Morpholine-3-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] This application relates to new compounds and their use as prostaglandin E 2 prostaglandin E receptor 4 2 receptor 4, EP 4 The compounds described herein can be used to treat or prevent diseases involving EP 4 The present application also relates to pharmaceutical compositions comprising these compounds, and to the use of these compounds and compositions in preventing or treating diseases involving EP 4 The preparation and use of receptors in such diseases. Background Art

[0002] Prostaglandins (including prostaglandins and thromboxanes) are metabolic derivatives of arachidonic acid that play a key role in cellular physiological functions. Arachidonic acid is catabolized by phospholipase A2 (PLA 2 The biosynthesis of prostaglandins is mediated by cyclooxygenase (COX), which catalyzes the conversion of arachidonic acid into an unstable intermediate (PGH 2 ) and lead to PGE 2 The production of prostaglandins. 2 Represents the most widely produced prostaglandins, whose activity is mediated through action on four functionally distinct receptor subtypes, EP1-4.

[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 signal transduction 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 colon innervating sensory neurons, and may provide benefits in suppressing inappropriate mucosal immune responses and protecting the GI mucosal barrier. PGE2 signaling through EP4 promotes cell differentiation into a wound-associated epithelial cell phenotype, 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 through 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 a prostaglandin E 2 Receptor 4 (EP 4 ) compounds that have agonist activity.

[0014] Provided herein are compounds of Formula I or pharmaceutically acceptable salts, solvates, hydrates, tautomers or optical isomers thereof:

[0015]

[0016] in:

[0017] A is OR', C(O)R', CO 2 R', C(O)N(R') 2 ,C(O)N(R')S(O) 2 R',S(O) 2 R',S(O) 2 OR', SO 2 N(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, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkoxy, halogen, CN, NO 2 、OR'、CO 2 R' or C(O)R';

[0023] R 5 is OR', OC(O)R', OC(O)OR', CO 2 R', CON(R') 2, SO 2 N(R') 2 , SO 2 R', OSO 2 R' or OSO 2 N(R') 2 ;

[0024] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0025] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0026] n is 0, 1, 2, or 3;

[0027] wherein at each occurrence, alkyl and cycloalkyl are each optionally and independently replaced by OH, SH, CN, NO 2 , CO 2 H, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0028] wherein heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO 2 , CO 2 R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0029] In another aspect, the present invention includes a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable excipient.

[0030] 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.

[0031] 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.

[0032] In another aspect, the invention includes a compound described herein, a composition described herein, or a kit described herein for use in treating an EP4 receptor mediated disease.

[0033] 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.

[0034] 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.

[0035] The compounds herein can be used as EP 4 Receptor agonists. The compounds herein can selectively act on EP 4 The compound can be used to prepare a composition or a drug. The compound, composition or drug can be used to treat, prevent, improve, control, or involve EP 4 Diseases or conditions involving EP receptors or decreased 4 The compound, composition or drug can be used to treat, prevent, improve, control or reduce the risk of gastrointestinal disorders and disorders, 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.

[0036] 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

[0037] The present invention relates to novel compounds. The present invention also relates to novel compounds as EP 4 The present invention also relates to the use of new compounds in the preparation of EP 4 Receptor agonists for use in medicine, and including administration as EP 4 Receptor agonists of the present invention are used in therapeutic methods.

[0038] The compounds of formula I can be used to treat, prevent, improve, control 4 Diseases or conditions involving EP receptors or decreased 4The compounds of formula I can be used to treat, prevent, improve, control or reduce the risk of gastrointestinal disorders and disorders, 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.

[0039] 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.

[0040] Certain novel compounds of the present invention are shown to be EP 4 Particularly high activity of receptor agonists.

[0041] The compounds of the present invention have been shown to have the potential to act as EP 4 The compounds of the invention also have low gastrointestinal permeability as shown by Caco-2 studies. Therefore, it is believed that the compounds of the invention exhibit low systemic bioavailability when administered orally. 4 Functional agonists of the receptor have the potential to treat a range of gastrointestinal disorders. 4 The combination of receptor agonist activity and low gastrointestinal permeability suggests that the compounds of the invention may be useful in treating a range of gastrointestinal disorders without the cardiovascular side effects that would result from systemic distribution.

[0042] In one aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0043]

[0044] in:

[0045] A is OR', C(O)R', CO 2 R', C(O)N(R') 2 ,C(O)N(R')S(O) 2 R',S(O) 2 R',S(O) 2OR', SO 2 N(R') 2 , C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0046] Ring B is aryl or heteroaryl;

[0047] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0048] 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;

[0049] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0050] R 4 It is H, C 1-6 alkyl, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkoxy, halogen, CN, NO 2 、OR'、CO 2 R' or C(O)R';

[0051] R 5 is OR', C(O)R', CO 2 R', C(O)N(R') 2 ,C(O)N(R')S(O) 2 R',S(O) 2 R',S(O) 2 OR', OSO 2 N(R') 2 ;

[0052] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0053] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0054] n is 0, 1, 2, or 3;

[0055] wherein at each occurrence, alkyl and cycloalkyl are each optionally and independently replaced by OH, SH, CN, NO 2 , CO2 H, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0056] wherein heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO 2 , CO 2 R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0057] In some embodiments, the compound is a compound of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0058]

[0059] in:

[0060] A is OR', C(O)R', CO 2 R', C(O)N(R') 2 ,C(O)N(R')S(O) 2 R',S(O) 2 R',S(O)3OR',SO 2 N(R') 2 , C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0061] Ring B is aryl or heteroaryl;

[0062] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0063] 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;

[0064] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0065] R 4 It is H, C 1-6 Alkyl, halogen, CN, NO 2 、OR'、CO 2 R' or C(O)R';

[0066] R 5 is OR', OC(O)R', OC(O)OR', CO 2 R', CON(R') 2 , SO 2 N(R') 2 , SO 2 R', OSO 2 R' or OSO 2 N(R') 2 ;

[0067] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0068] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0069] n is 0, 1, 2, or 3;

[0070] wherein at each occurrence, alkyl and cycloalkyl are each optionally and independently replaced by OH, SH, CN, NO 2 , CO 2 H, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0071] wherein heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO 2 , CO 2 R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0072] In some embodiments, the compound is a compound of Formula I, (1), (1a), (1b), (2), (2a), (2b), (3), (3a), (3b), (4), (4a), (4b), IIa, IIb, IIc, IId or IIe, or a pharmaceutically acceptable salt or tautomer thereof.

[0073] In some embodiments, the compound is a compound of Formula I, (1), (1a), (1b), (2), (2a), (2b), (3), (3a), (3b), (4), (4a), (4b), IIa, IIb, IIc, IId or IIe, or a pharmaceutically acceptable salt thereof.

[0074] In some embodiments, provided herein are compounds of Formula IIa, IIb, IIc, IId or IIe, or pharmaceutically acceptable salts, solvates, hydrates, tautomers or optical isomers thereof:

[0075]

[0076] Among them, A, X, Y, R 1 , R 2 , R 3 , R 4 and R 5 As defined above.

[0077] In some embodiments, A is CO 2 R'、C(O)N(R')S(O) 2 R', S(O) 2 R' or heteroaryl.

[0078] In some embodiments, A is COOH or a 5-membered heteroaryl, which is optionally substituted with OR' or SR'.

[0079] 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, NO 2 , CO 2 R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0080] In some embodiments, Ring B is a 5- to 6-membered aryl or a 5- to 6-membered heteroaryl.

[0081] In some embodiments, Ring B is phenyl optionally substituted with up to three instances of OH.

[0082] In some embodiments, Ring B is a 6-membered heteroaryl containing one or two nitrogen atoms, and each nitrogen is optionally substituted with oxo.

[0083] In some embodiments, Ring B is a phenyl, pyridine, or pyridine-N-oxide ring, each of which is optionally substituted with OR'.

[0084] In some embodiments, each R 3 are independently selected from OR', halogen or C 1-6 alkyl.

[0085] In some embodiments, each R 3 are independently selected from OR' or alkyl; and n is 1.

[0086] In some embodiments, n is 0, 1, or 2.

[0087] In some embodiments, n is 0 or 1.

[0088] In some embodiments, n is 0.

[0089] In some embodiments, R 4 It is H, C 1-6 Alkyl, halogen or OR'.

[0090] In some embodiments, R 4 It is H, methyl or OH.

[0091] In some embodiments, R 5 is OR', OC(O)R', CO 2 R', CON(R') 2 、SO 2 N(R') 2 、SO 2 R' or OSO 2 N(R') 2 .

[0092] In some embodiments, R 5 Is OR', CO 2 R', CON(R') 2 、SO 2 N(R') 2 or OSO 2 N(R') 2 .

[0093] In some embodiments, R 5 OH, CO 2 H.CONH 2 、SO 2 NH 2 or OSO 2 NH 2 .

[0094] In some embodiments, R 5 It is CONH 2 or SO 2 NH 2 .

[0095] In some embodiments, R 5 It is CONH 2 .

[0096] In some embodiments, the compound is a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0097]

[0098] wherein 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; X, Y, R 1 , R2 , R 3 , R 4 and R 5 As described in this article.

[0099] In some embodiments, the compound is a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0100]

[0101] in:

[0102] A is selected from:

[0103]

[0104] 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; and

[0105] X and Y are each independently selected from CH, CF, COH or N;

[0106] 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;

[0107] R 3 is H, OH or F;

[0108] R 4 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;

[0109] R 5 OH, CO 2 H.CONH 2 、SO 2 NH 2 or OSO 2 NH 2 ;and

[0110] R 6 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring.

[0111] In some embodiments, A is selected from:

[0112]

[0113] And R 6 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring.

[0114] In some embodiments, A is:

[0115]

[0116] In some embodiments, R 3 is halogen or OR'. In another embodiment, R 3 is F or OH, and n is 0, 1 or 2.

[0117] In some embodiments, X and Y are each independently selected from CH, CF, COH, or N.

[0118] In some embodiments, at least one of X and Y is CH.

[0119] In some embodiments, X and Y are both CH.

[0120] In some embodiments, provided herein are compounds of Formula IIa, Formula IIb, Formula IIc, Formula IId, or Formula IIe.

[0121] In some embodiments, R 1 and R 2 Each independently is H, C 1-6 Alkyl, halogen or hydroxyl, or R 1 and R 2 Together with the carbon atom to which they are attached, they form C 3-6 Cycloalkyl.

[0122] In some embodiments, R 1 and R 2 are each independently selected from 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.

[0123] In some embodiments, R 1 and R 2 are each independently H or methyl, or R 1 and R 2 Together with the carbon atom to which they are attached they form a cyclopropyl group.

[0124] In some embodiments, R 4It is H, OH, halogen, C 1-6 Alkoxy or C 1-6 In another embodiment, R 4 It is H, OH or methyl.

[0125] In some embodiments, R 5 Is OR', CO 2 R', CON(R') 2 、SO 2 N(R') 2 or OSO 2 N(R') 2 In another embodiment, R 5 OH, CO 2 H.CONH 2 、SO 2 NH 2 or OSO 2 NH 2 .

[0126] 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.

[0127] In some embodiments, the present invention includes a pharmaceutical composition comprising a compound or salt described herein and a pharmaceutically acceptable excipient.

[0128] 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.

[0129] In another embodiment, 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 salt or composition described herein.

[0130] In another embodiment, the present invention includes a method of treating a disease mediated by the EP4 receptor, comprising administering to a patient in need thereof a compound or salt or composition described herein.

[0131] In one embodiment of this aspect, the EP4 receptor mediated disease is a gastrointestinal disorder.

[0132] In another embodiment, 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.

[0133] In some embodiments, provided herein are compounds of formula (1a) or (1b), or pharmaceutically acceptable salts, solvates, hydrates, tautomers or optical isomers thereof:

[0134]

[0135] Among them U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 and R 5 As described in this article.

[0136] In some embodiments, provided herein are compounds of formula (2), formula (2a), or formula (2b), or pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof:

[0137]

[0138] Among them A, U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 and R 5 As described in this article.

[0139] In some embodiments, provided herein are compounds of formula (2a) or (2b), or pharmaceutically acceptable salts, solvates, hydrates, or tautomers thereof:

[0140]

[0141] in:

[0142] 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; and

[0143] X and Y are each independently selected from CH, CF, COH or N;

[0144] 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;

[0145] R 3 is H, OH or F;

[0146] R 4 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

[0147] R 5 OH, CO 2 H.CONH 2 、SO 2 NH 2 or OSO 2 NH 2 .

[0148] In some embodiments, provided herein are compounds of formula (3), formula (3a) or formula (3b), or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0149]

[0150] Among them A, U, V, W, X, Y, Z, R 3 , R 4 and R 5 As described in this article.

[0151] In some embodiments, provided herein is a compound of formula (3a) or formula (3b), or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof:

[0152]

[0153] in:

[0154] 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;

[0155] X and Y are each independently selected from CH, CF, COH or N;

[0156] R 3 is H, OH or F;

[0157] R 4 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

[0158] R 5 OH, CO 2 H.CONH 2 、SO 2 NH 2 or OSO 2 NH 2 .

[0159] In some embodiments, provided herein are compounds of formula (4), formula (4a) or formula (4b), or pharmaceutically acceptable salts, solvates, hydrates or tautomers thereof:

[0160]

[0161] Among them A, U, V, W, X, Y, Z, R 3 , R 4 and R 5 As described in this article.

[0162] In some embodiments, the compound is a compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0163]

[0164] in:

[0165] A is

[0166] U is CH or N;

[0167] V is CH or N;

[0168] W is selected from CH, COH, N or N+-O-;

[0169] Z is selected from CH, COH or N;

[0170] X is CH or N;

[0171] Y is CH, CF, COH or N;

[0172] R 1 and R 2 are each independently selected from H or methyl, or R1 and R 2 Together with the carbon atom to which they are attached they form a cyclopropyl group;

[0173] R 3 is H, OH or F;

[0174] R 4 is selected from H, methyl or OH; and

[0175] R 5 Selected from OH, CONH 2 、SO 2 NH 2 or OSO 2 NH 2 .

[0176] In some embodiments, the compound is a compound of formula (1a) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof:

[0177]

[0178] in:

[0179] U is CH or N;

[0180] V is CH or N;

[0181] W is selected from CH, COH, N or N+-O-;

[0182] Z is selected from CH, COH or N;

[0183] X is CH or N;

[0184] Y is CH, CF, COH or N;

[0185] R 1 and R 2 are each independently selected from H or methyl, or R 1 and R 2 Together with the carbon atom to which they are attached they form a cyclopropyl group;

[0186] R 3 is H, OH or F;

[0187] R 4 is selected from H, methyl or OH; and

[0188] R 5 Selected from OH, CONH 2 、SO 2 NH 2 or OSO 2 NH 2 .

[0189] In some embodiments, the compound is a compound of formula (2), formula (2a), or formula (2b), and U, V, and Z may be CH, and W may be COH.

[0190] In some embodiments, A may be selected from: CO 2 H, tetrazole, 1,2,4- oxadiazol-5(2H)-one, 1,3,4- 2-oxadiazole-2(3H)-one, CONHSO 2 R 6 、CONHSO 2 Me, SO 3 H, 1, 3, 4- oxadiazole-2(3H)-thione, 1,2,4- 1,2,4-thiadiazol-5(2H)-one, 1,2,5-thiadiazolidine-3-one 1,1-dioxide and 2,4- Oxazolidinedione.

[0191] In one embodiment, A may be selected from:

[0192]

[0193] In some embodiments, A can be selected from CO 2 H and a tetrazole ring. In another embodiment, A can be CO 2 H. In another embodiment, A can be a tetrazole ring.

[0194] In some embodiments, U may be CH. In another embodiment, U may be COH. In another embodiment, U may be N. In another embodiment, U may be N + -O - .

[0195] In some embodiments, U is CH or N.

[0196] In some embodiments, V may be CH. In another embodiment, V may be COH. In another embodiment, V may be N. In another embodiment, V may be N + -O - .

[0197] In some embodiments, V is CH or N.

[0198] In some embodiments, W may be CH. In another embodiment, W may be COH. In another embodiment, W may be N. In another embodiment, W may be N +-O - .

[0199] In some embodiments, W is selected from CH, COH, N or N + -O - .

[0200] In some embodiments, Z may be CH. In another embodiment, Z may be COH. In another embodiment, Z may be N. In another embodiment, Z may be N + -O - .

[0201] In some embodiments, Z is selected from CH, COH, or N.

[0202] In some embodiments, at least three of U, V, W, and Z are CH. In another embodiment, U, V, and W may be CH. In another embodiment, U, V, and Z may be CH. In another embodiment, U, W, and Z may be CH. In another embodiment, V, W, and Z may be CH. In another embodiment, U, V, W, and Z may be CH. In another embodiment, U, V, and Z may be CH, and W may be COH.

[0203] In some embodiments, X may be CH. In another embodiment, X may be CF. In another embodiment, X may be COH. In another embodiment, X may be N.

[0204] In some embodiments, X can be CH or N.

[0205] In some embodiments, Y may be CH. In another embodiment, Y may be CF. In another embodiment, Y may be COH. In another embodiment, Y may be N.

[0206] In some embodiments, Y can be CH, CF, COH, or N.

[0207] In some embodiments, at least one of X and Y is CH. In another embodiment, both X and Y may be CH.

[0208] In some embodiments, X can be CH or N, and Y can be CH, CF, COH, or N.

[0209] In some embodiments, R 1 It may be H or C optionally substituted by 1 to 3 fluorine atoms. 1-3 In another embodiment, R 1 It may be H. In another embodiment, R 1It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 In another embodiment, R 1 Can be C 1-3 In another embodiment, R 1 Can be used with R 2 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 In another embodiment, R 1 Can be used with R 2 Connect to form C 3-6 In another embodiment, R 1 It can be H or methyl, or it can be combined with R 2 In another embodiment, R 1 It may be methyl optionally substituted by 1 to 3 fluorine atoms. 1 In another embodiment, R 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.

[0210] In some embodiments, R 2 It may be H. In another embodiment, R 2 It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 In another embodiment, R 2 Can be C 1-3 In another embodiment, R 2 Can be used with R 1 connected to form C optionally substituted by 1 to 3 fluorine atoms 3-6 In another embodiment, R 2 Can be used with R 1 Connect to form C 3-6 In another embodiment, R 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.

[0211] In some embodiments, R 3 It may be H. In another embodiment, R 3 It may be OH. In another embodiment, R 3 It can be F.

[0212] In some embodiments, R 3 It can be H, OH or F.

[0213] In some embodiments, R 3 Can be H or F.

[0214] In some embodiments, R 4 and R 5 Together with the rings they are connected to, they can be selected from:

[0215]

[0216] In some embodiments, R 4 It can be H. R 4 It may be OH. In another embodiment, R 4 Can be CN. In another embodiment, R 4 In another embodiment, R 4 It may be F. In another embodiment, R 4 Can be Cl. In another embodiment, R 4 It can be Br. In another embodiment, R 4 It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 In another embodiment, R 4 Can be C 1-3 In another embodiment, R 4 It may be a methoxy group optionally substituted by 1 to 3 fluorine atoms. 4 In another embodiment, R 4 It may be C optionally substituted by 1 to 3 fluorine atoms 1-3 In another embodiment, R 4 Can be C 1-3 In another embodiment, R 4 It can be H or methyl. In another embodiment, R 4 It may be methyl optionally substituted by 1 to 3 fluorine atoms. 4 It may be methyl.

[0217] In some embodiments, R 4 is selected from 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.

[0218] In some embodiments, R 5 It may be OH. In another embodiment, R 5 It can be CO 2 H. In another embodiment, R 5 It can be CONH 2 In another embodiment, R 5 It can be SO 2 NH 2 In another embodiment, R 5 It can be OSO 2 NH 2 .

[0219] In some embodiments, R 5 Can be OH, CONH 2 、SO 2 NH 2 or OSO 2 NH 2 .

[0220] In some embodiments, R 6 Can be C 1-3 In another embodiment, R 6 Can be C 3-6 In another embodiment, R 6 It may be methyl.

[0221] In some embodiments, the compounds or pharmaceutically acceptable salts disclosed herein have EP4 receptor agonist activity.

[0222] 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.

[0223] In some embodiments, the method of treating an EP4 receptor mediated disease is a gastrointestinal disorder.

[0224] 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.

[0225] 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.

[0226] In some embodiments, the compound of Formula I or Formula (1) is a compound listed in Table 1 or a pharmaceutically acceptable salt thereof.

[0227] Table 1 - Exemplary compounds of formula (I)

[0228]

[0229]

[0230]

[0231]

[0232]

[0233] abbreviation

[0234] aq aqueous solution

[0235] Bn Benzyl

[0236] DCM Dichloromethane

[0237] DMA Dimethylacetamide

[0238] DMF Dimethylformamide

[0239] dppf 1,1'-Bis(diphenylphosphino)ferrocene

[0240] EDCI 1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide

[0241] EtOAc Ethyl acetate

[0242] HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-Oxide Hexafluorophosphate

[0243] HOBt Hydroxybenzotriazole

[0244] HPLC High Performance Liquid Chromatography

[0245] hr hour

[0246] hrs hours

[0247] LC / MS Liquid Chromatography Mass Spectrometry

[0248] M Moore

[0249] MeCN Acetonitrile

[0250] MeOH Methanol

[0251] N equivalent (normal)

[0252] Preparative HPLC Preparative High Performance Liquid Chromatography

[0253] RT Room temperature

[0254] sat saturated

[0255] THF Tetrahydrofuran

[0256] UPLC Ultra-Performance Liquid Chromatography

[0257] definition

[0258] In this application, unless otherwise indicated, the following definitions apply.

[0259] The term "treatment" in connection with the use of any of the compounds described herein (including those of Formula (A)-(C), (1), (1a), (1b), (2), (2a), (2b), (3), (3a), (3b), (4), (4a), and (4b)) is used to describe any form of intervention in which the compound is administered to a subject suffering from, 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 manifests measurable or detectable symptoms of a disease or condition.

[0260] 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.

[0261] As used herein, the term "hydroxyl" or "hydroxy" refers to an -OH moiety.

[0262] 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, heteroarylcarbonylamino, heteroaralkylcarbonylaminoalkylaminocarbonyl, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, arylaminocarbonyl, or heteroarylaminocarbonyl], amino [e.g., aliphatic amino, alicyclic amino, or heteroalicyclic amino], sulfonyl [e.g., aliphatic -SO 2 -], sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfonamide, oxo, carboxyl, carbamoyl, alicyclic oxy, heteroalicyclic oxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, or hydroxy. 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-SO 2 -amino)alkyl), aminoalkyl, amidoalkyl, (alicyclic)alkyl, or haloalkyl.

[0263] As used herein, "alkylene" or "cycloalkane" groups refer to divalent branched alkyl, straight chain alkyl, or cyclic alkyl groups containing 2 to 12 (e.g., 2 to 8, 2 to 6, or 2 to 4) carbon atoms and are used to connect two chemical moieties. Some examples of alkylene include, but are not limited to, methylene, ethylene, propylene, butylene, isopropylene (methylethylene) and isobutylene (2-methylpropylene). Alkylene or cycloalkane may be substituted (i.e., optionally substituted) with one or more substituents as defined in the alkyl.

[0264] 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.

[0265] 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.

[0266] 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 4-8 The 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; (aromatic aliphatic) oxy; (heteroaromatic aliphatic) oxy; aroyl; heteroaroyl; amino; oxo (on the non-aromatic carbocyclic ring of the benzo-fused bicyclic or tricyclic aryl); nitro; carboxyl; amide; acyl [e.g., (aliphatic) carbonyl; (alicyclic) carbonyl; ((alicyclic) aliphatic) carbonyl; (aromatic aliphatic) carbonyl; (heteroalicyclic) carbonyl; ((heteroalicyclic) aliphatic) carbonyl; or (heteroaromatic aliphatic) carbonyl]; sulfonyl [e.g., aliphatic -SO 2 - or amino-SO 2 -]; 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.

[0267] 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.

[0268] 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.

[0269] 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, (aroliphatic)oxy, (heteroaroliphatic)oxy, aroyl, heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino, (cycloaliphatic)carbonylamino, ((cycloaliphatic)aliphatic)carbonylamino, (aryl)carbonyl alkyl)carbonyl, ((heteroaliphatic)carbonylamino, ((heteroalicyclic)aliphatic)carbonylamino, (heteroaryl)carbonylamino, or (heteroaromatic)carbonylamino], nitro, carboxyl [e.g., HOOC-, alkoxycarbonyl, or alkylcarbonyloxy], acyl [e.g., (alicyclic)carbonyl, ((alicyclic)aliphatic)carbonyl, (aromatic)carbonyl, (heteroalicyclic)carbonyl, ((heteroalicyclic)aliphatic)carbonyl, or (heteroaromatic)carbonyl], cyano, halogen, hydroxy, mercapto, sulfonyl [e.g., alkyl-SO 2 - and aryl-SO 2 -], sulfinyl [e.g., alkyl-S(O)-], sulfanyl [e.g., alkyl-S-], sulfoxy, urea, thiourea, sulfamoyl, sulfonamide, oxo, or carbamoyl.

[0270] 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.

[0271] 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.

[0272] 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.

[0273] 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, anthenyl, 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.

[0274] Heteroaryl groups include N-oxide heteroaryl compounds which are oxidized at the nitrogen atom, such as pyridine, pyrimidine, pyrazine, pyrrole, imidazole, thiazole, quinoline or isoquinoline. Examples of N-oxide heteroaryl groups are those having the formula C 5 H 5 N - →O + Pyridine-N-oxide.

[0275] 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.

[0276] 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.

[0277] 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.

[0278] 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 ...

[0279] As used herein, an "alkoxy" group refers to an alkyl-O- group, wherein "alkyl" has been previously defined.

[0280] 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-.

[0281] As used herein, a "thiol" group refers to -SH.

[0282] As used herein, a "sulfo" group when used terminally refers to -SO 3 H or -SO 3 R X , or -S(O) when used internally 3 -.

[0283] 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 -NRY -, where R X , R Y and R Z Defined above.

[0284] 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.

[0285] 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 -S(O) when used internally 2 -NR x -or-NR x -S(O) 2 -, where R x , R y and R Z Defined above.

[0286] 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.

[0287] As used herein, a "halogen" or "halo" group refers to fluorine, chlorine, bromine, or iodine.

[0288] As used herein, "oxo" refers to =0.

[0289] 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.

[0290] 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.

[0291] 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. XO(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.

[0292] As used herein, "aliphatic chain" refers to a branched or straight chain aliphatic group (eg, an alkyl, alkenyl, or alkynyl group). A straight chain aliphatic chain has the structure -[CH 2 ]v-, wherein 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.

[0293] 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 can be optionally substituted by 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 can form a ring together with the atoms to which they are bound.

[0294] 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.

[0295] 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.

[0296] To the extent that any of the compounds described possess chiral centers, the 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, however prepared.

[0297] Insofar as any compound disclosed herein has an acid or basic center such as a carboxylate / ester or an amino group, all salt forms of the compound are included herein. The invention described herein also relates to all tautomers of the compounds described herein. In the case of pharmaceutical use, the salt should be considered a pharmaceutically acceptable salt.

[0298] 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.

[0299] 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.

[0300] 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 acid (e.g., D-gluconic acid), glucuronic acid (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.

[0301] 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.

[0302] 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.

[0303] 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).

[0304] 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 and 14 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.

[0305] 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.

[0306] 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.

[0307] Combination therapy

[0308] 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.

[0309] 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.

[0310] 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.

[0311] 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.

[0312] In one embodiment, the compound of the invention and the additional therapeutic agent are administered to a subject separately, sequentially or simultaneously.

[0313] 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.

[0314] 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).

[0315] 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 separately administering the first amount of the compound of the present invention and the second amount of the additional therapeutic agent.

[0316] 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.

[0317] The presence of synergy can be determined using a suitable method for evaluating drug interactions. Suitable methods include, for example, Sigmoid-Emax equation (Holford, NHG and Scheiner, LB, Clin. Pharmacokinet. 6: 429-453 (1981)), Loewe additivity equation (Loewe, S. and Muischnek, H., Arch. Exp. Pathol Pharmacol. 114: 313-326 (1926)) and median effect equation (Chou, TC and Talalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). Each equation mentioned above can be applied together with experimental data to produce corresponding figures for helping to evaluate the effect of drug combinations. The corresponding graphics associated with the equation mentioned above are concentration-action curves, isobologram curves and combination index curves, respectively.

[0318] 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.

[0319] 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.

[0320] 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.

[0321] 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.

[0322] 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.

[0323] 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.

[0324] Pharmaceutical composition

[0325] While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (eg, formulation).

[0326] Therefore, in another embodiment of the present invention, there is provided a pharmaceutical composition comprising at least one compound of formula I as defined above and at least one pharmaceutically acceptable excipient.

[0327] 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.

[0328] The composition may be a tablet composition.

[0329] The composition may be a capsule composition.

[0330] 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.

[0331] 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.

[0332] Pharmaceutical compositions containing compounds of formula I may be formulated according to known techniques, see for example Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.

[0333] The pharmaceutical composition may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ocular, otic, rectal, intravaginal or transdermal administration.

[0334] 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.

[0335] 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.

[0336] 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.

[0337] The pharmaceutical composition generally comprises about 1% (w / w) to about 95%, preferably % (w / w) of active ingredient and 99% (w / w) to 5% (w / w) of pharmaceutically acceptable excipients (e.g., as defined above) or a combination of such excipients. Preferably, the composition comprises about 20% (w / w) to about 90% (w / w) of active ingredient and 80% (w / w) to 10% of pharmaceutical excipients or a combination of excipients. The pharmaceutical composition comprises about 1% to about 95%, preferably about 20% to about 90% of active ingredient. The pharmaceutical composition according to the present invention can be, for example, in the form of a unit dose, such as an ampoule, a vial, a suppository, a prefilled syringe, a dragee, a powder, a tablet or a capsule.

[0338] 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.

[0339] 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.

[0340] 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.

[0341] The compound of formula I is usually present in a unit dosage form, and therefore usually 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, a specific subrange of the compound is 0.1 milligram to 2 grams of active ingredient (more usually 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).

[0342] 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.

[0343] 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.

[0344] Process for preparing the compound of formula (I)

[0345] Additional embodiments are disclosed in more detail in the following examples, which are not in any way intended to limit the scope of the claims.

[0346] General synthetic scheme

[0347] Compounds of formula I can be prepared according to synthetic methods known to those skilled in the art and as described herein.

[0348] Solution 1

[0349]

[0350] In Scheme 1, compounds of formula (2a) can be prepared as outlined in Scheme 1. Compounds of formula G4 (wherein PG 1 The alkylation is carried out by reacting an amine with a suitable acid protecting group, such as a methyl ester. Typically, the alkylation is carried out in the presence of a base, such as NaHCO 3 In the presence of and in a solvent such as MeCN, an alkylation reaction is carried out to obtain a tertiary amine of formula G6. 1 The resulting ester is deprotected under conditions related to the properties of the compound (usually in a solvent such as MeOH, in the presence of a nucleophilic base such as lithium hydroxide, to hydrolyze the methyl ester functional group) to obtain an acid of formula (7). An amide bond formation reaction between an acid of formula G7 and an amine of formula G8a in a solvent such as DCM, in the presence of an amide coupling reagent (such as HOBt or EDCI) and a base (such as triethylamine) yields a compound of formula 2a.

[0351] In compounds (where A is a protected carboxylic acid group, where PG 2 represents a suitable acid protecting group such as a methyl ester), coupling produces an amide of formula G9b. Amide G9b can be used with a protecting group PG 2 The methyl ester functional group is further deprotected under conditions related to the nature of the ester. Typically, the methyl ester functional group is hydrolyzed in a solvent such as MeOH in the presence of a nucleophilic base such as lithium hydroxide to give a compound of formula 2b, see Scheme 2.

[0352] Solution 2

[0353]

[0354] Alternatively, in Scheme 3, the compound of Formula 2a can be synthesized from the compound of Formula G12, as described in Schemes 3 and 4. According to Scheme 3, the compound of Formula G12 can be provided by the following method: in a solvent such as MeCN or DMF, in the presence of a suitable coupling agent such as HATU and a base such as N,N-diisopropylethylamine, an acid of Formula G10 (wherein PG 3 Representing a suitable amine protecting group such as BOC) is coupled with an amine of formula G8a to produce the desired amide of formula G11. 3Deprotection is carried out under conditions related to the nature of the (usually BOC deprotection in a suitable solvent such as 1,4-dioxane in the presence of an acid such as HCl) to provide the desired amine intermediate of formula G12. Alternatively, the amine of formula G12 can be produced by coupling the amino acid G13 with the amine of formula G8a using amide bond forming conditions as described above.

[0355] Solution 3

[0356]

[0357] Subsequently, the amine of formula G12 can be benzylated by reductive amination with an aldehyde of formula G14 to give the corresponding tertiary amine of formula G16. Typical reductive amination conditions utilize reagents such as sodium triacetoxyborohydride or sodium cyanoborohydride in a solvent such as DCM or methanol at temperatures from room temperature to 70°C in the presence or absence of a promoter such as glacial acetic acid or zinc chloride. Alternatively, tertiary amines of formula 16 can be prepared by alkylating the amine of formula G12 with an alkylating agent of formula G15 (wherein LG represents a suitable leaving group such as a halide, typically a bromide). Typical alkylation conditions utilize a suitable base such as potassium carbonate, N,N-diisopropylethylamine or sodium bicarbonate at temperatures from room temperature to 80°C in a solvent such as acetonitrile or DMF in the presence or absence of sodium iodide. The resulting compound of formula G16 (wherein Hal is a suitable halide, typically a bromide) can be subjected to a Suzuki coupling reaction with a boronic acid or boronic ester of formula G17 (wherein R is an alkyl group, such as a methyl group) to convert it into a compound of formula 2a. The cross-coupling reaction is carried out in the presence of a catalyst (e.g., tetrakistriphenylphosphine palladium 0 or PdCl) together with a base such as potassium carbonate. 2 dppf.DCM) and typically in a solvent combination such as 1,4-dioxane and water, with the reaction temperature ranging from room temperature to 100°C, with or without microwave irradiation.

[0358] Solution 4

[0359]

[0360] In the compounds of Scheme 3 and Scheme 4 (where A is a protected carboxylic acid group, wherein PG 2 represents a suitable acid protecting group such as a methyl ester), according to Scheme 5, a compound of formula G16 is coupled with a compound of formula G17 to produce a compound of formula G9b. 2 The compound of formula G9b is deprotected under conditions related to the nature of the compound (usually in a solvent such as MeOH in the presence of a nucleophilic base such as lithium hydroxide to hydrolyze the methyl ester function) to provide the carboxylic acid compound of formula 2b.

[0361] Solution 5

[0362]

[0363] The skilled person will appreciate that the reaction steps described in Schemes 3 to 5 can be combined in different ways as needed to successfully prepare the desired compound of formula 2a. This may include additional steps, for example, functional group modification, protection and / or deprotection steps in the overall synthesis sequence. For example, the compound of formula 2a can be prepared as shown in Scheme 6. Under the conditions previously outlined in Scheme 4, the reductive amination reaction between the secondary amine of formula G12 and the aldehyde of formula G19 provides the desired tertiary amine of formula G20. Using the conditions described in Scheme 4, the subsequent Suzuki reaction with the appropriate aryl halide of formula G21 (wherein Hal is typically a bromide) provides the compound of formula 2a. As described in Scheme 2, the subsequent deprotection of the ester of formula 18 gives the compound of formula 2a.

[0364] Solution 6

[0365]

[0366] In the compound of Scheme 6 (wherein A is a protected carboxylic acid group, wherein PG 2 represents a suitable acid protecting group, such as a methyl ester), according to Scheme 7, a compound of formula G20 is coupled with a compound of formula G21 to produce a compound of formula G9b. 2 The compound of formula G9b is deprotected under conditions related to the nature of the compound (usually in a solvent such as MeOH in the presence of a nucleophilic base such as lithium hydroxide to hydrolyze the methyl ester function) to provide the carboxylic acid compound of formula 2b.

[0367] Solution 7

[0368]

[0369] In another synthetic variation, compounds of Formula 2a and Formula 2b can be prepared as described in Scheme 8. Morpholine of Formula G12 is directly converted to compound of Formula 2a by reductive amination or alkylation with the corresponding aldehyde of Formula G22 or electrophilic reagent of Formula G5, respectively, under the conditions described in Scheme 4. In the compound of Scheme 8 (where A is a protected carboxylic acid group, where PG 2 represents a suitable acid protecting group such as a methyl ester), subsequent deprotection of the ester of formula 2a can provide a compound of formula 2b.

[0370] Solution 8

[0371]

[0372] Intermediate aldehydes of formula G22 and alkylating agents of formula G5 can be prepared as described in Scheme 9. As described in Scheme 2, coupling of a boronic acid or boronic ester of formula G17 (wherein R is an alkyl group, such as methyl) with an aldehyde of formula G14 under Suzuki conditions provides an aldehyde intermediate of formula G22. Reduction of the aldehyde moiety of formula G22 by a reagent such as sodium borohydride in a solvent such as methanol provides an alcohol of formula G23. Alternatively, in a palladium-catalyzed Suzuki reaction as described above, an alcohol of formula G23 can be prepared by reaction of a boronic acid or boronic ester of formula G17 with an aromatic halide of formula G24 (wherein Hal is typically a bromide). The alcohol of formula G23 can be converted to a suitable alkylating agent of formula G5, wherein the leaving group LG is related to the nature of the subsequent alkylation reaction and is typically a bromide. Typical bromination conditions include treatment with 33% HBr in acetic acid at room temperature, heating with PBr in DCM at room temperature, and heating to room temperature. 3 or SOCl in DCM at room temperature 2 Those skilled in the art will appreciate that alternative leaving groups (including but not limited to chloride, mesylate and tosylate) may be formed and that alternative conditions will be required to synthesize such intermediates of formula G5.

[0373] Solution 9

[0374]

[0375] In other variations of compound 2a where U, V, W or Z is COH, compounds of formula G27 can be synthesized as described in Scheme 10. Phenol protected compounds of formula G25 (wherein PG is a suitable protecting group, typically methyl or benzyl) can be synthesized from morpholine of formula G12 as described in Scheme 4. Suzuki reaction between aryl halides of formula G25 (wherein Hal is typically bromide) and boronic acids or boronic esters of formula G17 under the conditions described in Scheme 4 affords bisaryl molecules of formula G26. Compounds (wherein A is a protected carboxylic acid group and PG is 2 Deprotection of the phenolic moiety and the protected acid moiety in (represents a suitable acid protecting group) produces a compound of formula G27. The protecting groups are usually methoxy and carbomethoxy, respectively, and typical methoxy deprotection conditions are boron tribromide in a solvent such as DCM at a temperature of -78°C to room temperature. Hydrolysis conditions using a hydroxide base are usually used to deprotect the ester moiety.

[0376] Solution 10

[0377]

[0378] In compound 2a (where R 5It's OSO 2 NH 2 In other variations (wherein U, V, W or Z is H or COH), compounds of formula 29 or formula 32 can be prepared as described in Scheme 11. As described herein, a phenol intermediate of formula G28 is first synthesized from a morpholine intermediate of formula G12. Sulfonamide acylation of the phenol group of formula G28 can be achieved under appropriate conditions, typically by treating with excess sulfamoyl chloride in DMA to obtain a final compound of formula G29. Alternatively, as described herein, a phenol-protected intermediate compound of formula G30 (wherein PG is a suitable protecting group, typically benzyl) can be synthesized from morpholine of formula G12. The sulfonamide acylation step as described above is performed to convert a compound of formula G28 into a compound of formula G29. The phenol protecting group of a compound of formula G31 is removed (typically by hydrogenating the benzyl group in a solvent such as ethanol in the presence of a catalyst such as palladium on carbon), and then a final compound of formula G32 is obtained.

[0379] Solution 11

[0380]

[0381] It should be understood that the above schemes and operations are not intended to be limited in any way. In fact, the above schemes and operations can also be used to prepare compounds of the present invention (wherein, for example, A is a carboxylic acid isostere group). The methods and protecting groups applicable to "A group" are well known to those skilled in the art, and for example, trityl can be used to protect tetrazole groups. In addition, a compound of formula 1 can be converted into another compound of the present invention by methods well known to technicians. Some examples of synthetic procedures for converting one functional group into another are described in standard texts such as March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 7th Edition, Michael B. Smith, John Wiley, 2013, (ISBN: 978-0-470-46259-1), Organic Syntheses, Online Edition, www.orgsyn.org, (ISSN 2333-3553) and Fiesers' Reagents for Organic Synthesis, Volumes 1 to 17, John Wiley, edited by Mary Fieser (ISBN: 0-471-58283-2).

[0382] In many of the above reactions, it may be necessary to protect one or more groups to prevent the reaction from occurring at undesirable locations on the molecule. Some examples of protecting groups and some methods of protecting and deprotecting functional groups can be found in Greene's Protective Groups in Organic Synthesis, Fifth Edition, Editor: Peter GMWuts, John Wiley, 2014, (ISBN: 9781118057483).

[0383] The compounds prepared by the aforementioned methods can be isolated and purified by any of a variety of methods known to those skilled in the art, and some examples of such methods include recrystallization and chromatographic techniques, such as column chromatography under normal or reverse phase conditions (e.g. flash chromatography), HPLC and SFC.

[0384] Example

[0385] The invention will now be illustrated, but not limited, by reference to the following examples.

[0386] General approach

[0387] In the absence of a preparation route, the relevant intermediates are commercially available. The data for the available intermediates are in Table 2. 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 to 27°C. 1H 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)-values ​​relative to the following solvents: chloroform-d=7.26ppm, DMSO-d6=2.50ppm, methanol-d4=3.31ppm. The following abbreviations are used for the multiplicity of NMR signals: s=singlet, br=broad, d=doublet, t=triplet, q=quadruplet, m=multiplet. Coupling constants are listed as J values, measured in Hz. NMR and mass spectrometry results were corrected to account for background peaks. Chromatography refers to the use of 60 to 120 mesh or 40 to 633 μm, Column chromatography was performed on silica gel and under nitrogen pressure (flash chromatography). Microwave mediated reactions were performed in Biotage initiators or CEM Discover microwave reactors.

[0388] LC / MS analysis

[0389] LC / MS analysis of the compounds was performed under electrospray conditions using the instrumentation and methods given below:

[0390] LC / MS Method A

[0391] Instrument: Acquity UPLC with photodiode array detector and QDA mass spectrometer detector; Column: Acquity C-18, 1.6 micron, 50×1.6 mm; Gradient [time (min) / solvent B in A (%)]: 0.00 / 10, 0.75 / 10, 2.80 / 90, 4.50 / 100, 4.60 / 100, 4.70 / 10, 6.00 / 10; 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 0.8 mL / min.

[0392] LC / MS Method B&C

[0393] Instrument: HP 1100 with G1315A DAD, Waters Micromass ZQ; Column: Phenomenex Gemini-NX C-18, 3 microns, 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 LH 2 O + 2.5 mL in H 2 28% ammonia in O solution; solvent B = 2.5LMeCN + 135mL H 2 O + 2.5 mL in H 2 28% ammonia in O 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.

[0394] LC / MS Method D

[0395] Instrument: HP 1100 with G1315ADAD, Waters Micromass ZQ; Column: Phenomenex Kinetex C-18, 5 micron, 2.1×50 mm; Gradient [time (min) / solvent B in A (%)]: 0.00 / 2, 0.01 / 2, 8.40 / 95, 10.00 / 95; Solvent: Solvent A = 0.1% formic acid in 10 mM ammonium formate aqueous solution; Solvent B = acetonitrile / water / formic acid (95:4.9:0.1). 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.

[0396] LC / MS Method E

[0397] 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 0.8 mL / min.

[0398] LC / MS Method F&G

[0399] Instrument: Acquity H-Class with photodiode array detector and QDA mass detector; Column: Acquity C-18, 1.6 micron, 50×1.6 mm; Method F gradient [time (min) / solvent B in A (%)]: 0.00 / 5, 0.20 / 5, 1.80 / 98, 2.00 / 100, 2.50 / 100, 2.51 / 5, 3.00 / 5; Method G 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.

[0400] LC / MS Method H

[0401] 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.

[0402] LC / MS Method I

[0403] Instrument: Shimadzu 2020 series with diode array detector; Column: Agilent Poroshell 120EC C-18, 2.7 micron, 4.6×50 mm; Gradient [time (min) / solvent B in A (%)]: 0.00 / 15, 1.00 / 15, 4.00 / 100, 4.50 / 100, 4.51 / 15, 5.00 / 15; 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.

[0404] HPLC purification

[0405] Where indicated, intermediates or final compounds were purified by reverse phase preparative HPLC conditions using the instrumentation and methods given below:

[0406] HPLC purification method A

[0407] Gilson Semi-Preparative HPLC System - includes 321 pump, 171 diode array detector and GX-271 liquid handler and Gilson Trilution software.

[0408] Preparative HPLC: [reversed phase (Phenomenex Kinetix C18, 100×30 mm, 5 μm, 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 containing 0.1% TFA. Solvent B: MeCN].

[0409] HPLC purification method B

[0410] Gilson Semi-Preparative HPLC System - includes 321 pump, 171 diode array detector and GX-271 liquid handler and Gilson Trilution software.

[0411] Preparative HPLC: [reversed phase (Gemini-NX C18, 100×30 mm, 5 μm, 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: 0.2% of 28% ammonia in water. Solvent B: MeCN].

[0412] HPLC Purification Method C

[0413] Gilson Semi-Preparative HPLC System - includes dual piston pumps 331 and 332, 171 diode array detector and GX-271 liquid handler and Gilson Trilution software.

[0414] Preparative HPLC: [reversed phase (Gemini-NX C18, 100×30 mm, 5 μm, 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: 0.2% of 28% ammonia in water. Solvent B: MeCN].

[0415] HPLC purification method D

[0416] Gilson Semi-Preparative HPLC System - includes dual piston pumps 331 and 332, 171 diode array detector and GX-271 liquid handler and Gilson Trilution software.

[0417] Preparative HPLC: [reversed phase (Phenomenex Kinetix C18, 100×30 mm, 5 μm, 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 containing 0.1% TFA. Solvent B: MeCN].

[0418] HPLC purification method E

[0419] Shimadzu preparative HPLC with LC-20Ap pump, SPD-20A diode array detector and CBM-20A controller.

[0420] Preparative HPLC: [reverse phase (Agilent Zorbax, 10 mL / min, gradient of solvent B in solvent A: 0% to 23% solvent B in 14 min, 98% solvent B for 2 min; solvent A: water with 0.1% TFA. Solvent B: MeCN].

[0421] HPLC purification method F

[0422] Shimadzu preparative HPLC with LC-20Ap pump, SPD-20A diode array detector and Labsolutions software.

[0423] Preparative HPLC: [reverse phase (Agilent 10C18, 250 x 21 mm, 20 mL / min, gradient of solvent B in solvent A: 5% to 80% solvent B; solvent A: water containing 0.1% TFA. Solvent B: MeCN].

[0424] HPLC purification method G

[0425] Shimadzu preparative HPLC with LC-20Ap pump, SPD-20A diode array detector and CBM-20A controller.

[0426] Preparative HPLC: [reverse phase (Agilent, 250×9.4 mm, 55 μm, 9 mL / min, gradient of solvent B in solvent A: 28% to 45% solvent B (in 18 min), 45% to 98% solvent B (for 2 min), 28% solvent B (for 2 min); solvent A: 5 mM aqueous ammonium bicarbonate solution. Solvent B: MeCN].

[0427] Synthesis of intermediates

[0428] Route 1, as exemplified by the preparation of Intermediate 1: methyl 4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate hydrochloride (Intermediate 1)

[0429]

[0430] Step (i): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (5.10 g, 22.1 mmol) and (S)-4-(1-aminoethyl)benzoic acid methyl ester hydrochloride (3.95 g, 22.1 mmol) in MeCN (70 mL) was added HATU (12.6 g, 33.1 mmol) and the mixture was stirred at room temperature for 30 minutes. After this, the mixture was cooled to 0°C and N,N-diisopropylethylamine (11.9 mL, 66.2 mmol) was added. The mixture was warmed to room temperature and stirred for 24 hours before being partitioned between EtOAc and water. The organic layer was separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4 The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (0% to 72%) in hexanes to afford (R)-tert-butyl 3-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (5.50 g, 14.0 mmol, 63%) as a sticky solid. (LC / MS Method A): m / z 293 [M+H-Boc] + (ES + ), at 2.17 minutes, UV active.

[0431] Step (ii): To a solution of (R)-tert-butyl 3-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (6.30 g, 16.1 mmol) in 1,4-dioxane (30 mL) was added 4N HCl in 1,4-dioxane (40 mL) at room temperature under nitrogen atmosphere. The mixture was stirred for 3 hours after which the solvent was removed under reduced pressure. The crude material was purified by elution from Et 2 O to give the intermediate methyl 1,4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate hydrochloride (5.30 g, 16.1 mmol, 100%) as an off-white solid. (LC / MS Method A): m / z 293 [M+H-HCl] + (ES + ), at 2.17 minutes, UV active.

[0432] Route 2, as exemplified by the preparation of Intermediate 2: 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 2)

[0433]

[0434] Step (i): 4-bromo-3-methylbenzoic acid (30 g, 0.14 mol) was dissolved in SOCl 2 The solution in (250mL) was stirred at 70°C for 15 hours. The mixture was cooled to room temperature and concentrated under vacuum. The residue was dissolved in DCM (50mL), cooled to 0°C and aqueous ammonium hydroxide solution (40mL) was added dropwise. The mixture was stirred at room temperature for 30 minutes, after which the solid was filtered and recrystallized from EtOH (250mL) to obtain 4-bromo-3-methylbenzamide (21.0g, 0.10mol, 70% yield) as a yellow solid. (LC / MS method I): m / z 214 [M+H] + (ES + ), at 2.76 minutes, UV active.

[0435] Step (ii): Under nitrogen atmosphere, 4-bromo-3-methylbenzamide (7.20 g, 33.6 mmol), bis(pinacolato)diboron (10.3 g, 40.4 mmol), potassium acetate (6.60 g, 67.3 mmol) and Pd(dppf)Cl 2 The mixture of (700mg, 0.96mmol) in 1,4-dioxane (150mL) was heated to 105°C for 15 hours. The mixture was cooled to room temperature and the solvent was removed under vacuum. The crude material was purified by flash column chromatography (normal phase, silica) in EtOAc (10% to 30%) gradient in petroleum ether to obtain intermediate 2,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8.40g, 32.2mmol, 96% yield) as a yellow solid. (LC / MS method I): m / z 262 [M+H] + (ES + ), at 4.84 minutes, UV active.

[0436] Route 3, as exemplified by the preparation of Intermediate 3: methyl 4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (Intermediate 3)

[0437]

[0438] Step (i): To a solution of (R)-morpholine-3-carboxylic acid hydrochloride (1.02 g, 6.13 mmol) and (S)-4-(1-aminoethyl)benzoate (1.0 g, 5.58 mmol) in MeCN (5 mL) was added HATU (3.18 g, 8.37 mmol) and the mixture was stirred at room temperature for 15 minutes. The mixture was cooled to 0°C and N,N-diisopropylethylamine (3.0 mL, 16.7 mmol) was added. The mixture was stirred at room temperature for 6 hours before being partitioned between EtOAc and water. The organics were separated and the aqueous layer was further extracted with EtOAc (×1). The combined organics were purified by Na 2 SO 4 The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (0% to 52%) in hexanes to afford the intermediate methyl 3,4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (0.80 g, 2.74 mmol, 49% yield) as a viscous liquid. (LC / MS Method A): m / z 293 [M+H] + (ES + ), at 1.33 minutes, UV active.

[0439] Route 4, as exemplified by the preparation of Intermediate 4: 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (Intermediate 4)

[0440]

[0441] Step (i): To a solution of 4-bromo-3-methylbenzenesulfonyl chloride (10.0 g, 37.1 mmol) in THF (75 mL) was added aqueous ammonia (75 mL) at 0°C. The mixture was stirred at the same temperature for 5 hours before being quenched by the addition of 1 N HCl. The mixture was extracted with DCM (×2) and the combined organics were concentrated to give 4-bromo-3-methylbenzenesulfonamide (9.15 g, 36.6 mmol, 99% yield) as a white powder. (LC / MS Method I): m / z 250 [M+H] + (ES + ), at 3.27 minutes, UV active.

[0442] Step (ii): Under nitrogen atmosphere, 4-bromo-3-methylbenzenesulfonamide (9.29 g, 37.2 mmol), bis(pinacolato)diboron (11.3 g, 44.6 mmol), potassium acetate (5.47 g, 55.8 mmol) and Pd(dppf)Cl 2The mixture of (900mg, 1.23mmol) in 1,4-dioxane (100mL) was heated to 105°C for 15 hours. The mixture was cooled to room temperature and the solvent was removed under vacuum. The crude material was purified by flash column chromatography (normal phase, silica) in EtOAc (20% to 25%) gradient in petroleum ether to obtain intermediate 4,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (6.32g, 21.3mmol, 57% yield) as a white solid. (LC / MS method I): m / z 298[M+H] + (ES + ), at 3.64 minutes, UV active.

[0443] Route 5, as exemplified by the preparation of Intermediate 5: 5-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylic acid methyl ester hydrochloride (Intermediate 5)

[0444]

[0445] Step (i): to the mixture of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (310mg, 1.34mmol) in DMF (5.5mL), N,N-diisopropylethylamine (1.16mL, 6.7mmol), HATU (613mg, 1.61mmol) and 5-(aminomethyl)-2-picolinic acid methyl ester dihydrochloride (405mg, 1.61mmol) are added. The mixture is stirred at room temperature for 18 hours, then diluted with EtOAc and water. The organic matter is separated, washed with salt water (×2), dried and concentrated via passing through a hydrophobic glass frit. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of MeOH (0% to 5%) in DCM to afford (R)-tert-butyl 3-(((6-(methoxycarbonyl)pyridin-3-yl)methyl)carbamoyl)morpholine-4-carboxylate (537 mg, 1.42 mmol, quantitative yield) as a light orange solid. (LC / MS Method B): m / z 380 [M+H] + (ES + ), at 1.67 minutes, UV active.

[0446] Step (ii): To a solution of (R)-tert-butyl 3-(((6-(methoxycarbonyl)pyridin-3-yl)methyl)carbamoyl)morpholine-4-carboxylate (510 mg, 1.34 mmol) in 1,4-dioxane (5 mL) was added 4M HCl in dioxane (5 mL, 1.34 mmol). The mixture was stirred at room temperature for 2 hours after which it was concentrated to give the intermediate methyl 5,5-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylate hydrochloride (425 mg, 1.34 mmol, 100% yield) as a white solid. (LC / MS Method B): m / z 280 [M+H-HCl] + (ES + ), at 0.37 min, UV active.

[0447] Route 6, as exemplified by the preparation of Intermediate 6 and Intermediate 9: 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 6) and 3'-formyl-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 9)

[0448]

[0449] Step (i): 3-Bromo-5-hydroxybenzaldehyde (4.00 g, 20.0 mmol), intermediate 2,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (5.74 g, 22.0 mmol) and potassium carbonate (3.20 g, 60.0 mmol) were dissolved in 1,4-dioxane (15 mL) and water (15 mL), and the mixture was purged with nitrogen for 30 minutes at room temperature. After this, PdCl was added. 2 (dppf). DCM (1.63 g, 2.00 mmol) and the reaction mixture was heated to 80 °C for 2 hours. The mixture was cooled and partitioned between EtOAc and water. The organics were separated and the aqueous layer was further extracted with EtOAc. The combined organics were washed with Na 2 SO 4 Dried and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0 to 35%) in water to give intermediate 9, 3'-formyl-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (3.60 g, 14.1 mmol, 71% yield) as a brown solid. (LC / MS Method F): m / z 256 [M+H] + (ES + ), at 1.09 minutes, UV active.

[0450] Step (ii): To a solution of 3'-formyl-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.50 g, 5.88 mmol) in methanol (15 mL) was added sodium borohydride (440 mg, 11.8 mmol) at 0°C. The reaction mixture was stirred at room temperature for 2 hours, after which it was dissolved in EtOAc and saturated NaHCO 3 The organics were separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4 Drying and concentration gave 3'-hydroxy-5'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.36 g, 5.29 mmol, 90% yield) as an off-white solid. (LC / MS Method F): m / z 258 [M+H] + (ES + ) at 0.94 minutes, UV active.

[0451] 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 acetic acid (12 mL) at room temperature. The mixture was stirred at room temperature for 16 hours, after which it was heated in saturated NaHCO 3 The aqueous solution was partitioned between EtOAc. The organics were separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were washed with Na 2 SO 4 Drying and concentration gave intermediate 6, 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.59 g, 4.97 mmol, 94% yield) as a reddish brown solid. (LC / MS Method F): m / z 321 [M+H] + (ES + ) at 1.23 minutes, UV active.

[0452] Route 7, as exemplified by the preparation of Intermediate 7: 3-Benzyloxy-5-bromo-benzaldehyde (Intermediate 7)

[0453]

[0454] Step (i): To a suspension of 3-bromo-5-hydroxybenzaldehyde (500 mg, 2.49 mmol) and potassium carbonate (1.03 g, 7.46 mmol) in DMF (6.5 mL) was added benzyl bromide (0.38 mL, 3.23 mmol). The mixture was stirred at room temperature for 18 hours before being partitioned between EtOAc and water. The organics were separated, washed sequentially with water and brine, dried through a hydrophobic frit and concentrated. The residue was purified by flash column chromatography (normal phase, silica) on EtOAc in isohexane. 2 Purification was performed with a gradient of 0 (0% to 10%) to afford intermediate 7, 3-benzyloxy-5-bromo-benzaldehyde (736 mg, 2.53 mmol, quantitative yield) as a viscous red oil. (LC / MS Method B): m / z 292 [M+H] + (ES + ) at 2.43 minutes, UV active.

[0455] Route 8, as exemplified by the preparation of Intermediate 8: methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (Intermediate 8)

[0456]

[0457] Step (i): (S)-4-(1-aminoethyl)-2-methoxybenzoic acid (2.00 g, 10.3 mmol) was dissolved in MeOH (20 mL) at room temperature, 4N HCl in 1,4-dioxane (10 mL) was added, and the reaction mixture was heated to reflux for 8 hours. The mixture was cooled, concentrated, and the obtained residue was ground from ether to obtain (S)-4-(1-aminoethyl)-2-methoxybenzoic acid methyl ester hydrochloride (2.10 g, 8.6 mmol, 83% yield) as a white solid.

[0458] 1 H NMR (400MHz, DMSO) δ1.53 (d, J=6.7Hz, 3H), 3.78 (s, 3H), 3.85 (s, 3H), 4.37-4.46 (m, 1H), 7 .13 (dd, J=7.9, 1.6Hz, 1H), 7.48 (d, J=1.6Hz, 1H), 7.66 (d, J=7.9Hz, 1H), 8.75 (br.s, 3H).

[0459] Step (ii): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (3.60 g, 15.5 mmol) and (S)-4-(1-aminoethyl)-2-methoxybenzoic acid methyl ester hydrochloride (3.25 g, 15.5 mmol) in DMF (35 mL) was added HATU (8.87 g, 23.3 mmol) and the mixture was stirred at room temperature for 15 minutes. The mixture was cooled to 0°C and N,N-diisopropylethylamine (8.12 mL, 46.6 mmol) was added. The mixture was stirred at room temperature for 4 hours after which it was partitioned between EtOAc and water. The organics were separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4 The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (0% to 48%) in hexanes to afford (R)-tert-butyl 3-(((S)-1-(3-methoxy-4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (6.20 g, 14.7 mmol, 95% yield) as a sticky solid. (LC / MS Method E): m / z 445 [M+Na] + (ES + ), at 1.78 minutes, UV active.

[0460] Step (iii): To a solution of (R)-tert-butyl 3-(((S)-1-(3-methoxy-4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (6.20 g, 14.7 mmol) in 1,4-dioxane (60 mL) was added 4 M HCl in 1,4-dioxane (60 mL) at room temperature and the mixture was stirred at room temperature for 2 hours. After this time, the mixture was stirred in EtOAc and saturated NaHCO 3 The organics were separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4 Drying and concentration gave methyl 2-methoxy-4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (3.20 g, 9.93 mmol, 68% yield) as a white solid. (LC / MS Method E): m / z 323 [M+H] + (ES + ), at 0.99 min, UV active.

[0461] Step (iv): To a solution of methyl 2-methoxy-4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (3.20 g, 9.93 mmol) in DCM at -78 °C was added BBr dropwise 3 (1M in DCM, 32 mL) and the mixture was stirred at the same temperature for 1 hour. After that, the mixture was stirred in EtOAc and saturated NaHCO 3 The organics were separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) in a gradient of MeCN (0% to 33%) in water to afford intermediate 8, methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (2.30 g, 7.46 mmol, 75% yield) as a white solid. (LC / MS Method E): m / z 309 [M+H] + (ES + ), at 1.34 minutes, UV active.

[0462] Route 9, as exemplified by the preparation of Intermediate 11: 3'-(bromomethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 11)

[0463]

[0464] Step (i): To a solution of intermediate 2,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (800 mg, 3.06 mmol) in 1,4-dioxane (26 mL) and water (6 mL) was added (3-bromophenyl)methanol (632 mg, 3.37 mmol), potassium acetate (900 mg, 9.18 mmol) and Pd(dppf)Cl under nitrogen atmosphere. 2 (224 mg, 0.306 mmol). The mixture was heated to 85 °C for 2.5 hours before it was concentrated under reduced pressure. The residue was partitioned between EtOAc and water, the organics were separated, and purified by Na 2 SO 4 Dried and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (5% to 23%) in DCM to give 3'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (400 mg, 1.65 mmol, 54% yield) as a brown solid. (LC / MS Method I): m / z 242 [M+H] +(ES + ), at 2.80 minutes, UV active.

[0465] Step (ii): To a solution of 3'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (300 mg, 1.24 mmol) in DCM (30 mL) was added PBr at 0 °C. 3 (503 mg, 1.86 mmol), and the mixture was stirred at room temperature for 2 hours. After this time, the mixture was poured into ice-cold water and extracted with DCM (×1). The organic matter was washed with Na 2 SO 4 Drying and concentration gave intermediate 11, 3'-(bromomethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (300 mg, 0.99 mmol, 80% yield) as a yellow solid. (LC / MS Method I): m / z 305 [M+H] + (ES + ), at 3.60 minutes, UV active.

[0466] Route 10, as exemplified by the preparation of Intermediate 12 and Intermediate 30: 4-(1-aminoethyl)-2,6-difluorobenzoic acid methyl ester hydrochloride (Intermediate 12 and Intermediate 30, isolated enantiomers, unidentified)

[0467]

[0468] Step (i): To a solution of 2,6-difluoro-4-formylbenzonitrile (5.00 g, 29.9 mmol) and (R)-tert-butylsulfonamide (3.63 g, 29.9 mmol) in anhydrous THF (60 mL) was added Ti(OEt) dropwise 4 (12.5 mL, 59.8 mmol). The mixture was stirred at room temperature for 16 hours, after which it was poured into brine. The resulting suspension was filtered through celite and washed with EtOAc. The filtrate was washed with brine and the organics were separated. The aqueous layer was further extracted with EtOAc (×2) and the combined organics were washed with Na 2 SO 4 Dry and concentrate. The residue was recrystallized from EtOAc / hexanes to give (R)-N-(4-cyano-3,5-difluorobenzylidene)-2-methylpropane-2-sulfenamide (5.00 g, 18.5 mmol, 62% yield) as a white solid. (LC / MS Method I): m / z 271 [M+H] + (ES + ), at 3.93 minutes, UV active.

[0469] Step (ii): To a solution of (R)-N-(4-cyano-3,5-difluorobenzylidene)-2-methylpropane-2-sulfenamide (5.50 g, 20.4 mmol) in anhydrous DCM (100 mL) was added methylmagnesium bromide (3 M in Et 2 O, 13.6 mL). The mixture was stirred at the same temperature for 1 hour, then warmed to room temperature and stirred at room temperature for 1 hour. The mixture was cooled to 0°C and stirred by slowly adding saturated NH 4 The organics were separated and the aqueous layer was extracted with EtOAc (×3). The combined organics were washed with Na 2 SO 4 Dry, concentrate, and recrystallize the residue from EtOAc / hexanes to give (R)-N-(1-(4-cyano-3,5-difluorophenyl)ethyl)-2-methylpropane-2-sulfenamide, a mixture of diastereomers (5.00 g, 17.5 mmol, 86% yield) as a yellow solid. (LC / MS Method I): m / z 287 [M+H] + (ES + ), at 3.48 minutes, UV active.

[0470] Step (iii): A suspension of (R)-N-(1-(4-cyano-3,5-difluorophenyl)ethyl)-2-methylpropane-2-sulfinamide, a mixture of diastereomers (350 mg, 1.22 mmol) and NaOH (147 mg, 3.67 mmol) was heated to 80°C for 5 hours before being cooled to room temperature, diluted with saturated aqueous citric acid and extracted with EtOAc (×3). The combined organics were washed with brine and purified by Na 2 SO 4 Drying and concentration gave 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoic acid, a mixture of diastereomers (350 mg, 1.15 mmol, 94% yield) as a yellow oil which was used without purification. (LC / MS Method I): m / z 306 [M+H] + (ES + ), at 2.60 minutes, UV active.

[0471] Step (iv): To a solution of 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoic acid, a mixture of diastereomers (2.66 g, 8.71 mmol) in MeOH (50 mL) at 0°C was added N'-ethylcarbodiimide hydrochloride (2.50 g, 13.1 mmol), 1-hydroxybenzotriazole hydrate (1.76 g, 13.1 mmol) and triethylamine (3.52 g, 34.8 mmol). The mixture was stirred at room temperature for 15 hours after which it was diluted with water and extracted with DCM (×3). The combined organics were washed with brine and purified by Na 2 SO 4 Dried and concentrated.The crude material is purified by flash column chromatography (normal phase, silica) under EtOAc (10% to 30%) gradient in petroleum ether to obtain fast eluting spot, as 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoic acid methyl esters, diastereomer 1 (1.00g, 3.13mmol, 36% yield) of yellow oil, and slow eluting spot, as 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoic acid methyl esters, diastereomer 2 (665mg, 2.08mmol, 24% yield) of yellow oil.

[0472] Diastereomer 1: (LC / MS method I): m / z 320 [M+H] + (ES + ), at 3.33 minutes, UV active.

[0473] Diastereomer 2: (LC / MS method I): m / z 320 [M+H] + (ES + ), at 3.24 minutes, UV active.

[0474] Step (v): To a solution of two separate methyl 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoate, each as a purified diastereomer (60 mg, 0.19 mmol) in MeOH (3 mL) was added 4 M HCl in 1,4-dioxane, and the mixture was stirred at room temperature for 1 hour. The solution was concentrated to give intermediate 12 as a yellow solid (47 mg, 0.19 mmol, 98% yield; (LC / MS Method I): m / z 216 [M+H-HCl] + (ES +), UV active at 1.66 min, or intermediate 30 as a yellow solid (45 mg, 0.18 mmol, 94% yield; (LC / MS Method I): m / z 315 [M+H-HCl] + (ES + ), at 1.49 minutes, UV active.

[0475] Route 11, as exemplified by the preparation of Intermediate 13: 4-bromo-3-((tert-butyldimethylsilyl)oxy)benzenesulfonamide (Intermediate 13)

[0476]

[0477] Step (i): To a solution of 4-bromo-3-methoxybenzenesulfonamide (1.00 g, 3.75 mmol) in DCM (10 mL) was added BBr at 0 °C. 3 (1M in DCM, 7.10 mL, 7.10 mmol). The mixture was stirred at room temperature for 16 h, after which it was dissolved in EtOAc and saturated NaHCO 3 The organics were separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4 Dry, concentrate under reduced pressure and the residue is purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 26%) in water to give 4-bromo-3-hydroxybenzenesulfonamide (0.70 g, 2.94 mmol, 74% yield) as an off-white solid. (LC / MS Method E): m / s 252 [M+H]H (ES + ), at 1.43 minutes, UV active.

[0478] Step (ii): A solution of 4-bromo-3-hydroxybenzenesulfonamide (100 mg, 0.39 mmol) and imidazole (81 mg, 1.19 mmol) in THF (2 mL) was stirred at -10 °C for 15 minutes, followed by the addition of tert-butyldimethylsilyl chloride (180 mg, 1.19 mmol) at the same temperature. The mixture was warmed to room temperature and stirred for 1 hour, after which it was partitioned between EtOAc and water. The organics were separated, and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4 Drying and concentration gave intermediate 13, 4-bromo-3-((tert-butyldimethylsilyl)oxy)benzenesulfonamide (160 mg, 0.43 mmol, quantitative yield) as an off-white solid which was used without purification.

[0479] 1H NMR (400MHz, DMSO) δ0.27 (s, 6H), 1.02 (s, 9H), 7.30-7.35 (m, 2H), 7.50 (br.s, 2H), 7.78-7.83 (m, 1H).

[0480] Route 12, as exemplified by the preparation of Intermediate 14: 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (Intermediate 14)

[0481]

[0482] Step (i): To a suspension of intermediate 2,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (4.00 g, 15.3 mmol), (5-bromopyridin-3-yl)methanol (2.88 g, 15.3 mmol) and potassium acetate (4.50 g, 45.9 mmol) in 1,4-dioxane (64 mL) and water (16 mL) was added Pd(dppf)Cl under nitrogen atmosphere. 2 To the 4- (5- (hydroxymethyl) pyridin-3-yl) -3-methylbenzamide (2.50 g, 10.3 mmol, 68% yield) of 4- (5- (hydroxymethyl) pyridin-3-yl) -3-methylbenzamide (1.12 g, 1.53 mmol) and the mixture was heated to 85 ° C for 16 hours. The mixture was cooled, filtered and concentrated under reduced pressure. The crude residue was purified by flash column chromatography (normal phase, silica) in MeOH (5% to 10%) gradient in DCM to give 4- (5- (hydroxymethyl) pyridin-3-yl) -3-methylbenzamide (2.50 g, 10.3 mmol, 68% yield) as a yellow solid. (LC / MS Method I): m / z 243 [M+H] + (ES + ), at 0.80 min, UV active.

[0483] Step (ii): To a solution of 4-(5-(hydroxymethyl)pyridin-3-yl)-3-methylbenzamide (1.20 g, 4.9 mmol) in DCM (30 mL) was added SOCl 2 To the mixture was added 4-(5-(chloromethyl)pyridin-3-yl)-3-methylbenzamide (1.0 g, 3.85 mmol, 78% yield) (LC / MS Method I): m / z 261 [M+H] + (ES + ), at 2.73 minutes, UV active.

[0484] Step (iii): To a suspension of 4-(5-(chloromethyl)pyridin-3-yl)-3-methylbenzamide (500 mg, 1.92 mmol) in DCM (15 mL) was added meta-chloroperbenzoic acid (664 mg, 3.84 mmol). The mixture was stirred at room temperature for 1 hour before being concentrated under reduced pressure. The crude residue was purified by flash column chromatography (normal phase, silica) in DCM with a gradient of MeOH (5% to 7%) to afford intermediate 14, 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (500 mg, 1.81 mmol, 94% yield) as a light yellow solid. (LC / MS Method I): m / z 277 [M+H] + (ES + ), at 2.30 minutes, UV active.

[0485] Route 13, as exemplified by the preparation of Intermediate 18: 6-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-3-carboxylic acid ethyl ester hydrochloride (Intermediate 18)

[0486]

[0487] Step (i): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (220 mg, 0.95 mmol) in DMF (4.5 mL) was added N,N-diisopropylethylamine (0.66 mL, 3.81 mmol), HATU (435.23 mg, 1.14 mmol) and 6-(aminomethyl)nicotinate ethyl ester hydrochloride (247 mg, 1.14 mmol). The mixture was stirred at room temperature for 18 hours before being partitioned between EtOAc and water. The organics were separated, washed with brine, dried and concentrated via passing through a hydrophobic frit. The crude residue was purified by flash column chromatography (normal phase, silica) with a gradient of MeOH (0% to 5%) in DCM to give (R)-tert-butyl 3-(((5-(ethoxycarbonyl)pyridin-2-yl)methyl)carbamoyl)morpholine-4-carboxylate (379 mg, 0.964 mmol, quantitative yield) as a light orange gum. (LC / MS Method B): m / z 394 [M+H] + (ES + ), at 1.90 min, UV active.

[0488] Step (ii): To a solution of tert-butyl (3R)-3-[(5-ethoxycarbonyl-2-pyridinyl)methylcarbamoyl]morpholine-4-carboxylate (379 mg, 0.96 mmol) in 1,4-dioxane (4 mL) was added 4M HCl in 1,4-dioxane (4 mL, 1.34 mmol). The mixture was stirred at room temperature for 2.5 hours after which it was concentrated to give intermediate 18, ethyl 6-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-3-carboxylate hydrochloride (330 mg, 1.00 mmol, quantitative yield) as a light brown solid. (LC / MS Method B): m / z 294 [M+H-HCl] + (ES + ), at 1.38 minutes, UV active.

[0489] Route 14, as exemplified by the preparation of Intermediate 20: methyl 2,6-difluoro-4-[[[(3R)-morpholine-3-carbonyl]amino]methyl]benzoate hydrochloride (Intermediate 20)

[0490]

[0491] Step (i): To a mixture of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (0.96 g, 4.13 mmol) in DMF (15 mL) was added N,N-diisopropylethylamine (2.38 mL, 13.8 mmol), HATU (1.84 g, 4.82 mmol) and (4-bromo-3,5-difluoro-phenyl)methylamine hydrochloride (890 mg, 3.44 mmol). The mixture was stirred at room temperature for 18 hours before being partitioned between EtOAc and water. The organics were separated, washed with brine, dried and concentrated via passing through a hydrophobic frit. The crude residue was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (20% to 80%) in isohexane to give (R)-tert-butyl 3-((4-bromo-3,5-difluorobenzyl)carbamoyl)morpholine-4-carboxylate (1.26 g, 2.89 mmol, 84% yield) as an off-white solid. (LC / MS Method B): m / z 436 [M+H] + (ES + ), at 2.18 minutes, UV active.

[0492] Step (ii): A mixture of (R)-tert-butyl 3-((4-bromo-3,5-difluorobenzyl)carbamoyl)morpholine-4-carboxylate (1.24 g, 2.85 mmol), triethylamine (0.48 mL, 3.42 mmol), palladium(II) acetate (64 mg, 0.29 mmol) and 1,1'-bis(diphenylphosphino)ferrocene (316 mg, 0.57 mmol) in MeOH (21 mL) and DMSO (6 mL) was purged with CO gas and then stirred at 80°C under an atmosphere of CO for 20 hours. The mixture was filtered through celite and concentrated. The residue was partitioned between EtOAc and water, and the organics were separated, washed with brine, purified by MgSO 4 The crude residue was purified by flash column chromatography (normal phase, silica) in a gradient of MeOH (0% to 10%) in DCM and then further purified by preparative HPLC [Purification Method B; 40% to 70% solvent B in solvent A] to give (R)-tert-butyl 3-((3,5-difluoro-4-(methoxycarbonyl)benzyl)carbamoyl)morpholine-4-carboxylate (302 mg, 0.73 mmol, 26% yield) as a light brown solid. (LC / MS Method B): m / z 415 [M+H] + (ES + ), at 2.02 minutes, UV active.

[0493] Step (iii): To a stirred solution of (R)-tert-butyl 3-((3,5-difluoro-4-(methoxycarbonyl)benzyl)carbamoyl)morpholine-4-carboxylate (302 mg, 0.73 mmol) in 1,4-dioxane (3 mL) was added 4M HCl in 1,4-dioxane (3 mL). The mixture was stirred at room temperature for 2.5 hours after which it was concentrated to give intermediate 20, methyl 2,6-difluoro-4-[[[(3R)-morpholine-3-carbonyl]amino]methyl]benzoate hydrochloride (247 mg, 0.71 mmol, 98% yield) as an off-white solid. (LC / MS Method B): m / z 294 [M+H-HCl] + (ES + ), at 1.38 minutes, UV active.

[0494] Route 15, as exemplified by the preparation of Intermediate 24: (4-carbamoyl-2-hydroxyphenyl)boronic acid (Intermediate 24)

[0495]

[0496] Step (i): 4-Bromo-3-hydroxybenzamide (1.00 g, 4.65 mmol), bis(pinacolato)diboron (2.36 g, 9.30 mmol) and potassium acetate (1.37 g, 14.0 mmol) were dissolved in 1,4-dioxane (10 mL) and the mixture was purged with nitrogen for 30 minutes. Pd(dppf)Cl was added 2 (43 mg, 0.05 mmol), and the reaction mixture was heated to 80 ° C for 4 hours. The mixture was concentrated under reduced pressure, and the residue was purified by flash column chromatography (reverse phase, C18) in water with a gradient of MeCN (0% to 2%) to give intermediate 24, (4-carbamoyl-2-hydroxyphenyl)boronic acid (0.51 g, 2.83 mmol, 61% yield) as an off-white solid. (LC / MS Method E): m / z 181 [M+H] + (ES + ), at 0.75 min, UV active.

[0497] Route 16, as exemplified by the preparation of Intermediate 32 and Intermediate 33: (R)-methyl 4-(1-(morpholine-3-amido)cyclopropyl)benzoate hydrochloride (Intermediate 32) and (R)-methyl 4-(1-(4-(3-bromo-4-hydroxybenzyl)morpholine-3-amido)cyclopropyl)benzoate (Intermediate 33)

[0498]

[0499] Step (i): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (3.50 g, 15.1 mmol) and methyl 4-(1-aminocyclopropyl)benzoate hydrochloride (4.13 g, 18.2 mmol) in DMF (35 mL) was added HATU (8.64 g, 22.7 mmol), and the mixture was stirred at room temperature for 15 minutes. The mixture was then cooled to 0°C, and N,N-diisopropylethylamine (7.91 mL, 45.4 mmol) was added, and then stirred at room temperature for 6 hours. The mixture was partitioned between EtOAc and water, and the organics were separated. The aqueous layer was further extracted with EtOAc (×2), and the combined organics were purified by Na 2 SO 4 The crude residue was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (0% to 40%) in hexanes to afford (R)-tert-butyl 3-((1-(4-(methoxycarbonyl)phenyl)cyclopropyl)carbamoyl)morpholine-4-carboxylate (5.50 g, 13.6 mmol, 90% yield) as a sticky solid. (LC / MS Method E): m / z 305 [M+H-Boc] +(ES + ), at 2.22 minutes, UV active.

[0500] Step (ii): Under nitrogen atmosphere, tert-butyl (R)-3-((1-(4-(methoxycarbonyl)phenyl)cyclopropyl)carbamoyl)morpholine-4-carboxylate (5.50 g, 13.6 mmol) was dissolved in 1,4-dioxane (55 mL) and 4N HCl in 1,4-dioxane (55 mL) was added at room temperature. The mixture was stirred at room temperature for 8 hours after which it was concentrated under reduced pressure. The crude material was purified by elution from Et 2 O to give Intermediate 32, (R)-methyl 4-(1-(morpholine-3-amido)cyclopropyl)benzoate hydrochloride (4.40 g, 12.9 mmol, 95% yield) as a white solid. (LC / MS Method E): m / z 305 [M+H-HCl] + (ES + ), at 1.07 minutes, UV active.

[0501] Step (iii): To a solution of intermediate 32, methyl (R)-4-(1-(morpholine-3-amido)cyclopropyl)benzoate hydrochloride (1.36 g, 4.00 mmol) and 3-bromo-4-hydroxybenzaldehyde (0.80 g, 4.00 mmol) in MeOH (8 mL) was added ZnCl at room temperature. 2 (0.27 g, 2.00 mmol), and the mixture was stirred at 70 ° C for 16 hours. The mixture was cooled to 0 ° C, sodium cyanoborohydride (0.75 g, 12.0 mmol) was added in portions, and the mixture was stirred at room temperature for 12 hours. After this time, the reaction was stirred in EtOAc and saturated NaHCO 3 The aqueous layer was further extracted with EtOAc (×2) and the combined organics were purified by Na 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) under a gradient of MeCN (0% to 65%) in water to afford intermediate 33, (R)-methyl 4-(1-(4-(3-bromo-4-hydroxybenzyl)morpholine-3-amido)cyclopropyl)benzoate (0.35 g, 0.72 mmol, 18% yield) as a brown solid. (LC / MS Method E): m / z 409 [M+H] + (ES + ), at 1.46 minutes, UV active.

[0502] Route 17, as exemplified by the preparation of Intermediate 35: (R)-N-((S)-1-(4-cyanophenyl)ethyl)morpholine-3-carboxamide hydrochloride (Intermediate 35)

[0503]

[0504] Step (i): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (0.50 g, 2.16 mmol) in acetonitrile (7 mL) was added (S)-4-(1-aminoethyl)benzonitrile hydrochloride (0.39 g, 2.16 mmol) followed by HATU (1.23 g, 3.24 mmol) and the reaction mixture was stirred at room temperature for 30 minutes. The mixture was then cooled to 0°C and N,N-diisopropylethylamine (1.13 mL, 6.49 mmol) was added and warmed to room temperature under stirring for 4 hours. The reaction mixture was partitioned between water (30 mL) and EtOAc (30 mL). The aqueous layer was further extracted with EtOAc (3×30 mL) and the combined organic layers were purified by Na 2 O 4 3-naphthalene pyridine (HPLC elution). 2 SO 4 The crude product was purified by gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 70% acetonitrile in water to give (R)-tert-butyl 3-(((S)-1-(4-cyanophenyl)ethyl)carbamoyl)morpholine-4-carboxylate (0.16 g, 21%) as a brown solid. (LC / MS Method E): m / z 260 [M+H-Boc] + (ES + ), at 2.29 minutes, UV active.

[0505] Step-(ii): To a solution of (R)-tert-butyl 3-(((S)-1-(4-cyanophenyl)ethyl)carbamoyl)morpholine-4-carboxylate (0.14 g, 0.40 mmol) in DCM (3 mL) was added 4N HCl in dioxane (2 mL) at 0°C and the reaction mixture was warmed and stirred at room temperature for 2 hours. The mixture was concentrated in vacuo to obtain the crude product which was triturated with diethyl ether (3 x 5 mL) to afford Intermediate 35 (R)-N-((S)-1-(4-cyanophenyl)ethyl)morpholine-3-carboxamide hydrochloride (0.113 g, 98%) as a grey-brown solid. (LC / MS Method A): m / z 260 [M+H-HCl] + (ES + ), at 0.81 min, UV active.

[0506] Intermediate 10: 4-((S)-1-((R)-4-((4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Intermediate 10)

[0507] Following the procedure described in Example 5, but using Intermediate 3 and (3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (Intermediate 25), the title compound was obtained. (LC / MS Method E): m / z 475 [M+H] + (ES + ), at 1.45 minutes, UV active.

[0508] Synthetic procedures of compounds of formula I

[0509] Example 1: Synthesis of 4-((S)-1-((R)-4-((5-(4-carbamoyl-2-methylphenyl)pyridin-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 1)

[0510]

[0511] Step (i): To a solution of intermediate 1,4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoic acid methyl ester hydrochloride (250 mg, 0.76 mmol) in DCM (3.5 mL) was added 5-bromonicotinaldehyde (283 mg, 1.52 mmol). The mixture was stirred at room temperature for 1 hour before sodium triacetoxyborohydride (338 mg, 1.6 mmol) was added. The mixture was stirred at room temperature for 17 hours before it was partitioned between DCM and 1 M NaOH (aq). The organics were separated and the aqueous layer was extracted with DCM (3x). The combined organics were washed with water then brine, dried via passing through a hydrophobic frit and concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (30% to 100%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[(5-bromo-3-pyridinyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (302 mg, 0.65 mmol, 86% yield) as a white solid. (LC / MS Method B): m / z 463 [M+H] + (ES + ), at 1.97 minutes, UV active.

[0512] Step (ii): A mixture of methyl 4-[(1S)-1-[[(3R)-4-[(5-bromo-3-pyridyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (150 mg, 0.32 mmol), intermediate 2,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (102 mg, 0.39 mmol), potassium carbonate (90 mg, 0.65 mmol) and tetrakis(triphenylphosphine)palladium(0) (37.5 mg, 0.03 mmol) in 1,4-dioxane (1.3 mL) and water (0.3 mL) was heated to 100° C. under microwave irradiation for 1 hour. The mixture was diluted with water and EtOAc, and the organics were separated, washed with water, then brine, dried via passing through a hydrophobic frit, and concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of MeOH (0% to 10%) in DCM to afford methyl 4-[(1S)-1-[[(3R)-4-[[5-(4-carbamoyl-2-methyl-phenyl)-3-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (143.2 mg, 0.27 mmol, 85% yield) as a light yellow foam. (LC / MS Method B): m / z 517 [M+H] + (ES + ), at 1.81 minutes, UV active.

[0513] Step (iii): A mixture of methyl 4-[(1S)-1-[[(3R)-4-[[5-(4-carbamoyl-2-methyl-phenyl)-3-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (143 mg, 0.28 mmol) and lithium hydroxide monohydrate (40.7 mg, 0.97 mmol) in 1,4-dioxane (2 mL) and water (2 mL) was stirred at room temperature for 3 hours. The mixture was concentrated to dryness and the crude material was purified by preparative HPLC [Purification Method A] to give 4-[(1S)-1-[[(3R)-4-[[5-(4-carbamoyl-2-methyl-phenyl)-3-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (28.5 mg, 0.06 mmol, 20% yield) as a white solid.

[0514] 1H NMR (DMSO, 400MHz): δ (ppm) 1.34 (d, J=7.0Hz, 3H), 2.14-2.22 (m, 1H), 2.23 (s, 3H), 2.66-2.72 (m, 1 H), 3.02 (dd, J=9.1, 3.5Hz, 1H), 3.34 (d, J=13.7Hz, 1H), 3.45-3.56 (m, 2H), 3.64-3.70 (m, 1H), 3.7 5 (dd, J=11.1, 3.5Hz, 1H), 3.79 (d, J=13.7Hz, 1H), 4.91-5.00 (m, 1H), 7.25-7.31 (m, 3H), 7.34 (brs , 1H), 7.75-7.80(m, 4H), 7.82-7.84(m, 1H), 8.07(brs, 1H), 8.44-8.48(m, 2H), 8.50-8.53(m, 1H).

[0515] No COO observed H Proton. (LC / MS method D): m / z 503 [M+H] + (ES + ) at 1.67 minutes, UV active.

[0516] Example 5: Synthesis of 4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 5)

[0517]

[0518] Step (i): At room temperature, the intermediate 3,4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoic acid methyl ester (390 mg, 1.33 mmol), potassium carbonate (740 mg, 5.33 mmol) and sodium iodide (20 mg, 0.13 mmol) were dissolved in MeCN (5 mL). 1-Bromo-3-(bromomethyl)benzene (280 mg, 1.12 mmol) was added and the reaction mixture was stirred at 80 ° C for 1 hour. The mixture was cooled and distributed between EtOAc and water. The organics were separated and the aqueous layer was further extracted with EtOAc. The combined organics were purified by Na 2 SO 4The residue was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (0% to 68%) in hexanes to afford methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-amido)ethyl)benzoate (400 mg, 0.87 mmol, 78% yield) as an off-white solid. (LC / MS Method A): m / z 461 [M+H] + (ES + ) at 2.16 minutes, UV active.

[0519] Step (ii): Methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-amido)ethyl)benzoate (0.20 g, 0.43 mmol), intermediate 4,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (0.15 g, 0.52 mmol) and potassium carbonate (0.12 g, 0.86 mmol) were dissolved in a mixture of 1,4-dioxane (2 mL) and water (2 mL), and the mixture was purged with nitrogen for 30 minutes at room temperature. PdCl2(dppf).DCM (0.07 g, 0.09 mmol) was added, and the reaction mixture was heated to 80°C for 3 hours. After this, the mixture was cooled and partitioned between EtOAc and water. The organics were separated, and the aqueous layer was further extracted with EtOAc. The combined organics were purified by Na 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 48%) in water to give methyl 4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.16 g, 0.29 mmol, 67% yield) as an off-white solid. (LC / MS Method A): m / z 552 [M+H] + (ES + ) at 1.90 min, UV active.

[0520] Step (iii): Methyl 4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.16 g, 0.29 mmol) was dissolved in 1,4-dioxane (3 mL) and water (1 mL). Lithium hydroxide monohydrate (60 mg, 1.45 mmol) was added at room temperature and the reaction mixture was stirred for 3 hours before being partitioned between EtOAc and water. The organic layer was separated and the aqueous layer was acidified to pH 1 using 4N HCl(aq). The acidic aqueous layer was extracted with EtOAc (×2) and these combined organics were purified by Na 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) under a gradient of MeCN (0% to 38%) in water to give 4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (68 mg, 0.13 mmol, 44% yield) as a white solid.

[0521] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.37 (d, J=7.0Hz, 3H), 2.09-2.20 (m, 1H), 2.25-2.35 (m, 4H), 2.66-2.73(m, 1H), 2.96-3.06(m, 1H), 3.21-3.29(m, 1H), 3.44-3.59(m, 2H), 3. 65-3.83(m, 3H), 4.93-5.06(m, 1H), 7.24-7.28(m, 1H), 7.35-7.44(m, 7H), 7.65-7.7 3(m, 1H), 7.73-7.78(m, 1H), 7.84-7.93(m, 2H), 8.50-8.60(m, 1H), 12.84(br.s, 1H).

[0522] (LC / MS method A): m / z 538 [M+H] + (ES + ), at 1.65 minutes, UV active.

[0523] Example 9: Synthesis of 5-[[[(3R)-4-[[3-(4-carbamoyl-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylic acid (Compound 9)

[0524]

[0525] Step (i): To a solution of intermediate 5,5-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylic acid methyl ester hydrochloride (218 mg, 0.69 mmol) in DCM (3.2 mL) was added 3-bromobenzaldehyde (0.16 mL, 1.38 mmol). The mixture was stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (307 mg, 1.45 mmol) was added. The mixture was stirred at room temperature for another 17 hours, after which it was concentrated in vacuo. The residue was suspended in saturated NaHCO3 (aqueous solution) and extracted with EtOAc (×3). The combined organic phases were washed with water and brine, dried through a hydrophobic glass frit, and concentrated in vacuo. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of MeOH (0% to 10%) in DCM to give methyl 5-[[[(3R)-4-[(3-bromophenyl)methyl]morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylate (216 mg, 0.48 mmol, 70% yield) as a colorless gum. (LC / MS Method B): m / z 449 [M+H] + (ES + ), at 1.92 minutes, UV active.

[0526] Step (ii): A mixture of 5-[[[(3R)-4-[(3-bromophenyl)methyl]morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylic acid methyl ester (216 mg, 0.48 mmol), intermediate 2,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (163 mg, 0.63 mmol), potassium carbonate (200 mg, 1.44 mmol) and tetrakis(triphenylphosphine)palladium(0) (56 mg, 0.05 mmol) in 1,4-dioxane (1.6 mL) and water (0.4 mL) was heated to 100 ° C in a microwave reactor for 4 hours. The mixture was diluted with water and EtOAc, and the organics were separated. The aqueous layer was adjusted to pH 5 using concentrated HCl and then extracted with EtOAc. Both the organic phase and the pH 5 aqueous phase were concentrated in vacuo, and the residues were combined and purified by preparative HPLC [Purification Method D] to give 5-[[[(3R)-4-[[3-(4-carbamoyl-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylic acid (107 mg, 45%) as a white foam.

[0527] 1 H NMR (CD 3OD, 400MHz): δ (ppm) 2.30 (s, 3H), 3.31-3.44 (m, 2H), 3.65-3.78 (m, 2H), 3.99-4.06 (m, 1H), 4.12-4.18 (m, 1H), 4.20-4.28 (m, 2 H), 4.53-4.61 (m, 3H), 7.30 (d, J=7.9Hz, 1H), 7.45-7.57 (m, 4H), 7.71-7.78 (m, 1H), 7.80-7.85 (m, 1H), 7.96-8.03 (m, 1H), 8.11 -8.19(m,1H),8.63-8.69(m,1H).

[0528] No exchangeable protons were observed. (LC / MS method C): m / z 489 [M+H] + (ES + ), at 1.88 minutes, UV active.

[0529] Example 10: 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 10)

[0530]

[0531] Step (i): To a solution of intermediate 1,4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate hydrochloride (1.14 g, 3.47 mmol) and N,N-diisopropylethylamine (1.81 mL, 10.4 mmol) in MeCN (11 mL) was added intermediate 6,3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.11 g, 3.47 mmol) at room temperature, and the mixture was heated to 70°C for 5 hours. After cooling to room temperature, the mixture was partitioned between water and EtOAc. The aqueous layer was separated and extracted with EtOAc (×2). The combined organic layers were purified by Na 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 52%) in water to give methyl 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (1.62 g, 3.05 mmol, 88% yield) as a brown solid. (LC / MS Method F): m / z 532 [M+H] + (ES +), at 1.11 minutes, UV active.

[0532] Step (ii): Methyl 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (1.60 g, 3.04 mmol) was dissolved in a mixture of 1,4-dioxane (6 mL) and water (4 mL). Lithium hydroxide monohydrate (0.64 g, 15.2 mmol) was added at room temperature and the reaction mixture was stirred for 16 hours. The mixture was then partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH 2 with 1N HCl aqueous solution and then extracted with EtOAc (×3). The combined organic layers were purified by NaOH extraction. 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) under a gradient of MeCN (0% to 43%) in water to give 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (1.06 g, 2.05 mmol, 68% yield) as an off-white solid.

[0533] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.37 (d, J=7.0Hz, 3H), 2.09-2.18 (m, 1H), 2.25 (s, 3H), 2.69-2.78 (m, 1H), 2. 97-3.03 (m, 1H), 3.13 (d, J=13.1Hz, 1H), 3.43-3.59 (m, 2H), 3.65-3.82 (m, 3H), 4.88-5.06 (m, 1H), 6.60 (s , 1H), 6.72-6.80 (m, 2H), 7.23 (d, J=7.9Hz, 1H), 7.34 (s, 1H), 7.42 (d, J=7.9Hz, 2H), 7.72 (d, J=7.9Hz, 1H ), 7.79 (s, 1H), 7.86 (d, J=7.9Hz, 2H), 7.95 (s, 1H), 8.49 (d, J=7.8Hz, 1H), 9.51 (s, 1H), 12.83 (br.s, 1H).

[0534] (LC / MS method G): m / z 518 [M+H] + (ES + ), at 1.60 min, UV active.

[0535] Example 11: 4-[(1S)-1-[[(3R)-4-[[3-hydroxy-5-(2-methyl-4-sulfamoyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 11)

[0536]

[0537] Step (i): To a solution of intermediate 1,4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoic acid methyl ester hydrochloride (1.01 g, 3.07 mmol) in MeCN (35 mL) was added 1-bromo-3-(bromomethyl)-5-methoxybenzene (1.72 g, 6.14 mmol) and potassium carbonate (1.06 g, 7.68 mmol). The resulting mixture was heated to 70 ° C for 18 hours, then cooled to room temperature, filtered through a phase separator and concentrated. The residue was dissolved in EtOAc, washed with water and brine, dried and concentrated by passing through a hydrophobic frit. The residue was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (30% to 90%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[(3-bromo-5-methoxy-phenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (1.35 g, 2.74 mmol, 89% yield) as a white solid. (LC / MS Method B): m / z 492 [M+H] + (ES + ), at 2.32 minutes, UV active.

[0538] Step (ii): A mixture of intermediate 4,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (330 mg, 1.11 mmol), methyl 4-[(1S)-1-[[(3R)-4-[(3-bromo-5-methoxy-phenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (455 mg, 0.93 mmol), potassium carbonate (256 mg, 1.85 mmol) and tetrakis(triphenylphosphine)palladium(0) (107 mg, 0.09 mmol) in 1,4-dioxane (6 mL) and water (4 mL) was heated to 100° C. in a microwave reactor for 30 minutes. The mixture was diluted with water and EtOAc and the organics were separated, washed with water and brine, dried via passing through a hydrophobic frit and concentrated. The residue was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (30% to 100%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[[3-methoxy-5-(2-methyl-4-sulfamoyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (500 mg, 0.86 mmol, 93% yield) as an off-white solid. (LC / MS Method B): m / z 582 [M+H] + (ES + ), at 2.10 minutes, UV active.

[0539] Step (iii): To a solution of methyl 4-[(1S)-1-[[(3R)-4-[[3-methoxy-5-(2-methyl-4-sulfamoyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (505 mg, 0.87 mmol) in DCM (12 mL) was added a 1 M solution of boron tribromide in DCM (6.08 mL, 6.08 mmol) dropwise under a nitrogen atmosphere at -78°C. The mixture was gradually warmed to room temperature and stirred at room temperature for 18 hours. The mixture was cooled to 0°C and MeOH / H2O was added dropwise. 2 The mixture was stirred at room temperature for 1 hour, after which it was concentrated. The crude residue was purified by preparative HPLC [purification method D] to give 4-[(1S)-1-[[(3R)-4-[[3-hydroxy-5-(2-methyl-4-sulfamoyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (68.1 mg, 0.12 mmol, 14% yield) as an off-white foam.

[0540] 1 H NMR (CD 3OD, 400MHz): δ (ppm) 1.51 (d, J = 7.1Hz, 3H), 2.27 (s, 3H), 3.23-3.28 (m, 1H), 3.34-3.35 ( m, 1H), 3.36-3.42 (m, 1H), 3.70-3.78 (m, 2H), 3.99-4.10 (m, 2H), 4.20-4.29 (m, 2H), 5.1 1(d, J=7.1Hz, 1H), 6.73-6.75(m, 1H), 6.83-6.85(m, 1H), 6.87-6.89(m, 1H), 7.32(d, J= 8.0Hz, 1H), 7.44-7.52(m, 2H), 7.73-7.77(m, 1H), 7.81-7.82(m, 1H), 7.94-7.98(m, 2H).

[0541] No exchangeable protons were observed. (LC / MS method D): m / z 554 [M+H] + (ES + ), at 1.63 minutes, UV active.

[0542] Example 15: 4-[(1S)-1-[[(3R)-4-[[3-hydroxy-5-(2-methyl-4-sulfamoyloxy-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 15)

[0543]

[0544] Step (i): To a solution of intermediate 1,4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoic acid methyl ester hydrochloride (300 mg, 0.91 mmol) in DCM (4.2 mL) was added intermediate 7,3-benzyloxy-5-bromo-benzaldehyde (531 mg, 1.82 mmol). The mixture was stirred at room temperature for 1 hour, after which sodium triacetoxyborohydride (406 mg, 1.92 mmol) was added. The mixture was stirred at room temperature for 19 hours, after which it was partitioned between 1M NaOH (aq.) and DCM. The organics were separated, and the aqueous layer was further extracted with DCM (3×). The organic phases were combined, washed with water and brine, dried and concentrated via passing through a hydrophobic frit. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (30% to 80%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[(3-benzyloxy-5-bromo-phenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (424 mg, 0.74 mmol, 82% yield) as a white solid. (LC / MS Method B): m / z 568 [M+H] + (ES+ ), at 2.59 minutes, UV active.

[0545] Step (ii): A mixture of methyl 4-[(1S)-1-[[(3R)-4-[(3-benzyloxy-5-bromo-phenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (450 mg, 0.79 mmol), (4-hydroxy-2-methylphenyl)boronic acid (145 mg, 0.95 mmol), potassium carbonate (219 mg, 1.59 mmol) and tetrakis(triphenylphosphine)palladium(0) (91.6 mg, 0.08 mmol) in 1,4-dioxane (3.2 mL) and water (0.8 mL) was heated to 100° C. in a microwave reactor for 2 hours. Additional tetrakis(triphenylphosphine)palladium(0) (45.8 mg, 0.04 mmol), potassium carbonate (55 mg, 0.4 mmol) and (4-hydroxy-2-methylphenyl)boric acid (72 mg, 0.48 mmol) were added and the mixture was heated to 100 °C for an additional 30 minutes. The mixture was partitioned between water and EtOAc and the organics were separated, washed with water and brine, dried via passing through a hydrophobic frit and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) in a gradient of EtOAc (30% to 90%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[[3-benzyloxy-5-(4-hydroxy-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (413 mg, 0.69 mmol, 88% yield) as an orange gum. (LC / MS method B): m / z 595 [M+H] + (ES + ), at 2.45 minutes, UV active.

[0546] Step (iii): To a solution of methyl 4-[(1S)-1-[[(3R)-4-[[3-benzyloxy-5-(4-hydroxy-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (413 mg, 0.69 mmol) in water (4 mL) and 1,4-dioxane (4 mL) was added lithium hydroxide monohydrate (102 mg, 2.43 mmol). The mixture was stirred at room temperature for 3 hours after which it was concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (30% to 90%) in isohexane containing 0.1% v / v acetic acid to give 4-[(1S)-1-[[(3R)-4-[[3-benzyloxy-5-(4-hydroxy-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (67 mg, 0.12 mmol, 17% yield) as an off-white foam. (LC / MS Method B): m / z 581 [M+H] + (ES + ), at 1.66 minutes, UV active.

[0547] Step (iv): To a solution of 4-[(1S)-1-[[(3R)-4-[[3-benzyloxy-5-(4-hydroxy-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (67 mg, 0.12 mmol) in DMA (1 mL) was added sulfamoyl chloride (33.4 mg, 0.29 mmol). The mixture was stirred at room temperature for 18 hours before additional sulfamoyl chloride (20 mg, 0.17 mmol) was added and the mixture was stirred for an additional 18 hours. Water (0.75 mL) was added and the mixture was stirred for 30 minutes before it was partitioned between EtOAc and 1 M HCl(aq). The organics were separated and the aqueous layer was further extracted with EtOAc (3x). The combined organics were concentrated and triturated from EtOAc to give 4-[(1S)-1-[[(3R)-4-[[3-benzyloxy-5-(2-methyl-4-sulfamoyloxy-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (30 mg, 0.04 mmol, 37% yield) as a colorless glass. (LC / MS Method B): m / z 660 [M+H] + (ES + ), at 1.45 minutes, UV active.

[0548] Step (v): To a solution of 4-[(1S)-1-[[(3R)-4-[[3-benzyloxy-5-(2-methyl-4-sulfamoyloxy-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (30 mg, 0.05 mmol) in ethanol (0.45 mL) was added 10% palladium on carbon (1 mg). The mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere, after which the suspension was passed through a plug of celite and washed with methanol. The filtrate was concentrated under reduced pressure and the crude residue was purified by preparative HPLC [Purification Method D] to give 4-[(1S)-1-[[(3R)-4-[[3-hydroxy-5-(2-methyl-4-sulfamoyloxy-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (18.5 mg, 0.033 mmol, 71% yield) as a colorless glass.

[0549] 1 H NMR (CD 3 OD, 400MHz): δ (ppm) 1.51 (d, J=7.0Hz, 3H), 2.21 (s, 3H), 3.24-3.29 (m, 1H), 3.36-3.44 (m, 1H), 3.69-3.79 (m, 2H), 3.99-4.10 (m, 3H), 4. 18-4.31 (m, 2H), 5.10 (q, J=7.0Hz, 1H), 6.69-6.71 (m, 1H), 6.81-6.85 (m, 2H), 7.18-7.25 (m, 3H), 7.45-7.49 (m, 2H), 7.95-7.99 (m, 2H).

[0550] No exchangeable protons were observed. (LC / MS method D): m / z 570 [M+H] + (ES + ), at 2.76 minutes, UV active.

[0551] Example 18: 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoic acid (Compound 18)

[0552]

[0553] Step (i): Under nitrogen atmosphere, intermediate 8, methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (0.20 g, 0.65 mmol) and intermediate 9, 3'-formyl-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (0.33 g, 1.30 mmol) were dissolved in methanol (2 mL). At room temperature, the mixture was added Molecular sieves (0.5 g) and glacial acetic acid (4 μL, 0.065 mmol) were added, and the mixture was stirred for 4 hours. After that, the reaction mixture was cooled to 0 ° C, and sodium cyanoborohydride (0.12 g, 1.95 mmol) was added in batches, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated, and the residue was heated in saturated NaHCO 3 The aqueous solution was partitioned between EtOAc. The aqueous layer was separated and further extracted with EtOAc (3×). The combined organics were purified by Na 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) in a gradient of MeCN (0 to 44%) in water to afford methyl 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoate (0.19 g, 0.34 mmol, 54% yield) as a white solid. (LC / MS Method E): m / z 548 [M+H] + (ES + ) at 1.37 minutes, UV active.

[0554] Step (ii): Methyl 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoate (0.19 g, 0.34 mmol) was dissolved in 1,4-dioxane (2 mL) and water (2 mL). Lithium hydroxide monohydrate (73 mg, 1.75 mmol) was added and the mixture was stirred at room temperature for 12 hours. The solvent was removed under reduced pressure and the crude material was dissolved in 4N aqueous HCl, adsorbed onto celite and purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 24%) in water to give 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoic acid (0.11 g, 0.21 mmol, 60% yield) as a white solid.

[0555] 1 H NMR (DMSO, 400MHz): 8 (ppm) 1.34 (d, J=7.0Hz, 3H), 2.11-2.31 (m, 4H), 2.68-2.79 (m, 1H), 2.99-3.0 9(m, 1H), 3.11-3.20(m, 1H), 3.45-3.60(m, 2H), 3.65-3.85(m, 3H), 4.83-4.95(m, 1H), 6.61(s, 1H), 6.74-6.79(m, 2H), 6.82-6.90(m, 2H), 7.23(d, J=7.9Hz, 1H), 7.33(s, 1H), 7.66-7.74(m, 2H), 7.79( d, J=1.8Hz, 1H), 7.95 (s, 1H), 8.51 (br.s, 1H), 9.53 (br.s, 1H), 11.75 (br.s, 1H), 13.76 (br.s, 1H).

[0556] (LC / MS method E): m / z 534 [M+H] + (ES + ), at 1.22 minutes, UV active.

[0557] Example 19: 4-((S)-1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoic acid (Compound 19)

[0558]

[0559] Step (i): To a solution of intermediate 8, methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (500 mg, 1.62 mmol) and N,N-diisopropylethylamine (0.57 mL, 3.25 mmol) in MeCN (5 mL) was added 1-bromo-3-(bromomethyl)benzene (490 mg, 1.95 mmol) and the mixture was heated to 80 ° C for 2 hours. The mixture was cooled and partitioned between EtOAc and water. The organic layer was separated and the aqueous layer was further extracted with EtOAc (×2). The combined organics were purified by Na 2 SO 4The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (0% to 34%) in hexanes to afford methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-amido)ethyl)-2-hydroxybenzoate (550 mg, 1.15 mmol, 71% yield) as a white solid. (LC / MS Method E): m / z 478 [M+H] + (ES + ), at 1.94 minutes, UV active.

[0560] Step (ii): Methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-amido)ethyl)benzoate (250 mg, 0.53 mmol), intermediate 2,3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (692 mg, 2.65 mmol) and potassium carbonate (146 mg, 1.06 mmol) were dissolved in a mixture of 1,4-dioxane:water (1:1, 6 mL) and the mixture was purged with nitrogen at room temperature for 30 minutes. After this, PdCl was added. 2 (dppf). DCM (43 mg, 0.05 mmol) and the reaction mixture was heated to 80 °C for 2 hours. The mixture was cooled and partitioned between EtOAc and water. The organics were separated and the aqueous layer was further extracted with EtOAc. The combined organics were washed with Na 2 SO 4 The residue was purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 58%) in water to afford methyl 4-((S)-1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoate (170 mg, 0.32 mmol, 61%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), at 1.52 minutes, UV active.

[0561] Step (iii): To a solution of methyl 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amide)ethyl)-2-hydroxybenzoate (170 mg, 0.32 mmol) in 1,4-dioxane (2 mL) and water (2 mL) was added lithium hydroxide monohydrate (73 mg, 1.60 mmol), and the mixture was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure and the residue was acidified with 4N HCl, directly adsorbed onto celite and purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 33%) in water to give 4-((S)-1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoic acid (65 mg, 0.13 mmol, 39% yield) as an off-white solid.

[0562] 1 H NMR (400MHz, DMSO) δ1.34 (d, J=7.0Hz, 3H), 2.11-2.28 (m, 4H), 2.68-2.77 (m, 1H), 3.00- 3.08(m, 1H), 3.24-3.29(m, 1H), 3.47-3.58(m, 2H), 3.64-3.88(m, 3H), 4.85-4.96(m, 1H) , 6.80-6.95 (m, 2H), 7.18-7.30 (m, 2H), 7.30-7.48 (m, 4H), 7.69 (d, J=8.1Hz, 1H), 7.72-7 .76 (m, 1H), 7.81 (d, J=1.8Hz, 1H), 7.97 (s, 1H), 8.53 (d, J=8.0Hz, 1H), 12.47 (br.s, 2H).

[0563] (LC / MS method E): m / z 518 [M+H] + (ES + ), at 1.35 minutes, UV active.

[0564] Example 20: 4-((S)-1-((R)-4-((2'-methyl-4'-(sulfamoyloxy)-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 20)

[0565]

[0566] Step (i): To a solution of intermediate 10, 4-((S)-1-((R)-4-((4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (65 mg, 0.13 mmol) in DMA (2 mL) was added sulfamoyl chloride (157 mg, 1.37 mmol) at room temperature. The mixture was stirred at room temperature for 3 hours before being concentrated under reduced pressure. The residue was purified by flash column chromatography (reverse phase, C18) in a gradient of MeCN (0% to 48%) in water to give 4-((S)-1-((R)-4-((2'-methyl-4'-(sulfamoyloxy)-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (25 mg, 0.05 mmol, 33% yield) as an off-white solid.

[0567] 1 H NMR (400MHz, DMSO) δ1.37 (d, J=7.0Hz, 3H), 2.11-2.19 (m, 1H), 2.22 (s, 3H), 2.6 5-2.74 (m, 1H), 3.00 (dd, J=9.3, 3.5Hz, 1H), 3.23 (d, J=13.2Hz, 1H), 3.44-3.57 ( m, 2H), 3.65-3.81 (m, 3H), 4.92-5.04 (m, 1H), 7.12-7.33 (m, 6H), 7.35-7.47 (m, 3 H), 7.82-7.90 (m, 2H), 8.03 (br.s, 2H), 8.54 (d, J=8.1Hz, 1H), 12.98 (br.s, 1H).

[0568] (LC / MS method E): m / z 554 [M+H] + (ES + ), at 1.51 minutes, UV active.

[0569] Example 23: 4-[(1S)-1-[[(3R)-4-[[3-(4-carbamoylphenyl)-4-hydroxy-phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 23)

[0570]

[0571] Step (i): A mixture of intermediate 1,4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoic acid methyl ester hydrochloride (600 mg, 1.82 mmol) and 3-bromo-4-methoxybenzaldehyde (785 mg, 3.65 mmol) in DCM (8.5 mL) was stirred at room temperature for 1 hour before sodium triacetoxyborohydride (812 mg, 3.83 mmol) was added. The mixture was stirred at room temperature for 16 hours before it was washed with saturated NaHCO 3 The mixture was diluted with water. The organics were separated, washed with water and brine, dried over a hydrophobic frit and concentrated. The residue was purified by flash column chromatography (normal phase, silica) in a gradient of EtOAc (30% to 100%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[(3-bromo-4-methoxy-phenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (619 mg, 1.26 mmol, 69% yield) as a white solid. (LC / MS Method B): m / z 492 [M+H] + (ES + ), at 2.27 minutes, UV active.

[0572] Step (ii): A mixture of methyl 4-[(1S)-1-[[(3R)-4-[(3-bromo-4-methoxy-phenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (250 mg, 0.51 mmol), 4-aminocarbonylphenylboronic acid (101 mg, 0.61 mmol), potassium carbonate (141 mg, 1.02 mmol) and tetrakis(triphenylphosphine)palladium(0) (59 mg, 0.05 mmol) in 1,4-dioxane (2.4 mL) and water (0.6 mL) was heated to 100° C. in a microwave reactor for 40 minutes. Additional tetrakis(triphenylphosphine)palladium(0) (29 mg, 0.03 mmol), 4-aminocarbonylphenylboronic acid (25 mg, 0.15 mmol) and potassium carbonate (35 mg, 0.25 mmol) were added and the mixture was heated to 100° C. in a microwave reactor for an additional 70 minutes. The mixture was partitioned between water and EtOAc, and the organics were separated, washed with water and brine, dried via passing through a hydrophobic frit and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) under a gradient of MeOH (0% to 10%) in DCM to give methyl 4-[(1S)-1-[[(3R)-4-[[3-(4-carbamoylphenyl)-4-methoxy-phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (328 mg, 0.61 mmol, quantitative yield) as a light orange solid. (LC / MS Method B): m / z 532 [M+H] + (ES+ ), at 2.00 min, UV active.

[0573] Step (iii): To a solution of methyl 4-[(1S)-1-[[(3R)-4-[[3-(4-carbamoylphenyl)-4-methoxy-phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (270 mg, 0.51 mmol) in DCM (8 mL) was added a 1 M solution of boron tribromide in DCM (2.54 mL, 2.54 mmol) dropwise at -78 °C under a nitrogen atmosphere. The mixture was gradually warmed to room temperature and stirred at room temperature for 18 hours. The mixture was cooled to 0 °C and MeOH / H2O was added dropwise. 2 To the 4-[(1S)-1-[[(3R)-4-[[3-(4-carbamoylphenyl)-4-hydroxy-phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (54mg, 0.11mmol, 21% yield) of white foam was added.

[0574] 1 H NMR (CD 3 OD, 400MHz): δ (ppm) 1.51 (d, J = 7.0Hz, 3H), 3.22-3.28 (m, 1H), 3.35-3.43 (m, 1H), 3 .67-3.78(m, 2H), 3.98-4.05(m, 2H), 4.08(d, J=12.9Hz, 1H), 4.20-4.29(m, 2H), 5.0 8-5.16 (m, 1H), 6.93 (d, J=8.3Hz, 1H), 7.15 (dd, J=8.3, 2.2Hz, 1H), 7.30 (d, J=2.2H z, 1H), 7.47-7.53 (m, 2H), 7.62-7.67 (m, 2H), 7.87-7.92 (m, 2H), 7.98-8.02 (m, 2H).

[0575] No exchangeable protons were observed. (LC / MS method H): m / z 504 [M+H] + (ES + ), at 1.64 minutes, UV active.

[0576] Example 26: 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2,6-difluorobenzoic acid, single diastereomer of unknown stereochemistry (Compound 26)

[0577]

[0578] Step (i): (R)-morpholine-3-carboxylic acid methyl ester hydrochloride (220 mg, 1.21 mmol), intermediate 11, 3'-(bromomethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (360 mg, 1.19 mmol) and NaHCO 3 A mixture of (R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxylic acid methyl ester (420 mg, 1.14 mmol, 96% yield) was obtained as a yellow solid. (LC / MS Method I): m / z 369 [M+H] + (ES + ), at 3.00 min, UV active.

[0579] Step (ii): To a solution of (R)-methyl 4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxylate (420 mg, 1.14 mmol) in MeOH (9 mL) was added 1 M aqueous LiOH (3.42 mL, 3.42 mmol) and the mixture was heated to 50 °C for 3 hours. After this time, the mixture was concentrated, the residue was dissolved in water, and the pH was adjusted to 7 with 1 N HCl. The precipitate was filtered to give (R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxylic acid (320 mg, 0.90 mmol, 79% yield) as a white solid. (LC / MS Method I): m / z 355 [M+H] + (ES + ), at 1.97 minutes, UV active.

[0580] Step (iii): To a suspension of (R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxylic acid (150 mg, 0.42 mmol) in DCM at 0°C was added hydroxybenzotriazole (86 mg, 0.64 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (121 mg, 1.69 mmol). The mixture was stirred at room temperature for 1 hour before triethylamine (171 mg, 1.69 mmol) and intermediate 1,2,4-(1-aminoethyl)-2,6-difluorobenzoic acid methyl ester (107 mg, 0.42 mmol) were added at 0°C. The mixture was stirred at room temperature for 12 hours before it was diluted with DCM, washed with 0.5N HCl, and purified by Na 2 SO 4 The residue was purified by flash column chromatography (normal phase, silica), 3% MeOH in DCM to give methyl 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2,6-difluorobenzoate (130 mg, 0.24 mmol, 56% yield) as a white solid. (LC / MS Method I): m / z 552 [M+H] + (ES + ), at 3.10 minutes, UV active.

[0581] Step (iv): To a solution of methyl 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2,6-difluorobenzoate (130 mg, 0.24 mmol) in MeOH (10 mL) was added 1 M aqueous LiOH (0.71 mL, 0.71 mmol). The mixture was stirred at room temperature for 4 days before it was concentrated and partitioned between EtOAc and water. The aqueous layer was separated and adjusted to pH 4 with 1 N HCl before it was extracted with EtOAc (3×). The organics were combined and purified by Na 2 SO 4 The crude material was purified by preparative HPLC [Purification Method F] to give 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2,6-difluorobenzoic acid, single diastereomer of unknown stereochemistry 1 as a white solid (50.0 mg, 0.09 mmol, 40% yield).

[0582] 1 H NMR (CD3 OD, 400MHz): δ (ppm) 1.51 (d, J = 7.0Hz, 3H), 2.36 (s, 3H), 3.34-3.44 (m, 2H), 3.59-3.76 (m, 2H), 3.99-4.13 (m, 2H), 4.18-4.29 (m, 2H), 4.57- 4.72 (m, 1H), 5.07 (q, J=7.0, 1H), 6.98-7.08 (m, 2H), 7.37 (d, J=8.0, 1H), 7.52-7.66 (m, 4H), 7.79 (dd, J=8.0, 1.8, 1H), 7.86 (d, J=1.8, 1H).

[0583] No exchangeable protons were observed. (LC / MS method I): m / z 538 (M+H] + (ES + ), at 0.50 min, UV active.

[0584] Example 29: 4-((S)-1-((R)-4-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 29)

[0585]

[0586] Step (i): To a solution of intermediate methyl 3,4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate (300 mg, 1.03 mmol) and 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (390 mg, 1.54 mmol) in MeOH (3 mL) was added under nitrogen atmosphere. Molecular sieves and glacial acetic acid (6 μL, 0.1 mmol). The mixture was stirred at room temperature for 4 hours, after which it was cooled to 0°C and sodium cyanoborohydride (190 mg, 3.08 mmol) was added in portions. The mixture was stirred at room temperature for 16 hours, after which it was heated in saturated NaHCO 3 The aqueous solution was partitioned between EtOAc. The organics were separated and the aqueous layer was further extracted with EtOAc. The combined organics were purified by Na 2 SO 4The residue was purified by flash column chromatography (reverse phase, C18) under a gradient of MeCN (0% to 52%) in water to give methyl 4-((S)-1-((R)-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine-3-amido)ethyl)benzoate (108 mg, 0.21 mmol, 21% yield) as a white solid. (LC / MS Method E): m / z 427 [M+H-82] + (ES + )(mass of boric acid), at 1.50 min, UV active.

[0587] Step (ii): Methyl 4-((S)-1-((R)-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine-3-amido)ethyl)benzoate (90 mg, 0.17 mmol), intermediate 13,4-bromo-3-((tert-butyldimethylsilyl)oxy)benzenesulfonamide (78 mg, 0.21 mmol) and potassium carbonate (48 mg, 0.35 mmol) were dissolved in a mixture of 1,4-dioxane (3 mL) and water (1 mL). The mixture was purged with nitrogen at room temperature for 20 minutes, after which PdCl was added. 2 (dppf). DCM (14 mg, 0.026 mmol) and the reaction mixture was heated to 80 °C for 2 hours. The mixture was cooled and partitioned between water and EtOAc. The organics were separated and the aqueous layer was further extracted with EtOAc. The combined organics were washed with Na 2 SO 4 Dried and concentrated, and the residue was purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 53%) in water to give methyl 4-((S)-1-((R)-4-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (45 mg, 0.081 mmol, 46% yield) as a white solid. (LC / MS Method E): m / z 554 [M+H] + (ES + ), at 1.28 minutes, UV active.

[0588] Step (iii): To a solution of methyl 4-((S)-1-((R)-4-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (45 g, 0.08 mmol) in 1,4-dioxane (2 mL) and water (1 mL) was added lithium hydroxide monohydrate (17 mg, 0.40 mmol), and the mixture was stirred at room temperature for 3 hours. After this time, the mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (reverse phase, C18) with a gradient of MeCN (0% to 28%) in water to give 4-((S)-1-((R)-4-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (19 mg, 0.035 mmol, 44% yield) as a white solid.

[0589] 1 H NMR (400MHz, DMSO) δ1.39 (d, J=7.0Hz, 3H), 2.11-2.20 (m, 1H), 2.65-2.76 (m, 1H), 3.00 (dd, J=9.2, 3.6Hz, 1H), 3.22 (d, J=13.1Hz, 1H), 3.46-3.57 (m, 2H), 3.62-3.82(m, 3H), 4.90-5.04(m, 1H), 7.18-7.55(m, 10H), 7.51(s, 1H), 7.86 (d, J=8.0Hz, 2H), 8.51 (d, J=8.0Hz, 1H), 10.34 (br.s, 1H), 12.97 (br.s, 1H).

[0590] (LC / MS method E): m / z 540 [M+H] + (ES + ), at 1.44 minutes, UV active.

[0591] Example 30: 3-(4-Carbamoyl-2-methylphenyl)-5-(((R)-3-(((S)-1-(4-carboxyphenyl)ethyl)carbamoyl)morpholino)methyl)pyridine 1-oxide (Compound 30)

[0592]

[0593] Step (i): To a solution of intermediate 14, 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (200 mg, 0.72 mmol) in MeCN (12 mL) and DMF (2 mL) was added intermediate 1, methyl 4-((S)-1-((R)-morpholine-3-amido)ethyl)benzoate hydrochloride (238 mg, 0.72 mmol), NaHCO 3 To the 4- (4- (carbamoyl) -2- methylphenyl) -5- (((R) -3- (((S) -1- (4- (methoxycarbonyl) phenyl) ethyl) carbamoyl) morpholino) methyl) pyridine 1- oxide (326mg, 0.61mmol, 85% yield) as white solid. (LC / MS method I): m / z 533 [M+H] + (ES + ), at 2.73 minutes, UV active.

[0594] Step (ii): To a solution of 3-(4-carbamoyl-2-methylphenyl)-5-(((R)-3-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholino)methyl)pyridine 1-oxide (320 mg, 0.60 mmol) in MeOH (10 mL) was added 1 M aqueous lithium hydroxide solution (1.8 mL, 1.8 mmol) and the mixture was stirred at room temperature for 48 hours. After this time, the mixture was diluted with water and acidified to pH 5 with 1 N HCl. The solvent was removed under reduced pressure and the crude material was purified by preparative HPLC [Purification Method F] to give 3-(4-carbamoyl-2-methylphenyl)-5-(((R)-3-(((S)-1-(4-carboxyphenyl)ethyl)carbamoyl)morpholino)methyl)pyridine 1-oxide (90 mg, 0.17 mmol, 29% yield) as a white solid.

[0595] 1 H NMR (400 MHz, CD 3OD) δ1.51 (d, J=7.0Hz, 3H), 2.34 (s, 3H), 2.83-3.11 (m, 1H), 3.19-3.29 (m, 1H), 3.65-4.03 (m, 5H), 4.05-4.25 (m, 2H), 5.11 (q, J=7.0Hz , 1H), 7.29-7.38(m, 1H), 7.42-7.52(m, 2H), 7.61(s, 1H), 7.77-7.84(m, 1H), 7.88(s, 1H), 7.92-7.99(m, 2H), 8.40(s, 1H), 8.50(s, 1H).

[0596] No exchangeable protons were observed. (LC / MS method I): m / z 519 [M+H] + (ES + ) at 2.73 minutes, UV active.

[0597] Example 35: 4-((S)-1-((R)-4-((4'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 35)

[0598]

[0599] Step (i): To a solution of intermediate 1,4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoic acid methyl ester hydrochloride (500mg, 1.52mmol) in MeCN (12mL) was added potassium carbonate (525mg, 3.8mmol) and 3-bromobenzyl bromide (intermediate 28, 760mg, 3.0mmol). The reaction mixture was heated to 70°C for 18 hours, after which the mixture was cooled and filtered through a phase separator. The solution was concentrated and the residue was distributed between EtOAc and water. The organics were separated, washed with brine, dried and concentrated via a hydrophobic glass frit. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (30% to 70%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[(3-bromophenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (574 mg, 1.24 mmol, 82% yield) as a white solid. (LC / MS Method B): m / z 462 [M+H] + (ES + ), at 2.29 minutes, UV active.

[0600] Step (ii): A solution of 4-[(1S)-1-[[(3R)-4-[(3-bromophenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid methyl ester (150 mg, 0.33 mmol), tetrakis(triphenylphosphine)palladium(0) (37.6 mg, 0.03 mmol), potassium carbonate (89.9 mg, 0.65 mmol) and 4-hydroxyphenylboronic acid (53.8 mg, 0.39 mmol) in 1,4-dioxane (1.3 mL) and water (0.3 mL) was heated to 100 ° C in a microwave reactor for 1 hour. After this time, the mixture was diluted with EtOAc and water. The organics were separated, washed with brine, dried and concentrated via passing through a hydrophobic frit. The crude material was purified by flash column chromatography (normal phase, silica) with a gradient of EtOAc (30% to 100%) in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[[3-(4-hydroxyphenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (90 mg, 0.19 mmol, 58.3% yield) as a white solid. (LC / MS Method B): m / z 475 [M+H] + (ES + ), at 2.15 minutes, UV active.

[0601] Step (iii): A mixture of lithium hydroxide monohydrate (31.8 mg, 0.76 mmol) and methyl 4-[(1S)-1-[[(3R)-4-[[3-(4-hydroxyphenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (90 mg, 0.19 mmol) in THF (1.5 mL) and water (1.5 mL) was stirred at room temperature for 16 hours. The mixture was concentrated and the crude residue was purified by reverse phase preparative HPLC [Purification Method B] to give 4-((S)-1-((R)-4-((4'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (25 mg, 0.054 mmol, 29% yield) as a gum which was scratched to give an amorphous white solid.

[0602] 1 H NMR (CD 3OD, 400MHz): δ (ppm) 1.49 (d, J = 7.0Hz, 3H), 3.20-3.25 (m, 1H), 3.30-3.40 (m, 1H), 3.60- 3.75 (m, 2H), 3.90-4.06 (m, 2H), 4.12 (d, J=12.8Hz, 1H), 4.23 (dd, J=12.7, 3.9Hz, 1H), 4 .29(d, J=12.8Hz, 1H), 5.05-5.16(m, 1H), 6.77-6.89(m, 2H), 7.14-7.24(m, 1H), 7.36-7 .43(m, 3H), 7.45-7.49(m, 2H), 7.51-7.55(m, 1H), 7.60-7.65(m, 1H), 7.96-8.01(m, 2H).

[0603] No exchangeable protons were observed. (LC / MS method D): m / z 461 [M+H] + (ES + ), at 2.23 minutes, UV active.

[0604] Example 36: (R)-N-((S)-1-(4-(1H-tetrazolyl-5-yl)phenyl)ethyl)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamide (Compound 36)

[0605]

[0606] Step (i): To a solution of (R)-N-((S)-1-(4-cyanophenyl)ethyl)morpholine-3-carboxamide hydrochloride (Intermediate 35, 0.080 g, 0.27 mmol) and DIPEA (0.11 mL, 0.67 mmol) in acetonitrile (3 mL) was added 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 6, 0.086 g, 0.27 mmol) at room temperature and the reaction mixture was stirred at 70°C for 3 hours. The reaction mixture was then partitioned between water (20 mL) and EtOAc (20 mL). The aqueous layer was extracted with EtOAc (3 x 15 mL). The organic layers were combined and purified by Na 2 SO 4The 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 55% acetonitrile in water to give (R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-N-((S)-1-(4-cyanophenyl)ethyl)morpholine-3-carboxamide (0.085 g, 63%) as a brown solid. (LC / MS Method E): m / z 499 [M+H] + (ES + ), at 1.54 minutes, UV active.

[0607] Step (ii): To a solution of (R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)-N-((S)-1-(4-cyanophenyl)ethyl)morpholine-3-carboxamide (0.060 g, 0.12 mmol) in DMF (3 mL) was added sodium azide (0.156 g, 2.40 mmol) and ammonium chloride (0.128 g, 2.40 mmol) at room temperature and the reaction mixture was heated to 180°C for 16 hours. The reaction mixture was cooled to room temperature and then partitioned between water (20 mL) and dichloromethane (20 mL). The aqueous layer was further extracted with dichloromethane (3 x 20 mL). The organic layers were combined and dried (Na2O3). 2 SO 4 The solvent was removed in vacuo to give the crude product which was purified by reverse phase flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 60% acetonitrile in water to give (R)-N-((S)-1-(4-(4-(1H-tetrazol-5-yl)phenyl)ethyl)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamide (0.017 g, 26%) as a white solid.

[0608] 1H NMR (400Mz, DMSO) δ (ppm) 1HNMR: (400Mz, DMSO) 1.37 (d, 3H, J = 7.2Hz), 2.13 (s, 1H), 2.26 (s, 3H), 2.54-2.46 (m, 1H), 2.74-2.71 (m, 1H), 2.99 (q, 1H, J=8.8Hz), 3.14 (d, 1H, J=20.0Hz), 3.53-3.47 (m, 2H), 3.76-3.67 (m, 3H), 4.98-4.94 (m, 1H), 6.59 (s, 1H), 6.76 (d, 2H, J=7.2Hz), 7.22 (d, 1H, J=8.0Hz), 7.35 (d, 3H, J=10.0Hz), 7.70 (d, 1H, J=8.0Hz), 7.77 (s, 1H), 7.88 (d, 2H, J=8.0Hz), 8.01 (s, 1H), 8.38 (d, 1H, J=8.0Hz), 9.55 (s, 1H).

[0609] Tetrazolyl N was not observed H . (LC / MS method E): m / s 542 [M+H] + (ES + ), at 1.52 minutes, UV active.

[0610] Example 37: 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 37)

[0611]

[0612] Step (i): (S)-Morpholine-3-carboxylic acid hydrochloride (2.50 g, 14.97 mmol) was dissolved in acetone (15 mL) and H 2 O (15 mL). Add K 2 CO 3 The mixture was stirred at 0 ° C for 30 minutes. BOC anhydride (5.22 g, 23.95 mmol) was added and the reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was concentrated in vacuo and the subsequent residue was distributed between water (200 mL) and DCM (100 mL). The aqueous layer was further extracted with DCM (2 × 70 mL) and the combined organic layers were dried (Na 2 SO 4 ). 2 SO 4The solvent was removed in vacuo to give (S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (2.80 g, 88%) as a white solid. (LC / MS Method E): m / z 132 [M (-100) + H] + (ES + ), at 1.32 minutes, UV active.

[0613] Step (ii): (S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (2.8 g, 12.12 mmol) was dissolved in DMF (20 mL), and (S)-4-(1-aminoethyl)benzoic acid methyl ester (2.60 g, 14.53 mmol) was added to the reaction mixture at room temperature. HATU (6.90 g, 18.17 mmol) was then added, and the reaction mixture was stirred at room temperature for 30 minutes. After this, N,N-diisopropylethylamine (6.27 mL, 36.34 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then distributed between water (250 mL) and EtOAc (100 mL), and the aqueous layer was further extracted with EtOAc (2×70 mL). The organic layers were combined and dried (Na 2 SO 4 ). 2 SO 4 The crude material was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 70% acetonitrile in water to give (S)-tert-butyl 3-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (3.50 g, 79.4%) as a white solid. (LC / MS Method E): m / z 293 [M (-100) + H] + (ES + ), at 1.82 minutes, UV active.

[0614] Step (iii): Under nitrogen atmosphere, (S)-tert-butyl 3-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)-morpholine-4-carboxylate (3.50 g, 8.92 mmol) was dissolved in dioxane (10 mL). After this, 4N HCl in dioxane (10 mL) was added at room temperature and the reaction mixture was stirred at room temperature for 4 hours. The solvent was removed in vacuo and the crude material was purified by trituration with diethyl ether (15 mL) to give methyl 4-((S)-1-((S)-morpholine-3-amido)ethyl)benzoate hydrochloride (2.60 g, 89.0%) as a white solid. (LC / MS Method E): m / z 293 [M+H] + (ES + ), at 0.96 min, UV active.

[0615] Step (iv): Methyl 4-((S)-1-((S)-morpholine-3-amido)ethyl)benzoate hydrochloride (0.35 g, 1.06 mmol) and DIPEA (0.55 mL, 3.20 mmol) were dissolved in acetonitrile (7 mL) at room temperature. After this, 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 6, 0.40 g, 1.28 mmol) was added and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was concentrated 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 70% acetonitrile in water to give methyl 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.25 g, 44.6%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), at 1.31 minutes, UV active.

[0616] Step (v): Methyl 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.210 g, 0.395 mmol) was dissolved in dioxane (3 mL) and water (2 mL). Lithium hydroxide monohydrate (0.082 g, 1.98 mmol) was added at room temperature and stirred at room temperature for 4 hours. The reaction mixture was acidified with glacial acetic acid (1.5 mL) to adjust the pH to about 4 and concentrated in vacuo. The crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 35% acetonitrile in water to give 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (0.150 g, 73.4%) as a white solid.

[0617] 1H NMR: (400MHz, DMSO): δ (ppm) 1.36 (d, 3H, J = 6.8Hz), 2.15-2.11 (m, 1H), 2.25 (s, 3H), 2.74 (d, 1H, J = 12.0Hz), 2.99 (dd, 1H, J = 3.2Hz and J=9.2Hz), 3.14(d, 1H, J=13.6Hz), 3.53-3.47(m, 2H), 3.70-3.66(m, 2H) , 3.78-3.76 (m, 1H), 5.01 (t, 1H, J=7.4Hz), 6.61 (s, 1H), 6.76 (d, 2H, J=10 .4Hz), 7.23 (d, 1H, J=8Hz), 7.37 (t, 3H, J=8.4Hz), 7.73 (d, 1H, J=8.0Hz), 7.80 (d, 3H, J=8.4Hz), 7.96 (s, 1H), 8.50 (d, 1H, J=8.4Hz), 9.52 (s, 1H).

[0618] No carboxylic acid protons were observed. (LC / MS method E): m / z 518 [M+H] + (ES + ), at 1.14 minutes, UV active.

[0619] Example 38: 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 38)

[0620]

[0621] Step (i): (S)-Morpholine-3-carboxylic acid hydrochloride (2.50 g, 14.96 mmol) was dissolved in acetone (15 mL) and H 2 O (15mL). Potassium carbonate (10.40g, 74.83mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. After that, the reaction mixture was cooled to 0°C and BOC anhydride (5.22g, 23.95mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 5 hours and concentrated in vacuo. The residue was distributed between water (100mL) and DCM (50mL), and the aqueous layer was further extracted with DCM (2×50mL). The organic layers were combined and dried (Na 2 SO 4 ). The solvent was removed in vacuo to give (S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (2.80 g, 81.2%) as a white solid. (LC / MS Method E): m / z 132 [M (-100) + H]+ (ES + ), at 1.32 minutes, UV active.

[0622] Step (ii): (S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (2.10 g, 9.09 mmol) was dissolved in DMF (20 mL), and (R)-4-(1-aminoethyl)methyl benzoate (1.95 g, 10.90 mmol) and HATU (5.18 g, 13.64 mmol) were added, and the reaction mixture was stirred at room temperature for 30 minutes. After this, N,N-diisopropylethylamine (4.70 mL, 27.25 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was distributed between water (100 mL) and EtOAc (30 mL), and the aqueous layer was further extracted with EtOAc (2×50 mL). The organic layers were combined and dried (Na 2 SO 4 ). 2 SO 4 ) and the resulting crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 60% acetonitrile in water to afford (S)-tert-butyl 3-(((R)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (3.00 g, 84.3%) as a white solid. (LC / MS Method E): m / z 293 [M(-100)+H] + (ES + ), at 1.74 minutes, UV active.

[0623] Step (iii): Under nitrogen atmosphere, tert-butyl (S)-3-(((R)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (3.00 g, 7.64 mmol) was dissolved in dioxane (10 mL). After this, 4N HCl in dioxane (10 mL) was added at room temperature and the reaction mixture was stirred at room temperature for 3 hours. The solvent was removed in vacuo and the crude product was purified by trituration with diethyl ether (15 mL) to give methyl 4-((R)-1-((S)-morpholine-3-amido)ethyl)benzoate hydrochloride (2.50 g, 100%) as a white solid. (LC / MS Method E): m / z 293 [M+H] + (ES + ), at 0.95 min, UV active.

[0624] Step (iv): Methyl 4-((R)-1-((S)-morpholine-3-amido)ethyl)benzoate hydrochloride (0.35 g, 1.06 mmol) and DIPEA (0.55 mL, 3.20 mmol) were dissolved in acetonitrile (7 mL) at room temperature. After this, 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 6, 0.33 g, 1.28 mmol) was added and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was concentrated in vacuo and the crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product from 0% to 70% acetonitrile in water to afford methyl 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.35 g, 62.50%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), at 1.23 minutes, UV active.

[0625] Step (v): Methyl 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.35 g, 0.66 mmol) was dissolved in dioxane (1 mL) and water (2 mL). Lithium hydroxide monohydrate (0.13 g, 3.29 mmol) was added at room temperature and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was acidified with glacial acetic acid (2 mL) to adjust the pH to about 4 and concentrated in vacuo. The crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 35% acetonitrile in water to give 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (0.175 g, 51.5%) as a white solid.

[0626] 1H NMR: (400MHz, DMSO): δ (ppm) 1.37 (d, 3H, J = 6.8Hz), 2.16-2.08 (m, 1H), 2.25 (s, 3H), 2.73 (d, 1H, J = 11.6Hz), 2.99 (dd, 1H, J = 3.2Hz and J=9.2Hz), 3.13 (d, 1H, J=12.8Hz), 3.56-3.48 (m, 2H), 3.76-3.69 (m, 3H), 4.9 8 (t, 1H, J=7.4Hz), 6.60 (s, 1H), 6.74-6.77 (m, 2H), 7.23 (d, 1H, J=8.0Hz), 7. 34(s, 1H), 7.42(d, 2H, J=8.4Hz), 7.72(d, 1H, J=7.6Hz), 7.79(s, 1H), 7.86(d , 2H, J=8.4Hz), 7.96 (s, 1H), 8.49 (d, 1H, J=8Hz), 9.52 (s, 1H), 12.87 (s, 1H).

[0627] (LC / MS method E): m / z 518 [M+H] + (ES + ), at 1.00 min, UV active.

[0628] Example 39: 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 39)

[0629]

[0630] Step (i): (R)-Morpholine-3-carboxylic acid hydrochloride (10.00 g, 59.87 mmol) was dissolved in acetone (50 mL) and H 2 O (50mL). Potassium carbonate (41.37g, 299.31mmol) was added, and the reaction mixture was stirred at 0°C for 30 minutes. After this, BOC anhydride (20.63g, 89.79mmol) was added, and the reaction mixture was stirred at room temperature for 6 hours and concentrated in vacuo. The crude product was distributed between water (500mL) and EtOAc (200mL), and the aqueous layer was further extracted with EtOAc (2×100mL). The organic layers were combined and dried (Na 2 SO 4 The solvent was removed in vacuo to give (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (12.00 g, 72.3%) as a white solid. (LC / MS Method E): m / z 132 [M (-100) + H]+ (ES + ), at 1.50 min, UV active.

[0631] Step (ii): (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (0.5 g, 2.16 mmol) was dissolved in DMF (7 mL) and (R)-methyl 4-(1-aminoethyl)benzoate (0.46 g, 2.59 mmol) was added to the reaction mixture at room temperature. After that, HATU (1.23 g, 3.24 mmol) was added and the reaction mixture was stirred at room temperature for 30 minutes. After that, N,N-diisopropylethylamine (1.12 mL, 6.49 mmol) was added at room temperature and stirred at room temperature for 4 hours. The reaction mixture was concentrated 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% acetonitrile in water to give (R)-tert-butyl 3-(((R)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (0.6 g, 70.71%) as a white solid. (LC / MS Method E): m / z 293 [M (-100) + H] + (ES + ), at 1.81 minutes, UV active.

[0632] Step (iii): tert-Butyl (R)-3-(((R)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (0.5 g, 7.64 mmol) was dissolved in dioxane (3 mL) and the reaction was cooled to 0 °C. After this, 4N HCl in dioxane (5 mL) was added at room temperature and the reaction mixture was stirred at room temperature for 4 hours. The solvent was removed in vacuo and the crude material was purified by trituration with diethyl ether (5 mL) to give 4-((R)-1-((R)-morpholine-3-amido)ethyl)benzoate hydrochloride (0.4 g, 99.2%) as a white solid. (LC / MS Method E): m / z 293 [M+H] + (ES + ), at 0.87 min, UV active.

[0633] Step (iv): Methyl 4-((R)-1-((R)-morpholine-3-amido)ethyl)benzoate hydrochloride (0.3 g, 0.914 mmol) and DIPEA (0.47 mL, 2.70 mmol) were dissolved in acetonitrile (5 mL) at room temperature. After this, 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 6, 0.35 g, 1.095 mmol) was added and the reaction mixture was stirred at 80°C for 4 hours. The reaction mixture was concentrated 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 70% acetonitrile in water to give methyl 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.2 g, 41.2%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), at 1.23 minutes, UV active.

[0634] Step (v): Methyl 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoate (0.2 g, 0.376 mmol) was dissolved in dioxane (2 mL) and water (2 mL). Lithium hydroxide monohydrate (0.078 g, 1.88 mmol) was added and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was acidified with glacial acetic acid (1 mL) to adjust the pH to about 4 and concentrated in vacuo. The crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 30% acetonitrile in water to give 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (0.110 g, 56.5%) as a white solid.

[0635] 1H NMR: (400MHz, DMSO): δ (ppm) 1.36 (d, J = 6.8Hz, 3H), 2.15-2.10 (m, 1H), 2.24 (s, 3H), 2.74 (d, 1H, J = 12Hz), 2.99 (dd, 1H, J = 2.8Hz and J = 8.8Hz), 3.13 (d, 1H, J = 13.2Hz), 3.53-3.35 (m, 3H), 3.69-3.66 (m, 2H), 3.76 (d , 1H, J=8.4Hz), 2.99 (t, 1H, J=7.2Hz), 6.61 (s, 1H), 6.76 (d, 2H, J=15.2Hz), 7.22 (d, 1H, J=8.0Hz), 7.35 (d , 3H, J=8.0Hz), 7.73 (d, 1H, J=8.0Hz), 7.80-7.78 (m, 3H), 8.00 (s, 1H), 8.47 (d, 1H, J=8.0Hz), 9.67 (s, 1H).

[0636] (LC / MS method E): m / z 518 [M+H] + (ES + ), at 1.14 minutes, UV active.

[0637] The following compounds were prepared by following the procedures described in the previous examples but using different intermediates.

[0638] Example 2: 4-[(1S)-1-[[(3R)-4-[[6-(4-carbamoyl-2-methyl-phenyl)-2-pyridinyl]methyl]-morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 2)

[0639] Following the procedure described in Example 23, but using intermediates 1, 2, 15, the title compound was obtained. Note: Step (ii): Addition of additional catalyst, boronate and base was followed by an additional 1 hour of microwave irradiation.

[0640] HPLC method A in a gradient of 4% to 70% solvent B in solvent A. (LC / MS method C): m / z 503 [M+H] + (ES + ), at 1.74 minutes, UV active.

[0641] 1H NMR (DMSO, 400MHz): δ (ppm) 1.22 (d, J=7.0Hz, 3H), 2.27-2.35 (m, 4H), 2.68-2.76 (m, 1H), 3.10 (dd, J=9.3, 3.6Hz, 1H), 3.38-3.52 (m, 3H), 3.65-3.73 (m, 1H), 3.78 (dd, J=11.0, 3.6H z, 1H), 3.84 (d, J=13.9Hz, 1H), 4.88-4.97 (m, 1H), 7.21-7.28 (m, 2H), 7.33 (br.s, 1H), 7.3 8-7.48(m, 3H), 7.73-7.81(m, 4H), 7.81-7.90(m, 1H), 8.02(br.s, 1H), 8.48-8.55(m, 1H).

[0642] Example 3: 4-[(1S)-1-[[(3R)-4-[[2-(4-carbamoyl-2-methyl-phenyl)-4-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 3)

[0643] Following the procedure described in Example 1, but using intermediates 1, 2, 16, the title compound was obtained. Note: Step (ii): Addition of additional catalyst, boronate and base was followed by an additional 1 hour of microwave irradiation.

[0644] HPLC method B. (LC / MS method C): m / z 503 [M+H] + (ES + ) at 1.31 minutes, UV active.

[0645] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.32 (d, J=7.0Hz, 3H), 2.16-2.24 (m, 1H), 2.32 (s, 3H), 2.69-2.77 ( m, 1H), 3.04 (dd, J=8.8, 3.5Hz, 1H), 3.34 (d, J=14.3Hz, 1H), 3.47-3.61 (m, 2H), 3.63-3.71 (m, 1H ), 3.72-3.81(m, 2H), 4.89-4.98(m, 1H), 7.24-7.31(m, 2H), 7.31-7.37(m, 2H), 7.37-7.44(m, 1 H), 7.52 (brs, 1H), 7.73-7.83 (m, 4H), 8.04 (brs, 1H), 8.43 (d, J=8.1Hz, 1H), 8.55-8.59 (m, 1H).

[0646] Example 4: 4-[(1S)-1-[[(3R)-4-[[4-(4-carbamoyl-2-methyl-phenyl)-2-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 4)

[0647] Following the procedure described in Example 1, but using intermediates 1, 2, 17, the title compound was obtained.

[0648] HPLC method B. (LC / MS method C): m / z 503 [M+H] + (ES + ), at 1.51 minutes, UV active.

[0649] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.32 (d, J=6.9Hz, 3H), 2.24 (s, 3H), 2.26-2.34 (m, 1H), 2.69-2.75 (m, 1H), 3.10 (dd, J=9.3, 3.6Hz, 1H), 3.42-3.50 (m, 3H), 3.64-3.70 (m, 1H), 3.77 (dd, J=10.9, 3.6Hz, 1H), 3.85 ( d, J=14.1Hz, 1H), 4.89-4.97(m, 1H), 7.19-7.24(m, 2H), 7.26-7.31(m, 2H), 7.36(brs, 1H), 7.47-7.49( m, 1H), 7.74-7.81 (m, 3H), 7.81-7.84 (m, 1H), 8.12 (brs, 1H), 8.51 (d, J=8.1Hz, 1H), 8.54-8.58 (m, 1H).

[0650] Example 6: 6-[[[(3R)-4-[[3-(4-Carbamoyl-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]methyl]pyridine-3-carboxylic acid (Compound 6)

[0651] Following the procedure described in Example 1, but using intermediates 2, 18, 19, the title compound was obtained.

[0652] HPLC method C. (LC / MS method C): m / z 487 [M+H] - (ES - ), at 1.79 minutes, UV active.

[0653] 1 H NMR (CD 3OD, 400MHz): δ (ppm) 2.25-2.32 (m, 4H), 2.76-2.85 (m, 1H), 3.12 (dd, J=9.5, 3.7Hz, 1H), 3. 32-3.33(m, 1H), 3.54-3.68(m, 2H), 3.75-3.83(m, 1H), 3.95(dd, J=11.3, 3.7Hz, 1H), 4.02( d, J=13.0Hz, 1H), 4.48 (d, J=16.2Hz, 1H), 4.65 (d, J=16.2Hz, 1H), 7.20-7.26 (m, 2H), 7.32 -7.43(m, 4H), 7.67-7.75(m, 1H), 7.75-7.80(m, 1H), 8.16-8.24(m, 1H), 8.88-8.94(m, 1H).

[0654] Example 7: 4-((S)-1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 7)

[0655] Following the procedure described in Example 5, but using Intermediates 2 and 3, the title compound was obtained.

[0656] The product was purified by flash chromatography (reverse phase). (LC / MS method E): m / z 502 [M+H] + (ES + ), at 1.42 minutes, UV active.

[0657] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.38 (d, J=7.0Hz, 3H), 2.10-2.21 (m, 1H), 2.26 (s, 3H), 2.67 -2.75 (m, 1H), 2.91-3.06 (m, 1H), 3.25 (d, J=13.3Hz, 1H), 3.47-3.59 (m, 2H), 3.65-3.85 (m, 3H), 4.92-5.06 (m, 1H), 7.22-7.30 (m, 2H), 7.30-7.49 (m, 6H), 7.73-7.78 (m, 1H), 7. 82 (s, 1H), 7.85-7.90 (m, 2H), 7.99 (br.s, 1H), 8.56 (d, J=8.0Hz, 1H), 12.83 (br.s, 1H).

[0658] Example 8: 4-[[[(3R)-4-[[3-(4-Carbamoyl-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]methyl]-2,6-difluoro-benzoic acid (Compound 8)

[0659] Following the procedure described in Example 1, but using intermediates 2, 19, 20, the title compound was obtained. Note: Step (ii): Addition of additional catalyst, boronate and base was followed by an additional 2.5 hours of microwave irradiation.

[0660] HPLC method D. (LC / MS method C): m / z 524 [M+H] + (ES + ), at 1.83 minutes, UV active.

[0661] 1 H NMR (CD 3 OD, 400MHz): δ (ppm) 2.31 (s, 3H), 3.28-3.36 (m, 1H), 3.37-3.43 (m, 1H), 3.68-3.78 (m, 2H), 3.99-4.06 (m, 1H), 4.12 (dd, J=10.4, 4.0Hz, 1H), 4.18-4.27 (m, 2H), 4.45 (d, J=15 .5Hz, 1H), 4.51 (d, J=15.5Hz, 1H), 4.56 (d, J=12.8Hz, 1H), 7.00-7.06 (m, 2H), 7.30-7. 33(m, 1H), 7.47-7.54(m, 3H), 7.55-7.60(m, 1H), 7.73-7.78(m, 1H), 7.81-7.84(m, 1H).

[0662] Example 12: 4-[(1S)-1-[[(3R)-4-[[5-(2-methyl-4-sulfamoyl-phenyl)-3-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 12)

[0663] Following the procedure described in Example 1, but using Intermediates 1, 4, 21, the title compound was obtained.

[0664] HPLC method D. (LC / MS method D): m / z 539 [M+H] + (ES + ), at 1.78 minutes, UV active.

[0665] 1 H NMR (CD 3OD, 400MHz): δ (ppm) 1.49 (d, J=7.0Hz, 3H), 2.31 (s, 3H), 2.90-3.02 (m, 1H), 3.11-3.19 (m, 1H), 3.62-3.75 (m, 2H), 3.78 (dd, J=10.1, 3.3Hz, 1H), 3.91 -3.98(m, 2H), 4.10-4.19(m, 2H), 5.03-5.13(m, 1H), 7.39-7.48(m, 3H), 7.79-7.87(m, 1H), 7.86- 7.92 (m, 1H), 7.89-7.98 (m, 2H), 8.08-8.14 (m, 1H), 8.69 (d, J = 2.0Hz, 1H), 8.72 (d, J = 2.0Hz, 1H).

[0666] Example 13: 4-[[[(3R)-4-[[3-(4-Carbamoyl-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]methyl]-2-fluoro-benzoic acid (Compound 13)

[0667] Following the procedure described in Example 1, but using intermediates 2, 19, 22, the title compound was obtained.

[0668] HPLC method C. (LC / MS method D): m / z 506 [M+H] + (ES + ), at 2.31 minutes, UV active.

[0669] 1 H NMR (CD 3 OD, 400MHz): δ (ppm) 2.22-2.32 (m, 4H), 2.76-2.83 (m, 1H), 3.08 (dd, J=9.5, 3.6Hz, 1H), 3.2 4-3.31 (m, 1H), 3.54-3.65 (m, 2H), 3.73-3.80 (m, 1H), 3.85-3.93 (m, 2H), 4.32 (d, J=15.1Hz , 1H), 4.48 (d, J=15.1Hz, 1H), 6.98-7.03 (m, 1H), 7.03-7.08 (m, 1H), 7.19-7.27 (m, 2H), 7.2 9-7.34(m, 2H), 7.36-7.41(m, 1H), 7.57-7.64(m, 1H), 7.71-7.75(m, 1H), 7.77-7.80(m, 1H).

[0670] Example 14: 4-((S)-1-((R)-4-((4'-carbamoyl-2'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 14)

[0671] Following the procedure described in Example 5, but using intermediates 3 and 24, the title compound was obtained. Note: Step (iii): The compound was sufficiently pure after the workup; no chromatography was required.

[0672] The product was purified by acid / base extraction. (LC / MS Method E): m / z 504 [M+H] + (ES + ), at 1.16 minutes, UV active.

[0673] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.39 (d, J=7.0Hz, 3H), 2.06-2.21 (m, 1H), 2.69-2.77 (m, 1 H), 2.99 (dd, J=9.2, 3.5Hz, 1H), 3.21 (d, J=13.0Hz, 1H), 3.44-3.60 (m, 2H), 3.63-3.84 (m, 3H), 4.91-5.08 (m, 1H), 7.23-7.32 (m, 3H), 7.32-7.39 (m, 2H), 7.40-7.50 (m, 4H), 7 .52 (s, 1H), 7.82-7.99 (m, .3H), 8.51 (d, J=8.0Hz, 1H), 9.84 (br.s, 1H), 12.86 (s, 1H).

[0674] Example 16: 4-[(1S)-1-[[(3R)-4-[[2-(2-methyl-4-sulfamoyl-phenyl)-4-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 16)

[0675] Following the procedure described in Example 1, but using intermediates 1, 4, 16, the title compound was obtained. Note: Step (ii): Addition of additional catalyst and boronate was followed by an additional 1 hour of microwave irradiation.

[0676] HPLC method D. (LC / MS method D): m / z 539 [M+H] + (ES + ) at 1.98 minutes, UV active.

[0677] 1H NMR (DMSO, 400MHz): δ (ppm) 1.37 (d, J=7.0Hz, 3H), 2.35 (s, 3H), 2.70-2.83 (m, 1H) , 2.92-3.03(m, 1H), 3.54-3.69(m, 3H), 3.75-3.91(m, 2H), 3.98-4.10(m, 2H), 4.95 -5.04(m, 1H), 7.33-7.48(m, 5H), 7.54-7.58(m, 1H), 7.65(br.s, 1H), 7.71-7.75(m , 1H), 7.75-7.78(m, 1H), 7.85-7.90(m, 2H), 8.67-8.71(m, 1H), 8.95-9.03(m, 1H).

[0678] Example 17: 4-[(1S)-1-[[(3R)-4-[[4-(2-methyl-4-sulfamoyl-phenyl)-2-pyridinyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 17)

[0679] Following the procedure described in Example 1, but using intermediates 1, 4, 17, the title compound was obtained.

[0680] HPLC method D. (LC / MS method D): m / z 539 [M+H] + (ES + ), at 1.73 minutes, UV active.

[0681] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.38 (d, J=6.9Hz, 3H), 2.29 (s, 3H), 2.92-3.02 (m, 1H), 3.17-3.22 (m, 1H) , 3.63-3.75 (m, 2H), 3.80-3.93 (m, 2H), 4.03-4.10 (m, 1H), 4.13 (d, J = 13.9Hz, 1H), 4.24 (d, J = 13.9Hz, 1H), 4.96-5.05(m, 1H), 7.37-7.45(m, 3H), 7.45-7.48(m, 2H), 7.51-7.54(m, 1H), 7.54-7.58(m, 1H), 7 .72-7.76 (m, 1H), 7.78-7.81 (m, 1H), 7.84-7.89 (m, 2H), 8.72 (d, J = 5.1Hz, 1H), 9.11 (d, J = 7.7Hz, 1H).

[0682] Example 21: 2-Hydroxy-4-((S)-1-((R)-4-((5-(2-methyl-4-sulfamoylphenyl)pyridin-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 21)

[0683] Following the procedure described in Example 18, but using Intermediates 8 and 26, the title compound was obtained.

[0684] The product was purified by flash chromatography (reverse phase). (LC / MS method E): m / z 555 [M+H] + (ES + ), at 1.26 minutes, UV active.

[0685] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.33 (d, J=7.0Hz, 3H), 2.14-2.24 (m, 1H), 2.29 (s, 3H), 2.63-2.72 (m, 1 H), 3.04 (dd, J=9.2, 3.5Hz, 1H), 3.48-3.58 (m, 4H), 3.72-3.88 (m, 2H), 4.84-4.96 (m, 1H), 6.80 (d, J =8.0Hz, 1H), 6.83 (s, 1H), 7.40 (s, 2H), 7.44 (d, J = 8.0Hz, 1H), 7.66 (d, J = 8.0Hz, 1H), 7.72 (dd, J = 8 .0, 2.0Hz, 1H), 7.78 (d, J=2.0Hz, 1H), 7.82-7.89 (m, 1H), 8.49 (d, J=2.2Hz, 1H), 8.51-8.60 (m, 2H).

[0686] Example 22: 2-Hydroxy-4-((S)-1-((R)-4-((5-hydroxy-2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 22)

[0687] Following the procedure described in Example 18, but using Intermediates 8 and 27, the title compound was obtained.

[0688] The product was purified by flash chromatography (reverse phase). (LC / MS method E): m / z 570 [M+H] + (ES + ), at 1.29 minutes, UV active.

[0689] 1H NMR (DMSO, 400MHz): δ (ppm) 1.33 (d, J=7.0Hz, 3H), 2.10-2.20 (m, 1H), 2.27 (s, 3H), 2.67-2.76(m, 1H), 3.02(br.s, 1H), 3.11-3.21(m, 1H), 3.48-3.58(m, 2H), 3.66-3. 81(m, 3H), 4.84-4.94(m, 1H), 6.57-6.64(m, 1H), 6.75-6.88(m, 4H), 7.31-7.36(m, 3H), 7.63-7.70(m, 2H), 7.70-7.74(m, 1H), 8.47(d, J=7.9Hz, 1H), 9.58(br.s, 1H).

[0690] Example 24: 4-[(1S)-1-[[(3R)-4-[[4-hydroxy-3-(4-sulfamoylphenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 24)

[0691] Following the procedure described in Example 26, but using Intermediates 1 and 29, the title compound was obtained.

[0692] HPLC method D. (LC / MS method H): m / z 540 [M+H] + (ES + ) at 1.76 minutes, UV active.

[0693] 1 H NMR (CD 3 OD, 400MHz): δ (ppm) 1.51 (d, J = 7.0Hz, 3H), 3.25 (dd, J = 12.2, 3.9Hz, 1H), 3.36-3.44 ( m, 1H), 3.68-3.78 (m, 2H), 3.99-4.05 (m, 2H), 4.10 (d, J=12.8Hz, 1H), 4.19-4.29 (m, 2H ), 5.09-5.14 (m, 1H), 6.94 (d, J = 8.2Hz, 1H), 7.17 (dd, J = 8.2, 2.3Hz, 1H), 7.29 (d, J = 2. 3Hz, 1H), 7.47-7.51(m, 2H), 7.68-7.73(m, 2H), 7.89-7.94(m, 2H), 7.98-8.01(m, 2H).

[0694] Example 25: 2-Hydroxy-4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)benzoic acid (Compound 25)

[0695] Following the procedure described in Example 19, but using intermediates 8 and 4, the title compound was obtained. Note: Step (ii): 1.2 equivalents of the boronate were used. Step (iii): The reaction mixture was partitioned between EtOAc and 4N HCl; the organic layer was separated, concentrated, and the residue was purified by preparative HPLC.

[0696] HPLC method G. (LC / MS method E): m / z 554 [M+H] + (ES + ), at 1.68 minutes, UV active.

[0697] 1 H NMR (400MHz, DMSO) δ1.32 (d, J=7.0Hz, 3H), 2.10-2.21 (m, 1H), 2.27 (s, 3H), 2.63-2.76 (m, 1H), 3.01 (dd, J=9.2, 3.5Hz, 1H), 3.27 (d, J=13.2Hz, 1H), 3.45-3.58 (m, 2H), 3.65-3.82 (m, 3H), 4.81 -4.95(m, 1H), 6.67-6.71(m, 1H), 6.74(s, 1H), 7.23-7.28(m, 1H), 7.33-7.45(m, 6H), 7.59( d, J=8.0Hz, 1H), 7.69 (dd, J=8.0, 2.0Hz, 1H), 7.75 (d, J=2.0Hz, 1H), 8.40 (d, J=8.2Hz, 1H).

[0698] Example 27: 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)ethyl)-2,6-difluorobenzoic acid (single diastereomer of unknown stereochemistry 2, compound 27)

[0699] Following the procedure described in Example 29, but using intermediates 11 and 30, the title compound was obtained.

[0700] HPLC method F. (LC / MS method I): m / z 538 [M+H] + (ES + ), at 0.57 min, UV active.

[0701] 1 H NMR (CD 3OD, 400MHz): δ (ppm) 1.50 (d, J=7.0Hz, 3H), 2.29 (s, 3H), 3.22-3.27 (m, 1H), 3.35-3.41 (m, 1H), 3 .69-3.77 (m, 2H), 3.97-4.07 (m, 2H), 4.15 (d, J=12.8Hz, 1H), 4.26 (dd, J=12.7, 3.9Hz, 1H), 4.36 (d, J=12.8Hz, 1H), 5.07 (q, J=7.0Hz, 1H), 7.08-7.14 (m, 2H), 7.30 (d, J=8.0Hz, 1H), 7.39-7.44 ( m, 2H), 7.45-7.49 (m, 1H), 7.51-7.56 (m, 1H), 7.75 (dd, J=8.0, 1.8Hz, 1H), 7.82 (d, J=1.8Hz, 1H).

[0702] Example 28: 4-((S)-1-((R)-4-((5-(4-carbamoyl-2-methylphenyl)pyridin-3-yl)methyl)morpholine-3-amido)ethyl)-2-hydroxybenzoic acid (Compound 28)

[0703] Following the procedure described in Example 18, but using intermediates 8 and 31, the title compound was obtained.

[0704] HPLC method E. (LC / MS method E): m / z 519 [M+H] + (ES + ), at 1.36 minutes, UV active.

[0705] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.34 (d, J=7.0Hz, 3H), 2.13-2.24 (m, 1H), 2.26 (s, 3H), 2.63-2.76 (m, 1H) , 3.04 (dd, J=9.3, 3.5Hz, 1H), 3.33 (d, J=13.5Hz, 1H), 3.44-3.60 (m, 2H), 3.65-3.73 (m, 1H), 3.73-3. 88 (m, 2H), 4.83-4.96 (m, 1H), 6.78-6.87 (m, 2H), 7.32 (d, J=8.1Hz, 1H), 7.39 (br.s, 1H), 7.67 (d, J=8 .1Hz, 1H), 7.76-7.80(m, 1H), 7.82-7.88(m, 2H), 8.01(s, 1H), 8.45-8.50(m, 1H), 8.51-8.59(m, 2H).

[0706] Example 31: (R)-4-(1-(4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)cyclopropyl)benzoic acid (Compound 31)

[0707] Following the procedure described in Example 19, but using intermediates 2 and 32, the title compound was obtained. Note: Step (i): 3 equivalents of base were used. Step (ii): 1.5 equivalents of boronate were used.

[0708] The product was purified by flash chromatography (reverse phase). (LC / MS method E): m / z 514 [M+H] + (ES + ), at 1.46 minutes, UV active.

[0709] 1 H NMR (400MHz, DMSO) δ1.08-1.22(m, 2H), 1.22-1.32(m, 2H), 2.10-2.19(m, 1H), 2.26(s, 3H), 2.65- 2.76 (m, 1H), 2.98 (dd, J=9.3, 3.4Hz, 1H), 3.27 (d, J=13.4Hz, 1H), 3.47-3.54 (m, 1H), 3.55-3.62 (m , 1H), 3.67-3.73 (m, 1H), 3.79 (d, J=13.4Hz, 1H), 3.85 (dd, J=11.2, 3.4Hz, 1H), 7.15-7.22 (m, 2H), 7.23-7.29(m, 2H), 7.33-7.46(m, 4H), 7.73-7.84(m, 4H), 7.97(s, 1H), 8.89(s, 1H), 12.79(s, 1H).

[0710] Example 32: (R)-4-(1-(4-((4'-carbamoyl-2'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)cyclopropyl)benzoic acid (Compound 32)

[0711] Following the procedure described in Example 19, but using intermediates 24 and 32, the title compound was obtained. Note: Step (i): 3 equivalents of base were used. Step (ii): 1.5 equivalents of boronate were used.

[0712] The product was purified by flash chromatography (reverse phase). (LC / MS method E): m / z 516 [M+H] + (ES + ), at 1.45 minutes, UV active.

[0713] 1H NMR (400MHz, DMSO) δ1.29-1.49(m, 4H), 3.06-3.19(m, 1H), 3.55-3.73(m, 2H), 3.80-4.00(m, 2H), 4.13-4.49(m, 4H), 7.21-7.26(m, 2H), 7 .31-7.42(m, 4H), 7.46-7.53(m, 2H), 7.63-7.73(m, 2H), 7.83-7.89(m, 2H), 7.93(s, 1H), 9.95(s, 1H), 10.53(br.s, 1H), 12.85(br.s, 1H).

[0714] Example 33: (R)-4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)cyclopropyl)benzoic acid (Compound 33)

[0715]

[0716] Step (i) at room temperature, (R) -4- (1- (morpholine -3- amide) cyclopropyl) benzoic acid methyl ester, HCl salt (intermediate 32, 0.25g, 0.75mmol) and DIPEA (0.4mL, 2.26mmol) were dissolved in acetonitrile (3mL). 3'- (bromomethyl) -5'- hydroxy -2-methyl - [1,1'- biphenyl] -4- carboxamide (intermediate 6, 0.24g, 0.75mmol) was added, and the reaction mixture was stirred at 70 ° C for 2 hours. The reaction mixture was then distributed between water (70mL) and EtOAc (70mL). The aqueous layer was further extracted with EtOAc (2 × 25mL), and the organic layers were combined and dried (Na2O3). 2 SO 4 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 56% acetonitrile in water to give pure (R)-methyl 4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)cyclopropyl)benzoate (0.20 g, 49%) as a white solid. (LC / MS Method E): m / z 544 [M+H] + (ES + ), at 1.56 minutes, UV active.

[0717] Step (ii) (R)-methyl 4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)cyclopropyl)benzoate (0.20 g, 0.36 mmol) was dissolved in dioxane (2 mL) and water (1 mL). Lithium hydroxide (77 mg, 1.84 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was acidified to pH about 3 with glacial acetic acid (0.3 mL), and the reaction mixture was concentrated in vacuo. The crude product was purified by reverse phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product at 0% to 22% acetonitrile in water to give pure (R)-4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)cyclopropyl)benzoic acid (0.13 g, 66.7%) as an off-white solid. (LC / MS Method E): m / z 530 [M+H] + (ES + ), at 1.31 minutes, UV active.

[0718] 1 H NMR (DMSO, 400MHz): δ (ppm) 1.05-1.21 (m, 2H), 1.20-1.33 (m, 2H), 2.11-2.18 (m, 1H), 2.25 (s, 3H), 2.70-2.78 (m, 1H) ), 2.97 (dd, J=9.2, 3.5Hz, 1H), 3.17 (d, J=13.3Hz, 1H), 3.46-3.55 (m, 1H), 3.55-3.63 (m, 1H), 3.65-3.75 (m, 2H), 3. 83 (dd, J=10.9, 3.5Hz, 1H), 6.59-6.62 (m, 1H), 6.76-6.81 (m, 2H), 7.15-7.20 (m, 2H), 7.23 (d, J=7.9Hz, 1H), 7.33 (s , 1H), 7.72 (dd, J=7.9, 1.8Hz, 1H), 7.77-7.82 (m, 3H), 7.95 (s, 1H), 8.83 (s, 1H), 9.53 (br.s, 1H), 12.74 (br.s, 1H).

[0719] Example 34: (R)-4-(1-(4-((4'-carbamoyl-6-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-amido)cyclopropyl)benzoic acid (Compound 34)

[0720] Following the procedure described in Example 5, but using intermediates 33 and 34, the title compound was obtained. Note: steps (ii) and (iii) only.

[0721] The product was purified by flash chromatography (reverse phase). (LC / MS method E): m / z 516 [M+H] + (ES + ), at 1.07 minutes, UV active.

[0722] 1 H NMR (400MHz, DMSO) δ1.10-1.33 (m, 4H), 2.06-2.17 (m, 1H), 2.64-2.76 (m, 1H), 2.93 (dd, J=9.2, 3.5Hz, 1H) , 3.15 (d, J=12.8Hz, 1H), 3.45-3.59 (m, 2H), 3.62-3.73 (m, 2H), 3.83 (dd, J=10.9, 3.5Hz, 1H), 6.91 (d, J=8. 2Hz, 1H), 7.14 (dd, J=8.2, 2.2Hz, 1H), 7.17-7.21 (m, 2H), 7.23 (d, J=2.2Hz, 1H), 7.36 (s, 1H), 7.56-7.65 ( m, 2H), 7.78-7.85 (m, 2H), 7.86-7.92 (m, 2H), 7.99 (s, 1H), 8.86 (s, 1H), 9.67 (br.s, 1H), 12.52 (br.s, 1H).

[0723]

[0724]

[0725]

[0726]

[0727] Biological Activity

[0728] Cloning, baculovirus production, large-scale infection of HEK293 cells, and membrane preparation: Human prostaglandin E2 receptor 4 (EP 4 ) was cloned into the pBacMam expression vector (GeneScript, UK). EP was performed using the Invitrogen Bac-to-Bac baculovirus expression system 4Transposition of DNA. 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). Cells were incubated at 500 cm 3 The flask was seeded at a density of 3.5 million cells / mL in 5% v / v EP 4 Infection was performed under Bacman. Incubate at 37°C and 5% CO 2 Under, express in 36 hours period. Use PBS and cell scraper to remove cells. Cell culture is centrifuged at 2500RPM for 10 minutes at 4 ℃. Then pour out the supernatant, and store the precipitate at -80 ℃. The precipitate is thawed and resuspended in 15mL homogenizing buffer (20mM HEPES, 10mM EDTA, pH 7.4). Then homogenize in a mechanical homogenizer (VMR) for 10 seconds. The membrane is centrifuged at 40,000g for 15 minutes in a centrifuge tube at 4 ℃. The supernatant is poured out and resuspended in 15mL homogenizing buffer. Homogenize for 20 seconds. The membrane is centrifuged at 40,000g for 45 minutes at 4 ℃. The membrane is resuspended in 3mL mixed well storage buffer (20mM HEPES, 0.1mM EDTA, pH 7.4). The resulting membrane is then stored at -80 ℃.

[0729] cAMP Gs functional assay: The homogeneous time-resolved fluorescence (HTRF) cAMP dynamic-2 assay (Cisbio, France) was used to determine the expression of cAMP in EP 4 cAMP production after receptor activation HEK293 cells were transfected with 0.5% EP4Bacmam virus for 36 hours, then cells were dissociated and frozen at -150°C.

[0730] On the day of the test, increasing concentrations of test compounds were mixed with positive control (10 uM PGE 2 (Tocris, Abingdon, UK)) and a negative control (DMSO (Sigma-Aldrich, UK)) were added to a ProxiPlate-384 Plus, white 384-shallow well microplate (PerkinElmer, USA).

[0731] 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 6 The cell suspension was added to the pre-assigned assay plate using a multidrop, with a final assay concentration of 5000 cells / well. The plate was then incubated at 37°C and 5% CO 2 cAMP production was determined according to the manufacturer's instructions and the plates were read on a PheraStar fluorescence plate reader (BMG LabTech, Germany).

[0732] pEC was calculated using Dotmatics from the midpoint of the curve. 50 The values ​​(-Log of the molar concentration (M) in mol / L) are shown in Table 3.

[0733] Table 3: EP 4 pEC 50 and Emax value

[0734]

[0735]

[0736] One-way CACO-2 cell permeability assay

[0737] Caco-2 cells (ECACC) were cultured at 2×10 5 The cells were seeded into 24-well Transwell plates and incubated at 37°C with 5% CO 2 After 21 days of culture at 4 °C, the cells were used for confluent monolayers. The 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).

[0738] 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) including an analytical internal standard (0.5 μM carbamazepine).

[0739] Apparent permeability (P app) values ​​are shown in Table 4 and are determined according to the following relationship:

[0740]

[0741] 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 ).

[0742] Lucifer Yellow (LY) was added to the top buffer in all wells to assess the viability of the cell layer. Since LY cannot freely permeate 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.

[0743] Compounds recovered from the wells were determined based on the MS responses in the donor and acceptor compartments at the end of the incubation compared to the response in the donor compartment before the incubation (normalized to an internal standard).

[0744] Table 4 - Average Apparent Permeability (Papp) Values

[0745]

[0746]

[0747] It should be understood that although the invention has been described in conjunction with its detailed description, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the appended claims.

[0748] Numbered implementation plan

[0749] 1. A compound of formula I or a pharmaceutically acceptable salt or tautomer thereof:

[0750]

[0751] in:

[0752] A is OR', C(O)R', CO 2 R', C(O)N(R') 2 ,C(O)N(R')S(O) 2 R',S(O) 2 R',S(O) 2 OR', SO 2 N(R')2 , C 1-8 alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

[0753] Ring B is aryl or heteroaryl;

[0754] X and Y are each independently CR" or N, wherein at least one of X and Y is CH;

[0755] 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;

[0756] Each R 3 are independently selected from H, OR', COOR', C(O)R', halogen or C 1-6 alkyl;

[0757] R 4 It is H, C 1-6 Alkyl, halogen, CN, NO 2 、OR'、CO 2 R' or C(O)R';

[0758] R 5 is OR', C(O)R', CO 2 R', C(O)N(R') 2 ,C(O)N(R')S(O) 2 R',S(O) 2 R',S(O) 2 OR', OSO 2 N(R') 2 ;

[0759] Each R' is independently H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0760] Each R" is H, C 1-6 alkyl, halogen or OR'; and

[0761] n is 0, 1, 2, or 3;

[0762] wherein at each occurrence, alkyl and cycloalkyl are each optionally and independently replaced by OH, SH, CN, NO 2 , CO 2 H, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group;

[0763] wherein heterocycloalkyl, aryl and heteroaryl are each optionally and independently substituted by OR', SR', CN, NO 2 , CO 2 R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0764] 2. The 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, NO 2 , CO 2 R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

[0765] 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.

[0766] 4. A compound of formula I, which is a compound of formula (1) or a pharmaceutically acceptable salt or tautomer thereof:

[0767]

[0768] in:

[0769] A is selected from:

[0770]

[0771] 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; and

[0772] X and Y are each independently selected from CH, CF, COH or N;

[0773] 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;

[0774] R 3 is H, OH or F;

[0775] R 4 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 atoms1-3 alkyl;

[0776] R 5 OH, CO 2 H.CONH 2 、SO 2 NH 2 or OSO 2 NH 2 ;and

[0777] R 6 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring.

[0778] 5. A compound according to embodiment 4, wherein A is:

[0779]

[0780] 6. The compound of embodiment 4, which is a compound of formula (2a) or formula (2b) or a pharmaceutically acceptable salt thereof:

[0781]

[0782] Among them U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 and R 5 Same as defined in Embodiment 4.

[0783] 7. A compound according to any one of embodiments 1 to 6, wherein R 1 is H or methyl or R 1 With R 2 Connect to form a cyclopropane-1,1-diyl ring.

[0784] 8. The compound according to embodiment 7, wherein R 1 It's methyl.

[0785] 9. A compound according to any one of embodiments 1 to 6, wherein R 2 It's H.

[0786] 10. The compound according to Embodiment 4, which is a compound of formula (3a) or (3b) or a pharmaceutically acceptable salt thereof:

[0787]

[0788] Among them U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4and R 5 Same as defined in Embodiment 4.

[0789] 11. A compound according to any one of embodiments 1 to 10, wherein R 3 It's H.

[0790] 12. A compound according to any one of embodiments 1 to 11, wherein R 4 It is H, OH or methyl.

[0791] 13. The compound according to embodiment 12, wherein R 4 It's methyl.

[0792] 14. A compound according to any one of embodiments 1 to 13, wherein R 5 It is CONH 2 or SO 2 NH 2 .

[0793] 15. A compound according to any one of embodiments 4 to 14, wherein X and Y are both CH.

[0794] 16. A compound according to any one of embodiments 4 to 15, wherein U, V, W and Z are CH.

[0795] 17. A compound according to any one of embodiments 4 to 15, wherein U, V and Z are CH, and W is COH.

[0796] 18. A compound according to embodiment 1, selected from:

[0797]

[0798]

[0799]

[0800]

[0801] or a pharmaceutically acceptable salt thereof.

[0802] 19. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 18 and a pharmaceutically acceptable excipient.

[0803] 20. A compound according to any one of embodiments 1 to 18 or a composition according to embodiment 19 for use in the treatment of an EP4 receptor mediated disease.

[0804] 21. The compound or composition for use according to embodiment 20, wherein the EP4 receptor-mediated disease is a gastrointestinal disorder.

[0805] 22. The compound or composition for use according to embodiment 21, wherein the gastrointestinal disorder is selected from constipation disorder, 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.

[0806] 23. The compound or composition for use according to embodiment 20, wherein the EP4 receptor mediated disease is a lung disease and disorder.

[0807] 24. The compound or composition for use according to embodiment 23, 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, C optionally substituted by 1 to 3 fluorine atoms 1-3 Alkoxy, halogen, CN, NO2, OR', CO2R' or C(O)R'; R 5 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 is 0, 1, 2, or 3; wherein at each occurrence, alkyl and cycloalkyl are each optionally and independently substituted with OH, SH, CN, NO2, CO2H, halogen or COOC 1-4 Up to 3 instances of substitution in the alkyl group; wherein 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 5- to 6-membered heteroaryl, each of which is optionally and independently substituted by OR', SR', CN, NO2, CO2R', halogen, C 1-4 Up to 3 instances of alkyl or oxo are substituted.

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, wherein n is 0, 1 or 2, preferably wherein n is 0 or 1.

5. The compound according to any one of claims 1 to 4, 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.

6. The compound according to any one of claims 1 to 5, wherein A is selected from: and R 6 It is C 1-3 Alkyl or C 3-6 Cycloalkyl ring; preferably wherein A is: More preferably wherein A is 7. The compound according to claim 5, which is a compound of formula (2a) or formula (2b) 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 and R 5 Same as defined in claim 5.

8. A compound according to any one of claims 1 to 7, wherein R 1 and R 2 are each independently selected from 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.

9. A compound according to any one of claims 1 to 8, wherein R 1 is H or methyl, or R 1 With R 2 connected to form a cyclopropane-1,1-diyl ring; preferably wherein R 1 It's methyl.

10. A compound according to any one of claims 1 to 9, wherein R 2 It's H.

11. The compound according to claim 5, which is a compound of formula (3a) or formula (3b) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or optical isomer thereof: Among them U, V, W, X, Y, Z, R 3 , R 4 and R 5 Same as defined in claim 5.

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

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

14. A compound according to any one of claims 1 to 13, wherein R 5 is selected from OH, CO2H, CONH2, SO2NH2 or OSO2NH2; preferably wherein R 5 is OH, CONH2, SO2NH2 or OSO2NH2; more preferably wherein R 5 It is CONH2 or SO2NH2.

15. A compound according to any one of claims 1 to 14, wherein X and Y are each independently selected from CH, CF, COH or N; preferably wherein X is CH or N, and Y is selected from CH, CF, COH or N; more preferably wherein both X and Y are CH.

16. A compound according to any one of claims 5 to 15, wherein: (i) U is CH or N; and / or (ii) V is CH or N; and / or (iii) W is selected from CH, COH, N or N + -O - ; and / or (iv) Z is selected from CH, COH or N.

17. A compound according to any one of claims 5 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, which 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 as a medicament.

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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