Morpholine-3-carboxamide derivatives as prostaglandin E2 receptor 4 (EP4) agonists for the treatment of gastrointestinal and pulmonary diseases

Abstract

The present invention relates to compounds of formula (1) as prostaglandin E2 receptor 4 (EP4) agonists for use in methods for the treatment of gastrointestinal and pulmonary diseases or disorders. The disclosure provides exemplary compounds and pharmacological data. JPEG2025525852000098.jpg45170

Description

[Technical Field]

[0001] This application relates to novel compounds and their use as prostaglandin E2 receptor 4 (EP4) agonists. The compounds described herein may be useful in the treatment or prevention of diseases in which the EP4 receptor is involved. This application is also directed to pharmaceutical compositions containing these compounds, as well as the manufacture and use of these compounds and compositions in the prevention or treatment of diseases in which the EP4 receptor is involved. [Background technology]

[0002] Prostanoids, including prostaglandins and thromboxanes, are metabolite derivatives of arachidonic acid that play important roles in cellular physiology. Arachidonic acid is an essential component of membrane phospholipids, liberated via the activity of phospholipase A2 (PLA2). Prostaglandin biosynthesis is mediated by cyclooxygenase (COX), which converts arachidonic acid to an unstable intermediate (PGH2), generating prostaglandins, including PGE2. PGE2 is the most widely produced prostanoid, whose activity is mediated through action at four functionally distinct receptor subtypes, EP1-EP4.

[0003] EP receptors belong to the G protein-coupled receptor (GPCR) family, which are integral membrane proteins with seven transmembrane domains. This receptor class can be broadly categorized according to their signaling pathways: i) Gs-coupled receptors (activating adenylate cyclase and producing cyclic adenosine monophosphate (cAMP)) EP2 and EP4, ii) Gq-coupled receptors (activating PLC) EP1, and iii) Gi-coupled receptors (inhibiting adenylate cyclase) EP3.

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

[0005] The EP4 receptor plays an important role in various physiological functions, including gastrointestinal homeostasis, regulation of vascular tone, renal function, inflammation, fever, and carcinogenesis. The potent biological actions 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 disorders (FGIDs), including chronic constipation, are common gastrointestinal conditions encountered by primary care physicians and gastroenterologists. The prevalence of chronic constipation ranges from 1% to 8%, and it can negatively impact quality of life (QoL) and impose a significant social and economic burden. Chronic constipation can cause discomfort and affect patients' daily lives. Symptoms include hard, lumpy stools, straining during defecation, and a feeling of incomplete evacuation. The use of laxatives and stool softeners remains common, despite many patients not achieving adequate relief. There remains a need for treatments that can provide complete and sustainable symptom relief.

[0007] The intestinal mucosa plays an important role in anion homeostasis and fluid maintenance. These functions are mediated through coordinated ion transport via membrane-bound transporters and channels located on the apical and basolateral membranes of intestinal epithelial cells. PGE2 is a known secretagogue that can directly promote chloride ion secretion from intestinal epithelial cells. The secretory action of PGE2 is mediated in part through the EP4 receptor, which can stimulate chloride anion secretion across the intestinal mucosa. Lubiprostone, a bicyclic fatty acid derivative of PGE1 clinically approved for the treatment of chronic constipation and IBS-C, has been shown to promote fluid secretion and gastrointestinal motility through activation of prostaglandin (EP4) receptors. EP4-selective agonists may be of therapeutic value in promoting intestinal fluid homeostasis in chronic constipation.

[0008] Inflammatory bowel disease (IBD) is a chronic, debilitating gastrointestinal disorder that includes ulcerative colitis and Crohn's disease. Patients with IBD typically experience symptoms including diarrhea, abdominal pain, weight loss, rectal bleeding, and fever. Clinical management involves strategies to control abnormal, dysregulated immune responses in the intestinal mucosa. Depending on the severity of the disease, a wide range of medications are used to induce and maintain remission. These medications include aminosalicylates, corticosteroids, immunosuppressants, antibiotics, and biologic agents. This may be done in a stepwise approach, with treatment intensification depending on the severity and progression of the disease. However, it is clear that some patients are refractory to drug therapy or cannot tolerate it due to systemic side effects. With current treatment strategies, only a small proportion of patients achieve long-term remission, suggesting the need for improved treatment.

[0009] Intestinal barrier dysfunction plays an important pathogenic role in IBD, and there is growing interest in developing drugs that restore barrier function (mucosal healing). EP4 is expressed in several cell types, including gastrointestinal (GI) epithelial cells, lamina propria mononuclear cells, and colonic innervating sensory neurons, and may provide benefits by protecting the GI mucosal barrier in addition to suppressing inappropriate mucosal immune responses. EP4-mediated PGE2 signaling promotes cell differentiation into wound-associated epithelial cell phenotypes, which are important for wound repair (Miyoshi, H. et al. EMBO J. 2017, 36, 5-24). Administration of an EP4 agonist is effective in chemically induced colitis models (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), whereas EP4-deficient mutant mice 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). A small Phase II study evaluated the EP4 agonist ONO-4819CD 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 was not powered for efficacy, patients treated with ONO-4819CD showed signs of improvement in Disease Activity Index (DAI) scores and histological scores. These data support the potential clinical benefit of EP4 agonist therapy in IBD.

[0010] Asthma and chronic obstructive pulmonary disease are inflammatory airway disorders characterized by airway limitation. It is estimated that approximately 300 million people have asthma, making it the most common chronic disease in children. Despite advances in asthma management, a significant proportion of patients experience uncontrolled disease, which may lead to mortality and morbidity. There remains a need for treatments that can achieve effective control of asthma symptoms and reduce the risk of future exacerbations with long-term management. PGE2 is known to have bronchodilatory and anti-inflammatory effects in isolated airway smooth muscle of rodents and humans. Inhaled PGE2 has been shown to be beneficial for inflammation and airway dilation in patients with chronic bronchitis and asthma. However, PGE2 also induces reflex coughing, likely via stimulation of the upper airway via the EP3 receptor. This has led to ongoing efforts to discover EP receptor-selective drugs for the treatment of airway disorders. Interestingly, significant interspecies differences have been reported in the receptor subtypes that mediate airway smooth muscle relaxation. In guinea pigs, monkeys, and mice, EP2 agonists can induce relaxation of airway smooth muscle, while in humans, relaxation of airway smooth muscle is mediated by EP4 receptors (Buckley, J. et al. Thorax 2011, 66, 1029-1035). Selective EP4 receptor agonists may have promising therapeutic effects in airway diseases.

[0011] The EP4 receptor has been shown to play a role in blood pressure regulation. EP4 is expressed on smooth muscle cells and endothelial cells and can induce vasodilatory effects through endothelial nitric oxide synthase (eNOS)-mediated nitric oxide (NO) production. PGE2 has been shown to dose-dependently relax smooth muscle in aortic rings, and this effect is abolished in EP4 knockout mice. In halothane-anesthetized dogs, the EP4-selective agonist ONO-AE1-329 induced a vasopressor response (Honda, A. et al. Eur. J. Pharmacol. 2016, 775, 130-137). Similarly, hypotension was reported as a side effect in IBD patients treated with ONO-4819CD (Nakase, H. et al. Inflamm. Bowel Dis. 2010, 16, 731-733).

[0012] EP4 agonists that can be used to treat various gastrointestinal disorders and airway disorders without causing systemic side effects on the cardiovascular system may have promising therapeutic effects.In particular, EP4 agonists that can be used to treat various gastrointestinal disorders without causing systemic side effects on the cardiovascular system may have promising therapeutic effects.There is a need to discover safe and effective EP4 selective drugs. Summary of the Invention

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

[0014] As used herein, a compound of formula I: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; 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 1-8alkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; Ring B is aryl or heteroaryl; X and Y are each, independently of each other, CR″ or N, and at least one of X and Y is CH; R 1 and R 2 are, independently of each other, H, C 1-6 Alkyl, C 1-6 Alkoxy or R 1 and R 2 together with the carbon atoms to which they are attached, form C 3-6 forming cycloalkane-1,1-diyl; Each R 3 are, independently of each other, H, OR', COOR', C(O)R', halo, and C 1-6 alkyl; R 4 is H, C 1-6 Alkyl, C optionally substituted with 1 to 3 fluorine atoms 1-3 alkoxy, halo, CN, NO2, OR', CO2R', or C(O)R'; R 5 is OR', OC(O)R', OC(O)OR', COR', CON(R')2, SON(R')2, SOR', OSOR', or OSON(R')2; Each R' is independently H, C 1-6 Alkyl or C 3-6 is cycloalkyl; Each R" is H, C 1-6 alkyl, halo, or OR'; n is 0, 1, 2, or 3; Each occurrence of alkyl and cycloalkyl may optionally, and independently of one another, be selected from up to three of OH, SH, CN, NO, COH, halo, or COOC. 1-4 is substituted with alkyl; Heterocycloalkyl, aryl, and heteroaryl each optionally, and independently of one another, may be selected from up to three of OR', SR', CN, NO2, CO2R', halo, C 1-4 substituted with alkyl or oxo; Provided are compounds of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof:

[0015] In another aspect, the present invention includes pharmaceutical compositions comprising a compound described herein and a pharmaceutically acceptable excipient.

[0016] 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 aminosalicylates, corticosteroids, immunomodulators, and combinations thereof.

[0017] In another aspect, the invention includes a compound described herein, a composition described herein, or a kit described herein for use as a medicament.

[0018] 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.

[0019] In another aspect, the present invention includes a method for modulating EP4 receptor agonist activity in a biological sample, comprising contacting the EP4 receptor with a compound or composition described herein.

[0020] In another aspect, the present invention includes a method for treating an EP4 receptor-mediated disease, comprising administering to a patient in need of treatment for an EP4 receptor-mediated disease a compound or composition described herein.

[0021] The above-mentioned compounds can be used herein as EP4 receptor agonists.The above-mentioned compounds can act selectively at the EP4 receptor.The above-mentioned compounds can be used in the manufacture of compositions or medicaments.The compounds, compositions, or medicaments can be used to treat, prevent, improve, control, or reduce the risk of diseases or disorders involving the EP4 receptor. The compounds, compositions, or medicaments described above may be used to treat, prevent, ameliorate, control, or reduce the risk of gastrointestinal disorders and conditions including, but not limited to, constipation, 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 dysfunction, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrhea, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis.

[0022] The compounds, compositions, or medicaments described above 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 idiopathic cough, reactive airways disease, and idiopathic pulmonary fibrosis. DETAILED DESCRIPTION OF THE INVENTION

[0023] The present invention relates to novel compounds. The present invention also relates to the use of the novel compounds as agonists of the EP4 receptor. The present invention further relates to the use of the novel compounds in the manufacture of medicaments for use as EP4 receptor agonists, and to methods of treatment comprising administering the compounds of the present invention as EP4 receptor agonists.

[0024] The compounds of formula I can be used to treat, prevent, ameliorate, control, or reduce the risk of diseases or disorders in which the EP4 receptor is involved. The compounds of Formula I, such compounds, compositions, or medicaments, can be used to treat, prevent, ameliorate, control, or reduce the risk of gastrointestinal disorders and conditions including, but not limited to, constipation, 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 dysfunction, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrhea, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis.

[0025] 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 idiopathic cough, reactive airways disease, and idiopathic pulmonary fibrosis.

[0026] Certain novel compounds of the present invention exhibit particularly high activity as EP4 receptor agonists.

[0027] The compounds of the present invention have been shown to have activity as EP4 receptor agonists.The compounds of the present invention also have low gastrointestinal permeability, as shown by Caco-2 studies.Therefore, it is believed that the compounds of the present invention have low systemic bioavailability when orally administered.The agonistic effect on the function of EP4 receptors expressed in the gastrointestinal tract may provide treatment for various gastrointestinal disorders.The combination of EP4 receptor agonist activity and low gastrointestinal permeability suggests that the compounds of the present invention are useful for treating various gastrointestinal disorders without causing adverse cardiovascular effects resulting from systemic distribution.

[0028] In one embodiment, the present invention provides a compound of formula I: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; 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 of each other, CR″ or N, and at least one of X and Y is CH; R 1 and R 2 are, independently of each other, H, C 1-6 Alkyl, C 1-6 Alkoxy or R 1 and R 2 together with the carbon atoms to which they are attached, form C 3-6 forming cycloalkane-1,1-diyl; Each R 3 are, independently of each other, H, OR', COOR', C(O)R', halo, and C 1-6 alkyl; R 4 is H, C 1-6 Alkyl, C optionally substituted with 1 to 3 fluorine atoms 1-3 alkoxy, halo, CN, NO2, OR', CO2R', or C(O)R'; R 5 is OR', OC(O)R', OC(O)OR', COR', CON(R')2, SON(R')2, SOR', OSOR', or OSON(R')2; Each R' is independently H, C 1-6 Alkyl or C 3-6 is cycloalkyl; Each R" is H, C 1-6 alkyl, halo, or OR'; n is 0, 1, 2, or 3; Each occurrence of alkyl and cycloalkyl may optionally, and independently of one another, be selected from up to three of OH, SH, CN, NO, COH, halo, or COOC. 1-4 is alkyl substituted; Heterocycloalkyl, aryl, and heteroaryl each optionally, and independently of one another, may be selected from up to three of OR', SR', CN, NO2, CO2R', halo, C 1-4 substituted with alkyl or oxo; Provided are compounds of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof:

[0029] In some embodiments, the compound is a compound of formula I: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; 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 of each other, CR″ or N, and at least one of X and Y is CH; R 1 and R 2 are, independently of each other, H, C 1-6 Alkyl, C 1-6 Alkoxy or R 1 and R 2 together with the carbon atoms to which they are attached, form C 3-6 forming a cycloalkane-1,1-diyl ring; Each R 3are, independently of each other, H, OR', COOR', C(O)R', halo, and C 1-6 alkyl; R 4 is H, C 1-6 alkyl, halo, CN, NO2, OR', CO2R', or C(O)R'; R 5 is OR', OC(O)R', OC(O)OR', COR', CON(R')2, SON(R')2, SOR', OSOR', or OSON(R')2; Each R' is independently H, C 1-6 Alkyl or C 3-6 is cycloalkyl; Each R" is H, C 1-6 alkyl, halo, or OR'; n is 0, 1, 2, or 3; Each occurrence of alkyl and cycloalkyl may optionally, and independently of one another, be selected from up to three of OH, SH, CN, NO, COH, halo, or COOC. 1-4 is alkyl substituted; Heterocycloalkyl, aryl, and heteroaryl each optionally, and independently of one another, may be selected from up to three of OR', SR', CN, NO2, CO2R', halo, C 1-4 substituted with alkyl or oxo; A compound of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof:

[0030] In some embodiments, the compound is a compound of Formula I, Formula (1), Formula (1a), Formula (1b), Formula (2), Formula (2a), Formula (2b), Formula 3, Formula (3a), Formula (3b), Formula (4), Formula (4a), Formula (4b), Formula IIa, IIb, IIc, IId, or IIe, or a pharmaceutically acceptable salt or tautomer thereof.

[0031] In some embodiments, the compound is a compound of Formula I, Formula (1), Formula (1a), Formula (1b), Formula (2), Formula (2a), Formula (2b), Formula 3, Formula (3a), Formula (3b), Formula (4), Formula (4a), Formula (4b), Formula IIa, IIb, IIc, IId, or IIe, or a pharmaceutically acceptable salt thereof.

[0032] In some embodiments, a compound of formula IIa, a compound of formula IIb, a compound of formula IIc, a compound of formula IId, or a compound of formula IIe: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof, A, X, Y, R 1 , R 2 , R 3 , R 4 , and R 5 is as defined above, A compound of formula IIa, a compound of formula IIb, a compound of formula IIc, a compound of formula IId, or a compound of formula IIe, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

[0033] In some embodiments, A is CO2R', C(O)N(R')S(O)2R', S(O)2R', or heteroaryl.

[0034] In some embodiments, A is COOH or 5-membered heteroaryl optionally substituted with OR' or SR'.

[0035] In some embodiments, ring B may each optionally, and independently of one another, be selected from the group consisting of up to three of OR', SR', CN, NO, COR', halo, C 1-4 It is a 5- or 6-membered aryl or 5- or 6-membered heteroaryl substituted with alkyl or oxo.

[0036] In some embodiments, Ring B is a 5-6 membered aryl or a 5-6 membered heteroaryl.

[0037] In some embodiments, Ring B is phenyl optionally substituted with up to 3 OH.

[0038] In some embodiments, Ring B is a 6-membered heteroaryl containing 1 or 2 nitrogen atoms, each of which is optionally substituted with oxo.

[0039] In some embodiments, Ring B is a phenyl, pyridine, or pyridine-N-oxide ring, each optionally substituted with OR'.

[0040] In some embodiments, each R 3 are, independently of each other, OR', halo, or C 1-6 alkyl.

[0041] In some embodiments, each R 3 are independently selected from OR′ or alkyl; n is 1.

[0042] In some embodiments, n is 0, 1, or 2.

[0043] In some embodiments, n is 0 or 1.

[0044] In some embodiments, n is 0.

[0045] In some embodiments, R 4 is H, C 1-6 It is alkyl, halo, or OR'.

[0046] In some embodiments, R 4 is H, methyl, or OH.

[0047] In some embodiments, R5 is OR', OC(O)R', CO2R', CON(R')2, SO2N(R')2, SO2R', or OSO2N(R')2.

[0048] In some embodiments, R 5 is OR', CO2R', CON(R')2, SO2N(R')2, or OSO2N(R')2.

[0049] In some embodiments, R 5 is OH, CO2H, CONH2, SO2NH2, or OSO2NH2.

[0050] In some embodiments, R 5 is CONH2 or SO2NH2.

[0051] In some embodiments, R 5 is CONH2.

[0052] In some embodiments, the compound is a compound of formula (1): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof, U, V, W, and Z are each independently CH, COH, N, or N + -O - is selected from the group consisting of at least three of U, V, W, and Z are CH; X, Y, R 1 , R 2 , R 3 , R 4 , and R 5 is as described herein, A compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

[0053] In some embodiments, the compound is a compound of formula (1): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; A is, [ka] selected from the group consisting of: U, V, W, and Z are each independently CH, COH, N, or N + -O - wherein at least three of U, V, W, and Z are CH; X and Y are each, independently of one another, selected from the group consisting of CH, CF, COH, or N; R 1 and R 2 are each independently H, C optionally substituted with 1 to 3 fluorine atoms, 1-3 alkyl or R 2 C optionally substituted with 1 to 3 fluorine atoms by bonding to 3-6 forming a cycloalkyl ring; R 3 is H, OH, or F; R 4 is H, OH, CN, halo, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkoxy or C optionally substituted with 1 to 3 fluorine atoms 1-3 is alkyl; R 5 is OH, CO2H, CONH2, SO2NH2, or OSO2NH2; R 6 is C 1-3 Alkyl or C 3-6 is a cycloalkyl ring, A compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof:

[0054] In some embodiments, A is [ka] selected from the group consisting of: R 6 is C 1-3 Alkyl or C 3-6 It is a cycloalkyl ring.

[0055] In some embodiments, A is [ka] is.

[0056] In some embodiments, R 3 is halo or OR'. In another embodiment, R 3 is F or OH, and n is 0, 1, or 2.

[0057] In some embodiments, X and Y are each, independently of each other, selected from the group consisting of CH, CF, COH, or N.

[0058] In some embodiments, at least one of X and Y is CH.

[0059] In some embodiments, X and Y are both CH.

[0060] In some embodiments, provided herein are compounds of formula IIa, IIb, IIc, IId, or IIe.

[0061] In some embodiments, R 1 and R 2 are, independently of each other, H, C 1-6 alkyl, halo, or hydroxy, or R 1 and R 2 together with the carbon atoms to which they are attached, form C 3-6 Members (C 3-6(membered) form a cycloalkyl.

[0062] In some embodiments, R 1 and R 2 are each independently selected from H, C optionally substituted with 1 to 3 fluorine atoms, 1-3 alkyl, or R 1 is R 2 C optionally substituted with 1 to 3 fluorine atoms by bonding to 3-6 It forms a cycloalkyl ring.

[0063] In some embodiments, R 1 and R 2 are each, independently of one another, H or methyl, or R 1 and R 2 together with the carbon atom to which they are attached form cyclopropyl.

[0064] In some embodiments, R 4 H, OH, halo, C 1-6 Alkoxy or C 1-6 In another embodiment, R is an alkyl, wherein each of the alkyl and alkoxy is optionally, and independently of each other, substituted with halo. 4 is H, OH, or methyl.

[0065] In some embodiments, R 5 is OR', CO2R', CON(R')2, SO2N(R')2, or OSO2N(R')2. In another embodiment, R 5 is OH, CO2H, CONH2, SO2NH2, or OSO2NH2.

[0066] In some embodiments, the present invention includes a pharmaceutical composition comprising a compound described herein, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and a pharmaceutically acceptable excipient.

[0067] In some embodiments, the present invention includes a pharmaceutical composition comprising a compound or salt described herein and a pharmaceutically acceptable excipient.

[0068] In some embodiments, the pharmaceutical composition further comprises at least one additional therapeutic agent selected from the group consisting of an aminosalicylates, corticosteroids, immunomodulators, and combinations thereof.

[0069] In another aspect, the present invention includes a method for modulating EP4 receptor agonist activity in a biological sample, comprising contacting the EP4 receptor with a compound or salt or composition described herein.

[0070] In yet another aspect, the present invention includes a method for treating an EP4 receptor-mediated disease, comprising administering to a patient in need of treatment for an EP4 receptor-mediated disease a compound or salt or composition described herein.

[0071] In one embodiment of this aspect, the EP4 receptor-mediated disease is a gastrointestinal disorder.

[0072] In another embodiment, the gastrointestinal disorder is selected from the group consisting of constipation, 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 dysfunction, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrhea, immune-mediated gastrointestinal disorders, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis, or pulmonary diseases and conditions such as chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, reactive airways disease, and idiopathic pulmonary fibrosis.

[0073] In some embodiments, the compound of Formula (1a) or Formula (1b): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof, U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , and R 5 is as described herein, Provided herein are compounds of Formula (1a) or Formula (1b), or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof:

[0074] In some embodiments, a compound of Formula (2), Formula (2a), or Formula (2b): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof, A, U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , and R 5 is as described herein, Provided herein are compounds of Formula (2), Formula (2a), or Formula (2b), or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

[0075] In some embodiments, the compound of Formula (2a) or Formula (2b): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof; U, V, W, and Z are each independently CH, COH, N, or N + -O - wherein at least three of U, V, W, and Z are CH; X and Y are each, independently of one another, selected from the group consisting of CH, CF, COH, or N; R 1 and R 2 are each independently H, C optionally substituted with 1 to 3 fluorine atoms, 1-3 alkyl or R 2 C optionally substituted with 1 to 3 fluorine atoms by bonding to 3-6 forming a cycloalkyl ring; R 3 is H, OH, or F; R 4 is H, OH, CN, halo, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkoxy or C optionally substituted with 1 to 3 fluorine atoms 1-3 is alkyl; R 5 is OH, CO2H, CONH2, SO2NH2, or OSO2NH2, Provided herein are compounds of Formula (2a) or Formula (2b), or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof:

[0076] In some embodiments, a compound of Formula (3), Formula (3a), or Formula (3b): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof, A, U, V, W, X, Y, Z, R 3 , R 4 , and R 5 is as described herein, Provided herein are compounds of Formula (3), Formula (3a), or Formula (3b), or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof:

[0077] In some embodiments, the compound of Formula (3a) or Formula (3b): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof; U, V, W, and Z are each independently CH, COH, N, or N + -O - wherein at least three of U, V, W, and Z are CH; X and Y are each, independently of one another, selected from the group consisting of CH, CF, COH, or N; R 3 is H, OH, or F; R 4 is H, OH, CN, halo, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkoxy or C optionally substituted with 1 to 3 fluorine atoms 1-3 is alkyl; R 5 is OH, CO2H, CONH2, SO2NH2, or OSO2NH2, Provided herein are compounds of Formula (3a) or Formula (3b), or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof:

[0078] In some embodiments, a compound of Formula (4), Formula (4a), or Formula (4b): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof, A, U, V, W, X, Y, Z, R 3 , R 4 , and R 5 is as described herein, Provided herein are compounds of formula (4), formula (4a), or formula (4b).

[0079] In some embodiments, the compound is a compound of formula (1): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; A is, [ka] and; U is CH or N; V is CH or N; W is CH, COH, N, or N + -O - selected from the group consisting of: Z is selected from the group consisting of CH, COH, or N; X is CH or N; Y is CH, CF, COH, or N; R 1 and R 2 are each independently selected from the group consisting of H or methyl, or R 1 and R 2 together with the carbon atom to which they are attached form a cyclopropyl; R 3 is H, OH, or F; R 4 is selected from the group consisting of H, methyl, or OH; R 5 is selected from the group consisting of OH, CONH2, SO2NH2, or OSO2NH2; A compound of formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof:

[0080] In some embodiments, the compound is a compound of formula (1a): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; U is CH or N; V is CH or N; W is CH, COH, N, or N + -O -selected from the group consisting of: Z is selected from the group consisting of CH, COH, or N; X is CH or N; Y is CH, CF, COH, or N; R 1 and R 2 are each independently selected from the group consisting of H or methyl, or R 1 and R 2 together with the carbon atom to which they are attached form cyclopropyl; R 3 is H, OH, or F; R 4 is selected from the group consisting of H, methyl, or OH; R 5 is selected from the group consisting of OH, CONH2, SO2NH2, or OSO2NH2; A compound of formula (1a) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof:

[0081] In some embodiments, the compound is a compound of Formula (2), Formula (2a), or Formula (2b), where U, V, and Z can be CH and W can be COH.

[0082] In some embodiments, A can be selected from the group consisting of: COH, tetrazole, 1,2,4-oxadiazol-5(2H)-one, 1,3,4-oxadiazol-2(3H)-one, CONHSOR 6 , CONHSO2Me, SO3H, 1,3,4-oxadiazole-2(3H)-thione, 1,2,4-oxadiazole-5(2H)-thione, 1,2,4-thiadiazol-5(2H)-one, 1,2,5-thiadiazolidin-3-one 1,1-dioxide, and 2,4-oxazolidinedione.

[0083] In one embodiment, A may be selected from: [ka]

[0084] In some embodiments, A can be selected from CO2H and a tetrazole ring. In other embodiments, A can be CO2H. In other further embodiments, A can be a tetrazole ring.

[0085] In some embodiments, U can be CH. In other embodiments, U can be COH. In other embodiments, U can be N. In other embodiments, U can be N + -O - It could be.

[0086] In some embodiments, U is CH or N.

[0087] In some embodiments, V can be CH. In other embodiments, V can be COH. In other embodiments, V can be N. In other embodiments, V can be N + -O - It could be.

[0088] In some embodiments, V is CH or N.

[0089] In some embodiments, W can be CH. In other embodiments, W can be COH. In other embodiments, W can be N. In other embodiments, W can be N + -O - It could be.

[0090] In some embodiments, W is CH, COH, N, or N + -O - is selected from the group consisting of:

[0091] In some embodiments, Z can be CH. In other embodiments, Z can be COH. In other embodiments, Z can be N. In other embodiments, Z can be N + -O - It could be.

[0092] In some embodiments, Z is selected from the group consisting of CH, COH, or N.

[0093] In some embodiments, at least three of U, V, W, and Z are CH. In other embodiments, U, V, and W can be CH. In other embodiments, U, V, and Z can be CH. In other embodiments, U, W, and Z can be CH. In other embodiments, V, W, and Z can be CH. In other embodiments, U, V, W, and Z can be CH. In other embodiments, U, V, and Z can be CH. In other embodiments, U, V, and Z can be CH and W can be COH.

[0094] In some embodiments, X can be CH. In other embodiments, X can be CF. In other embodiments, X can be COH. In other embodiments, X can be N.

[0095] In some embodiments, X can be CH or N.

[0096] In some embodiments, Y can be CH. In other embodiments, Y can be CF. In other embodiments, Y can be COH. In other embodiments, Y can be N.

[0097] In some embodiments, Y can be CH, CF, COH, or N.

[0098] In some embodiments, at least one of X and Y is CH. In other embodiments, both X and Y can be CH.

[0099] In some embodiments, X can be CH or N and Y can be CH, CF, COH, or N.

[0100] In some embodiments, R 1 is H or C optionally substituted with 1 to 3 fluorine atoms1-3 In another embodiment, R 1 can be H. In another embodiment, R 1 is C optionally substituted with 1 to 3 fluorine atoms 1-3 In another embodiment, R 1 is C 1-3 In another embodiment, R 1 is R 2 C optionally substituted with 1 to 3 fluorine atoms linked to 3-6 In another embodiment, R 1 is R 2 Connected to C 3-6 In another embodiment, R 1 can be H or methyl, or R 2 may be linked to form a cyclopropane ring. 1 can be methyl optionally substituted with 1 to 3 fluorine atoms. In another embodiment, R 1 can be methyl. In another embodiment, R 1 is R 2 may be linked to form a cyclopropane ring optionally substituted with 1 to 3 fluorine atoms. 1 is R 2 may be linked to form a cyclopropane ring.

[0101] In some embodiments, R 2 can be H. In another embodiment, R 2 is C optionally substituted with 1 to 3 fluorine atoms 1-3 In another embodiment, R 2 is C 1-3 In another embodiment, R 2 is R 1 C optionally substituted with 1 to 3 fluorine atoms linked to 3-6 In another embodiment, R 2 is R1 Connected to C 3-6 In another embodiment, R 2 is R 1 may be linked to form a cyclopropane ring optionally substituted with 1 to 3 fluorine atoms. 2 is R 1 may be linked to form a cyclopropane ring.

[0102] In some embodiments, R 3 can be H. In another embodiment, R 3 can be OH. In another embodiment, R 3 can be F.

[0103] In some embodiments, R 3 can be H, OH, or F.

[0104] In some embodiments, R 3 can be H or F.

[0105] In some embodiments, R 4 and R 5 together with the ring to which they are attached may be selected from the group consisting of: [ka]

[0106] In some embodiments, R 4 can be H. R 4 can be OH. In another embodiment, R 4 can be CN. In another embodiment, R 4 can be halo. In another embodiment, R 4 can be F. In another embodiment, R 4 can be Cl. In another embodiment, R 4 can be Br. In another embodiment, R 4 is C optionally substituted with 1 to 3 fluorine atoms1-3 In another embodiment, R 4 is C 1-3 In another embodiment, R 4 can be methoxy optionally substituted with 1 to 3 fluorine atoms. 4 In another embodiment, R 4 is C optionally substituted with 1 to 3 fluorine atoms 1-3 In another embodiment, R 4 is C 1-3 In another embodiment, R 4 can be H or methyl. In another embodiment, R 4 can be methyl optionally substituted with 1 to 3 fluorine atoms. In another embodiment, R 4 can be methyl.

[0107] In some embodiments, R 4 is H, OH, CN, halo, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkoxy or C optionally substituted with 1 to 3 fluorine atoms 1-3 alkyl.

[0108] In some embodiments, R 5 can be OH. In another embodiment, R 5 can be COH. In another embodiment, R 5 can be CONH. In another embodiment, R 5 can be SO2NH2. In another embodiment, R 5 can be OSO2NH2.

[0109] In some embodiments, R 5 can be OH, CONH2, SO2NH2, or OSO2NH2.

[0110] In some embodiments, R 6 is C1-3 In another embodiment, R 6 is C 3-6 In another embodiment, R 6 can be methyl.

[0111] In some embodiments, the compounds or pharmaceutically acceptable salts disclosed herein have EP4 receptor agonist activity.

[0112] In some embodiments, disclosed herein are methods for treating an EP4 receptor-mediated disease, comprising administering to a patient in need of treatment for an EP4 receptor-mediated disease a compound or pharmaceutically acceptable salt disclosed herein.

[0113] In some embodiments, the method of treating an EP4 receptor-mediated disease is a gastrointestinal disorder.

[0114] In some embodiments, the gastrointestinal disorder is selected from the group consisting of constipation, 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 dysfunction, postoperative ileus, food allergy or intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced bowel disease, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrhea, immune-mediated gastrointestinal disorders, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis, or pulmonary diseases and conditions such as chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, reactive airways disease, and idiopathic pulmonary fibrosis.

[0115] 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.

[0116] In some embodiments, the compound of Formula I or Formula (1) is a compound listed in Table 1, or a pharmaceutically acceptable salt thereof. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] Abbreviation aq aqueous solution Bn Benzyl DCM dichloromethane DMA Dimethylacetamide DMF Dimethylformamide dppf 1,1'-bis(diphenylphosphino)ferrocene EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EtOAc ethyl acetate HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate HOBt Hydroxybenzotriazole HPLC High-Performance Liquid Chromatography hr time hrs time LC / MS Liquid Chromatography Mass Spectrometry M molar concentration MeCN acetonitrile MeOH Methanol N specified concentration Prep HPLC Preparative High Performance Liquid Chromatography RT room temperature sat saturation THF tetrahydrofuran UPLC Ultra High Performance Liquid Chromatography

[0117] definition For the purposes of this application, the following definitions apply unless otherwise indicated.

[0118] The term "treatment," in connection with the use of any of the compounds described herein, including compounds of Formula I, Formula (1), Formula (1a), Formula (1b), Formula (2), Formula (2a), Formula (2b), Formula (3), Formula (3a), Formula (3b), Formula (4), Formula (4a), and Formula (4b), is used to describe any form of intervention in which a compound is administered to a subject suffering from, at risk of suffering from, or potentially at risk of suffering from the disease or disorder in question. That is, the term "treatment" encompasses both prophylactic treatment and treatment when measurable or detectable symptoms of the disease or condition are present.

[0119] The term "therapeutically effective amount" (e.g., in connection with a method of treating a disease or condition) refers to an amount of a compound effective to produce a desired therapeutic effect. For example, if the condition is pain, a therapeutically effective amount is an amount sufficient to provide a desired level of analgesia. The desired level of analgesia can be, for example, complete elimination of pain or a reduction in the severity of pain.

[0120] As used herein, the term "hydroxyl" or "hydroxy" refers to an --OH moiety.

[0121] As used herein, an "alkyl" group 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-chained or branched. 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. Alkyl groups include halo, phospho, cycloaliphatic [e.g., cycloalkyl or cycloalkenyl], heterocycloaliphatic [e.g., heterocycloalkyl or heterocycloalkenyl], aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl [e.g., (aliphatic)carbonyl, (cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonyl], nitro, cyano, amido [e.g., (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino], and the like. It may be substituted (i.e., optionally substituted) with one or more substituents such as alkylaminocarbonyl, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, arylaminocarbonyl, or heteroarylaminocarbonyl], amino [e.g., aliphatic amino, cycloaliphatic amino, or heterocycloaliphatic amino], sulfonyl [e.g., aliphatic -SO2-], sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamido, oxo, carboxy, carbamoyl, cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, or hydroxy.Some examples of substituted alkyl include, but are not limited to, carboxyalkyl (such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, aralkyl, (alkoxyaryl)alkyl, (sulfonylamino)alkyl (such as (alkyl-SO2-amino)alkyl), aminoalkyl, amidoalkyl, (cycloaliphatic)alkyl, or haloalkyl.

[0122] As used herein, an "alkylene" group or a "cycloalkane" group refers to a divalent branched, straight-chain, or cyclic alkyl group containing 2 to 12 (e.g., 2 to 8, 2 to 6, or 2 to 4) carbon atoms and serving to connect two chemical moieties. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, butylene, isopropylene (methylethylene), and isobutylene (2-methylpropylene). An alkylene group or a cycloalkane group can be substituted (i.e., optionally substituted) with one or more substituents as defined for an alkyl group.

[0123] As used herein, "amide" encompasses both "aminocarbonyl" and "carbonylamino." These terms, when used alone or in conjunction with another group, include -N(R X )-C(O)-R Y or -C(O)-N(R X )2, and -C(O)-N(R when used internally X )- or -N(R X )-C(O)-, and R X and R Ycan be aliphatic, cycloaliphatic, aryl, araliphatic, heterocycloaliphatic, heteroaryl, or heteroaraliphatic. Examples of amido groups include alkylamido (such as alkylcarbonylamino or alkylaminocarbonyl), (heterocycloaliphatic)amido, (heteroaralkyl)amido, (heteroaryl)amido, (heterocycloalkyl)alkylamido, arylamido, aralkylamido, (cycloalkyl)alkylamido, or cycloalkylamido.

[0124] As used herein, an "amino" group is -NR X R Y In the formula, R X and R Y and each independently represent hydrogen, aliphatic, cycloaliphatic, (cycloaliphatic)aliphatic, aryl, araliphatic, heterocycloaliphatic, (heterocycloaliphatic)aliphatic, heteroaryl, carboxy, sulfanyl, sulfinyl, sulfonyl, (aliphatic)carbonyl, (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphatic)carbonyl, arylcarbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl, ((heterocycloaliphatic)aliphatic)carbonyl, (heteroaryl)carbonyl, or (heteroaraliphatic)carbonyl, each of which is defined herein and optionally substituted. Examples of amino groups include alkylamino, dialkylamino, or arylamino. The term "amino" when not a terminal group (e.g., alkylcarbonylamino) does not include -NR X -, where R X has the same meaning as defined above.

[0125] As used herein, the term "aryl," used alone or as part of a larger moiety, as in "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to monocyclic (e.g., phenyl); bicyclic (e.g., indenyl, naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl); and tricyclic (e.g., fluorenyl, tetrahydrofluorenyl, or tetrahydroanthracenyl, anthracenyl) ring systems, where the monocyclic ring system is aromatic or at least one ring in a bicyclic or tricyclic ring system is aromatic. Bicyclic and tricyclic groups encompass benzo-fused 2- to 3-membered carbocyclic rings. For example, benzo-fused groups include groups having two or more C 4-8 and phenyl fused to a carbocyclic moiety. Aryl is optionally aliphatic (e.g., alkyl, alkenyl, or alkynyl); cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; oxo (on the non-aromatic carbocyclic ring of a benzo-fused bicyclic or tricyclic aryl); nitro; carboxy; amido; acyl (e.g., (aliphatic)carbonyl; (cycloaliphatic)oxy; The aryl may be substituted with one or more substituents, including (aliphatic)carbonyl; ((cycloaliphatic)aliphatic)carbonyl; (araliphatic)carbonyl; (heterocycloaliphatic)carbonyl; ((heterocycloaliphatic)aliphatic)carbonyl; or (heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphatic -SO2- or amino-SO2-]; sulfinyl [e.g., aliphatic -S(O)- or cycloaliphatic -S(O)-]; sulfanyl [e.g., aliphatic -S-]; cyano; halo; hydroxy; mercapto; sulfoxy; urea; thiourea; sulfamoyl; sulfamido; or carbamoyl. Alternatively, the aryl may be unsubstituted.

[0126] Non-limiting examples of substituted aryls include haloaryls [e.g., mono-, di-(p,m-dihaloaryls, etc.), and (trihalo)aryls]; (carboxy)aryls [e.g., (alkoxycarbonyl)aryls, ((aralkyl)carbonyloxy)aryls, and (alkoxycarbonyl)aryls]; (amido)aryls [e.g., (aminocarbonyl)aryls, (((alkylamino)alkyl)aminocarbonyl)aryls, (alkylcarbonyl)aminoaryls, (arylaminocarbonyl)aryls, and (((heteroaryl)amino)carbonyl)aryls]; aminoaryls [e.g., ((alkylsulfonyl)amino)aryls, or ((dialkyl)amino)aryls]; (cyanoalkyl)aryls; (alkoxy)aryls; (sulfamoyl)aryls [e.g., (aminosulfonyl)amino (m-(heterocycloaliphatic)-o-(alkyl)aryl; (((m-(m-alkoxy)aryl); (hydroxy)aryl, ((carboxy)alkyl)aryl; (((dialkyl)amino)alkyl)aryl; (nitroalkyl)aryl; (((alkylsulfonyl)amino)alkyl)aryl; ((heterocycloaliphatic)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-(heterocycloaliphatic)-o-(alkyl))aryl.

[0127] 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. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, norbornyl, cubyl, 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.

[0128] Cycloalkyl groups are optionally selected from the group consisting of phospho, aliphatic [e.g., alkyl, alkenyl, or alkynyl], cycloaliphatic, (cycloaliphatic)aliphatic, heterocycloaliphatic, (heterocycloaliphatic)aliphatic, aryl, heteroaryl, alkoxy, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy, aryloxy, heteroaryloxy, (araliphatic)oxy, (heteroaraliphatic)oxy, aroyl, heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino, (cycloaliphatic)carbonylamino, ((cycloaliphatic)aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino, (heterocycloaliphatic)carbonylamino, ((heterocycloaliphatic)aliphatic)carbonylamino, (heteroaryl)carbonylamino, and (heteroaraliphatic)carbonylamino, or (heteroaraliphatic)carbonylamino], nitro, carboxy [e.g., HOOC-, alkoxycarbonyl, or alkylcarbonyloxy], acyl [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl, ((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaraliphatic)carbonyl], cyano, halo, hydroxy, mercapto, sulfonyl [e.g., alkyl-SO2- and aryl-SO2-], sulfinyl [e.g., alkyl-S(O)-], sulfanyl [e.g., alkyl-S-], sulfoxy, urea, thiourea, sulfamoyl, sulfamido, oxo, or carbamoyl.

[0129] As used herein, a "heterocycloalkyl" group refers to a 3- to 10-membered monocyclic or bicyclic (fused or bridged) (e.g., 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 combinations thereof). Examples of heterocycloalkyl groups include piperidyl, piperazyl, tetrahydropyranyl, tetrahydrofuryl, 1,4-dioxolanyl, 1,4-dithianyl, 1,3-dioxolanyl, oxazolidyl, isoxazolidyl, morpholinyl, thiomorpholyl, octahydrobenzofuryl, octahydrochromenyl, octahydrothiochromenyl, octahydroindolyl, octahydropyrindinyl, decahydroquinolinyl, octahydrobenzo[b]thiophenyl, 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]octyl. 3,7 Monocyclic heterocycloalkyl groups can be fused with a phenyl moiety to form structures such as tetrahydroisoquinoline, which would be classified as heteroaryl.

[0130] Heterocycloalkyl groups are optionally selected from the group consisting of phospho, aliphatic [e.g., alkyl, alkenyl, or alkynyl], cycloaliphatic, (cycloaliphatic)aliphatic, heterocycloaliphatic, (heterocycloaliphatic)aliphatic, aryl, heteroaryl, alkoxy, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy, aryloxy, heteroaryloxy, (araliphatic)oxy, (heteroaraliphatic)oxy, aroyl, heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino, (cycloaliphatic)carbonylamino, ((cycloaliphatic)aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino, (heterocycloaliphatic)carbonylamino, ((heterocycloaliphatic)aliphatic)carbonylamino, (heteroaryl)carbonyl], and the like. and (heteroaraliphatic)amino, or (heteroaraliphatic)carbonylamino], nitro, carboxy [e.g., HOOC-, alkoxycarbonyl, or alkylcarbonyloxy], acyl [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl, ((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaraliphatic)carbonyl], nitro, cyano, halo, hydroxy, mercapto, sulfonyl [e.g., alkylsulfonyl or arylsulfonyl], sulfinyl [e.g., alkylsulfinyl], sulfanyl [e.g., alkylsulfanyl], sulfoxy, urea, thiourea, sulfamoyl, sulfamido, oxo, or carbamoyl.

[0131] A "heteroaryl" group, as used herein, 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 ring system is aromatic, or at least one of the rings in the bicyclic or tricyclic ring system is aromatic.

[0132] Heteroaryl groups include benzo-fused ring systems having 2 to 3 rings. For example, a benzo-fused group includes benzo fused to one or two 4- to 8-membered heterocycloaliphatic moieties (e.g., indolyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furyl, benzo[b]thiophen-yl, quinolinyl, or isoquinolinyl). Some examples of heteroaryl are azetidinyl, pyridyl, 1H-indazolyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuryl, isoquinolinyl, benzthiazolyl, xanthene, thioxanthene, phenothiazine, dihydroindole, benzo[1,3]dioxole, benzo[b]furyl, benzo[b]thiophenyl, indazolyl, benzimidazolyl, benzthiazolyl, puryl, cinnolyl, quinolyl, quinazolyl, cinnolyl, phthalazyl, quinazolyl, quinoxalyl, isoquinolyl, 4H-quinolidyl, benzo-1,2,5-thiadiazolyl, or 1,8-naphthyridyl.

[0133] Heteroaryl groups include N-oxide heteroaryl compounds, such as pyridine, pyrimidine, pyrazine, pyrrole, imidazole, thiazole, quinoline, or isoquinoline, in which oxidation occurs at the nitrogen atom. Examples of N-oxide heteroaryls include those of the formula C5H5N - →O + There are pyridine-N-oxides having the formula:

[0134] Monocyclic heteroaryls include, but are not limited to, furyl, thiophen-yl, 2H-pyrrolyl, pyrrolyl, oxazolyl, thazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4H-pranyl, pyridyl, pyridazyl, pyrimidyl, pyrazolyl, pyrazyl, or 1,3,5-triazyl. Monocyclic heteroaryls are numbered according to standard compound nomenclature.

[0135] Bicyclic heteroaryls include, but are not limited to, indolyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furyl, benzo[b]thiophenyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, indolyl, benzo[b]furyl, benzo[b]thiophenyl, indazolyl, benzimidazyl, benzthiazolyl, purinyl, 4H-quinolizyl, quinolyl, isoquinolyl, cinnolyl, phthalazyl, quinazolyl, quinoxalyl, 1,8-naphthyridyl, or pteridyl. Bicyclic heteroaryls are numbered according to standard compound nomenclature.

[0136] Heteroaryl is optionally aliphatic [e.g., alkyl, alkenyl, or alkynyl]; cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; oxo (on a non-aromatic carbocyclic or heterocyclic ring of a bicyclic or tricyclic heteroaryl); carboxy; amido; acyl [e.g., aliphatic carbonyl; Heteroaryl may be substituted with one or more substituents such as (cycloaliphatic)carbonyl; ((cycloaliphatic)aliphatic)carbonyl; (araliphatic)carbonyl; (heterocycloaliphatic)carbonyl; ((heterocycloaliphatic)aliphatic)carbonyl; or (heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphatic sulfonyl or aminosulfonyl]; sulfinyl [e.g., aliphatic sulfinyl]; sulfanyl [e.g., aliphatic sulfanyl]; nitro; cyano; halo; hydroxy; mercapto; sulfoxy; urea; thiourea; sulfamoyl; sulfamido; or carbamoyl. Alternatively, the heteroaryl may be unsubstituted.

[0137] Non-limiting examples of substituted heteroaryls include (halo)heteroaryls [e.g., mono- and di-(halo)heteroaryls]; (carboxy)heteroaryls [e.g., (alkoxycarbonyl)heteroaryls]; cyanoheteroaryls; aminoheteroaryls [e.g., ((alkylsulfonyl)amino)heteroaryls and ((dialkyl)amino)heteroaryls]; (amido)heteroaryls [e.g., aminocarbonylheteroaryls, ((alkylcarbonyl)amino)heteroaryls, ((((alkyl)amino)alkyl)aminocarbonyl)heteroaryls, (((heteroaryl)amino)carbonyl)heteroaryls, ((heterocycloaliphatic)carbonyl)heteroaryls, and ((alkylcarbonyl)amino)heteroaryls]; (cyanoalkyl)heteroaryls; (alkoxy)heteroaryls; (sulfamoyl)heteroaryls. (aminosulfonyl)heteroaryl; (sulfonyl)heteroaryl [e.g., (alkylsulfonyl)heteroaryl]; (hydroxyalkyl)heteroaryl; (alkoxyalkyl)heteroaryl; (hydroxy)heteroaryl; ((carboxy)alkyl)heteroaryl; (((dialkyl)amino)alkyl]heteroaryl; (heterocycloaliphatic)heteroaryl; (cycloaliphatic)heteroaryl; (nitroalkyl)heteroaryl; (((alkylsulfonyl)amino)alkyl)heteroaryl; ((alkylsulfonyl)alkyl)heteroaryl; (cyanoalkyl)heteroaryl; (acyl)heteroaryl [e.g., (alkylcarbonyl)heteroaryl]; (alkyl)heteroaryl; or (haloalkyl)heteroaryl [e.g., trihaloalkylheteroaryl].

[0138] As used herein, an "alkoxy" group refers to an alkyl-O- group, where "alkyl" is defined above.

[0139] As used herein, a "carboxy" group refers to a -COOH, -COOR group when used as a terminal group. X , -OC(O)H, -OC(O)R Xor, when used as an internal group, refers to -OC(O)- or -C(O)O-.

[0140] As used herein, a "mercapto" group refers to -SH.

[0141] As used herein, a "sulfo" group refers to -SO3H or -SO3R when used terminally. X , or -S(O)3- when used internally.

[0142] As used herein, a "sulfamido" group refers to a group having the structure -NR when used terminally. X -S(O)2-NR Y R Z , and -NR when used internally X -S(O)2-NR Y - refers to R X , R Y , and R Z is defined above.

[0143] As used herein, a "sulfamoyl" group refers to a group having the structure -OS(O)2-NR Y R Z refers to R Y and R Z is defined above.

[0144] As used herein, a "sulfonamide" group refers to a group having the structure -S(O)-NR when used terminally. X R y or -NR x -S(O)2-R z or -S(O)2-NR when used internally x -or-NR x -S(O)2-, R x , R y , and R Z is defined above.

[0145] As used herein, a "sulfanyl" group refers to a group such as -SR when used terminally. X , and when used internally refers to -S-, R X is defined above. Examples of sulfanyl include aliphatic-S-, cycloaliphatic-S-, aryl-S-, or the like.

[0146] As used herein, a "halogen" or "halo" group refers to fluorine, chlorine, bromine, or iodine.

[0147] As used herein, "oxo" refers to =O.

[0148] As used herein, the term "vicinal" generally refers to the positioning of substituents on a group that includes two or more carbon atoms, the substituents being attached to adjacent carbon atoms.

[0149] As used herein, the term "geminal" generally refers to the placement on a group that includes two or more carbon atoms of substituents that are attached to the same carbon atom.

[0150] The terms "terminally" and "internally" refer to the location of a group within a substituent. A group is terminal if it is at the end of the substituent and is not further attached to the rest of the chemical structure. Carboxyalkyl, i.e., R X O(O)C-alkyl is an example of a carboxy group used terminally. When a group is present in the middle of a substituent in a chemical structure, the group is internal. Alkylcarboxy (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 examples of carboxy groups used internally.

[0151] As used herein, "aliphatic chain" refers to a branched or straight-chain aliphatic group (e.g., an alkyl group, an alkenyl group, or an alkynyl group). A straight-chain aliphatic chain is a group having a structure similar to -[CH2]v -structure, and v is 1 to 12. A branched aliphatic chain is a linear aliphatic chain substituted with one or more aliphatic groups. A branched aliphatic chain has a -[CQQ] v - structure, and each Q is, independently, hydrogen or an aliphatic group, provided that at least one Q is an aliphatic group. The term aliphatic chain encompasses alkyl chains, alkenyl chains, and alkynyl chains, where alkyl, alkenyl, and alkynyl are defined above.

[0152] The phrase "optionally substituted" is used interchangeably herein with the phrase "substituted or unsubstituted." As described herein, the compounds of the present invention may be optionally substituted with one or more substituents, as exemplified by the general formulas above or by the specific classes, subclasses, and species of the present invention. Unless otherwise specified, each of the specific groups of variables described herein may be optionally substituted with one or more substituents as described herein. Each substituent of a specific group may be optionally further substituted with one to three of halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, cycloaliphatic, heterocycloaliphatic, heteroaryl, haloalkyl, and alkyl. For example, an alkyl group may be substituted with alkylsulfanyl, which may be optionally substituted with one to three of halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, haloalkyl, and alkyl. As an additional example, the cycloalkyl portion of a (cycloalkyl)carbonylamino can be optionally substituted with one to three of halo, 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 be combined with the atoms to which they are bound to form a ring.

[0153] As used herein, the term "substituted," whether preceded by the term "optionally," generally refers to the replacement of a hydrogen atom in a given structure with the radical of a specified substituent. Specific substituents are described above in the definitions and in the description of compounds and examples below. Unless otherwise specified, an 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 with more than one substituent selected from a specified group, those substituents may be the same or different at each position. A ring substituent, such as a heterocycloalkyl, can be attached to another ring, such as a cycloalkyl, to form a spiro-bicyclic ring system (e.g., both rings share one common atom). As one of skill in the art will recognize, combinations of substituents envisioned by this invention are those that result in the formation of stable or chemically feasible compounds.

[0154] As used herein, the phrase "stable or chemically feasible" refers to a compound that does not change substantially when subjected to conditions that allow for its production, detection, and preferably 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 does not change substantially when kept at a temperature of 40° C. or below, in the absence of moisture, or other chemically reactive conditions, for at least one week.

[0155] Where any of the compounds described contain chiral centers, the invention extends to all optical isomers of such compounds, whether in the form of racemates or resolved enantiomers. The invention described herein relates to all crystalline forms, solvates, and hydrates of any of the disclosed compounds, however prepared.

[0156] When any of the compounds disclosed herein contain an acidic or basic center, such as a carboxylic acid or amino group, all salt forms of said compounds are included in the present invention. The invention described herein further relates to all tautomers of the compounds presented herein. For pharmaceutical purposes, the salts should be considered to be pharmaceutically acceptable salts.

[0157] The salts or pharmaceutically acceptable salts that may be mentioned include acid addition salts and base addition salts.Such salts can be formed by conventional means, for example, by reacting the free acid or free base form of the compound with one or more equivalents of a suitable acid or base, optionally in a solvent or in a medium in which the salt is insoluble, followed by removing the solvent or medium using standard techniques (for example, in vacuo, lyophilization, or filtration).Salts can also be prepared by exchanging the counterion of the compound in the form of a salt with another counterion, for example, using a suitable ion exchange resin.

[0158] Examples of pharmaceutically acceptable salts include acid addition salts derived from mineral and organic acids, as well as salts derived from metals such as sodium, magnesium, potassium, and calcium.

[0159] Examples of acid addition salts include 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, butanoic acid, (+) camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic 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., 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- Included are acid addition salts formed with 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-amino-salicylic 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.

[0160] Also included are any solvates of the compounds and their salts. Preferred solvates are those formed by incorporating molecules of a non-toxic, pharmaceutically acceptable solvent (hereinafter referred to as the solvating solvent) into the solid structure (e.g., crystalline structure) of the compounds of the present invention. Examples of such solvents include water, alcohols (such as ethanol, isopropanol, and butanol), and dimethyl sulfoxide. Solvates can be prepared by recrystallizing the compounds of the present invention with a solvent or a mixture of solvents containing the solvating solvent. Whether a solvate has formed in any given case can be determined by analyzing the crystals of the compound using well-known standard techniques, such as thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and X-ray crystallography.

[0161] Solvate can be stoichiometric or non-stoichiometric solvate.Particular solvate can be hydrate, and examples of hydrate include hemihydrate, monohydrate and dihydrate.For more detailed discussion of solvate and the methods used to prepare and characterize solvate, see Bryn et al., Solid-State Chemistry of Drugs, Second Edition, Publisher SSCI, Inc. (West Lafayette, Indiana, USA), 1999, ISBN0-967-06710-3.

[0162] In the context of the present invention, the term "pharmaceutical composition" refers to a composition containing an active agent and further containing one or more pharmaceutically acceptable carriers or excipients. Depending on the mode of administration and the nature of the dosage form, the composition may further contain components selected from, for example, diluents, adjuvants, excipients, vehicles, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavorings, fragrances, antibacterial agents, antifungal agents, lubricants, and dispersing agents. The composition may take the form of, for example, tablets, dragees, powders, elixirs, syrups, liquid preparations including suspensions, sprays, inhalants, tablets, confectionery tablets, emulsions, solutions, cachets, granules, capsules, and suppositories, as well as injectable liquid preparations including liposomal preparations.

[0163] The compounds of the invention may contain one or more isotopic substitutions, and a reference to a particular element includes within its scope all isotopes of that element. For example, a reference to hydrogen includes within its scope: 1 H, 2 H(D), and 3 Similarly, references to carbon and oxygen include within their scope: 12 C. 13 C, and 14 C and 16 O and 18 and O, respectively. Similarly, reference to a particular functional group also includes within its scope isotopic variations unless the context indicates otherwise. For example, reference to an alkyl group such as an ethyl group or an alkoxy group such as a methoxy group also includes variations in which one or more of the group's hydrogen atoms are in the form of a deuterium or tritium isotope, such as, for example, an ethyl group in which all five hydrogen atoms are in the deuterium isotope form (a perdeuteroethyl group), or a methoxy group in which all three hydrogen atoms are in the deuterium isotope form (a trideuteromethoxy group). Isotopes may be radioactive or non-radioactive.

[0164] Therapeutic dosage can vary depending on the patient's requirements, the severity of the condition being treated, and the compound being used.Determining the dosage that is appropriate for a particular situation is within the skill of a person skilled in the art.Generally, treatment is initiated with a smaller dosage that is less than the appropriate amount of the compound.Then, the dosage is gradually increased until the optimal effect under the situation is reached.For convenience, optionally, the total daily dosage can be divided and administered in portions throughout the day.

[0165] The magnitude of an effective amount of a compound will, of course, vary depending on the nature of the severity of the condition being treated, as well as the particular compound and its route of administration. Selecting an appropriate dose is within the ability of one skilled in the art and does not pose an undue burden. Generally, the daily dose range can be about 10 μg to about 30 mg per kg of body weight for humans and non-human animals, preferably about 50 μg to about 30 mg per kg of body weight for humans and non-human animals, for example, about 50 μg to about 10 mg per kg of body weight for humans and non-human animals, for example, about 100 μg to about 30 mg per kg of body weight for humans and non-human animals, for example, about 100 μg to about 10 mg per kg of body weight for humans and non-human animals, and most preferably about 100 μg to about 1 mg per kg of body weight for humans and non-human animals.

[0166] Combination therapy An effective amount can be achieved in a method or pharmaceutical composition of the invention using a compound of the invention (including a pharmaceutically acceptable salt or solvate (e.g., hydrate)) alone or in combination with an additional appropriate therapeutic agent, such as an antiviral agent or a vaccine. When "combination therapy" is used, an effective amount can be achieved using a first amount of a compound of the invention and a second amount of an additional appropriate therapeutic agent.

[0167] In another embodiment of the invention, the compound of the invention and the additional therapeutic agent are each administered in an effective amount (i.e., each in an amount that would be therapeutically effective if administered alone). In another embodiment, the compound of the invention and the additional therapeutic agent are each administered in an amount that would not provide a therapeutic effect alone (a subtherapeutic dose). In yet another embodiment, the compound of the invention can be administered in an effective amount while the additional therapeutic agent is administered in a subtherapeutic dose. In yet another embodiment, the compound of the invention can be administered in a subtherapeutic dose while the additional therapeutic agent, e.g., a suitable cancer therapeutic agent, is administered in an effective amount.

[0168] As used herein, the terms "in combination" or "co-administration" can be used interchangeably to refer to the use of two or more therapies (e.g., one or more prophylactic and / or therapeutic agents). The use of these terms does not restrict the order in which therapies (e.g., prophylactic and / or therapeutic agents) are administered to a subject.

[0169] Simultaneous administration includes essentially simultaneous administration of a first and second amount of the compound, for example, in the form of a single pharmaceutical composition, such as a capsule or tablet, having a fixed ratio of the first and second amounts, or in the form of multiple separate capsules or tablets for each. In addition, such simultaneous administration also includes use of each compound sequentially, in any order.

[0170] In one embodiment, a compound of the invention and an additional therapeutic agent are administered separately, sequentially, or simultaneously to a subject.

[0171] When simultaneous administration involves separate administration of a first amount of a compound of the present invention and a second amount of an additional therapeutic agent, these compounds are administered close enough in time to produce the desired therapeutic effect. For example, the period between each administration that can produce the desired therapeutic effect can range from several minutes to several hours and can be determined by taking into account the properties of each compound, such as potency, solubility, bioavailability, plasma half-life, and kinetic profile. For example, the compound of the present 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.

[0172] 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), concomitantly with, or subsequent 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 after) the administration of a second therapy (e.g., a prophylactic or therapeutic agent such as an anti-cancer agent).

[0173] It is understood that the simultaneous administration of a first amount of a compound of the invention and a second amount of an additional therapeutic agent can result in an enhanced or synergistic therapeutic effect, which combined effect is greater than the additive effect that would result from separate administration of a first amount of a compound of the invention and a second amount of an additional therapeutic agent.

[0174] As used herein, the term "synergistic" refers to a combination of a compound of the present invention and another therapy (e.g., a prophylactic or therapeutic agent) that is more effective than the additive effects of the therapies. A synergistic effect of a combination of therapies (e.g., a combination of prophylactic or therapeutic agents) may allow a subject to use lower dosages of one or more of the therapies and / or less frequent administration of the therapies. The availability of lower dosages of a therapy (e.g., a prophylactic or therapeutic agent) and / or the ability to administer the therapy less frequently may reduce the toxicity associated with administering the therapy to a subject without reducing the efficacy of the therapy in preventing, managing, or treating a disorder. In addition, a synergistic effect may improve the efficacy of each agent in preventing, managing, or treating a disorder. Finally, a synergistic effect of a combination of therapies (e.g., a combination of prophylactic or therapeutic agents) may avoid or reduce adverse or unwanted side effects associated with the use of either therapy alone.

[0175] The presence or absence of synergy can be determined using a suitable method for assessing drug interactions. Suitable methods include, for example, the sigmoid-E equation (Holford, NHG and Scheiner, LB, Clin. Pharmacokinet. 6: 429-453 (1981)), the Loewe additivity equation (Loewe, S. and Muischnek, H., Arch. Exp. Pathol Pharmacol. 114: 313-326 (1926)), and the median-effect equation (Chou, TC and Talalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). By applying these equations to experimental data, corresponding graphs can be generated to aid in assessing the effectiveness of drug combinations. The corresponding graphs associated with the above equations are the concentration-effect curve, the isobologram curve, and the combination index curve, respectively.

[0176] 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 formulated together. 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 formulated separately.

[0177] In another aspect, the invention provides a kit comprising a compound of the invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and at least one additional therapeutic agent. The kit may include instructions for administering the compound of the invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and at least one additional therapeutic agent to a subject in need thereof.

[0178] In another aspect, the present invention provides a combination therapy for use as a pharmaceutical, comprising administering a compound of the present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer to a subject in need thereof, and administering an additional therapeutic agent to a subject in need thereof.

[0179] The compound of the invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and the additional therapeutic agent may be administered to a subject separately, sequentially, or simultaneously.

[0180] The pharmaceutical composition, kit, and / or combination therapy may be for use in treating a gastrointestinal disorder or a pulmonary disease or condition. The gastrointestinal disorder may be selected from the group consisting of constipation, 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 dysfunction, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorder, functional gastrointestinal disorder, drug-induced bowel disease, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrhea, immune-mediated gastrointestinal disease, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis. The pulmonary disease or condition may be selected from the group consisting of chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, reactive airway disease, and idiopathic pulmonary fibrosis.

[0181] The at least one additional therapeutic agent may be selected from the group consisting of an aminosalicylate, a corticosteroid, an immunomodulator, and combinations thereof. An aminosalicylates is also known as 5-aminosalicylate (5-ASA). The at least one additional therapeutic agent may be an aminosalicylate. The aminosalicylate may be mesalamine. The aminosalicylate may be sulfasalazine. The at least one additional therapeutic agent may be a corticosteroid. The corticosteroid may be budesonide. The at least one additional therapeutic agent may be an immunomodulator. The immunomodulator may be a thiopurine. The immunomodulator may be methotrexate.

[0182] Pharmaceutical Composition While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (eg, a formulation).

[0183] Thus, in another embodiment of the present invention, there is provided a pharmaceutical composition comprising at least one compound of formula I as defined above, together with at least one pharmaceutically acceptable excipient.

[0184] When the pharmaceutical composition includes at least one additional therapeutic agent, the compound of the invention and the at least one additional therapeutic agent can be formulated together, or the compound of the invention and the at least one additional therapeutic agent can be formulated separately.

[0185] The composition may be a tablet composition.

[0186] The composition may be a capsule composition.

[0187] Pharmaceutically acceptable excipients may be selected from, for example, carriers (e.g., solid, liquid, or semi-solid carriers), adjuvants, diluents (e.g., solid diluents such as fillers or bulking agents; and liquid diluents such as solvents and co-solvents), granulating agents, binders, flow aids, coating agents, release-controlling agents (e.g., polymers or waxes that retard or delay release), binding agents, disintegrants, buffers, lubricants, preservatives, antifungal and antibacterial agents, antioxidants, buffers, isotonicity agents, thickeners, flavors, sweeteners, pigments, plasticizers, taste-masking agents, stabilizers, or any other excipient conventionally used in pharmaceutical compositions.

[0188] The term "pharmaceutically acceptable," as used herein, means that the compound, substance, composition, and / or dosage form is, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject (e.g., a human subject) without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable risk / benefit ratio. Each excipient must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

[0189] Pharmaceutical compositions containing compounds of formula I can be formulated according to known techniques, see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, USA.

[0190] The pharmaceutical composition can be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic, otic, rectal, vaginal, or transdermal administration.

[0191] Pharmaceutical dosage forms suitable for oral administration include tablets (coated or uncoated), capsules (hard or soft), caplets, pills, lozenges, syrups, solutions, powders, granules, elixirs and suspensions, sublingual tablets, wafers, or patches such as buccal patches.

[0192] The tablet composition may contain a unit dose of active compound together with an inert diluent or carrier, such as a sugar or sugar alcohol (e.g., lactose, sucrose, sorbitol, or mannitol); and / or a non-sugar derived diluent (sodium carbonate, calcium phosphate, calcium carbonate, or cellulose or its derivatives (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 (e.g., 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 buffer or citrate buffer), and effervescent agents (e.g., citrate / bicarbonate mixtures). Such excipients are well known and need not be discussed at length herein.

[0193] Tablets may be designed to release the drug upon contact with gastric fluids (immediate-release tablets) or may be designed to release in a controlled manner over an extended period of time or in specific areas of the gastrointestinal tract (controlled-release tablets).

[0194] Pharmaceutical compositions typically contain about 1% (w / w) to about 95% (w / w), preferably 1% (w / w), of the active ingredient and 99% (w / w) to 5% (w / w) of a pharmaceutically acceptable excipient (e.g., as defined above) or a combination of such excipients. Preferably, the composition contains about 20% (w / w) to about 90% (w / w) of the active ingredient and 80% (w / w) to 10% (w / w) of a pharmaceutically acceptable excipient or a combination of excipients. Pharmaceutical compositions contain about 1% to about 95%, preferably about 20% to about 90%, of the active ingredient. Pharmaceutical compositions of the present invention may be, for example, in unit dose form, such as in the form of an ampule, vial, suppository, prefilled syringe, dragee, powder, tablet, or capsule.

[0195] Tablets and capsules may contain, for example, 0-20% disintegrant, 0-5% lubricant, 0-5% flow aid, and / or 0-99% (w / w) filler or bulking agent (depending on drug dose). Tablets and capsules may also contain 0-10% (w / w) polymeric binder, 0-5% (w / w) antioxidant, and 0-5% (w / w) pigment. Sustained-release tablets will also typically contain 0-99% (w / w) release-controlling (e.g., retarding) polymer (depending on dose). Tablet or capsule film coatings typically contain 0-10% (w / w) polymer, 0-3% (w / w) pigment, and / or 0-2% (w / w) plasticizer.

[0196] Parenteral formulations typically contain 0-20% (w / w) buffer, 0-50% (w / w) cosolvent, and / or 0-99% (w / w) water for injection (WFI) (depending on the dose and whether it is lyophilized). Depot intramuscular formulations may also contain 0-99% (w / w) oil.

[0197] Pharmaceutical formulations are sometimes provided to patients in "patient packs", containing the entire course of treatment in one package, usually a blister pack.

[0198] The compounds of Formula I will generally be provided in unit dose form and, as such, will typically contain sufficient compound to provide a desired level of biological activity. For example, a formulation may contain 1 nanogram to 2 grams of active ingredient, e.g., 1 nanogram to 2 milligrams of active ingredient. Within these ranges, particular subranges of compound are 0.1 milligram to 2 grams of active ingredient (more usually 10 milligrams to 1 gram, e.g., 50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (e.g., 1 microgram to 10 milligrams, e.g., 0.1 milligram to 2 milligrams of active ingredient).

[0199] For oral compositions, a unit dose form may contain from 1 milligram to 2 grams, more typically from 10 milligrams to 1 gram, for example, 50 milligrams to 1 gram, for example, 100 milligrams to 1 gram, of active compound.

[0200] The active compound will be administered to a patient (e.g., a human or animal patient) in need thereof in an amount sufficient to achieve the desired therapeutic effect (an effective amount). The exact amount of the compound to be administered can be determined by a supervising physician according to standard procedures.

[0201] Process for preparing compounds of formula (I) Additional embodiments are disclosed in more detail in the following examples, which are not intended to limit the scope of the claims in any way.

[0202] General synthetic scheme

[0203] Compounds of Formula I can be prepared according to synthetic methods well known to those skilled in the art and described herein.

[0204] Scheme 1 [ka]

[0205] In Scheme 1, compounds of formula (2a) can be prepared as summarized in Scheme 1. Alkylation of amines of formula G4 can be carried out by the reaction of PG 1 represents a suitable acid protecting group such as a methyl ester, and an alkylating agent of formula G5 is used, where LG represents a suitable leaving group, typically bromide. Typically, the alkylation reaction is carried out in the presence of a base such as NaHCO3, in a solvent such as MeCN, at a temperature ranging from room temperature to 70°C, to give a tertiary amine of formula G6. The resulting ester is then converted to a protecting group PG 1 Deprotection can be achieved using conditions appropriate to the nature of the compound, typically hydrolysis of the methyl ester functionality in the presence of a nucleophilic base such as lithium hydroxide in a solvent such as MeOH, to provide acids of formula (7). An amide bond forming reaction between acids of formula G7 and amines of formula G8a in the presence of an amide coupling reagent such as HOBt or EDCI and a base such as triethylamine in a solvent such as DCM provides compounds of formula 2a.

[0206] A is a protected carboxylic acid group, and PG 2 represents a suitable acid protecting group such as a methyl ester, coupling gives an amide of formula G9b. Amide G9b can be prepared by coupling a protecting group PG 2 Further deprotection can be performed using conditions appropriate to the nature of the compound. Typically, hydrolysis of the methyl ester functionality in the presence of a nucleophilic base such as lithium hydroxide in a solvent such as MeOH provides compounds of formula 2b. See Scheme 2.

[0207] Scheme 2 [ka]

[0208] Alternatively, in Scheme 3, compounds of formula 2a can be synthesized via compounds of formula G12, as described in Scheme 3 and Scheme 4. According to Scheme 3, PG 3Compounds of formula G12 can be obtained by coupling an acid of formula G10, where represents a suitable amine protecting group such as BOC, with an amine of formula G8a in the presence of a suitable coupling agent such as HATU and a base such as N,N-diisopropylethylamine in a solvent such as MeCN or DMF to give the desired amide of formula G11. 3 Deprotection under conditions appropriate to the nature of the Boc deprotection, typically in the presence of an acid such as HCl in a suitable solvent such as 1,4-dioxane, provides the desired amine intermediate of formula G12. Alternatively, amines of formula G12 can be generated by coupling amino acid G13 with amines of formula G8a using amide bond forming conditions as described above.

[0209] Scheme 3 [ka]

[0210] Amines of formula G12 can then be benzylated by reductive amination with aldehydes of formula G14 to give the corresponding tertiary amines of formula G16. Typical reductive amination conditions use reagents such as sodium triacetoxyborohydride or sodium cyanoborohydride in the presence or absence of a cocatalyst such as glacial acetic acid or zinc chloride, in a solvent such as DCM or methanol, at temperatures ranging from room temperature to 70°C. Alternatively, tertiary amines of formula 16 can be prepared via alkylation of amines of formula G12 with alkylating agents of formula G15, where LG represents a suitable leaving group such as a halide, typically bromide. Typical alkylation conditions use a suitable base such as potassium carbonate, N,N-diisopropylethylamine, or sodium bicarbonate, in the presence or absence of sodium iodide, in a solvent such as acetonitrile or DMF, at temperatures ranging from room temperature to 80°C. The resulting compound of formula G16, where Hal is a suitable halide, typically bromide, can be converted to a compound of formula 2a by Suzuki coupling with a boronic acid or boronic ester of formula G17, where R is alkyl, such as methyl. This cross-coupling reaction is carried out in the presence of a catalyst such as tetrakistriphenylphosphinepalladium 0 or PdCl2dppf.DCM, with a base such as potassium carbonate, typically in a solvent combination such as 1,4-dioxane and water, at a reaction temperature between room temperature and 100°C, with or without microwave irradiation.

[0211] Scheme 4 [ka]

[0212] In the compounds of Scheme 3 and Scheme 4, A is a protected carboxylic acid group and PG 2 When represents a suitable acid protecting group such as a methyl ester, compounds of formula G16 and formula G17 are coupled to give compounds of formula G9b according to Scheme 5. Compounds of formula G9b can be coupled with the protecting group PG 2Deprotection can be achieved using hydrolysis of the methyl ester functionality under conditions appropriate to the nature of the compound, typically in the presence of a nucleophilic base such as lithium hydroxide in a solvent such as MeOH, to provide the carboxylic acid compound of formula 2b.

[0213] Scheme 5 [ka]

[0214] Those skilled in the art will appreciate that the reaction steps shown in Schemes 3-5 can be combined in various ways as necessary to successfully prepare the desired compound of Formula 2a. This may involve the inclusion of additional steps, such as functional group modification, protection, and / or deprotection, within the overall synthetic sequence. For example, compounds of Formula 2a can be prepared as shown in Scheme 6. Reductive amination between a secondary amine of Formula G12 and an aldehyde of Formula G19 under the conditions outlined in Scheme 4 above provides the desired tertiary amine of Formula G20. Subsequent Suzuki reaction with an appropriate aryl halide of Formula G21, where Hal is typically a bromide, using the conditions described in Scheme 4, provides compounds of Formula 2a. Subsequent deprotection of the ester of Formula 18, as described in Scheme 2, provides compounds of Formula 2a.

[0215] Scheme 6 [ka]

[0216] In the compounds of Scheme 6, A is a protected carboxylic acid group and PG 2 When represents a suitable acid protecting group such as a methyl ester, compounds of formula G20 and formula G21 are coupled to produce compounds of formula G9b according to Scheme 7. Compounds of formula G9b can be coupled with a protecting group PG 2Deprotection can be achieved using hydrolysis of the methyl ester functionality under conditions appropriate to the nature of the compound, typically in the presence of a nucleophilic base such as lithium hydroxide in a solvent such as MeOH, to provide the carboxylic acid compound of formula 2b.

[0217] Scheme 7 [ka]

[0218] In another synthetic variation, compounds of formula 2a and formula 2b can be prepared as described in Scheme 8. Morpholines of formula G12 are directly converted to compounds of formula 2a via reductive amination or alkylation reactions using the corresponding aldehydes of formula G22 or electrophiles of formula G5, respectively, under the conditions described for Scheme 4. In the compounds of Scheme 8, A is a protected carboxylic acid group and PG 2 When represents a suitable acid protecting group such as a methyl ester, subsequent deprotection of the ester of formula 2a can provide compounds of formula 2b.

[0219] Scheme 8 [ka]

[0220] The intermediate aldehyde of formula G22 and alkylating agent of formula G5 can be prepared as described in Scheme 9. Coupling of a boronic acid or boronic ester of formula G17, where R is alkyl, such as methyl, with an aldehyde of formula G14 under Suzuki conditions, as described in Scheme 2, provides an aldehyde intermediate of formula G22. Reduction of the aldehyde moiety of formula G22 with a reagent such as sodium borohydride in a solvent such as methanol provides an alcohol of formula G23. Alternatively, the alcohol of formula G23 can be prepared via reaction of a boronic acid or boronic ester of formula G17 with an aromatic halide of formula G24, where Hal is typically a bromide, in a palladium-catalyzed Suzuki reaction, as described above. The alcohol of formula G23 can be converted to a suitable alkylating agent of formula G5, where the leaving group LG is suitable for the nature of the subsequent alkylation reaction, typically a bromide. Typical bromination conditions involve treatment with 33% HBr / acetic acid at room temperature, PBr / DCM at room temperature, or SOCl / DCM at room temperature. It will be appreciated by those skilled in the art that other leaving groups can be formed, including but not limited to chloride, mesylate, and tosylate, and that other conditions will be required to synthesize such intermediates of formula G5.

[0221] Scheme 9 [ka]

[0222] In another variation of compound 2a, where U, V, W, or Z is COH, compounds of formula G27 can be synthesized as described in Scheme 10. Protected phenol compounds of formula G25, where PG is a suitable protecting group, typically methyl or benzyl, can be synthesized from morpholines of formula G12 as described in Scheme 4. Suzuki reaction between an aryl halide of formula G25, where Hal is typically a bromide, and a boronic acid or boronic ester of formula G17, under conditions described for Scheme 4, provides biaryl molecules of formula G26. A is a protected carboxylic acid group, and PG 2Deprotection of the phenol moiety and the protected acid moiety in the compound, where represents a suitable acid protecting group, provides a compound of formula G27. These protecting groups are typically methoxy and methyl ester groups, respectively, and typical methoxy deprotection conditions are boron tribromide in a solvent such as DCM at temperatures ranging from −78° C. to room temperature. Hydrolysis conditions using hydroxide base are typically used to deprotect the ester moiety.

[0223] Scheme 10 [ka]

[0224] R 5 In other variations of compound 2a, where PG is OSONH and U, V, W, or Z is H or COH, compounds of formula 29 or 32 can be prepared as described in Scheme 11. First, a phenol intermediate of formula G28 is synthesized from a morpholine intermediate of formula G12 as described herein. Sulfamoylation of the phenol group of formula G28 can be achieved under appropriate conditions, typically via treatment with excess sulfamoyl chloride in DMA, to give the final compound of formula G29. Alternatively, a phenol-protected intermediate compound of formula G30, where PG is a suitable protecting group, typically benzyl, can be synthesized from a morpholine of formula G12 as described herein. By carrying out the sulfamoylation step as described above, the compound of formula G28 is converted to a compound of formula G29. Removal of the phenol protecting group of a compound of formula G31, typically by hydrogenation of the benzyl group in the presence of a catalyst such as palladium on carbon in a solvent such as ethanol, gives the final compound of formula G32.

[0225] Scheme 11 [ka]

[0226] It should be understood that the above schemes and procedures are not intended to be limiting in any way. Indeed, the above schemes and procedures can also be used to prepare compounds of the present invention, for example, where A is a carboxylic acid isostere group. Suitable methods and protecting groups for the "A group" are well known to those skilled in the art; for example, a trityl group can be used to protect a tetrazole group. Furthermore, a compound of Formula 1 can be converted to another compound of the present invention by methods well known to those skilled in the art. Examples of synthetic procedures for converting one functional group into another can be found in standard textbooks 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 1-17, John Wiley, edited by Mary Fieser (ISBN: 0-471-58283-2).

[0227] In many of the above reactions, it may be necessary to protect one or more groups to prevent reaction from occurring at undesired locations on the molecule. Examples of protecting groups and methods for protecting and deprotecting functional groups can be found in Greene's Protective Groups in Organic Synthesis, Fifth Edition, Editor: Peter GM Wuts, John Wiley, 2014, (ISBN:9781118057483).

[0228] The compounds produced by the above methods can be isolated and purified by any of a variety of methods known to those skilled in the art, including recrystallization and chromatographic methods such as column chromatography (e.g., flash chromatography), HPLC, and SFC under normal or reverse phase conditions. [Example]

[0229] The present invention will now be described with reference to the following examples, but is not limited thereto.

[0230] Basic steps Where preparative routes are not included, relevant intermediates are commercially available. Data on intermediates are listed in Table 2. Commercially available reagents were used without further purification. Final compounds and intermediates are named using ChemDraw Professional (version 17.0.0.206(121)). Room temperature (RT) refers to approximately 20-27 °C. H NMR spectra were recorded at 400 MHz 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 = 7.26 ppm, DMSO - d = 2.50 ppm, methanol - d = 3.31 ppm. The following abbreviations are used for NMR signal multiplicity: s = singlet, br = broad, d = doublet, t = triplet, q = quartet, m = multiplet. Coupling constants are listed as J values and are measured in Hz. NMR and mass spectrometry results were corrected for background peaks. Chromatography refers to column chromatography performed using 60-120 mesh or 40-633 μm, 60 Å silica gel under nitrogen pressure (flash chromatography). Microwave-mediated reactions were performed in a Biotage Initiator or CEM Discover microwave reactor.

[0231] LC / MS analysis LC / MS analysis of compounds was carried out under electrospray conditions using the following equipment and method:

[0232] LC / MS Method A

[0233] Equipment: ACQUITY UPLC equipped with a photodiode array detector and a QDa mass detector; Column: ACQUITY C-18, 1.6 micron, 50x1.6 mm; Gradient [time (min) / % B in solvent 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.

[0234] LC / MS Methods B and C

[0235] Apparatus: HP 1100 with G1315A DAD, Waters Micromass ZQ; Column: Phenomenex Gemini-NX C-18, 3 micron, 2.0 x 30 mm; Gradient for Method B [time (min) / % B in solvent A]: 0.00 / 2, 0.10 / 2, 2.50 / 95, 3.50 / 95; Gradient for Method C [time (min) / % B in solvent A]: 0.00 / 2, 0.01 / 2, 8.40 / 95, 10.00 / 95; Solvents: Solvent A = 2.5 L HO + 2.5 mL 28% ammonia in HO; Solvent B = 2.5 L MeCN + 135 mL HO + 2.5 mL 28% ammonia in HO. Injection volume 1 μL; UV detection 230–400 nm; mass detection 130–800 amu; column temperature 45°C; flow rate 1.5 mL / min.

[0236] LC / MS Method D

[0237] Instrument: HP1100 with G1315A DAD and Waters Micromass ZQ column; Column: Phenomenex Kinetex C-18, 5 micron, 2.1 x 50 mm; Gradient [Time (min) / % B in Solvent A]: 0.00 / 2, 0.01 / 2, 8.40 / 95, 10.00 / 95; Solvents: Solvent A = 0.1% formic acid in 10 mM aqueous ammonium formate; Solvent B = acetonitrile / water / formic acid (95:4.9:0.1). Injection volume: 1 μL; UV detection: 230-400 nm; Mass detection: 130-800 amu; Column temperature: 45 °C; Flow rate: 1.5 mL / min.

[0238] LC / MS Method E

[0239] Equipment: ACQUITY UPLC equipped with a photodiode array detector and a QDa mass detector; Column: ACQUITY C-18, 1.6 micron, 50x1.6 mm; Gradient [time (min) / % B in solvent 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.

[0240] LC / MS Methods F and G

[0241] Instrument: ACQUITY H-Class equipped with a photodiode array detector and a QDa mass detector; Column: ACQUITY C-18, 1.6 micron, 50x1.6 mm; Method F gradient [time (min) / % B in solvent 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) / % B in solvent 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.

[0242] LC / MS method H

[0243] Instrumentation: Agilent 1260 Infinity LC with diode array detector, Agilent 6120B Single Quadrupole MS with API-ES source; Column: Restek Penta Fluoro Phenyl Propyl, 3 micron, 2.1 x 30 mm. Gradient [time (min) / % B in Solvent A]: 0.00 / 2, 0.1 / 2, 8.4 / 95, 10 / 95, 10.1 / 2, 12 / 2; Solvents: Solvent A = water (2.5 L) + 2.5 mL formic acid; Solvent B = MeCN (2.5 L) + 125 mL water and 2.5 mL formic acid. Injection volume: 0.5 μL; UV detection: 190-400 nm; Mass detection: 130-800 amu; Column temperature: 40 °C; Flow rate: 1.5 mL / min.

[0244] LC / MS Method I

[0245] Apparatus: Shimadzu 2020 series with diode array detector; Column: Agilent Poroshell 120 EC C-18, 2.7 micron, 4.6 x 50 mm; Gradient [time (min) / % B in solvent A]: 0.00 / 15, 1.00 / 15, 4.00 / 100, 4.50 / 100, 4.51 / 15, 5.00 / 15; Solvents: 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.

[0246] HPLC purification Where stated, intermediates or final compounds were purified by reverse phase preparative HPLC conditions using the equipment and methods shown below:

[0247] HPLC purification method A

[0248] Gilson semi-preparative HPLC system equipped with a 321 pump, a 171 diode array detector, and a GX-271 liquid handler, with Gilson Trilution software.

[0249] Preparative HPLC: [reverse phase (Phenomenex Kinetix C18, 100 x 30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: 5% → 35% solvent B over 10 min, 100% solvent B over 2 min; solvent A: water containing 0.1% TFA. Solvent B: MeCN].

[0250] HPLC purification method B

[0251] Gilson semi-preparative HPLC system equipped with a 321 pump, a 171 diode array detector, and a GX-271 liquid handler, with Gilson Trilution software.

[0252] Preparative HPLC: [reverse phase (Gemini-NX C18, 100 × 30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: 5% → 35% solvent B over 10 min, 100% solvent B over 2 min; solvent A: 28% ammonia in water with 0.2%. Solvent B: MeCN].

[0253] HPLC purification method C

[0254] Gilson semi-preparative HPLC system equipped with dual piston pumps 331 and 332, a 171 diode array detector, and a GX-271 liquid handler, and Gilson Trilution software.

[0255] Preparative HPLC: [reverse phase (Gemini-NX C18, 100 × 30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: 5% → 35% solvent B over 10 min, 100% solvent B over 2 min; solvent A: 28% ammonia in water with 0.2%. Solvent B: MeCN].

[0256] HPLC purification method D

[0257] Gilson semi-preparative HPLC system equipped with dual piston pumps 331 and 332, a 171 diode array detector, and a GX-271 liquid handler, and Gilson Trilution software.

[0258] Preparative HPLC: [reverse phase (Phenomenex Kinetix C18, 100 x 30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: 5% → 35% solvent B over 10 min, 100% solvent B over 2 min; solvent A: water containing 0.1% TFA. Solvent B: MeCN].

[0259] HPLC Purification Method E

[0260] Shimadzu preparative HPLC equipped with an LC-20Ap pump, an SPD-20A diode array detector, and a CBM-20A controller.

[0261] Preparative HPLC: [reverse phase (Agilent Zorbax, 10 mL / min, gradient of solvent B in solvent A: 0% → 23% solvent B over 14 min, 98% solvent B over 2 min; solvent A: water containing 0.1% TFA. Solvent B: MeCN].

[0262] HPLC purification method F

[0263] Shimadzu preparative HPLC equipped with an LC-20Ap pump, SPD-20A diode array detector and Labsolutions software.

[0264] Preparative HPLC: [reverse phase (Agilent 10C18, 250 x 21 mm, 20 mL / min, gradient of solvent B in solvent A: 5% → 80% solvent B; solvent A: water containing 0.1% TFA. Solvent B: MeCN].

[0265] HPLC purification method G

[0266] Shimadzu preparative HPLC equipped with an LC-20Ap pump, an SPD-20A diode array detector, and a CBM-20A controller.

[0267] Preparative HPLC: [reverse phase (Agilent, 250x9.4 mm, 55 μm, 9 mL / min, gradient of solvent B in solvent A: 28% → 45% solvent B (over 18 min), 45% → 98% solvent B (2 min), 28% solvent B (2 min); solvent A: 5 mM aqueous ammonium bicarbonate. Solvent B: MeCN].

[0268] Synthetic procedure for intermediates Representative Route 1 for the Preparation of Intermediate 1: Methyl 4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate Hydrochloride (Intermediate 1) [ka]

[0269] Step (i): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (5.10 g, 22.1 mmol) and (S)-methyl 4-(1-aminoethyl)benzoate 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 time, the mixture was cooled to 0 °C, and N,N-diisopropylethylamine (11.9 mL, 66.2 mmol) was added. The mixture was allowed to warm 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 (twice) with EtOAc. The combined organic layers were dried over Na2SO4 and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of EtOAc in hexanes (0% to 72%) to give tert-butyl (R)-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 + ), 2.17 min, UV detectable.

[0270] Step (ii): To a solution of tert-butyl (R)-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 4 N HCl / 1,4-dioxane (40 mL) at room temperature under a nitrogen atmosphere. The mixture was stirred for 3 h, and then the solvent was removed under reduced pressure. The crude material was triturated with EtO to give intermediate 1, methyl 4-((S)-1-((R)-morpholine-3-carboxamido)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 + ), 2.17 min, UV detectable.

[0271] Representative Route 2 for the Preparation of Intermediate 2: 3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (Intermediate 2) [ka]

[0272] Step (i): A solution of 4-bromo-3-methylbenzoic acid (30 g, 0.14 mol) in SOCl2 (250 mL) was stirred at 70 °C for 15 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in DCM (50 mL), cooled to 0 °C, and aqueous ammonium hydroxide (40 mL) was added dropwise. After stirring the mixture at room temperature for 30 min, the solid was filtered off and recrystallized from EtOH (250 mL) to give 4-bromo-3-methylbenzamide (21.0 g, 0.10 mol, 70% yield) as a yellow solid. (LC / MS Method I): m / z 214 [M+H] + (ES + ), 2.76 min, UV detectable.

[0273] Step (ii): A mixture of 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 (700 mg, 0.96 mmol) in 1,4-dioxane (150 mL) was heated to 105 °C under a nitrogen atmosphere for 15 h. The mixture was cooled to room temperature, and the solvent was removed in vacuo. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 10% to 30% EtOAc in petroleum ether to afford intermediate 2, 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8.40 g, 32.2 mmol, 96% yield), as a yellow solid. (LC / MS Method I): m / z 262 [M+H] + (ES + ), 4.84 min, UV detectable.

[0274] Representative Route 3 for the Preparation of Intermediate 3: Methyl 4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (Intermediate 3) [ka]

[0275] 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 and then partitioned between EtoAc and water. The organic layer was separated, and the aqueous layer was further extracted (1×) with EtOAc. The combined organic layers were dried over Na2SO4 and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 0% to 52% EtOAc in hexanes to give intermediate 3, methyl 4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (0.80 g, 2.74 mmol, 49% yield) as a viscous liquid. (LC / MS Method A): m / z 293 [M+H] + (ES + ), 1.33 min, UV detectable.

[0276] Representative Route 4 for the Preparation of Intermediate 4: 3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (Intermediate 4) [ka]

[0277] Step (i): To a solution of 4-bromo-3-methylbenzenesulfonyl chloride (10.0 g, 37.1 mmol) in THF (75 mL) at 0 °C, aqueous ammonia (75 mL) was added. The mixture was stirred at the same temperature for 5 h and then quenched by adding 1 N HCl. The mixture was extracted with DCM (twice), and the combined organic layers 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 + ), 3.27 min, UV detectable.

[0278] Step (ii): A mixture of 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 (900 mg, 1.23 mmol) in 1,4-dioxane (100 mL) was heated to 105 °C under a nitrogen atmosphere for 15 h. The mixture was cooled to room temperature, and the solvent was removed in vacuo. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 20% to 25% EtOAc in petroleum ether to give intermediate 4, 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (6.32 g, 21.3 mmol, 57% yield) as a white solid. (LC / MS Method I): m / z 298 [M+H] + (ES + ), 3.64 min, UV detectable.

[0279] Representative Route 5 for the Preparation of Intermediate 5: Methyl 5-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylate Hydrochloride (Intermediate 5) [ka]

[0280] Step (i): To a mixture of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (310 mg, 1.34 mmol) in DMF (5.5 mL) was added N,N-diisopropylethylamine (1.16 mL, 6.7 mmol), HATU (613 mg, 1.61 mmol), and methyl 5-(aminomethyl)-2-pyridinecarboxylate dihydrochloride (405 mg, 1.61 mmol). The mixture was stirred at room temperature for 18 h and then diluted with EtOAc and water. The organic layer was separated, washed with brine (twice), dried by passing through a hydrophobic frit, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of MeOH (0% to 5%) in DCM to give (R)-tert-butyl 3-(((6-(methoxycarbonyl)pyridin-3-yl)methyl)carbamoyl)morpholine-4-carboxylate (537 mg, 1.42 mmol, quantitative yield) as a pale orange solid. (LC / MS Method B): m / z 380 [M+H] + (ES + ), 1.67 min, UV detectable.

[0281] Step (ii): To a solution of tert-butyl (R)-3-(((6-(methoxycarbonyl)pyridin-3-yl)methyl)carbamoyl)morpholine-4-carboxylate (510 mg, 1.34 mmol) in 1,4-dioxane (5 mL) was added 4 M HCl / dioxane (5 mL, 1.34 mmol). The mixture was stirred at room temperature for 2 hours and then concentrated to give intermediate 5, methyl 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 + ), 0.37 min, UV detectable.

[0282] Representative Route 6 for 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) [ka]

[0283] 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 gas at room temperature for 30 minutes. PdCl(dppf).DCM (1.63 g, 2.00 mmol) was then added, and the reaction mixture was heated to 80 °C and maintained 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. The combined organic layers were dried over NaSO and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) using 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 + ), 1.09 min, UV detectable.

[0284] 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) at 0 °C was added sodium borohydride (440 mg, 11.8 mmol). The reaction mixture was stirred at room temperature for 2 h and then partitioned between EtOAc and saturated aqueous NaHCO3. The organic layer was separated and the aqueous layer was further extracted (twice) with EtOAc. The combined organic layers were dried over Na2SO4 and concentrated to give 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 + ), 0.94 min, UV detectable.

[0285] Step (iii): 3'-Hydroxy-5'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (1.36 g, 5.29 mmol) was dissolved in 33% HBr / acetic acid (12 mL) at room temperature. The mixture was stirred at room temperature for 16 hours and then partitioned between saturated aqueous NaHCO3 and EtOAc. The organic layer was separated, and the aqueous layer was further extracted (twice) with EtOAc. The combined organic layers were dried over Na2SO4 and concentrated to give 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 + ), 1.23 min, UV detectable.

[0286] Representative Route 7 for the Preparation of Intermediate 7: 3-Benzyloxy-5-bromo-benzaldehyde (Intermediate 7) [ka]

[0287] 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 h and then partitioned between EtOH and water. The organic layer was separated, washed successively with water and brine, dried by passing through a hydrophobic frit, and concentrated. The residue was purified by flash column chromatography (normal phase, silica) using a gradient of EtO in isohexane (0% → 10%) to give intermediate 7, 3-benzyloxy-5-bromo-benzaldehyde (736 mg, 2.53 mmol, quantitative yield) as a dark red oil. (LC / MS Method B): m / z 292 [M+H] + (ES + ), 2.43 min, UV detectable.

[0288] Representative Route 8 for the Preparation of Intermediate 8: Methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (Intermediate 8) [ka]

[0289] 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 / 1,4-dioxane (10 mL) was added, and the reaction mixture was heated to reflux for 8 hours. The mixture was cooled and concentrated, and the resulting residue was triturated with diethyl ether to give (S)-methyl 4-(1-aminoethyl)-2-methoxybenzoate hydrochloride (2.10 g, 8.6 mmol, 83% yield) as a white solid. 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).

[0290] Step (ii): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (3.60 g, 15.5 mmol) and (S)-methyl 4-(1-aminoethyl)-2-methoxybenzoate 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 and then partitioned between EtOAc and water. The organic layer was separated, and the aqueous layer was further extracted (twice) with EtOAc. The combined organic layers were dried over Na2SO4 and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 0% to 48% EtOAc in hexanes to give tert-butyl (R)-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 + ), 1.78 min, UV detectable.

[0291] Step (iii): To a solution of tert-butyl (R)-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) at room temperature, 4 M HCl / 1,4-dioxane (60 mL) was added, and the mixture was stirred at room temperature for 2 h. After this time, the mixture was partitioned between EtOAc and saturated aqueous NaHCO. The organic layer was separated, and the aqueous layer was further extracted (twice) with EtOAc. The combined organic layers were dried over NaSO and concentrated to give methyl 2-methoxy-4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (3.20 g, 9.93 mmol, 68% yield) as a white solid. (LC / MS Method E): m / z 323 [M+H] + (ES +), 0.99 min, UV detectable.

[0292] Step (iv): To a solution of methyl 2-methoxy-4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (3.20 g, 9.93 mmol) in DCM at −78 °C, BBr (1 M in DCM, 32 mL) was added dropwise, and the mixture was stirred at the same temperature for 1 h. After this time, the mixture was partitioned between EtOAc and saturated aqueous NaHCO. The organic layer was separated, and the aqueous layer was further extracted (twice) with EtOAc. The combined organic layers were dried over NaSO and concentrated. The residue was purified by flash column chromatography (reverse-phase, C18) using a gradient of MeCN in water (0 to 33%) to give intermediate 8, methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (2.30 g, 7.46 mmol, 75% yield) as a white solid. (LC / MS Method E): m / z 309 [M+H] + (ES + ), 1.34 min, UV detectable.

[0293] Representative Route 9 for the Preparation of Intermediate 11: 3'-(Bromomethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 11) [ka]

[0294] 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 (224 mg, 0.306 mmol) under a nitrogen atmosphere. The mixture was heated to 85 °C for 2.5 h and then concentrated under reduced pressure. The residue was partitioned between EtOAc and water, and the organic layer was separated, dried over NaSO, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 5% to 23% EtOAc 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 + ), 2.80 min, UV detectable.

[0295] Step (ii): To a solution of 3'-(hydroxymethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (300 mg, 1.24 mmol) in DCM (30 mL) at 0 °C, PBr (503 mg, 1.86 mmol) was added, and the mixture was stirred at room temperature for 2 h. After this time, the mixture was poured into ice-cold water and extracted (1×) with DCM. The organic layer was dried over NaSO and concentrated to give 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 + ), 3.60 min, UV detectable.

[0296] Representative Route 10 for the Preparation of Intermediate 12 and Intermediate 30: Methyl 4-(1-aminoethyl)-2,6-difluorobenzoate Hydrochloride (Intermediate 12 and Intermediate 30 are unidentified separated enantiomers) [ka]

[0297] Step (i): To a solution of 2,6-difluoro-4-formylbenzonitrile (5.00 g, 29.9 mmol) and (R)-tert-butanesulfinamide (3.63 g, 29.9 mmol) in anhydrous THF (60 mL) was added Ti(OEt) (12.5 mL, 59.8 mmol) dropwise. The mixture was stirred at room temperature for 16 h and then poured into brine. The resulting suspension was filtered through Celite and washed with EtOAc. The filtrate was washed with brine, and the organic layer was separated. The aqueous layer was further extracted with EtOAc (twice), and the combined organic layers were dried over NaSO and concentrated. The residue was recrystallized from EtOAc / hexane to give (R)-N-(4-cyano-3,5-difluorobenzylidene)-2-methylpropane-2-sulfinamide (5.00 g, 18.5 mmol, 62% yield) as a white solid. (LC / MS Method I): m / z 271 [M+H] + (ES + ), 3.93 min, UV detectable.

[0298] Step (ii): To a solution of (R)-N-(4-cyano-3,5-difluorobenzylidene)-2-methylpropane-2-sulfinamide (5.50 g, 20.4 mmol) in anhydrous DCM (100 mL) at −78° C. under a nitrogen atmosphere, methylmagnesium bromide (3 M in EtO, 13.6 mL) was added dropwise. The mixture was stirred at the same temperature for 1 h, then allowed to warm to room temperature and stirred at room temperature for 1 h. The mixture was cooled to 0° C. and quenched by the slow addition of saturated aqueous NH4Cl (20 mL). The organic layer was separated, and the aqueous layer was extracted with EtOAc (3×). The combined organic layers were dried over NaSO and concentrated, and the residue was recrystallized from EtOAc / hexane to give a diastereomeric mixture of (R)-N-(1-(4-cyano-3,5-difluorophenyl)ethyl)-2-methylpropane-2-sulfinamide (5.00 g, 17.5 mmol, 86% yield) as a yellow solid. (LC / MS Method I): m / z 287 [M+H] + (ES +), 3.48 min, UV detectable.

[0299] Step (iii): A suspension of the diastereomeric mixture of (R)—N-(1-(4-cyano-3,5-difluorophenyl)ethyl)-2-methylpropane-2-sulfinamide (350 mg, 1.22 mmol) and NaOH (147 mg, 3.67 mmol) was heated to 80° C. for 5 h, then cooled to room temperature, diluted with saturated aqueous citric acid, and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over NaSO, and concentrated to give the diastereomeric mixture of 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoic acid (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 + ), 2.60 min, UV detectable.

[0300] Step (iv): To a solution of the diastereomeric mixture of 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoic acid (2.66 g, 8.71 mmol) in MeOH (50 mL) 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) at 0° C. The mixture was stirred at room temperature for 15 h, then diluted with water and extracted with DCM (×3). The combined organic layers were washed with brine, dried over NaSO, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 10% to 30% EtOAc in petroleum ether to give diastereomer 1 (1.00 g, 3.13 mmol, 36% yield), methyl 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoate, as a yellow oil in the first eluting spot, and diastereomer 2 (665 mg, 2.08 mmol, 24% yield), methyl 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoate, as a yellow oil in the later eluting spot. Diastereomer 1: (LC / MS Method I): m / z 320 [M+H] + (ES + ), 3.33 min, UV detectable. Diastereomer 2: (LC / MS Method I): m / z 320 [M+H] + (ES + ), 3.24 min, UV detectable.

[0301] Step (v): To two separate solutions of each purified diastereomer, methyl 4-(1-(((R)-tert-butylsulfinyl)amino)ethyl)-2,6-difluorobenzoate (60 mg, 0.19 mmol) in MeOH (3 mL), 4 M HCl / 1,4-dioxane was added, and the mixture was stirred at room temperature for 1 h. The solution was concentrated to give intermediate 12 (47 mg, 0.19 mmol, 98% yield; (LC / MS Method I): m / z 216 [M+H-HCl]). + (ES +), 1.66 min, UV detectable. ) as a yellow solid or intermediate 30 (45 mg, 0.18 mmol, 94% yield; (LC / MS Method I): m / z 315 [M+H-HCl] + (ES + ), 1.49 min, UV detectable. ) was obtained as a yellow solid.

[0302] Representative Route 11 for the Preparation of Intermediate 13: 4-Bromo-3-((tert-butyldimethylsilyl)oxy)benzenesulfonamide (Intermediate 13) [ka]

[0303] Step (i): To a solution of 4-bromo-3-methoxybenzenesulfonamide (1.00 g, 3.75 mmol) in DCM (10 mL) at 0 °C was added BBr (1 M in DCM, 7.10 mL, 7.10 mmol). The mixture was stirred at room temperature for 16 h and then partitioned between EtOAc and saturated aqueous NaHCO. The organic layer was separated, and the aqueous layer was further extracted (twice) with EtOAc. The combined organic layers were dried over NaSO and concentrated under reduced pressure, and the residue was purified by flash column chromatography (reverse-phase, C18) using a gradient of MeCN in water (0 to 26%) to give 4-bromo-3-hydroxybenzenesulfonamide (0.70 g, 2.94 mmol, 74% yield) as an off-white solid. (LC / MS Method E): m / z 252 [M+H] + (ES + ), 1.43 min, UV detectable.

[0304] 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 min, after which tert-butyldimethylsilyl chloride (180 mg, 1.19 mmol) was added at the same temperature. The mixture was allowed to warm to room temperature and stirred for 1 h before being partitioned between EtOAc and water. The organic layer was separated, and the aqueous layer was further extracted (twice) with EtOAc. The combined organic layers were dried over NaSO and concentrated to give 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. 1 H 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).

[0305] Representative Route 12 for the Preparation of Intermediate 14: 3-(4-Carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (Intermediate 14) [ka]

[0306] 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 (1.12 g, 1.53 mmol) under a nitrogen atmosphere, 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) using a gradient of MeOH (5% to 10%) 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 + ), 0.80 min, UV detectable.

[0307] 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 (5.83 g, 49.0 mmol), and the mixture was stirred at room temperature for 1 h, then filtered and concentrated. The crude residue was purified by flash column chromatography (normal phase, silica) using a gradient of MeOH (5% to 10%) in DCM to give 4-(5-(chloromethyl)pyridin-3-yl)-3-methylbenzamide (1.0 g, 3.85 mmol, 78% yield) as a yellow solid. (LC / MS Method I): m / z 261 [M+H] + (ES + ), 2.73 min, UV detectable.

[0308] 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 metachloroperbenzoic acid (664 mg, 3.84 mmol). The mixture was stirred at room temperature for 1 h and then concentrated under reduced pressure. The crude residue was purified by flash column chromatography (normal phase, silica) using a gradient of MeOH (5% to 7%) in DCM to give intermediate 14, 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (500 mg, 1.81 mmol, 94% yield) as a pale yellow solid. (LC / MS Method I): m / z 277 [M+H] + (ES + ), 2.30 min, UV detectable.

[0309] Representative Route 13 for the Preparation of Intermediate 18: Ethyl 6-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-3-carboxylate Hydrochloride (Intermediate 18) [ka]

[0310] 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 ethyl 6-(aminomethyl)nicotinate hydrochloride (247 mg, 1.14 mmol). The mixture was stirred at room temperature for 18 h and then partitioned between EtoAc and water. The organic layer was separated, washed with brine, dried by passing through a hydrophobic frit, and concentrated. The crude residue was purified by flash column chromatography (normal phase, silica) using 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 pale orange gum. (LC / MS Method B): m / z 394 [M+H] + (ES + ), 1.90 min, UV detectable.

[0311] Step (ii): To a solution of tert-butyl (3R)-3-[(5-ethoxycarbonyl-2-pyridyl)methylcarbamoyl]morpholine-4-carboxylate (379 mg, 0.96 mmol) in 1,4-dioxane (4 mL), 4 M HCl / 1,4-dioxane (4 mL, 1.34 mmol) was added. The mixture was stirred at room temperature for 2.5 hours and then 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 + ), 1.38 min, UV detectable.

[0312] Representative Route 14 for the Preparation of Intermediate 20: Methyl 2,6-difluoro-4-[[[(3R)-morpholine-3-carbonyl]amino]methyl]benzoate Hydrochloride (Intermediate 20) [ka]

[0313] 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)methanamine hydrochloride (890 mg, 3.44 mmol). The mixture was stirred at room temperature for 18 h and then partitioned between EtoAc and water. The organic layer was separated, washed with brine, dried by passing through a hydrophobic frit, and concentrated. The crude residue was purified by flash column chromatography (normal phase, silica) using a gradient of 20% to 80% EtOAc 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 + ), 2.18 min, UV detectable.

[0314] 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 a CO atmosphere for 20 h. The mixture was filtered through Celite and concentrated. The residue was partitioned between EtoAc and water, and the organic layer was separated, washed with brine, dried over MgSO, and concentrated. The crude residue was purified by flash column chromatography (normal phase, silica) using a gradient of MeOH (0% → 10%) in DCM, followed by further purification by preparative HPLC [Purification Method B; 40 → 70% Solvent B / 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 + ), 2.02 min, UV detectable.

[0315] Step (iii): To a stirred solution of tert-butyl (R)-3-((3,5-difluoro-4-(methoxycarbonyl)benzyl)carbamoyl)morpholine-4-carboxylate (302 mg, 0.73 mmol) in 1,4-dioxane (3 mL) was added 4 M HCl / 1,4-dioxane (3 mL). The mixture was stirred at room temperature for 2.5 hours and then 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 + ), 1.38 min, UV detectable.

[0316] Route 15 representative of the preparation of intermediate 24: (4-carbamoyl-2-hydroxyphenyl)boronic acid (intermediate 24) [ka]

[0317] 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 gas for 30 minutes. Pd(dppf)Cl2 (43 mg, 0.05 mmol) was added, 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) using a gradient of MeCN (0 to 2%) in water 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 + ), 0.75 min, UV detectable.

[0318] Route 16 representative of the preparation of Intermediate 32 and Intermediate 33: (R)-4-(1-(morpholine-3-carboxamido)cyclopropyl)benzoate methyl hydrochloride (Intermediate 32) and (R)-4-(1-(4-(3-bromo-4-hydroxybenzyl)morpholine-3-carboxamido)cyclopropyl)benzoate methyl (Intermediate 33) [ka]

[0319] 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, followed by stirring at room temperature for 6 hours. The mixture was partitioned between EtOAc and water, and the organic layer was separated. The aqueous layer was further extracted with EtOAc (twice), and the combined organic layers were dried over Na2SO4 and concentrated. The crude residue was purified by flash column chromatography (normal phase, silica) using a gradient of 0% to 40% EtOAc in hexanes to give (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 + ), 2.22 min, UV detectable.

[0320] Step (ii): (R)-tert-Butyl 3-((1-(4-(methoxycarbonyl)phenyl)cyclopropyl)carbamoyl)morpholine-4-carboxylate (5.50 g, 13.6 mmol) was dissolved in 1,4-dioxane (55 mL) under a nitrogen atmosphere, and 4 N HCl / 1,4-dioxane (55 mL) was added at room temperature. The mixture was stirred at room temperature for 8 hours and then concentrated under reduced pressure. The crude material was triturated with EtO to give intermediate 32, methyl (R)-4-(1-(morpholine-3-carboxamido)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 + ), 1.07 min, UV detectable.

[0321] Step (iii): To a solution of intermediate 32, methyl (R)-4-(1-(morpholine-3-carboxamido)cyclopropyl)benzoate hydrochloride (1.36 g, 4.00 mmol) and 3-bromo-4-hydroxybenzaldehyde (0.80 g, 4.00 mmol) in MeOH (8 mL) at room temperature, ZnCl (0.27 g, 2.00 mmol) was added, and the mixture was stirred at 70 °C for 16 h. The mixture was cooled to 0 °C, and sodium cyanoborohydride (0.75 g, 12.0 mmol) was added in portions, and the mixture was stirred at room temperature for 12 h. After this time, the reaction was partitioned between EtOAc and saturated aqueous NaHCO, and the organic layer was separated. The aqueous layer was further extracted with EtOAc (twice), and the combined organic layers were dried over NaSO and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) using a gradient of MeCN (0 to 65%) in water to give intermediate 33, methyl (R)-4-(1-(4-(3-bromo-4-hydroxybenzyl)morpholine-3-carboxamido)cyclopropyl)benzoate (0.35 g, 0.72 mmol, 18% yield) as a brown solid. (LC / MS Method E): m / z 490 [M+H] + (ES + ), 1.46 min, UV detectable.

[0322] Route 17 representative of the preparation of intermediate 35: (R)-N-((S)-1-(4-cyanophenyl)ethyl)morpholine-3-carboxamide hydrochloride (intermediate 35) [ka]

[0323] Step (i): To a solution of (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (0.50 g, 2.16 mmol) in acetonitrile (7 mL), (S)-4-(1-aminoethyl)benzonitrile hydrochloride (0.39 g, 2.16 mmol) was added, followed by HATU (1.23 g, 3.24 mmol), and the reaction mixture was stirred at room temperature for 30 min. The mixture was then cooled to 0 °C, and N,N-diisopropylethylamine (1.13 mL, 6.49 mmol) was added. The mixture was allowed to warm to room temperature with stirring for 4 h. 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 dried over Na SO and concentrated under reduced pressure. The crude product was purified by gradient flash column chromatography (reverse phase, C18 silica) in which the product was eluted with 0% to 70% acetonitrile / 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 + ), 2.29 min, UV detectable.

[0324] Step (ii): To a solution of tert-butyl (R)-3-(((S)-1-(4-cyanophenyl)ethyl)carbamoyl)morpholine-4-carboxylate (0.14 g, 0.40 mmol) in DCM (3 mL) at 0 °C, 4 N HCl / dioxane (2 mL) was added, and the reaction mixture was warmed and stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure to give the crude product, which was triturated with diethyl ether (3 × 5 mL) to give intermediate 35, (R)-N-((S)-1-(4-cyanophenyl)ethyl)morpholine-3-carboxamide hydrochloride (0.113 g, 98%) as an off-brown solid. (LC / MS Method A): m / z 260 [M+H-HCl] + (ES + ), 0.81 min, UV detectable.

[0325] Intermediate 10: 4-((S)-1-((R)-4-((4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Intermediate 10) 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 is obtained. (LC / MS Method E): m / z 475 [M+H] + (ES + ), 1.45 min, UV detectable.

[0326] Synthetic Procedures for Compounds of Formula I

[0327] Example 1: Synthesis of 4-((S)-1-((R)-4-((5-(4-carbamoyl-2-methylphenyl)pyridin-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 1) [ka]

[0328] Step (i): To a solution of intermediate 1, methyl 4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoate 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 h, and then sodium triacetoxyborohydride (338 mg, 1.6 mmol) was added. The mixture was stirred at room temperature for 17 h, then partitioned between DCM and 1 M aqueous NaOH. The organic layer was separated, and the aqueous layer was extracted with DCM (3×). The combined organic layers were washed with water, then brine, dried by passing through a hydrophobic frit, and concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 100% EtOAc in isohexane to give methyl 4-[(1S)-1-[[(3R)-4-[(5-bromo-3-pyridyl)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 + ), 1.97 min, UV detectable.

[0329] 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 organic layer was separated, washed with water and then brine, dried by passing through a hydrophobic frit, and concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of MeOH (0% to 10%) in DCM to give methyl 4-[(1S)-1-[[(3R)-4-[[5-(4-carbamoyl-2-methyl-phenyl)-3-pyridyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (143.2 mg, 0.27 mmol, 85% yield) as a pale yellow foam. (LC / MS Method B): m / z 517 [M+H] + (ES + ), 1.81 min, UV detectable.

[0330] Step (iii): A mixture of methyl 4-[(1S)-1-[[(3R)-4-[[5-(4-carbamoyl-2-methyl-phenyl)-3-pyridyl]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 h. 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-pyridyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (28.5 mg, 0.06 mmol, 20% yield) as a white solid. 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). COO H No proton was observed. (LC / MS Method D): m / z 503 [M+H] + (ES + ), 1.67 min, UV detectable.

[0331] Example 5: Synthesis of 4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 5) [ka]

[0332] Step (i): Intermediate 3, methyl 4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (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) at room temperature. 1-Bromo-3-(bromomethyl)benzene (280 mg, 1.12 mmol) was added, and the reaction mixture was stirred at 80 °C for 1 h. The mixture was cooled and partitioned between EtoAc and water. The organic layer was separated, and the aqueous layer was further extracted with EtOAc. The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by flash column chromatography (normal phase, silica) using a gradient of 0 to 68% EtOAc in hexanes to give methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-carboxamido)ethyl)benzoate (400 mg, 0.87 mmol, 78% yield) as an off-white solid. (LC / MS Method A): m / z 461 [M+H] + (ES + ), 2.16 min, UV detectable.

[0333] Step (ii): Methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-carboxamido)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 gas at room temperature for 30 minutes. PdCl(dppf)DCM (0.07 g, 0.09 mmol) was added, and the reaction mixture was heated to 80 °C and maintained for 3 hours. After this time, the mixture was cooled and partitioned between EtOAc and water. The organic layer was separated, and the aqueous layer was further extracted with EtOAc. The combined organic layers were dried over NaSO and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) using 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-carboxamido)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 + ), 1.90 min, UV detectable.

[0334] Step (iii): Methyl 4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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 h and then partitioned between EtOAc and water. The organic layer was separated, and the aqueous layer was acidified to pH 1 with 4 N aqueous HCl. The acidic aqueous layer was extracted with EtOAc (twice), and the combined organic layers were dried over Na2SO4 and concentrated. 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-carboxamido)ethyl)benzoic acid (68 mg, 0.13 mmol, 44% yield) as a white solid. 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). (LC / MS method A): m / z538[M+H] + (ES + ), 1.65 min, UV detectable.

[0335] 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) [ka]

[0336] Step (i): To a solution of intermediate 5, methyl 5-[[[(3R)-morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylate hydrochloride (218 mg, 0.69 mmol) in DCM (3.2 mL), 3-bromobenzaldehyde (0.16 mL, 1.38 mmol) was added. The mixture was stirred at room temperature for 1 h, and then sodium triacetoxyborohydride (307 mg, 1.45 mmol) was added. The mixture was stirred at room temperature for an additional 17 h and then concentrated under reduced pressure. The residue was suspended in saturated aqueous NaHCO3 and extracted with EtOAc (3 times). The combined organic phases were washed with water and brine, dried by passing through a hydrophobic frit, and concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) using 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 + ), 1.92 min, UV detectable.

[0337] Step (ii): A mixture of methyl 5-[[[(3R)-4-[(3-bromophenyl)methyl]morpholine-3-carbonyl]amino]methyl]pyridine-2-carboxylate (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 h. The mixture was diluted with water and EtOAc, and the organic layer was separated. The aqueous layer was adjusted to pH 5 using concentrated HCl and then extracted with EtOAc. Both the organic and pH 5 aqueous phases were concentrated under reduced pressure 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. 1 H NMR (CD3OD, 400 MHz): 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, 2H), 4.53-4.61 (m, 3H), 7.30 (d, J = 7.9 Hz, 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). No exchangeable protons were observed. (LC / MS method C): m / z489[M+H] + (ES + ), 1.88 min, UV detectable.

[0338] Example 10: 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 10) [ka]

[0339] Step (i): To a solution of intermediate 1, methyl 4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate hydrochloride (1.14 g, 3.47 mmol) and N,N-diisopropylethylamine (1.81 mL, 10.4 mmol) in MeCN (11 mL) at room temperature, intermediate 6, 3′-(bromomethyl)-5′-hydroxy-2-methyl-[1,1′-biphenyl]-4-carboxamide (1.11 g, 3.47 mmol) was added, and the mixture was heated to 70° C. for 5 h. 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 dried over NaSO and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) using 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-carboxamido)ethyl)benzoate (1.62 g, 3.05 mmol, 88% yield) as a brown solid. (LC / MS Method F): m / z 532 [M+H] + (ES + ), 1.11 min, UV detectable.

[0340] Step (ii): Methyl 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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, acidified to pH 2 with 1N aqueous HCl, and then extracted with EtOAc (3 times). The combined organic layers were dried over Na2SO4 and concentrated. 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-carboxamido)ethyl)benzoic acid (1.06 g, 2.05 mmol, 68% yield) as an off-white solid. 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). (LC / MS method G): m / z518[M+H] + (ES + ), 1.60 min, UV detectable.

[0341] 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) [ka]

[0342] Step (i): To a solution of Intermediate 1, methyl 4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoate 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 h, then cooled to room temperature, filtered through a phase separator, and concentrated. The residue was dissolved in EtOAc, washed with water and brine, dried by passing through a hydrophobic frit, and concentrated. The residue was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 90% EtOAc 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 + ), 2.32 min, UV detectable.

[0343] Step (ii): A mixture of the 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 (3.7 mL) and water (0.9 mL) was heated to 100° C. in a microwave reactor for 30 min. The mixture was diluted with water and EtOAc, and the organic layer was separated, washed with water and brine, dried by passing through a hydrophobic frit, and concentrated. The residue was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 100% EtOAc 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 + ), 2.10 min, UV detectable.

[0344] 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) at −78° C. under a nitrogen atmosphere was added dropwise 1 M boron tribromide solution in DCM (6.08 mL, 6.08 mmol). The mixture was allowed to warm gradually to room temperature and stirred at room temperature for 18 hours. The mixture was cooled to 0° C. and MeOH / HO 1:1 (7.6 mL) was added dropwise. The mixture was stirred at room temperature for 1 hour, after which time 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. 1 H NMR(CD3OD,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), 5.11 (d, J = 7.1 Hz, 1H), 6.73-6.75 (m, 1H), 6.83-6.85 (m, 1H), 6.87-6.89 (m, 1H), 7.32 (d, J = 8.0 Hz, 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). No exchangeable protons were observed. (LC / MS Method D): m / z 554 [M+H] + (ES + ), 1.63 min, UV detectable.

[0345] 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) [ka]

[0346] Step (i): To a solution of Intermediate 1, methyl 4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoate 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 h, after which sodium triacetoxyborohydride (406 mg, 1.92 mmol) was added. The mixture was stirred at room temperature for 19 h, after which it was partitioned between 1 M aqueous NaOH and DCM. The organic layer was separated, and the aqueous layer was further extracted (3 times) with DCM. The organic phases were combined, washed with water and brine, dried by passing through a hydrophobic frit, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 80% EtOAc 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 + ), 2.59 min, UV detectable.

[0347] 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)boronic acid (72 mg, 0.48 mmol) were added, and the mixture was heated to 100 °C for an additional 30 min. The mixture was partitioned between water and EtOAc, and the organic layer was separated, washed with water and brine, dried by passing through a hydrophobic frit, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 90% EtOAc 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 + ), 2.45 min, UV detectable.

[0348] 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 and then concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 0.1% v / v acetic acid in isohexane with EtOAc (30% to 90%) 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 + ), 1.66 min, UV detectable.

[0349] 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). After stirring the mixture at room temperature for 18 h, additional sulfamoyl chloride (20 mg, 0.17 mmol) was added, and the mixture was stirred for an additional 18 h. Water (0.75 mL) was added, and the mixture was stirred for 30 min before being partitioned between EtOAc and 1 M aqueous HCl. The organic layer was separated, and the aqueous layer was further extracted with EtOAc (3 times). The combined organic layers were concentrated and triturated with 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 + ), 1.45 min, UV detectable.

[0350] 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 under a hydrogen atmosphere for 16 hours, 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. 1 H NMR (CD3OD, 400 MHz): δ (ppm) 1.51 (d, J = 7.0 Hz, 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 = 7.0 Hz, 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). No exchangeable protons were observed. (LC / MS Method D): m / z 570 [M+H] + (ES + ), 2.76 min, UV detectable.

[0351] Example 18: 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)-2-hydroxybenzoic acid (Compound 18) [ka]

[0352] Step (i): Intermediate 8, methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-carboxamido)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) under a nitrogen atmosphere. To this, 4 Å molecular sieves (0.5 g) and glacial acetic acid (4 μL, 0.065 mmol) were added at room temperature, and the mixture was stirred for 4 hours. After this time, the reaction mixture was cooled to 0 °C, and sodium cyanoborohydride (0.12 g, 1.95 mmol) was added in portions, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated, and the residue was partitioned between saturated aqueous NaHCO and EtOAc. The aqueous layer was separated and further extracted with EtOAc (3 times). The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) using a gradient of MeCN in water (0 to 44%) to give methyl 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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 + ), 1.37 min, UV detectable.

[0353] Step (ii): 4-((S)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)methyl 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 4 N aqueous HCl, adsorbed onto Celite, and purified by flash column chromatography (reverse-phase, C18) under 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-carboxamido)ethyl)-2-hydroxybenzoic acid (0.11 g, 0.21 mmol, 60% yield) as a white solid. 1 H NMR(DMSO,400MHz):δ(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). (LC / MS method E): m / z534[M+H] + (ES + ), 1.22 min, UV detectable.

[0354] Example 19: 4-((S)-1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)-2-hydroxybenzoic acid (Compound 19) [ka]

[0355] Step (i): To a solution of intermediate 8, methyl 2-hydroxy-4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate (500 mg, 1.62 mmol) and N,N-diisopropylethylamine (0.57 mL, 3.25 mmol) in MeCN (5 mL), 1-bromo-3-(bromomethyl)benzene (490 mg, 1.95 mmol) was added and the mixture was heated to 80° C. for 2 h. 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 organic layers were dried over Na2SO4 and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 0% to 34% EtOAc in hexanes to give methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-carboxamido)ethyl)-2-hydroxybenzoate (550 mg, 1.15 mmol, 71% yield) as a white solid. (LC / MS Method E): m / z 478 [M+H] + (ES + ), 1.94 min, UV detectable.

[0356] Step (ii): Methyl 4-((S)-1-((R)-4-(3-bromobenzyl)morpholine-3-carboxamido)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 1,4-dioxane:water mixture (1:1, 6 mL), and the mixture was purged with nitrogen gas at room temperature for 30 minutes. PdCl(dppf).DCM (43 mg, 0.05 mmol) was then added, and the reaction mixture was heated to 80 °C and maintained 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. The combined organic layers were dried over NaSO and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) using a gradient of MeCN (0 to 58%) in water to give methyl 4-((S)-1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)-2-hydroxybenzoate (170 mg, 0.32 mmol, 61%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), 1.52 min, UV detectable.

[0357] 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-carboxamido)ethyl)-2-hydroxybenzoate (170 mg, 0.32 mmol) in 1,4-dioxane (2 mL) and water (2 mL), lithium hydroxide monohydrate (73 mg, 1.60 mmol) was added and the mixture was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure, and the residue was acidified with 4 N HCl, directly adsorbed onto Celite, and purified by flash column chromatography (reverse-phase, C18) under 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-carboxamido)ethyl)-2-hydroxybenzoic acid (65 mg, 0.13 mmol, 39% yield) as an off-white solid. 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). (LC / MS method E): m / z518[M+H] + (ES + ), 1.35 min, UV detectable.

[0358] Example 20: 4-((S)-1-((R)-4-((2'-methyl-4'-(sulfamoyloxy)-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 20) [ka]

[0359] 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-carboxamido)ethyl)benzoic acid (65 mg, 0.13 mmol) in DMA (2 mL) at room temperature was added sulfamoyl chloride (157 mg, 1.37 mmol). The mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure. The residue was purified by flash column chromatography (reverse phase, C18) under 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-carboxamido)ethyl)benzoic acid (25 mg, 0.05 mmol, 33% yield) as an off-white solid. 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). (LC / MS Method E): m / z 554 [M+H] + (ES + ), 1.51 min, UV detectable.

[0360] Example 23: 4-[(1S)-1-[[(3R)-4-[[3-(4-carbamoylphenyl)-4-hydroxy-phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 23) [ka]

[0361] Step (i): A mixture of intermediate 1, methyl 4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate 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 h, after which sodium triacetoxyborohydride (812 mg, 3.83 mmol) was added. The mixture was stirred at room temperature for 16 h and then diluted with saturated aqueous NaHCO. The organic layer was separated, washed with water and brine, dried by passing through a hydrophobic frit, and concentrated. The residue was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 100% EtOAc 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 + ), 2.27 min, UV detectable.

[0362] 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 min. The mixture was partitioned between water and EtOAc, and the organic layer was separated, washed with water and brine, dried by passing through a hydrophobic frit, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of MeOH in DCM (0% to 10%) 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 pale orange solid. (LC / MS Method B): m / z 532 [M+H] + (ES + ), 2.00 min, UV detectable.

[0363] 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) at −78° C. under a nitrogen atmosphere was added dropwise a 1 M solution of boron tribromide in DCM (2.54 mL, 2.54 mmol). The mixture was allowed to warm gradually to room temperature and stirred at room temperature for 18 hours. The mixture was cooled to 0° C. and MeOH / HO 1:1 (3.4 mL) was added dropwise. The mixture was stirred at room temperature for 1 hour, after which time it was concentrated. The crude residue was purified by preparative HPLC [purification method A, 10→40% solvent B / solvent A] to give 4-[(1S)-1-[[(3R)-4-[[3-(4-carbamoylphenyl)-4-hydroxy-phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (54 mg, 0.11 mmol, 21% yield) as a white foam. 1 H NMR(CD3OD,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) , 5.08-5.16 (m, 1H), 6.93 (d, J = 8.3 Hz, 1H), 7.15 (dd, J = 8.3, 2.2 Hz, 1H), 7.30 (d, J = 2.2 Hz, 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). No exchangeable protons were observed. (LC / MS Method H): m / z 504 [M+H] + (ES + ), 1.64 min, UV detectable.

[0364] Example 26: 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)-2,6-difluorobenzoic acid, a single diastereomer of unknown stereochemistry (Compound 26) [ka]

[0365] Step (i): A mixture of (R)-methyl morpholine-3-carboxylate hydrochloride (220 mg, 1.21 mmol), Intermediate 11, 3'-(bromomethyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (360 mg, 1.19 mmol), and NaHCO (300 mg, 3.63 mmol) in MeCN was heated to 70 °C for 7 h. After this time, the mixture was concentrated under reduced pressure, and the residue was purified by flash column chromatography (normal phase, silica) using 10% MeOH / DCM to give (R)-methyl 4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxylate (420 mg, 1.14 mmol, 96% yield) as a yellow solid. (LC / MS Method I): m / z 369 [M+H] + (ES + ), 3.00 min, UV detectable.

[0366] Step (ii): To a solution of methyl (R)-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 h. 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 + ), 1.97 min, UV detectable.

[0367] 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, hydroxybenzotriazole (86 mg, 0.64 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (121 mg, 1.69 mmol) were added. The mixture was stirred at room temperature for 1 hour, and then triethylamine (171 mg, 1.69 mmol) and Intermediate 12, methyl 4-(1-aminoethyl)-2,6-difluorobenzoate (107 mg, 0.42 mmol) were added at 0 °C. The mixture was stirred at room temperature for 12 hours, then diluted with DCM, washed with 0.5 N HCl, dried over NaSO, and concentrated. The residue was purified by flash column chromatography (normal phase, silica) using 3% MeOH / DCM to give methyl 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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 + ), 3.10 min, UV detectable.

[0368] Step (iv): To a solution of methyl 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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, then concentrated and partitioned between EtoAc and water. The aqueous layer was isolated, adjusted to pH 4 with 1 N HCl, and extracted with EtOAc (3 times). The organic layers were combined, dried over Na2SO4, and concentrated. The crude material was purified by preparative HPLC [purification method F] to afford 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)-2,6-difluorobenzoic acid, a single diastereomer of unknown stereochemistry, 1 (50.0 mg, 0.09 mmol, 40% yield) as a white solid. 1 H NMR (CD3OD, 400 MHz): δ (ppm) 1.51 (d, J = 7.0 Hz, 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). No exchangeable protons were observed. (LC / MS Method I): m / z 538 [M+H] + (ES + ), 0.50 min, UV detectable.

[0369] Example 29: 4-((S)-1-((R)-4-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 29) [ka]

[0370] Step (i): To a solution of intermediate 3, methyl 4-((S)-1-((R)-morpholine-3-carboxamido)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) under a nitrogen atmosphere, 4 Å molecular sieves and glacial acetic acid (6 μL, 0.1 mmol) were added. The mixture was stirred at room temperature for 4 h, then 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 h and then partitioned between saturated aqueous NaHCO and EtOAc. The organic layer was separated, and the aqueous layer was further extracted with EtOAc. The combined organic layers were dried over NaSO and concentrated. The residue was purified by flash column chromatography (reverse phase, C18) using 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-carboxamido)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 boronic acid), 1.50 min, UV detectable.

[0371] Step (ii): Methyl 4-((S)-1-((R)-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine-3-carboxamido)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 gas at room temperature for 20 minutes, after which PdCl(dppf).DCM (14 mg, 0.026 mmol) was added, and the reaction mixture was heated to 80 °C for 2 hours. The mixture was cooled and partitioned between water and EtOAc. The organic layer was separated, and the aqueous layer was further extracted with EtOAc. The combined organic layers were dried over NaSO and concentrated, and the residue was purified by flash column chromatography (reverse phase, C18) under 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-carboxamido)ethyl)benzoate (45 mg, 0.081 mmol, 46% yield) as a white solid. (LC / MS Method E): m / z 554 [M+H] + (ES + ), 1.28 min, UV detectable.

[0372] Step (iii): To a solution of methyl 4-((S)-1-((R)-4-((2'-hydroxy-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoate (45 g, 0.08 mmol) in 1,4-dioxane (2 mL) and water (1 mL), lithium hydroxide monohydrate (17 mg, 0.40 mmol) was added 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) using 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-carboxamido)ethyl)benzoic acid (19 mg, 0.035 mmol, 44% yield) as a white solid. 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). (LC / MS method E): m / z540[M+H] + (ES + ), 1.44 min, UV detectable.

[0373] Example 30: 3-(4-carbamoyl-2-methylphenyl)-5-(((R)-3-(((S)-1-(4-carboxyphenyl)ethyl)carbamoyl)morpholino)methyl)pyridine 1-oxide (Compound 30) [ka]

[0374] Step (i): To a solution of 3-(4-carbamoyl-2-methylphenyl)-5-(chloromethyl)pyridine 1-oxide (200 mg, 0.72 mmol), Intermediate 14, in MeCN (12 mL) and DMF (2 mL) was added methyl 4-((S)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate hydrochloride (238 mg, 0.72 mmol), Intermediate 1, NaHCO (182 mg, 2.17 mmol), and sodium iodide (108 mg, 0.72 mmol). The mixture was heated to 70° C. for 4 h, then cooled, filtered, and concentrated under reduced pressure. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of MeOH (3% to 6%) in DCM to give 3-(4-carbamoyl-2-methylphenyl)-5-(((R)-3-(((S)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholino)methyl)pyridine 1-oxide (326 mg, 0.61 mmol, 85% yield) as a white solid. (LC / MS Method I): m / z 533 [M+H] + (ES + ), 2.73 min, UV detectable.

[0375] 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 (1.8 mL, 1.8 mmol) and the mixture was stirred at room temperature for 48 h. 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. 1H NMR (400 MHz, CD3OD) δ 1.51 (d, J = 7.0 Hz, 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 = 7.0 Hz, 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). No exchangeable protons were observed. (LC / MS Method I): m / z 519 [M+H] + (ES + ), 2.73 min, UV detectable.

[0376] Example 35: 4-((S)-1-((R)-4-((4'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 35) [ka]

[0377] Step (i): To a solution of Intermediate 1, methyl 4-[(1S)-1-[[(3R)-morpholine-3-carbonyl]amino]ethyl]benzoate hydrochloride (500 mg, 1.52 mmol) in MeCN (12 mL), potassium carbonate (525 mg, 3.8 mmol) and 3-bromobenzyl bromide (Intermediate 28, 760 mg, 3.0 mmol) were added. The reaction mixture was heated to 70° C. for 18 h, after which time the mixture was cooled and filtered through a phase separator. The solution was concentrated, and the residue was partitioned between EtoAc and water. The organic layer was separated, washed with brine, dried by passing through a hydrophobic frit, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 70% EtOAc 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 + ), 2.29 min, UV detectable.

[0378] Step (ii): A solution of methyl 4-[(1S)-1-[[(3R)-4-[(3-bromophenyl)methyl]morpholine-3-carbonyl]amino]ethyl]benzoate (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-hydroxybenzeneboronic 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 and held for 1 h. After this time, the mixture was diluted with EtOAc and water. The organic layer was separated, washed with brine, dried by passing through a hydrophobic frit, and concentrated. The crude material was purified by flash column chromatography (normal phase, silica) using a gradient of 30% to 100% EtOAc 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 + ), 2.15 min, UV detectable.

[0379] 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 h. The mixture was concentrated, and the crude residue was purified by reverse-phase preparative HPLC [Method B] to give 4-((S)-1-((R)-4-((4'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (25 mg, 0.054 mmol, 29% yield) as a gum, which was scratched to give an amorphous white solid. 1H NMR(CD3OD,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 = 12.8 Hz, 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). No exchangeable protons were observed. (LC / MS Method D): m / z 461 [M+H] + (ES + ), 2.23 min, UV detectable.

[0380] Example 36: (R)—N-((S)-1-(4-(1H-tetrazol-5-yl)phenyl)ethyl)-4-((4′-carbamoyl-5-hydroxy-2′-methyl-[1,1′-biphenyl]-3-yl)methyl)morpholine-3-carboxamide (Compound 36) [ka]

[0381] 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) at room temperature, 3'-(bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 6, 0.086 g, 0.27 mmol) was added, and the reaction mixture was stirred at 70 °C for 3 h. 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 dried over Na2SO4. The solvent was removed under reduced pressure and the crude product was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) in which the product was eluted with 0% to 55% acetonitrile / 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 + ), 1.54 min, UV detectable.

[0382] 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 h. After cooling to room temperature, the reaction mixture was 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 (Na2SO4). The solvent was removed under reduced pressure to give the crude product, which was purified by reverse-phase flash column chromatography (reverse-phase, C18 silica) in which the product was eluted with 0% to 60% acetonitrile / water to give (R)—N-((S)-1-(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.

[0383] 1 H 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). N of tetrazole H was not observed. (LC / MS Method E): m / z 542 [M+H]+ (ES + ), 1.52 min, UV detectable.

[0384] Example 37: 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 37) [ka]

[0385] Step (i): (S)-Morpholine-3-carboxylic acid hydrochloride (2.50 g, 14.97 mmol) was dissolved in acetone (15 mL) and HO (15 mL). KCO (10.40 g, 74.83 mmol) was added, and the reaction mixture was stirred at 0 °C for 30 min. BOC anhydride (5.22 g, 23.95 mmol) was added, and the reaction mixture was stirred at room temperature for 5 h. The reaction mixture was concentrated under reduced pressure, and the residue was then partitioned 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 (NaSO). The solvent was removed under reduced pressure 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 + ), 1.32 min, UV detectable.

[0386] 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)methyl benzoate (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 partitioned between water (250 mL) and EtOAc (100 mL), and the aqueous layer was further extracted with EtOAc (2 x 70 mL). The organic layers were combined and dried (Na2SO4). The crude material was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) in which the product was eluted with 0% to 70% acetonitrile / 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 + ), 1.82 min, UV detectable.

[0387] Step (iii): (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) under a nitrogen atmosphere. 4N HCl / dioxane (10 mL) was then added at room temperature, and the reaction mixture was stirred at room temperature for 4 hours. The solvent was removed under reduced pressure, and the crude material was purified by trituration with diethyl ether (15 mL) to give methyl 4-((S)-1-((S)-morpholine-3-carboxamido)ethyl)benzoate hydrochloride (2.60 g, 89.0%) as a white solid. (LC / MS Method E): m / z 293 [M+H] + (ES + ), 0.96 min, UV detectable.

[0388] Step (iv): Methyl 4-((S)-1-((S)-morpholine-3-carboxamido)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. Then, 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 under reduced pressure and the crude material was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) where the product was eluted with 0% to 70% acetonitrile / water to give methyl 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoate (0.25 g, 44.6%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), 1.31 min, UV detectable.

[0389] Step (v): Methyl 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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 for 4 hours. The reaction mixture was acidified with glacial acetic acid (1.5 mL), the pH was adjusted to ∼4, and the mixture was concentrated under reduced pressure. The crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) in which the product was eluted with 0% to 35% acetonitrile / water to give 4-((S)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (0.150 g, 73.4%) as a white solid.

[0390] 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=3.2HzandJ=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 = 7.4 Hz), 6.61 (s, 1H), 6.76 (d, 2H, J = 10.4 Hz), 7.23 (d, 1H, J = 8 Hz), 7.37 (t, 3H, J = 8.4 Hz), 7.73 (d, 1H, J = 8.0 Hz), 7.80 (d, 3H, J = 8.4 Hz), 7.96 (s, 1H), 8.50 (d, 1H, J = 8.4 Hz), 9.52 (s, 1H). No carboxylic acid protons were observed. (LC / MS Method E): m / z 518 [M+H] + (ES + ), 1.14 min, UV detectable.

[0391] Example 38: 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 38) [ka]

[0392] Step (i): (S)-Morpholine-3-carboxylic acid hydrochloride (2.50 g, 14.96 mmol) was dissolved in acetone (15 mL) and HO (15 mL). Potassium carbonate (10.40 g, 74.83 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. After this time, the reaction mixture was cooled to 0°C, and BOC anhydride (5.22 g, 23.95 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 5 hours and concentrated under reduced pressure. The residue was partitioned between water (100 mL) and DCM (50 mL), and the aqueous layer was further extracted with DCM (2 x 50 mL). The organic layers were combined and dried (NaSO). The solvent was removed under reduced pressure 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 + ), 1.32 min, UV detectable.

[0393] Step (ii): (S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (2.10 g, 9.09 mmol) was dissolved in DMF (20 mL), (R)-methyl 4-(1-aminoethyl)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 time, 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 partitioned between water (100 mL) and EtOAc (30 mL), and the aqueous layer was further extracted with EtOAc (2 x 50 mL). The organic layers were combined, dried (NaSO), and the resulting crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) in which the product was eluted with 0% to 60% acetonitrile / water to give tert-butyl (S)-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 + ), 1.74 min, UV detectable.

[0394] Step (iii): (S)-tert-Butyl 3-(((R)-1-(4-(methoxycarbonyl)phenyl)ethyl)carbamoyl)morpholine-4-carboxylate (3.00 g, 7.64 mmol) was dissolved in dioxane (10 mL) under a nitrogen atmosphere. Then, 4N HCl / dioxane (10 mL) was added at room temperature, and the reaction mixture was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure, and the crude product was purified by trituration with diethyl ether (15 mL) to give methyl 4-((R)-1-((S)-morpholine-3-carboxamido)ethyl)benzoate hydrochloride (2.50 g, 100%) as a white solid. (LC / MS Method E): m / z 293 [M+H] + (ES + ), 0.95 min, UV detectable.

[0395] Step (iv): Methyl 4-((R)-1-((S)-morpholine-3-carboxamido)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. Then, 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 under reduced pressure, and the crude material was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) in which the product was eluted with 0% to 70% acetonitrile / water to give methyl 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoate (0.35 g, 62.50%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), 1.23 min, UV detectable.

[0396] Step (v): Methyl 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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 approximately 4 and concentrated under reduced pressure. The crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) in which the product was eluted with 0% to 35% acetonitrile / water to give 4-((R)-1-((S)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (0.175 g, 51.5%) as a white solid.

[0397] 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=3.2HzandJ=9.2Hz),3.13(d,1H,J=12.8Hz),3.56-3.48(m,2H),3.76-3.69(m,3H),4.98(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,1 H,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). (LC / MS method E): m / z518[M+H] + (ES + ), 1.00 min, UV detectable.

[0398] Example 39: 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 39) [ka]

[0399] Step (i): (R)-morpholine-3-carboxylic acid hydrochloride (10.00 g, 59.87 mmol) was dissolved in acetone (50 mL) and HO (50 mL). Potassium carbonate (41.37 g, 299.31 mmol) was added, and the reaction mixture was stirred at 0 °C for 30 min. After this time, BOC anhydride (20.63 g, 89.79 mmol) was added, and the reaction mixture was stirred at room temperature for 6 h and concentrated under reduced pressure. The crude product was partitioned between water (500 mL) and EtOAc (200 mL), and the aqueous layer was further extracted with EtOAc (2 × 100 mL). The organic layers were combined and dried (NaSO). The solvent was removed under reduced pressure 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 + ), 1.50 min, UV detectable.

[0400] Step (ii): (R)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (0.5 g, 2.16 mmol) was dissolved in DMF (7 mL), and (R)-4-(1-aminoethyl)methyl benzoate (0.46 g, 2.59 mmol) was added to the reaction mixture at room temperature. HATU (1.23 g, 3.24 mmol) was then added, and the reaction mixture was stirred at room temperature for 30 minutes. N,N-diisopropylethylamine (1.12 mL, 6.49 mmol) was then added, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure and the crude product was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) where the product was eluted with 0% to 65% acetonitrile / 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 + ), 1.81 min, UV detectable.

[0401] Step (iii): (R)-tert-butyl 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 time, 4N HCl / dioxane (5 mL) was added at room temperature and the reaction mixture was stirred at room temperature for 4 hours. The solvent was removed under reduced pressure and the crude material was purified by trituration with diethyl ether (5 mL) to give 4-((R)-1-((R)-morpholine-3-carboxamido)ethyl)benzoate hydrochloride (0.4 g, 99.2%) as a white solid. (LC / MS Method E): m / z 293 [M+H] + (ES + ), 0.87 min, UV detectable.

[0402] Step (iv): Methyl 4-((R)-1-((R)-morpholine-3-carboxamido)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. Then, 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 under reduced pressure, and the crude product was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) in which the product was eluted with 0% to 70% acetonitrile / water to give methyl 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoate (0.2 g, 41.2%) as an off-white solid. (LC / MS Method E): m / z 532 [M+H] + (ES + ), 1.23 min, UV detectable.

[0403] Step (v): Methyl 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)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 approximately 4 and concentrated under reduced pressure. The crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) in which the product was eluted with 0% to 30% acetonitrile / water to give 4-((R)-1-((R)-4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (0.110 g, 56.5%) as a white solid.

[0404] 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=2.8HzandJ=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). (LC / MS method E): m / z518[M+H] + (ES + ), 1.14 min, UV detectable.

[0405] The following compounds are prepared according to the procedures described in the examples above, but using different intermediates.

[0406] Example 2: 4-[(1S)-1-[[(3R)-4-[[6-(4-carbamoyl-2-methyl-phenyl)-2-pyridyl]methyl]-morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 2)

[0407] Obtain the title compound according to the procedure described in Example 23, but using Intermediates 1, 2, and 15. Note: Step (ii): After adding additional catalyst, boronic ester, and base, microwave irradiation is continued for an additional hour.

[0408] HPLC Method A: Gradient 4% → 70% Solvent B / Solvent A. (LC / MS Method C): m / z 503 [M+H] + (ES + ), 1.74 min, UV detectable. 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).

[0409] Example 3: 4-[(1S)-1-[[(3R)-4-[[2-(4-carbamoyl-2-methyl-phenyl)-4-pyridyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 3)

[0410] The title compound is obtained according to the procedure described in Example 1, but using intermediates 1, 2, and 16. Note: Step (ii): After adding additional catalyst, boronic ester, and base, microwave irradiation is continued for an additional hour.

[0411] HPLC method B (LC / MS method C): m / z 503 [M+H] + (ES + ), 1.31 min, UV detectable. 1H 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).

[0412] Example 4: 4-[(1S)-1-[[(3R)-4-[[4-(4-carbamoyl-2-methyl-phenyl)-2-pyridyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 4)

[0413] Following the procedure described in Example 1, but using Intermediates 1, 2, 17, the title compound is obtained.

[0414] HPLC method B (LC / MS method C): m / z 503 [M+H] + (ES + ), 1.51 min, UV detectable. 1H 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).

[0415] Example 6: 6-[[[(3R)-4-[[3-(4-carbamoyl-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]methyl]pyridine-3-carboxylic acid (Compound 6)

[0416] Following the procedure described in Example 1, but using intermediates 2, 18, and 19, the title compound is obtained.

[0417] HPLC method C (LC / MS method C): m / z 487 [MH] - (ES - ), 1.79 min, UV detectable. 1H NMR(CD3OD,400MHz):δ(ppm)2.25-2.32(m,4H),2.76-2.85(m,1H),3.12(dd,J=9.5,3.7Hz,1 H),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).

[0418] Example 7: 4-((S)-1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 7)

[0419] Following the procedure described in Example 5, but using intermediates 2 and 3, the title compound is obtained.

[0420] The product is purified by flash chromatography (reverse phase). (LC / MS Method E): m / z 502 [M+H] + (ES + ), 1.42 min, UV detectable. 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).

[0421] 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)

[0422] The title compound is obtained according to the procedure described in Example 1, but using intermediates 2, 19, and 20. Note: Step (ii): Additional catalyst, boronic ester, and base are added, followed by microwave irradiation for an additional 2.5 hours.

[0423] HPLC method D (LC / MS method C): m / z524[M+H] + (ES + ), 1.83 min, UV detectable. 1 H NMR(CD3OD,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).

[0424] Example 12: 4-[(1S)-1-[[(3R)-4-[[5-(2-methyl-4-sulfamoyl-phenyl)-3-pyridyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 12)

[0425] Following the procedure described in Example 1, but using intermediates 1, 4, and 21, the title compound is obtained.

[0426] HPLC method D (LC / MS Method D): m / z 539 [M+H] + (ES +), 1.78 min, UV detectable. 1 H NMR(CD3OD,400MHz):δ(ppm)1.49(d,J=7.0Hz,3H),2.31(s,3H),2.90-3.02(m,1H),3.1 1-3.19(m,1H),3.62-3.75(m,2H),3.78(dd,J=10.1,3.3Hz,1H),3.91-3.98(m,2H),4.1 0-4.19(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).

[0427] Example 13: 4-[[[(3R)-4-[[3-(4-carbamoyl-2-methyl-phenyl)phenyl]methyl]morpholine-3-carbonyl]amino]methyl]-2-fluoro-benzoic acid (Compound 13)

[0428] Following the procedure described in Example 1, but using intermediates 2, 19, and 22, the title compound is obtained.

[0429] HPLC method C (LC / MS method D): m / z 506 [M + H] + (ES + ), 2.31 min, UV detectable. 1 H NMR(CD3OD,400MHz):δ(ppm)2.22-2.32(m,4H),2.76-2.83(m,1H),3.08(dd,J=9.5,3.6Hz,1H ),3.24-3.31(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. 29-7.34(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).

[0430] Example 14: 4-((S)-1-((R)-4-((4'-carbamoyl-2'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 14)

[0431] Following the procedure described in Example 5, but using intermediates 3 and 24, the title compound is obtained. Note: Step (iii): The compound is sufficiently pure after work-up; no chromatography is required.

[0432] The product is purified by acid / base extraction. (LC / MS Method E): m / z 504 [M+H] + (ES + ), 1.16 min, UV detectable. 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).

[0433] Example 16: 4-[(1S)-1-[[(3R)-4-[[2-(2-methyl-4-sulfamoyl-phenyl)-4-pyridyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 16)

[0434] The title compound is obtained according to the procedure described in Example 1, but using intermediates 1, 4, and 16. Note: Step (ii): After the addition of additional catalyst and boronic ester, microwave irradiation is continued for an additional hour.

[0435] HPLC method D (LC / MS Method D): m / z 539 [M+H] + (ES + ), 1.98 min, UV detectable. 1 H 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).

[0436] Example 17: 4-[(1S)-1-[[(3R)-4-[[4-(2-methyl-4-sulfamoyl-phenyl)-2-pyridyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 17)

[0437] Following the procedure described in Example 1, but using intermediates 1, 4, and 17, the title compound is obtained.

[0438] HPLC method D (LC / MS Method D): m / z 539 [M+H] + (ES + ), 1.73 min, UV detectable. 1H 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).

[0439] Example 21: 2-hydroxy-4-((S)-1-((R)-4-((5-(2-methyl-4-sulfamoylphenyl)pyridin-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 21)

[0440] Following the procedure described in Example 18, but using intermediates 8 and 26, the title compound is obtained.

[0441] The product is purified by flash chromatography (reverse phase). (LC / MS Method E): m / z 555 [M+H] + (ES + ), 1.26 min, UV detectable. 1H 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).

[0442] Example 22: 2-hydroxy-4-((S)-1-((R)-4-((5-hydroxy-2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 22)

[0443] Following the procedure described in Example 18, but using intermediates 8 and 27, the title compound is obtained.

[0444] The product is purified by flash chromatography (reverse phase). (LC / MS Method E): m / z 570 [M+H] + (ES + ), 1.29 min, UV detectable. 1 H 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).

[0445] Example 24: 4-[(1S)-1-[[(3R)-4-[[4-hydroxy-3-(4-sulfamoylphenyl)phenyl]methyl]morpholine-3-carbonyl]amino]ethyl]benzoic acid (Compound 24)

[0446] Following the procedure described in Example 26, but using intermediates 1 and 29, the title compound is obtained.

[0447] HPLC method D (LC / MS method H): m / z540[M+H] + (ES + ), 1.76 min, UV detectable. 1 H NMR(CD3OD,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).

[0448] Example 25: 2-hydroxy-4-((S)-1-((R)-4-((2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)benzoic acid (Compound 25)

[0449] Following the procedure described in Example 19, but using intermediates 8 and 4, the title compound is obtained. Note: Step (ii): 1.2 equivalents of the boronic ester are used. Step (iii): The reaction mixture is partitioned between EtOAc and 4N HCl; the organic layer is isolated and concentrated, and the residue is purified by preparative HPLC.

[0450] HPLC method G (LC / MS Method E): m / z 554 [M+H] + (ES + ), 1.68 min, UV detectable. 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),7.5 9(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).

[0451] Example 27: 4-(1-((R)-4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)ethyl)-2,6-difluorobenzoic acid (single diastereomer 2 of unknown stereochemistry, Compound 27)

[0452] Following the procedure described in Example 29, but using intermediates 11 and 30, the title compound is obtained.

[0453] HPLC method F (LC / MS Method I): m / z 538 [M+H] + (ES + ), 0.57 min, UV detectable. 1H NMR(CD3OD,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).

[0454] Example 28: 4-((S)-1-((R)-4-((5-(4-carbamoyl-2-methylphenyl)pyridin-3-yl)methyl)morpholine-3-carboxamido)ethyl)-2-hydroxybenzoic acid (Compound 28)

[0455] Following the procedure described in Example 18, but using intermediates 8 and 31, the title compound is obtained.

[0456] HPLC method E (LC / MS Method E): m / z 519 [M+H] + (ES + ), 1.36 min, UV detectable. 1H 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).

[0457] Example 31: (R)-4-(1-(4-((4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)cyclopropyl)benzoic acid (Compound 31)

[0458] Following the procedure described in Example 19, but using intermediates 2 and 32, the title compound is obtained. Note: Step (i): 3 equivalents of base are used. Step (ii): 1.5 equivalents of boronic ester are used.

[0459] The product is purified by flash chromatography (reverse phase). (LC / MS Method E): m / z 514 [M+H] + (ES + ), 1.46 min, UV detectable. 1H 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).

[0460] Example 32: (R)-4-(1-(4-((4'-carbamoyl-2'-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)cyclopropyl)benzoic acid (Compound 32)

[0461] Following the procedure described in Example 19, but using intermediates 24 and 32, the title compound is obtained. Note: Step (i): 3 equivalents of base are used. Step (ii): 1.5 equivalents of boronic ester are used.

[0462] The product is purified by flash chromatography (reverse phase). (LC / MS Method E): m / z 516 [M+H] + (ES + ), 1.45 min, UV detectable. 1 H 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).

[0463] Example 33: (R)-4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)cyclopropyl)benzoic acid (Compound 33) [ka]

[0464] Step (i) (R)-4-(1-(morpholine-3-carboxamido)cyclopropyl)methyl benzoate, HCl salt (Intermediate 32, 0.25 g, 0.75 mmol) and DIPEA (0.4 mL, 2.26 mmol) were dissolved in acetonitrile (3 mL) at room temperature. 3'-(Bromomethyl)-5'-hydroxy-2-methyl-[1,1'-biphenyl]-4-carboxamide (Intermediate 6, 0.24 g, 0.75 mmol) was added, and the reaction mixture was stirred at 70 °C for 2 h. The reaction mixture was then partitioned between water (70 mL) and EtOAc (70 mL). The aqueous layer was further extracted with EtOAc (2 x 25 mL), and the organic layers were combined and dried (NaSO). The solvent was removed under reduced pressure and the crude material was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) in which the product was eluted with 0% to 56% acetonitrile / water to give pure (R)-4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)cyclopropyl)methyl benzoate (0.20 g, 49%) as a white solid. (LC / MS Method E): m / z 544 [M+H] + (ES + ), 1.56 min, UV detectable.

[0465] Step (ii) (R)-4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)cyclopropyl)methyl 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 about pH 3 with glacial acetic acid (0.3 mL), and the reaction mixture was concentrated under reduced pressure. The crude product was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) in which the product was eluted with 0% to 22% acetonitrile / water to give pure (R)-4-(1-(4-((4'-carbamoyl-5-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)cyclopropyl)benzoic acid (0.13 g, 66.7%) as an off-white solid. (LC / MS Method E): m / z 530 [M+H] + (ES + ), 1.31 min, UV detectable. 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).

[0466] Example 34: (R)-4-(1-(4-((4'-carbamoyl-6-hydroxy-[1,1'-biphenyl]-3-yl)methyl)morpholine-3-carboxamido)cyclopropyl)benzoic acid (Compound 34)

[0467] Following the procedure described in Example 5, but using intermediates 33 and 34, the title compound is obtained. Note: Steps (ii) and (iii) only.

[0468] The product is purified by flash chromatography (reverse phase). (LC / MS Method E): m / z 516 [M+H] + (ES + ), 1.07 min, UV detectable. 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). [Table 2-1] [Table 2-2] [Table 2-3]

[0469] biological activity Cloning, baculovirus production, large-scale infection of HEK293 cells, and membrane preparation: Human prostaglandin E2 receptor 4 (EP4) was cloned into the pBacMam expression vector (GenScript, UK). EP4 DNA transfer was performed using the Invitrogen Bac-to-Bac Baculovirus Expression Systems. P0 baculovirus was generated by transfecting bacmid DNA into SF9 cells using Cellfectin II transfection reagent (Thermo Fisher Scientific, UK). Following the P0 generation, P1 virus was generated and prepared for large-scale infection and membrane preparation. HEK293 cells were grown in DMEM (Thermo Fisher Scientific, UK) supplemented with 10% heat-inactivated fetal bovine serum (FBS). Cells were cultured at 3.5 × 10 6 Seeding density of cells / mL, 500cm 3 The cells were infected with 5% v / v EP4BacMan in a flask. Expression was carried out for 36 hours at 37°C and 5% CO2. Cells were detached using PBS and a cell scraper. The cell culture was centrifuged at 2500 rpm at 4°C for 10 minutes. The supernatant was then decanted, and the pellet was stored at -80°C. The pellet was thawed and resuspended in 15 mL of homogenization buffer (20 mM HEPES, 10 mM EDTA, pH 7.4). It was then homogenized for 10 seconds using a mechanical homogenizer (VMR). The membranes were centrifuged in a centrifuge tube at 40,000g for 15 minutes at 4°C. The supernatant was decanted and resuspended in 15 mL of homogenization buffer. The membranes were homogenized for 20 seconds. The membranes were centrifuged at 40,000g for 45 minutes at 4°C. The membranes were resuspended in 3 mL of storage buffer (20 mM HEPES, 0.1 mM EDTA, pH 7.4) and mixed well, after which they were stored at -80°C.

[0470] cAMP Gs functional assay: cAMP production after EP4 receptor activation was measured using the Homogeneous Time-Resolved Fluorescence (HTRF) cAMP Dynamic-2 Assay (Cisbio, France). HEK293 cells were transfected with 0.5% EP4 Bacmam virus for 36 hours, then detached and frozen at -150°C.

[0471] On the day of the test, increasing concentrations of test compound were added to a ProxiPlate-384 Plus, White 384-shallow well Microplate (PerkinElmer, USA) using ECHO dispensing, alongside a positive control (10 μM PGE2 (Tocris, Abingdon, UK)) and a negative control (DMSO (Sigma-Aldrich, UK)).

[0472] Cells were thawed in a water bath, resuspended in DMEM supplemented with 10% FBS, and then pelleted by centrifugation at 1200 rpm for 5 minutes. The pellet was resuspended in assay buffer (DMEM + 0.5 mM IBMX (Tocris, Abingdon, UK)) at 0.5x10 6 The cells were resuspended at 1000 cells / mL. The cell suspension was added to the dispensed assay plate using a Multidrop at a final assay concentration of 5000 cells / well. The plate was then incubated at 37°C, 5% CO for 30 minutes. cAMP production was measured according to the manufacturer's instructions, and the plate was read on a PheraStar fluorescent plate reader (BMG Labtec, Germany).

[0473] As shown in Table 3, pEC 50 Values (-Log of molarity (M) in moles / liter) were calculated from the midpoint of the curve using Dotmatics.

[0474] [Table 3]

[0475] Unidirectional CACO-2 cell permeability assay Caco-2 cells (ECACC) were plated at 2x10 5 Cells were seeded at 1000 x g / well and cultured at 37°C under 5% CO2 for 21 days before use as a confluent monolayer. Each test compound was incubated in duplicate with a final concentration of 10 μM in assay buffer (Hank's Balanced Salt Solution, pH 6.5, supplemented with 25 mM HEPES). The basolateral chamber (serving as a receiver) was supplemented with 25 mM HEPES and adjusted to pH 7.4 (final concentration: 0.2% DMSO).

[0476] Incubations were carried out at 37°C, and samples were removed from both the donor and acceptor chambers at T = 0 and 1 h, and the compounds were analyzed by mass spectrometry (LC-MS / MS), including an analytical internal standard (0.5 μM carbamazepine).

[0477] Apparent transmittance (P app ) values are shown in Table 4 and are determined from the following relationship:

[0478]

number

[0479] where V is the volume of each compartment of the Transwell (125 μL apical, 600 μL basolateral), each concentration is the relative MS response value (normalized to an internal standard) of the compound in the donor chamber before incubation and in the acceptor chamber at the end of incubation, and area = the area of the cell exposed to drug translocation (0.33 cm). 2 ).

[0480] Lucifer Yellow (LY) was added to the apical buffer in all wells to assess cell layer viability. Because LY cannot freely penetrate lipophilic barriers, a high degree of LY transport indicates poor cell layer integrity, and the P app is 10x10 -6 Wells exceeding cm / sec were excluded.

[0481] The recovery rate of compound from each well was determined from the MS response values (normalized to an internal standard) in the donor and acceptor chambers at the end of incubation, compared with the response value in the donor chamber before incubation.

[0482] [Table 4]

[0483] While the present invention has been described in connection with its detailed description, it should be understood that the above description is intended to be illustrative and not limiting of the scope of the invention, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. Numbered Embodiments 1. Compounds of Formula I: [ka] or a pharmaceutically acceptable salt or tautomer thereof; 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 of each other, CR″ or N, and at least one of X and Y is CH; R 1 and R 2are, independently of each other, H, C 1-6 Alkyl, C 1-6 Alkoxy or R 1 and R 2 together with the carbon atoms to which they are attached, form C 3-6 forming a cycloalkane-1,1-diyl ring; Each R 3 are, independently of each other, H, OR', COOR', C(O)R', halo, and C 1-6 alkyl; R 4 is H, C 1-6 alkyl, halo, CN, NO2, OR', CO2R', or C(O)R'; R 5 is OR', OC(O)R', OC(O)OR', COR', CON(R')2, SON(R')2, SOR', OSOR', or OSON(R')2; Each R' is independently H, C 1-6 Alkyl, or C 3-6 is cycloalkyl; Each R" is H, C 1-6 alkyl, halo, or OR'; n is 0, 1, 2, or 3; Each occurrence of alkyl and cycloalkyl may optionally, and independently of one another, be selected from up to three of OH, SH, CN, NO, COH, halo, or COOC. 1-4 is substituted with alkyl; Heterocycloalkyl, aryl, and heteroaryl each optionally, and independently of one another, may be selected from up to three of OR', SR', CN, NO2, CO2R', halo, C 1-4 substituted with alkyl or oxo; A compound of formula I, or a pharmaceutically acceptable salt or tautomer thereof. 2. Ring B is a 5- to 6-membered aryl or a 5- to 6-membered heteroaryl, each of which optionally, and independently of each other, contains up to three of OR', SR', CN, NO2, CO2R', halo, C 1-4The compound of embodiment 1, which is substituted with alkyl, or oxo. 3. The compound of embodiment 1, wherein Ring B is phenyl optionally substituted with up to three OH, or a 6-membered heteroaryl containing one or two nitrogen atoms, each of which is optionally oxidized. 4. A compound of formula (1), [ka] or a pharmaceutically acceptable salt or tautomer thereof; A is, [ka] selected from the group consisting of: U, V, W, and Z are each independently CH, COH, N, or N + -O - wherein at least three of U, V, W, and Z are CH; X and Y are each, independently of one another, selected from the group consisting of CH, CF, COH, or N; R 1 and R 2 are each independently H, C optionally substituted with 1 to 3 fluorine atoms, 1-3 alkyl or R 2 C optionally substituted with 1 to 3 fluorine atoms by bonding to 3-6 forming a cycloalkyl ring; R 3 is H, OH, or F; R 4 is H, OH, CN, halo, C optionally substituted with 1 to 3 fluorine atoms 1-3 Alkoxy or C optionally substituted with 1 to 3 fluorine atoms 1-3 is alkyl; R 5 is OH, CO2H, CONH2, SO2NH2, or OSO2NH2; R 6 is C1-3 Alkyl or C 3-6 is a cycloalkyl ring, A compound of formula I, or a pharmaceutically acceptable salt or tautomer thereof. 5. A, [ka] 5. The compound of embodiment 4, wherein 6. Compounds of formula (2a) or (2b): [ka] or a pharmaceutically acceptable salt thereof, U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , and R 5 is the same as defined in embodiment 4. The compound of embodiment 4. 7. R 1 is H or methyl, or R 2 to form a cyclopropane-1,1-diyl ring. 8. R 1 is methyl. 9. R 2 The compound of any one of embodiments 1 to 6, wherein is H. 10. Compounds of formula (3a) or (3b): [ka] or a pharmaceutically acceptable salt thereof, U, V, W, X, Y, Z, R 1 , R 2 , R 3 , R 4 , and R 5 is the same as defined in embodiment 4. The compound of embodiment 4. 11. R 3 The compound of any one of embodiments 1 to 10, wherein is H. 12. R 4 The compound of any one of embodiments 1 to 11, wherein is H, OH, or methyl. 13. R 4 The compound of embodiment 12, wherein is methyl. 14. R 5 The compound of any one of embodiments 1 to 13, wherein is CONH2 or SO2NH2. 15. The compound of any one of embodiments 4 to 14, wherein X and Y are both CH. 16. The compound of any one of embodiments 4 to 15, wherein U, V, W, and Z are CH. 17. The compound of any one of embodiments 4 to 15, wherein U, V, and Z are CH and W is COH. 18. [ka] [ka] [ka] or a pharmaceutically acceptable salt thereof. 19. 19. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 18 and a pharmaceutically acceptable excipient. 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. twenty one. The compound or composition for use according to embodiment 20, wherein the EP4 receptor mediated disorder is a gastrointestinal disorder. twenty two. 22. The compound or composition for use according to embodiment 21, wherein the gastrointestinal disorder is selected from the group consisting of constipation, 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 dysfunction, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced enteropathy, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrhea, immune-mediated gastrointestinal disorders, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis. twenty three. The compound or composition for use according to embodiment 20, wherein said EP4 receptor mediated disease is a pulmonary disease or condition. twenty four. 24. The compound or composition for use according to embodiment 23, wherein the pulmonary disease or condition is selected from chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, reactive airways disease, and idiopathic pulmonary fibrosis.

Claims

1. Compounds of Formula I: 【Chemical 1】 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; 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; Ring B is aryl or heteroaryl; X and Y are each, independently of one another, CR″ or N, and at least one of X and Y is CH; R １ and R ２ are each independently H, C １－６ Alkyl, C １－６ Alkoxy or R １ and R ２ together with the carbon atoms to which they are attached, form C ３－６ forming a cycloalkane-1,1-diyl ring; Each R ３ are, independently of each other, H, OR', COOR', C(O)R', halo, and C １－６ alkyl; R ４ is H, C １－６ alkyl, C optionally substituted with 1 to 3 fluorine atoms １－３ Alkoxy, halo, CN, NO ２ , OR', CO ２ R′, or C(O)R′; R ５ is OR', OC(O)R', OC(O)OR', CO ２ R', CON(R') ２ , S.O. ２ N(R') ２ , S.O. ２ R', OSO ２ R', or OSO ２ N(R') ２ and Each R' is independently H, C, or １－６ Alkyl, or C ３－６ is cycloalkyl; Each R" is H, C １－６ alkyl, halo, or OR'; n is 0, 1, 2, or 3; Each occurrence of alkyl and cycloalkyl may optionally, and independently of one another, be selected from the group consisting of up to three OH, SH, CN, NO, ２ , CO ２ H, halo, or COOC １－４ is substituted with alkyl; Heterocycloalkyl, aryl, and heteroaryl each optionally and independently of one another may be selected from up to three groups: OR', SR', CN, NO. ２ , CO ２ R', Halo, C １－４ substituted with alkyl or oxo; A compound of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

2. Ring B is a 5- to 6-membered aryl or a 5- to 6-membered heteroaryl, each of which optionally and independently of each other contains up to three of OR', SR', CN, NO ２ , CO ２ R', Halo, C １－４ The compound of claim 1 , which is substituted with alkyl or oxo.

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

4. 4. A compound according to any one of claims 1 to 3, wherein n is 0, 1 or 2, preferably n is 0 or 1.

5. Compound of formula (1): 【Chemistry 2】 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; U, V, W, and Z are each independently CH, COH, N, or N ＋ -O － wherein at least three of U, V, W, and Z are CH; A compound according to any one of claims 1 to 4.

6. A is, 【Chemistry 3】 selected from the group consisting of: R ６ is C １－３ Alkyl or C ３－６ is a cycloalkyl ring; Preferably, A is 【Chemistry 4】 and more preferably, A is 【Chemistry 5】 That is, A compound according to any one of claims 1 to 5.

7. A compound of formula (2a) or (2b), 【Chemistry 6】 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; U, V, W, X, Y, Z, R １ , R ２ , R ３ , R ４ , and R ５ is the same as defined in claim 5, The compound of claim 5.

8. R １ and R ２ are each independently selected from H, C optionally substituted with 1 to 3 fluorine atoms, １－３ alkyl, or R １ is R ２ C optionally substituted with 1 to 3 fluorine atoms by being bonded to ３－６ 8. The compound according to any one of claims 1 to 7, which forms a cycloalkyl ring.

9. R １ is H or methyl, or R １ is R ２ to form a cyclopropane-1,1-diyl ring; preferably, R １ 9. The compound of claim 1, wherein is methyl.

10. R ２ 10. The compound of claim 1, wherein is H.

11. A compound of formula (3a) or (3b), 【Chemistry 7】 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof; U, V, W, X, Y, Z, R ３ , R ４ , and R ５ is the same as defined in claim 5, The compound of claim 5.

12. R ３ is selected from the group consisting of H, OH, or F; preferably, R ３ is H or F; more preferably, R ３ 12. The compound of claim 1, wherein is H.

13. R ４ is H, OH, CN, halo, optionally substituted with 1 to 3 fluorine atoms １－３ Alkoxy or C optionally substituted with 1 to 3 fluorine atoms １－３ alkyl; preferably, R ４ is H, OH, or methyl; more preferably, R ４ 13. The compound of claim 1, wherein is methyl.

14. R ５ OH, CO ２ H, CONH ２ , S.O. ２ NH ２ , or OSO ２ NH ２ Preferably, R ５ OH, CONH ２ , S.O. ２ NH ２ , or OSO ２ NH ２ and more preferably, R ５ Gonna be CONH ２ or SO ２ NH ２ 14. The compound of any one of claims 1 to 13, wherein

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

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

17. 17. The compound of any one of claims 5 to 16, wherein U, V, W, and Z are CH, or U, V, and Z are CH and W is COH. 【Request 18】 【Chemical 8-1】 【Chemistry 8-2】 【Chemistry 8-3】 2. The compound of claim 1 selected from: or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

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

20. 20. The pharmaceutical composition of claim 19, further comprising at least one additional therapeutic agent selected from the group consisting of an aminosalicylates, corticosteroids, immunomodulators, and combinations thereof.

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

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

25. 25. The compound, composition, or kit for use according to claim 24, wherein the gastrointestinal disorder is selected from the group consisting of constipation, 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 dysfunction, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorder, functional gastrointestinal disorder, drug-induced bowel disease, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux, diarrhea, immune-mediated gastrointestinal disease, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis.

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

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

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