers were washed successively with water (2 x 1.0 L) and saturated aqueous sodium chloride solution (2.0 L), dried over sodium sulfate, filtered, and concentrated in vacuo. Purification was performed via silica gel chromatography (eluent: 10:1 petroleum ether / ethyl acetate) to give C32 as a yellow oil. 1 H NMR analysis indicated the presence of additional aliphatic material. Yield: 80 g, 280 mmol, 79%. 1 H NMR (400 MHz, chloroform-d), C32 peak only: δ 4.19-4.09 (m, 2H), 3.55-3.39 (m, 3H), 3.27 (ddd, J = 13.0, 7.0, 4.5 Hz, 1H), 1.76-1.64 (m, 2H), 1.56 (dd, J = 8.0, 5.5 Hz, 1H, assumed; partially obscured by water peak), 1.47 (s, 9H), 1.47-1.37 (m, 2H), 1.27 (t, J = 7.0 Hz, 3H), 1.17 (dd, J = 5.0, 5.0 Hz, 1H), 0.93 (dd, J = 8.0, 4.5 Hz, 1H).

[0662] Step 3: Synthesis of 6-(tert-butoxycarbonyl)-6-azaspiro[2.5]octane-1-carboxylic acid (C33)

[0663] Lithium hydroxide monohydrate (37.4 g, 891 mmol) was added once to a mixture of C32 (80 g, 280 mmol) in tetrahydrofuran (500 mL) and water (500 mL). The reaction mixture was stirred for 16 hours at 25 ° C, whereupon it was diluted with water (600 mL) and washed with ethyl acetate (3 x 300 mL). The organic layer was discarded, and the aqueous layer was acidified to pH 3-4 by adding 6M hydrochloric acid. The resulting mixture was extracted with ethyl acetate (3 x 600 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was triturated with petroleum ether (300 mL) to provide C33 as a white solid. Yield: 42.0 g, 164 mmol, 59%. LCMS m / z 278.2 [M+Na + ]. 1 HNMR (400 MHz, DMSO-d 6 )δ12.15-12.03 (br s, 1H), 3.43-3.25 (m, 3H, assumed; partially obscured by water peak), 3.23-3.12 (m, 1H), 1.64-1.50 (m, 2H), 1.52 (dd, J=7.5, 5.5 Hz, 1H), 1.39 (s, 9H), 1.39-1.28 (m, 2H), 0.96-0.88 (m, 2H).

[0664] Step 4: Synthesis of tert-butyl 1-({4-(methoxycarbonyl)-2-[(2-methoxyethyl)amino]phenyl}carbamoyl)-6-azaspiro[2.5]octane-6-carboxylate (C34)

[0665] A solution of C33 (570 mg, 2.23 mmol), C16 (500 mg, 2.23 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 1.27 g, 3.34 mmol) in N,N-dimethylformamide (10 mL) was stirred at 30 °C for 30 minutes, whereupon triethylamine (902 mg, 8.91 mmol) was added and stirring was continued at 30 °C for 16 hours. The reaction mixture was then poured into water (60 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (3 x 50 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (eluent: 1:1 petroleum ether / ethyl acetate) afforded C34 as a brown oil, which was used directly in the next step.

[0666] Step 5: Synthesis of methyl 2-[6-(tert-butoxycarbonyl)-6-azaspiro[2.5]octan-1-yl]-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (C35)

[0667] A solution of C34 (from the previous step, ≤ 2.23 mmol) in acetic acid (15 mL) was stirred at 50 °C for 16 h, whereupon it was concentrated in vacuo to provide C35 as a brown oil. This material was used directly in the next step. LCMS m / z 444.1 [M+H] + .

[0668] Step 6: Synthesis of methyl 2-(6-azaspiro[2.5]octan-1-yl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (C36)

[0669] Trifluoroacetic acid (5 mL) was added to a solution of C35 (from the previous step; ≤ 2.23 mmol) in dichloromethane (10 mL), and the reaction mixture was stirred at 25 °C for 2 hours. After removal of the solvent in vacuo, the residue was basified by addition of saturated aqueous potassium carbonate (40 mL) and extracted with a mixture of dichloromethane and methanol (10:1, 3x40 mL). The combined organic layers were dried over magnesium sulfate, filtered, concentrated in vacuo, and subjected to silica gel chromatography (eluent: 10:1:0.1 dichloromethane / methanol / concentrated ammonium hydroxide) to give C36 as a yellow solid. Yield: 640 mg, 1.86 mmol, 83% (over three steps). LCMS m / z 344.1 [M+H] + .

[0670] Step 7: Separation of methyl 2-(6-azaspiro[2.5]octan-1-yl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate, ENT-1 (P17) and methyl 2-(6-azaspiro[2.5]octan-1-yl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate, ENT-2 (P18)

[0671] C36 (630 mg, 1.83 mmol) was separated into its constituent enantiomers using SFC [column: Chiral Technologies Chiralpak AD, 10 μm; mobile phase: 55:45 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting peak was designated as ENT-1 (P17), and the second eluting enantiomer was designated as ENT-2 (P18); both were isolated as pale yellow solids.

[0672] P17 yield: 300 mg, 0.874 mmol, 48%. LCMS m / z 344.1 [M+H] + Retention time: 5.10 min (column: Chiral Technologies Chiralpak AD-3, 4.6 x 150 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: ethanol containing 0.05% diethylamine; gradient: 5% to 40% B over 5.5 min, then hold at 40% B for 3.0 min; flow rate: 2.5 mL / min).

[0673] P18 yield: 240 mg, 0.699 mmol, 38%. LCMS m / z 344.1 [M+H] + Retention time: 7.35 minutes (analytical conditions were the same as those used for P17).

[0674] Preparation of P19

[0675] Methyl 4-amino-3-{[(1-ethyl-1H-imidazol-5-yl)methyl]amino}benzoate (P19)

[0676]

[0677] Step 1: Synthesis of methyl 3-{[(1-ethyl-1H-imidazol-5-yl)methyl]amino}-4-nitrobenzoate (C37)

[0678] Triethylamine (3.65 mL, 26.2 mmol) was added to a solution of methyl 3-fluoro-4-nitrobenzoate (1.00 g, 5.02 mmol) and 1-(1-ethyl-1H-imidazol-5-yl)methanamine, dihydrochloride (1.00 g, 5.05 mmol) in a mixture of tetrahydrofuran (12 mL) and methanol (8 mL). The reaction mixture was stirred at 60 °C for 40 hours, whereupon it was concentrated in vacuo and purified using silica gel chromatography (Gradient: 0% to 2% methanol in dichloromethane) to afford C37 as an orange solid. Yield: 1.27 g, 4.17 mmol, 83%. 1 H NMR (400MHz, chloroform-d) δ8.24(d,J=8.8Hz,1H),7.98-7.91(m,1H),7.68(d,J=1.7Hz,1H),7.57(br s,1H),7.33(dd,J=8.8,1.7Hz,1H),7.11(br s, 1H), 4.53 (d, J = 4.9Hz, 2H), 3.99 (q, J = 7.3Hz, 2H), 3.95 (s, 3H), 1.47 (t, J = 7.3Hz, 3H).

[0679] Step 2: Synthesis of methyl 4-amino-3-{[(1-ethyl-1H-imidazol-5-yl)methyl]amino}benzoate (P19)

[0680] A mixture of wet palladium on charcoal (144 mg) and C37 (412 mg, 1.35 mmol) in methanol (13 mL) was stirred under a hydrogen balloon at 25° C. for 16 hours. The reaction mixture was then filtered through a pad of celite and the filtrate was concentrated in vacuo to give P19 as a grey solid. Yield: 340 mg, 1.24 mmol, 92%. 1 H NMR (400 MHz, methanol-d 4 )δ7.66(br s,1H),7.38-7.29(m,2H),6.97(br s, 1H), 6.67 (d, J = 7.9Hz, 1H), 4.35 (s, 2H), 4.11 (q, J = 7.3Hz, 2H), 3.81 (s, 3H), 1.44 (t, J = 7.3Hz, 3H).

[0681] Preparation of P20

[0682] 4-Amino-3-(methylamino)benzoic acid methyl ester (P20)

[0683]

[0684] Step 1: Synthesis of methyl 3-(methylamino)-4-nitrobenzoate (D1)

[0685] Over 10 minutes, methylamine (38.4 mL, 76.8 mmol, 2 M in tetrahydrofuran) was added dropwise to a solution of methyl 3-fluoro-4-nitrobenzoate (5.10 g, 25.6 mmol) in tetrahydrofuran (60 mL). After addition, the pale yellow solution immediately turned dark orange and was stirred at room temperature for 2 hours. The reaction mixture was then diluted with ether (100 mL) and the organic layer was washed successively with water (50 mL) and saturated aqueous sodium chloride solution (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give 5.26 g of methyl 3-(methylamino)-4-nitrobenzoate (98%) as a dark orange solid. LCMS m / z 211.1 [M+H] + . 1H NMR (400 MHz, CHLOROFORM-d) δ 8.22 (d, J = 8.9 Hz, 1H), 8.00 (brs, 1H), 7.56 (d, J = 1.7 Hz, 1H), 7.25 (dd, J = 8.9, 1.7 Hz, 1H, assumed; partially obscured by solvent peak), 3.95 (s, 3H), 3.09 (d, J = 5.1 Hz, 3H).

[0686] Step 2: Synthesis of methyl 4-amino-3-(methylamino)benzoate (P20)

[0687] A solution of D1 (5.26 g, 25.0 mmol) in ethanol (150 mL) was added to a 500 mL flask pre-charged with 10% palladium on carbon (50% water; 1 g). The mixture was shaken at room temperature under 50 psi hydrogen atmosphere for 1 hour, whereupon it was filtered and the filter cake was rinsed with ethanol (100 mL). The filtrate was concentrated under reduced pressure to give 4.38 g of P20 (97%) as an off-white solid. LCMS m / z 181.1 [M+H] + . 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.46 (dd, J = 8.0, 1.9 Hz, 1H), 7.34 (d, J = 1.8 Hz, 1H), 6.68 (d, J = 8.0 Hz, 1H), 3.87 (s, 3H), 3.72 (br s, 2H), 3.21 (br s, 1H), 2.91 (s, 3H).

[0688] Preparation of P21 and P22

[0689] 5-Bromo-N 3 -methylpyridine-2,3-diamine (P21) and 5-bromo-N 3 ,6-Dimethylpyridine-2,3-diamine (P22)

[0690]

[0691] Intermediate P21 was synthesized according to literature procedure (Choi, JY et al., J. Med. Chem. 2012, 55, 852-870). Intermediate P22 was synthesized using the same method.

[0692] Preparation of P23

[0693] 2-(Chloromethyl)-1-[(1-methyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid methyl ester (P23)

[0694]

[0695] Step 1: Synthesis of methyl 3-{[(1-methyl-1H-imidazol-5-yl)methyl]amino}-4-nitrobenzoate (D2)

[0696] 1-(1-Methyl-1H-imidazol-5-yl)methanamine (670 mg, 6.0 mmol) and triethylamine (762 mg, 7.53 mmol) were slowly added to a colorless solution of methyl 3-fluoro-4-nitrobenzoate (1.0 g, 5.0 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was stirred at 60 °C for 16 hours, whereupon it was poured into water (30 mL) and extracted with dichloromethane (3 x 30 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by silica gel chromatography (eluent: 20% methanol in dichloromethane). The resulting yellow solid was triturated with 30:1 petroleum ether / ethyl acetate to give D2 (1.2 g, 82%) as a yellow solid. LCMS m / z 290.9 [M+H] + . 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.25 (d, J = 8.9 Hz, 1H), 7.98-7.92 (m, 1H), 7.70 (d, J = 1.7 Hz, 1H), 7.49 (s, 1H), 7.34 (dd, J = 8.9, 1.7 Hz, 1H), 7.12 (s, 1H), 4.54 (d, J = 5.0 Hz, 2H), 3.96 (s, 3H), 3.67 (s, 3H).

[0697] Step 2: Synthesis of methyl 4-amino-3-{[(1-methyl-1H-imidazol-5-yl)methyl]amino}benzoate (D3)

[0698] Wet 10% palladium on carbon (1 g) was added to a suspension of D2 (5.46 g, 18.8 mmol) in methanol (160 mL). The mixture was stirred at 20 °C under 1 atmosphere of hydrogen for 36 hours. The reaction mixture was filtered and the filter cake was rinsed with methanol (200 mL). The filtrate was concentrated under reduced pressure to give D3 (4.8 g, 98%) as a brown solid. LCMS m / z 260.9 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ7.56(s,1H),7.18(br d,J=8.1Hz,1H),7.12(brs,1H),6.87(s,1H),6.55(d,J=8.2Hz,1H),5.50(s, 2H), 4.84 (t, J = 5.2Hz, 1H), 4.23 (d, J = 5.0Hz, 2H), 3.73 (s, 3H), 3.63 (s, 3H).

[0699] Step 3: Synthesis of methyl 2-(hydroxymethyl)-1-[(1-methyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylate (D4)

[0700] A mixture of D3 (780 mg, 3.00 mmol) and 2-hydroxyacetic acid (342 mg, 4.49 mmol) in 1,3,5-trimethylbenzene (8 mL) was stirred at 140 ° C for 14 hours and at 25 ° C for 48 hours. The clear yellow solution was decanted to give a brown residue, which was dissolved in methanol (50 mL) and concentrated under reduced pressure. The crude was purified by silica gel chromatography (eluent: 20% methanol (in dichloromethane)) to provide D4 (318 mg, 35%) as a yellow foam. LCMS m / z 300.9 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ8.13-8.11(m,1H),7.83(dd,J=8.4,1.6Hz,1H),7.71(d,J=8.5Hz,1H),7. 59(s,1H),6.58(s,1H),5.69(s,2H),4.75(s,2H),3.84(s,3H),3.53(s,3H).

[0701] Step 4: Synthesis of methyl 2-(chloromethyl)-1-[(1-methyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylate (P23)

[0702] Thionyl chloride (990 mg, 0.60 mL, 8.32 mmol) was added dropwise to a suspension of D4 (500 mg, 1.66 mmol) in dichloromethane (10 mL) and N,N-dimethylformamide (3 mL) at room temperature. The reaction mixture was stirred for 1 hour at room temperature and then concentrated under reduced pressure. The resulting brown residue was triturated with dichloromethane (10 mL). The solid was collected by filtration and rinsed with dichloromethane (5 mL) to provide P23 (431 mg, 73%) as an off-white solid. LCMS m / z 318.9 ◆ [M+H] + . 1H NMR (400 MHz, DMSO-d 6 )δ9.17(s,1H),8.31(s,1H),7.93(br d,J=8.5Hz,1H),7.82(d,J=8.5Hz,1H),7.11(s,1H),5.92(s,2H),5.13(s,2H),3.87(s,3H),3.87(s,3H).

[0703] Preparation of P24

[0704] 5-Chloro-2-(chloromethyl)-3-methyl-3H-imidazo[4,5-b]pyridine (P24)

[0705]

[0706] Step 1: Synthesis of 6-chloro-N-methyl-3-nitropyridin-2-amine (D5)

[0707] A solution of methylamine in tetrahydrofuran (2.0 M; 622 mL, 1.24 mol) was added dropwise to 2,6-dichloro-3-nitropyridine (200 g, 1.04 mol) and Na 2 CO 3 (132g, 1.24mol) in a suspension of ethanol (1L). After the addition was completed, the reaction mixture was stirred for 6 hours at 18°C. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a yellow solid. The crude was purified by silica gel chromatography (gradient: 0% to 5% ethyl acetate (in petroleum ether)) to give D5 (158g, 81% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d 6 )δ8.72(br s,1H),8.41(d,J=8.6Hz,1H),6.76(d,J=8.6Hz,1H),3.00(d,J=4.8Hz,3H).

[0708] Step 2: 6-Chloro-N 2 -Synthesis of methylpyridine-2,3-diamine (D6)

[0709] Iron powder (15.4 g, 276 mmol) was added to a mixture of D5 (15.8 g, 84.2 mmol) in acetic acid (100 mL). The reaction mixture was stirred at 80 ° C for 3 hours, then cooled to room temperature and filtered. The filter cake was washed with ethyl acetate (2×100). The combined organic layers were concentrated under reduced pressure and the crude was purified by silica gel chromatography (eluent: 1:1 ethyl acetate / petroleum ether) to give D6 (8.40 g, 63% yield) as a brown solid.1 H NMR (400 MHz, CHLOROFORM-d) δ 6.79 (d, J = 7.7 Hz, 1H), 6.49 (d, J = 7.7 Hz, 1H), 3.00 (s, 3H).

[0710] Step 3: Synthesis of 5-chloro-2-(chloromethyl)-3-methyl-3H-imidazo[4,5-b]pyridine (P24)

[0711] Chloroacetyl chloride (55.5 mL, 698 mmol) was added to a solution of D6 (50.0 g, 317 mmol) in 1,4-dioxane (1.2 L), and the reaction mixture was stirred at 15 ° C for 50 minutes. It was then concentrated under reduced pressure to give a brown solid, which was taken up in trifluoroacetic acid (1.2 L) and stirred at 80 ° C for 60 hours. The mixture was concentrated under reduced pressure to give a brown oil, which was diluted with ethyl acetate (1 L) and neutralized by adding saturated aqueous sodium bicarbonate solution. When the carbon dioxide evolution ceased, the layers were separated and the aqueous layer was extracted with ethyl acetate (200 ml). The organic extracts were combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by silica gel chromatography (gradient: 10% to 25% ethyl acetate (in petroleum ether)) to give P24 (61.0 g, 79% yield) as a yellow solid. LCMS m / z 215.7 (dichloro isotope pattern observed) [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ8.13(d,J=8.3Hz,1H),7.37(d,J=8.4Hz,1H),5.11(s,2H),3.84(s,3H).

[0712] Examples 1 and 2

[0713] 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperazin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, ENT-X1, trifluoroacetate (1) [from C39]; and 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperazin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, ENT-X2, trifluoroacetate (2) [from C40]

[0714]

[0715]

[0716] Step 1: Synthesis of methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperazin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (C38)

[0717] This experiment was performed in two batches of the same scale. A reaction vessel containing a mixture of C2 (500 mg, 1.52 mmol), P12 (530 mg, 1.59 mmol), [2',6'-bis(propan-2-yloxy)biphenyl-2-yl](dicyclohexyl)phosphane (Ruphos; 142 mg, 0.304 mmol), tris(dibenzylideneacetone)dipalladium(0) (139 mg, 0.152 mmol), and cesium carbonate (1.48 g, 4.54 mmol) in toluene (15 mL) was evacuated and filled with nitrogen. This evacuation cycle was repeated twice, whereupon the reaction mixture was stirred at 100° C. for 16 hours, combined with the second batch, and filtered. The filtrate was concentrated, and the residue was subjected to silica gel chromatography (gradient: 0% to 60% ethyl acetate in petroleum ether) and then preparative thin layer chromatography (eluent: 1:1 petroleum ether / ethyl acetate) to afford C38 as a light yellow solid. Combined yield: 600 mg, 1.03 mmol, 34%. LCMS m / z 581.0 ◆ [M+H] + .

[0718] Step 2: Separation of methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperazin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate, ENT-1 (C39) and methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperazin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate, ENT-2 (C40)

[0719] Separation of C38 (780 mg, 1.34 mmol) into its constituent enantiomers was performed using SFC [column: Chiral Technologies Chiralpak AD, 10 μm; mobile phase: 3:2 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting enantiomer was designated ENT-1 (C39), which was obtained as a white solid. Yield: 282 mg, 0.485 mmol, 36%. LCMS m / z 581.0◆[M+H] +Retention time 1.90 minutes (column: Chiral Technologies Chiralpak AD-3, 4.6 x 50 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: ethanol containing 0.05% diethylamine; gradient: 5% B in 0.20 minutes, then 5% to 40% B over 1.4 minutes, then maintained at 40% B for 1.05 minutes; flow rate: 4.0 mL / min).

[0720] The second eluting enantiomer was designated ENT-2 (C40), which was subjected to a second purification using SFC [column: Chiral Technologies Chiralpak AD, 10 μm; mobile phase: 3:2 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. This provided C40 as a light brown solid. Yield: 280 mg, 0.482 mmol, 36%. LCMS m / z 581.0 ◆ [M+H] + Retention time 2.18 minutes (analytical conditions were the same as those used for C39).

[0721] Step 3: Synthesis of 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperazin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, ENT-X1, trifluoroacetate (1) [from C39]

[0722] Aqueous lithium hydroxide solution (2M; 0.30 mL, 0.60 mmol) was added to a solution of C39 (70 mg, 0.12 mmol) in a mixture of methanol (3 mL) and tetrahydrofuran (3 mL). After the reaction mixture was stirred at 25 °C for 16 hours, aqueous lithium hydroxide solution (2M; 0.30 mL, 0.60 mmol) was added again, and stirring was continued for another 20 hours. The reaction mixture was then adjusted to pH 7 by adding 1 M hydrochloric acid, and then concentrated in vacuo to remove methanol and tetrahydrofuran. The residue was adjusted to pH 5-6 by adding trifluoroacetic acid, and then purified by reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.1% trifluoroacetic acid (in water); mobile phase B: acetonitrile; gradient: 30% to 60% B) to give 1 as a white solid. Yield: 40.5 mg, 59.5 μmol, 50%. LCMS m / z 567.0◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4)δ8.37(br s,1H),8.07(dd,J=8.5,1.5Hz,1H),7.79(d,J=8.6Hz,1H),7.59(dd,J=8.0,8.0Hz,1H),7.34(dd,J=10.2,2.0Hz,1H),7.30(br dd,J=8.3,2.0Hz,1H),7.22(s,1H),6.87(dd,J=8.1,8.1Hz,1H),6.63(br d,J=8Hz,1H),6.60(br d,J=8Hz,1H),4.70(s,2H),4.65(t,J=4.8Hz,2H),3.75(t,J=4.8Hz,2H),3.59-3.42(m,8H),3.29(s,3H).

[0723] Step 4: Synthesis of 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperazin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, ENT-X2, trifluoroacetate (2) [from C40]

[0724] Aqueous lithium hydroxide solution (2M; 0.30 mL, 0.60 mmol) was added to a solution of C40 (69 mg, 0.12 mmol) in a mixture of methanol (3 mL) and tetrahydrofuran (3 mL). After the reaction mixture was stirred at 25 °C for 16 hours, aqueous lithium hydroxide solution (2M; 0.30 mL, 0.60 mmol) was added again, and stirring was continued for another 20 hours. The reaction mixture was adjusted to pH 7 by adding 1 M hydrochloric acid, and then concentrated in vacuo to remove methanol and tetrahydrofuran. The residue was adjusted to pH 5-6 by adding trifluoroacetic acid, and then purified by reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.1% trifluoroacetic acid (in water); mobile phase B: acetonitrile; gradient: 30% to 60% B) to give 2 as a white solid. Yield: 22.9 mg, 33.6 μmol, 28%. LCMS m / z 567.0◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4)δ8.40-8.35(m,1H),8.07(dd,J=8.6,1.5Hz,1H),7.79(d,J=8.6Hz,1H),7.59(dd,J=8.0,8.0Hz,1H),7.35(dd,J=10.2,2.0Hz,1H),7.31(br dd,J=8,2Hz,1H),7.22(s,1H),6.87(dd,J=8.3,8.0Hz,1H),6.63(br d,J=8Hz,1H),6.60(br d,J=8Hz,1H),4.68(s,2H),4.65(t,J=4.9Hz,2H),3.76(t,J=4.8Hz,2H),3.57-3.40(m,8H),3.29(s,3H).

[0725] Example 3

[0726] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(2-methoxyethyl)-1H-imidazo[4,5-b]pyridine-6-carboxylic acid, trifluoroacetate (3)

[0727]

[0728] Step 1: Synthesis of 4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine (C13, free base)

[0729] p-Toluenesulfonic acid monohydrate (318 mg, 1.67 mmol) was added to a solution of P2 (300 mg, 0.670 mmol) in ethyl acetate (3.5 mL). The reaction mixture was stirred at 60 °C for 1 hour, whereupon it was basified by the addition of saturated aqueous potassium carbonate solution (20 mL) and extracted with a mixture of dichloromethane and methanol (10:1, 3 x 50 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo to provide C13, free base, as a brown solid. Yield: 230 mg, 0.661 mmol, 99%.

[0730] Step 2: Synthesis of 6-bromo-2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(2-methoxyethyl)-1H-imidazo[4,5-b]pyridine (C41)

[0731] A suspension of C13, free base (130 mg, 0.374 mmol), P13 (130 mg, 0.427 mmol), and potassium carbonate (172 mg, 1.24 mmol) in acetonitrile (2 mL) was stirred at 50 °C for 16 hours. The reaction mixture was then purified using preparative thin layer chromatography (eluent: ethyl acetate) to afford C41 as a brown oil. Yield: 114 mg, 0.185 mmol, 49%. LCMS m / z 617.1 (bromine-chlorine isotope pattern observed) [M+H] + .

[0732] Step 3: Synthesis of methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(2-methoxyethyl)-1H-imidazo[4,5-b]pyridine-6-carboxylate (C42)

[0733] A solution of C41 (114 mg, 0.185 mmol), 1,3-bis(diphenylphosphino)propane (15.3 mg, 37.1 μmol), palladium(II) acetate (8.3 mg, 37 μmol), and triethylamine (187 mg, 1.85 mmol) in a mixture of methanol (5 mL) and N,N-dimethylformamide (1 mL) was stirred at 80 °C under carbon monoxide (50 psi) for 16 hours. After the reaction mixture was diluted with ethyl acetate (50 mL), it was washed with saturated aqueous sodium chloride solution (2 x 50 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Purification was performed using preparative thin layer chromatography (eluent: ethyl acetate) to provide C42 as a colorless oil. Yield: 60.0 mg, 0.101 mmol, 55%. LCMS m / z 617.2 (chlorine isotope pattern observed) [M+Na + ].

[0734] Step 4: Synthesis of 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(2-methoxyethyl)-1H-imidazo[4,5-b]pyridine-6-carboxylic acid, trifluoroacetate (3)

[0735] Aqueous sodium hydroxide solution (3M; 1.0 mL, 3.0 mmol) was added to a solution of C42 (60.0 mg, 0.101 mmol) in methanol (2.0 mL), and the reaction mixture was stirred at 20 °C for 2 hours. It was then adjusted to pH 7 by the addition of 1 M hydrochloric acid and extracted with a mixture of dichloromethane and methanol (10:1, 3 x 30 mL). The combined organic layers were dried over magnesium sulfate, filtered, concentrated in vacuo, and purified using reverse phase HPLC (column: Boston Green ODS, 5 μm; mobile phase A: 0.1% trifluoroacetic acid in water; mobile phase B: acetonitrile; gradient: 10% to 95% B) to give 3 as a white solid. Yield: 29.6 mg, 42.6 μmol, 42%. LCMS m / z 581.0 ◆ [M+H] + . 1 H NMR (400 MHz, methanol-d 4 )δ9.13 (d, J = 1.9 Hz, 1H), 8.74 (d, J = 1.9 Hz, 1H), 7.63 (dd, J = 8.3, 8.3 Hz, 1H), 7.30 (dd, J = 10.9, 2.0 Hz, 1H), 7.24 (ddd, J = 8.4, 2.0, 0.7 Hz, 1H), 6.89-6.84 (m, 1H), 6.82-6.77 (m, 2H), 4.98-4.89 (m, 2H, assumed; mostly obscured by water peak), 4.64 (t, J = 4.8 Hz, 2H), 4.04-3.92 (br m,2H),3.75(dd,J=5.4,4.2Hz,2H),3.51-3.39(m,2H),3.31(s,3H),3.19- 3.06(m,1H),2.41-2.24(m,2H),2.24-2.12(m,2H),2.06(d,J=1.0Hz,3H).

[0736] Examples 4 and 5

[0737] 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (4) and 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (5)

[0738]

[0739] Step 1: Synthesis of 4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidine, trifluoroacetate (C43)

[0740] Trifluoroacetic acid (1.3 mL) was added to a solution of P1 (300 mg, 0.691 mmol) in dichloromethane (5 mL). The reaction mixture was stirred at 29 °C for 2 h, whereupon it was concentrated in vacuo to give C43 as a brown oil, which was used directly in the next step.

[0741] Step 2: Synthesis of methyl 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C44) and methyl 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C45)

[0742] P15 (204 mg, 0.692 mmol) was added to a solution of C43 (from the previous step, ≤ 0.691 mmol) in acetonitrile (10 mL), followed by potassium carbonate (956 mg, 6.92 mmol). The reaction mixture was stirred at 29 °C for 16 hours, whereupon it was filtered; the filtrate was concentrated in vacuo to give a residue which was purified by preparative thin layer chromatography (eluent: 2:1 petroleum ether / ethyl acetate) to provide a mixture of diastereomeric products as a yellow gum (178 mg). The two products were separated via SFC [column: Chiral Technologies ChiralCel OD, 5 μm; mobile phase: 55:45 carbon dioxide / (methanol containing 0.1% ammonium hydroxide)]. The first eluting diastereomer obtained as a yellow oil was designated C44. Yield: 44.3 mg, 74.8 μmol, 11% (over two steps). LCMS m / z 592.1◆[M+H] + Retention time 4.26 minutes (column: Chiral Technologies ChiralCel OD-3, 4.6 x 100 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: methanol containing 0.05% diethylamine; gradient: 5% to 40% B over 4.5 minutes, then maintain at 40% B for 2.5 minutes; flow rate: 2.8 mL / min).

[0743] The second eluting diastereomer was subjected to a second purification via SFC [column: Chiral Technologies ChiralCel OD, 5 μm; mobile phase: 3:2 carbon dioxide / (methanol containing 0.1% ammonium hydroxide)] to provide the second eluting diastereomer as a colorless oil, which was designated C45. Yield: 38 mg, 64 μmol, 9% (over two steps). LCMS m / z 592.1◆[M+H] + Retention time 4.41 minutes (analytical conditions were the same as those used for C44).

[0744] The absolute stereochemistry expressed at the dioxolane was assigned via potency correlation of 5 with 5, a free acid sample (synthesized from intermediate C48); the absolute stereochemistry of the intermediate was determined via single crystal X-ray structure determination of C49, the hemisulfate salt of C48 (see below).

[0745] Step 3: Synthesis of 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (4)

[0746] Aqueous lithium hydroxide (2M; 0.80 mL, 1.6 mmol) was added to a solution of C44 (44.3 mg, 74.8 μmol) in a mixture of methanol (1 mL) and tetrahydrofuran (1 mL), and the reaction mixture was stirred at 26° C. for 3 hours. It was then adjusted to pH 7 by the addition of trifluoroacetic acid, and the resulting mixture was concentrated in vacuo and purified using reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B: acetonitrile; gradient: 30% to 50% B) to give 4 as a white solid. Yield: 26.6 mg, 44.7 μmol, 60%. LCMS m / z 578.0◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4)δ8.31(d,J=1.4Hz,1H),7.96(dd,J=8.5,1.6Hz,1H),7.66(d,J=8.5Hz,1H),7.57(dd,J=8.0,8.0Hz,1H),7.34(dd,J=10.1,2.0Hz,1H),7.29(br dd,J=8.3,2.0Hz,1H),7.20(s,1H),6.86-6.79(m,1H),6.77(br dd, component of ABC mode, J = 7.9, 1.3 Hz, 1H), 6.73 (dd, component of ABC mode, J = 7.5, 1.4 Hz, 1H), 5.29-5.18 (m, 1H), 4.9-4.78 (m, 1H, assumed; partially obscured by water peak), 4.68 (dd, J = 15.3, 2.7 Hz, 1H), 4.54 (td, J = 8.0, 5.9 Hz, 1H), 4.44 (dt, J = 9.2, 5.9 Hz, 1H), 4.02 (AB quartet, J AB =13.9Hz,Δν AB =49.0Hz,2H),3.18-3.08(m,1H),3.05-2.96(m,1H),2.81-2.68(m,2H),2. 56-2.45(m,1H),2.45-2.30(m,2H),2.03-1.88(m,2H),1.88-1.79(m,2H).

[0747] Step 4: Synthesis of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (5)

[0748] Aqueous lithium hydroxide solution (2M; 0.80 mL, 1.6 mmol) was added to a solution of C45 (38 mg, 64 μmol) in a mixture of methanol (1 mL) and tetrahydrofuran (1 mL), and the reaction mixture was stirred for 2.5 hours at 24°C. It was then adjusted to pH 7 by the addition of 1 M hydrochloric acid, and the resulting mixture was concentrated in vacuo and purified using reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.05 ammonium hydroxide (in water); mobile phase B: acetonitrile; gradient: 29% to 49% B) to provide 5 as a white solid. Yield: 27.9 mg, 46.9 μmol, 73%. LCMS m / z 577.9◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4)δ8.32(d,J=1.4Hz,1H),7.96(dd,J=8.5,1.5Hz,1H),7.66(d,J=8.5Hz,1H),7.56(dd,J=8.0,8.0Hz,1H),7.34(dd,J=10.2,2.0Hz,1H),7.29(br dd, J = 8.3, 2.0 Hz, 1H), 7.20 (s, 1H), 6.85-6.80 (m, 1H), 6.77 (dd, component of ABC mode, J = 8.0, 1.3 Hz, 1H), 6.73 (dd, component of ABC mode, J = 7.5, 1.4 Hz, 1H), 5.30-5.20 (m, 1H), 4.9-4.79 (m, 1H, assumed; partially obscured by water peak), 4.68 (dd, J = 15.4, 2.7 Hz, 1H), 4.62-4.54 (m, 1H), 4.44 (dt, J = 9.2, 5.9 Hz, 1H), 4.02 (AB quartet, J AB =13.9Hz,Δν AB =44.6Hz,2H),3.18-3.09(m,1H),3.06-2.97(m,1H),2.80-2.67(m,2H),2.55-2.30(m,3H),2.02-1.78(m,4H).

[0749] Example 5, Alternative Synthesis of Free Acid

[0750] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (5, free acid)

[0751]

[0752] Step 1: Separation of tert-butyl 4-[(2R)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (C46) and tert-butyl 4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidine-1-carboxylate (C47)

[0753] Separation of P1 (10 g, 23 mmol) into its constituent enantiomers was performed using reverse phase HPLC [column: Phenomenex Lux Amylose-1, 5 μm; mobile phase: 9:1 carbon dioxide / (2-propanol containing 0.2% 1-aminopropan-2-ol)]. The first eluting enantiomer was designated as C46 and the second eluting enantiomer as C47, both obtained as colorless oils. The absolute stereochemistry indicated for C46 and C47 was assigned based on single crystal X-ray structure determination performed on C49 (synthesized from C47) (see below).

[0754] C46 yield: 4.47 g, 10.3 mmol, 45%. Retention time: 3.98 min [column: Phenomenex LuxAmylose-1, 4.6 x 250 mm, 5 μm; mobile phase A: carbon dioxide; mobile phase B: 2-propanol containing 0.2% 1-aminopropan-2-ol; gradient: 5% B, 1.00 min, then 5% to 60% B, over 8.00 min; flow rate: 3.0 mL / min; back pressure: 120 bar].

[0755] C47 yield: 4.49 g, 10.3 mmol, 45%. Retention time: 4.32 min (Analytical SFC conditions were identical to those used for C46).

[0756] Step 2: Synthesis of 4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidine (C48)

[0757] p-Toluenesulfonic acid monohydrate (566 mg, 2.98 mmol) was added to a solution of C47 (1.12 g, 2.58 mmol) in ethyl acetate (26 mL). After the reaction mixture was heated at 45 °C for 16 hours, it was concentrated in vacuo, dissolved in ethyl acetate, and washed with saturated aqueous sodium bicarbonate solution. The aqueous layer was extracted with ethyl acetate, and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered and concentrated under reduced pressure to give C48 (947 mg) as a foamy white solid, LCMS m / z 334.0 [M+H] + A portion of this material, which still contained some p-toluenesulfonic acid, was used in the synthesis of C50 below.

[0758] The second portion of the foamy white solid (440 mg) was dissolved in ethyl acetate (25 mL) and washed with saturated aqueous sodium bicarbonate solution (2 x 15 mL); the organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo to give C48 (350 mg) as a colorless oil that no longer contained p-toluenesulfonic acid. Adjusted yield: 350 mg, 1.05 mmol, 88%. 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.53 (dd, J = 8.4, 7.8 Hz, 1H), 7.22-7.13 (m, 3H), 6.87-6.80 (m, 1H), 6.79-6.71 (m, 2H), 3.23-3.14 (m, 2H), 2.86-2.69 (m, 3H), 1.90-1.68 (m, 4H).

[0759] Step 3: Synthesis of 4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidine, hemisulfate salt (C49)

[0760] A 0.1 M solution of C48 (colorless oil from before) in ethyl acetate was prepared and subjected to salt screening. Only sulfate formation is described here. A mixture of sulfuric acid (25 μmol) and substrate solution (0.1 M, 250 μL, 25 μmol) was heated to 45°C for 1 hour, allowed to cool to room temperature, and stirred for 15 hours. The resulting suspension was treated with methanol (approximately 150 μL) until a solution was formed; it was allowed to evaporate slowly overnight until approximately 50 μL of solvent remained. One of the resulting crystals was analyzed by single crystal X-ray structure determination, and the absolute stereochemistry was established as that shown.

[0761] Single crystal X-ray structure determination of C49

[0762] Single crystal X-ray analysis

[0763] Data collection was performed at room temperature on a Bruker D8 Venture diffractometer. Data collection consisted of ω and φ scans.

[0764] Through the intrinsic phase, the SHELX software was used to fit the triclinic space group P 1 The structure was solved in . Then, the structure was refined by full-matrix least squares. All non-hydrogen atoms were found and refined using anisotropic displacement parameters.

[0765] The hydrogen atoms located on the nitrogen were found from the Fourier difference map and refined with restrained distances. The remaining hydrogen atoms were placed at the calculated positions and carried on their supporting atoms. The final refinement included isotropic displacement parameters for all hydrogen atoms.

[0766] The asymmetric unit is composed of two molecules of protonated C48, one molecule of doubly deprotonated sulfuric acid, and one molecule of fully occupied water. Thus, the structure is a hemisulfate and a hemihydrate. The chlorofluorophenyl ring is disordered and modeled with a 60 / 40 occupancy, with the ring flipped in two positions.

[0767] Absolute structure analysis using the likelihood method (Hooft, 2008) was performed using PLATON (Spek). The results indicated that the absolute structure had been correctly assigned; the method calculated the probability that the structure was correct to be 100.0. The Hooft parameter was reported as 0.061 with an esd of 0.004, and the Parson parameter was reported as 0.063 with an esd of 0.005.

[0768] The final R index was 3.1%. The final difference Fourier analysis showed no missing or misplaced electron density.

[0769] Relevant crystal, data collection, and refinement information are summarized in Table E. Atomic coordinates, bond lengths, bond angles, and displacement parameters are listed in Tables FH.

[0770] Software and References

[0771] SHELXTL, Version 5.1, Bruker AXS, 1997.

[0772] PLATON,ALSpek,J.Appl.Cryst.2003,36,7-13.

[0773] MERCURY, CFMacrae, PREdington, P.McCabe, E.Pidcock, GPShields, R.Taylor, M.Towler and J.van de Streek, J.Appl.Cryst.2006, 39, 453-457.

[0774] OLEX2, OV Dolomanov, LJ Bourhis, RJ Gildea, JAK Howard and H. Puschmann, J. Appl. Cryst. 2009, 42, 339-341.

[0775] RWWHooft, LHStraver and ALSpek, J. Appl. Cryst. 2008, 41, 96-103.

[0776] HDFlack, Acta Cryst. 1983, A39, 867-881.

[0777] Table E: Crystal data and structure refinement of C49.

[0778]

[0779] Table F: Atomic coordinates of C49 (x 10 4 ) and the equivalent isotropic displacement parameter U(eq) is defined as the orthogonal U ij One third of the tensor's trace.

[0780]

[0781]

[0782]

[0783] Table G: Bond lengths of C49 and angle [°].

[0784]

[0785]

[0786]

[0787]

[0788]

[0789]

[0790]

[0791]

[0792]

[0793]

[0794] Symmetry transformations used to produce equivalent atoms.

[0795] Table H: Anisotropic displacement parameters of C49 The anisotropic displacement factor exponent takes the following form: -2π 2 [h 2 a* 2 U 11 +...+2h ka*b*U 12 ].

[0796]

[0797]

[0798]

[0799] Step 4: Synthesis of methyl 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C50)

[0800] A solution of C48 (500 mg, from the previous foamy white solid, corrected for p-toluenesulfonic acid: 1.25 mmol) in acetonitrile (6 mL) was treated with N,N-diisopropylethylamine (0.68 mL, 3.9 mmol) and allowed to stir at 45 ° C for 5 minutes. After adding P15 (319 mg, 1.08 mmol), stirring was continued at 45 ° C for 7.25 hours, whereupon the reaction mixture was diluted with water (6 mL) and acetonitrile (2 mL) at 45 ° C. The resulting heterogeneous mixture was cooled to room temperature and stirred for 72 hours. More water (5 mL) was added, and after a further 30 minutes of stirring, the solid was collected via filtration and washed with a mixture of acetonitrile and water (15:85, 3x 5 mL) to give C50 as a white solid with a slight pink character. Yield: 605 mg, 1.02 mmol, 82%. LCMS m / z 592.0◆[M+H] + . 1 H NMR (400 MHz, chloroform-d) δ 8.17 (d, J = 1.6 Hz, 1H), 7.96 (dd, J = 8.5, 1.5 Hz, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.51 (dd, J = 8.0, 8.0 Hz, 1H), 7.19 (br s, 1H), 7.18-7.14 (m, 2H), 6.85-6.79 (m, 1H), 6.76-6.71 (m, 2H), 5.26-5.18 (m, 1H), 4.73 (dd, ABX pattern component, J = 15.3, 5.9 Hz, 1H), 4.67 (dd, ABX pattern component, J = 15.3, 3.5 Hz, 1H) ,4.63-4.55(m,1H),4.38(ddd,J=9.1,6.0,5.9Hz,1H),3.94(s,5H),3.03-2.89(m ,2H),2.77-2.65(m,2H),2.51-2.39(m,1H),2.34-2.20(m,2H),1.91-1.76(m,4H).

[0801] Step 5: Synthesis of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (5, free acid)

[0802] A suspension of C50 (595 mg, 1.00 mmol) in methanol (10 mL) was heated to 45 °C and treated with aqueous sodium hydroxide (1 M; 2.01 mL, 2.01 mmol). After 21 hours at 45 °C, the reaction mixture was cooled to room temperature; it was then treated with aqueous citric acid (1 M, 1 mL) to a pH of 5 to 6. Water (10 mL) was added and the mixture was stirred for 1 hour, whereupon the solid was collected by filtration. It was washed with a mixture of methanol and water (1:10, 3 x 5 mL) to give a solid (433 mg). A portion of this material (300 mg) was stirred with a mixture of heptane and ethyl acetate (1:3, 5 mL) at 40 °C for 1 hour; after cooling to room temperature with continued stirring, the solid was collected by filtration and washed with a mixture of heptane and ethyl acetate (3:1, 3 x 3 mL) to give 5' free acid as a white solid. Yield: 260 mg, 0.450 mmol, corresponding to 65% for the entire reaction. LCMS m / z 578.0◆[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ12.75(v br s,1H),8.26(br s,1H),7.79(dd,J=8.4,1.6Hz,1H),7.66-7.56(m,3H),7.40(dd,J=8.3,2.0Hz,1H),7.35(s,1H),6.87-6.75(m,3H),5.13-5.03(m,1H),4.76(dd,component of ABX mode,J=15.3,7.2Hz,1H),4.62(dd,component of ABX mode,J=15.2,2.8Hz,1H),4.46-4.38(m,1H),4.34(ddd,J=9.0,5.9,5.8Hz,1H),3.84(AB quartet,J AB =13.5Hz,Δν AB =67.7Hz,2H),3.00(br d,J=11.2Hz,1H),2.84br(d,J=11.3Hz,1H),2.71-2.56(m,2H),2.45-2.34(m,1H),2.28-2.08(m,2H),1.84-1.65(m,4H).

[0803] This material was determined to have the same absolute configuration as Example 5 above by comparison of its biological activity with that of 4 and 5. In Assay 2, a sample of this 5, the free acid, exhibited an EC of 25 nM. 50 (Geometric mean of 3 replicates). The activities of the ammonium salts of Example 4 and Example 5 in Assay 2 were >20000 nM (2 replicates) and 20 nM (geometric mean of 3 replicates), respectively.

[0804] Example 5, Synthesis of 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine salt

[0805] 1,3-Dihydroxy-2-(hydroxymethyl)propan-2-amine 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (5,1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine)

[0806]

[0807] A mixture of 5, free acid (0.50 g, 0.86 mmol) in tetrahydrofuran (4 mL) was treated with an aqueous solution of 2-amino-2-(hydroxymethyl)propane-1,3-diol (Tris, 1.0 M; 0.5 mL, 1.0 mmol). After 20 hours, the mixture was concentrated in vacuo with ethanol (2 x 6 mL). The mixture was treated with ethanol (4 mL). After stirring for 48 hours, the solid was collected by filtration, washed with ethanol (2 x 10 mL) and dried in vacuo to obtain 5,1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine salt as a white solid. Yield: 410 mg, 0.586 mmol, 68%. 1 H NMR (600 MHz, DMSO-d 6), characteristic peaks: δ8.19 (s, 1H), 7.78 (br d, J = 8.4 Hz, 1H), 7.62-7.58 (m, 2H), 7.55 (d, J = 8.3 Hz, 1H), 7.40 (dd, J = 8.4, 2.0 Hz, 1H), 7.35 (s, 1H), 6.85-6.80 (m, 2H), 6.79 (dd, J = 6.9, 2.4 Hz, 1H), 5.11-5.05 (m, 1H), 4 .73(dd,J=15.2,7.2Hz,1H),4.60(dd,J=15.3,2.9Hz,1H),4.45-4.39(m,1H),4.34( ddd,J=9.0,6.0,5.8Hz,1H),3.91(d,J=13.5Hz,1H),3.74(d,J=13.5Hz,1H),2.99(br d,J=11.1Hz,1H),2.85(br d,J=11.3Hz,1H),2.68-2.59(m,2H),2.44-2.37(m,1H),2.25-2.18(m,1H),2.17-2.10(m,1H),1.80-1.69(m,4H). mp = 168°C to 178°C.

[0808] Examples 6 and 7

[0809] 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (6) and 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (7)

[0810]

[0811] Step 1: Synthesis of 4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidine, p-toluenesulfonate (C13)

[0812] A solution of P2 (150 mg, 0.335 mmol) and p-toluenesulfonic acid monohydrate (159 mg, 0.836 mmol) in ethyl acetate (2.0 mL) was stirred at 60° C. for 3.5 hours. The reaction mixture was concentrated in vacuo to give C13 as a brown oil, which was used directly in the next step. LCMS m / z 348.1◆[M+H] + .

[0813] Step 2: Synthesis of methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C51)

[0814] P15 (99.1 mg, 0.336 mmol) was added to a suspension of C13 (from the previous step; ≤ 0.335 mmol) and potassium carbonate (232 mg, 1.68 mmol) in acetonitrile (5.0 mL). The reaction mixture was stirred at 60 °C for 10 hours, whereupon it was filtered and the filtrate concentrated in vacuo. After the residue (390 mg) was combined with material from a similar reaction performed using C13 (≤ 0.11 mmol), it was diluted with water (20 mL) and extracted with a mixture of dichloromethane and methanol (10: 1, 3 x 30 mL). The combined organic layers were dried over sodium sulfate, filtered, concentrated in vacuo, and subjected to preparative thin layer chromatography (eluent: 1: 1 dichloromethane / methanol) to provide C51, a mixture of diastereomers, as a colorless oil. Combined yield: 80.6 mg, 0.133 mmol, 30% (over two steps). LCMS m / z 606.2◆[M+H] + .

[0815] Step 3: Separation of methyl 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C52) and methyl 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C53)

[0816] C51 (180 mg, 0.297 mmol) was separated into its constituent diastereomers via repeated SFC [column: Chiral Technologies Chiralpak AD, 10 μm; mobile phase: 65:35 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting diastereomer was designated C52. Yield: 61.2 mg, 0.101 mmol, 34%. LCMS m / z 627.9 ◆ [M+Na +]. Retention time: 5.03 minutes (column: Chiral Technologies Chiralpak AD-3, 4.6 x 150 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: ethanol containing 0.05% diethylamine; gradient: 5% to 40% B over 5.5 minutes, then maintained at 40% B for 3.0 minutes; flow rate: 2.5 mL / min).

[0817] The second eluting diastereomer was designated C53. Upon analysis, this material was confirmed to be contaminated with the corresponding ethyl ester; it was taken to the hydrolysis step (to produce 7) as a mixture. Yield: 40.0 mg, 66.0 μmol, 22%. LCMS m / z 606.0 ◆ [M+H] + Retention time: 5.19 minutes (analytical conditions were the same as those for C52).

[0818] The absolute stereochemistry expressed at the dioxolane was assigned via potency correlation of 7 with a sample of 7, the free acid (synthesized from intermediate P3, see Example 7, alternative synthesis of the free acid, below); the absolute stereochemistry of P3 was determined via single crystal X-ray structure determination of C8 (see below).

[0819] Step 4: Synthesis of 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (6)

[0820] Aqueous lithium hydroxide (2M; 0.990 mL, 1.98 mmol) was added to a solution of C52 (60 mg, 99 μmol) in a mixture of methanol (1.0 mL) and tetrahydrofuran (1.0 mL), and the reaction mixture was stirred at 20° C. for 16 hours. Trifluoroacetic acid was added until the pH of the reaction mixture reached 7, whereupon it was concentrated in vacuo, and the residue was purified using reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B: acetonitrile; gradient: 29% to 49% B) to give 6 as a white solid. Yield: 14.4 mg, 23.6 μmol, 24%. LCMS m / z 592.0◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4), characteristic peaks: δ8.35 (d, J = 1.3 Hz, 1H), 7.97 (dd, J = 8.5, 1.5 Hz, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.58 (dd, J = 8.3, 8.3 Hz, 1H), 7.28 (dd, J = 10.9, 2.0 Hz, 1H), 7.21 (br dd, J = 8.4, 1.9 Hz, 1H), 6.81-6.75 (m, 1H), 6.74-6.68 (m, 2H), 5.33-5.25 (m, 1H), 4.72 (dd, J = 15.4, 2.7 Hz, 1H), 4.49 (dt, J = 9.1, 6.0 Hz, 1H), 4.03 (AB quartet, J AB =13.9Hz,Δν AB =47.8Hz,2H),3.14(br d,J=11Hz,1H),3.02(br d,J=11.5Hz,1H),2.88-2.78(m,1H),2.77-2.68(m,1H),2.60-2.50(m,1H), 2.47-2.32(m,2H),2.03(d,J=1.1Hz,3H),2.01-1.87(m,2H),1.87-1.78(br m,2H).

[0821] Step 5: Synthesis of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid ammonium (7)

[0822] Aqueous lithium hydroxide solution (2M; 0.642 mL, 1.28 mmol) was added to a solution of C53 (38.9 mg, 64.2 μmol) in a mixture of methanol (1.0 mL) and tetrahydrofuran (1.0 mL). After the reaction mixture was stirred at 20 °C for 16 hours, it was adjusted to pH 7 by the addition of trifluoroacetic acid, concentrated in vacuo, and purified using reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B: acetonitrile; gradient: 0% to 80% B) to give 7 as a white solid. Yield: 25.1 mg, 41.2 μmol, 64%. LCMS m / z 591.9 ◆ [M+H] + . 1 H NMR (400 MHz, methanol-d 4), characteristic peaks: δ8.34 (d, J = 1.5 Hz, 1H), 7.98 (dd, J = 8.5, 1.6 Hz, 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.58 (dd, J = 8.3, 8.3 Hz, 1H), 7.28 (dd, J = 10.9, 2.0 Hz, 1H), 7.20 (br dd, J = 8.4, 1.9 Hz, 1H), 6.81-6.74 (m, 1H), 6.74-6.67 (m, 2H), 5.33-5.23 (m, 1H), 4.73 (dd, J = 15.4, 2.7 Hz, 1H), 4.68-4.61 (m, 1H), 4.48 (dt, J = 9.1, 5.9 Hz, 1H), 4.05 (AB quartet, J AB =13.9Hz,Δν AB =44.1Hz,2H),3.15(br d,J=11.7Hz,1H),3.03(br d,J=11.6Hz,1H),2.87-2.69(m,2H),2.60-2.49(m,1H),2.48-2.33(m,2H),2.03(br s,3H),2.01-1.77(m,4H).

[0823] Example 7, Alternative Synthesis of Free Acid

[0824] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (7, free acid)

[0825]

[0826] Step 1: Synthesis of methyl 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C53)

[0827] N,N-diisopropylethylamine (15.1 mL, 86.9 mmol) was added to a mixture of P3 (8.22 g, 15.8 mmol) in acetonitrile (185 mL); after stirring for 5 minutes, P15 (4.57 g, 15.5 mmol) was added and the reaction mixture was heated at 45 °C. After 4 hours, the reaction mixture was concentrated to half of its original volume in vacuo, and the resulting mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 100 mL). The combined organic layers were washed with water (50 mL), dried over magnesium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (gradient: 50% to 100% ethyl acetate (in heptane)) gave C53 as a white solid. Yield: 8.4 g, 13.9 mmol, 88%. LCMS m / z 606.1 ◆ [M+H] + . 1 H NMR (600 MHz, DMSO-d 6 )δ8.30(s,1H),7.82(br d,J=8.4Hz,1H),7.67(d,J=8.4Hz,1H),7.58-7.53(m,2H),7.33(dd,J=8.4,2.1Hz ,1H),6.80-6.76(m,2H),6.76-6.72(m,1H),5.14-5.07(m,1H),4.81(dd,J=15.2,7 .2Hz,1H),4.67(dd,J=15.3,2.8Hz,1H),4.51-4.44(m,1H),4.37(ddd,J=8.9,5.9 ,5.9Hz,1H),3.97(d,J=13.6Hz,1H),3.87(s,3H),3.78(d,J=13.5Hz,1H),3.02(br d,J=11.1Hz,1H),2.86(br d,J=11.3Hz,1H),2.74-2.60(m,2H),2.48-2.41(m,1H),2.29-2.22(m,1H ),2.21-2.14(m,1H),2.02(s,3H),1.83-1.73(m,2H),1.73-1.64(m,2H).

[0828] Step 2: Synthesis of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (7, free acid)

[0829] A mixture of C53 (8.40 g, 14.0 mmol) in methanol (135 mL) was heated to 45 °C and treated with aqueous sodium hydroxide solution (1 M; 27.7 mL, 27.7 mmol). After 20 hours, the reaction mixture was concentrated to half of its original volume in vacuo. The resulting mixture was diluted with water (100 mL) and the pH was adjusted to 5-6 using aqueous citric acid solution (1 M, 15 mL). The resulting solid was filtered, washed with water (2 x 15 mL), and transferred to a separatory funnel as a solution in ethyl acetate (50 mL); in this way, residual water was removed. The organic layer was dried over magnesium sulfate, filtered, and combined with four batches prepared previously by a similar procedure (the amount of C53 used for these reactions was 987 mg, 1.63 mmol; 1.15 g, 1.90 mmol; 8.57 g, 14.1 mmol; and 12.6 g, 20.8 mmol) and concentrated in vacuo. The resulting sticky solid was treated with 10% ethyl acetate in heptane (500 mL). After 4 hours, the solid was collected by filtration and washed with 10% ethyl acetate in heptane (2 x 25 mL) to give 7, the free acid, as a white solid. Yield: 29.4 g, 0.527 mmol, 74% (for combined reactions). LCMS 592.2 ◆ [M+H] + . 1 H NMR (600 MHz, DMSO-d 6): δ12.74(br s,1H),8.28(s,1H),7.80(br d,J=8.4Hz,1H),7.64(d,J=8.4Hz,1H),7.59-7.52(m,2H),7.33(dd,J=8.4,2.1Hz,1H),6.81-6.76(m,2H),6.76-6.72(m,1H),5.14-5 .07(m,1H),4.79(dd,J=15.3,7.3Hz,1H),4.65(dd,J=15.2,2.8Hz,1H),4.51-4.45(m,1H),4.38(ddd,J=9.0,5.9,5.9Hz,1H),3.96(br d, J = 13.6 Hz, 1H), 3.78 (br d, J = 13.5 Hz, 1H), 3.02 (br d, J = 11.1 Hz, 1H), 2.86 (br d, J = 11.1 Hz, 1H), 2.74-2.60 (m, 2H), 2.48-2.41 (m, 1H), 2.29-2.21 (m, 1H), 2.21-2.14 (m, 1H), 2.02 (s, 3H), 1.83-1.74 (m, 2H), 1.74-1.64 (m, 2H). By comparison of its biological activity with that of 6 and 7, this substance was determined to have the same absolute configuration as Example 7 above. In Assay 2, this 7, a sample of the free acid, exhibited an EC of 4.3 nM. 50 (Geometric mean of 3 replicates). The activities of the ammonium salts of Example 6 and Example 7 in Assay 2 were 2400 nM (geometric mean of 5 replicates) and 2.9 nM (geometric mean of 8 replicates), respectively.

[0830] Synthesis 7S-1. Example 7, Synthesis of 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine salt

[0831] 1,3-Dihydroxy-2-(hydroxymethyl)propan-2-amine 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (7,1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt)

[0832]

[0833] A mixture of 7, free acid (2.00 g, 3.38 mmol) in tetrahydrofuran (16 mL) was treated with an aqueous solution of 2-amino-2-(hydroxymethyl)propane-1,3-diol (Tris, 1.0 M; 3.55 mL, 3.55 mmol). After 18 hours, the reaction mixture was concentrated in vacuo and treated with ethanol (30 mL). After stirring this mixture for 23 hours, the solid was collected by filtration and washed with ethyl acetate (2 x 10 mL) to obtain 7, 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine salt as a white solid. Yield: 1.41 g, 1.98 mmol, 59%. LCMS m / z 592.3 ◆ [M+H] + . 1 H NMR (600 MHz, DMSO-d 6 ), characteristic peaks: δ8.20 (s, 1H), 7.79 (d, J = 8.4 Hz, 1H), 7.59-7.52 (m, 3H), 7.33 (br d, J = 8.5 Hz, 1H), 6.81-6.72 (m, 3H), 5.14-5.07 (m, 1H), 4.76 (dd, J = 15.2, 7.2 Hz, 1H), 4.63 (br d, J = 15.4 Hz, 1H), 4.50-4.44 (m, 1H), 4.37 (ddd, J = 8.9, 5.9, 5.9 Hz, 1H), 3.94 (d, J = 13.4 Hz, 1H), 3.76 (d, J = 13.4 Hz, 1H), 3.01 (br d, J = 11.1 Hz, 1H), 2.86 (br d,J=11.2Hz,1H),2.73-2.60(m,2H),2.5-2.41(m,1H),2.27-2.20(m,1H),2.20-2.13(m,1H),2.02(s,3H),1.83-1.64(m,4H). mp=175℃ to 180℃.

[0834] Synthesis of 7S-2: Example 7, Alternative Synthesis of 1,3-Dihydroxy-2-(Hydroxymethyl)propan-2-amine Salt

[0835] A 3.3M solution of 2-amino-2-(hydroxymethyl)-1,3-propanediol (1.0 equiv., 1.93 L) in water was added to a solution of 7, free acid (3.74 kg) in isopropanol (20 L) at 65 °C. Additional isopropanol (19 L) was added, followed by methanol (19 L) while maintaining the temperature at 65 °C. Over 2 hours, the mixture was slowly cooled to 45 °C and then maintained at 45 °C for at least 12 hours. Then, over 3 hours, the mixture was cooled to 5 °C and then maintained at 5 °C for at least 3 hours. The mixture was then filtered and the solids were collected and washed with ethyl acetate (2 x 10 mL) to give 7,1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt as a white solid (yield: 3.64 kg, 80.9%). LCMS and 1 H NMR data, which are essentially the same as those shown above for the synthesis of 7S-1.

[0836] Example 7, Obtaining Powder X-ray Diffraction (PXRD) Data of Form I of 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine Salt

[0837] The white solid of the tris salt of Example 7 (from Synthesis 7S-1 and Synthesis 7S-2) was subjected to PXRD analysis and found to be a crystalline material (designated as Form I of this anhydrous crystalline form). Powder X-ray diffraction analysis was performed using a Bruker AXSD8 Endeavor diffractometer equipped with a Cu radiation source. The divergence slit was set at 15 mm for continuous illumination. The diffracted radiation was detected by a PSD-Lynx Eye detector, with the detector PSD open set at 2.99 degrees. The X-ray tube voltage and current intensity were set at 40 kV and 40 mA, respectively. At the Cu wavelength (CuK ā=1.5418λ), in the θ-θ goniometer, using a 0.01 degree step size and a 1.0 second step time, from 3.0 to 40.0 degrees 2θ data were collected. The anti-scatter screen was set to a fixed distance of 1.5 mm. During data collection, the sample was rotated. The sample was prepared by placing it in a silicon low background sample holder and rotating it during collection. Using Bruker DIFFRAC Plus software, data were collected and analyzed by EVADiffrac Plus software. Before the peak search, the PXRD data file was not processed. Using the peak search algorithm in the EVA software, the peak selected by threshold 1 was used for preliminary peak assignment. To ensure validity, adjustments were made manually; the output of the automatic assignment was visually inspected, and the peak position was adjusted to the peak maximum. Peaks with relative intensities ≥3% are usually selected. In general, unresolved peaks or peaks consistent with noise are not selected. The typical error associated with the peak position of PXRD as specified by USP is up to + / -0.2° 2-θ (USP-941). One diffraction pattern was always observed and provided in Figure 1 The PXRD diffraction peaks expressed in degrees 2θ and the relative intensities (relative intensity ≥ 3%) of the samples synthesized above for 7S-2 are listed in Table X1.

[0838] Table X1

[0839]

[0840]

[0841] Examples 8 and 9

[0842] 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X1(8) [from C56]; and 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2(9) [from C57]

[0843]

[0844] Step 1: Synthesis of 3-fluoro-4-[2-methyl-4-(piperidin-4-yl)-1,3-benzodioxol-2-yl]benzonitrile, p-toluenesulfonate (C54)

[0845] p-Toluenesulfonic acid (158 mg, 0.919 mmol) was added to a solution of P4 (161 mg, 0.367 mmol) in ethyl acetate (8 mL), and the reaction mixture was stirred at 65 °C for 16 h. The solvent was removed in vacuo to provide C54 as a dark yellow gum; this material was used directly in the next step.

[0846] Step 2: Synthesis of methyl 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate (C55)

[0847] Potassium carbonate (219 mg, 1.58 mmol) was added to a solution of C54 (from the previous step; ≤ 0.367 mmol) in acetonitrile (3.7 mL), followed by P15 (115 mg, 0.390 mmol). The reaction mixture was stirred at 50 °C for 20 hours, whereupon it was diluted with ethyl acetate (10 mL) and filtered. The filter cake was washed with ethyl acetate (3 x 10 mL), and the combined filtrate was concentrated in vacuo. Silica gel chromatography (gradient: 0% to 100% ethyl acetate in petroleum ether) afforded C55 as a dark yellow oil. Yield: 191.0 mg, 0.320 mmol, 87% (over two steps). LCMS m / z 619.1 [M+Na + ].

[0848] Step 3: Separation of methyl 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate, ENT-1 (C56) and methyl 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate, ENT-2 (C57)

[0849] C55 (191 mg, 0.320 mmol) was separated into its constituent stereoisomers in dioxolane via SFC [column: Chiral Technologies ChiralCel OD, 5 μm; mobile phase: 3:2 carbon dioxide / 2-propanol]. The first eluting isomer obtained as a white gum was designated ENT-1 (C56). Yield: 114 mg; this material contained residual ethanol. LCMS m / z 597.1 [M+H]+ Retention time 4.40 minutes (column: Chiral Technologies ChiralCel OD-3, 4.6 x 100 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: 2-propanol containing 0.05% diethylamine; gradient: 5% to 40% B over 4.5 minutes, then maintain at 40% B for 2.5 minutes; flow rate: 2.8 mL / min).

[0850] The second eluting isomer was repurified using SFC [column: Chiral Technologies ChiralCel OD, 5 μm; mobile phase: 55:45 carbon dioxide / (2-propanol containing 0.1% ammonium hydroxide)] to give a colorless gum designated ENT-2 (C57). Yield: 50 mg, 83.8 μmol, 26%. LCMS m / z 597.1 [M+H] + Retention time 4.74 minutes (analytical conditions were the same as those used for C56).

[0851] Step 4: Synthesis of 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X1(8) [from C56]

[0852] A solution of C56 (114 mg, 0.191 mmol) in acetonitrile (10 mL) was treated with an aqueous solution of 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-a]pyrimidine (0.97 M, 394 μL, 0.382 mmol) and the reaction mixture was stirred at room temperature for 23 hours. More aqueous solution of 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-a]pyrimidine (0.97 M, 394 μL, 0.382 mmol) was added and stirring was continued for 6 hours, whereupon the pH was carefully adjusted to 7-8 by addition of 1 M hydrochloric acid. After removal of volatiles in vacuo, purification was performed using reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B: acetonitrile; gradient: 30% to 50% B) to provide 8 as a white solid. Yield: 22.2 mg, 38.1 μmol, 20%. LCMS m / z 583.3 [M+H] + . 1 H NMR (400 MHz, methanol-d 4): δ8.19(d,J=1.4Hz,1H),7.94(dd,J=8.4,1.5Hz,1H),7.77(dd,J=7.7,7.7Hz ,1H),7.64(dd,J=10.6,1.6Hz,1H),7.58(d,J=8.4Hz,1H),7.57(dd,J=8.0,1.5 Hz,1H),6.81-6.75(m,1H),6.75-6.68(m,2H),5.34-5.25(m,1H),4.73(dd,J=1 5.3, 3.0Hz, 1H), 4.67-4.59 (m, 1H), 4.49 (dt, J=9.2, 6.0Hz, 1H), 3.96 (AB quartet, J AB =13.7Hz,Δν AB =41.2Hz,2H),3.06(brd,J=11Hz,1H),2.95(br d,J=11Hz,1H),2.87-2.76(m,1H),2.71(tt,J=12.0,3.9Hz,1H),2.61-2.50(m,1H),2.36-2.21(m,2H),2.06(s,3H),1.95-1.72(m,4H).

[0853] Step 5: Synthesis of 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2(9) [from C57]

[0854] A solution of C57 (50 mg, 84 μmol) in acetonitrile (10 mL) was treated with an aqueous solution of 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-a]pyrimidine (0.97 M, 173 μL, 0.168 mmol). The reaction was stirred for 16 hours at room temperature (about 25 °C), whereupon an additional amount of an aqueous solution of 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-a]pyrimidine (0.97 M, 173 μL, 0.168 mmol) was added and stirring was continued for 29 hours at 25 °C. The reaction mixture was then carefully adjusted to 7 to 8 by the addition of 1 M hydrochloric acid; the resulting mixture was concentrated in vacuo and subjected to reverse phase HPLC (column: Xtimate TM C18, 5 μm; mobile phase A: 0.05% ammonium hydroxide (in water); mobile phase B: acetonitrile; gradient: 27% to 67% B) to give 9 as a white solid. Yield: 18.0 mg, 30.9 μmol, 37%. LCMS m / z 583.3 [M+H] +. 1 H NMR (400 MHz, methanol-d 4 )δ8.36-8.33 (m, 1H), 7.97 (dd, J = 8.5, 1.5 Hz, 1H), 7.78 (dd, J = 7.7, 7.7 Hz, 1H), 7.70-7.63 (m, 2H), 7.57 (dd, J = 8.0, 1.5 Hz, 1H), 6.83-6.76 (m, 1H), 6.76-6.71 (m, 2H), 5.34-5.25 (m, 1H), 4.95-4.85 (m, 1H, assumed; partially obscured by water peak), 4.73 (dd, component of ABX mode, J = 15.3, 2.7 Hz, 1H), 4.68-4.60 (m, 1H), 4.50 (dt, J = 9.2, 6.0 Hz, 1H), 4.02 (AB quartet, J AB =13.8Hz,Δν AB =48.2Hz,2H),3.13(br d,J=11Hz,1H),3.01(br d,J=11.5Hz,1H),2.89-2.78(m,1H),2.78-2.68(m,1H),2.60-2.50(m,1H),2.45-2.30(m,2H),2.07(br s,3H),2.00-1.86(m,2H),1.83(m,2H).

[0855] Example 10

[0856] 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2(10)[from P9]

[0857]

[0858] Step 1: Synthesis of 5-chloro-2-[2-methyl-4-(piperidin-4-yl)-1,3-benzodioxol-2-yl]pyridine, ENT-X2, p-toluenesulfonate (C58) [from P9]

[0859] A solution of P9 (228 mg, 0.529 mmol) in ethyl acetate (2.7 mL) was treated with p-toluenesulfonic acid monohydrate (116 mg, 0.610 mmol) and the reaction mixture was heated at 50° C. for 16 hours. It was then allowed to stir overnight at room temperature, whereupon the precipitate was collected via filtration and rinsed with a mixture of ethyl acetate and heptane (1:1, 2×20 mL) to provide C58 as a white solid. Yield: 227 mg, 0.451 mmol, 85%. LCMS m / z 331.0 ◆ [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 ):δ8.73(d,J=2.4Hz,1H),8.61-8.46(br m,1H),8.35-8.18(br m,1H),8.02(dd,J=8.5,2.5Hz,1H),7.64(d,J=8.5Hz,1H),7.47(d,J=7.8,2H),7.11(d,J=7.8Hz,2H),6.89-6.81(m,2H),6.72(quintet,J=4.0Hz,1H),3.45-3.27(m,2H,assumed;partially obscured by water peak),3.10-2.91(m,3H),2.28(s,3H),2.02(s,3H),1.97-1.80(m,4H).

[0860] Step 2: Synthesis of methyl 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate, DIAST-Y2(C59)[from P9]

[0861] N,N-diisopropylethylamine (0.234 mL, 1.34 mmol) was added to a solution of C58 (225 mg, 0.447 mmol) in acetonitrile (2.2 mL). After stirring this mixture at 45 °C for 5 minutes, P15 (120 mg, 0.407 mmol) was added, and stirring was continued at 45 °C for 16 hours, whereupon P15 (11 mg, 37 μmol) was added again. After stirring for an additional 3 hours, the reaction mixture was treated with water (2.5 mL) and allowed to cool to room temperature. More water (5 mL) was added, and the resulting slurry was stirred for 2 hours, whereupon the solid was collected by filtration and washed with a mixture of acetonitrile and water (15:85, 3 x 5 mL) to give C59 (252 mg) as an off-white solid. 1H NMR analysis showed this material contained some N,N-diisopropylethylamine and it was used directly in the next step. LCMS m / z 589.1◆[M+H] + . 1 H NMR (400 MHz, chloroform-d) 8.61 (d, J = 2.3 Hz, 1H), 8.18 (d, J = 1.5 Hz, 1H), 7.96 (dd, J = 8.5, 1.5 Hz, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.67 (dd, ABX pattern component, J = 8.4, 2.4 Hz, 1H), 7.59-7.51 (m, 1H), 6.82-6.75 (m, 1H), 6.74-6.66 (m, 2H), 5.28-5.19 (m, 1H), 4.75 (dd, ABX pattern component , J=15.3,6.0Hz,1H),4.68(dd, component of ABX pattern, J=15.3,3.4Hz,1H),4.67-4.58(m,1H),4.41(ddd, J=9.1,5.9,5.9Hz,1H),3.95(s,2H),3.95(s,3H),3.07-2.89(m,2H),2.81-2.69(m,2H),2.53-2.41(m,1H),2.37-2.22(m,2H),2.05(s,3H),1.93-1.74(m,4H).

[0862] Step 3: Synthesis of 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2(10) [from P9]

[0863] A suspension of C59 (from the previous step; 250 mg, ≤ 0.407 mmol) in methanol (2 mL) was heated to 40 °C, whereupon aqueous sodium hydroxide solution (1 M; 0.81 mL, 0.81 mmol) was added. After 17 h, the reaction mixture was cooled to room temperature, and the pH was adjusted to 5-6 with 1 M aqueous citric acid solution. The resulting mixture was diluted with water (2 mL), stirred for 2 h, and extracted with ethyl acetate (3 x 5 mL); the combined organic layers were washed with saturated aqueous sodium chloride solution (5 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to provide a foamy solid. This material was taken up in a mixture of ethyl acetate and heptane (1:1, 4 mL), heated to 50 °C, and then allowed to cool and stir overnight. Filtration afforded 10 as a white solid. Yield: 179 mg, 0.311 mmol, 76% (over two steps). LCMS m / z 575.1◆[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ12.73(br s,1H),8.71(d,J=2.5Hz,1H),8.27(d,J=1.5Hz,1H),8.00(dd,J=8.5,2.5Hz,1H),7.8 0(dd,J=8.4,1.6Hz,1H),7.64(d,J=8.4Hz,1H),7.60(d,J=8.5Hz,1H),6.83-6.72(m,3 H), 5.14-5.06 (m, 1H), 4.77 (dd, ABX pattern component, J = 15.2, 7.2 Hz, 1H), 4.63 (dd, ABX pattern component, J = 15.2, 2.8 Hz, 1H), 4.50-4.42 (m, 1H), 4.37 (ddd, J = 9.0, 5.9, 5.9 Hz, 1H), 3.85 (AB quartet, J AB =13.6Hz,Δν AB =71.5Hz,2H),3.01(br d,J=11.2Hz,1H),2.85(br d,J=11.2Hz,1H),2.74-2.57(m,2H),2.47-2.38(m,1H),2.29-2.10(m,2H),2.01(s,3H),1.81-1.64(m,4H).

[0864] Synthesis 10S-1. Example 10, Synthesis of 1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine salt

[0865] Synthesis of 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate, DIAST-X2 (10,1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt) [from P9]

[0866]

[0867] A mixture of 10 (1.54 g, 2.68 mmol) in tetrahydrofuran (10 mL) was treated with an aqueous solution of 2-amino-2-(hydroxymethyl)propane-1,3-diol (Tris, 1.0 M; 2.81 mL, 2.81 mmol). After 24 hours, the reaction mixture was concentrated in vacuo with ethanol (2x50 mL). The residue was treated with ethanol (15 mL). After stirring for 20 hours, the solid was collected by filtration and washed with cold ethanol (5 mL) to obtain 10,1,3-dihydroxy-2-(hydroxymethyl)propane-2-amine salt as a white solid. Yield: 1.41 g, 2.03 mmol, 76%. LCMS m / z 575.3 ◆ [M+H] + . 1 H NMR (600 MHz, DMSO-d 6 )δ8.71(d,J=2.5Hz,1H),8.21(br s,1H),8.00(dd,J=8.5,2.5Hz,1H),7.79(br d,J=8.4Hz,1H),7.60(d,J=8.5Hz,1H),7.57(d,J=8.4Hz,1H),6.82-6.73(m,3H),5.13-5.07(m,1H),4.74(dd,J=15.3,7.2Hz,1H),4.61 (dd,J=15.3,2.9Hz,1H),4.49-4.43(m,1H),4.37(ddd,J=9.0,5.9,5.9Hz,1H),3.93(d,J=13.6Hz,1H),3.75(d,J=13.5Hz,1H),3.01(br d,J=11.3Hz,1H),2.86(br d,J=11.4Hz,1H),2.73-2.59(m,2H),2.48-2.37(m,1H),2.27-2.20(m,1H),2.19-2.12(m,1H),2.01(s,3H),1.82-1.66(m,4H). mp=184℃ to 190℃.

[0868] Synthesis 10S-2. Example 10, Alternative Synthesis of 1,3-Dihydroxy-2-(Hydroxymethyl)propane-2-amine Salt

[0869] A mixture of 10 (8.80 gm, 15.3 mmol) in 2-methyltetrahydrofuran (90 ml) was concentrated in vacuo on a rotary evaporator in a 37°C water bath to reduce the total volume to ~54 ml. Isopropanol (90 ml) was added to the mixture, and the resulting mixture was then concentrated again to a volume of ~54 ml. Isopropanol (135 ml) was added to the mixture, followed by aqueous tris amine (3M; 5.0 ml, 0.98 equiv). The resulting mixture / solution was stirred at ambient temperature; and within ~15 minutes, a solid precipitate began to form. The mixture was then stirred for an additional 5 hours at ambient temperature. The resulting mixture / slurry was cooled to 0°C and the cooled slurry was stirred for an additional approximately 2 hours. The slurry was filtered and washed with cold isopropanol (3 x 15 ml). The collected solids were air dried on the collection funnel for approximately 90 minutes, and then transferred to a vacuum oven to dry overnight. After ~16 hours at 50°C / 23 inHg vacuum (with a small amount of nitrogen vent), 8.66 gm of 10,1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt was obtained as a white solid; 99.8 area% by UPLC (yield: 12.5 mmol, 81%). LCMS and 1 H NMR data were essentially the same as those for the synthesis of 10S-1 shown above.

[0870] Example 10, Obtaining Powder X-ray Diffraction (PXRD) Data of Form A of 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine Salt (also referred to as Form A of the Anhydrous Tris Salt of Compound Example 10)

[0871] The white solid of the tris salt of Example 10 (from Synthesis 10S-1 and Synthesis 10S-2) was subjected to PXRD analysis and found to be a crystalline material (designated as Form A). Powder X-ray diffraction analysis was performed using a Bruker AXSD8 Endeavor diffractometer equipped with a Cu radiation source. The divergence slit was set at 15 mm for continuous illumination. The diffracted radiation was detected by a PSD-LynxEye detector, and the detector PSD was set at 2.99 degrees. The X-ray tube voltage and current intensity were set at 40 kV and 40 mA, respectively. At the Cu wavelength (CuK ā=1.5418λ), in the θ-θ goniometer, using a 0.01 degree step size and a 1.0 second step time, data were collected from 3.0 to 40.0 degrees 2θ. The anti-scatter screen was set to a fixed distance of 1.5 mm. During data collection, the sample was rotated. The sample was prepared by placing it in a silicon low background sample holder and rotating it during collection. Using BrukerDIFFRAC Plus software, data were collected and analyzed by EVADiffrac Plus software. Before peak search, the PXRD data file was not processed. Using the peak search algorithm in the EVA software, the peak selected by threshold 1 was used for preliminary peak assignment. To ensure validity, adjustments were made manually; the output of the automatic assignment was visually inspected, and the peak position was adjusted to the peak maximum. Peaks with relative intensity ≥3% are usually selected. In general, unresolved peaks or peaks consistent with noise are not selected. The typical error associated with the peak position of PXRD specified by USP is up to + / -0.2°2-θ (USP-941). A list of the PXRD diffraction peaks expressed in degrees 2θ and relative intensities (relative intensity ≥ %) of the samples obtained by synthesizing 10S-2 above is provided in Table X2.

[0872] Table X2

[0873]

[0874]

[0875] Embodiment 11

[0876] 1-(2-Methoxyethyl)-2-({4-[2-methyl-2-(pyridin-3-yl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1H-benzimidazole-6-carboxylic acid, formate (11)

[0877]

[0878] This entire synthetic sequence was performed in library format.

[0879] Step 1: Synthesis of methyl 1-(2-methoxyethyl)-2-({4-[2-methyl-2-(pyridin-3-yl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1H-benzimidazole-6-carboxylate (C60)

[0880] A mixture of P14 (44 mg, 100 μmol) and 3-ethynylpyridine (21 mg, 200 μmol) in toluene (800 μL) was treated with sodium bicarbonate (100 μmol) and then with triruthenium dodecacarbonyl (6 mg, 9 μmol). The reaction vial was then capped and shaken at 120°C for 16 hours. Removal of the solvent by concentrator afforded C60 which was used directly in the next step.

[0881] Step 2: Synthesis of 1-(2-methoxyethyl)-2-({4-[2-methyl-2-(pyridin-3-yl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1H-benzimidazole-6-carboxylic acid, formate (11)

[0882] Aqueous sodium hydroxide solution (1.0 M; 200 μL, 200 μmol) was added to a solution of C60 (from the previous step, ≤100 μmol) in a mixture of methanol (400 μL) and tetrahydrofuran (400 μL). The reaction vial was capped and shaken at 80°C for 16 hours, whereupon the mixture was heated to 40°C with 1% ethanol. The reaction mixture was evaporated and purified using reverse phase HPLC (column: Agela Durashell C18, 5 μm; mobile phase A: 0.225% formic acid (in water); mobile phase B: acetonitrile; gradient: 12% to 52% B) to give 11. Yield: 2.2 mg, 4.2 μmol, 4% (over two steps). LCMS m / z 529 [M+H] + Retention time: 2.47 minutes (column: Waters XBridge C18, 2.1 x 50 mm, 5 μm; mobile phase A: 0.0375% trifluoroacetic acid (in water); mobile phase B: 0.01875% trifluoroacetic acid (in acetonitrile); gradient: 1% to 5% B over 0.6 minutes; 5% to 100% B over 3.4 minutes; flow rate: 0.8 mL / minute).

[0883] Example 12

[0884] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[2-(dimethylamino)ethyl]-1H-benzimidazole-6-carboxylic acid (12)

[0885]

[0886] This entire synthetic sequence was performed in library format.

[0887] Step 1: Synthesis of methyl 3-{[2-(dimethylamino)ethyl]amino}-4-nitrobenzoate (C61)

[0888] Methyl 3-fluoro-4-nitrobenzoate (0.2 M solution in N,N-dimethylformamide; 1 mL, 200 μmol) was treated with N,N-dimethylethane-1,2-diamine (18 mg, 200 μmol) and N,N-diisopropylethylamine (78 mg, 600 μmol). The reaction vial was capped and shaken at 50 °C for 16 h, then heated with The reaction mixture was evaporated using a concentrator to afford C61. This material was used directly in the next step.

[0889] Step 2: Synthesis of methyl 4-amino-3-{[2-(dimethylamino)ethyl]amino}benzoate (C62)

[0890] The zinc powder was activated using dilute hydrochloric acid. Methanol (2 mL) was added to C61 (from the previous step, ≤ 200 μmol), followed by an aqueous solution of calcium chloride (1.0 M; 200 μL, 200 μmol) and activated zinc powder (130 mg, 2.0 mmol). The reaction flask was capped and shaken at 70 °C for 16 hours, whereupon the reaction mixture was filtered. The filtrate was concentrated using a concentrator and the residue was taken up in water (2 mL) and then extracted with ethyl acetate (2 x 3 mL). The combined organic layers were evaporated using a concentrator to give C62 (estimated 150 μmol) which was used directly in the next step.

[0891] Step 3: Synthesis of methyl 4-[({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}acetyl)amino]-3-{[2-(dimethylamino)ethyl]amino}benzoate (C63)

[0892] Compound P10 (41 mg, 100 μmol) was added to C62 (from the previous step, approximately 150 μmol), and the mixture was treated with 2-hydroxypyridine 1-oxide and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride in N,N-dimethylacetamide (0.1 M each; 1 mL, 100 μmol each). Then, N,N-diisopropylethylamine (39 mg, 300 μmol) was added, and the reaction vial was capped and shaken at 50°C for 16 hours. Then, the reaction mixture was stirred for 16 hours using a 5% ethanol solution of 1% ethanol. The reaction mixture was concentrated using a centrifuge and purified using preparative thin layer chromatography to provide C63, which was directly carried forward to the next step.

[0893] Step 4: Synthesis of methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[2-(dimethylamino)ethyl]-1H-benzimidazole-6-carboxylate (C64)

[0894] A mixture of acetic acid (500 μL) and C63 (from the previous step, ≤100 μmol) was shaken in a capped vial at 150 °C for 2 h, then The reaction mixture was evaporated using a concentrator. The obtained C64 was directly carried to the next step.

[0895] Step 5: Synthesis of 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[2-(dimethylamino)ethyl]-1H-benzimidazole-6-carboxylic acid (12)

[0896] A solution of C64 (from the previous step, ≤100 μmol) in ethanol (500 μL) was treated with aqueous lithium hydroxide (2.0 M; 500 μL, 1 mmol), and the reaction mixture was shaken in a sealed vial at 50 °C for 2 h. After the pH of the mixture was adjusted to 7 by adding 1.0 M hydrochloric acid, the mixture was heated to 40 °C using The resulting mixture was concentrated by a concentrator and then purified by reverse phase HPLC [column: Agela Durashell C18, 5 μm; mobile phase A: ammonium hydroxide (in water) (pH 10); mobile phase B: acetonitrile; gradient: 25% to 65% B] to give 12. Yield: 7.0 mg, 12 μmol, 12% (over three steps). LCMS m / z 593 [M+H] + Retention time: 2.45 minutes (column: Waters XBridge C18, 2.1 x 50 mm, 5 μm; mobile phase A: 0.0375% trifluoroacetic acid (in water); mobile phase B: 0.01875% trifluoroacetic acid (in acetonitrile); gradient: 10% to 100% B over 4.0 minutes; flow rate: 0.8 mL / minute).

[0897] Embodiment 13

[0898] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-3-(1,3-oxazol-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid (13)

[0899]

[0900] Step 1: Synthesis of methyl 6-[(1,3-oxazol-2-ylmethyl)amino]-5-nitropyridine-2-carboxylate (C65)

[0901] Triethylamine (532 mg, 5.26 mmol) was added to a suspension of 1-(1,3-oxazol-2-yl)methylamine, hydrochloride (236 mg, 1.75 mmol) and 6-chloro-5-nitropyridine-2-carboxylic acid methyl ester (386 mg, 1.78 mmol) in tetrahydrofuran (5 mL). After the reaction mixture was stirred for 14 hours at 25 ° C, it was poured into water (30 mL) and extracted with dichloromethane (2x50 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo; silica gel chromatography (gradient: 0% to 5% methanol (in dichloromethane)) gave C65 as a yellow solid. Yield: 310 mg, 1.11 mmol, 63%. LCMSm / z 278.7 [M+H] + . 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.69-8.61 (br m, 1H), 8.58 (d, J = 8.4 Hz, 1H), 7.65 (d, J = 0.8 Hz, 1H), 7.46 (d, J = 8.4 Hz, 1H), 7.11 (d, J = 1.0 Hz, 1H), 5.07 (d, J = 5.3 Hz, 2H), 3.97 (s, 3H).

[0902] The remainder of this synthetic sequence was performed in library format.

[0903] Step 2: Synthesis of methyl 5-amino-6-[(1,3-oxazol-2-ylmethyl)amino]pyridine-2-carboxylate (C66)

[0904] Aqueous ammonium chloride solution (5.0 M; 400 μL, 2.0 mmol) and activated zinc (131 mg, 2.0 mmol) were added to a solution of C65 (56 mg, 200 μmol) in methanol (2.0 mL). The reaction vial was then capped and shaken at 30°C for 16 hours, whereupon the reaction mixture was filtered. The filtrate was concentrated using a Celite concentrator, then mixed with water (1.0 mL) and extracted with dichloromethane (3 x 1.0 mL); The combined organic layers were concentrated and evaporated to afford C66, which was used directly in the next step.

[0905] Step 3: Synthesis of methyl 5-[({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}acetyl)amino]-6-[(1,3-oxazol-2-ylmethyl)amino]pyridine-2-carboxylate (C67)

[0906] A mixture of P10 (81 mg, 200 μmol) and C66 (from the previous step, ≤ 200 μmol) was mixed with N,N-dimethylacetamide and then treated with N,N-diisopropylethylamine (100 μL, 600 μmol). A solution of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.24 M) and 2-hydroxypyridine 1-oxide (0.1 M) in N,N-dimethylacetamide (1.0 mL, containing 240 μmol of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride and 100 μmol of 2-hydroxypyridine 1-oxide) was added, and the reaction vial was capped and shaken at 50° C. for 16 hours. Then, the reaction mixture was stirred for 16 hours using 4% paraformaldehyde. The volatiles were removed by concentrating and the residue was subjected to preparative thin layer chromatography to afford C67, which was directly carried forward to the next step.

[0907] Step 4: Synthesis of methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-3-(1,3-oxazol-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylate (C68)

[0908] A mixture of acetic acid (1.0 mL) and C67 (from the previous step, ≤ 200 μmol) was shaken at 150 °C for 2 h, then The reaction mixture was evaporated in a concentrator and the obtained C68 was used directly in the next step.

[0909] Step 5: Synthesis of 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-3-(1,3-oxazol-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid (13)

[0910] Aqueous lithium hydroxide (2.0 M; 1.0 mL, 2.0 mmol) was added to a mixture of C68 (from the previous step, ≤ 200 μmol) in tetrahydrofuran (1.0 mL). After adding methanol (500 μL), the reaction vial was capped and shaken at 50 °C for 16 h. After removing the volatiles by concentrator, dimethyl sulfoxide (1.0 mL) was added, and the pH was adjusted to 7-8 using concentrated hydrochloric acid. The resulting mixture was purified using reverse phase HPLC [column: Agela Durashell C18, 5 μm; mobile phase A: ammonium hydroxide (in water) (pH 10); mobile phase B: acetonitrile; gradient: 24% to 64% B] to give 13. Yield: 3.9 mg, 6.5 μmol, 3% (over four steps). LCMS m / z 604 [M+H] + Retention time: 3.14 minutes (column: Waters XBridge C18, 2.1 x 50 mm, 5 μm; mobile phase A: 0.0375% trifluoroacetic acid (in water); mobile phase B: 0.01875% trifluoroacetic acid (in acetonitrile); gradient: 1% to 5% B over 0.6 minutes; 5% to 100% B over 3.4 minutes; flow rate: 0.8 mL / minute).

[0911] Embodiment 14

[0912] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-methyl-1H-benzimidazole-6-carboxylic acid (14)

[0913]

[0914] Step 1: Synthesis of methyl 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-methyl-1H-benzimidazole-6-carboxylate (C69)

[0915] N,N-diisopropylethylamine (683 μL, 3.92 mmol) was added to a mixture of P3 (680 mg, 1.31 mmol) in acetonitrile (5.2 mL); it was stirred at 45 ° C for 5 minutes, whereupon P16 (319 mg, 1.34 mmol) was added. Stirring was continued at 45 ° C for 2.75 hours, and then water (6 mL) was added and stirred for 30 minutes before the reaction mixture was cooled to room temperature. The solid was collected by filtration and washed with a mixture of acetonitrile and water (1:4, 3 x 5 mL) to give C69 as a white solid. Yield: 635 mg, 1.15 mmol, 88%. LCMS m / z 550.1◆[M+H] + . 1H NMR (400MHz, chloroform-d) δ8.15-8.12(m,1H),7.97(dd,J=8.5,1.6Hz,1H),7.74(d,J=8.5Hz,1H),7.50(dd,J=8.2,8.2Hz,1H),7.16-7.07(m,2H),6. 79-6.73(m,1H),6.72-6.65(m,2H),3.98(s,3H),3.96(s,3H),3.88(s, 2H),3.04-2.93(m,2H),2.76-2.66(m,1H),2.37-2.25(m,2H),2.04(br s,3H),1.89-1.78(m,4H).

[0916] Step 2: Synthesis of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-methyl-1H-benzimidazole-6-carboxylic acid (14)

[0917] A mixture of C69 (600 mg, 1.09 mmol) in methanol (11 mL) was heated to 45 ° C and then treated with aqueous sodium hydroxide solution (1 M; 2.2 mL, 2.2 mmol). After 24 hours, the reaction mixture was adjusted to pH 5-6 by adding aqueous citric acid (1 M; 1.1 mL) and then diluted with water (10 mL). The resulting mixture was cooled to room temperature and stirred for 1 hour, whereupon the precipitated solid was collected by filtration and washed with a mixture of methanol and water (1:4; 3 x 5 mL). This provided 14 as a white solid. Yield: 535 mg, 0.998 mmol, 92%. LCMS m / z 536.1◆[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ8.16(d,J=1.5Hz,1H),7.81(dd,J=8.4,1.6Hz,1H),7.64(d,J=8.4Hz,1H),7.59-7.52(m,2H),7.33(dd,J=8.3,2.1Hz,1H),6. 81-6.70(m,3H),3.94(s,3H),3.84(s,2H),3.01-2.91(m,2H),2.70-2.59(m,1H),2.28-2.16(m,2H),2.02(s,3H),1.73(m,4H).

[0918] Examples 15 and 16

[0919] 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, DIAST-X1, trifluoroacetate (15) [from P18 via C71]; and 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, DIAST-X2, trifluoroacetate (16) [from P18 via C72]

[0920]

[0921] Step 1: Synthesis of methyl 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate (C70) [from P18]

[0922] A mixture of P18 (240 mg, 0.699 mmol), C4 (275 mg, 0.800 mmol), cesium carbonate (455 mg, 1.40 mmol), tris(dibenzylideneacetone)dipalladium(0) (40.0 mg, 43.7 μmol), and 1,1'-biphenyl-2,2'-diylbis(diphenylphosphine) (BINAP; 52.2 mg, 83.8 μmol) in toluene (5 mL) was degassed with nitrogen for 5 minutes and then stirred at 90°C for 16 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo; preparative thin layer chromatography (eluent: 1:1 petroleum ether / ethyl acetate) afforded C70 as a mixture of diastereomers as a yellow oil. Yield: 165 mg, 0.272 mmol, 39%. LCMS m / z 628.1◆[M+Na + ].

[0923] Step 2: Isolation of methyl 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate, DIAST-Y1(C71)[from P18]; and methyl 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate, DIAST-Y2(C72)[from P18]

[0924] Separation of stereoisomers on dioxolane in C70 (165 mg, 0.272 mmol) was performed using SFC [column: Chiral Technologies Chiralpak AD, 10 μm; mobile phase: 65:35 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting isomer was designated as DIAST-Y1 (C71), and the second eluting isomer was designated as DIAST-Y2 (C72); both were isolated as white solids.

[0925] C71 yield: 55.0 mg, 90.7 μmol, 33%. LCMS m / z 605.9◆[M+H] + Retention time 4.47 minutes (column: Chiral Technologies Chiralpak AD-3, 4.6 x 100 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: ethanol containing 0.05% diethylamine; gradient: 5% to 40% B over 4.5 minutes, then maintain at 40% B for 2.5 minutes; flow rate: 2.8 mL / min).

[0926] C72 yield: 58.0 mg, 95.7 μmol, 35%. LCMS m / z 628.0◆[M+Na + ]. Retention time 4.88 minutes (analytical conditions were the same as those used for C71).

[0927] Step 3: Synthesis of 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, DIAST-X1, trifluoroacetate (15) [from P18 via C71]

[0928] Aqueous sodium hydroxide solution (3 M; 1.0 mL, 3.0 mmol) was added to a solution of C71 (55.0 mg, 90.7 μmol) in a mixture of methanol (2.0 mL) and tetrahydrofuran (1.0 mL). After the reaction mixture was stirred at 20 °C for 2 hours, the pH was adjusted to 7 by the addition of 1 M hydrochloric acid, and the resulting mixture was extracted with a mixture of dichloromethane and methanol (10:1, 3 x 30 mL). The combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo. Reverse phase HPLC (column: Boston Green ODS, 5 μm; mobile phase A: 0.1% trifluoroacetic acid in water; mobile phase B: acetonitrile; gradient: 10% to 95% B) provided 15 as a white solid. Yield: 35.8 mg, 50.7 μmol, 56%. LCMS m / z 592.3◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4 )δ8.46(s,1H),8.21(d,J=8.6Hz,1H),7.78(d,J=8.6Hz,1H),7.54(dd,J=8.3,8.3Hz,1H),7.16-7.08(m,2H),6.76(dd,J=8.2,8.1Hz,1H),6.55-6.47(m,2H),4.9-4.70(m,2H,assumed;partially obscured by water peak),3.82(t,J=4.9H z,2H),3.66-3.56(m,1H),3.50-3.41(m,1H),3.19-3.09(m,1H),3.15(s,3H),3.08-2.99(m,1H),2.63- 2.57(m,1H),2.27-2.17(m,1H),2.01(s,3H),1.76-1.66(m,2H),1.62-1.50(m,2H),1.35-1.26(m,1H).

[0929] Step 4: Synthesis of 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylic acid, DIAST-X2, trifluoroacetate (16) [from P18 via C72]

[0930] Aqueous sodium hydroxide solution (3 M; 1.0 mL, 3.0 mmol) was added to a solution of C72 (58.0 mg, 95.7 μmol) in a mixture of methanol (2.0 mL) and tetrahydrofuran (1.0 mL). After the reaction mixture was stirred at 20 °C for 2 hours, the pH was adjusted to 7 by the addition of 1 M hydrochloric acid, and the resulting mixture was extracted with a mixture of dichloromethane and methanol (10:1, 3 x 30 mL). The combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo. Reverse phase HPLC (column: Boston Green ODS, 5 μm; mobile phase A: 0.1% trifluoroacetic acid in water; mobile phase B: acetonitrile; gradient: 35% to 95% B) provided 16 as a white solid. Yield: 33.4 mg, 47.3 μmol, 49%. LCMS m / z 592.2◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4 )δ8.53-8.50 (m, 1H), 8.25 (dd, J = 8.6, 1.4 Hz, 1H), 7.80 (br d, J = 8.6 Hz, 1H), 7.57 (dd, J = 8.4, 8.2 Hz, 1H), 7.25 (dd, J = 10.8, 2.0 Hz, 1H), 7.19 (br dd, J = 8.4, 2.1 Hz, 1H), 6.77 (dd, J = 8.2, 8.1 Hz, 1H), 6.55-6.50 (m, 2H), 4.9-4.72 (m, 2H, assumed; partially obscured by water peak), 3.93-3.80 (m, 2H), 3.68-3.58 (m, 1H), 3.41-3.3 (m, 1H, assumed; partially obscured by solvent peak), 3. 25(s,3H),3.22-3.12(m,1H),3.07-2.97(m,1H),2.67(dd,J=8.3,5.8Hz,1H),2.28-2.17 (m,1H),2.01(d,J=1.0Hz,3H),1.86-1.71(m,2H),1.69-1.56(m,2H),1.36-1.26(m,1H).

[0931] Examples 17 and 18

[0932] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylate ammonium, ENT-1(17) and 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylate ammonium, ENT-2(18)

[0933]

[0934] Step 1: Synthesis of methyl 4-[({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}acetyl)amino]-3-{[(1-ethyl-1H-imidazol-5-yl)methyl]amino}benzoate (C73)

[0935] O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (566 mg, 1.49 mmol) was added to a mixture of P19 (340 mg, 1.24 mmol) in N,N-dimethylformamide (10 mL), and the mixture was stirred for 10 minutes at 25°C. Then, a solution of P10 (503 mg, 1.24 mmol) and N,N-diisopropylethylamine (615 μL, 3.53 mmol) in N,N-dimethylformamide (7.7 mL) was added, and the reaction mixture was stirred for 16 hours at 25°C, whereupon it was poured into water (10 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed successively with aqueous ammonium chloride solution (3 x 20 mL) and saturated aqueous sodium chloride solution (2 x 20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. After purification using silica gel chromatography (Gradient: 0% to 5% methanol in ethyl acetate), C73 was obtained as a light brown gum. Yield: 316 mg, 0.477 mmol, 38%. LCMS m / z 662.2◆[M+H] + .

[0936] Step 2: Synthesis of methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylate (C74)

[0937] A solution of C73 (316 mg, 0.477 mmol) in acetic acid (14 mL) was stirred at 55 °C for 16 hours. The solvent was removed under high vacuum and the residue was purified using preparative thin layer chromatography (eluent: 10:1 dichloromethane / methanol) to give C74 as a colorless oil. Yield: 200 mg, 0.310 mmol, 65%. LCMS m / z 644.3◆[M+H] + .

[0938] Step 3: Synthesis of 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylate ammonium, ENT-1(17) and 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylate ammonium, ENT-2(18)

[0939] A mixture of C74 (150 mg, 0.233 mmol) and aqueous sodium hydroxide solution (2 M; 233 μL, 0.466 mmol) in a mixture of methanol (3 mL) and tetrahydrofuran (3 mL) was stirred at 45 °C for 16 h. After the reaction mixture was adjusted to pH 7 by the addition of 1 M hydrochloric acid, it was concentrated in vacuo to give a mixture of 17 and 18. These enantiomers were separated via SFC [column: Chiral Technologies ChiralCel OD, 10 μm; mobile phase: 1:1 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting enantiomer was designated ENT-1 (17) and the second eluting enantiomer was designated ENT-2 (18), both of which were isolated as white solids.

[0940] 17 Yield: 45.0 mg, 69.5 μmol, 30%. LCMS m / z 630.3◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4)δ8.15(br s,1H),8.00(br d,J=8.4Hz,1H),7.81(s,1H),7.72(d,J=8.2Hz,1H),7.56(dd,J=8.3,8.3Hz,1H),7.28(dd,J=10.9,2.0Hz,1H),7.21(dd,J=8.3,2.1Hz,1H),6.77(dd,component of ABC pattern,J=8.0,7.7Hz,1H),6.69(dd,component of ABC pattern,J=7.8,1.2Hz,1H),6.67-6.60(m,2H),5.82(s,2H),4.12(q,J=7.2Hz,2H),3.89(AB quartet,J AB =14.3Hz,Δν AB =6.9 Hz, 2H), 3.00-2.90 (m, 2H), 2.74-2.64 (m, 1H), 2.32-2.21 (m, 2H), 2.02 (s, 3H), 1.82-1.61 (m, 4H), 1.29 (t, J = 7.3 Hz, 3H). Retention time 5.66 minutes (column: Chiral Technologies Chiralpak AD-3, 4.6 x 150 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: methanol containing 0.05% diethylamine; gradient: 5% to 40% B (over 5.5 minutes), then maintained at 40% B for 3.0 minutes; flow rate: 2.5 mL / min).

[0941] 18 Yield: 32.8 mg, 50.7 μmol, 22%. LCMS m / z 630.3◆[M+H] + . 1 H NMR (400 MHz, methanol-d 4 )δ8.15(s,1H),8.00(d,J=8.5Hz,1H),7.81(s,1H),7.72(d,J=8.5Hz,1H),7.56(dd,J=8.3,8.3Hz,1H),7.28(dd,J=10.9,2.0Hz,1H),7.21(dd,J=8.3,2.0Hz,1H),6.77(dd,component of ABC pattern,J=7.8,7.8Hz,1H),6.69(dd,component of ABC pattern,J=7.9,1.2Hz,1H),6.67-6.60(m,2H),5.82(s,2H),4.12(q,J=7.3Hz,2H),3.89(AB quartet,J AB =14.1Hz,Δν AB=7.4 Hz, 2H), 3.01-2.90 (m, 2H), 2.74-2.63 (m, 1H), 2.31-2.21 (m, 2H), 2.02 (s, 3H), 1.82-1.60 (m, 4H), 1.29 (t, J = 7.3 Hz, 3H). Retention time 5.34 minutes (analytical SFC conditions were the same as those used for 17).

[0942] The compounds listed in Table 1 were prepared using procedures similar to those of the examples identified in Table 2 using the appropriate intermediates identified in Table 2. The compounds were purified using methods discussed herein. The final compounds can be isolated as neutral or acid or base salts.

[0943] Table 1: Structures and IUPAC names of Examples 19-102

[0944]

[0945]

[0946]

[0947]

[0948]

[0949]

[0950]

[0951]

[0952]

[0953]

[0954]

[0955]

[0956]

[0957] Table 2: Preparation methods and physicochemical data of Examples 19-102

[0958]

[0959]

[0960]

[0961]

[0962]

[0963]

[0964] ^Areas are hypothetical; peaks partially obscured by water peaks

[0965] ^^Areas are hypothetical; peaks are partially obscured by solvent peaks

[0966] ◆Chlorine isotope pattern was observed.

[0967] 1. The racemic methyl ester [methyl 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate] was separated into its constituent enantiomers via SFC [column: ChiralTechnologies ChiralCel OD-H, 5 μm; mobile phase: 7:3 carbon dioxide / (2-propanol containing 0.1% ammonium hydroxide)]. The first eluting enantiomer ENT-1 (C76) was used in the synthesis of Example 21, and the second eluting enantiomer ENT-2 (C77) was converted to Example 20. C76 retention time: 5.72 minutes (column: Chiral Technologies Chiralpak OD-3, 4.6 x 150 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: 2-propanol containing 0.05% diethylamine; gradient: 5% to 40% B over 5.5 minutes, then maintained at 40% B for 3.0 minutes; flow rate: 2.5 mL / min). C77 retention time: 6.01 minutes (analytical SFC conditions were the same as those used for C76).

[0968] 2. The methyl ester derived from the coupling of C4 and P17 (methyl 2-{6-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]-6-azaspiro[2.5]octan-1-yl}-1-(2-methoxyethyl)-1H-benzimidazole-6-carboxylate) was separated into its constituent stereoisomers in dioxolane via SFC [column: Chiral Technologies Chiralpak AD, 10 μm; mobile phase: 65:35 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting isomer, DIAST-1 (C78), was converted to Example 26; according to 1 Inspection of the H NMR data indicated that this material was the enantiomer of Example 15. The second eluting isomer, DIAST-2 (C79), was used in the synthesis of Example 25; 1Inspection of the HNMR data indicated that this material was the enantiomer of Example 16. C78 retention time: 3.60 minutes (column: Chiral Technologies Chiralpak AD-3, 4.6 x 100 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: ethanol containing 0.05% diethylamine; gradient: 5% to 40% B over 4.5 minutes, then maintained at 40% B for 2.5 minutes; flow rate: 2.8 mL / min). C79 retention time: 3.82 minutes (analytical SFC conditions were the same as those used for C78).

[0969] 3. 4-(4-Bromo-1,3-benzodioxol-2-yl)-3-fluorobenzonitrile was prepared by treating 3-fluoro-4-formylbenzonitrile and 3-bromobenzene-1,2-diol with p-toluenesulfonic acid (in toluene) using a Dean-Stark apparatus to remove water. This material was then reacted with [1-(tert-butoxycarbonyl)piperidin-4-yl](iodo)zinc in the presence of [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) and copper(I) iodide, followed by ester cleavage using p-toluenesulfonic acid to give the desired 3-fluoro-4-[4-(piperidin-4-yl)-1,3-benzodioxol-2-yl]benzonitrile.

[0970] 4. Analytical HPLC conditions. Column: Waters XBridge C18, 2.1 x 50 mm, 5 μm; Mobile phase A: 0.0375% trifluoroacetic acid (in water); Mobile phase B: 0.01875% trifluoroacetic acid (in acetonitrile); Gradient: 10% to 100% B over 4.0 minutes; Flow rate: 0.8 mL / minute.

[0971] 5. Analytical HPLC conditions. Column: Waters XBridge C18, 2.1 x 50 mm, 5 μm; Mobile phase A: 0.0375% trifluoroacetic acid (in water); Mobile phase B: 0.01875% trifluoroacetic acid (in acetonitrile); Gradient: 1% to 5% B over 0.6 minutes; 5% to 100% B over 3.4 minutes; Flow rate: 0.8 mL / min.

[0972] 6. tert-Butyl 4-[2-methyl-2-(pyridin-2-yl)-1,3-benzodioxol-4-yl]-3,6-dihydropyridine-1(2H)-carboxylate was synthesized from 3-bromobenzene-1,2-diol and 2-ethynylpyridine using the procedure described for the synthesis of C12 in Preparation P7. Subsequent hydrogenation over palladium on carbon followed by treatment with hydrogen chloride in ethyl acetate afforded the desired 2-[2-methyl-4-(piperidin-4-yl)-1,3-benzodioxol-2-yl]pyridine, hydrochloride.

[0973] 7. The racemic methyl ester [methyl 1-(2-methoxyethyl)-2-({4-[2-methyl-2-(pyridin-2-yl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1H-benzimidazole-6-carboxylate] was separated into its constituent enantiomers via SFC [column: Chiral Technologies Chiralpak AD, 10 μm; mobile phase: 65:35 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting enantiomer ENT-1 (C80) was used in the synthesis of Example 90, and the second eluting enantiomer ENT-2 (C81) was converted to Example 89. C80 retention time: 4.11 minutes (column: Chiral Technologies Chiralpak AD-3, 4.6 x 100 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: ethanol containing 0.05% diethylamine; gradient: 5% to 40% B over 4.5 minutes, then maintained at 40% B for 2.5 minutes; flow rate: 2.8 mL / min). C81 retention time: 4.62 minutes (analytical SFC conditions were the same as those used for C80).

[0974] 8. The conversion of P8 and P9 to the corresponding cyano substituted derivatives was carried out using the method described for the synthesis of P4 from P2 in the preparation of P4.

[0975] 9. Treatment of 1-(4-chloro-2-fluorophenyl)ethanone with trimethyl orthoformate and p-toluenesulfonic acid provides 4-chloro-1-(1,1-dimethoxyethyl)-2-fluorobenzene, which reacts with 3-bromo-6-fluorobenzene-1,2-diol in the presence of p-toluenesulfonic acid to give 4-bromo-2-(4-chloro-2-fluorophenyl)-7-fluoro-2-methyl-1,3-benzodioxole. This material is converted to the desired tert-butyl 4-[2-(4-chloro-2-fluorophenyl)-7-fluoro-2-methyl-1,3-benzodioxol-4-yl]piperidine-1-carboxylate using the method described in the preparation of P1 for the synthesis of P1 from C2.

[0976] 10. Separation of the stereoisomers of 96 and 97 in dioxolane was performed using SFC [column: Chiral Technologies Chiralpak IG, 5 μm; mobile phase: 3:1 carbon dioxide / (2-propanol containing 0.2% ammonium hydroxide)]. The first eluting isomer was designated as DIAST-1 (96) and the second eluting isomer was designated as DIAST-2 (97).

[0977] 11. Analytical SFC conditions: Column: Chiral Technologies Chiralpak IG, 4.6 x 100 mm, 5 μm; Mobile phase: 7:3 carbon dioxide / (2-propanol containing 0.2% ammonium hydroxide); Flow rate: 1.5 mL / min; Back pressure: 150 bar.

[0978] 12. Synthesis of tert-butyl 2-(chloromethyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylate from tert-butyl 3-fluoro-4-nitrobenzoate and 1-(1,3-oxazol-2-yl)methylamine using the method described for the synthesis of P11. Subsequent reaction with C54 using triethylamine gave tert-butyl 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylate, which was separated into its constituent enantiomers using SFC [column: Chiral Technologies ChiralCel OD-H, 5 μm; mobile phase: 55:45 carbon dioxide / (ethanol containing 0.1% ammonium hydroxide)]. The first eluting enantiomer, ENT-1 (C82), was used for the synthesis of 99, and the second eluting enantiomer, ENT-2 (C83), was converted to 98. C82 retention time: 1.47 minutes (column: Chiral Technologies Chiralpak OD-3, 4.6 x 50 mm, 3 μm; mobile phase A: carbon dioxide; mobile phase B: methanol containing 0.05% diethylamine; gradient: 5% B, 0.2 minutes, then 5% to 40% B over 1.4 minutes, then maintained at 40% B for 1.05 minutes; flow rate: 4 mL / min). C83 retention time: 1.85 minutes (analytical SFC conditions were the same as those used for C82).

[0979] 13. Reaction of 1-bromo-2,3-difluoro-4-nitrobenzene with cuprous (I) cyanide in 1-methylpyrrolidin-2-one at elevated temperature provided 2,3-difluoro-4-nitrobenzonitrile which was subjected to thionyl chloride and methanol to give methyl 2,3-difluoro-4-nitrobenzoate. This material was converted to the desired methyl 2-(chloromethyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylate by using C29 via the method described in the preparation of P11 for the synthesis of P11 from methyl 3-fluoro-4-nitrobenzoate.

[0980] 14. The desired [2-phenyl-4-(piperidin-4-yl)-1,3-benzodioxol-2-yl]methanol was synthesized from 2-oxo-2-phenylethyl acetate by analogy to the method described for the synthesis of C13.

[0981] CHO GLP-1R Clone H6 - Assay 1

[0982] GLP-1R-mediated agonist activity was determined using a cell-based functional assay using a HTRF (homogeneous time-resolved fluorescence) cAMP detection kit (cAMPHI Range Assay Kit; CisBio cat#62AM6PEJ) that measures cAMP levels in cells. The method is a competitive immunoassay between natural cAMP produced by cells and exogenous cAMP labeled with the dye d2. Tracer binding is visualized by a cryptate-labeled mAb anti-cAMP. The specific signal (i.e., energy transfer) is inversely proportional to the cAMP concentration in the standard or experimental sample.

[0983] The sequence encoding human GLP-1R (NCBI Reference Sequence NP_002053.3, including the naturally occurring variant Gly 168Ser) was subcloned into pcDNA3 (Invitrogen) and a cell line stably expressing the receptor was isolated (designated clone H6). 125 I-GLP-1 7-36 Saturation binding analysis (filtration assay procedure) by Perkin Elmer showed that plasma membranes derived from this cell line expressed a high GLP-1R density (K d :0.4nM,B max :1900fmol / mg protein).

[0984] Cells were removed from cryopreservation, resuspended in 40 mL of Dulbecco's phosphate buffered saline (DPBS-Lonza Cat#17-512Q) and centrifuged at 800 x g for 5 minutes at 22° C. The cell pellet was then resuspended in 10 mL of growth medium [DMEM / F12 1:1 mixture containing HEPES, L-Gln, 500 mL (DMEM / F12 Lonza Cat#12-719F), 10% heat-inactivated fetal bovine serum (Gibco Cat#16140-071), 5 mL of 100X Pen-Strep (Gibco Cat#15140-122), 5 mL of 100X L-glutamine (Gibco Cat#25030-081), and 500 μg / mL Geneticin (G418) (Invitrogen #10131035)]. A sample of the cell suspension in 1 mL of growth medium was counted on a Becton Dickinson ViCell to determine cell viability and cell count per ml. The remaining cell suspension was then adjusted with growth medium to deliver 2000 viable cells per well using a Matrix Combi Multidrop reagent dispenser and the cells were dispensed into a white 384-well tissue culture treated assay plate (Corning 3570). The assay plate was then incubated for 48 hours at 37°C in a humidified atmosphere of 5% carbon dioxide.

[0985] Various concentrations of each test compound (in DMSO) were diluted in assay buffer (HBSS with calcium / magnesium (Lonza / BioWhittaker cat#10-527F) / 0.1% BSA (Sigma Aldrich cat#A7409-1L) / 20 mM HEPES (Lonza / BioWhittaker cat#17-737E)) containing 100 μM 3-isobutyl-1-methylxanthine (IBMX; Sigma cat#I5879). The final DMSO concentration was 1%.

[0986] After 48 hours, growth medium was removed from the assay plate wells and cells were treated with 20 μL of serially diluted compounds in assay buffer for 30 minutes at 37°C in a humidified atmosphere 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 plates were then incubated at room temperature and after 60 minutes, changes in HTRF signal were read using an Envision 2104 multilabel plate reader using excitation at 330 nm and emission at 615 and 665 nm. Raw data were converted to nM cAMP by interpolation from a cAMP standard curve (as described in the manufacturer's assay protocol) and relative to saturating concentrations of the full agonist GLP-1 included on each plate. 7-36 The EC values ​​were determined from the agonist dose-response curves analyzed with a curve fitting program using a 4-parameter logistic dose-response equation. 50 Sure.

[0987] CHO GLP-1R Clone C6 - Assay 2

[0988] GLP-1R-mediated agonist activity was determined using a cell-based functional assay using a HTRF (homogeneous time-resolved fluorescence) cAMP detection kit (cAMPHI Range Assay Kit; Cis Bio cat#62AM6PEJ) that measures cAMP levels in cells. The method is a competitive immunoassay between natural cAMP produced by cells and exogenous cAMP labeled with the dye d2. Tracer binding is visualized by a cryptate-labeled mAb anti-cAMP. The specific signal (i.e., energy transfer) is inversely proportional to the cAMP concentration in the standard or experimental sample.

[0989] The sequence encoding human GLP-1R (NCBI Reference Sequence NP_002053.3, including the naturally occurring variant Leu260Phe) was subcloned into pcDNA5-FRT-TO and expressed using Flp-In TM T-Rex TM A clonal CHO cell line stably expressing low receptor density was isolated using the ELISA system as described by the manufacturer (ThermoFisher). 125 Saturation binding analysis (filtration assay procedure) of I-GLP-1 (Perkin Elmer) showed that plasma membranes derived from this cell line (designated clone C6) expressed a low density of GLP-1Rs (K d :0.3nM,B max:240 fmol / mg protein), relative to clone H6 cell line.

[0990] Cells were removed from cryopreservation, resuspended in 40 mL of Dulbeco's Phosphate Buffered Saline (DPBS - Lonza Cat #17-512Q) and centrifuged at 800 xg for 5 minutes at 22°C. DPBS was aspirated and the cell pellet was resuspended in 10 mL of complete growth medium (DMEM:F12 1:1 mixture containing HEPES, L-Gln, 500 mL (DMEM / F12 Lonza Cat#12-719F), 10% heat-inactivated fetal bovine serum (Gibco Cat#16140-071), 5 mL of 100X Pen-Strep (Gibco Cat#15140-122), 5 mL of 100X L-glutamine (Gibco Cat#25030-081), 700 μg / mL hygromycin (Invitrogen Cat#10687010) and 15 μg / mL blasticidin (Gibco Cat#R21001)). A sample of the cell suspension in 1 mL of growth medium was counted on a Becton Dickinson ViCell to determine cell viability and cell count per ml. The remaining cell suspension was then adjusted with growth medium to deliver 1600 viable cells per well using a Matrix CombiMultidrop reagent dispenser and the cells were dispensed into white 384-well tissue culture treated assay plates (Corning 3570). The plates were then incubated at 37°C in a humidified atmosphere (95% O 2 ,5% CO 2 ) and incubate the assay plates for 48 hours.

[0991] Various concentrations of test compounds (in DMSO) were diluted in assay buffer [HBSS with calcium / magnesium (Lonza / BioWhittaker cat#10-527F) / 0.1% BSA (Sigma Aldrich cat#A7409-1L) / 20 mM HEPES (Lonza / BioWhittaker cat#17-737E)] containing 100 μM 3-isobutyl-1-methylxanthine (IBMX; Sigma cat#I5879). The final DMSO concentration in the compound / assay buffer mixture was 1%.

[0992] After 48 hours, the growth medium was removed from the assay wells and incubated at 37°C in a humidified atmosphere (95% O 2 ,5%CO 2), cells were treated with 20 μL of serially diluted compounds in assay buffer for 30 min. After the 30 min 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 plates were then incubated at room temperature and after 60 min, changes in the HTRF signal were read using an Envision 2104 multilabel plate reader using excitation at 330 nm and emission at 615 and 665 nm. Raw data were converted to nM cAMP by interpolation from a cAMP standard curve (as described in the manufacturer's assay protocol), and the percent effect was determined relative to a saturating concentration of the full agonist GLP-1 (1 μM) included in each plate. EC values ​​were calculated from agonist dose-response curves analyzed with a curve fitting program using a 4-parameter logistic dose-response equation. 50 Sure.

[0993] In Table 3, the assay data are presented to two (2) significant figures as geometric means (EC 50 s) and the arithmetic mean value (Emax). Blanks mean that there are no data for the example or Emax is not calculated.

[0994] Table 3: Biological activities of Examples 1-102

[0995]

[0996]

[0997]

[0998]

[0999] *Tested as ammonium and trifluoroacetate salts

[1000] **Tested as ammonium and 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine (Tris) salts, and free acid***Tested as ammonium salt and free acid

[1001] ****Tested as formate and free acid.

[1002] All patents, patent applications, and references cited herein are hereby incorporated by reference in their entirety.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt thereof, wherein R is F, Cl, or -CN; p is 0 or 1; Ring A is phenyl or 6-membered heteroaryl; m is 0, 1, 2, or 3; Each R 1 Independently selected from: halogen, -CN, -C 1-3 Alkyl, or -OC 1-3 Alkyl, where C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group is substituted with 0 to 3 F atoms; R 2 Is H or -C 1-3 Alkyl, wherein the alkyl group is substituted with 0 to 1 OH groups; Each R 3 are independently F, -OH, -CN, -C 1-3 Alkyl, -OC 1-3 Alkyl, or -C 3-4 Cycloalkyl, or 2 R 3 Can cyclize together to form -C 3-4 Spirocycloalkyl, where, if valence permits, C 1-3 Alkyl and OC 1-3 The alkyl, cycloalkyl, or spirocycloalkyl group of the alkyl group may be substituted by 0 to 3 F atoms and by 0 to 1 -OH groups; q is 0, 1, or 2; XL is N-CH2, CHCH2, or cyclopropyl; Y is CH or N; R 4 Yes-C 1-3 Alkyl, -C 0-3 Alkylene-C 3-6 Cycloalkyl, -C 0-3 Alkylene-R 5 , or -C 1-3 Alkylene-R 6 , wherein if valence permits, the alkyl group may be substituted by 0 to 3 substituents independently selected from 0 to 3 F atoms and 0 to 1 substituent selected from -C 0-1 Alkylene -CN, -C 0-1 Alkylene-OR O 、-SO2-N(R N )2、-C(O)-N(R N )2, -N(C=O)(R N ), and -N(R N )2 is substituted with a substituent; and Wherein, if valence permits, the alkylene and cycloalkyl groups may be independently substituted with 0 to 2 substituents independently selected from 0 to 2 F atoms and 0 to 1 substituent selected from -C 0-1 Alkylene -CN, -C 0-1 Alkylene-OR O , and -N(R N ) 2 is substituted by a substituent; R 5 is a 4- to 6-membered heterocycloalkyl group, wherein the heterocycloalkyl group may be substituted, if valence permits, with 0 to 2 substituents independently selected from: 0 to 1 oxo (=O), 0 to 1 -CN, 0 to 2 F atoms, and 0 to 2 independently selected from -C 1-3 Alkyl and -OC 1-3 Alkyl substituents, where valence permits, C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group may be substituted with 0 to 3 substituents independently selected from the following: 0 to 3 F atoms, 0 to 1 -CN, and 0 to 1 -OR O ; R 6 is a 5- to 6-membered heteroaryl group, wherein, if valence permits, the heteroaryl group may be substituted with 0 to 2 substituents independently selected from: 0 to 2 halogens, 0 to 1 selected from -OR O and -N(R N )2, and 0 to 2 -C 1-3 Alkyl, wherein if valence permits, the alkyl may be substituted with 0 to 3 substituents independently selected from: 0 to 3 F atoms, and 0 to 1 -OR O ; Each R O are independently H, or -C 1-3 Alkyl, where C 1-3 The alkyl group may be substituted by 0 to 3 F atoms; Each R N are independently H, or -C 1-3 alkyl; Z 1 , Z 2 , and Z 3 Each is -CR Z ,or Z 1 , Z 2 , and Z 3 One of them is N and the other two are -CR Z ;as well as Each R Z are independently H, F, Cl, or -CH3.

2. The compound of claim 1, wherein the compound is a compound of formula II or a pharmaceutically acceptable salt thereof, wherein R is F; p is 0 or 1; Ring A is phenyl or pyridyl; m is 0, 1, or 2; Each R 1 Independently selected from: halogen, -CN, -C 1-3 Alkyl, or -OC 1-3 Alkyl, where C 1-3 Alkyl and OC 1-3 The alkyl group of the alkyl group is substituted with 0 to 3 F atoms; R 2 is H or CH3; XL is N-CH2, or cyclopropyl; Y is CH or N; Z 3 Yes-CR Z or N; and R Z It is H, F, Cl, or -CH3.

3. The compound of claim 1 or claim 2, wherein the compound is a compound of formula III or a pharmaceutically acceptable salt thereof, wherein Ring A is phenyl or pyridyl; m is 0, 1, or 2; Each R 1 Independently selected from F, Cl, or -CN; R 2 is H or CH3; and Y is CH or N.

4. The compound of any one of claims 1 to 3, wherein R4 is -CH2-R 5 , where R 5 is a 4- to 5-membered heterocycloalkyl group, wherein, if valence permits, the heterocycloalkyl group may be substituted with 0 to 2 substituents independently selected from: 0 to 2 F atoms, and 0 to 1 substituents selected from -OCH3 and -CH2OCH3; or a pharmaceutically acceptable salt thereof.

5. The compound of any one of claims 1 to 3, wherein R4 is -CH2-R 6 , where R 6 is a 5-membered heteroaryl, wherein, if valence permits, the heteroaryl may be substituted with 0 to 2 substituents independently selected from: 0 to 2 halogens, wherein the halogens are independently selected from F and Cl, 0 to 1 -OCH3, and 0 to 1 -CH3, -CH2CH3, -CF3, or -CH2CH2OCH3; or a pharmaceutically acceptable salt thereof.

6. The compound of any one of claims 1 to 5, wherein R 2 is H, or a pharmaceutically acceptable salt thereof.

7. The compound of claim 6, wherein the compound is 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, or a pharmaceutically acceptable salt thereof.

8. The compound of claim 6, wherein the compound is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, or 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, or a pharmaceutically acceptable salt thereof.

9. A compound which is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, or a pharmaceutically acceptable salt thereof, wherein the salt is a tris salt.

10. A compound which is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid as a free acid.

11. A compound which is or a pharmaceutically acceptable salt thereof.

12. The compound of any one of claims 1 to 5, wherein R 2 is CH3, or a pharmaceutically acceptable salt thereof.

13. The compound of claim 12, wherein the compound is 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-3-(1,3-oxazol-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-4-ylmethyl)-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(pyridin-3-ylmethyl)-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-5-ylmethyl)-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-1,2,3-triazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-chloro-2-fluorophenyl)-7-fluoro-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylic acid; or 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or a pharmaceutically acceptable salt thereof.

14. The compound of claim 12, wherein the compound is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or a pharmaceutically acceptable salt thereof.

15. A compound which is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, or a pharmaceutically acceptable salt thereof, wherein the salt is a tris salt.

16. A compound which is 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid as a free acid.

17. A compound which is or a pharmaceutically acceptable salt thereof.

18. The compound of claim 12, wherein the compound is 2-({4-[(2S)-2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid; or a pharmaceutically acceptable salt thereof.

19. The compound of claim 12, wherein the compound is 2-({4-[(2R)-2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid, or a pharmaceutically acceptable salt thereof.

20. A compound which is the following compound as a free acid 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid.

21. A compound which is 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid; or a pharmaceutically acceptable salt thereof, wherein the salt is a tris salt.

22. A compound which is 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2: or a pharmaceutically acceptable salt thereof.

23. A crystalline form of anhydrous 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt of 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid (Form I), having a powder X-ray diffraction pattern (CuKα radiation) comprising at least two characteristic peaks selected from 3.7±0.2°; 7.3±0.2°; 8.5±0.2°; 10.1±0.2°; 14.7±0.2°; and 16.9±0.2° in terms of 2θ.

24. The crystalline form of claim 23, which has a powder X-ray diffraction pattern comprising peaks, which are at least three characteristic peaks selected from 3.7±0.2°; 7.3±0.2°; 8.5±0.2°; 10.1±0.2°; 14.7±0.2°; and 16.9±0.2° in terms of 2θ.

25. The crystalline form of claim 23, which has a powder X-ray diffraction pattern comprising peaks, wherein the peaks are at least four characteristic peaks selected from 3.7±0.2°; 7.3±0.2°; 8.5±0.2°; 10.1±0.2°; 14.7±0.2°; and 16.9±0.2° in terms of 2θ.

26. The crystalline form of claim 23, which has a powder X-ray diffraction pattern comprising peaks, wherein the peaks are at least five characteristic peaks selected from 3.7±0.2°; 7.3±0.2°; 8.5±0.2°; 10.1±0.2°; 14.7±0.2°; and 16.9±0.2° in terms of 2θ.

27. The crystalline form of claim 23, which has a powder X-ray diffraction pattern comprising characteristic peaks at 3.7±0.2° and 7.3±0.2° in terms of 2θ.

28. The crystalline form of claim 23, having a powder X-ray diffraction pattern comprising peaks at 3.7±0.2°; 7.3±0.2° and 14.7±0.2° in terms of 2θ.

29. The crystalline form of claim 27 or 28, wherein the X-ray powder diffraction pattern further comprises at least one peak selected from 8.5±0.2°; 10.1±0.2°; and 16.9±0.2° in terms of 2θ.

30. The crystalline form of claim 29, having a powder X-ray diffraction pattern comprising peaks at 3.7±0.2°; 7.3±0.2°; 14.7±0.2°; and 16.9±0.2° in terms of 2θ.

31. The crystalline form of claim 30, having a powder X-ray diffraction pattern comprising peaks at 3.7±0.2°; 7.3±0.2°; 8.5±0.2°; 10.1±0.2°; 14.7±0.2°; and 16.9±0.2° in terms of 2θ.

32. The crystalline form of any one of claims 23 to 31 , having a powder X-ray diffraction pattern substantially as shown in Figure 1.

33. A crystalline form (Form A) of the anhydrous 1,3-dihydroxy-2-(hydroxymethyl)propan-2-amine salt of 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2, having a powder X-ray diffraction pattern (CuKα radiation) comprising at least two characteristic peaks selected from 7.7±0.2°; 15.2±0.2°; 15.7±0.2° and 17.6±0.2° in terms of 2θ.

34. The crystalline form of claim 33, which has a powder X-ray diffraction pattern comprising at least three characteristic peaks selected from 7.7±0.2°; 15.2±0.2°; 15.7±0.2° and 17.6±0.2° in terms of 2θ.

35. The crystalline form of claim 33, which has a powder X-ray diffraction pattern comprising characteristic peaks at 7.7±0.2° and 17.6±0.2° in terms of 2θ.

36. The crystalline form of claim 33, having a powder X-ray diffraction pattern comprising peaks at 7.7±0.2°; 15.2±0.2°; and 17.6±0.2° in terms of 2θ.

37. The crystalline form of claim 33, having a powder X-ray diffraction pattern comprising peaks at 7.7±0.2°; 15.2±0.2°; and 15.7±0.2° in terms of 2θ.

38. The crystalline form of claim 33, having a powder X-ray diffraction pattern comprising peaks at 7.7±0.2°; 15.2±0.2°; 15.7±0.2° and 17.6±0.2° in terms of 2θ.

39. The crystalline form of any one of claims 33 to 38, having a powder X-ray diffraction pattern substantially as shown in Figure 2.

40. A pharmaceutical composition comprising a compound according to any one of claims 1 to 22 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

41. A method for treating cardiovascular metabolic and related diseases or disorders, the method comprising administering to a mammal in need of such treatment a therapeutically effective amount of a compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein the disease or disorder is selected from the group consisting of: T1D, T2DM, prediabetes, idiopathic T1D, LADA, EOD, YOAD, MODY, malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, nephropathy, diabetic retinopathy, adipocyte dysfunction, visceral fat deposition, sleep apnea, obesity, eating disorders, weight gain due to use of other agents, excessive sugar cravings, dyslipidemia, hyperinsulinemia, NAFLD, NASH, fibrosis, Cirrhosis, hepatocellular carcinoma, cardiovascular disease, atherosclerosis, coronary artery disease, peripheral vascular disease, hypertension, endothelial dysfunction, impaired vascular compliance, congestive heart failure, myocardial infarction, stroke, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidemia, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, macular degeneration, cataract, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina, thrombosis, atherosclerosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, impaired fasting glucose condition, hyperuricemia, gout, erectile dysfunction, skin and connective tissue disorders, psoriasis, foot ulcers, ulcerative colitis, high apo Prevention or treatment of beta-lipoproteinemia, Alzheimer's disease, schizophrenia, impaired cognitive function, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel syndrome, polycystic ovary syndrome, and treatment of addiction.

42. A pharmaceutical composition comprising the crystalline form of any one of claims 23 to 39 and a pharmaceutically acceptable excipient.

43. A method of treating cardiovascular metabolic and related diseases or disorders, the method comprising administering to a mammal in need of such treatment a therapeutically effective amount of a crystalline form of any one of claims 23 to 39, wherein the disease or disorder is selected from the group consisting of: T1D, T2DM, prediabetes, idiopathic T1D, LADA, EOD, YOAD, MODY, malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, nephropathy, diabetic retinopathy, adipocyte dysfunction, visceral fat deposition, sleep apnea, obesity, eating disorders, weight gain due to use of other agents, excessive sugar cravings, dyslipidemia, hyperinsulinemia, NAFLD, NASH, fibrosis, cirrhosis, hepatic cell cancer, cardiovascular disease, atherosclerosis, coronary artery disease, peripheral vascular disease, hypertension, endothelial dysfunction, impaired vascular compliance, congestive heart failure, myocardial infarction, stroke, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipidemia, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, macular degeneration, cataract, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina, thrombosis, atherosclerosis, transient ischemic attack, vascular restenosis, abnormal glucose metabolism, impaired fasting glucose condition, hyperuricemia, gout, erectile dysfunction, skin and connective tissue disorders, psoriasis, foot ulcers, ulcerative colitis, high apo Prevention or treatment of beta-lipoproteinemia, Alzheimer's disease, schizophrenia, impaired cognitive function, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel syndrome, polycystic ovary syndrome, and treatment of addiction.

Images