Pyrrolidine-2-carboxamide derivatives as prostaglandin E2 receptor 4 (EP4) agonists for treatment of gastrointestinal and lung diseases

Abstract

The present invention relates to compounds of formula I as prostaglandin E2 receptor 4 (EP4) agonists for use in methods of treating gastrointestinal and pulmonary diseases or conditions. The present disclosure provides exemplary compounds and pharmacological data. # imgabs0 #

Description

Technical Field

[0001] The present application relates to novel compounds and their use as prostaglandin E2 receptor 4 (EP4) agonists. The compounds described herein can be used to treat or prevent diseases in which the EP4 receptor is involved. The present application also relates to pharmaceutical compositions comprising these compounds, and the preparation and use of these compounds and compositions in preventing or treating such diseases in which the EP4 receptor is involved. Background Art

[0002] Prostaglandins (comprising prostaglandins and thromboxanes) are metabolic derivatives of arachidonic acid that play a key role in cellular physiological functions.Arachidonic acid is the indispensable component of the membrane phospholipids released by the activity of phospholipase A2 (phospholipase A2, PLA2).The biosynthesis of prostaglandins is mediated by cyclooxygenase (cyclooxygenase, COX), and the cyclooxygenase catalysis arachidonic acid is converted into unstable intermediate (PGH2) and causes the generation of prostaglandins including PGE2.PGE2 represents the most widely produced prostaglandins, and its activity is mediated by acting on 4 kinds of different receptor subtypes EP1-4 of functions.

[0003] EP receptors belong to the G protein-coupled receptor (GPCR) family of integral membrane proteins with seven transmembrane domains. Such receptors can be broadly classified according to their signaling pathways: i) Gs-coupled receptors (adenylate cyclase activation and cyclic adenosine monophosphate (cAMP) production), EP2 and EP4; ii) Gq-coupled receptors (PLC activation), EP1; and iii) Gi-coupled receptors (adenylate cyclase inhibition), EP3.

[0004] The EP4 receptor signals through Gs and positively couples to adenylate cyclase to increase cAMP levels. The receptor was originally described in 1993, where an EP2-like receptor was identified that positively couples to adenylate cyclase but does not bind butaprost (A Honda et al. J. Biol. Chem. 1993, 268, 7759-7762).

[0005] EP4 receptors play a key role in a variety of physiological functions, including gastrointestinal homeostasis, regulation of vascular tone, renal function, inflammation, fever, and carcinogenesis. The potent biological effects of PGE2 have stimulated interest in developing subtype-selective EP4 agonists and antagonists for the treatment of a wide range of indications.

[0006] Functional gastrointestinal diseases (FGIDs), including chronic constipation, are common gastrointestinal disorders encountered by primary care physicians and gastroenterologists. The prevalence of chronic constipation is 1% to 8% and can negatively impact quality of life (QoL), leading to a serious social and economic burden. Chronic constipation can cause discomfort to patients and affect their daily lives. Symptoms include hard and lumpy stools, straining during defecation, and a feeling of incomplete defecation. The use of laxatives and stool softeners remains high, although many patients do not gain substantial benefit. There is still a need for a treatment that can provide complete and sustainable symptom relief.

[0007] Intestinal mucosa plays an important role in the homeostasis of anion and body fluid maintenance. These functions are mediated by coordinated ion transport via membrane-bound transporters and channels on the top and basolateral membranes of intestinal epithelial cells. PGE2 is a recognized secretagogue that can directly promote intestinal epithelial cells to secrete chloride. The secretory effect of PGE2 is partly mediated by EP4 receptors, which can stimulate anion chloride secretion through intestinal mucosa. Lubiprostone (lubiprostone) (bicyclic fatty acid derivatives of PGE1) approved clinically for the treatment of chronic constipation and IBS-C has been shown to promote fluid secretion and gastrointestinal motility by activating prostaglandin (EP4) receptors. EP4 selective agonists can have therapeutic value in promoting the intestinal fluid homeostasis in chronic constipation disorders.

[0008] Inflammatory bowel disease (IBD) is a chronic debilitating gastrointestinal disorder, including ulcerative colitis and Crohn's disease. Patients with IBD usually show symptoms including diarrhea, abdominal pain, weight loss, rectal bleeding and fever. Clinical management includes strategies for controlling the abnormal dysregulated immune response of the intestinal mucosa. A wide range of agents are used to induce and maintain remission, depending on the severity of the disease. These include aminosalicylates, corticosteroids, immunosuppressants, antibiotics and biologics. This can be given in a stepwise approach, wherein treatment is strengthened according to the severity and progression of the disease. However, it is clear that some patients are refractory to drug therapy, or cannot tolerate drug therapy due to systemic side effects. Only a portion of patients achieve long-term remission due to current treatment strategies, which indicates the need for improved treatment.

[0009] Intestinal barrier dysfunction plays a key pathogenic role in IBD, and there is interest in the development of agents that restore barrier function (mucosal healing). EP4 is expressed in many cell types, including gastrointestinal (GI) epithelial cells, lamina propria mononuclear cells, and colonic innervating sensory neurons, and can provide benefits in inhibiting inappropriate mucosal immune responses and protecting the GI mucosal barrier. PGE2 signaling through EP4 promotes cell differentiation into wound-related epithelial cell phenotypes, which is important for wound repair (Miyoshi, H. et al. EMBO J. 2017, 36, 5--24). Administration of EP4 agonists provides benefits in a chemically induced colitis model (Kabashima, K. et al. J. Clin. Invest. 2002, 109, 883–893; Watanabe, Y. et al. Eur. J. Pharmacol. 2015, 754, 179–189; Nitta, M. et al. Scand. J. Immunol. 2002, 56, 66–75), while mutant mice lacking EP4 develop severe dextran sulfate sodium-induced colitis, characterized by impaired mucosal barrier function, increased epithelial cell loss, crypt damage, and increased immune cell infiltration (Kabashima, K. et al. J. Clin. Invest. 2002, 109, 883–893). In a small PhII study, the EP4 agonist ONO-4819CD was evaluated in patients with mild to moderate ulcerative colitis refractory to 5-ASA (Nakase, H. et al. Inflamm. Bowel Dis. 2010, 16, 731-733). Although the study did not demonstrate efficacy, patients treated with ONO-4819CD showed signs of improved disease activity index (DAI) scores and histological scores. These data support the potential clinical benefits of EP4 agonist treatment in IBD.

[0010] Asthma and chronic obstructive pulmonary disease are inflammatory diseases of the airways characterized by airflow limitation. It is estimated that about 300 million people suffer from asthma and represent the most common chronic diseases in children. Although progress has been made in asthma management, the disease of a considerable number of patients is not controlled, which can lead to death and morbidity. Still need to achieve effective control of asthma symptoms and reduce the treatment of future exacerbation risks for long-term management. Known PGE2 has bronchodilator and anti-inflammatory effects in the airway smooth muscle isolated from rodents and people. It has been shown that the PGE2 inhaled is beneficial to the inflammation and airway diameter (calibre) of the patient suffering from chronic bronchitis and asthma. However, PGE2 also induces reflex cough, potentially through EP3 receptor-mediated upper airway stimulation. This makes efforts to find EP receptor selective agents for the treatment of airway diseases. Interestingly, it is reported that there are key species differences in the receptor subtypes responsible for regulating airway smooth muscle relaxation. In guinea pigs, monkeys and mice, EP2 agonists can induce relaxation of airway smooth muscle, while in humans this is mediated through the EP4 receptor (Buckley, J. et al. Thorax 2011, 66, 1029-1035). Selective EP4 receptor agonists may have potential therapeutic value in airway diseases.

[0011] It has been shown that EP4 receptors play a role in blood pressure regulation. EP4 is expressed on smooth muscle and endothelial cells, and may induce vasodilation by nitric oxide (nitric oxide, NO) mediated by endothelial nitric oxide synthase (endothelial nitric oxide synthase, eNOS). It has been shown that PGE2 relaxes smooth muscle in aortic rings in a dose-dependent manner, and this effect is eliminated in EP4 knockout mice. In dogs anesthetized by halothane, EP4 selective agonist ONO-AE1-329 induces vasodepressor response (Honda, A. et al. Eur. J. Pharmacol. 2016, 775, 130–137) and similarly, hypotension is reported as one of the adverse drug reactions of IBD patients receiving ONO-4819CD (Nakase, H. et al. Inflamm. Bowel Dis. 2010, 16, 731–733).

[0012] EP4 agonists that can be used to treat a range of gastrointestinal and airway disorders without cardiovascular side effects may have potential therapeutic value. In particular, EP4 agonists that can be used to treat a range of gastrointestinal disorders without cardiovascular side effects may have potential therapeutic value. There is a need to discover safe and effective EP4 selective agents. Summary of the invention

[0013] The present invention provides compounds having activity as prostaglandin E2 receptor 4 (EP4) agonists.

[0014] In one aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0015]

[0016] in:

[0017] A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R', S(O)2R', S(O)2OR', SO2N(R')2, C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0018] Ring B is aryl or heteroaryl;

[0019] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0020] R 1 and R 2 Each independently is H, C 1-6 Alkyl, C 1-6 Alkoxy, or R 1 and R 2 Together with the carbon atom to which they are attached, they form C 3-6 Cycloalkane-1,1-diyl;

[0021] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0022] R 4 It is H, C 1-6 Alkyl, halogen, CN, NO2 or OR';

[0023] R 5 Is H or C 1-6 Alkyl; or R 4 and R 5 Together with the pyrrolidine ring to which they are attached, they form C 1-6 Alkylene linker;

[0024] R 6 It is H, C 1-6 alkyl, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkoxy, halogen, CN, NO2, OR', COOR' or C(O)R',

[0025] R7 Is OR', OC(O)R', OC(O)OR', CO2R', CON(R')2, SO2N(R')2, SO2R', OSO2R', or OSO2N(R')2;

[0026] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0027] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0028] n and m are each independently 0, 1, 2 or 3;

[0029] wherein at each occurrence, alkyl, alkylene and cycloalkyl are each optionally and independently substituted with OH', SH, CN, NO2, COOH, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0030] wherein heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO2, COOR', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0031] In another aspect, the present invention provides a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0032]

[0033] in:

[0034] A is selected from:

[0035]

[0036] U, V, W and Z are each independently selected from CH, COH, N or N + -O - , wherein at least three of U, V, W and Z are CH;

[0037] X and Y are each independently selected from CH, CF, COH or N;

[0038] R 1 is H, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl, or R 1 With R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring;

[0039] R2 is H, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl, or R 2 With R 1 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring;

[0040] R 3 is H, OH or F;

[0041] R 4 H, OH, F or R 5 connected to form a CH2 bridge;

[0042] R 5 H or R 4 connected to form a CH2 bridge;

[0043] R 6 is H, OH, CN, halogen, C optionally substituted by 1 to 3 fluorine atoms 1-3 alkoxy, or C optionally substituted by 1 to 3 fluorine atoms 1-3 alkyl;

[0044] R 7 is OH, CO2H, CONH2, SO2NH2 or OSO2NH2; and

[0045] R 9 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring.

[0046] In another aspect, the present invention includes a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable excipient.

[0047] In another aspect, the invention includes a kit comprising a compound described herein and at least one additional therapeutic agent selected from the group consisting of an aminosalicylate, a corticosteroid, an immunomodulator, and combinations thereof.

[0048] In another aspect, the present invention includes a compound described herein, a composition described herein, or a kit described herein for use as a medicament.

[0049] In another aspect, the invention includes a compound described herein, a composition described herein, or a kit described herein for use in treating a disease mediated by the EP4 receptor.

[0050] In another aspect, the invention includes a method of modulating EP4 receptor agonist activity in a biological sample, the method comprising contacting the EP4 receptor with a compound or composition described herein.

[0051] In another aspect, the invention includes a method of treating a disease mediated by the EP4 receptor, comprising administering to a patient in need thereof a compound or composition described herein.

[0052] The compounds herein can be used as EP4 receptor agonists. The compounds herein can selectively act on the EP4 receptor. The compounds can be used to prepare compositions or drugs. The compounds, compositions or drugs can be used to treat, prevent, improve, control or reduce the risk of diseases or conditions involving the EP4 receptor. The compounds, compositions or medicaments can be used to treat, prevent, ameliorate, control or reduce the risk of gastrointestinal disorders and conditions, including but not limited to constipation disorders, constipation-dominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastric and intestinal damage, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrheal diseases, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease and ischemic colitis.

[0053] The compounds, compositions or drugs can also be used to treat, prevent, improve, control or reduce the risk of lung diseases and disorders, such as chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperresponsive airway disorders and idiopathic pulmonary fibrosis. DETAILED DESCRIPTION

[0054] The present invention relates to novel compounds. The present invention also relates to the use of novel compounds as EP4 receptor agonists. The present invention also relates to the use of novel compounds in the preparation of medicaments used as EP4 receptor agonists, and treatment methods comprising administering the compounds of the present invention as EP4 receptor agonists.

[0055] The compounds of formula I can be used to treat, prevent, improve, control or reduce the risk of diseases or conditions in which the EP4 receptor is involved. The compounds of formula I can be used to treat, prevent, improve, control or reduce the risk of gastrointestinal disorders and conditions, including but not limited to constipation disorders, constipation-dominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastric and intestinal damage, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrheal diseases, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease and ischemic colitis.

[0056] The compounds of Formula I may also be used to treat, prevent, ameliorate, control or reduce the risk of lung diseases and conditions, such as chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic essential cough, hyperresponsive airway disorders and idiopathic pulmonary fibrosis.

[0057] Certain novel compounds of the present invention show particularly high activity as EP4 receptor agonists.

[0058] The compounds of the present invention have been shown to have activity as EP4 receptor agonists. As shown by Caco-2 studies, the compounds of the present invention also have low gastrointestinal permeability. Therefore, it is believed that when administered orally, the compounds of the present invention exhibit low systemic bioavailability. Functional agonists of EP4 receptors expressed in the gastrointestinal tract have the potential to treat a range of gastrointestinal disorders. The combination of EP4 receptor agonist activity and low gastrointestinal permeability indicates that the compounds of the present invention can be used to treat a range of gastrointestinal disorders without cardiovascular side effects caused by systemic distribution.

[0059] In one aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0060]

[0061] in:

[0062] A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R', S(O)2R', S(O)2OR', SO2N(R')2, C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0063] Ring B is aryl or heteroaryl;

[0064] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0065] R 1 and R 2 Each independently is H, C 1-6 Alkyl, C 1-6 Alkoxy, or R 1 and R 2 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkane-1,1-diyl;

[0066] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0067] R 4 It is H, C 1-6 Alkyl, halogen, CN, NO2 or OR';

[0068] R 5 Is H or C 1-6 Alkylalkyl; or R 4 and R 5 Together with the pyrrolidine ring to which they are attached, they form C 1-6 Alkylene linker;

[0069] R 6 It is H, C 1-6 alkyl, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkoxy, halogen, CN, NO2, OR', COOR' or C(O)R',

[0070] R 7 Is OR', OC(O)R', OC(O)OR', CO2R', CON(R')2, SO2N(R')2, SO2R', OSO2R' or OSO2N(R')2;

[0071] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0072] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0073] n and m are each independently 0, 1, 2 or 3;

[0074] wherein at each occurrence, alkyl, alkylene and cycloalkyl are each optionally and independently substituted with OH', SH, CN, NO2, COOH, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0075] wherein heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO2, COOR', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0076] In some embodiments, the compound is a compound of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0077]

[0078] in:

[0079] A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R', S(O)2R', S(O)2OR', SO2N(R')2, C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0080] Ring B is aryl or heteroaryl;

[0081] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0082] R 1 and R 2 Each independently is H, C 1-6 Alkyl, C 1-6 Alkoxy, or R 1 and R 2 Together with the carbon atom to which they are attached, they form C 3-6 Cycloalkane-1,1-diyl;

[0083] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0084] R 4 It is C 1-6 Alkyl, halogen, CN, NO2 or OR';

[0085] R 5 Is H or C 1-6 Alkylalkyl; or R 4 and R 5 Together with the pyrrolidine ring to which they are attached, they form C 1-6Alkylene linker;

[0086] R 6 It is H, C 1-6 Alkyl, halogen, CN, NO2, OR', COOR' or C(O)R',

[0087] R 7 Is OR', OC(O)R', OC(O)OR', CO2R', CON(R')2, SO2N(R')2, SO2R', OSO2R' or OSO2N(R')2;

[0088] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0089] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0090] n and m are each independently 0, 1, 2 or 3;

[0091] wherein at each occurrence, alkyl, alkylene and cycloalkyl are each optionally and independently substituted with OH', SH, CN, NO2, COOH, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0092] wherein heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO2, COOR', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0093] In some embodiments, the compound is a compound of formula I, formula IIa, formula IIb, formula IIc, formula IId, formula IIe, formula (1), formula (1a), formula (1b), formula (2), formula (2a), formula (2b), formula 3, formula (3a), formula (3b), formula (4), formula (4a), formula (4b), formula (5), formula (5a), formula (5b), formula (6), formula (6a), formula (6b), formula (7), formula (7a), formula (7b), formula (8), formula (8a), formula (8b), formula (9), formula (9a), formula (9b), formula (10), formula (10a), formula (10b) or a pharmaceutically acceptable salt or tautomer thereof.

[0094] In some embodiments, the compound is a compound of formula I, formula IIa, formula IIb, formula IIc, formula IId, formula IIe, formula (1), formula (1a), formula (1b), formula (2), formula (2a), formula (2b), formula 3, formula (3a), formula (3b), formula (4), formula (4a), formula (4b), formula (5), formula (5a), formula (5b), formula (6), formula (6a), formula (6b), formula (7), formula (7a), formula (7b), formula (8), formula (8a), formula (8b), formula (9), formula (9a), formula (9b), formula (10), formula (10a), formula (10b) or a pharmaceutically acceptable salt thereof.

[0095] In some embodiments, A is CO2R', C(O)N(R')S(O)2R', S(O)2R', or heteroaryl.

[0096] In some embodiments, A is COOH or a 5-membered heteroaryl optionally substituted with OR' or SR'.

[0097] In some embodiments, Ring B is a 5- to 6-membered aryl or 5- to 6-membered heteroaryl, each of which is optionally and independently substituted by OR', SR', CN, NO2, CO2R', halogen, or C 1-4 Up to 3 instances of substitution in the alkyl group.

[0098] In some embodiments, Ring B is a 5- to 6-membered aryl or a 5- to 6-membered heteroaryl.

[0099] In some embodiments, Ring B is phenyl, which is optionally substituted with up to three instances of OH.

[0100] In some embodiments, Ring B is a 6-membered heteroaryl containing one or two nitrogen atoms, and each nitrogen is optionally substituted with oxo.

[0101] In some embodiments, Ring B is pyridine-N-oxide.

[0102] In some embodiments, each R 3 are independently selected from OR', halogen or C 1-6 alkyl.

[0103] In some embodiments, each R 3 is independently selected from OR' or alkyl; and n is 0 or 1. In some embodiments, n is 0.

[0104] In some embodiments, n is 1.

[0105] In some embodiments, n is 0 or 1.

[0106] In some embodiments, R 4is independently selected from OR' or halogen and; m is 0 or 1. In some embodiments, m is 0.

[0107] In some embodiments, m is 1.

[0108] In some embodiments, m is 0 or 1.

[0109] In some embodiments, R 5 It's H.

[0110] In some embodiments, R 4 and R 5 Together with the pyrrolidine ring to which they are attached they form a CH2 linker.

[0111] In some embodiments, R 4 With R 5 The carbon atom to which it is bonded is bonded to R 4 The attached pyrrolidine rings together form a C5 bridged bicyclic ring.

[0112] In some embodiments, R 4 and R 5 Together with the pyrrolidine ring to which they are attached they form a C5 bridged bicyclic ring.

[0113] In some embodiments, R 6 It is H, C 1-6 Alkyl, halogen or OR'.

[0114] In some embodiments, R 6 It is H, methyl or OH.

[0115] In some embodiments, R 7 It is OR', OC(O)R', CO2R', CON(R')2, SO2N(R')2, SO2R' or OSO2N(R')2.

[0116] In some embodiments, R 7 is OR', CO2R', CON(R')2, SO2N(R')2 or OSO2N(R')2. In some embodiments, R' is H.

[0117] In some embodiments, R 7 is OH, CO2H, CONH2, SO2NH2 or OSO2NH2. In some embodiments, R 7 It is CO2H, CONH2, SO2NH2 or OSO2NH2.

[0118] In some embodiments, R 7 It is CONH2.

[0119] In some embodiments, when R 7 When is OH, then U, V and Z are CH, and W is COH.

[0120] In some embodiments, provided herein are compounds of Formula IIa, Formula IIb, Formula IIc, Formula IId and Formula IIe, or pharmaceutically acceptable salts, solvates, hydrates, tautomers or optical isomers thereof:

[0121]

[0122] Among them, A, X, Y, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 As defined above.

[0123] In one embodiment, provided herein is a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0124]

[0125] wherein U, V, W and Z are each independently selected from CH, COH, N or N + -O - , and at least three of U, V, W and Z are CH.

[0126] In some embodiments, R 4 H, OH, F or R 5 Connect to form a CH2 bridge.

[0127] In some embodiments, R 5 H or R 4 Connect to form a CH2 bridge.

[0128] In some embodiments, A is selected from:

[0129]

[0130] Where R 9 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring.

[0131] In one embodiment, provided herein is a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0132]

[0133] in:

[0134] A is selected from:

[0135]

[0136] U, V, W and Z are each independently selected from CH, COH, N or N + -O - , and at least three of U, V, W and Z are CH;

[0137] X and Y are each independently selected from CH, CF, COH or N;

[0138] R 1 is H, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl, or R 1 With R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring;

[0139] R 2 is H, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl, or R 2 With R 1 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring;

[0140] R 3 is H, OH or F;

[0141] R 4 H, OH, F or R 5 ligated to form a CH2 linker;

[0142] R 5 H or R 4 connected to form a CH2 bridge;

[0143] R 6 is H, OH, CN, halogen, C optionally substituted by 1 to 3 fluorine atoms 1-3 alkoxy, or C optionally substituted by 1 to 3 fluorine atoms 1-3 alkyl;

[0144] R 7 is OH, CO2H, CONH2, SO2NH2 or OSO2NH2;

[0145] R 9 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring.

[0146] In some embodiments, provided herein are compounds of formula (1a) or (1b), or pharmaceutically acceptable salts, solvates, hydrates, tautomers or optical isomers thereof:

[0147]

[0148] Among them U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 As defined above.

[0149] In some embodiments, provided herein are compounds of formula (2), formula (2a) or formula (2b), or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0150]

[0151] Among them A, U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , R 6 and R 7 As defined above.

[0152] In some embodiments, the compound is a compound of formula (2a) or a pharmaceutically acceptable salt, solvate, hydrate or tautomer thereof:

[0153]

[0154] Among them U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , R 6 and R 7 As defined herein.

[0155] In some embodiments, the compound is a compound of formula (2a) or a pharmaceutically acceptable salt, solvate, hydrate or tautomer thereof:

[0156]

[0157] in:

[0158] U, V, W and Z are each independently selected from CH, COH, N or N + -O - , and at least three of U, V, W and Z are CH;

[0159] X and Y are each independently selected from CH, CF, COH or N;

[0160] R 1 is H, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl, or with R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring;

[0161] R 2 is H, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl, or R 2 With R 1 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring;

[0162] R 3 is H, OH or F;

[0163] R 4 H, OH, F or R 5 ligated to form a CH2 linker;

[0164] R 6 is H, OH, CN, halogen, C optionally substituted by 1 to 3 fluorine atoms 1-3 alkoxy, or C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl; and

[0165] R 7 It is OH, CO2H, CONH2, SO2NH2 or OSO2NH2.

[0166] In some embodiments, R 1 and R 2 are independently H, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkyl, or R 1 With R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring.

[0167] In some embodiments, R 1 is H or methyl, or R 1 With R 2 Connect to form a cyclopropane-1,1-diyl ring.

[0168] In some embodiments, provided herein are compounds of formula (3), formula (3a) or formula (3b), or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0169]

[0170] Among them A, U, V, W, X, Y, Z, R 3 , R 4 , R 6 and R 7 As defined above.

[0171] In some embodiments, the compound is a compound of formula (3a) or a pharmaceutically acceptable salt, solvate, hydrate or tautomer thereof:

[0172]

[0173] Among them U, V, W, X, Y, Z, R 3 , R 4 , R 6 and R 7 As defined herein.

[0174] In some embodiments, the compound is a compound of formula (3a) or a pharmaceutically acceptable salt, solvate, hydrate or tautomer thereof:

[0175]

[0176] in:

[0177] U, V, W and Z are each independently selected from CH, COH, N or N + -O - , and at least three of U, V, W and Z are CH;

[0178] X and Y are each independently selected from CH, CF, COH or N;

[0179] R 3 is H, OH or F;

[0180] R 4 H, OH, F or R 5 ligated to form a CH2 linker;

[0181] R 6 is H, OH, CN, halogen, C optionally substituted by 1 to 3 fluorine atoms 1-3 alkoxy, or C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl; and

[0182] R 7 It is OH, CO2H, CONH2, SO2NH2 or OSO2NH2.

[0183] In some embodiments, R 3 It is H, OH or F.

[0184] In some embodiments, R 3 It is H or OH.

[0185] In some embodiments, R 4 It is H, OH or F.

[0186] In some embodiments, R 6 is H, OH, CN, halogen, C optionally substituted by 1 to 3 fluorine atoms 1-3 alkoxy, or C optionally substituted by 1 to 3 fluorine atoms 1-3 alkyl.

[0187] In some embodiments, R 6 It is H, OH, CN or methyl.

[0188] In some embodiments, R 6 It is OH, CN or methyl.

[0189] In some embodiments, R 6 It's methyl.

[0190] In some embodiments, R 7 It is OH, CO2H, CONH2, SO2NH2 or OSO2NH2.

[0191] In some embodiments, R 7 It is CO2H, CONH2, SO2NH2 or OSO2NH2.

[0192] In some embodiments, R 7 It is CONH2 or SO2NH2.

[0193] In some embodiments, X and Y are each independently selected from CH, CF, COH, or N.

[0194] In some embodiments, provided herein are compounds of formula (4), formula (4a) or formula (4b), or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0195]

[0196] Among them, A, U, V, W, Z, R 3 , R 4 , R 6 and R 7 As defined above.

[0197] In some embodiments, provided herein are compounds of formula (5), formula (5a) or formula (5b), or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0198]

[0199] Among them, A, R 8 is H or OH, and R 3 , R 4 , R 6 and R 7 As defined above.

[0200] In some embodiments, provided herein are compounds of formula (6), formula (6a) or formula (6b), or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0201]

[0202] Among them, A, R 3 , R 6 , R 7 and R 8 As defined above.

[0203] In some embodiments, provided herein are compounds of formula (7), formula (7a) or formula (7b) or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0204]

[0205] Among them A, U, V, W, X, Y, Z, R 3 , R 6 and R 7 As defined above.

[0206] In some embodiments, provided herein are compounds of formula (8), formula (8a) or formula (8b) or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0207]

[0208] Among them A, U, V, W, X, Y, Z, R 3 , R 4 , R 6 and R 7 As defined above.

[0209] In some embodiments, provided herein are compounds of formula (9), formula (9a) or formula (9b) or pharmaceutically acceptable salts, solvates, hydrates, tautomers or optical isomers thereof:

[0210]

[0211] Among them A, U, V, W, X, Y, Z, R 1 , R 2, R 3 , R 6 and R 7 As defined herein.

[0212] In some embodiments, provided herein are compounds of formula (10), formula (10a) or formula (10b) or pharmaceutically acceptable salts, solvates, hydrates, tautomers or optical isomers thereof:

[0213]

[0214] Among them A, U, V, W, X, Y, Z, R 3 , R 6 and R 7 As defined herein.

[0215] In some embodiments, the compound is a compound of formula (1) or formula (9) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof,

[0216] in:

[0217] A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R', S(O)2R', S(O)2OR', SO2N(R')2, C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0218] U, V, W and Z are each independently selected from CH, COH, N or N + -O - , wherein at least three of U, V, W and Z are CH;

[0219] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0220] R 1 and R 2 Each independently is H, C 1-6 Alkyl, C 1-6 Alkoxy, or R 1 and R 2 Together with the carbon atom to which they are attached, they form C 3-6 Cycloalkane-1,1-diyl ring;

[0221] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0222] R 4 It is C 1-6Alkyl, halogen, CN, NO2 or OR';

[0223] R 5 Is H or C 1-6 alkyl;

[0224] R 6 It is H, C 1-6 Alkyl, halogen, CN, NO2, OR', CO2R' or C(O)R';

[0225] R 7 Is OR', OC(O)R', OC(O)OR', CO2R', CON(R')2, SO2N(R')2, SO2R', OSO2R' or OSO2N(R')2;

[0226] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0227] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0228] n and m are each independently 0, 1, 2 or 3;

[0229] wherein at each occurrence, alkyl, alkylene and cycloalkyl are each optionally and independently substituted by OH, SH, CN, NO2, COOH, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0230] wherein at each occurrence, heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO2, CO2R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0231] In some embodiments, the compound is a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof, wherein: A is COOH; U is CH; V is CH; W is CH, COH, N or N + -O - ; Z is CH or COH; X is CH; Y is CH or COH; R 1 is H or methyl, and R 2 Is H or R 1 With R 2 Connected to form a cyclopropane-1,1-diyl ring; R 3 is H or OH; R 4 is H or OH, and R 5 Is H or R 4 With R 5Connected to form a CH2 bridge; R 6 is H, OH, CN or methyl; and R 7 It is OH, CO2H, CONH2, SO2NH2 or OSO2NH2.

[0232] In some embodiments, the compound is a compound of formula (1) or formula (9), or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof, wherein: A is COOH; U is CH; V is CH; W is CH, COH, N or N + -O - ; Z is CH or COH; X is CH; Y is CH or COH; R 1 is H or methyl, and R 2 Is H or R 1 With R 2 Connected to form a cyclopropane-1,1-diyl ring; R 3 is H or OH; R 4 is H or OH; R 5 It is H; R 6 is H, OH, CN or methyl; and R 7 It is OH, CO2H, CONH2, SO2NH2 or OSO2NH2.

[0233] In some embodiments, A is selected from CO2H, tetrazole, 1,2,4- oxadiazole-5(2H)-one, 1,3,4- 2-oxadiazole-2(3H)-one, CONHSO2R 6 、CONHSO2Me、SO3H、1,3,4- oxadiazole-2(3H)-thione, 1,2,4- 1,2,4-thiadiazole-5(2H)-thione, 1,2,5-thiadiazolidine-3-one 1,1-dioxide and 2,4- Oxazolidinedione.

[0234] In some embodiments, A is selected from:

[0235]

[0236] In some embodiments, A is selected from CO2H and a tetrazole ring. A may be CO2H. A may be a tetrazole ring.

[0237] In some embodiments, U may be CH. U may be COH. U may be N. U may be N + -O - .

[0238] In some embodiments, U is CH.

[0239] In some embodiments, V may be CH. V may be COH. V may be N. V may be N + -O - .

[0240] In some embodiments, V is CH.

[0241] In some embodiments, W may be CH. W may be COH. W may be N. W may be N + -O - .

[0242] In some embodiments, W is CH, COH, N or N + -O - .

[0243] In some embodiments, W is CH or COH.

[0244] In some embodiments, Z may be CH. Z may be COH. Z may be N. Z may be N + -O - .

[0245] In some embodiments, Z is CH or COH.

[0246] In some embodiments, at least three of U, V, W, and Z are CH. U, V, and W may be CH. U, V, and Z may be CH. U, W, and Z may be CH. V, W, and Z may be CH. U, V, W, and Z may be CH. U, V, and Z may be CH and W may be COH.

[0247] In some embodiments, X may be CH. X may be CF. X may be COH. X may be N.

[0248] In some embodiments, Y may be CH. Y may be CF. Y may be COH. Y may be N.

[0249] In some embodiments, Y can be CH or COH.

[0250] In the compounds herein, at least one of X and Y is CH. Both X and Y may be CH.

[0251] In some embodiments, X may be CH, and Y may be CH or COH.

[0252] In some embodiments, R 1 It may be H or C optionally substituted by 1 to 3 fluorine atoms. 1-3 Alkyl.1 It can be H. R 1 It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl. 1 Can be C 1-3 Alkyl. 1 Can be used with R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring. 1 Can be used with R 2 Connect to form C 3-6 Cycloalkyl ring. 1 It can be H or methyl, or R 1 Can be used with R 2 Connect to form a cyclopropane ring. 1 It may be a methyl group optionally substituted by 1 to 3 fluorine atoms. 1 It can be methyl. 1 Can be used with R 2 are connected to form a cyclopropane ring optionally substituted with 1 to 3 fluorine atoms. 1 Can be used with R 2 Connect to form a cyclopropane ring.

[0253] In some embodiments, R 2 It can be H. R 2 It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl. 2 Can be C 1-3 Alkyl. 2 Can be used with R 1 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring. 2 Can be used with R 1 Connect to form C 3-6 Cycloalkyl ring. 2 Can be used with R 1 are connected to form a cyclopropane ring optionally substituted with 1 to 3 fluorine atoms. 2 Can be used with R 1 Connect to form a cyclopropane ring.

[0254] In some embodiments, R 3 It can be H. R 3 It can be OH. 3 It can be F.

[0255] In some embodiments, R 4 It can be H. R 4 It can be OH. 4 It can be F. R 4 Can be used with R5 Connect to form a CH2 bridge.

[0256] In some embodiments, R 5 It can be H. R 5 Can be used with R 4 Connect to form a CH2 bridge.

[0257] In some embodiments, R 6 and R 7 Together with the rings they are connected to, they can be selected from:

[0258]

[0259] In some embodiments, R 6 It can be H, OH, CN or methyl. 6 It can be H. R 6 It can be OH. 6 Can be CN. 6 It can be halogen. 6 It can be F. R 6 It can be Cl. 6 It can be Br. 6 It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkoxy. 6 Can be C 1-3 Alkoxy. 6 It may be a methoxy group optionally substituted by 1 to 3 fluorine atoms. 6 It can be methoxy. 6 It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl. 6 Can be C 1-3 Alkyl. 6 It can be H or methyl. 6 It may be a methyl group optionally substituted by 1 to 3 fluorine atoms. 6 It may be methyl.

[0260] In some embodiments, R 7 It can be OH. 7 It can be CO2H. 7 It can be CONH2. 7 It can be SO2NH2. 7 It can be OSO2NH2.

[0261] In some embodiments, R 8 It can be H. R 8 It can be OH.

[0262] In some embodiments, R9 Can be C 1-3 Alkyl. 9 Can be C 3-6 Cycloalkyl ring. 9 It may be methyl.

[0263] In some embodiments, A may be CO2H; U, V and Z may be CH; W may be COH; R 1 It can be methyl; R 2 can be H; and R 6 It may be methyl.

[0264] In some embodiments, the compounds or pharmaceutically acceptable salts disclosed herein have EP4 receptor agonist activity.

[0265] In some embodiments, the present invention includes a pharmaceutical composition comprising a compound, a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer described herein and a pharmaceutically acceptable excipient.

[0266] In some embodiments, the pharmaceutical composition further comprises at least one additional therapeutic agent selected from the group consisting of aminosalicylates, corticosteroids, immunomodulators, and combinations thereof.

[0267] In some embodiments, disclosed herein are methods of treating a disease mediated by an EP4 receptor, the method comprising administering to a patient in need thereof a compound or a pharmaceutically acceptable salt disclosed herein.

[0268] In some embodiments, the method of treating an EP4 receptor mediated disease is a gastrointestinal disorder.

[0269] In some embodiments, the gastrointestinal disorder is selected from constipation disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergies or food intolerances, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced bowel disease, NSAID-induced gastric and intestinal damage, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrheal diseases, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis, or pulmonary diseases and disorders, such as chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperresponsive airway disorders, and idiopathic pulmonary fibrosis.

[0270] In some embodiments, the compound of Formula I or Formula (1) is a compound listed in Table 1 or a pharmaceutically acceptable salt, solvate, hydrate or tautomer thereof.

[0271] In some embodiments, the compound of Formula I or Formula (1) is a compound listed in Table 1 or a pharmaceutically acceptable salt thereof.

[0272] Table 1: Exemplary compounds of formula (I)

[0273]

[0274]

[0275]

[0276]

[0277] abbreviation

[0278] aq aqueous solution

[0279] Bn Benzyl

[0280] DCM Dichloromethane

[0281] DMA Dimethylacetamide

[0282] DMF Dimethylformamide

[0283] dppf 1,1'-Bis(diphenylphosphino)ferrocene

[0284] EDCI 1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide

[0285] FA Formic acid

[0286] EtOAc Ethyl acetate

[0287] HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-Oxide Hexafluorophosphate

[0288] HOBt Hydroxybenzotriazole

[0289] HPLC High Performance Liquid Chromatography

[0290] hr hours

[0291] hrs hours

[0292] LC / MS Liquid chromatography mass spectrometry

[0293] M Moore

[0294] MeCN Acetonitrile

[0295] MeOH Methanol

[0296] N equivalent (normal)

[0297] Preparative HPLC Preparative High Performance Liquid Chromatography

[0298] RT Room temperature

[0299] sat saturated

[0300] THF Tetrahydrofuran

[0301] UPLC Ultra-Performance Liquid Chromatography

[0302] definition

[0303] In this application, unless otherwise indicated, the following definitions apply.

[0304] The term "treatment" in connection with the use of any of the compounds described herein, including those of Formula I, IIa, IIb, IIc, IId, IIe, (1), (1a), (1b), (2), (2a), (2b), (3), (3a), (3b), (4), (4a), (4b), (5), (5a), (5b), (6), (6a), (6b), (7), (7a), (7b), (8), (8a), (8b), (9), (9a), (9b), (10), (10a), and (10b) is used to describe any form of intervention in which the compound is administered to a subject having, at risk of, or potentially at risk of the disease or condition in question. Thus, the term "treatment" includes both prophylactic (preventive) treatment and treatment that results in measurable or detectable symptoms of the disease or condition.

[0305] The term "therapeutically effective amount" (e.g., in relation to a method of treating a disease or condition) refers to an amount of a compound that is effective to produce the desired therapeutic effect. For example, if the condition is pain, the therapeutically effective amount is an amount sufficient to provide a desired level of pain relief. The desired level of pain relief can be, for example, complete elimination of pain or a reduction in the severity of pain.

[0306] As used herein, the term "hydroxyl" or "hydroxy" refers to an -OH moiety.

[0307] As used herein, "alkyl" refers to a saturated aliphatic hydrocarbon group containing 1 to 12 (e.g., 1 to 8, 1 to 6, or 1 to 4) carbon atoms. The alkyl group can be straight chain or branched. Some examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-heptyl, or 2-ethylhexyl. The alkyl group may be substituted (i.e., optionally substituted) with one or more substituents, such as halogen, phosphate, alicyclic [e.g., cycloalkyl or cycloalkenyl], heteroalicyclic [e.g., heterocycloalkyl or heterocycloalkenyl], aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl [e.g., (aliphatic)carbonyl, (alicyclic)carbonyl, or (heteroalicyclic)carbonyl], nitro, cyano, amide [e.g., (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heterocycloalkylalkyl)carbonylamino, [e.g., aliphatic amino, alicyclic amino, or heteroalicyclic amino], sulfonyl [e.g., aliphatic -SO2-], sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfonamide, oxo, carboxyl, carbamoyl, alicyclic oxy, heteroalicyclic oxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, or hydroxyl. Without limitation, some examples of substituted alkyl groups include carboxyalkyl (e.g., HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, aralkyl, (alkoxyaryl)alkyl, (sulfonylamino)alkyl (e.g., (alkyl-SO2-amino)alkyl), aminoalkyl, amidoalkyl, (alicyclic)alkyl, or haloalkyl.

[0308] As used herein, an "alkylene" group refers to a divalent branched alkyl or divalent straight chain alkyl containing 2 to 12 (e.g., 2 to 8, 2 to 6, or 2 to 4) carbon atoms and is used to connect two chemical moieties. Some examples of alkylene include, but are not limited to, methylene, ethylene, propylene, butylene, isopropylidene (methylethylene) and isobutylene (2-methylpropylene). Alkylene can be substituted (i.e., optionally substituted) by one or more substituents as defined in alkyl.

[0309] As used herein, "amido" encompasses both "aminocarbonyl" and "carbonylamino". These terms, when used alone or in conjunction with another group, refer to an amide group, for example, when used terminally, the term refers to -N(R X )-C(O)-R Y or -C(O)-N(R X)2, and when used internally, the term refers to -C(O)-N(R X )-or-N(R X )-C(O)-, where R X and R Y It can be aliphatic, alicyclic, aryl, araliphatic, heteroalicyclic, heteroaryl or heteroaraliphatic. Some examples of amide groups include alkylamide groups (e.g., alkylcarbonylamino or alkylaminocarbonyl), (heteroalicyclic)amide groups, (heteroaralkyl)amide groups, (heteroaryl)amide groups, (heterocycloalkyl)alkylamide groups, arylamide groups, aralkylamide groups, (cycloalkyl)alkylamide groups or cycloalkylamide groups.

[0310] As used herein, "amino" refers to -NR X R Y , where R X and R Y Each of is independently hydrogen, aliphatic, alicyclic, (alicyclic) aliphatic, aryl, aromatic aliphatic, heteroalicyclic, (heteroalicyclic) aliphatic, heteroaryl, carboxyl, sulfanyl, sulfinyl, sulfonyl, (aliphatic) carbonyl, (alicyclic) carbonyl, ((alicyclic) aliphatic) carbonyl, arylcarbonyl, (aromatic aliphatic) carbonyl, (heteroalicyclic) carbonyl, ((heteroalicyclic) aliphatic) carbonyl, (heteroaryl) carbonyl or (heteroaromatic aliphatic) carbonyl, each of which is defined herein and is optionally substituted. Some examples of amino include alkylamino, dialkylamino or arylamino. When the term "amino" is not a terminal group (e.g., alkylcarbonylamino), it is represented by -NR X - indicates that R X Has the same meaning as defined above.

[0311] As used herein, "aryl" used alone or as part of a larger moiety in "aralkyl", "aralkyloxy" or "aryloxyalkyl" refers to monocyclic (e.g., phenyl); bicyclic (e.g., indenyl, naphthyl, tetrahydronaphthyl, tetrahydroindenyl); and tricyclic (e.g., fluorenyl, tetrahydrofluorenyl, or tetrahydroanthracenyl, anthracenyl) ring systems, wherein the monocyclic system is aromatic or at least one of the rings in the bicyclic or tricyclic system is aromatic. Bicyclic and tricyclic groups include benzo-fused 2- to 3-membered carbocyclic rings. For example, benzo-fused groups include benzo-fused 2- to 3-membered carbocyclic rings with two or more C 4-8The aryl group is optionally substituted with one or more substituents including: aliphatic [e.g., alkyl, alkenyl, or alkynyl]; alicyclic; (alicyclic) aliphatic; heteroalicyclic; (heteroalicyclic) aliphatic; aryl; heteroaryl; alkoxy; (alicyclic) oxy; (heteroalicyclic) oxy; aryloxy; heteroaryloxy; (aroliphatic) oxy; (heteroaroaliphatic) oxy; aroyl; heteroaroyl; amino; oxo (on the non-aromatic carbocyclic ring of the benzo-fused bicyclic or tricyclic aryl group); nitro; carboxyl; amide; acyl [e.g., For example, (aliphatic) carbonyl; (alicyclic) carbonyl; ((alicyclic) aliphatic) carbonyl; (aromatic) carbonyl; (heteroalicyclic) carbonyl; ((heteroalicyclic) aliphatic) carbonyl; or (heteroaromatic) carbonyl]; sulfonyl [e.g., aliphatic -SO2- or amino-SO2-]; sulfinyl [e.g., aliphatic -S(O)- or alicyclic -S(O)-]; sulfanyl [e.g., aliphatic -S-]; cyano; halogen; hydroxyl; mercapto; sulfoxyl; urea; thiourea; sulfamoyl; sulfonamide; or carbamoyl. Alternatively, the aryl group can be unsubstituted.

[0312] Some non-limiting examples of substituted aryl groups include haloaryl [e.g., monohaloaryl, dihaloaryl (e.g., p-, m-dihaloaryl), and (trihalo)aryl]; (carboxy)aryl [e.g., (alkoxycarbonyl)aryl, ((aralkyl)carbonyloxy)aryl, and (alkoxycarbonyl)aryl]; (amido)aryl [e.g., (aminocarbonyl)aryl, (((alkylamino)alkyl)aminocarbonyl)aryl, (alkylcarbonyl)aminoaryl, (arylaminocarbonyl)aryl, and (((heteroaryl)amino)carbonyl)aryl]; aminoaryl [e.g., ((alkylsulfonyl)amino)aryl or ((dialkyl)amino)aryl]; (cyanoalkyl)aryl; (alkoxy)aryl; (sulfamoyl)aryl )aryl [e.g., (aminosulfonyl)aryl]; (alkylsulfonyl)aryl; (cyano)aryl; (hydroxyalkyl)aryl; ((alkoxy)alkyl)aryl; (hydroxy)aryl, ((carboxy)alkyl)aryl; (((dialkyl)amino)alkyl)aryl; (nitroalkyl)aryl; (((alkylsulfonyl)amino)alkyl)aryl; ((heteroalicyclic)carbonyl)aryl; ((alkylsulfonyl)alkyl)aryl; (cyanoalkyl)aryl; (hydroxyalkyl)aryl; (alkylcarbonyl)aryl; alkylaryl; (trihaloalkyl)aryl; p-amino-m-alkoxycarbonylaryl; p-amino-m-cyanoaryl; p-halo-m-aminoaryl; or (m-(heteroalicyclic)-o-(alkyl))aryl.

[0313] As used herein, a "cycloalkyl" group refers to a saturated carbocyclic monocyclic or bicyclic (fused or bridged) ring of 3 to 10 (e.g., 5 to 10) carbon atoms. Some examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, norbornyl, cubicyl, octahydro-indenyl, decahydro-naphthyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, bicyclo[3.3.2.]decyl, bicyclo[2.2.2]octyl, adamantyl, or ((aminocarbonyl)cycloalkyl)cycloalkyl.

[0314] The cycloalkyl radical may be optionally substituted with one or more substituents such as phosphate, aliphatic [e.g., alkyl, alkenyl, or alkynyl], cycloaliphatic, (cycloaliphatic)aliphatic, heterocycloaliphatic, (heterocycloaliphatic)aliphatic, aryl, heteroaryl, alkoxy, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy, aryloxy, heteroaryloxy, (araliphatic)oxy, (heteroaroaliphatic)oxy, aroyl, heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino, (cycloaliphatic)carbonylamino, ((cycloaliphatic)aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino, (heterocycloaliphatic)carbonylamino, ((heterocycloaliphatic)aliphatic) [e.g., alkyl-S-], sulfoxyl, urea, thiourea, sulfamoyl, sulfonyl, or carbamoyl groups.

[0315] As used herein, "heterocycloalkyl" refers to a 3- to 10-membered monocyclic or bicyclic (fused or bridged) (e.g., a 5- to 10-membered monocyclic or bicyclic) saturated ring structure in which one or more of the ring atoms is a heteroatom (e.g., N, O, S, or a combination thereof). Some examples of heterocycloalkyl include piperidinyl, piperazinyl, tetrahydropyranyl, tetrahydrofuranyl, 1,4-dioxolanyl, 1,4-dithianyl, 1,3-dioxolanyl, Oxazoline, iso oxazolinyl, morpholinyl, thiomorpholyl, octahydrobenzofuranyl, octahydrochromenyl, octahydrothiochromenyl, octahydroindolyl, octahydropyridinyl, decahydroquinolinyl, octahydrobenzo[b]thienyl, 2-oxa-bicyclo[2.2.2]octyl, 1-aza-bicyclo[2.2.2]octyl, 3-aza-bicyclo[3.2.1]octyl and 2,6-dioxa-tricyclo[3.3.1.0 3,7 A monocyclic heterocycloalkyl group may be fused with a phenyl moiety to form a structure such as tetrahydroisoquinoline, which would be classified as a heteroaryl group.

[0316] The heterocycloalkyl radical may be optionally substituted with one or more substituents such as phosphate, aliphatic [e.g., alkyl, alkenyl, or alkynyl], alicyclic, (alicyclic)aliphatic, heteroalicyclic, (heteroalicyclic)aliphatic, aryl, heteroaryl, alkoxy, (alicyclic)oxy, (heteroalicyclic)oxy, aryloxy, heteroaryloxy, (araliphatic)oxy, (heteroaraliphatic)oxy, aroyl, heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino, (alicyclic)carbonylamino, ((alicyclic)aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino, (heteroalicyclic)carbonylamino, ((heteroalicyclic) [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl, ((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaroaliphatic)carbonyl], nitro, cyano, halogen, hydroxy, mercapto, sulfonyl [e.g., alkylsulfonyl or arylsulfonyl], sulfinyl [e.g., alkylsulfinyl], sulfanyl [e.g., alkylsulfanyl], sulfoxy, urea, thiourea, sulfamoyl, sulfonamide, oxo, or carbamoyl.

[0317] As used herein, "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic ring system having 4 to 15 ring atoms, wherein one or more of the ring atoms is a heteroatom (e.g., N, O, S, or a combination thereof), and wherein the monocyclic system is aromatic, or at least one ring of the bicyclic or tricyclic system is aromatic.

[0318] Heteroaryl includes benzo-fused ring systems having 2 to 3 rings. For example, benzo-fused groups include benzo fused to one or two 4 to 8-membered heteroalicyclic moieties (e.g., indolizyl, indolyl, isoindolyl, 3H-indolyl, dihydroindole, benzo[b]furanyl, benzo[b]thienyl, quinolyl, or isoquinolyl). Some examples of heteroaryl are azetidinyl, pyridyl, 1H-indazolyl, furanyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuranyl, isoquinolyl, benzothiazolyl, tonyl, thioxanthenyl, phenothiazinyl, indoline, benzo[1,3]dioxole, benzo[b]furanyl, benzo[b]thienyl, indazolyl, benzimidazolyl, benzothiazolyl, puryl, cinnolyl, quinolyl, quinazolinyl, cinnolinyl, phthalazyl, quinazolinyl, quinoxalyl, isoquinolyl, 4H-quinolizyl, benzo-1,2,5-thiadiazolyl or 1,8-naphthyridyl.

[0319] Heteroaryl groups include N-oxide heteroaryl compounds which are oxidized at the nitrogen atom, for example pyridine, pyrimidine, pyrazine, pyrrole, imidazole, thiazole, quinoline or isoquinoline. Examples of N-oxide heteroaryl groups are those having the formula C5H5N - →O + Pyridine-N-oxide.

[0320] Without limitation, monocyclic heteroaryl groups include furanyl, thiophene-yl, 2H-pyrrolyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isocyanate oxazolyl, isothiazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4-H-pyranyl, pyridinyl, pyridazyl, pyrimidinyl, pyrazolyl, pyrazyl or 1,3,5-triazyl. Monocyclic heteroaryl groups are numbered according to standard chemical nomenclature.

[0321] Without limitation, bicyclic heteroaryl groups include indolizyl, indolyl, isoindolyl, 3H-indolyl, dihydroindole, benzo[b]furanyl, benzo[b]thienyl, quinolyl, isoquinolyl, indolizyl, isoindolyl, indolyl, benzo[b]furanyl, bexo[b]thienyl, indazolyl, benzimidazolyl, benzothiazolyl, purinyl, 4H-quinolizinyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl or pteridinyl. Bicyclic heteroaryl groups are numbered according to standard chemical nomenclature.

[0322] The heteroaryl group is optionally substituted by one or more substituents such as: aliphatic [e.g., alkyl, alkenyl, or alkynyl]; alicyclic; (alicyclic)aliphatic; heteroalicyclic; (heteroalicyclic)aliphatic; aryl; heteroaryl; alkoxy; (alicyclic)oxy; (heteroalicyclic)oxy; aryloxy; heteroaryloxy; (aroliphatic)oxy; (heteroaroaliphatic)oxy; aroyl; heteroaroyl; amino; oxo (on the non-aromatic carbocyclic or heterocyclic ring of the bicyclic or tricyclic heteroaryl); carboxyl; amide The heteroaryl group may be substituted with acyl, acyl [e.g., aliphatic carbonyl; (alicyclic) carbonyl; ((alicyclic) aliphatic) carbonyl; (aromatic) carbonyl; (heteroalicyclic) carbonyl; ((heteroalicyclic) aliphatic) carbonyl; or (heteroaromatic) carbonyl]; sulfonyl [e.g., aliphatic sulfonyl or aminosulfonyl]; sulfinyl [e.g., aliphatic sulfinyl]; sulfanyl [e.g., aliphatic sulfanyl]; nitro; cyano; halogen; hydroxyl; mercapto; sulfoxyl; urea; thiourea; sulfamoyl; sulfonamide; or carbamoyl. Alternatively, the heteroaryl group may be unsubstituted.

[0323] Some non-limiting examples of substituted heteroaryl groups include (halo)heteroaryl [e.g., monohaloheteroaryl and dihaloheteroaryl]; (carboxy)heteroaryl [e.g., (alkoxycarbonyl)heteroaryl]; cyanoheteroaryl; aminoheteroaryl [e.g., ((alkylsulfonyl)amino)heteroaryl and ((dialkyl)amino)heteroaryl]; (amido)heteroaryl [e.g., aminocarbonylheteroaryl, ((alkylcarbonyl)amino)heteroaryl, ((((alkyl)amino)alkyl)aminocarbonyl)heteroaryl, (((heteroaryl)amino)carbonyl)heteroaryl, ((heteroalicyclic)carbonyl)heteroaryl, and ((alkylcarbonyl)amino)heteroaryl]; (cyanoalkyl)heteroaryl; (alkoxy)heteroaryl; ((alkylsulfonyl)amino)heteroaryl; ((alkylsulfonyl)amino)heteroaryl; ((dialkyl)amino)heteroaryl) heteroaryl]; (alkyl)heteroaryl ...

[0324] As used herein, an "alkoxy" group refers to an alkyl-O- group, wherein "alkyl" has been previously defined.

[0325] As used herein, a "carboxyl" group, when used as a terminal group, refers to -COOH, -COOR X 、-OC(O)H、-OC(O)R X; or when used as an internal group, a "carboxy" group refers to -OC(O)- or -C(O)O-.

[0326] As used herein, a "thiol" group refers to -SH.

[0327] As used herein, a "sulfo" group when used terminally refers to -SO3H or -SO3R X , or -S(O)3- when used internally.

[0328] As used herein, a "sulfonamide" group when used terminally refers to a group of the structure -NR X -S(O)2-NR Y R Z , and when used internally refers to the structure -NR X -S(O)2-NR Y -, where R X , R Y and R Z Defined above.

[0329] As used herein, a "sulfamoyl" group refers to a group of the structure -OS(O)2-NR Y R Z , where R Y and R Z Defined above.

[0330] As used herein, a "sulfonamide" group when used terminally refers to the structure -S(O)2-NR x R y or -NR x -S(O)2-R z ; or when used internally, -S(O)2-NR x -or-NR x -S(O)2-, where R x , R y and R Z Defined above.

[0331] As used herein, a "sulfanyl" group when used terminally refers to -SR X , and when used internally refers to -S-, where R X Some examples of sulfanyl groups include aliphatic-S-, alicyclic-S-, aryl-S-, and the like.

[0332] As used herein, a "halogen" or "halo" group refers to fluorine, chlorine, bromine, or iodine.

[0333] As used herein, "oxo" refers to =0.

[0334] As used herein, the term "vicinal" generally refers to the positioning of substituents on a group containing two or more carbon atoms wherein the substituents are attached to adjacent carbon atoms.

[0335] As used herein, the term "geminal" generally refers to the position of substituents on a group containing two or more carbon atoms, wherein the substituents are attached to the same carbon atom.

[0336] The terms "terminal" and "internal" refer to the position of a group within a substituent. A group is terminal when it is present at the end of a substituent and is no longer bonded to the rest of the chemical structure. X O(O)C-alkyl) is an example of a carboxyl group used terminally. When the group is present in the middle of a substituent in a chemical structure, the group is internal. Alkylcarboxyl (e.g., alkyl-C(O)O- or alkyl-OC(O)-) and alkylcarboxyaryl (e.g., alkyl-C(O)O-aryl or alkyl-O(CO)-aryl-) are some examples of carboxyl groups used internally.

[0337] As used herein, "aliphatic chain" refers to a branched or straight chain aliphatic group (e.g., an alkyl, alkenyl, or alkynyl group). A straight chain aliphatic chain has the structure -[CH2] v -, where v is 1 to 12. A branched aliphatic chain is a straight aliphatic chain substituted with one or more aliphatic groups. A branched aliphatic chain has the structure -[CQQ] v -, wherein Q is independently hydrogen or an aliphatic group; however, in at least one instance, Q shall be an aliphatic group. The term aliphatic chain includes alkyl chains, alkenyl chains, and alkynyl chains, wherein alkyl, alkenyl, and alkynyl are defined above.

[0338] The phrase "optionally substituted" is used interchangeably herein with the phrase "substituted or unsubstituted". As described herein, the compounds of the invention may be optionally substituted with one or more substituents, such as are generally exemplified above, or exemplified by specific classes, subclasses, and species of the invention. Unless otherwise indicated, each specific group of variables described herein may be optionally substituted with one or more substituents described herein. Each substituent of a specific group is also optionally substituted with one to three of halogen, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, alicyclic, heteroalicyclic, heteroaryl, haloalkyl, and alkyl. For example, an alkyl group may be substituted with an alkylsulfanyl group, and an alkylsulfanyl group may be optionally substituted with one to three of halogen, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, haloalkyl, and alkyl. As another example, the cycloalkyl moiety of (cycloalkyl)carbonylamino may be optionally substituted with one to three of halogen, cyano, alkoxy, hydroxy, nitro, haloalkyl and alkyl. When two alkoxy groups are bound to the same atom or adjacent atoms, the two alkoxy groups may form a ring together with the atoms to which they are bound.

[0339] As used herein, the term "substituted", whether or not there is the term "optionally" in front, generally refers to that the hydrogen atoms in a given structure are replaced by the group of a specific substituent. Specific substituents are described in the above definition and in the following description of compounds and embodiments thereof. Unless otherwise indicated, the optionally substituted group may have a substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted by more than one substituent selected from a specific group, the substituent at each position may be the same or different. Ring substituents such as heterocycloalkyl can be combined with another ring such as cycloalkyl to form a spiro-bicyclic system, such as two rings share a common atom. As will be appreciated by those of ordinary skill in the art, the combination of substituents contemplated by the present invention is those combinations that result in the formation of stable or chemically feasible compounds.

[0340] As used herein, the phrase "stable or chemically feasible" refers to a compound that is not substantially altered when subjected to conditions that allow its production, detection, and preferably its recovery, purification, and use for one or more purposes disclosed herein. In some embodiments, a stable compound or a chemically feasible compound is one that is not substantially altered when kept at 40° C. or less for at least one week in the absence of moisture or other chemically reactive conditions.

[0341] To the extent that any of the compounds described have chiral centers, the present invention extends to all optical isomers of such compounds, whether in racemic or resolved enantiomeric form. The invention described herein relates to all crystalline forms, solvates and hydrates of any disclosed compound, no matter how prepared. To the extent that any of the compounds disclosed herein have acid or basic centers such as carboxylate / ester or amino groups, then all salt forms of the compounds are included herein. In the case of pharmaceutical use, the salts should be considered as pharmaceutically acceptable salts.

[0342] Salts or pharmaceutically acceptable salts that may be mentioned include acid addition salts and base addition salts. Such salts can be formed, for example, by the following conventional methods: by reacting the compound in free acid or free base form with one or more equivalents of a suitable acid or base, optionally in a solvent or medium in which the salt is not soluble, followed by removal of the solvent or medium using standard techniques (e.g. in vacuo, by freeze drying or by filtration). Salts can also be prepared by exchanging the counterion of the compound in salt form with another counterion, for example using a suitable ion exchange resin.

[0343] Some examples of pharmaceutically acceptable salts include acid addition salts derived from inorganic and organic acids, and salts derived from metals such as sodium, magnesium, potassium, and calcium.

[0344] Some examples of acid addition salts include acid addition salts formed with acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, arylsulfonic acids (e.g., benzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, and p-toluenesulfonic acid), ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzoic acid, 4-acetamidobenzoic acid, butyric acid, (+) camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acids (e.g., D-gluconic acid), glucuronic acids (e.g.,

[0013] Examples of the present invention include, but are not limited to, D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-oxoglutaric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, isethionic acid, lactic acid (e.g., (+)-L-lactic acid and (±)-DL-lactic acid), lactobionic acid, maleic acid, malic acid (e.g., (-)-L-malic acid), malonic acid, (±)-DL-mandelic acid, metaphosphoric acid, methanesulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, tartaric acid (e.g., (+)-L-tartaric acid), thiocyanic acid, undecylenic acid, and valeric acid.

[0345] Any solvate of the compound and its salt is also contemplated.Preferred solvates are solvates formed by incorporating nontoxic pharmaceutically acceptable solvent molecules (hereinafter referred to as solvating solvents) into the solid state structure (e.g., crystal structure) of the compound of the present invention.Some examples of such solvents include water, alcohols (e.g., ethanol, isopropanol and butanol) and dimethyl sulfoxide.Solvates can be prepared by recrystallizing the compound of the present invention with a solvent or a mixture of solvents containing a solvating solvent. Whether a solvate has been formed in any given case can be determined by analyzing the crystals of the compound using known and standard techniques such as thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and X-ray crystallography.

[0346] Solvates can be stoichiometric or non-stoichiometric solvates. Specific solvates can be hydrates, and some examples of hydrates include hemihydrates, monohydrates and dihydrates. For a more detailed discussion of solvates and methods for preparing and characterizing them, see Bryn et al, Solid-State Chemistry of Drugs, Second Edition, published by SSCI, Inc of West Lafayette, IN, USA, 1999, ISBN 0-967-06710-3.

[0347] The term "pharmaceutical composition" in the context of the present invention means a composition comprising an active agent and additionally comprising one or more pharmaceutically acceptable carriers or pharmaceutically acceptable excipients. The composition may also comprise ingredients selected from, for example, diluents, adjuvants, excipients, carriers, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatics, antibacterial agents, antifungal agents, lubricants and dispersants, depending on the nature of the administration mode and dosage form. The composition may take the form of, for example, tablets, dragees, powders, elixirs, syrups, liquid preparations (including suspensions, sprays, inhalants, tablets, lozenges, emulsions, solutions, cachets, granules, capsules and suppositories) and liquid preparations for injection (including liposomal preparations).

[0348] The compounds of the present invention may contain one or more isotopic substitutions, and reference to a particular element includes within its scope all isotopes of that element. For example, reference to hydrogen includes within its scope all isotopes of that element. 1 H. 2 H(D) and 3 H(T). Similarly, references to carbon and oxygen include within their scope 12 C. 13 C and14 C and 16 O and 18 O. In a similar manner, references to specific functional groups also include isotopic variations within their scope unless the context indicates otherwise. For example, references to alkyl groups such as ethyl or alkoxy groups (e.g., methoxy) also include variants in which one or more hydrogen atoms in the group are in deuterated or tritium isotopic form, such as ethyl in which all five hydrogen atoms are in deuterated isotopic form (perdeuterated ethyl) or methoxy in which all three hydrogen atoms are in deuterated isotopic form (trideuterated methoxy). Isotopes may be radioactive or non-radioactive.

[0349] The therapeutic dose may vary depending on the patient's needs, the severity of the condition being treated, and the compound being employed. It is within the skill of the art to determine the appropriate dose for a particular situation. Typically, treatment is initiated with a smaller dose that is less than the optimal dose of the compound. Thereafter, the dose is increased in small increments until the optimal effect is achieved in the situation. For convenience, the total daily dose may be divided and administered in batches throughout the day if desired.

[0350] Of course, the size of the compound effective dose will vary with the severity of the condition to be treated and the specific compound and its route of administration. The selection of appropriate doses is within the capabilities of those of ordinary skill in the art, without excessive burden. Typically, daily dose ranges can be from about 10 μg to about 30 mg / kg human and non-human animal body weight, preferably from about 50 μg to about 30 mg / kg human and non-human animal body weight, for example, from about 50 μg to about 10 mg / kg human and non-human animal body weight, for example, from about 100 μg to about 30 mg / kg human and non-human animal body weight, for example, from about 100 μg to about 10 mg / kg human and non-human animal body weight, and most preferably from about 100 μg to about 1 mg / kg human and non-human animal body weight.

[0351] Combination therapy

[0352] In the methods or pharmaceutical compositions of the present invention, an effective amount can be achieved by using a compound of the present invention (including a pharmaceutically acceptable salt or solvate (e.g., hydrate)) alone or in combination with another suitable therapeutic agent (e.g., an antiviral agent or a vaccine). When "combination therapy" is employed, an effective amount can be achieved by using a first amount of a compound of the present invention and a second amount of another suitable therapeutic agent.

[0353] In another embodiment of the present invention, the compounds of the present invention and the additional therapeutic agent are each administered in an effective amount (i.e., if administered alone, each is administered in an amount that would be therapeutically effective). In another embodiment, the compounds of the present invention and the additional therapeutic agent are each administered in an amount that alone does not provide a therapeutic effect (a subtherapeutic dose). In another embodiment, the compounds of the present invention may be administered in an effective amount, while the additional therapeutic agent is administered in a subtherapeutic dose. In another embodiment, the compounds of the present invention may be administered in a subtherapeutic dose, while the additional therapeutic agent, such as a suitable cancer therapeutic agent, is administered in an effective amount.

[0354] As used herein, the terms "combination" or "co-administration" are used interchangeably to refer to the use of more than one treatment (e.g., one or more prophylactic and / or therapeutic agents). The use of these terms does not limit the order in which the treatments (e.g., prophylactic and / or therapeutic agents) are administered to a subject.

[0355] Co-administration encompasses administration of the co-administered first and second amounts of the compound in a substantially simultaneous manner, such as in a single pharmaceutical composition, e.g., a capsule or tablet having a fixed ratio of the first and second amounts, or in a plurality of separate capsules or tablets of each. In addition, such co-administration also encompasses the use of each compound in a sequential manner in either order.

[0356] In one embodiment, the compound of the invention and the additional therapeutic agent are administered to a subject separately, sequentially or simultaneously.

[0357] When co-administration involves separate administration of a first amount of a compound of the invention and a second amount of an additional therapeutic agent, the compound is administered in close enough time to have the desired therapeutic effect. For example, the time period between each administration that can produce the desired therapeutic effect can be from several minutes to several hours, and can be determined based on the properties of each compound (e.g., efficacy, solubility, bioavailability, plasma half-life, and kinetic profile). For example, the compound of the invention and the second therapeutic agent can be administered in any order within 24 hours of each other, within 16 hours of each other, within 8 hours of each other, within 4 hours of each other, within 1 hour of each other, or within 30 minutes of each other.

[0358] More specifically, a first therapy (e.g., a prophylactic or therapeutic agent, such as a compound of the invention) can be administered to a subject prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concurrently with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second therapy (e.g., a prophylactic or therapeutic agent, such as an anti-cancer agent).

[0359] It is understood that the method of co-administering a first amount of a compound of the present invention and a second amount of an additional therapeutic agent can produce an enhanced or synergistic therapeutic effect, wherein the combined effect is greater than the additive effect produced by separate administration of the first amount of the compound of the present invention and the second amount of the additional therapeutic agent.

[0360] The term "synergistic" as used herein refers to a combination of a compound of the invention and another treatment (e.g., a prophylactic or therapeutic agent) that is more effective than the additive effect of the treatment. The synergistic effect of a combination of treatments (e.g., a combination of prophylactics or a combination of therapeutics) can allow the use of lower doses of one or more treatments and / or less frequent administration of the treatment to a subject. The ability to use lower doses of a treatment (e.g., a prophylactic or therapeutic agent) and / or less frequent administration of the treatment can reduce the toxicity associated with administering the treatment to a subject without reducing the effectiveness of the treatment in preventing, controlling or treating a condition. In addition, synergy can result in an improved effectiveness of a medicament in preventing, managing or treating a condition. Finally, the synergistic effect of a combination therapy (e.g., a combination of prophylactics or a combination of therapeutics) can avoid or reduce adverse or undesirable side effects associated with the use of any one treatment alone.

[0361] The presence of synergy can be determined using appropriate methods for evaluating drug interactions. Suitable methods include, for example, the Sigmoid-Emax equation (Holford, NHG and Scheiner, LB, Clin. Pharmacokinet. 6: 429-453 (1981)), the Loewe additivity equation (Loewe, S. and Muischnek, H., Arch. Exp. Pathol Pharmacol. 114: 313-326 (1926)) and the median effect equation (Chou, TC and Talalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). Each of the above-mentioned equations can be applied together with experimental data to generate corresponding graphs to help evaluate the effects of drug combinations. The corresponding graphs associated with the above-mentioned equations are concentration-effect curves, isobologram curves, and combination index curves, respectively.

[0362] In one aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer, a pharmaceutically acceptable excipient, and at least one additional therapeutic agent. In some embodiments, the compound of the present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer, and at least one additional therapeutic agent are co-formulated. In some embodiments, the compound of the present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer, and at least one additional therapeutic agent are separately formulated.

[0363] In another aspect, the present invention provides a kit comprising a compound, a pharmaceutically acceptable salt, a solvate, a hydrate, a tautomer or an optical isomer of the present invention and at least one additional therapeutic agent. The kit may include instructions for administering a compound, a pharmaceutically acceptable salt, a solvate, a hydrate, a tautomer or an optical isomer of the present invention and at least one additional therapeutic agent to a subject in need thereof.

[0364] In another aspect, the present invention provides a combination therapy for use as a medicament, wherein the combination therapy comprises administering to a subject in need thereof a compound, pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer of the present invention, and administering to a subject in need thereof an additional therapeutic agent.

[0365] The compound, pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer of the present invention and the additional therapeutic agent may be administered to a subject separately, sequentially or simultaneously.

[0366] Pharmaceutical compositions, kits and / or combination therapies can be used to treat gastrointestinal disorders or pulmonary diseases or disorders. Gastrointestinal disorders can be selected from constipation disorders, constipation-dominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergies or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastric and intestinal damage, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrheal diseases, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease and ischemic colitis. Pulmonary diseases or disorders can be selected from chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperresponsive airway disorders and idiopathic pulmonary fibrosis.

[0367] At least one additional therapeutic agent may be selected from aminosalicylate, corticosteroid, immunomodulator and combination thereof. Aminosalicylate is also known as 5-aminosalicylate (5-ASA). At least one additional therapeutic agent may be aminosalicylate. Aminosalicylate may be mesalamine. Aminosalicylate may be sulfasalazine. At least one additional therapeutic agent may be a corticosteroid. A corticosteroid may be budesonide. At least one additional therapeutic agent may be an immunomodulator. An immunomodulator may be a thiopurine. An immunomodulator may be methotrexate.

[0368] Pharmaceutical composition

[0369] While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (eg, formulation).

[0370] Therefore, in another embodiment of the present invention, there is provided a pharmaceutical composition comprising at least one compound of formula (1) as defined above and at least one pharmaceutically acceptable excipient.

[0371] When the pharmaceutical composition comprises at least one additional therapeutic agent, the compound of the present invention and the at least one additional therapeutic agent may be co-formulated or the compound of the present invention and the at least one additional therapeutic agent may be formulated separately.

[0372] The composition may be a tablet composition.

[0373] The composition may be a capsule composition.

[0374] Pharmaceutically acceptable excipients may be selected, for example, from carriers (e.g., solid, liquid or semisolid carriers), excipients, diluents (e.g., solid diluents, such as fillers or extenders; and liquid diluents, such as solvents and co-solvents), granulating agents, binders, glidants, coating agents, release controlling agents (e.g., release-delaying or -retarding polymers or waxes), adhesives, disintegrants, buffers, lubricants, preservatives, antifungal and antibacterial agents, antioxidants, buffers, tonicity modifiers, thickeners, flavoring agents, sweeteners, pigments, plasticizers, taste-masking agents, stabilizers or any other excipients conventionally used in pharmaceutical compositions.

[0375] The term "pharmaceutically acceptable" as used herein means compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for contact with the tissues of subjects (e.g., human subjects) without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio. Each excipient must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

[0376] Pharmaceutical compositions containing compounds of formula (1) can be formulated according to known techniques, see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.

[0377] The pharmaceutical composition may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ocular, otic, rectal, intravaginal or transdermal administration.

[0378] Suitable pharmaceutical dosage forms for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), caplets, pills, lozenges, syrups, solutions, powders, granules, elixirs and suspensions, sublingual tablets, wafers or patches, such as a buccal patch.

[0379] Tablet compositions may contain a unit dose of active compound and an inert diluent or carrier (e.g., a sugar or sugar alcohol (e.g., lactose, sucrose, sorbitol, or mannitol); and / or a non-sugar derived diluent (e.g., sodium carbonate, calcium phosphate, calcium carbonate), or a cellulose or derivative thereof (e.g., microcrystalline cellulose (MCC), methylcellulose, ethylcellulose, hydroxypropylmethylcellulose), and starch (e.g., corn starch)). Tablets may also contain standard ingredients such as binders and granulating agents, such as polyvinylpyrrolidone, disintegrants (e.g., swellable cross-linked polymers such as cross-linked carboxymethylcellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphate or citrate buffers), and effervescent agents (e.g., citrate / bicarbonate mixtures). Such excipients are well known and need not be discussed in detail herein.

[0380] Tablets may be designed to release the drug upon contact with gastric fluid (immediate release tablets), or in a controlled manner over a longer period of time (controlled release tablets), or upon contact with a specific area of ​​the GI tract.

[0381] The pharmaceutical composition generally comprises from about 1% (w / w) to about 95%, preferably % (w / w) of active ingredient and 99% (w / w) to 5% (w / w) of a pharmaceutically acceptable excipient (e.g. as defined above) or a combination of such excipients. Preferably, the composition comprises from about 20% (w / w) to about 90% (w / w) of active ingredient and 80% (w / w) to 10% of a pharmaceutical excipient or a combination of excipients. The pharmaceutical composition comprises from about 1% to about 95%, preferably from about 20% to about 90% of active ingredient. The pharmaceutical composition according to the invention may be, for example, in unit dosage form, for example in the form of ampoules, vials, suppositories, prefilled syringes, dragees, powders, tablets or capsules.

[0382] Tablets and capsules may contain, for example, 0% to 20% disintegrants, 0% to 5% lubricants, 0% to 5% glidants and / or 0% to 99% (w / w) fillers and / or bulking agents (depending on the drug dose). They may also contain 0% to 10% (w / w) polymer binders, 0% to 5% (w / w) antioxidants, 0% to 5% (w / w) pigments. In addition, sustained-release tablets typically contain 0% to 99% (w / w) controlled-release (e.g., delayed-release) polymers (depending on the dose). The film coating of a tablet or capsule typically contains 0% to 10% (w / w) polymers, 0% to 3% (w / w) pigments and / or 0% to 2% (w / w) plasticizers.

[0383] Parenteral formulations typically contain 0% to 20% (w / w) buffer, 0% to 50% (w / w) cosolvent and / or 0% to 99% (w / w) water for injection (WFI) (depending on the dose and whether lyophilized). Formulations for intramuscular depots may also contain 0% to 99% (w / w) oil.

[0384] Pharmaceutical preparations may be provided to patients in "patient packs" which contain an entire course of treatment in a single package, usually a blister pack.

[0385] The compound of formula (1) is generally present in a unit dosage form and therefore generally contains enough compound to provide the desired level of biological activity. For example, the formulation may contain 1 nanogram to 2 grams of active ingredient, such as 1 nanogram to 2 milligrams of active ingredient. Within these ranges, specific subranges of the compound are 0.1 milligram to 2 grams of active ingredient (more generally 10 milligrams to 1 gram, such as 50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (e.g., 1 microgram to 10 milligrams, such as 0.1 milligram to 2 milligrams of active ingredient).

[0386] For oral compositions, unit dosage forms may contain 1 mg to 2 g, more usually 10 mg to 1 g, for example 50 mg to 1 g, for example 100 mg to 1 g, of active compound.

[0387] The active compound is administered to a patient (eg, a human or animal patient) in need thereof in an amount sufficient to achieve the desired therapeutic effect (effective amount). The precise amount of compound administered can be determined by the attending physician according to standard procedures.

[0388] Process for preparing the compound of formula (I)

[0389] General synthetic scheme

[0390] Solution 1

[0391]

[0392] Referring to Scheme 1, the compound of formula G-5 can be synthesized from a proline derivative of formula G-1a (wherein PG is a protecting group, such as BOC). The amine compound of formula G-2 is coupled with G-1a, and then the amine protecting group PG is deprotected to provide an amide of formula G-3. Common coupling agents for achieving this transformation are well known to those skilled in the art, such as EDC, DCC, BOP, etc. The compound of formula G-3 can be reacted with a benzyl compound of formula G-4a via SN-2 reaction conditions to produce a compound of formula G-5 (wherein bromide is a leaving group), or can be reacted with an aldehyde compound of formula G-4b via reductive amination conditions to produce a compound of formula G-5.

[0393] Solution 2

[0394]

[0395] Referring to Scheme 2, compounds of the general formula G-5 can also be prepared from proline derivatives of the general formula G-1b (wherein the carboxylic acid of G-1b is protected by a simple ester such as a methyl ester). Compounds of the general formula G-8 can be synthesized by reacting G-1b with a benzyl compound of the general formula G-4a via SN-2 conditions (wherein bromide is a leaving group), or by reacting G-1b with an aldehyde compound of the general formula G-4b via reductive amination conditions. Compounds of the general formula G-8 can then be coupled with amine compounds of the general formula G-2 using the above-mentioned coupling conditions to provide compounds of the general formula G-5.

[0396] Solution 3

[0397]

[0398] Referring to Scheme 3, the compound of formula (1) can be synthesized from an organic boronic ester compound of formula G-6 by palladium-catalyzed coupling conditions using a palladium catalyst known to those skilled in the art (e.g., PdCl2(dppf)DCM). The compound of formula G-6 can be coupled with an aryl bromide compound of formula G-4a or a benzaldehyde compound of formula G-4b to produce a compound of formula G-7a or formula G-7b, respectively. The benzyl compound of formula G-7a (wherein the bromide is a leaving group) reacts with an amine compound of formula G-3 via SN-2 reaction conditions to produce a compound of formula (1). The benzaldehyde compound of formula G-7b reacts with an amine compound of formula G-3 via reductive amination conditions to produce a compound of formula (1).

[0399] Solution 4

[0400]

[0401] Referring to Scheme 4, using the above palladium-catalyzed coupling chemistry, a compound of formula G-5 can be coupled with an organoboronate compound of formula G-6 to provide a compound of formula (1). It is also well known to those skilled in the art that some compounds of formula (1) can be further derivatized. For example, a compound of formula (1) (wherein R 7 The OH group can be further derivatized by reaction with a suitable electrophile, such as sulfonyl chloride to produce sulfamoyloxy derivatives, carbamoyl chloride to produce carbamate derivatives, and the like.

[0402] Chromatographic methods

[0403] LC / MS Method A

[0404] Instrument: Acquity UPLC with photodiode array detector and QDA mass detector; Column: Acquity C-18, 1.6 micron, 50×1.6 mm; Gradient [time (min) / solvent B in A (%)]: 0.00 / 3, 0.20 / 3, 2.70 / 98, 3.00 / 100, 3.50 / 100, 3.51 / 3, 4.00 / 3; Solvents: Solvent A = 0.1% formic acid in water; Solvent B = 0.1% formic acid in water / acetonitrile (10:90); Column temperature 35° C.; Flow rate 0.9 mL / min.

[0405] LC / MS Method B&C

[0406] Instrument: HP 1100 with G1315A DAD, Waters Micromass ZQ; Column: Phenomenex Gemini-NX C-18, 3 micron, 2.0×30 mm; Method B gradient [time (min) / solvent B in A (%)]: 0.00 / 2, 0.10 / 2, 2.50 / 95, 3.50 / 95; Method C gradient [time (min) / solvent B in A (%)]: 0.00 / 2, 0.01 / 2, 8.40 / 95, 10.00 / 95; Solvent: Solvent A = 2.5 L H2O + 2.5 mL 28% ammonia in H2O solution; Solvent B = 2.5 L MeCN + 135 mL H2O + 2.5 mL 28% ammonia in H2O solution. Injection volume 1 μL; UV detection 230 to 400 nm; mass detection 130 to 800 AMU; column temperature 45°C; flow rate 1.5 mL / min.

[0407] LC / MS Method D

[0408] Instrument: Agilent 1260 Infinity LC with diode array detector, Agilent 6120B single quadrupole MS with API-ES source; Column: Restek pentafluorophenyl propyl, 3 micron, 2.1×30 mm. Gradient [time (min) / solvent B in A (%)]: 0.00 / 2, 0.1 / 2, 8.4 / 95, 10 / 95, 10.1 / 2, 12 / 2; Solvent: Solvent A = water (2.5 L) containing 2.5 mL formic acid; Solvent B = MeCN (2.5 L) containing 125 mL water and 2.5 mL formic acid. Injection volume 0.5 μL; UV detection 190 to 400 nm; Mass detection 130 to 800 AMU; Column temperature 40° C.; Flow rate 1.5 mL / min.

[0409] LC / MS Method E

[0410] Instrument: Agilent Technologies 1290 series with binary pump, diode array detector and G6120A quadrupole MS. Column: Agilent Poroshell 120EC-C18, 2.7 μm, 4.6×50 mm; Gradient [time (min) / solvent B in A (%)]: 0.00 / 10, 0.50 / 10, 4.00 / 90, 4.50 / 100, 4.51 / 10, 5.00 / 10; Solvent: Solvent A = 0.05% formic acid in water; Solvent B = 0.05% formic acid in acetonitrile; Column temperature 35° C.; Flow rate 1.0 mL / min.

[0411] LC / MS Method F

[0412] Instrument: Water 2690 with photodiode array detector and QDA mass detector, column: X-Bridge C-18, 5 micron, 100×1.6 mm; gradient [time (min) / solvent B in A (%)]: 0.01 / 10, 1.00 / 10, 5.00 / 100, 7.00 / 100, 7.50 / 10, 8.00 / 10; solvent: solvent A = 0.1% formic acid and 10 mM ammonium carbonate in water; solvent B = acetonitrile; column temperature 35°C; flow rate 0.9 mL / min.

[0413] LC / MS Method G

[0414] Instrument: Water 2690 with 996 photodiode array detector and Micromass ZQ mass detector; Column: C-18, 3.5 micron, 50×4.6 mm; Gradient [time (min) / solvent B in A (%)]: 0.00 / 10, 1.00 / 10, 4.00 / 100, 6.00 / 100, 6.50 / 10, 7.00 / 10; Solvents: Solvent A = 10 mM ammonium bicarbonate in water; Solvent B = methanol; Flow rate 1.2 mL / min.

[0415] LC / MS Method H

[0416] Instrument: Agilent 1290RRLC with Agilent 6120 mass detector; Column: X-Bridge C18 50*4.6 mm, 3.5 μm; Gradient: 95:5 at 0.01 min, 15:85 at 2.8 min, 5:95 at 3.5 min until 5.0 min, 95:5 at 5.01 min up to 6.0 min; Solvent: Solvent A = 5 mM ammonium bicarbonate in water; Solvent B = acetonitrile; Flow rate 1 mL / min.

[0417] LC / MS Method I

[0418] Instrument: Agilent 1290RRLC with Agilent 6120 mass detector; Column: BEH C18 2.1X50mm, 1.7μm; Gradient: 98:2 at 0.01min up to 0.5min, 30:70 at 3.0min (flow rate: 0.45ml / min), 5:95 at 4.0min to 5.5min (flow rate: 0.50ml / min), 98:2 at 5.51min up to 6.0min (flow rate: 0.45ml / min); Solvents: Solvent A = 5mM ammonium acetate & 0.1% FA in water; Solvent B = 0.1% FA in acetonitrile; Flow rate 0.45mL / min.

[0419] LC / MS Method J

[0420] Instrument: Waters Acquity H-class LCMS, Mass Lynx software, PDA detector and QDa mass detector; column: Gemini-NX C18, 3μm, 30×2mm; gradient: [time (min) / %A:%B] 3 min run: [0.00 / 100:0], [1.30 / 0:100], [1.55 / 0:100], [1.60 / 100:0], [3.00 / 100:0]; solvent: (A): 50mM ammonium acetate in water at pH 7.40 (B): acetonitrile; column temperature 40°C; flow rate 0.5mL / min; mass spectrometry solvent: 0.1% formic acid in 90% v / v methanol:water; injection 1.0μL; λ range 200nm to 500nm.

[0421] LC / MS Method K

[0422] Instrument: Waters Acquity H-class and SQ mass detector; Column: BEH C18 2.1X 50mm, 1.7μm; Gradient: 95:5 (flow rate: 0.55ml / min) at 0.01min up to 0.6min, 30:70 (flow rate: 0.60ml / min) at 0.6min, 10:90 (flow rate: 0.65ml / min) at 0.80min, 0:100 (flow rate: 0.65ml / min) at 1.1min up to 1.70min, 95:5 (flow rate: 0.55ml / min) at 1.71min up to 2.0min; Solvents: Solvent A = 2mM ammonium acetate and 0.1% FA in water; Solvent B = 0.1% FA in acetonitrile; Flow rate 0.55mL / min.

[0423] LC / MS Method L

[0424] Instrument: Waters Acquity H grade with SQ mass detector; Column: BEH C18 2.1X 50mm, 1.7μm; Gradient: 98:2 at 0.01 min up to 0.5 min, 30:70 at 3.0 min (flow rate: 0.45ml / min), 5:95 at 4.0 min to 5.5 min (flow rate: 0.50ml / min), 98:2 at 5.51 min up to 6.0 min (flow rate: 0.45ml / min); Solvents: Solvent A = 5mM ammonium acetate & 0.1% FA in water; Solvent B = 0.1% FA in acetonitrile; Flow rate 0.45mL / min.

[0425] Preparative HPLC purification

[0426] Where indicated, intermediates or final compounds were purified by reverse phase preparative HPLC conditions using the instrumentation and methods given below:

[0427] HPLC Method A

[0428] Gilson semi-preparative HPLC system - including 321 pump, 171 diode array detector and GX-271 liquid handler and Gilson Trilution software. Column Phenomenex Kinetix C18, 100×30 mm, 5 μm. Flow rate 30 mL / min, gradient of solvent B in solvent A: 5% to 35% solvent B (in 10 min), 100% solvent B (for 2 min); solvent A: water with 0.1% TFA. Solvent B: MeCN].

[0429] HPLC Method B

[0430] Gilson semi-preparative HPLC system - containing 321 pump, 171 diode array detector and GX-271 liquid handler and Gilson Trilution software. Column Gemini-NX C18, 100×30 mm, 5 μm, flow rate 30 mL / min. Gradient of solvent B in solvent A: 5% to 35% solvent B (in 10 min), 100% solvent B (for 2 min); Solvent A: water with 0.2% of 28% ammonia solution. Solvent B: MeCN].

[0431] HPLC Method C

[0432] Gilson semi-preparative HPLC system - contains dual piston pumps 331 and 332, 171 diode array detector and GX-271 liquid handler and Gilson Trilution software. Column Gemini-NX C18, 100×30 mm, 5 μm. Flow rate 30 mL / min, gradient of solvent B in solvent A: 5% to 35% solvent B (in 10 min), 100% solvent B (for 2 min); solvent A: water with 0.2% of 28% ammonia solution. Solvent B: MeCN].

[0433] HPLC Method D

[0434] Agilent Preparative 1200infinity Series

[0435] Column Sunfire C18 250×19 mm 5 μm. Flow rate 17 mL / min, gradient of solvent B in solvent A: 0.00 / 10, 17.00 / 16, 17.01 / 98, 19.00 / 98, 19.01 / 10, 21 / 10. Solvent A: water containing 0.1% FA, solvent B: MeCN.

[0436] HPLC Method E

[0437] SHIMADZU Preparative HPLC System - Contains LC-20AP pump, SPD-20A detector and Labsolutions (version 5.90) ​​software. Column: Agilent 10, Preparative-C18, 250 x 21.2 mm. Solvent / Gradient: 10% to 80% acetonitrile in water containing 0.1% TFA. Flow rate: 20 mL / min.

[0438] HPLC Method F

[0439] Shimadzu LC-20AP and UV detector. The column used was sunfire c18 (250*19) mm, 5μ. The column flow was 12.0 ml / min. The mobile phases used were (A) 0.1% formic acid in water and (B) 100% acetonitrile. The gradient solvent B was 0% to 20% in 30 minutes, 100% in 2 minutes, and then 100% to 0% in 5 minutes.

[0440] HPLC Method G

[0441] Waters Xbridge C 18 150*50mm*10um; Mobile phase: [water (10mM NH4HCO3) to ACN]; B%: 33% to 63%, min.

[0442] HPLC Method H

[0443] Phenomenex luna C 18 150*25mm*10um; Mobile phase: [water (0.225% FA) to ACN]; B%: 3% to 33%, 10 minutes.

[0444] HPLC Method I

[0445] Shimadzu LC-20AP and UV detector. The column used was sunfire c18 (250*19) mm, 5 microns, and the column flow rate was 14.0 ml / min. The mobile phases used were (A) 0.1% formic acid in water and (B) 100% acetonitrile. The gradient solvent B was 0% to 20% in 23 minutes, then 20% to 20% in 2 minutes, 100% in 2 minutes, and then 100% to 0% in 6 minutes.

[0446] Example

[0447] The compounds of formula (1) can be prepared according to synthetic methods known to those skilled in the art. The following examples are provided to provide a more complete understanding of the present invention and are not intended to limit the present invention in any way.

[0448] Where preparation routes are not included, the relevant intermediates are commercially available. Commercial reagents were used without further purification. Final compounds and intermediates were named using ChemDraw Professional, version 17.0.0.206 (121). Room temperature (RT) refers to about 20°C to 27°C. 1 H NMR spectra were recorded at 400 or 500 MHz on Bruker, Varian or Jeol instruments. Chemical shift values ​​are expressed in parts per million (ppm), i.e. (δ), relative to deuterated solvents such as chloroform-d (7.26 ppm), DMSO-d6 (2.50 ppm) or methanol-d4 (3.31 ppm). The following abbreviations are used for the multiplicity of NMR signals: s = singlet, br = broad, d = doublet, t = triplet, q = quartet, m = multiplet. Coupling constants are listed as J values, measured in Hz. NMR and mass spectrometry results are corrected to account for background peaks. Chromatography refers to the use of a 60 to 120 mesh or 40 to 633 μm, Column chromatography was performed on silica gel and under nitrogen pressure (flash chromatography) or automated flash chromatography using a Biotage Isolera apparatus. Microwave mediated reactions were performed in a Biotage initiator or a CEM Discover microwave reactor.

[0449] Synthesis of intermediates

[0450] Intermediate 1: 4-((S)-1-((R)-pyrrolidine-2-amido)ethyl)benzoic acid methyl ester hydrochloride (Intermediate 1-HCl)

[0451]

[0452] Step (i): To a mixture of N-Boc-D-proline (2.0 g, 9.29 mmol) in DMF (43.85 mL) was added diisopropylethylamine (4.82 mL, 27.87 mmol), HATU (4.25 g, 11.15 mmol) and methyl 4-[(1S)-1-aminoethyl]benzoate (2.0 g, 11.15 mmol). The mixture was stirred at room temperature overnight, after which the mixture was diluted with EtOAc and water (1:1) and the organic layer was separated. The organic phase was washed with brine (2×50 mL), dried (frit), and then concentrated in vacuo. The residue was purified by flash column chromatography (normal phase, 25 g SNAP KP-Sil, 70 mL / min, gradient: 0% to 100% ethyl acetate in isohexane) to give tert-butyl (2R)-2-[[(1S)-1-(4-methoxycarbonylphenyl)ethyl]carbamoyl]pyrrolidine-1-carboxylate (3.383 g, 9.0 mmol, 97% yield) as a light yellow solid. LC / MS (Method B) 2.05 min [M+H] + 377.

[0453] Step (ii): To a solution of (2R)-2-[[(1S)-1-(4-methoxycarbonylphenyl)ethyl]carbamoyl]pyrrolidine-1-carboxylic acid tert-butyl ester (4.03 g, 10.71 mmol) in 1,4-dioxane (20.2 mL) was added 4M HCl in 1,4-dioxane (20.2 mL, 10.71 mmol) which was stirred at room temperature for 3 hours. Once complete (monitored by LC / MS), 1,4-dioxane (20.2 mL) was added to the reaction mixture. The reaction mixture was then concentrated in vacuo to give the intermediate 1-HCl (3.39 g, 101%) as a white solid. LC / MS (Method B) 1.73 min [M+H] + 277.

[0454] Intermediate 2: Methyl 4-((S)-1-((R)-1-(3-bromobenzyl)pyrrolidine-2-amido)ethyl)benzoate (Intermediate 2)

[0455]

[0456] To a solution of intermediate 1-HCl (1.0 g, 3.2 mmol) in MeCN (96 mL) was added potassium carbonate (1.1 g, 7.99 mmol) and 3-bromobenzyl bromide (1.6 g, 6.39 mmol). The resulting mixture was heated to 70 ° C overnight, and then filtered through a phase separator, washed with EtOAc, and then concentrated in vacuo. The residue was dissolved in EtOAc, and the organic phase was washed with water and brine, dried over a hydrophobic glass frit, and concentrated in vacuo. The residue was purified by flash column chromatography (10 g SNAP KP-Sil, 30 mL / min, gradient: 30% to 100% ethyl acetate in isohexane) to give Intermediate 2 as a white solid (1.18 g, 83%). LC / MS (Method B) 1.73 min [M+H] + 445.

[0457] Intermediate 3: 4-((S)-1-((2R,4R)-4-hydroxypyrrolidine-2-amide)ethyl)benzoic acid methyl ester hydrochloride (Intermediate 3)

[0458]

[0459] Intermediate 3-HCl was synthesized in a similar manner to Intermediate 1 using (2R,4R)-1-tert-butoxycarbonyl-4-hydroxy-pyrrolidine-2-carboxylic acid. LC / MS (Method B) 1.47 min [M+H] + 293.

[0460] Intermediate 4: 4-((S)-1-((2R,4R)-1-(3-bromobenzyl)-4-hydroxypyrrolidine-2-amide)ethyl)benzoic acid methyl ester (Intermediate 4)

[0461]

[0462] Intermediate 4 was synthesized in a similar manner to Intermediate 2 using Intermediate 3-HCl. LC / MS (Method B) 2.17 min [M+H] + 463.

[0463] Intermediate 5: Methyl 4-((S)-1-((R)-1-(3-bromo-4-methoxybenzyl)pyrrolidine-2-amido)ethyl)benzoate (Intermediate 5)

[0464]

[0465] To the mixture of intermediate 1-HCl (300mg, 0.96mmol) and 3-bromo-4-methoxybenzaldehyde (412.49mg, 1.92mmol) in DCM (5mL) add sodium triacetoxyborohydride (426.87mg, 2.01mmol). The mixture is stirred at room temperature for 16 hours, then diluted with saturated NaHCO. The organic layer is separated, washed with salt water, dried (by passing through a hydrophobic glass frit), and concentrated. The residue is purified by flash column chromatography (normal phase, 10g SNAP KP-Sil, 30 mL / min, gradient: 30% to 100% ethyl acetate in isohexane) to give intermediate 5 (380 mg, 0.80 mmol, 83% yield) as a clear gum which was sonicated in Et2O to give a fluffy white solid. LC / MS (Method B) 2.36 min [M+H] + 475.

[0466] Intermediate 6: Methyl 4-((S)-1-((R)-1-(3-bromo-5-methoxybenzyl)pyrrolidine-2-amido)ethyl)benzoate (Intermediate 6)

[0467]

[0468] To a solution of intermediate 1-HCl (300 mg, 0.96 mmol) in MeCN (10.8 mL) was added 1-bromo-3-(bromomethyl)-5-methoxybenzene (0.15 mL, 1.92 mmol) and potassium carbonate (331.4 mg, 2.4 mmol). The resulting mixture was heated to reflux for 16 hours before being partitioned between EtOAc and water. The organic layer was separated, washed with brine, dried (frit) and concentrated. The residue was purified by flash column chromatography (25 g Biotage SNAP KP-Sil 60 mL / min, gradient: 0% to 90% EtOAc in isohexane) to give intermediate 6 (300 mg, 0.63 mmol, 66% yield) as a white solid. LC / MS (Method B) 2.44 min [M+H] + 475.

[0469] Intermediate 7: (3-bromobenzyl)-D-proline (Intermediate 7)

[0470]

[0471] Step (i): D-proline methyl ester hydrochloride (10.00 g, 60.38 mmol) was dissolved in ACN (120 mL) and Na2CO3 (12.80 g, 120.77 mmol) was added and the reaction mixture was stirred at room temperature for 15 minutes. 1-Bromo-3-(bromomethyl)benzene (18.11 g, 72.46 mmol) was then added and the reaction mixture was stirred at 800°C for 16 hours. The reaction mixture was then partitioned between water (300 mL) and EtOAc (200 mL) and the aqueous layer was extracted again with EtOAc (2 x 100 mL). The organic layers were combined and dried (Na2SO4), the solvent was removed in vacuo and the crude product was purified by gradient column chromatography. The product was eluted at 0% to 20% EtOAc in hexanes to give pure methyl (3-bromobenzyl)-D-proline (11.00 g, 61.34%) as a yellow viscous substance. LC / MS: (Method A): m / z 298 [M+H] + In 1.35 minutes.

[0472] Step (ii): (3-bromobenzyl)-D-proline methyl ester (10.60 g, 35.68 mmol) was dissolved in dioxane (50 mL) and water (30 mL), LiOH monohydrate (7.48 g, 178.42 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 6 hours. Glacial acetic acid (20 mL) was then added to bring the solution to pH about 6, and the reaction mixture was then concentrated in vacuo to obtain the crude product, which was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica), eluting the product at 0% to 13% MeCN in water (0.1% FA as a modifier) ​​to give pure ((3-bromobenzyl)-D-proline) (7.8 g, 77.%) as a white solid. LC / MS: (Method A): m / z 284 [M+H] + In 1.28 minutes.

[0473] Intermediate 8: (R)-methyl 4-((1-(3-bromobenzyl)pyrrolidine-2-amido)methyl)-2-hydroxybenzoate (Intermediate 8)

[0474]

[0475] Step (i): Intermediate 7 (2.50 g, 8.83 mmol) was dissolved in MeCN (15 mL), and methyl 4-(aminomethyl)-2-methoxybenzoate (1.90 g, 9.71 mmol) was added to the mixture at room temperature. Propylphosphonic anhydride (50% solution in EtOAC) (11.42 mL, 17.66 mmol) was then added, and the reaction mixture was stirred at room temperature for 30 minutes. TEA (3.88 mL, 26.54 mmol) was then added at 0 ° C, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was distributed between saturated NaHCO3 aqueous solution (100 mL) and EtOAc (100 mL). The water layer was further extracted with EtOAc (2 × 70 mL). The organic layers were combined and dried over Na2SO4, the solvent was removed in vacuo, and the crude product was purified by gradient flash column chromatography (normal phase, silica), eluting the product at 0% to 64% EtOAc in hexanes to give pure (R)-methyl 4-((1-(3-bromobenzyl)pyrrolidine-2-amido)methyl)-2-methoxybenzoate (2.50 g, 60%) as a yellow sticky substance. LC / MS: (Method A): m / z 461 [M+H] + , at 1.56 minutes.

[0476] Step (ii): (R)-methyl 4-((1-(3-bromobenzyl)pyrrolidine-2-amido)methyl)-2-methoxybenzoate (0.30 g, 0.65 mmol) was dissolved in DCM (30 mL) and the reaction mixture was cooled to -78 °C. BBr3 was added as a 1 M solution in DCM (3.3 mL, 3.26 mmol) and the reaction mixture was stirred at -78 °C for 1 hour. The reaction mixture was then partitioned between saturated aqueous NaHCO3 solution (70 mL) and DCM (70 mL) and the aqueous layer was extracted again with DCM (2 x 40 mL). The organic layers were combined and dried (Na2SO4) and the solvent was removed in vacuo to give pure intermediate 8 (0.25 g, 86%) as an off-white solid. LC / MS: (Method A): m / z 447 [M+H] + At 1.67 minutes.

[0477] Intermediate 9: (S)-4-(1-aminoethyl)-2-methoxybenzoic acid methyl ester (Intermediate 9)

[0478]

[0479] (S)-4-(1-aminoethyl)-2-methoxybenzoic acid (2.00 g, 10.25 mmol) was dissolved in methanol (20 mL) at room temperature. HCl was added as a 4N solution in dioxane (10 mL, 40 mmol) and the reaction mixture was stirred at 70 °C for 8 hours. The reaction mixture was concentrated in vacuo and the resulting residue was purified by trituration with diethyl ether to give intermediate 9 (2.1 g, quantitative) as a white solid. LC / MS: (Method A) No mass ions at 0.93 min.

[0480] Intermediate 10: 4-((S)-1-((R)-1-(3-bromobenzyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoic acid methyl ester (Intermediate 10)

[0481]

[0482] Step (i): Intermediate 9 (1.5 g, 7.71 mmol) was dissolved in MeCN (15 mL), and intermediate 7 (2.23 g, 7.89 mmol) was added to the reaction mixture at room temperature. HATU (4.09 g, 10.75 mmol) was then added and stirred for 30 minutes. N, N-diisopropylethylamine (2.50 mL, 14.34 mmol) was then added at 0 ° C, and stirred for 2 hours at room temperature. The reaction mixture was then distributed between saturated NaHCO3 aqueous solution (250 mL) and EtOAc (250 mL). The aqueous layer was further extracted with EtOAc (2×150 mL), and the organic layers were combined and dried over Na2SO4, the solvent was removed in vacuo, and the crude product was purified by gradient flash column chromatography (reverse phase, followed by normal phase). First, using MeCN in water as the mobile phase, the reverse phase flash column chromatography product was eluted at 0% to 68%. Second, the normal phase chromatography product was eluted using a gradient of 0% to 78% EtOAc in hexanes to afford methyl 4-((S)-1-((R)-1-(3-bromobenzyl)pyrrolidine-2-amido)ethyl)-2-methoxybenzoate (1.50 g, 44.11%) as a white solid. LC / MS (Method G): The product was identified, m / z 475.1 (ES+, M+2) at 1.438 min.

[0483] Step (ii): Methyl 4-((S)-1-((R)-1-(3-bromobenzyl)pyrrolidine-2-amido)ethyl)-2-methoxybenzoate (1.50 g, 3.16 mmol) was dissolved in DCM (30 mL) and the mixture was cooled to -78 °C. BBr3 was added as a 1 M solution in DCM (3.79 mL, 3.79 mmol) and the reaction mixture was stirred at -78 °C for 1 hour. The reaction mixture was then partitioned between saturated aqueous NaHCO3 (200 mL) and DCM (250 mL). The aqueous layer was extracted again with DCM (2 x 150 mL) and the organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 65% MeCN in water to give pure intermediate 10 as a white solid (0.99 g, 68%). LC / MS: (Method G): m / z 461 [M+H] + At 5.43 minutes.

[0484] Intermediate 11: 3'-(Bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 11)

[0485]

[0486] Step (i): 3-bromo-5-hydroxybenzaldehyde (4.00 g, 20.00 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (5.74 g, 22.00 mmol) and K2CO3 (3.20 g, 60.01 mmol) were dissolved in 1,4-dioxane (15 mL) and water (15 mL). Nitrogen was then purged through the mixture at room temperature for 20 minutes, followed by the addition of PdCl2(dppf)DCM (1.63 g, 2.00 mmol). The reaction mixture was then stirred at 80 ° C for 2 hours. The reaction mixture was then partitioned between water (400 mL) and EtOAc (250 mL), and the aqueous layer was further extracted with EtOAc (2×100 mL). The organic layers were combined and dried (Na2SO4), the solvent was removed in vacuo, and the crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) and the product was eluted at 0% to 35% MeCN in water to give 3'-formyl-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (3.60 g, 71%) as a brown solid. LC / MS: (Method A): m / z 256 [M+H] + , at 1.09 minutes.

[0487] Step (ii): 3'-Formyl-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.50 g, 5.88 mmol) was dissolved in methanol (15 mL) at room temperature. The reaction mixture was cooled to 0°C, sodium borohydride (0.44 g, 11.76 mmol) was added at 0°C, and then stirred at room temperature for 2 hours. The reaction mixture was partitioned between saturated aqueous NaHCO3 solution (100 mL) and EtOAc (80 mL), and the aqueous layer was further extracted with EtOAc (3×50 mL). The organic layers were combined and dried (Na2SO4), and the solvent was removed in vacuo to give 3'-hydroxy-5'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.36 g, 90%) as an off-white solid. LC / MS: (Method A): m / z 258 [M+H] + , at 0.94 minutes.

[0488] Step (iii): 3'-Hydroxy-5'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.36 g, 5.29 mmol) was dissolved in 33% HBr in CH3COOH (12 mL) at room temperature, and the resulting reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was then partitioned between saturated aqueous NaHCO3 solution (200 mL) and EtOAc (150 mL), and the aqueous layer was further extracted with EtOAc (2×50 mL). The organic layers were combined and dried (Na2SO4), and the solvent was removed in vacuo to give 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.59 g, 94%) as a reddish brown solid. LC / MS: (Method A): m / z 320 [M+H] + , at 1.23 minutes.

[0489] Intermediate 12: 3'-(Bromomethyl)-2-methyl-[1,1'-biphenyl]-4-sulfonamide (Intermediate 12)

[0490]

[0491] Step (i): 4-Bromo-3-methyl-benzenesulfonamide (1.00 g, 3.99 mmol), bis(pinacolato)diboron (2.02 g, 7.98 mmol) and KOAc (1.17 g, 11.97 mmol) were dissolved in dioxane (10 mL) at room temperature. Nitrogen was purged at room temperature for 30 minutes. After this, PdCl2(dppf)DCM (0.325 g, 0.399 mmol) was added and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was then partitioned between water (100 mL) and EtOAc (100 mL) and the aqueous layer was further extracted with EtOAc (2×100 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by gradient flash column chromatography (normal phase, silica) eluting the product at 0% to 30% EtOAc in hexanes to give pure 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (1.0 g, 85%) as a yellow solid. LC / MS: (Method F): The m / z of the product was not supported in the main peak (ES+) at 0.46 minutes.

[0492] Step (ii): 3-bromobenzaldehyde (4.50 g, 24.46 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (8.72 g, 29.35 mmol) and K2CO3 (10.20 g, 73.38 mmol) were dissolved in 1,4-dioxane (18 mL) and water (12 mL). Nitrogen was purged at room temperature for 30 minutes. Then, PdCl2(dppf)DCM (1.99 g, 2.44 mmol) was added and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was distributed between water (500 mL) and EtOAc (150 mL). The aqueous layer was further extracted with EtOAc (2×150 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by gradient column chromatography (normal phase, silica) eluting the product at 0% to 40% EtOAc in hexanes to give pure 3'-formyl-2-methyl-[1,1'-biphenyl]-4-sulfonamide (4.5 g, 67%) as an off-white solid. LC / MS: (Method A): The expected mass at 1.74 minutes for the main peak was not supported.

[0493] Step (iii): 3'-formyl-2-methyl-[1,1'-biphenyl]-4-sulfonamide (4.00 g, 14.54 mmol) was dissolved in methanol (30 mL) at room temperature. The reaction mixture was cooled to 0 ° C. and sodium borohydride (1.65 g, 43.62 mmol) was added in batches. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with cold water (50 mL). A solid precipitated, which was filtered and dried in vacuo to give 3'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-sulfonamide (3.6 g, 79%) as an off-white solid. LC / MS: (Method A): 260. [M+H-H2O] + , at 1.51 minutes.

[0494] Step (iv): 3'-(Hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-sulfonamide (3.60 g, 12.99 mmol) was dissolved in HBr (64% in water) (35 mL) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with cold water (50 mL). A solid precipitated, which was filtered and dried in vacuo to give 3'-(bromomethyl)-2-methyl-[1,1'-biphenyl]-4-sulfonamide (3.7 g, 84%) as a yellow solid. LC / MS: (Method A): Mass was not supported at 1.51 minutes.

[0495] Intermediate 13: 3'-(Bromomethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 13)

[0496] Intermediate 13 was synthesized by a method similar to Intermediate 12 using 4-bromo-3-methylbenzamide to give Intermediate 13 LC / MS: (Method A): m / z 305 [M+H] + , at 2.02 minutes.

[0497] Intermediate 14

[0498]

[0499] Step (i): To a solution of compound a1 (2.4 g, 11.16 mmol, 1 eq.) and compound a2 (1.62 g, 13.39 mmol, 1.2 eq.) in THF (50 mL) was added Ti(OEt)4 (5.09 g, 22.32 mmol, 4.63 mL, 2 eq.). The mixture was stirred at 30°C for 5 hours. TLC (PE:EA=5:1) indicated that compound a1 was completely consumed and a new spot was formed. The reaction mixture was diluted with H2O (50 mL) and extracted with EA (100 mL*2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g Silica flash column, elution with 0% to 30% ethyl acetate / petroleum ether gradient @50 mL / min) to afford compound a3 (3.5 g, 11.00 mmol, 98.55% yield) as a yellow oil, which was identified by 1H NMR.

[0500] 1 H NMR (CDCl3, 400MHz) δ8.54 (s, 1H), 7.67 (d, J=8.0Hz, 1H), 7.40 (d, J=1.6Hz, 1H), 7.30 (dd, J=1.6, 8.0Hz, 1H), 3.98 (s, 3H), 1.28 (s, 9H).

[0501] Step (ii): Dimethylzinc (1M, 13.36mL, 1.7 equivalents) was dissolved in THF (50mL), and MeMgBr (3M, 3.93mL, 1.5 equivalents) was added dropwise at 10°C, and the mixture was then stirred at 10°C for 30 minutes. The reaction solution was then cooled to 0°C, and a solution of compound a3 (2.50g, 7.86mmol, 1 equivalent) in THF (12.5mL) was added dropwise, and the reaction mixture was then stirred at 10°C for 1 hour. TLC (PE:EA=1:1) indicated that compound a3 was completely consumed. The reaction mixture was diluted with H2O (30mL) and extracted with EA (50mL*2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g Silica flash column, elution with 30% to 60% ethyl acetate / petroleum ether gradient @50 mL / min) to afford compound a4 (1.2 g, 3.52 mmol, 44.83% yield, 98.1% purity) as a white solid, which was identified by 1H NMR.

[0502] SFC: Rt = 1.220 min, ee: 97.1%.

[0503] 1 H NMR (CDCl3, 400MHz) δ7.49 (d, J=8.4Hz, 1H), 6.90 (s, 1H), 6.82 (d, J=8.0Hz, 1H), 4.55 (dd, J=2.0, 6.4Hz, 1H), 3.90 (s, 3H), 3.32 (s, 1H), 1.52 (d, J=6.8Hz, 3H), 1.22 (s, 9H).

[0504] Step (iii): TEA (1.27 g, 12.56 mmol, 1.75 mL, 3 eq.) and Pd (dppf) Cl2.CH2Cl2 (342.03 mg, 418.82 umol, 0.1 eq.) were added to a solution of compound a4 (1.4 g, 4.19 mmol, 1 eq.) in EtOH (40 mL). The mixture was stirred at 80 ° C for 16 hours under CO (50 PSI). LC-MS showed that compound a4 was completely consumed and a main peak with a desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=3 / 1 to 1 / 1) to obtain compound a5 (1.35 g, 4.09 mmol, 97.65% yield, 99.2% purity) as a yellow oil, which was determined by HNMR.

[0505] LCMS: [M+1] + =328.4.

[0506] SFC: Rt = 0.901 min, ee = 97.9%.

[0507] 1 H NMR (CDCl3, 400MHz) δ7.77 (d, J=7.6Hz, 1H), 7.02-6.90 (m, 2H), 4.63-4.57 (m, 1H), 4.36 (q, J=7.2Hz , 2H), 3.91 (s, 3H), 3.33 (d, J=1.6Hz, 1H), 1.54 (d, J=6.8Hz, 3H), 1.38 (t, J=7.2Hz, 3H), 1.23 (s, 9H).

[0508] Step (iv): At 0 ° C, HCl / dioxane (4M, 20 mL) was added to a solution of compound a5 (1.35 g, 4.12 mmol, 1 equivalent) in DCM (15 mL). The mixture was stirred at 15 ° C for 12 hours. LC-MS showed that compound a5 was completely consumed. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was ground with PE:EA=5:1 (10 mL) at 15 ° C for 10 minutes to obtain compound a6 (920 mg, 3.54 mmol, 85.91% yield, HCl) as a yellow solid, which was determined by HNMR.

[0509] 1 H NMR (DMSO-d6, 400MHz) δ8.67 (s, 3H), 7.64 (d, J=8.0Hz, 1H), 7.44 (s, 1H), 7.13 (d, J=8.0Hz, 1H), 4. 50-4.36 (m, 1H), 4.24 (q, J=6.8Hz, 2H), 3.85 (s, 3H), 1.52 (d, J=6.4Hz, 3H), 1.27 (t, J=7.2Hz, 3H).

[0510] Step (v): To a solution of compound a7 (795.58 mg, 3.70 mmol, 1.5 eq.) in DMF (8 mL) was added HATU (1.87 g, 4.93 mmol, 2 eq.) and DIEA (1.59 g, 12.32 mmol, 2.15 mL, 5 eq.), and the mixture was stirred at 25 °C for 0.5 h. Compound a6 (640 mg, 2.46 mmol, 1 eq., HCl) was added and the resulting mixture was stirred at 25 °C for 1 h. LC-MS showed that compound a6 was completely consumed and a main peak with the expected mass was detected. The reaction mixture was diluted with H2O (30 mL) and extracted with EA (50 mL*2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g Silica flash column, elution with 0% to 50% ethyl acetate / petroleum ether gradient @ 40 mL / min) to give compound a8 (1.26 g, crude) as a yellow oil, which was identified by HNMR.

[0511] LCMS: [M-Boc] + =321.5.

[0512] SFC: Rt = 0.926 min, ee: 100%.

[0513] 1H NMR (DMSO-d6, 400MHz) 88.39-8.23 (m, 1H), 7.56 (d, J=7.6Hz, 1H), 7.12 (s, 1H), 6.97 (d, J=7.6Hz, 1H), 5.04-4.81 (m, 1H), 4.23 (q, J=7.2Hz, 2H), 4.18 -4.05(m, 1H), 3.88-3.78(m, 3H), 3.49-3.35(m, 1H), 3.29-3.22(m, 1H), 2. 22-2.03(m, 1H), 1.95-1.71(m, 3H), 1.47-1.29(m, 8H), 1.24-1.11(m, 7H).

[0514] Step (vi): HCl / dioxane (4M, 10 mL) was added to a solution of compound a8 (1.2 g, 2.85 mmol, 1 equivalent) in DCM (10 mL). The mixture was stirred at 15 ° C for 1 hour. LC-MS showed that compound a8 was completely consumed and a main peak with the expected mass was detected. The reaction mixture was concentrated under reduced pressure to obtain intermediate 14 (830 mg, thick, HCl) as a yellow oil.

[0515] LCMS: [M+1] + =321.2.

[0516] Intermediate 15

[0517]

[0518] Step (i): A mixture of compound bl (1 g, 4.65 mmol, 1 eq.), NH4Cl (373.12 mg, 6.98 mmol, 1.5 eq.), HATU (2.65 g, 6.98 mmol, 1.5 eq.) and DIEA (1.20 g, 9.30 mmol, 1.62 mL, 2 eq.) in DMF (25 mL) was degassed and purged 3 times with N2, and then stirred at 30°C for 12 hours under N2 atmosphere. LCMS indicated that compound bl was completely consumed. The mixture was slowly added dropwise to water (35 mL). The precipitate was filtered and the filter cake was concentrated to give compound b2 (600 mg, 2.80 mmol, 60.28% yield) as a brown solid, which was determined by HNMR.

[0519] LCMS: m / z=213.7 [M+1].

[0520] 1H NMR (DMSO-d6, 400MHz) δ 8.00 (s, 1H), 7.85 (d, J = 1.6Hz, 1H), 7.67-7.65 (m, 1H), 7.62-7.59 (m, 1H), 7.42 (s, 1H), 2.38 (s, 3H).

[0521] Step (ii): A mixture of compound b2 (450 mg, 2.10 mmol, 1 eq.), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (640.60 mg, 2.52 mmol, 1.2 eq.), AcOK (412.63 mg, 4.20 mmol, 2 eq.), Pd(dppf)Cl2 (153.82 mg, 210.22 umol, 0.1 eq.) in dioxane was degassed and purged with N2 3 times, and then stirred at 85° C. for 12 hours under N2 atmosphere. LCMS indicated that compound b2 was completely consumed, and a main peak with the expected mass was detected. TLC (petroleum ether:ethyl acetate=1:1) indicated that compound b2 was completely consumed. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography ( 20g Purification was performed using silica flash column, elution with a gradient of 0% to 65% ethyl acetate / petroleum ether @ 45 mL / min) to afford compound b3 (440 mg, 1.69 mmol, 80.15% yield) as a white solid, which was identified by HNMR.

[0522] LCMS: m / z=261.9 [M+1].

[0523] 1 H NMR (CDCl3, 400MHz) δ7.82 (d, J=7.6Hz, 1H), 7.65-7.54 (m, 2H), 6.22-5.84 (m, 2H), 2.58 (s, 3H), 1.36 (s, 12H).

[0524] Step (iii): To a solution of compound b3 (1 g, 3.83 mmol, 1 eq.), compound b4 (1.07 g, 4.98 mmol, 1.3 eq.) in dioxane (30 mL) and H2O (3 mL) was added K2CO3 (1.06 g, 7.66 mmol, 2 eq.) and Pd(dppf)Cl2 (280.21 mg, 382.96 umol, 0.1 eq.). Under N2, the mixture was stirred at 80°C for 12 hours. TLC (PE:EA=1:1) indicated that compound b3 was completely consumed and a new spot was formed. The reaction mixture was diluted with H2O (30 mL) and extracted with EA (50 mL*2). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate=3 / 1 to 1 / 2) to give intermediate 15 (550 mg, 2.04 mmol, 53.33% yield) as a yellow solid which was identified by HNMR.

[0525] 1 H NMR (CDCl3, 400MHz) δ10.03 (s, 1H), 7.80 (s, 1H), 7.69 (d, J=4.4Hz, 1H), 7.42 (s, 2H) ), 7.33 (d, J=4.8Hz, 1H), 7.13 (s, 1H), 6.55-5.32 (m, 2H), 3.92 (s, 3H), 2.34 (s, 3H).

[0526] Intermediate 16: (S)-1-(4-bromo-3-methoxyphenyl)ethane-1-amine (Intermediate 16)

[0527]

[0528] Step (i): A solution of 4-bromo-3-methoxybenzaldehyde (31.16 g, 0.14 mol, 1 eq.) was dissolved in THF (240 mL) at room temperature, and then Ti(OEt)4 (66.10 g, 0.28 mol, 61.20 mL, 2 eq.) was added to the reaction mixture at room temperature, and the reaction mixture was stirred at room temperature for 15 minutes. (R)-2-methylpropane-2-sulfinamide (28.09 g, 0.23 mol, 1.6 eq.) pre-dissolved in THF (60 mL) was added to the reaction mixture, and the mixture was stirred at 30°C overnight. TLC (EA: hexane = 2: 8) indicated that 4-bromo-3-methoxybenzaldehyde was completely consumed and a new spot was formed. The reaction mixture was diluted with H2O (300 mL), but then diluted with salt, and filtered through a vacuum pump, and the filtrate was extracted with EA (3×500 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (normal phase silica, 0% to 15.0% EA / hexanes) to give (R,E)-N-(4-bromo-3-methoxybenzylidene)-2-methylpropane-2-sulfenamide (35.9 g, 77.85% yield) as a white solid.

[0529] TLC: (2.0:8.0, EtOAc / hexane, RF: 0.40).

[0530] Mass (ESI+ve): 318.0[M].

[0531] LCMS: 100.0% (LCMS method H), RT: 3.545 min, 280.0 nm.

[0532] 1 H NMR (400MHz, CDC13) δ: 8.57 (s, 1H), 7.69-7.71 (d, 1H), 7.44 (d, 1H), 7.30-7.33 (dd, 1H), 4.01 (s, 3H), 1.31 (s, 9H).

[0533] Step (ii): Dimethylzinc (1.5 M in toluene, 269.66 mL, 3.7 eq.) was dissolved in THF (150 mL), and MeMgBr (3.0 M in diethyl ether, 116.6 mL, 3.2 eq.) was added dropwise at 15 to 20° C., and the mixture was then stirred at 15 to 20° C. for 45 minutes. The reaction solution was then cooled to 0° C., and a solution of (R,E)-N-(4-bromo-3-methoxybenzylidene)-2-methylpropane-2-sulfenamide (34.79 g, 0.034 mol, 1 eq.) pre-dissolved in THF (150 mL) was added dropwise, and the reaction mixture was then stirred at 0° C. for 30 minutes. TLC (EA: hexane = 5:5) indicated that (R,E)-N-(4-bromo-3-methoxybenzylidene)-2-methylpropane-2-sulfenamide was completely consumed. The reaction mixture is diluted with cold H2O (1000mL) and then solid, then acidified with 1N HCl (Ph=2 to 3) and extracted with EA (800mL×3). The combined organic layer is dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue is purified by flash silica gel chromatography (normal phase silica, under 0% to 45.0% EA / hexane), to obtain (R)-N-((S)-1-(4-bromo-3-methoxyphenyl)ethyl)-2-methylpropane-2-sulfinamide (25.57g, 69.97% yield, 97.20% purity) as an off-white solid.

[0534] TLC: (5.0:5.0, EtOAc / hexane, RF: 0.25).

[0535] Mass (ESI+ve): 335.8[M+1].

[0536] LCMS: 100.0% (LCMS method H), RT: 3.067 min, 254.0 nm.

[0537] 1 H NMR (400MHz, CDCl3) δ: 7.50-7.52 (s, 1H), 7.01 (s, 1H), 6.82-6.84 (dd, 1H), 4 .77-4.78(s, 1H), 4.42(s, 1H), 3.93(s, 3H), 1.58-1.62(t, 6H), 1.29(s, 9H).

[0538] Step (iii): To a solution of (R)-N-((S)-1-(4-bromo-3-methoxyphenyl)ethyl)-2-methylpropane-2-sulfinamide (25.57 g, 0.07 mol, 1 eq.) in dioxane (20 mL) was added HCl in dioxane (4.0 M, 125 mL) at 0°C. The mixture was stirred at room temperature for 3 hours. TLC (EA:hexane=5:5) indicated that (R)-N-((S)-1-(4-bromo-3-methoxyphenyl)ethyl)-2-methylpropane-2-sulfinamide was completely consumed. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was washed with dioxane (40 mL) and concentrated under reduced pressure to give a residue to give (S)-1-(4-bromo-3-methoxyphenyl)ethane-1-amine (Intermediate 16) (18.73 g, HCl salt) as a white solid.

[0539] TLC: (5.0:5.0, EtOAc / hexane, RF: 0.25).

[0540] LCMS: 100.0% (LCMS method H), RT: 2.371 min, 230.0 nm.

[0541] 1 H NMR (400MHz, DMSO-D20) δ: 8.75 (s, 3H), 7.59-7.68 (d, 1H), 7.49 (s, 1H), 7.02 -7.04(d, 1H), 4.36-4.39(s, 1H), 3.88(s, 3H), 3.56(s, 1H), 1.51-152(t, 3H).

[0542] Intermediate 17: 4'-Hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-carbaldehyde (Intermediate 17)

[0543]

[0544] To a solution of (4-hydroxy-2-methylphenyl)boronic acid (3.5 g, 0.02 mol, 1 eq.), 3-bromo-5-methoxybenzaldehyde (4.95 g, 0.020 mol, 1.0 eq.) in dioxane (35 mL) and H2O (11 mL) was added K2CO3 (6.35 g, 0.046 mol, 2 eq.), and the mixture was degassed for 10 minutes. After degassing, Pd(dppf)Cl2 (1.6 g, 0.002 mol, 0.1 eq.) was added to the reaction mixture. The reaction mixture was then stirred at 80°C overnight. TLC (EA:hexane=5:5) indicated that (4-hydroxy-2-methylphenyl)boronic acid was completely consumed and a new spot was formed. The reaction mixture was diluted with H2O (300 mL) and extracted with EA (3×150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (normal phase silica, at 0% to 46.0% EA / hexanes) to give 4'-hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-carbaldehyde (Intermediate 17) (4.0 g, 71.68% yield) as a brown solid.

[0545] TLC: (5.0:5.0, EtOAc / hexane, RF: 0.25).

[0546] Mass (ESI+ve): 243.0[M+1].

[0547] LCMS: 95.66% (LCMS method H), RT: 3.029 min, 254.0 nm.

[0548] 1 H NMR (400MHz, DMSO-D20) δ: 10.01 (s, 1H), 9.48 (s, 1H), 7.43-7.36 (d, 2H), 7. 16(s, 1H), 7.08-7.06(d, 1H), 6.67-6.71(m, 2H), 3.86(s, 3H), 2.18(s, 3H).

[0549] Synthesis of compounds of formula I

[0550] Example 1: 4-((1S)-1-((2R)-1-((4'-hydroxy-2'-methyl-1',2',3',4',5',6'-hexahydro-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 1)

[0551]

[0552] Step (i): A mixture of intermediate 2 (150.Mg, 0.3400mmol), (4-hydroxy-2-methylphenyl)boric acid (56.3mg, 0.3700mmol), potassium carbonate (0.09g, 0.67mmol) and tetrakis(triphenylphosphine)palladium(0) (11.68mg, 0.01mmol) in 1,4-dioxane (1.4mL) and water (0.3mL) was heated to 100°C in a microwave reactor for 1 hour. (4-hydroxy-2-methylphenyl)boric acid (56.3mg, 0.3700mmol) was further added and the mixture was placed in a microwave reactor for another 1 hour. The crude product was diluted with water and EtOAc, and the organic layer was separated, washed with water, brine, and dried by passing through a hydrophobic glass frit, then concentrated in vacuo. The residue was purified by flash column chromatography (normal phase, 10g, SNAP KP-Sil, 30 mL / min, gradient: 30% to 100% ethyl acetate in isohexane) to give methyl 4-[(1S)-1-[[(2R)-1-[[3-(4-hydroxy-2-methyl-phenyl)phenyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (148 mg, 93%) as a light yellow gum. LC / MS (Method B) 2.36 min [M+H] + 473.

[0553] Step (ii): To a solution of methyl 4-[(1S)-1-[[(2R)-1-[[3-(4-hydroxy-2-methyl-phenyl)phenyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (148.mg, 0.31mmol) in 1,4-dioxane (2.62mL) and water (2.62mL) was added lithium hydroxide monohydrate (59.13mg, 1.41mmol). The reaction was stirred at room temperature for 3 hours, then checked for completion by LC / MS and then concentrated in vacuo. The crude residue was purified by reverse phase HPLC (HPLC Method A) to give Compound 1 (52.5mg, 36%) as a white solid. LC / MS (Method C): m / z 459 [M+H] + 2.30 minutes.

[0554] 1H NMR (400MHz, DMSO-d6) δ7.91 (m, 1H), 7.74 (m, 2H), 7.33 (dd, J=7.5Hz, 1H), 7.27-7.21 (m , 2H), 7.18-7.08 (m, 3H), 6.98 (m, 1H), 6.68-6.61 (m, 2H), 4.85-4.76 (m, 1H), 3.80 (d, J=1 2.9Hz, 1H), 3.52 (d, J=12.9Hz, 1H), 3.08 (dd, J=9.5, 4.1Hz, 1H), 2.99-2.93 (m, 1H), 2.39 -2.29 (m, 1H), 2.13 (s, 3H), 2.09-1.97 (m, 2H), 1.72-1.61 (m, 3H), 1.24 (d, J=7.0Hz, 3H).

[0555] No exchangeable acid protons were observed in the spectrum.

[0556] Example 2: 4-((S)-1-((R)-1-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 2)

[0557] Compound 2 was synthesized by a method similar to Example 1 using intermediate 2 and (2-methyl-4-sulfamoylphenyl)boronic acid, wherein only one boronic acid addition was required in step (i) of the method. The crude product was purified by preparative HPLC (HPLC method A) to provide compound 2 (48.5 mg, 0.09 mmol, 34%) as a white foam. LC / MS (method C): m / z 522 [M+H] + 2.28 minutes.

[0558] 1 H NMR (400MHz, DMSO-d6) δ7.94 (d, J=8.0Hz, 1H), 7.71 (s, 1H), 7.67 (m, 3H), 7.47 (br s, 2H), 7.39 (dd, J=7.6Hz, 1H), 7.36-7.27 (m, 3H), 7.23 (m, 1H), 7.17 (m, 2H), 4.83-4.74 (m, 1H), 3.84 (d, J=13.2Hz,

[0559] 1H), 3.52 (d, J=13.1Hz, 1H), 3.08 (dd, J=7.8Hz, 1H), 2.95-2.88 (m, 1H), 2.35-2.26 (m, 1H), 2.23 (s, 3H), 2.07-1.96 (m, 1H), 1.71-1.57 (m, 3H), 1.24 (d, J=6.9Hz, 3H).

[0560] No exchangeable acid protons were observed in the spectrum.

[0561] Example 3: 4-((S)-1-((R)-1-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 3)

[0562] Compound 3 was synthesized by a method similar to Example 1 using intermediate 2 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide, wherein only one boronate addition was required in step (i) of the method. The crude product was purified by HPLC (HPLC Method C) to give Compound 3 (71 mg, 0.15 mmol, 43%) as a white foam. LC / MS (Method C): m / z 486 [M+H] + 2.36 minutes.

[0563] 1 H NMR (400MHz, DMSO-d6) δ8.06-7.96 (m, 2H), 7.80-7.74 (m, 3H), 7.72 (m, 1H), 7.3 8(m, 1H), 7.33-7.18(m, 7H), 4.85-4.76(m, 1H), 3.82(d, J=13.0Hz, 1H), 3.52(d , J=13.0Hz, 1H), 3.07(dd, J=9.8, 4.1Hz, 1H), 2.98-2.90(m, 1H), 2.35-2.27(m, 1H), 2.21 (s, 3H), 2.06-1.96 (m, 1H), 1.73-1.59 (m, 3H), 1.25 (d, J=7.0Hz, 3H).

[0564] No exchangeable acid protons were observed in the spectrum.

[0565] Example 4: 4-((S)-1-((2R,4R)-4-hydroxy-1-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 4)

[0566] Compound 4 was synthesized by a method similar to Example 1 using intermediate 4 and (2-methyl-4-sulfamoylphenyl)boronic acid, wherein only one boronate addition was required in step (i) of the method. The crude residue was purified by HPLC (HPLC Method C) to give Compound 4 (73.1 mg, 0.14 mmol, 60%) as a white foam. LC / MS (Method C): m / z 538 [M+H] + 2.12 minutes.

[0567] 1 H NMR (400MHz, DMSO-d6) δ7.86 (d, J=8.0Hz, 2H), 7.81 (s, 1H), 7.75 (m, 1H), 7.46-7.34 (m, 4H ), 7.32 (d, J=8.0Hz, 2H), 7.25 (m, 1H), 4.75-5.00 (m, 1H), 4.32-4.25 (m, 1H), 3.87 (d, J=12. 9Hz, 1H), 3.69 (d, J=12.9Hz, 1H), 3.23 (dd, J=10.6, 4.6Hz, 1H), 3.10 (d, J=10.3Hz, 1H), 2. 65-2.58 (m, 1H), 2.53-2.43 (m, 1H), 2.31 (s, 3H), 1.82-1.74 (m, 1H), 1.31 (d, J=7.0Hz, 3H).

[0568] No exchangeable protons were observed in the spectrum.

[0569] Example 5: 4-((S)-1-((2R,4R)-4-hydroxy-1-((4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 5)

[0570] Compound 5 was synthesized by a method similar to Example 1 using intermediate 4 and (4-hydroxy-2-methylphenyl)boronic acid, wherein only one boronate addition was required in step (i) of the method. The crude residue was purified by HPLC (HPLC Method B) to give Compound 5 (63.6 mg, 0.13 mmol, 65%) as a white solid. LC / MS (Method C): m / z 475 [M+H] + 2.11 minutes.

[0571] 1 H NMR (400MHz, DMSO-d6) δ9.72 (br s, 1H),

[0572] 8.73(brs, 1H), 7.78(m, 2H), 7.43(s, 1H), 7.38-7.28(m, 2H), 7.21-7.16(m, 1H), 7.14-7.02 (m, 2H), 6.84 (d, J=8.3Hz, 1H), 6.65-6.62 (m, 1H), 6.58 (dd, J=8.2, 2.5Hz, 1H), 5.36 (brs, 1 H), 4.83-4.74 (m, 1H), 4.51-4.37 (m, 2H), 4.32-4.15 (m, 2H), 3.63-3.52 (m, 1H), 3.50-3.16 (m, 2H, below the water peak), 2.75-2.63 (m, 1H), 2.02 (s, 3H), 1.96-1.85 (m, 1H), 1.30 (d, J = 7.0 Hz, 3H).

[0573] Example 6: 4-((S)-1-((2R,4R)-1-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-4-hydroxypyrrolidine-2-amido)ethyl)benzoic acid (Compound 6)

[0574] Compound 6 was synthesized by a method similar to Example 1 using intermediate 4 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide, wherein only one boronate addition was required in step (i) of the method. The crude residue was purified by HPLC (HPLC Method B) to give Compound 6 (106.5 mg, 0.21 mmol, 86%) as a white foam. LC / MS (Method C): m / z 502 [M+H] + 2.12 minutes.

[0575] 1H NMR (400MHz, DMSO-d6) δ9.74 (brs, 1H), 8.74 (d, J=7.7Hz, 1H), 7.92 (s, 1H), 7.79-7.74 (m, 3H), 7.68 (dd, J= 7.8, 1.8Hz, 1H), 7.52 (s, 1H), 7.47-7.43 (m, 1H), 7.38 (m, 1H), 7.33 (s, 1H), 7.25 (dt, J=7.6, 1.5Hz, 1H), 7.0 7(m,3H),5.36(brs,1H),4.82-4.74(m,1H),4.52-4.41(m,2H),4.33-4.25(m,1H),4.25-4.18(m,1H),3.65 -3.54 (m, 1H), 3.43-3.30 (m, 1H), 2.76-2.64 (m, 1H), 2.11 (s, 3H), 1.97-1.89 (m, 1H), 1.30 (d, J=7.0Hz, 3H).

[0576] Example 7: 4-((S)-1-((R)-1-((4'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 7)

[0577] Compound 7 was synthesized by a method similar to Example 1 using Intermediate 2 and 4-hydroxyphenylboronic acid. The crude residue was purified by HPLC (HPLC Method C) to give Compound 7 (49.2 mg, 0.11 mmol, 56%) as a white foam. LC / MS (Method D): m / z 445 [M+H] + 1.82 minutes.

[0578] 1 H NMR (400MHz, DMSO-d6) δ8.07 (d, J=8.2Hz, 1H), 7.84-7.79 (m, 2H), 7.51-7.49 (m, 1H), 7.48 -7.42(m, 3H), 7.36-7.29(m, 3H), 7.25-7.20(m, 1H), 6.84-6.79(m, 2H), 4.87-4.79(m, 1H) , 3.79 (d, J = 128Hz, 1H), 3.54 (d, J = 12.8Hz, 1H), 3.09 (dd, J = 9.7, 4.4Hz, 1H), 3.01-2.95 (m , 1H), 2.36 (dd, J=8.2Hz, 1H), 2.08-1.98 (m, 1H), 1.74-1.62 (m, 3H), 1.27 (d, J=7.0Hz, 3H).

[0579] No exchangeable acid protons and hydroxyl protons were observed in the spectrum.

[0580] Example 8: 4-((S)-1-((R)-1-((2'-cyano-4'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 8)

[0581] Compound 8 was synthesized by a method similar to Example 1 using intermediate 2 and 5-hydroxy-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile, wherein only one boronate addition was required in step (i) of the method. The crude residue was purified by HPLC (HPLC Method C) to give Compound 8 (22 mg, 0.05 mmol, 32%) as a white foam. LC / MS (Method C): m / z 470 [M+H] + 1.60 minutes.

[0582] 1 H NMR (400MHz, DMSO-d6) δ8.04 (d, J=8.2Hz, 1H), 7.82-7.76 (m, 2H), 7.48 (brs, 1H), 7.45-7.3 3(m, 4H), 7.28-7.23(m, 2H), 7.22(d, J=2.6Hz, 1H), 7.17(dd, J=8.6, 2.6Hz, 1H), 4.89-4.80( m, 1H), 3.85 (d, J=13.0Hz, 1H), 3.53 (d, J=13.0Hz, 1H), 3.10 (dd, J=9.7, 4.3Hz, 1H), 3.02-2 .94 (m, 1H), 2.37-2.29 (m, 1H), 2.09-1.98 (m, 1H), 1.72-1.61 (m, 3H), 1.28 (d, J=6.9Hz, 3H).

[0583] No exchangeable acid protons and hydroxyl protons were observed in the spectrum.

[0584] Example 9: 3'-(((R)-2-(((S)-1-(4-carboxyphenyl)ethyl)carbamoyl)pyrrolidin-1-yl)methyl)-2-methyl-[1,1'-biphenyl]-4-carboxylic acid (Compound 9)

[0585] Compound 9 was synthesized by a method similar to Example 1 using intermediate 2 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid, wherein only one boronate addition was required in step (i) of the method. The crude residue was purified by HPLC (HPLC Method B) to give Compound 9 (34.7 mg, 0.07 mmol, 34%) as a white foam. LC / MS (Method C): m / z 487 [M+H] + , 1.29 minutes.

[0586] 1 H NMR (400MHz, DMSO-d6) δ12.88 (brs, 1H), 9.67 (brs, 1H), 8.93 (d, J=7.7Hz, 1H), 7.84-7.82 (m, 1H ), 7.81-7.77(m, 2H), 7.77-7.73(m, 1H), 7.48-7.41(m, 3H), 7.36-7.30(m, 1H), 7.17-7.10(m, 3H ), 4.88-4.79(m, 1H), 4.45-4.31(m, 2H), 4.21-4.13(m, 1H), 3.64-3.56(m, 1H), 3.35-3.23(m, 1H) ), 2.55-2.51(m, 1H), 2.16(s, 3H), 2.13-2.04(m, 1H), 1.96-1.81(m, 2H), 1.31(d, J=6.9Hz, 3H).

[0587] Example 10: 4-((S)-1-((R)-1-((2'-methyl-4'-(sulfamoyloxy)-1',2',3',4',5',6'-hexahydro-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 10)

[0588]

[0589] To a solution of compound 1 (74.0 mg, 0.16 mmol) in DMA (1.4 mL) was added sulfonyl chloride (46.61 mg, 0.40 mmol), and the mixture was stirred at room temperature for 18 hours, and checked for completion by LC / MS. Further sulfonyl chloride (46.61 mg, 0.40 mmol) was added, and the reaction mixture was stirred for another 18 hours, and 0.5 mL of water was subsequently added to quench the reaction. The mixture was purified by reverse phase HPLC (HPLC method B) to obtain compound 10 (68.4 mg, 0.127 mmol, 79% yield) as a colorless glass. LC / MS (method C): m / z 538 [M+H]+ , 1.88 minutes.

[0590] 1 H NMR (400MHz, MeOD-d4) δ7.87-7.83(m, 2H), 7.42-7.37(m, 3H), 7.30-7.26(m, 1H), 7.20-7.17 (m, 1H), 7.17-7.09 (m, 3H), 7.00-6.96 (m, 1H), 4.91 (q, J=7.0Hz, 1H), 4.50 (d, J=12.8Hz, 1H) , 4.29 (d, J = 12.8Hz, 1H), 4.22 (dd, J = 9.4, 6.1Hz, 1H), 3.82-3.74 (m, 1H), 3.41-3.35 (m, 1H), 2.69-2.57(m, 1H), 2.28-2.18(m, 1H), 2.14(s, 3H), 2.12-1.99(m, 2H), 1.40(d, J=7.0Hz, 3H).

[0591] No exchangeable protons were observed.

[0592] Example 11: 4-((S)-1-((R)-1-((4'-carbamoyl-2'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 11)

[0593]

[0594] Step (i): A mixture of 4-bromo-3-hydroxybenzamide (204.79 mg, 0.95 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (200.00 mg, 0.86 mmol), potassium carbonate (238.21 mg, 1.72 mmol) and tetrakis(triphenylphosphine)palladium(0) (99.58 mg, 0.09 mmol) in 1,4-dioxane (4.5 mL) and water (1 mL) was heated to 100° C. in a microwave reactor for 40 minutes. The mixture was diluted with EtOAc and 1M HCl (aq). The organic layer was separated, dried over MgSO, filtered and concentrated. The viscous residue was suspended in EtO and sonicated. The liquid was removed to give 4-(3-formylphenyl)-3-hydroxy-benzamide (150 mg, 0.6218 mmol, 72% yield) as a yellow solid which was used without further purification.

[0595] Step (ii): A mixture of intermediate 1 (150.0 mg, 0.480 mmol) and 4-(3-formylphenyl)-3-hydroxybenzamide (138.83 mg, 0.58 mmol) in DMF (3 mL) was stirred at room temperature for 30 minutes, and then sodium triacetoxyborohydride (203.27 mg, 0.96 mmol) was added. The mixture was stirred at room temperature overnight before it was directly purified by HPLC (HPLC Method A) to give methyl 4-[(1S)-1-[[(2R)-1-[[3-(4-carbamoyl-2-hydroxy-phenyl)phenyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (56.8 mg, 0.11 mmol, 23% yield) as a clear gum. LC / MS (Method C): m / z 502 [M+H] + 3.69 minutes.

[0596] Step (iii): Proceed similarly to step (ii) of Example 1 to provide Compound 11 (40.2 mg 73%) as a white foam. LC / MS (Method C): m / z 488 [M+H] + 1.72 minutes.

[0597] 1H NMR (400MHz, DMSO-d6) 812.88br(s, 1H), 9.87(s, 1H), 9.73(brs, 1H), 8.94(d, J=7.8Hz, 1H), 7.88(s, 1 H), 7.84 (d, J=8.0Hz, 2H), 7.68 (s, 1H), 7.65-7.59 (m, 1H), 7.46 (s, 1H), 7.40-7.35 (m, 3H), 7.31 (brs, 1H), 7.25 (d, J=7.9Hz, 1H), 7.18 (d, J=8.0Hz, 2H), 4.87 (q, J=7.1Hz, 1H), 4.43-4.32 (m, 2H), 4.22-4.1 4(m, 1H), 3.36-3.26(m, 1H), 3.16(s, 1H), 2.16-2.05(m, 1H), 1.96-1.82(m, 2H), 1.34(d, J=6.9Hz, 3H).

[0598] LC / MS (Method C): m / z 488 [M+H] + 1.72 minutes.

[0599] Example 12: 4-((S)-1-((R)-1-((4'-carbamoyl-6-hydroxy-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 12)

[0600]

[0601] Step (i): A mixture of intermediate 5 (100.0 mg, 0.21 mmol), (4-carbamoylphenyl)boronic acid (38.Mg, 0.23 mmol), potassium carbonate (58.15 mg, 0.42 mmol) and tetrakis(triphenylphosphine)palladium(0) (24.31 mg, 0.02 mmol) in 1,4-dioxane (1.6 mL) and water (0.50 mL) was heated to 100 °C in a microwave reactor for 40 minutes, after which time the mixture was diluted with EtOAc and water. The organic layer was separated and the aqueous layer was washed with brine, dried over MgSO4, filtered and concentrated. The residue was purified by flash column chromatography (normal phase, 10 g SNAP KP-Sil, 30 mL / min, gradient: 0% to 8% MeOH in DCM) to give methyl 4-[(1S)-1-[[(2R)-1-[[3-(4-carbamoylphenyl)-4-methoxy-phenyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (48 mg, 0.093 mmol, 44% yield) as a clear glass. LC / MS (Method B): m / z 516 [M+H] + 2.11 minutes.

[0602] Step (ii): To a solution of methyl 4-[(1S)-1-[[(2R)-1-[[3-(4-carbamoylphenyl)-4-methoxy-phenyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (45.0 mg, 0.09 mmol) in DCM (1.5 mL) was added a 1 M solution of boron tribromide in DCM (0.44 mL, 0.44 mmol) dropwise at -78°C. The mixture was gradually warmed to room temperature and stirred at room temperature overnight. The mixture was cooled to 0°C and 1:1 MeOH / H2O (2 mL) was added dropwise. The mixture was stirred for 1 hour and then concentrated, and the crude material was purified by reverse phase HPLC (HPLC Method C) to give Compound 12 (25 mg, 0.05 mmol, 59% yield) as a white foam. LC / MS (Method C): m / z 488 [M+H] + 1.64 minutes.

[0603] 1H NMR (400 MHz, methanol-d4) δ 7.97-7.87 (m, 2H), 7.87-7.79 (m, 2H), 7.62-7.49 (m, 2H), 7.41 (d, J = 2.3 Hz, 1H), 7.26 (dd, J = 8.5, 2.3 Hz, 1H), 7.13 (d, J = 8.2 Hz, 2H), 6.91 (d, J = 8.3 Hz, 1H), 4.90 (q, J = 8.5, 7.1Hz, 1H), 4.43 (d, J=12.9Hz, 1H), 4.23-4.11 (m, 2H), 3.76 (dt, J=11.6, 6.0Hz, 1H), 341 -3.33 (m, 1H), 2.64-2.50 (m, 1H), 2.31-2.14 (m, 1H), 2.12-1.94 (m, 2H), 1.40 (d, J=7.0Hz, 3H).

[0604] There are no exchangeable protons.

[0605] Example 13: 4-((S)-1-((R)-1-((4'-carbamoyl-5-hydroxy-2'-methyl-1,2,3,4,5,6-hexahydro-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 13)

[0606]

[0607] Step (i): A mixture of methyl 4-[(1S)-1-[[(2R)-1-[(3-bromo-5-methoxy-phenyl)methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate intermediate 6 (250 mg, 0.53 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (151.0 mg, 0.58 mmol), potassium carbonate (145.4 mg, 1.05 mmol) and tetrakis(triphenylphosphine)palladium(0) (60.8 mg, 0.050 mmol) in 1,4-dioxane (3.5 mL) and water (0.8 mL) was heated to 100 °C in a Biotage microwave reactor for 1 hour, after which time the mixture was diluted with EtOAc and water. The organics were separated and the aqueous layer was washed with brine, dried over MgSO4, filtered and concentrated. The residue was purified by flash column chromatography (normal phase, 10 g, The product was purified by SNAP KP-Sil-50 μm irregular silica, 30 mL / min, [Gradient: 0% to 8% MeOH in DCM]) and the resulting impure gum was sonicated in diethyl ether and the solid filtered off to give methyl 4-[(1S)-1-[[(2R)-1-[[(3-(4-carbamoyl-2-methyl-phenyl)-5-methoxy-phenyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (174 mg, 0.33 mmol, 62% yield) as a cream solid. LC / MS (Method B): m / z 530 [M+H] + 2.16 minutes.

[0608] Step (ii): To a solution of methyl 4-[(1S)-1-[[(2R)-1-[[3-(4-carbamoyl-2-methyl-phenyl)-5-methoxy-phenyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (170.mg, 0.32mmol) in DCM (5mL) was added dropwise boron tribromide, a 1M solution in DCM (1.6mL, 1.6mmol) at -78°C. The mixture was gradually warmed to room temperature and stirred at room temperature overnight. The mixture was cooled to 0°C and 1:1 MeOH / H2O (2mL) was added dropwise. The mixture was stirred for 1 hour, after which time it was concentrated. The crude material was purified by reverse phase HPLC (HPLC Method C) to give Compound 13 (89mg, 0.18mmol, 55% yield) as a white foam. LC / MS (Method C): m / z 502[M+H] + 1.66 minutes.

[0609] 1 H NMR (400 MHz, methanol-d4) δ 8.00-7.85 (m, 2H), 7.73 (s, 1H), 7.63 (d, J = 8.0 Hz, 1H), 7.19 (d, J = 8.1 Hz, 2H), 7.00 (d, J = 7.9 Hz, 1H), 6.95-6.69 (m, 3H), 4.96 (q, J = 6.7 Hz, 1H), 4.43 (d, J=12.8Hz, 1H), 4.28-4.12 (m, 2H), 3.88-3.71 (m, 1H), 340-3.35 (m, 1H), 2.67-2. 55 (m, 1H), 2.31-2.18 (m, 1H), 2.16 (s, 3H), 2.13-1.97 (m, 2H), 1.42 (d, J=7.0Hz, 3H).

[0610] There are no exchangeable protons.

[0611] Example 14: 4-((1S)-1-((2R)-1-((4'-carbamoyl-6-hydroxy-2'-methyl-1,2,3,4,5,6-hexahydro-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 14)

[0612] Compound 14 was synthesized by a method similar to Example 12 using Intermediate 5 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide to give Compound 14. LC / MS (Method C): m / z 502 [M+H] + 3.69 minutes.

[0613] 1 H NMR (400 MHz, methanol-d4) 87.94 (d, J = 8.2 Hz, 2H), 7.73 (d, J = 1.8 Hz, 1H), 7.68-7.54 (m, 1H), 7.35-7.26 (m, 1H), 7.21 (d, J = 8.1 Hz, 2H), 7.16 (d, J = 2.3 Hz, 1H), 7.09-6.96 (m, 1H), 6.90 (d, J = 8.3 Hz, 1H), 4.96 (q, J=7.0, 6.5Hz, 1H), 4.38 (d, J=12.8Hz, 1H), 4.23-4.11 (m, 2H), 3.78-3.66 (m, 1H), 3.34-3.31 (m , 1H), 2.64-2.52(m, 1H), 2.29-2.15(m, 1H), 2.11(s, 3H), 2.09-1.98(m, 2H), 1.44(d, J=7.0Hz, 3H).

[0614] There are no exchangeable protons.

[0615] Example 15: (R)-4-((1-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)methyl)-2-hydroxybenzoic acid (Compound 15)

[0616]

[0617] Step (i): Intermediate 8 (0.20 g, 0.45 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (0.15 g, 0.58 mmol) and KCO (0.123 g, 0.89 mmol) were dissolved in a mixture of 6 mL of dioxane: water (3:3) and nitrogen was purged through the mixture at room temperature for 30 minutes. PdCl(dppf)DCM (0.036 g, 0.045 mmol) was then added and the reaction mixture was stirred at 80 °C for 4 hours. The reaction mixture was then partitioned between water (70 mL) and EtOAc (70 mL). The aqueous layer was further extracted with EtOAc (2×50 mL) and the organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica). The product was eluted at 0% to 55% MeCN in water to give pure (R)-methyl 4-((1-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)methyl)-2-hydroxybenzoate (0.147 g, 65%) as a light brown solid. LC / MS: (Method A): m / z 502 [M+H] + In 1.46 minutes.

[0618] Step (ii): Methyl 4-((S)-1-((3R,6R)-6-methyl-4-(4-(trifluoromethyl)benzyl)morpholine-3-amido)ethyl)benzoate (0.147 g, 0.29 mmol) was dissolved in dioxane (2 mL) and water (1 mL). LiOH monohydrate (0.036 g, 0.88 mmol) was added at room temperature and stirred at room temperature for 16 hours. The reaction mixture was then acidified with 4N aqueous HCl (2 mL) to adjust the pH to about 1 and extracted with EtOAc (40 mL). The aqueous layer was further extracted with EtOAc (3×30 mL) and the organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 26% MeCN in water to give compound 15 as an off-white solid (0.10 g, 70%). LC / MS: (Method A): m / z 488 [M+H] + In 1.34 minutes.

[0619] 1H NMR: (400MHz, DMSO)1.79(s,3H),2.24(s,3H),3.17(s,3H),3.81(s,1H),4 .08 (s, 1H), 4.26-4.14 (m, 2H), 6.62-6.60 (d, 1H, J=8.0Hz), 6.66 (s, 1H), 7. 34-7.24(m, 3H), 7.44-7.401(m, 3H), 7.64-7.62((d, 1H, J=8.0Hz), 7.75-7 .73(d, 1H, J=7.6Hz), 7.80(s, 1H), 7.97(s, 1H), 8.53(s, 1H), 12.97(s, 2H).

[0620] Example 16: (R)-2-Hydroxy-4-((1-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)methyl)benzoic acid (Compound 16)

[0621] Compound 16 was synthesized by a method similar to Example 15 using Intermediate 8 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide to give Compound 16. LC / MS: (Method A) m / z 524 [M+H] + At 1.40 minutes.

[0622] 1 H NMR: (400MHz, DMSO): 1.77-1.74 (t, 3H, J=6.6Hz), 2.17 (s, 1H), 2.26 (s, 3H), 3.05 (s, 1H), 3.42-3.40 (m, 1H), 3.50-3.47 (m, 1H) ), 3.68 (s, 1H), 4.00-3.97 (d, 1H, J=12.4Hz), 4.25-4.13 (m, 2H), 6.57-6.56 (d, 1H, J=8.0Hz), 6.61 (s, 1H), 7.28-7.268 (d, 1H,

[0623] J=6.8Hz), 7.44-7.35(m, 6H), 7.60-7.58(d, 1H, J=8.0Hz), 7.70-7.67(dd, 1 H, J=1.6Hz&J=4.0Hz), 7.74-7.74 (d, 1H, 1.2Hz), 8.39 (s, 1H), 13.62 (s, 2H).

[0624] Example 17: 4-((S)-1-((R)-1-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoic acid (Compound 17)

[0625] Compound 17 was synthesized by a method similar to Example 15, using intermediate 10 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide to obtain compound 17. LC / MS: (Method A): The mass of the product was determined, m / z 502 [M+H] + , at 1.16 minutes.

[0626] 1 H NMR: (400MHz, DMSO) 1.28-1.26 (d, 3H, J=7.2Hz), 1.85-1.75 (m, 3H), 2.24 (s, 4H), 3.1 6 (s, 2H), 384 (s, 1H), 4.01 (s, 1H), 4.79-4.72 (m, 1H), 6.66-6.64 (d, 1H, J=7.2Hz), 6.7 2 (s, 1H), 7.30-7.23 (dd, 2H, J=7.6Hz), 7.45-7.35. (m, 4H), 7.64-7.62 (d, 1H, J=8.0H z), 7.75-7.73 (d, 1H, J=8.0Hz), 7.80 (s, 1H), 7.98 (s, 1H), 8.32 (s, 1H), 12.97 (s, 1H).

[0627] Example 18: 2-Hydroxy-4-((S)-1-((R)-1-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 18)

[0628] Compound 18 was synthesized by a method similar to Example 15 using Intermediate 10 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide to give Compound 18. LC / MS: (Method A) m / z 538 [M+H] + At 1.22 minutes.

[0629] 1H NMR: (400MHz, DMSO) 1.27-1.26 (d, 3H, J=6.8Hz), 1.84-1.75 (m, 3H), 2.26 (s, 4H), 3 .20 (s, 1H), 3.87-3.80 (bs, 1H), 4.02-3.80 (bs, 1H), 4.77-4.73 (t, 1H, J=7.2Hz), 6 .65-6.64 (d, 1H, J=6.4Hz), 6.72 (s, 1H), 7.46-7.29 (m, 7H), 7.62-7.60 (d, 1H, J=8. 0Hz), 7.69-7.670 (d, 1H, J=8.0Hz), 7.74 (s, 1H), 8.32-8.31 (m, 1H), 13.08 (s, 1H).

[0630] (Some aliphatic protons merged with the DMSO-d6 water peak).

[0631] Example 19: 4-((S)-1-((R)-1-((6-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)cyclohex-3-ene-1-carboxylic acid (Compound 19)

[0632] Compound 19 was synthesized by a method similar to Example 12 using Intermediate 5 and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide to give Compound 19. LC / MS (Method D): m / z 524 [M+H] + 1.56 minutes.

[0633] 1 H NMR (400MHz, methanol-d4) δ7.86 (d, J=8.0Hz, 2H), 7.82-7.77 (m, 2H), 7.59-7.54 (m, 2H), 7.39 (d , J=2.3Hz, 1H), 7.29-7.23 (m, 1H), 7.08 (d, J=8.0Hz, 2H), 6.89 (d, J=8.3Hz, 1H), 4.91-4.8 5 (m, 1H), 4.43 (d, J=12.9Hz, 1H), 4.22-4.07 (m, 2H), 375 (dt, J=11.6, 6.0Hz, 1H), 3.39-3. 32(m, 1H), 2.65-2.47(m, 1H), 2.27-2.13(m, 1H), 2.11-1.92(m, 2H), 1.36(d, J=7.0Hz, 3H).

[0634] There are no exchangeable protons.

[0635] Example 20: 4-((S)-1-((R)-1-((5-hydroxy-2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 20)

[0636] Compound 20 was synthesized by a method similar to Example 12 using Intermediate 6 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide to give Compound 20. LC / MS (Method D): m / z 538 [M+H] + 1.69 minutes.

[0637] 1 H NMR (400 MHz, methanol-d4) δ 7.92-7.83 (m, 2H), 7.76-7.71 (m, 1H), 7.65-7.59 (m, 1H), 7.21-7.13 (m, 2H), 7.03 (d, J = 8.1 Hz, 1H), 6.93 (dd, J = 2.3, 1.6 Hz, 1H), 6.85 (t, J = 1.5 Hz, 1H), 6.76 (dd, J = 2.4, 1.4 Hz, 1H ), 4.94 (q, J = 7.0Hz, 1H), 4.45 (d, J = 12.7Hz, 1H), 4.27-4.15 (m, 2H), 3.86-3.72 (m, 1H), 3.43-3.35 ( m, 1H), 2.69-2.55 (m, 1H), 2.31-2.17 (m, 1H), 2.17 (s, 3H), 2.15-1.98 (m, 2H), 1.41 (d, J=7.0Hz, 3H).

[0638] There are no exchangeable protons.

[0639] Example 21: 4-((S)-1-((2R,4R)-1-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-4-hydroxypyrrolidine-2-amido)ethyl)benzoic acid (Compound 21)

[0640] Compound 21 was synthesized by a method similar to Example 12 using Intermediate 6 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide to give Compound 21. LC / MS (Method D): m / z 518 [M+H] + 1.46 minutes.

[0641] 1H NMR (400MHz, methanol-d4) δ7.91-7.84(m, 2H), 7.71(dt, J=2.0, 0.6Hz, 1H), 7.64-7.59(m, 1H), 7.19-7.12(m, 2H), 6.97(d , J=8.0Hz, 1H), 6.92 (dd, J=2.3, 1.5Hz, 1H), 6.89 (t, J=1.5Hz, 1H), 6.75 (dd, J=2.3, 1.5Hz, 1H), 4.93 (q, J=7.0Hz, 1H ), 4.59-4.55 (m, 1H), 4.52 (d, J = 12.6Hz, 1H), 4.31 (dd, J = 10.8, 4.2Hz, 1H), 4.17 (d, J = 12.7Hz, 1H), 3.79 (d, J = 11.8H z, 1H), 3.43 (dd, J=11.8, 3.8Hz, 1H), 2.83 (ddd, J=14.0, 10.8, 4.7Hz, 1H), 2.19-2.08 (m, 4H), 1.42 (d, J=7.0Hz, 3H).

[0642] There are no exchangeable protons.

[0643] Example 22: 4-((1S)-1-((2R)-1-((2'-hydroxy-4'-sulfamoyl-1',2',3',4',5',6'-hexahydro-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 22)

[0644]

[0645] Step (i): Under nitrogen atmosphere, intermediate 1-HCl (0.35 g, 1.12 mmol) and 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (0.31 g, 1.34 mmol) were dissolved in DCM (4 mL). Molecular sieves (300mg) are to maintain moisture-free conditions. Potassium acetate (0.11g, 1.68mmol) is then added at room temperature, and the reaction mixture is stirred at room temperature for 4 hours. The reaction mixture is then cooled to 0 ° C, and sodium triacetoxyborohydride (0.28g, 1.34mmol) is added in batches, and the reaction mixture is stirred at room temperature for 16 hours. The solvent is removed in a vacuum, to obtain a crude product, the crude product is purified by reversed-phase gradient flash column chromatography (reverse phase, C18 silica), and the product is eluted at 0% to 22% MeCN, to obtain 4- ((S) -1- ((R) -1- (3- (4,4,5,5- tetramethyl -1,3,2- dioxaborolane -2- bases) benzyl) pyrrolidine -2- amides) ethyl) methyl benzoate (0.26g, 47.18%) as a yellow solid. LC / MS: (method A): m / z 411 [M+H] + , (boronic acid) at 1.22 minutes. & m / z 493 [M+H] + , at 1.81 minutes.

[0646] Step (ii): Methyl 4-((S)-1-((R)-1-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)pyrrolidine-2-amido)ethyl)benzoate (0.16 g, 0.32 mmol), 4-bromo-3-hydroxybenzenesulfonamide (0.10 g, 0.39 mmol) and KCO (0.13 g, 0.97 mmol) were dissolved in toluene (1 mL), ethanol (0.5 mL) and water (0.5 mL). Nitrogen was purged at room temperature for 20 minutes, and then PdCl(dppf)DCM (0.014 g, 0.016 mmol) was added and the reaction mixture was stirred at 90°C for 1.5 hours. The reaction mixture was partitioned between water (70 mL) and EtOAc (70 mL), and the aqueous layer was further extracted with EtOAc (2×40 mL). The organic layers were combined and dried (Na2SO4), the solvent was removed in vacuo, and the crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica). The product was eluted at 0% to 24% 24% MeCN in water to give methyl 4-((S)-1-((R)-1-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amide)ethyl)benzoate (0.08 g, 46.96%) as a yellow solid, which was used in the next step without further purification. LC / MS: (Method A): m / z 538 [M+H] + , at 1.33 minutes.

[0647] Step (iii): Methyl 4-((S)-1-((R)-1-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoate (0.08 g, 0.15 mmol) was dissolved in dioxane (0.5 mL) and water (0.5 mL). LiOH (0.03 g, 0.76 mmol) was added at room temperature and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated in vacuo to give the crude product, which was purified using preparative HPLC (HPLC Method D).

[0648] LC / MS: (Method A): m / z 524 [M+H] + , at 1.18 minutes.

[0649] 1 H NMR: (400MHz, CD3OD): δ1.34-1.35 (d, 3HJ=6.8Hz), 2.05-1.89 (m, 3H), 2.34 (s, 1H), 2.82-2.81 (d, 1H, J=4.8Hz), 3.38-3.37 (d, 1H, J=3.2Hz), 3.55-3.5 3(d, 1H, J=6.0Hz), 4.00(s, 2H), 7.26-7.25(d, 2H, J=7.2Hz), 7.45-7.34(m, 5H), 7.58-7.56 (d, 1H, J=7.6Hz), 7.64 (s, 1H), 7.91--7.89 (d, 2H, J=7.2Hz).

[0650] Example 23: (S)-4-(1-(2-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-2-azabicyclo[2.1.1]hexane-1-amido)ethyl)benzoic acid (Compound 23)

[0651]

[0652] Step (i): 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-1-carboxylic acid (0.25 g, 1.10 mmol) was dissolved in MeCN (3 mL), and (S)-methyl 4-(1-aminoethyl)benzoate (0.23 g, 1.32 mmol) was added to the reaction mixture at room temperature. HATU (0.63 g, 1.65 mmol) was then added, and the reaction mixture was stirred at room temperature for 30 minutes. N,N-diisopropylethylamine (0.6 mL, 3.30 mmol) was then added at room temperature, and stirred at room temperature for 2 hours. The reaction mixture was distributed between water (60 mL) and EtOAc (50 mL), and the aqueous layer was further extracted with EtOAc (2×30 mL). The organic layers were then combined and dried (Na2SO4), the solvent was removed in vacuo, and the crude product was purified by reverse phase gradient flash column chromatography (C18 silica), eluting the product at 100% MeCN to give (S)-tert-butyl 1-((1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-2-carboxylate (0.42 g, 98%) as a yellow solid. LC / MS: (Method A): m / z 289 [M+H-Boc] + At 2.23 minutes.

[0653] Step (ii): Under nitrogen atmosphere, (S)-tert-butyl 1-((1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-2-carboxylate (0.42 g, 1.08 mmol) was dissolved in dioxane (2 mL) and then 4N HCl in dioxane (4 mL) was added at room temperature and stirred for 3 hours at room temperature. The solvent was removed in vacuo and the crude material was purified by trituration with diethyl ether (5 mL) to give (S)-methyl 4-(1-(2-azabicyclo[2.1.1]hexane-1-amido)ethyl)benzoate hydrochloride (0.34 g, quantitative) as a light yellow solid. LC / MS: (Method A): m / z 289 (ES+) at 1.00 min.

[0654] Step (iii): (S)-4-(1-(2-azabicyclo[2.1.1]hexane-1-amide)ethyl)benzoic acid methyl ester hydrochloride (0.26 g, 0.80 mmol) and DIPEA (0.4 mL, 2.40 mmol) were dissolved in MeCN (3 mL) at room temperature. After this, 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (0.30 g, 0.96 mmol) was added and the reaction mixture was stirred at 80°C for 16 hours. The reaction mixture was distributed between water (80 mL) and EtOAc (80 mL), and the aqueous layer was further extracted with EtOAc (2×25 mL). The organic layers were combined and dried (Na2SO4), the solvent was removed in vacuo, and the crude product was purified by reverse phase gradient flash column chromatography (C18 silica). The product was eluted at 0% to 59% MeCN in water to give (S)-methyl 4-(1-(2-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-2-azabicyclo[2.1.1]hexane-1-amido)ethyl)benzoate (0.20 g, 48%) as a yellow solid. LC / MS (Method A): m / z 528.17 (ES+) at 1.65 min.

[0655] Step (iv): (S)-methyl 4-(1-(2-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-2-azabicyclo[2.1.1]hexane-1-amido)ethyl)benzoate (0.20 g, 0.39 mmol) was dissolved in dioxane (1 mL) and water (1 mL). LiOH monohydrate (0.084 g, 1.99 mmol) was then added at room temperature and stirred for 3 hours at room temperature. The reaction mixture was acidified with glacial acetic acid (2 mL) to adjust the pH to about 4 and then concentrated in vacuo. The crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) and the product was eluted at 0% to 36% ACN in water to give compound 23 (0.17 g, 85.19%) as an off-white solid. LC / MS (Method A): m / z 514.2 (ES+) at 1.456 min.

[0656] 1H NMR: (400MHz, DMSO) δ1.373-1.355 (d, 3H, J=7.2Hz), 1.791-1.768 (d, 4H, J=9.2Hz), 2.267 (s, 3H), 2.619-2.59 5 (d, 2H, J=9.6Hz), 2.686-2, 666 (d, 1H, J=8.0Hz), 3.475 (s, 2H), 5.041-4.986 (m, 1H), 6.594 (s, 1H), 6.848-6.8 22(d, 2H, J=10.4Hz), 7.251-7.231 (d, 1H, J=8.0Hz), 7.341 (s, 1H), 7.403-7.383 (d, 2H, J=8.0Hz), 7.735-7.71 6 (d, 1H, J=7.6Hz), 7.800-7.780 (t, 3H, J=4.0Hz), 7.968 (s, 1H), 8.228-8.208 (d, 1H, J=8.0Hz), 12.979 (s, 1H).

[0657] Example 24: 4-((S)-1-((R)-1-((5-(2-methyl-4-sulfamoylphenyl)pyridin-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 24)

[0658]

[0659] Step (i): A mixture of 5-bromonicotinaldehyde (356.81 mg, 1.92 mmol) and intermediate 1 (300.0 mg, 0.96 mmol) in DCM (4.5 mL) was stirred at room temperature for 1 hour, and then sodium triacetoxyborohydride (426.87 mg, 2.01 mmol) was added and the mixture was stirred at room temperature for 16 hours, after which it was diluted with saturated NaHCO. The organic layer was separated, washed with water and brine, dried (glass frit) and concentrated. The residue was then purified by flash column chromatography (25 g SNAP KP-Sil, 30 mL / min, gradient: 30% to 100% ethyl acetate in isohexane) to give methyl 4-[(1S)-1-[[(2R)-1-[(5-bromo-3-pyridinyl)methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (398.1 mg, 0.89 mmol, 93% yield) as a white solid. LC / MS (Method B): m / z 446 [M+H] + , 2.12 minutes.

[0660] Step (ii): A mixture of methyl 4-[(1S)-1-[[(2R)-1-[(5-bromo-3-pyridinyl)methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (200.0 mg, 0.45 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (173.11 mg, 0.58 mmol), potassium carbonate (123.86 mg, 0.90 mmol) and tetrakis(triphenylphosphine)palladium(0) (77.67 mg, 0.07 mmol) in 1,4-dioxane (1.8 mL) and water (0.45 mL) was heated to 100° C. in a microwave reactor for 30 minutes. The crude product was diluted with water and EtOAc and the organic layer was separated, washed with water, brine and dried (frit) then concentrated in vacuo. The residue was purified by flash column chromatography (50 g SNAPKP-Sil 80 mL / min, gradient: 30% to 100% ethyl acetate in isohexane) to give methyl 4-[(1S)-1-[[(2R)-1-[[5-(2-methyl-4-sulfamoyl-phenyl)-3-pyridinyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (231 mg, 96.065%) as an off-white solid. LC / MS (Method B): m / z 537 [M+H] + , 1.93 minutes.

[0661] Step (iii): To a solution of lithium hydroxide monohydrate (63.22 mg, 1.51 mmol) in water (3.2 mL) and 1,4-dioxane (3.2 mL) was added methyl 4-[(1S)-1-[[(2R)-1-[[5-(2-methyl-4-sulfamoyl-phenyl)-3-pyridinyl]methyl]pyrrolidine-2-carbonyl]amino]ethyl]benzoate (231.0 mg, 0.43 mmol) and the resulting mixture was stirred for 3 hours and then checked for completion by LC / MS. The mixture was then concentrated in vacuo and the residue was purified by reverse phase HPLC (5% to 35%, low pH) to give compound 24 (136.1 mg, 0.26 mmol, 60.5% yield) as a colorless glass. LC / MS (Method D): m / z 523 [M+H] + , 1.51 minutes.

[0662] 1H NMR (400MHz, DMSO-d6) 89.83 (br, s, 1H), δ9.01 (d, J = 7.7Hz, 1H), 8.66 (d, J = 2.0Hz, 1H), 8.53 (d, J = 2.1Hz, 1H) , 7.92 (t, J=2.1Hz, 1H), 7.83-7.78 (m, 2H), 7.77-7.73 (m, 1H), 7.71-7.64 (m, 1H), 7.36 (s, 2H), 7.22 (d, J=8.0H z, 1H), 7.15 (d, J=8.1Hz, 2H), 4.84 (p, J=7.0Hz, 1H), 4.51-4.42 (m, 2H), 4.27-4.22 (m, 1H), 3.69-3.55 (m, 1H) , 3.44-3.25 (m, 1H), 2.17 (s, 3H), 2.15-2.06 (m, 1H), 1.91 (tq, J=15.8, 8.7, 8.2Hz, 2H), 1.31 (d, J=7.0Hz, 3H).

[0663] There are no exchangeable protons.

[0664] Example 25: 4-((S)-1-((2R,4R)-1-((5-(4-carbamoyl-2-methylphenyl)pyridin-3-yl)methyl)-4-hydroxypyrrolidine-2-amido)ethyl)benzoic acid (Compound 25)

[0665] Compound 25 was synthesized by a method similar to Example 24 using Intermediate 3 and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide to give Compound 25. LC / MS (Method D): m / z 503 [M+H] + , 1.21 minutes.

[0666] 1 H NMR (400MHz, DMSO-d6) δ8.80 (d, J=7.8Hz, 1H), 8.64 (d, J=2.0Hz, 1H), 8.43 (d, J=2.1Hz, 1H), 8.00 (t, J=2.1Hz, 1H), 7.95 (s, 1H), 7.87-7.56 (m, 4H), 7.37 (s, 1H), 7.11-6.92 (m, 3 H), 4.75 (p, J=7.1Hz, 1H), 4.66-4.33 (m, 4H), 4.26 (s, 1H), 3.68-3.51 (m, 1H), 3.49-3.2 5 (m, 1H), 2.84-2.60 (m, 1H), 2.09 (s, 3H), 1.95 (d, J=13.9Hz, 1H), 1.28 (d, J=7.0Hz, 3H).

[0667] There are no exchangeable protons.

[0668] Example 26: 3-(4-Carbamoyl-2-methylphenyl)-5-(((R)-2-(((S)-1-(4-carboxyphenyl)ethyl)carbamoyl)pyrrolidin-1-yl)methyl)pyridine 1-oxide (Compound 26)

[0669]

[0670] Step (i): To a suspension of (4-carbamoyl-2-methylphenyl)boronic acid, pinacol ester (4.00 g, 15.3 mmol, 1.0 eq.), 5-bromo-3-pyridinemethanol (2.88 g, 15.3 mmol, 1 eq.) and CHCOOK (4.50 g, 45.9 mmol, 3 eq.) in dioxane / HO (64 mL / 16 mL) was added Pd(dppf)Cl (1.12 g, 1.53 mmol, 0.1 eq.) under N. The reaction mixture was stirred at 85°C for 16 hours, and then the mixture was filtered and concentrated. The residue was purified by column chromatography on silica gel (elution—DCM: MeOH=20:1→10:1) to give 4-(5-(hydroxymethyl)pyridin-3-yl)-3-methylbenzamide (2.5 g, 68%) as a yellow solid. LC / MS: (Method E): m / z 243 [M+H] + , 0.80 minutes.

[0671] Step (ii): To a solution of 4-(5-(hydroxymethyl)pyridin-3-yl)-3-methylbenzamide (1.20 g, 4.9 mmol, 1.0 equiv) in DCM (30.00 mL) was added SOCl2 (5.83 g, 49.0 mmol, 10.0 equiv). The mixture was stirred at room temperature for 1 hour, filtered and concentrated, and the residue was purified by column chromatography on silica gel (elution-DCM:MeOH=20:1→10:1) to give 4-(5-(chloromethyl)pyridin-3-yl)-3-methylbenzamide (1.0 g, 78.7%) as a yellow solid. LC / MS: (Method E): m / z 261 [M+H] + , 2.73 minutes.

[0672] Step (iii): To a solution of 4-(5-(chloromethyl)pyridin-3-yl)-3-methylbenzamide (500.0 mg, 1.92 mmol, 1.0 eq.) in DCM (15 mL) was added m-CPBA (663.5 mg, 3.84 mmol, 2.0 eq.). The reaction mixture was stirred at room temperature for 1 hour, concentrated in vacuo, and the residue was purified by column chromatography on silica gel (elution—DCM:MeOH=20:1→15:1) to give 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (500 mg, 94%) as a light yellow solid. LC / MS: (Method E): m / z 277 [M+H] + , 2.30 minutes.

[0673] Step (iv): To a solution of 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (200 mg, 0.72 mmol, 1.0 eq.) in MeCN (12 mL) and DMF (2 mL) was added Intermediate 1 (226 mg, 0.72 mmol, 1.0 eq.), NaHCO (182 mg, 2.17 mmol, 3.0 eq.) and NaI (108 mg, 0.72 mmol, 1.0 eq.). The resulting mixture was stirred at 70° C. for 4 hours, and then filtered and concentrated, and the residue was purified by column chromatography on silica gel (elution—DCM:MeOH=30:1→15:1) to give 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (311 mg, yield: 89%) as a white solid. LC / MS: (Method E): m / z 517 [M+H] + , 2.50 minutes.

[0674] Step (v): To a solution of compound 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (330 mg, 0.60 mmol, 1.0 eq.) in methanol (10 mL) was added LiOH (1.8 mL, 1.80 mmol, 3.0 eq.). The mixture was stirred at room temperature for 2 days and then diluted with water (5 mL), acidified with 1 N HCl to pH about 5, and the solvent was removed in vacuo. The residue was purified by preparative HPLC (Method E) to give compound 26 (120 mg, yield: 40.0%) as a white solid. LC / MS: (Method E): m / z 503 [M+H] + , 1.20 minutes.

[0675] 1H NMR (400 MHz, methanol-d4): 88.51-8.48 (m, 1H), 8.18 (s, 1H), 7.87 (dd, J = 8.0, 2.0 Hz, 2H), 7.80 (s, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.63-7.60 (m, 1H), 7.23 (d, J = 8.0 Hz, 2H), 6.98 (d, J = 7.6 Hz, 2H). z, 1H), 4.65-4.59 (m, 1H), 4.42-4.28 (m, 2H), 3.90 (s, 1H), 3.48-3.43 (m, 1H), 2.76-2.69 (m, 1H), 2.34-2.27 (m, 1H), 2.21-2.14 (m, 4H), 2.13-2.07 (m, 1H), 1.44 (d, J=7.2Hz, 3H).

[0676] Example 27: 4-(1-(2-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)-2-azabicyclo[2.1.1]hexane-1-amido)cyclopropyl)benzoic acid (Compound 27)

[0677] Compound 27 was synthesized by a method similar to Example 23 using methyl 4-(1-aminocyclopropyl)benzoate and intermediate 12 to give compound 27 LC / MS: (Method A): 546 [M+H] + At 1.40 minutes. Chiral HPLC: product purity was confirmed at 11.73 minutes.

[0678] 1H NMR: (400MHz, DMSO) 1.13 (m, 2H), 1.23 (m, 2H), 1.85-1.78 (m, 4H), 2.32 (s, 3H), 2.67 (s, 3H), 3.63 (s, 2H), 7.17-7. 15 (d, 2H, J=8Hz), 7.26-7.25 (d, 1H, J=6.4Hz), 7.50-7.37 (m, 6H), 7.76-7.69 (m, 4H), 8.61 (s, 1H), 12.83 (bs, 1H).

[0679] Example 28: (S)-4-(1-(2-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-2-azabicyclo[2.1.1]hexane-1-amido)ethyl)benzoic acid (Compound 28)

[0680] Compound 28 was synthesized by a method similar to Example 23 using Intermediate 13 to give Compound 28 LC / MS: (Method A): m / z 498 (ES+) at 1.27 minutes.

[0681] Chiral HPLC: product purity was confirmed at 8.83 minutes.

[0682] 1H NMR: (400MHz, DMSO): 1.39-1.37 (d, 3H, J=7.2Hz), 1.79 (m, 4H), 2.28 (s, 3H), 2.70-2.61 (m, 3H), 3.57 (s, 2H), 5.06-5. 02(t, 1H, J=7.4Hz), 7.29-7.24(m, 2H), 7.46-7.36(m, 6H), 7.82-7.75(m, 4H), 7.97(s, 1H), 8.30(s, 1H), 12.80(s, 1H).

[0683] Example 29: 4-(1-(2-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-2-azabicyclo[2.1.1]hexane-1-amido)cyclopropyl)benzoic acid (Compound 29)

[0684] Compound 29 was synthesized by a method similar to Example 23 using methyl 4-(1-aminocyclopropyl)benzoate and intermediate 13 to give compound 29 LC / MS: (Method A): m / z 510 [M+H] + At 1.23 minutes.

[0685] 1H NMR: (400MHz, DMSO).1.14(s, 2H), 1.24(s, 2H), 1.78(d, 2H, J=3.6Hz), 185(s, 2H), 2.29(s, 3H), 2.61(s, 1H), 2.67(s, 2H), 3. 62(s, 2H), 7.18-7.30(m, 4H), 7.36-7.48(m, 4H), 7.74(d, 3H, J=8.0Hz), 7.82(s, 1H), 7.98(s, 1H), 8.65(s, 1H), 12.82(s, 1H).

[0686] Example 30: (R)-4-(1-(1-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)cyclopropyl)benzoic acid (Compound 30)

[0687] Compound 30 was synthesized by a method similar to Example 23 using (tert-butoxycarbonyl)-D-proline, methyl 4-(1-aminocyclopropyl)benzoate and Intermediate 12 to give Compound 30. LC / MS: (Method A): m / z 534 [M+H] + , at 1.18 minutes.

[0688] Chiral HPLC: product purity was confirmed at 11.23 minutes.

[0689] 1H NMR: (400MHz, DMSO): 1.06-1.04 (d, 2H, J=4.8Hz), 1.19-1.18 (m, 2H), 1.78-1.77 (d, 3H, J=2. 8Hz), 2.14-2.12(m, 1H), 2.29(s, 3H), 2.38-2.36(m, 1H), 3.12-3.11(m, 2H), 3.62-3.59(d, 1H , J=12.8Hz), 3.85-3.82 (d, 1H, J=13.2Hz), 7.11-7.09 (d, 2H, J=8.4Hz), 7.28-7.27 (d, 1H, J= 6.8Hz), 7.44-7.35(m, 6H), 7.7o-7.67(m, 1H), 7.78-7.75(q, 3H), 8.48(s, 1H), 12.77(s, 1H).

[0690] Example 31: (R)-4-(1-(1-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)cyclopropyl)benzoic acid (Compound 31)

[0691] Compound 31 was synthesized by a method similar to Example 23 using (tert-butoxycarbonyl)-D-proline, methyl 4-(1-aminocyclopropyl)benzoate and Intermediate 12 to give Compound 31. LC / MS: (Method A): m / z 498 [M+H] + At 1.12 minutes.

[0692] Chiral HPLC: product purity was confirmed at 8.98 minutes.

[0693] 1H NMR: (400MHz, DMSO)1.20(m, 1H), 1.23(m, 1H), 1.77(s, 3H), 2.08-2.10(m, 1H) , 2.27(s, 3H), 2.67(s, 2H), 3.04-3.34(m, 3H), 3.60(d, 1H, J=13.2Hz), 3.82(d , 1H, J=12.8Hz), 7.06 (d, 2H, J=8.4Hz), 7.26 (d, 2H, J=7.6Hz), 7.35-7.42 (m, 4 H), 7.74 (d, 3H, J=8Hz), 7.80 (s, 1H), 7.99 (s, 1H), 8.45 (s, 1H), 12.78 (s, 1H).

[0694] Example 32: (S)-4-(1-(2-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)-2-azabicyclo[2.1.1]hexane-1-amido)ethyl)benzoic acid (Compound 32)

[0695] Compound 32 was synthesized by a method similar to Example 23 using intermediate 12 to give compound 32. LC / MS: (Method A): Product m / z 534 (ES+) at 1.27 min.

[0696] Chiral HPLC: product purity was confirmed at 12.16 minutes.

[0697] 1H NMR: (400MHz, DMSO) 1.39-1.37 (d, 3H, J=7.2Hz), 1.82-1.79 (m, 4H), 2.31 (s, 3H), 2.18-2.61 (m, 3H), 3.61-3.53 (m, 2H) , 7.26-7.24 (m, 1H), 7.47-7.37 (m, 8H), 7.72-7.70 (m, 1H), 7.80-7.76 (m, 3H), 8.28-826 (d, 1H, J=8.4Hz), 1286 (s, 1H).

[0698] Example 33: 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 33)

[0699]

[0700] Step (i): To a solution of (tert-butoxycarbonyl)-D-proline (5.0 g, 27.90 mmol) in DMF (50 mL) was added HATU (21.22 g, 55.8 mmol, 2 eq) and DIPEA (24.15 mL, 139.5 mmol, 5 eq) and the mixture was stirred at 25° C. for 0.5 h. (S)-methyl 4-(1-aminoethyl)benzoate (6.01 g, 27.90 mmol, 1 eq) was added and the resulting mixture was stirred at 25° C. for 1 h. After completion, the reaction mixture was diluted with H2O (500 mL) and extracted with EA (2×150 mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography, eluting with a gradient of 0% to 50% ethyl acetate in hexanes to give (R)-tert-butyl 2-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate (8.5 g, 79.31%) as a yellow oil.

[0701] TLC: (5.0:5.0, EA / hexene, RF: 0.7).

[0702] Mass (ESI+ve): 277.0 [M-100].

[0703] LCMS: 98.91 % (LCMS method H), RT: 2.949 min, 254 nm.

[0704] 1H NMR: (400MHz, DMSO): 1.20-1.25(m, 9H), 1.34-1.47(m, 3H), 1.78-2.13(m, 4H), 3.27-3.35(m, 2H), 3.83(m, 3H), 4.06-4.15 (m, 1H), 4.93-5.00 (m, 1H), 7.4-7.52 (m, 2H), 7.8-7.90 (m, 2H), 8.36-8.38 (d, J=6.8Hz, 1H)

[0705]

[423] Step (ii): To a solution of (R)-tert-butyl 2-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate (8.5 g, 2.85 mmol, 1 eq) in DCM (100 mL) was added HCl in dioxane (4.0 M, 85 mL). The mixture was stirred at 15°C for 1 hour. The starting material was completely consumed and the reaction mixture was then concentrated under reduced pressure to remove the HCl / dioxane. Dioxane (30 mL) was further added to the residue and concentrated under reduced pressure to give methyl 4-((S)-1-((R)-pyrrolidine-2-amido)ethyl)benzoate (7.5 g, crude, HCl salt) as a brown solid.

[0706]

[424] TLC: (5.0:5.0, EA / hexene, RF: 0.8).

[0707]

[425] Mass (ESI+ve): 277.0[M+1].

[0708]

[426] LCMS: 89.10% (LCMS method H), and RT: 2.184 min, 230 nm.

[427]

[0710] 1H NMR: (400MHz, DMSO): 1.26-1.41(m, 6H), 1.89(s, 3H), 2.33-2.35(m, 2H), 3.63-3.90(m, 9H), 3.18(m, 3H), 4.47-3.84(m, 6H), 4.21 (m, 1H) 4.97-5.0 (m, 1H), 7.50-7.52 (d, J = 4.4Hz, 2H), 7.90-7.94 (dJ = 16.0Hz, 2H), 8.51 (s, 1H), 9.29-9.30 (d, 1H). 10.03 (s, 1H).

[0711]

[428] HPLC analysis: 99.41% 5.81 RT, 240.0 nm, 4.0 nm.

[0712]

[429] Step (iii): To a solution of methyl 4-((S)-1-((R)-pyrrolidine-2-amido)ethyl)benzoate (7.5 g, 27.14 mmol, 1 eq, HCl), 3-bromo-5-methoxybenzaldehyde (7.01 g, 32.57 mmol, 1.2 eq) in DCE (40 mL) and DMF (40 ml) was added TEA (3.76 ml, 27.14 mmol, 1 eq) and the mixture was stirred at 30° C. for 1 hour. AcOH (1.63 ml, 27.14 mmol, 1 eq) and NaBH3CN (3.41 g, 54.28 mmol, 2 eq) were added and the resulting mixture was stirred at 30° C. for 12 hours. The starting material was completely consumed and one major peak with the expected mass was detected. The mixture was then diluted with H2O (500mL), NaHCO3 solid was used to adjust pH to 9, and extracted with EA (2 × 200mL). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography, eluting with a 0% to 10% MEOH / DCM gradient, to obtain pure compound 4-((S)-1-((R)-1-(3-bromo-5-methoxybenzyl)pyrrolidine-2-amide)ethyl)methyl benzoate (5.40g, 39.76%) as a white solid.

[0713]

[430] TLC: (1.0:9.0, MeOH / DCM, RF: 0.2).

[0714]

[431] Mass (ESI+ve): 475.2[M+1].

[0715]

[432] LCMS: 95.77% (LCMS method H), RT: 3.656 min, 202 nm.

[433]

[0717] 1H NMR: (400MHz, DMSO): 6.92 (s, 1H), 7.038 (s, 1H), 7.14 (s, 1H), 7.42-7.44 (d, J=8.0 Hz, 2H), 7.89-7.91 (d, J=8.0Hz, 2H), 8.18-8.20 (d, J=8.0Hz, 1H), 1.10-1.14 (m, 4H) ,1.34-1.36(d,3H),1.65-1.73(m,3H),2.020-2.068(m,1H),2.33-2.36(m,1H),2. 72-3.10 (m, 6H), 3.47-3.50 (d, J=12.0Hz, 1H), 2.70-385 (m, 6H), 4.86-4.90 (m, 1H).

[434]

[0719] 1H NMR: (400MHz, CDCl3): 6.79 (s, 1H), 7.017 (s, 1H), 7.092 (s, 1H), 7.36-7.39 (d, J=8 .0Hz, 2H), 7.68 (s, 1H), 8.037-8.058 (d, J=8.0Hz, 2H), 1.16-1.35 (m, 6H), 1.36-1.4 6(s, 3H), 1.70-1.84(m, 3H), 2.24(m, 1H), 2.44-2.46(s, 1H), 2.64(s, 1H), 2.84-3. 14(m, 5H), 3.26(m, 1H), 3.54-3.57(m, 1H), 3.72-4.002(m, 7H), 5.056-5.10(m, 1H).

[0720]

[435] HPLC analysis: 99.59%, RT, 6.57 min, at 239.0 nm.

[0721]

[436] Step (iv): To a solution of methyl 4-((S)-1-((R)-1-(3-bromo-5-methoxybenzyl)pyrrolidine-2-amido)ethyl)benzoate (5.3 g, 11.15 mmol 1 eq), (4-hydroxy-2-methylphenyl)boronic acid (2.55 g, 16.72 mmol, 1.5 eq) in dioxane (70 mL) and H2O (15 mL) was added K2CO3 (3.078 g, 11.15 mmol, 2 eq) and Pd(dppf)Cl2 (0.816 g, 1.11 mmol, 0.1 eq). The mixture was stirred at 90°C for 12 h under N2. The starting material was completely consumed and a new spot was formed. The reaction mixture was diluted with H2O (100 mL) and extracted with EA (2×100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography, eluting with a 0% to 05% MeOH / DCM gradient to give methyl 4-((S)-1-((R)-1-((4'-hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoate (3.5 g, 66%) as a white solid.

[0722]

[437] TLC: (5.0:5.0, EA / hexene, RF: 0.2).

[0723]

[438] Mass (ESI+ve): 503.2[M+1].

[0724]

[439] LCMS: 92.39% (LCMS method H), RT: 3.296 min, 230 nm.

[440]

[0726] 1H NMR: (400MHz, DMSO): 6.62-6.70 (m, 3H), 6.82-6.85 (d, J=11.2Hz, 2H), 7.00-7.02 (d, J=8.0Hz, 1H), 7.40-7.42(d, J=8.0Hz, 2H), 7.86-7.88(d, J=8.0Hz, 2H), 8.14-8.16(d, J=8.0Hz, 1H), 1.23-1.33(m, 4H), 1.72(m, 3H), 2.05(m, 1H), 2.16(s, 3H), 2.33-2.39( m, 1H), 3.087-3.097 (m, 2H), 3.49-3.52 (m, 1H), 3.76-3.94 (m, 7H), 4.85-4.90 (m, 1H).

[441]

[0728] 1H NMR: (400MHz, CDC13): 6.75-6.85(m, 5H), 7.08-7.10(d, 1H), 7.30-7.39(m, 2H), 7.87-8.06(m, 3H), 1.30-1.48(m, 3H), 1.72-1. 89(m, 4H), 2.24(s, 4H), 2.48(m, 1H), 3.18(m, 1H), 3.34-3.35(m, 2H), 3.49-3.52(m, 1H), 3.62-3.66(m, 1H), 3.85-3.95(m, 6H).

[0729]

[442] HPLC analysis: 89.40% RT 7.31 min at 239.0 nm.

[0730]

[443] Step (v): To a solution of methyl 4-((S)-1-((R)-1-((4'-hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoate (3.7 g, 7.36 mmol, 1 eq) in DCM (100 mL) was added BBr (1.0 M solution in DCM) (40.52 ml, 5.5 eq) dropwise at -60°C. After addition, the mixture was stirred at this temperature for 30 minutes, and then the resulting mixture was stirred at 15°C for 2 hours. The starting material was completely consumed. The reaction mixture was quenched by the addition of MeOH (200 mL) at -60°C, and the mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography, eluting with a 0% to 20% MeOH / DCM gradient to give methyl 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoate (2.80 g, 77.85%) as a white solid.

[0731]

[444] TLC: (4.0:1.0:5.0EA / MEOH / DCM, RF: 0.4).

[0732]

[445] Mass (ESI+ve): 489.2[M+1].

[0733]

[446] LCMS: 84.75% (LCMS method H), RT: 2.921 min, 254.0 nm.

[447]

[0735] 1H NMR: (400MHz, DMSO): 6.57-6.58(m, 2H), 6.64(d, 1H), 6.80-6.84(m, 3H), 7.17 -7.23 (d, J=8.0Hz, 2H), 7.81-7.89 (d, J=8.0Hz, 2H), 9.01-9.028 (d, J=7.2Hz, 1 H), 1.35-1.37(m, 3H), 1.90-1.93(m, 2H), 2.05-2.15(m, 4H), 2.70(m, 1H), 3.18 (s, 4H), 3.32 (m, 1H), 3.60-3.62 (s, 1H), 4.23-4.31 (m, 3H), 4.88-4.31 (m, 1H).

[0736]

[448] Chiral HPLC: 85.25%, RT 8.23 ​​min at 234.0 nm.

[0737]

[449] Step (vi): To a solution of methyl 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoate (2.80 g, 5.73 mmol) in EtOH (10 mL) and THF (30 mL) was added LiOH.HO (2.40 g, 57.31 mmol) in HO (30 mL). The mixture was stirred at room temperature for 15 hours. RM LCMS showed that the starting material remained. LiOH.HO (2.65 g, 63.04 mmol) was then added again and the mixture was stirred at room temperature for 24 hours. The starting material was completely consumed and the reaction mixture was then concentrated under reduced pressure to remove EtOH and THF. The mixture was then diluted with H2O (20 mL), the pH was adjusted to 5 with HCl solution (2.0 M), and the mixture was concentrated under reduced pressure to give a crude product. The crude product was purified by preparative HPLC (Method F) to give 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)benzoic acid (Compound 33) (500 mg, 18.38%) as a white solid.

[0738]

[450] TLC: (1.0:9.0, methanol / DCM, RF: 0.1).

[0739]

[451] Mass (ESI+ve): 475.0 [M+H].

[0740]

[452] LCMS: 100.0% (LCMS method I), RT: 2.747 min, 304.0 nm.

[0741]

[453] LCMS: 100.0% (LCMS method H), RT: 2.080 min, 304.0 nm.

[0742]

[454] HPLC analysis: 100.0% [RT 2.85 min, at 220.0 nm].

[455]

[0744] 1H NMR: (400MHz, MeOD): 6.63-6.70 (m, 5H), 6.95-6.98 (d, J=8.0Hz, 1H), 7.288-7.307 (d, J=7.6Hz, 2H), 7.94-7.97 (d, J=8.0Hz, 2H), 1.3 5-1.39 (m, 3H), 1.87-1.97 (m, 3H), 2.17 (m, 3H), 2.34 (m, 1H), 2.79-2.813 (m, 1H), 3.40 (m, 1H), 3.53-3.55 (m, 1H), 3.86-3.95 (m, 1H).

[456]

[0746] 1H NMR: (400MHz, DMSO): 6.57-6.71 (m, 5H), 6.976-6.996 (d, J=8.0Hz, 1H), 7.368- 7.388 (d, J=8.0Hz, 2H), 7.865-7.885 (d, J=8.0Hz, 2H), 8.052-8.072 (d, J=8.0H z, 1H), 9.40 (s, 1H), 1.25-1.33 (m, 3H), 1.70-1.73 (m, 3H), 2.066 (m, 2H), 2.166 (s, 3H), 3.028 (s, 1H), 3.08-3.12 (m, 1H), 3.70-3.77 (m, 2H), 3.86-4.90 (m, 1H).

[0747]

[457] Example 34: 4-((S)-1-((R)-1-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoic acid (Compound 34)

[0748]

[0749]

[458] Step (i): To a solution of intermediate 14 (160 mg, 448.38 umol, 1 eq., HCl), intermediate 15 (144.89 mg, 538.05 umol, 1.2 eq.) in DCE (2.5 mL) and DMF (2.5 mL) was added TEA (45.37 mg, 448.38 umol, 62.41 uL, 1 eq.), and the mixture was stirred at 30° C. for 1 hour. AcOH (26.93 mg, 448.38 umol, 25.64 uL, 1 eq.) and NaBH3CN (56.35 mg, 896.75 umol, 2 eq.) were added, and the resulting mixture was stirred at 30° C. for 12 hours. LC-MS showed that intermediate 14 was completely consumed, and a major peak with the expected mass was detected. The combined mixture was then diluted with H2O (20 mL), the pH was adjusted to 9 with NaHCO3 solid, and extracted with EtOAc (30 mL*2). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (HPLC method G) to obtain compound c1 (102 mg, 174.07 umol, 38.82% yield, 97.9% purity) as a yellow solid, which was determined by HNMR.

[0750]

[459] LCMS: [M+1] + =574.2.

[0751]

[460] SFC: Rt = 1.404 min, ee = 100%

[461]

[0753] 1 HNMR: (MeOD, 400MHz) δ7.80 (s, 1H), 7.75-7.71 (m, 1H), 7.67 (d, J=8.0Hz, 1H), 7. 29 (d, J=7.6Hz, 1H), 6.98 (d, J=11.2Hz, 2H), 6.91-6.85 (m, 2H), 6.79 (s, 1H), 4.85 -4.81 (m, 1H), 4.30 (q, J=6.8, 2H), 3.86-3.71 (m, 8H), 3.27-3.17 (m, 2H), 2.61-2. 49(m, 1H), 2.30(s, 3H), 2.27-2.13(m, 1H), 1.88-1.74(m, 3H), 1.39-1.29(m, 6H).

[0754]

[462] Step (ii): At -60 ° C, to a solution of compound c1 (230 mg, 400.92 umol, 1 eq.) in DCM (8 mL) was added dropwise BBr (502.20 mg, 2.00 mmol, 193.15 uL, 5 eq.) in DCM (2 mL). After addition, the mixture was stirred at this temperature for 30 minutes, and then the resulting mixture was stirred at 15 ° C for 2 hours. LC-MS showed that compound c1 was completely consumed. At -60 ° C, the reaction mixture was quenched by adding MeOH (5 mL), and the mixture was then warmed to room temperature and concentrated under reduced pressure to obtain compound c2 (220 mg, thick) as a yellow solid.

[0755]

[463] LCMS: [M+1] + =546.2.

[0756]

[464] Step (iii): To a solution of compound c2 (220 mg, 403.21 umol, 1 eq.) in EtOH (1 mL) and THF (5 mL) was added LiOH·H2O (50.76 mg, 1.21 mmol, 3 eq.) in H2O (1 mL). The mixture was stirred at 30°C for 15 hours. LCMS showed that compound c2 remained. LiOH·H2O (50.76 mg, 1.21 mmol, 3 eq.) was added and the mixture was stirred at 30°C for 15 hours. LC-MS showed that 9.1% of compound c2 remained and a main peak with the expected mass was detected. The reaction mixture was concentrated under reduced pressure to remove EtOH and THF. The mixture was then diluted with H2O (3 mL), the pH was adjusted to 5 by HCl (2 M), and the mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (HPLC method H) to give 4-((S)-1-((R)-1-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amide)ethyl)-2-hydroxybenzoic acid (Compound 34) (111.7 mg, 214.09 umol, 53.10% yield, 99.2% purity) as a white solid, which was identified by HNMR.

[0757]

[465] LCMS (LCMS method J): 1.25 min, m / z 518.34 [MH+]

[0758]

[466] SFC: Rt = 1.499 min, ee = 100%

[467]

[0760] 1HNMR (MeOD, 400MHz) δ7.79-7.71 (m, 2H), 7.66 (dd, J=1.6, 8.0Hz, 1H), 7.09 (d, J=8.0Hz, 1H), 6.91- 6.86 (m, 1H), 6.82 (s, 1H), 6.77-6.74 (m, 1H), 6.69 (d, J = 1.2Hz, 1H), 6.58 (dd, J = 1.6, 8.0Hz, 1H), 4 .84-4.80(m, 1H), 4.28-4.17(m, 1H), 4.14-4.06(m, 1H), 3.92(dd, J=4.8, 8.4Hz, 1H), 3.67-3.56(m , 1H), 3.20-3.06 (m, 1H), 2.57-2.42 (m, 1H), 2.22 (s, 3H), 2.16-1.91 (m, 3H), 1.36 (d, J=7.2Hz, 3H).

[0761]

[468] Example 35: 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoic acid (Compound 35)

[0762]

[0763]

[469] Step (i): To a solution of (tert-butoxycarbonyl)-D-proline (7.250 g, 0.027 mol, 1.0 eq) in DMF (72 mL) was added HATU (15.50 g, 0.04 mol, 1.5 eq) to the reaction mixture and the reaction mixture was stirred at 0°C for 15 min. DIPEA (13.94 mL, 0.08 mol, 3 eq) was added to the reaction mixture at 0°C and the mixture was stirred at room temperature for 30 min. (S)-1-(4-bromo-3-methoxyphenyl)ethane-1-amine was added. HCl salt (Intermediate 16) (5.85 g, 0.027 mol, 1 eq, HCl) was added and the resulting mixture was stirred at 25°C overnight. LC-MS showed that Intermediate 16 was completely consumed and one major peak with the expected mass was detected. The reaction mixture was diluted with cold H2O (800 mL) and extracted with EA (3×300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (normal phase silica, 0% to 40.0% EA /

[0764] Hexanes) to give (R)-tert-butyl 2-(((S)-1-(4-bromo-3-methoxyphenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate (11.4 g, 98.10% yield, 96% purity) as a colorless gum.

[0765]

[470] TLC: (5.0:5.0, hexane / EtOAc, RF: 0.4).

[0766]

[471] LCMS: 96.31% (LCMS Method H), RT: 3.161 min, 254.0 nm.

[0767]

[472] Chiral HPLC: 80.05% at 4.23 RT.

[473]

[0769] 1 H NMR: (400MHz, CDC13)8: 7.47-7.39 (s, 3H), 6.91-6.73 (m, 3H), 5.04 (s, 2H), 4.34 (s, 2H), 3.96-3.86 (s, 5H), 3.46-3.3 (m, 4H), 2 .99(s, 1H), 2.91-2.89(s, 1H), 2.84-2.83(s, 7H), 2.40-2.31(m, 1H), 2.19-2.08(m, 2H), 1.89-1.63(s, 5H), 1.58-1.22(m, 25H).

[0770]

[474] Step (ii): To a solution of (R)-tert-butyl 2-(((S)-1-(4-bromo-3-methoxyphenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate (11.4 g, 0.026 mol, 1 eq) in EtOH (120 mL) were added TEA (8.08 g, 0.08 mol, 11.1 mL, 3 eq) and Pd(dppf)Cl2.CH2Cl2 (2.12 g, 0.002 mol, 0.1 eq). The mixture was stirred at 100°C for 16 hours under CO gas (125 PSI). On TLC, it showed that tert-butyl (R)-2-(((S)-1-(4-bromo-3-methoxyphenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate was completely consumed and one major peak with the expected mass was detected. The reaction mixture is filtered, and the filtrate is concentrated under reduced pressure to obtain a residue. The reaction mixture is diluted with H2O (1200mL), and extracted with EA (3 × 400mL). The combined organic layer is dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue is purified by flash silica gel chromatography (normal phase silica, under 0% to 49.0% EA / hexane) to obtain (R) -2- (((S) -1- (4- (ethoxycarbonyl) -3- methoxyphenyl) ethyl) formamyl) pyrrolidine -1- carboxylic acid tert-butyl ester (7.25g, 59.37% yield) as a colorless jelly.

[0771]

[475] TLC: (5.0:5.0, hexane / EtOAc, RF: 0.4).

[0772]

[476] Mass (ESI+ve): 321.2[M-100+1].

[0773]

[477] LCMS: 89.69% (LCMS Method H), RT: 2.999 min, 254.0 nm.

[0774]

[478] Chiral HPLC: 98.91% at 6.16 RT.

[479]

[0776] 1 H NMR: (400MHz, DMSO-D2O) δ: 8.38-8.29 (m, 1H), 7.57-7.55 (s, 1H), 7.12-6.96 (m, 2H), 4.97-4.88 (m, 1H), 4.25- 4.21 (d, 2H), 4.04-4.02 (s, 1H), 3.85-3.80 (d, 3H), 2.11 (m, 1H), 1.77 (s, 3H), 1.38 (s, 8H), 1.28-1.17 (m, 11H).

[0777]

[480] Step (iii): To a solution of (R)-tert-butyl 2-(((S)-1-(4-(ethoxycarbonyl)-3-methoxyphenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate (7.25 g, 0.017 mol, 1 eq) in dioxane (20 ml) was added HCl / dioxane (4.0 M, 120 mL). The mixture was stirred at room temperature for 1.5 hours. TLC showed that tert-butyl (R)-2-(((S)-1-(4-(ethoxycarbonyl)-3-methoxyphenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate was completely consumed and a major peak with the expected mass was detected. The reaction mixture was concentrated under reduced pressure to remove HCl / dioxane, dioxane (40 ml*2) was added, and the reaction mixture was concentrated under reduced pressure to remove HCl / dioxane to give ethyl 2-methoxy-4-((S)-1-((R)-pyrrolidine-2-amido)ethyl)benzoate.HCL salt (5.5 g, crude) as a brown gum.

[0778]

[481] TLC: (5.0:5.0, hexane / EtOAc, RF: 0.2).

[0779]

[482] LCMS: 98.97% (LCMS method K), RT: 1.057 min, 254.0 nm.

[0780]

[483] Chiral HPLC: 97.97% at 6.19 RT.

[484]

[0782] 1 H NMR: (400MHz, DMSO-D20) δ: 9.79 (s, 1H), 9.23-9.21 (d, 1H), 8.54 (s, 1H), 7.61-7.59 (d, 1H), 7.16 (s, 1H), 7.01-6.99 (d, 1H), 4.99-4.95 (m, 1H), 4. 27-4.21(q, 3H), 3.85(s, 3H), 3.58(s, 2H), 3.22-3.18(m, 3H), 2.70(s, 3H ), 2.37-2.35(m, 1H), 1.91(s, 3H), 1.42-1.40(d, 3H), 1.26-1.32(t, 4H).

[0783]

[485] Step (iv): A mixture of ethyl 2-methoxy-4-((S)-1-((R)-pyrrolidine-2-amido)ethyl)benzoate.HCL salt (5.50 g, 0.015 mol, 1 eq.) was dissolved in DCE (50 ml), and TEA (4.66 g, 6.14 ml, 0.046 mol, 3 eq.) was added at 0°C. The mixture was stirred at 0°C for 10 minutes, and then 4'-hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-carbaldehyde (Intermediate 17) (3.73 g, 0.015 mol, 1 eq.) was added to the reaction mixture, and the mixture was stirred at 60°C for 3 hours. After 3 hours, NaBH3CN (2.86 g, 0.046 mol, 3 eq.) was added, and the resulting mixture was then stirred at 30°C overnight. LC-MS showed that ethyl 2-methoxy-4-((S)-1-((R)-pyrrolidine-2-amido)ethyl)benzoate.HCL salt was completely consumed, and a major peak with the expected mass was detected. The mixture was then diluted with H2O (600 mL) and extracted with EA (400 mL*3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by normal phase column chromatography (normal phase neutral activated alumina, at 5.0% to 30.0% methanol / DCM) to give ethyl 4-((S)-1-((R)-1-((4'-hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-methoxybenzoate (3.77 g, 44.72% yield, 81.66% purity) as an off-white solid.

[0784]

[486] TLC: (7.0:3.0, hexane / EtOAc, RF: 0.4).

[0785]

[487] Mass (ESI+ve): 547.33 (M+1).

[0786]

[488] LCMS: 81.66% (LCMS method H), RT: 3.382 min, 254 nm.

[0787]

[489] Chiral HPLC: 83.20%, RT 8.08 min at 232.0 nm.

[490]

[0789] 1H NMR: (400MHz, DMSO-D2O) δ: 1.21-1.1.32(m, 6H), 1.72-1.75(m, 3H), 2.01-2.06(m, 1H), 2.18(s, 3H ), 2.35-2.39(m, 1H), 3.05-3.13(m, 2H), 3.52-3.56(m, 1H), 3.79-3.81(m, 6H), 4.21-4.26(m, 2H), 4.83-4.86 (m, 1H), 6.64-6.69 (m, 2H), 6.73 (s, 1H), 6.84-6.87 (d, 2H, J=10Hz), 6.91-6.93 (d, 1H, J =7.6Hz), 7.02-7.07 (m, 2H), 7.55-7.57 (d, 1H, J = 7.6Hz), 8.14-8.16 (d, 1H, J = 7.6Hz), 9.37 (s, 1H).

[0790]

[491] Step (v): To a solution of ethyl 4-((S)-1-((R)-1-((4'-hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-methoxybenzoate (3.77 g, 0.006 mol, 1 eq) in DCM (108 mL) at -78°C was added BBr (1.0 M solution in DCM) (8.63 g, 34.42 ml, 0.034 mol, 5 eq) to the reaction mixture and the mixture was stirred at -78°C for 1 hour. The resulting mixture was then stirred at room temperature for 2 hours. On TLC, ethyl 4-((S)-1-((R)-1-((4'-hydroxy-5-methoxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-methoxybenzoate was completely consumed, and one major peak with the expected mass was detected. The reaction mixture was quenched by slowly adding MeOH (197.5 mL) dropwise at -78°C, and then the mixture was warmed to room temperature and concentrated under reduced pressure to give ethyl 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoate (5.04 g, crude) as a yellow solid.

[0791]

[492] TLC: (8.0:2.0, hexane / EtOAc, RF: 0.2).

[0792]

[493] Mass (ESI+ve): 519.4[M+1].

[0793]

[494] LCMS: 52.0% (LCMS method L), RT: 2.838 min, 254 nm.

[0794]

[495] Step (vi): To a solution of ethyl 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoate (5.04 g, 0.009 mol, 1 eq) in EtOH (35 mL) and THF (170 mL) was added LiOH.H2O (3.05 g, 0.07 mol, 7.5 eq) in H2O (35 mL). The mixture was stirred at 30°C for 7 days. LC-MS showed that ethyl 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoate was completely consumed and a major peak with the expected mass was detected. The reaction mixture was concentrated under reduced pressure to remove EtOH and THF. The mixture was then cooled at 0°C, diluted with H2O (20 mL), and filtered through HCl at 0°C.

[0795] The mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (HPLC Method I) to give 4-((S)-1-((R)-1-((4',5-dihydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)pyrrolidine-2-amido)ethyl)-2-hydroxybenzoic acid (Compound 35) (472.5 mg, 9.91% yield, 100.0% purity) as a white solid.

[0796]

[496] TLC: (1.0:9.0, methanol / DCM, RF: 0.1).

[0797]

[497] Mass (ESI+ve): -491.2 (M+1).

[0798]

[498] LCMS: 100.0% (LCMS method L), RT: 2.415 min, 304.0 nm.

[0799]

[499] LCMS: 99.60% (LCMS Method H), RT: 2.156 min, 304.0 nm.

[0800]

[500] Chiral HPLC: 100.0% at 6.75 RT.

[501]

[0802] 1H NMR: (400MHz, MeOD) δ: 7.76-7.78 (d, 1H), 6.86-6.88 (d, 1H), 6.79-6.80 (d, 2H), 6.72 (s, 2H), 6.66 (s, 1H), 6.60 (s, 2H), 4.85-4.87 (m, 1H) , 4.18-4.21(d, 1H), 4.08-4.11(d, 1H), 3.94(s, 1H), 3.60(s, 1H), 3.15(s, 1H), 2.49(s, 1H), 2.12(s, 4H), 1.99(s, 2H), 1.37-1.39(d, 3H).

[502]

[0804] 1 H NMR: (400MHz, DMSO-D2O) δ: 9.52 (s, 1H), 935 (s, 1H), 8.14-8.31 (m, 1H), 7.64-7.66 (d, 1H), 6.93-6.95 (d, 1H), 6.72 (s, 3H), 6.11-6.65 (m, 4H), 4.76-4.79(m, 1H), 3.90(s, 1H), 3.72(s, 1H), 3.2-3.5(m, 3H), 2.33-2.36(s, 1H), 2.12(s, 3H), 1.75-1.85(m, 3H), 1.29-1.28(d, 4H).

[0805] Biological activity assay

[0806]

[503] Cloning, baculovirus production, large-scale infection and membrane preparation of HEK293 cells: Human prostaglandin E2 receptor 4 (EP4) was cloned into the pBacMam expression vector (GeneScript, UK). Transposition of EP4 DNA was performed using the Bac-to-Bac baculovirus expression system of Invitrogen. P0 baculovirus was generated by transfecting SF9 cells with bacmid DNA using Cellfectin II transfection reagent (ThermoFisherScientific, UK). After the P0 generation, P1 virus was then generated for large-scale infection and membrane preparation. HEK293 cells were cultured in DMEM (ThermoFisher Scientific, UK) supplemented with 10% heat-inactivated fetal bovine serum (FBS). The cells were incubated at 500 cm 3Infected with 3.5 million cells / mL inoculation density under 5% v / v EP4Bacman in the bottle. At 37°C and 5% CO2, expression was performed within a 36-hour period. Cells were removed using PBS and a cell scraper. Cell culture was centrifuged at 4°C for 10 minutes at 2500RPM. The supernatant was then poured out, and the precipitate was stored at -80°C. The precipitate was thawed and resuspended in 15mL homogenizing buffer (20mM HEPES, 10mM EDTA, pH 7.4). Homogenized in a mechanical homogenizer (VMR) for 10 seconds. The membrane was centrifuged at 4°C for 15 minutes at 40,000g in a centrifuge tube. The supernatant was poured out and resuspended in 15mL homogenizing buffer and homogenized for 20 seconds. The membrane was centrifuged at 4°C for 45 minutes at 40,000g. The membrane was then resuspended in 3 mL of well-mixed storage buffer (20 mM HEPES, 0.1 mM EDTA, pH 7.4). The resulting membrane was then stored at -80°C.

[0807]

[504] cAMP Gs functional assay: cAMP production following EP4 receptor activation was determined using the Homogeneous Time-Resolved Fluorescence (HTRF) cAMP dynamic-2 assay (Cisbio, France). HEK293 cells were transfected with 0.5% EP4 Bacmam virus for 36 hours, then cells were dissociated and frozen at -150°C.

[0808]

[505] On the day of the assay, increasing concentrations of test compounds were added to ProxiPlate-384 Plus, white 384-shallow well microplates (PerkinElmer, USA) using an ECHO dispenser along with a positive control (10 μM PGE2 (Tocris, Abingdon, UK)) and a negative control (DMSO (Sigma-Aldrich, UK)).

[0809]

[506] The cells were thawed in a water bath and resuspended in DMEM supplemented with 10% FBS, then centrifuged at 1200 RPM for 5 minutes to form a pellet. The pellet was resuspended in assay buffer (DMEM + 0.5 mM IBMX (Tocris, Abingdon, UK)) to a concentration of 0.5 × 10 6Cells / mL. The cell suspension was added to the pre-assigned assay plate using multidrop, with a final assay concentration of 5000 cells / well. The plate was then incubated at 37°C and 5% CO2 for 30 minutes. cAMP production was determined according to the manufacturer's instructions, and the plate was then read on a PheraStar fluorescence plate reader (BMG LabTech, Germany).

[0810]

[507] Dotmatics was used to calculate the pEC based on the midpoint of the curve. 50 The values ​​are shown in Table 2.

[0811]

[508] Table 2: EP4pEC 50 and Emax value

[0812]

[0813] One-way CACO-2 cell permeability assay

[0814]

[509] Caco-2 cells (ECACC) were cultured at 2×10 5 Cells were seeded into 24-well Transwell plates and used for confluent monolayers after culturing for 21 days at 37°C, 5% CO2. Test compounds were incubated in assay buffer (Hanks balanced salt solution supplemented with 25 mM HEPES, adjusted to pH 6.5) at 10 μM, 0.2% DMSO final concentration, n=2. Hanks balanced salt solution supplemented with 25 mM HEPES (0.2% DMSO final concentration) adjusted to pH 7.4 was used for the basolateral chamber (as a receiver).

[0815]

[510] Incubations were performed at 37°C with samples removed from both the donor and acceptor compartments at T = 0 and 1 hour and compounds analyzed by mass spectrometry (LC-MS / MS) containing an analytical internal standard (0.5 μM carbamazepine).

[0816]

[511] Apparent permeability (P app ) values ​​are shown in Table 3 and are determined according to the following relationship:

[512]

[0818]

[0819]

[513] Where V is the volume of each Transwell compartment (top 125 μL, basolateral 600 μL), and Concentration is the relative MS response of compound in the donor compartment before incubation and in the receptor compartment at the end of incubation (normalized to internal standard). Area = area of ​​cells exposed for drug transfer (0.33 cm 2 ).

[0820]

[514] Lucifer Yellow (LY) was added to the top buffer in all wells to assess the viability of the cell layer. Since LY cannot freely penetrate the lipophilic barrier, a high degree of LY transport indicates poor integrity of the cell layer and rejection of LY. app >10×10 -6 cm / sec hole.

[0821]

[515] Compounds recovered from the wells were determined based on the MS responses in the donor and acceptor compartments at the end of the incubation period compared to the response in the donor compartment before the incubation period (normalized to an internal standard).

[0822]

[516] Table 3 - Average apparent permeability (P app )value

[0823]

[0824]

[0825] Numbered implementation plan

[0826] 1. A compound of formula I or a pharmaceutically acceptable salt or tautomer thereof:

[0827]

[0828] in:

[0829] A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R', S(O)2R', S(O)2OR', SO2N(R')2, C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0830] Ring B is aryl or heteroaryl;

[0831] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0832] R 1 and R 2 Each independently is H, C 1-6 Alkyl, C 1-6 Alkoxy, or R 1 and R2 Together with the carbon atom to which they are attached, they form C 3-6 Cycloalkane-1,1-diyl ring;

[0833] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0834] R 4 It is C 1-6 Alkyl, halogen, CN, NO2 or OR';

[0835] R 5 Is H or C 1-6 Alkyl; or R 4 and R 5 Together with the pyrrolidine ring to which they are attached, they form C 1-6 Alkylene linker;

[0836] R 6 It is H, C 1-6 Alkyl, halogen, CN, NO2, OR', CO2R' or C(O)R';

[0837] R 7 Is OR', OC(O)R', OC(O)OR', CO2R', CON(R')2, SO2N(R')2, SO2R', OSO2R' or OSO2N(R')2;

[0838] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0839] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0840] n and m are each independently 0, 1, 2 or 3;

[0841] wherein at each occurrence, alkyl, alkylene and cycloalkyl are each optionally and independently substituted by OH, SH, CN, NO2, COOH, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0842] wherein at each occurrence, heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO2, CO2R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0843] 2. A compound according to embodiment 1, wherein ring B is a 5- to 6-membered aryl or a 5- to 6-membered heteroaryl; each of which is optionally and independently substituted by OR', SR', CN, NO2, CO2R', halogen or C 1-4 Up to 3 instances of substitution in the alkyl group.

[0844] 3. A compound according to embodiment 1, wherein ring B is phenyl optionally substituted with up to three instances of OH or is a 6-membered heteroaryl containing one or two nitrogen atoms, each of which is optionally oxidized.

[0845] 4. A compound of formula I, which is a compound of formula (1) or a pharmaceutically acceptable salt thereof:

[0846]

[0847] in:

[0848] A is selected from:

[0849]

[0850] U, V, W and Z are each independently selected from CH, COH, N or N + -O - , wherein at least three of U, V, W and Z are CH;

[0851] X and Y are each independently selected from CH, CF, COH or N;

[0852] R 1 and R 2 are independently H, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkyl, or R 1 With R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring;

[0853] R 3 is H, OH or F;

[0854] R 4 is H, OH, F or R 4 With R 5 connected to form a CH2 bridge;

[0855] R 5 Is H or R 5 With R 4 connected to form a CH2 bridge;

[0856] R 6 is H, OH, CN, halogen, C optionally substituted by 1 to 3 fluorine atoms 1-3alkoxy, or C optionally substituted by 1 to 3 fluorine atoms 1-3 alkyl;

[0857] R 7 is OH, CO2H, CONH2, SO2NH2 or OSO2NH2; and

[0858] R 9 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring.

[0859] 5. A compound according to any one of embodiments 1 to 4, wherein R 5 It's H.

[0860] 6. A compound according to any one of embodiments 1 to 5, wherein A is:

[0861]

[0862] 7. The compound according to embodiment 4, which is a compound of formula (2a) or a pharmaceutically acceptable salt thereof:

[0863]

[0864] Among them U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , R 6 and R 7 Same as defined in Embodiment 4.

[0865] 8. A compound according to any one of embodiments 1 to 7, wherein R 1 is H or methyl, or R 1 With R 2 Connect to form a cyclopropane-1,1-diyl ring.

[0866] 9. The compound according to embodiment 8, wherein R 1 It's methyl.

[0867] 10. A compound according to any one of embodiments 1 to 9, wherein R 2 It's H.

[0868] 11. The compound according to embodiment 4, which is a compound of formula (3a) or a pharmaceutically acceptable salt thereof:

[0869]

[0870] Among them U, V, W, X, Y, Z, R 3 , R 4, R 6 and R 7 Same as defined in Embodiment 4.

[0871] 12. A compound according to any one of embodiments 1 to 11, wherein R 3 It's H.

[0872] 13. A compound according to any one of embodiments 1 to 12, wherein R 4 It's H.

[0873] 14. A compound according to any one of embodiments 1 to 13, wherein R 6 It is H, OH, CN or methyl.

[0874] 15. The compound according to embodiment 14, wherein R 6 It's methyl.

[0875] 16. A compound according to any one of embodiments 1 to 15, wherein R 7 It is CONH2 or SO2NH2.

[0876] 17. A compound according to any one of embodiments 1 to 16, wherein both X and Y are CH.

[0877] 18. A compound according to any one of embodiments 4 to 17, wherein U, V, W and Z are CH, or wherein U, V and Z are CH and W is COH.

[0878] 19. The compound or pharmaceutically acceptable salt according to embodiment 1, wherein the compound is selected from:

[0879]

[0880]

[0881]

[0882] or a pharmaceutically acceptable salt thereof.

[0883] 20. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 19, and a pharmaceutically acceptable excipient.

[0884] 21. A compound according to any one of embodiments 1 to 19 or a composition according to embodiment 20 for use in the treatment of an EP4 receptor mediated disease.

[0885] 22. The compound or composition for use according to embodiment 21, wherein the EP4 receptor-mediated disease is a gastrointestinal disorder.

[0886] 23. The compound or composition for use according to embodiment 22, wherein the gastrointestinal disorder is selected from constipation disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastric and intestinal damage, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrheal diseases, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease and ischemic colitis.

[0887] 24. The compound or composition for use according to embodiment 21, wherein the EP4 receptor-mediated disease is a pulmonary disease or disorder.

[0888] 25. The compound or composition for use according to embodiment 24, wherein the pulmonary disease or disorder is selected from chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperresponsive airway disorder and idiopathic pulmonary fibrosis.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof: in: A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R', S(O)2R', S(O)2OR', SO2N(R')2, C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl; Ring B is aryl or heteroaryl; X and Y are each independently CR" or N, wherein at least one of X and Y is CH; R 1 and R 2 Each independently is H, C 1-6 Alkyl, C 1-6 Alkoxy, or R 1 and R 2 Together with the carbon atom to which they are attached, they form C 3-6 Cycloalkane-1,1-diyl ring; Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl; R 4 It is H, C 1-6 Alkyl, halogen, CN, NO2 or OR'; R 5 Is H or C 1-6 Alkyl; or R 4 and R 5 Together with the pyrrolidine ring to which they are attached, they form C 1-6 Alkylene linker; R 6 It is H, C 1-6 alkyl, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkoxy, halogen, CN, NO2, OR', CO2R' or C(O)R'; R 7 is OR’, OC(O)R’, OC(O)OR’, CO2R’, CON(R’)2, SO2N(R’)2, SO2R’, OSO2R’ or OSO2N(R’)2; Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl; Each R" is H, C 1-6 alkyl, halogen or OR'; and n and m are each independently 0, 1, 2 or 3; wherein at each occurrence, alkyl, alkylene and cycloalkyl are each optionally and independently substituted by OH, SH, CN, NO2, COOH, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group; wherein at each occurrence, heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO2, CO2R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

2. The compound according to claim 1, wherein Ring B is a 5- to 6-membered aryl or a 5- to 6-membered heteroaryl; each of which is optionally and independently substituted by OR', SR', CN, NO2, CO2R', halogen or C 1-4 Up to 3 instances of substitution in the alkyl group.

3. The compound according to claim 1, wherein Ring B is phenyl optionally substituted with up to three instances of OH or is a 6-membered heteroaryl containing one or two nitrogen atoms, each of which is optionally oxidized.

4. The compound according to any one of claims 1 to 3, which is a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof: in: U, V, W and Z are each independently selected from CH, COH, N or N + -O - , wherein at least three of U, V, W and Z are CH.

5. A compound according to any one of claims 1 to 4, wherein R 4 is H, OH, F, or R 4 With R 5 connected to form a CH2 bridge; preferably wherein R 4 is H, OH or R 4 With R 5 Connect to form a CH2 bridge.

6. A compound according to any one of claims 1 to 5, wherein R 5 Is H or R 5 With R 4 connected to form a CH2 bridge; preferably wherein R 5 It's H.

7. The compound according to any one of claims 1 to 6, wherein A is selected from: Where R 9 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring; preferably wherein A is:

8. The compound according to claim 4, which is a compound of formula (2a) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof: Among them U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , R 6 and R 7 Same as defined in claim 4.

9. A compound according to any one of claims 1 to 8, wherein R 1 and R 2 are independently H, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkyl, or R 1 With R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 Cycloalkyl ring.

10. A compound according to any one of claims 1 to 9, wherein R 1 is H or methyl, or R 1 With R 2 connected to form a cyclopropane-1,1-diyl ring; more preferably wherein R 1 is methyl; and / or wherein R 2 It's H.

11. The compound according to claim 4, which is a compound of formula (3a) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof: Among them U, V, W, X, Y, Z, R 3 , R 4 , R 6 and R 7 Same as defined in claim 4.

12. A compound according to any one of claims 1 to 11, wherein R 3 is H, OH or F; preferably wherein R 3 is H or OH; more preferably wherein R 3 It's H.

13. A compound according to any one of claims 1 to 12, wherein R 4 is H, OH or F; preferably wherein R 4 is H or OH; more preferably wherein R 4 It's H.

14. A compound according to any one of claims 1 to 13, wherein R 6 is H, OH, CN, halogen, C optionally substituted by 1 to 3 fluorine atoms 1-3 alkoxy, or C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkyl; preferably wherein R 6 is H, OH, CN or methyl; more preferably wherein R 6 It's methyl.

15. A compound according to any one of claims 1 to 14, wherein R 7 is OH, CO2H, CONH2, SO2NH2 or OSO2NH2; preferably wherein R 7 It is CONH2 or SO2NH2.

16. A compound according to any one of claims 1 to 15, wherein X and Y are each independently selected from CH, CF, COH or N; preferably wherein X is CH and Y is CH or COH; more preferably wherein both X and Y are CH.

17. A compound according to any one of claims 4 to 16, wherein U, V, W and Z are CH, or wherein U, V and Z are CH and W is COH.

18. The compound according to claim 1, wherein the compound is selected from: or a pharmaceutically acceptable salt, solvate, hydrate or tautomer thereof.

19. A pharmaceutical composition comprising the compound, pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer according to any one of claims 1 to 18, and a pharmaceutically acceptable excipient.

20. The pharmaceutical composition of claim 19, wherein the composition further comprises at least one additional therapeutic agent selected from the group consisting of aminosalicylate, corticosteroid, immunomodulator, and combinations thereof.

21. A kit comprising a compound, pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer according to any one of claims 1 to 18, and at least one additional therapeutic agent selected from the group consisting of aminosalicylate, corticosteroid, immunomodulator and combinations thereof.

22. A compound according to any one of claims 1 to 18, a composition according to claim 19 or 20, or a kit according to claim 21 for use in medicine.

23. A compound according to any one of claims 1 to 18, a composition according to claim 19 or 20, or a kit according to claim 21 for use in the treatment of an EP4 receptor mediated disease.

24. The compound, composition or kit for use according to claim 23, wherein the EP4 receptor-mediated disease is a gastrointestinal disorder.

25. A compound, composition or kit for use according to claim 24, wherein the gastrointestinal disorder is selected from constipation disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastric and intestinal damage, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrheal diseases, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease and ischemic colitis.

26. The compound, composition or kit for use according to claim 23, wherein the EP4 receptor mediated disease is a pulmonary disease or disorder.

27. A compound, composition or kit for use according to claim 26, wherein the pulmonary disease or disorder is selected from chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperresponsive airway disorder and idiopathic pulmonary fibrosis.

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