1,2-substituted 3-oxopyrazolidine 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 I as prostaglandin E2 receptor 4 (EP4) agonists for use in methods for treating gastrointestinal or pulmonary diseases or disorders. An exemplary compound is, for example, 4-(2-(2-(3-hydroxy-3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (Example 1). For example, pharmacological data are provided. JPEG2025527174000052.jpg57170

Description

[Technical Field]

[0001] This application relates to novel compounds and their uses as prostaglandin E2 receptor 4 (EP4) agonists. The compounds described herein may be useful for treating or preventing diseases in which the EP4 receptor is involved. This application also relates to pharmaceutical compositions containing these compounds, as well as the preparation 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 metabolic derivatives of arachidonic acid that play important roles in cellular physiology. Arachidonic acid is an endogenous component of membrane phospholipids, released by the activity of phospholipase A2 (PLA2). Prostaglandin biosynthesis is mediated by cyclooxygenase (COX), which catalyzes the conversion of arachidonic acid to an unstable intermediate (PGH2), resulting in the production of prostaglandins, including PGE2. PGE2 is the most widely produced prostanoid, and its activity is mediated by the action of 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 the signaling pathway: i) EP2 and EP4, which are Gs-coupled receptors (activating adenylate cyclase and generating cyclic adenosine monophosphate (cAMP)), ii) EP1, which is a Gq-coupled receptor (activating PLC), and iii) EP3, which is a Gi-coupled receptor (inhibiting adenylate cyclase).

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

[0005] The EP4 receptor plays an important role in diverse 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 to treat 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%, negatively impacting quality of life (QoL) and resulting in significant societal and economic burdens. Chronic constipation can cause discomfort to patients and affect their daily lives. Symptoms include hard, lumpy stools, straining during defecation, and a feeling of incomplete evacuation. The use of laxatives and stool softeners remains high, despite many patients not deriving substantial benefit. There remains a need for treatments that can provide complete and sustained symptom relief.

[0007] The intestinal mucosa plays a crucial role in maintaining anion and fluid homeostasis. These functions are mediated by coordinated ion transport through membrane-bound transporters and channels located on the apical and basolateral membranes of intestinal epithelial cells. PGE2 is a well-established secretagogue that can directly promote chloride secretion from intestinal epithelial cells. The secretory effect 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 have therapeutic value in promoting intestinal fluid homeostasis in chronic constipation conditions.

[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 includes strategies to control abnormal, dysregulated immune responses in the intestinal mucosa. Depending on the severity of the disease, a wide variety of medications are used to induce and maintain remission. These include aminosalicylates, corticosteroids, immunosuppressants, antibiotics, and biologics. This can be administered in a stepwise approach, with therapy intensifying 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. Current treatment strategies achieve long-term remission in only a small proportion of patients, suggesting the need for improved therapy.

[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 many cell types, including gastrointestinal (GI) epithelial cells, lamina propria mononuclear cells, and colonic innervating sensory neurons, and may be beneficial in attenuating inappropriate mucosal immune responses and protecting the GI mucosal barrier. 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 EP4 agonists provides benefit 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 mutant mice lacking EP4 develop severe dextran sulfate sodium-induced colitis characterized by impaired mucosal barrier function, increased epithelial cell loss, crypt damage, and increased immune cell infiltration (Kabashima, K. et al. J. Clin. Invest. 2002, 109, 883-893). 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 efficacy was not evaluated in this study, patients treated with ONO-4819CD showed signs of improvement in Disease Activity Index (DAI) 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 airflow limitation. Approximately 300 million people are estimated to 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 can lead to mortality and morbidity. There remains a need for therapies that can effectively control asthma symptoms and reduce the risk of future exacerbations for 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 caliber in patients with chronic bronchitis and asthma. However, PGE2 also induces reflex coughing, likely via EP3 receptor-mediated upper airway stimulation. This has led to efforts to discover EP receptor-selective drugs for the treatment of airway disorders. Interestingly, significant species differences have been reported in the receptor subtypes responsible for mediating airway smooth muscle relaxation. In guinea pigs, monkeys, and mice, EP2 agonists can induce relaxation of airway smooth muscle, but in humans, this is mediated through EP4 receptors (Buckley, J. et al. Thorax 2011, 66, 1029-1035).Selective EP4 receptor agonists may have potential therapeutic value in airway diseases.

[0011] The EP4 receptor has been shown to play a role in regulating blood pressure. EP4 is expressed on smooth muscle and endothelial cells and can induce vasodilatory effects by endothelial nitric oxide synthase (eNOS)-mediated nitric oxide (NO) production. PGE2 has been shown to dose-dependently relax aortic smooth muscle, an effect that is abolished in EP4 knockout mice. In halothane-anesthetized dogs, the EP4-selective agonist ONO-AE1-329 induced vasoconstriction (Honda, A. et al. Eur. J. Pharmacol. 2016, 775, 130-137). Similarly, hypotension has been reported as an adverse drug reaction in IBD patients receiving ONO-4819CD (Nakase, H. et al. Inflamm. Bowel Dis. 2010, 16, 731-733). EP4 agonists that can be used to treat various gastrointestinal and respiratory tract diseases without cardiovascular side effects may have potential therapeutic value.In particular, EP4 agonists that can be used to treat various gastrointestinal disorders without cardiovascular side effects may have potential therapeutic value.Discovery of safe and effective EP4 selective agents is needed. Summary of the Invention

[0012] The present invention provides compounds that have activity as prostaglandin E2 receptor 4 (EP4) agonists.

[0013] In one aspect, provided herein are compounds of formula I: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof [In the formula, A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R 3 , S(O)2R', S(O)2OR', SO2N(R')2, C 1~8 alkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; X is halo, OR', COOR', or C1~6 is alkyl, L and L' are each independently C 2~4 is alkylene, R 1 H, Halo, CN, NO2, OR', SR', COOR', C 1~6 Alkoxy or C 1~6 is alkyl, R 2 , OR', OC(O)R 3 ,OC(O)OR 3 , CO2R', CON(R')2, SO2N(R')2, SO2R 3 , OSO2R 3 or OSO2N(R')2, R 3 is C 1~6 Alkyl, C 3~6 cycloalkyl, or phenyl; R' is H, C 1~6 Alkyl or C 3~6 is cycloalkyl, n is 0, 1, 2, or 3; In each instance, alkyl, alkylene, alkoxy, and cycloalkyl are each optionally and independently selected from OH, SH, CN, NO, COOH, halo, or COOC. 1~4 substituted with up to three of alkyl, In each instance, heterocycloalkyl, aryl, and heteroaryl are each optionally and independently selected from OR′, SR′, CN, NO 2 , CO 2 R′, halo, C 1~4 substituted with up to three of the following: alkyl, or oxo is provided.

[0014] In another aspect, the invention includes pharmaceutical compositions comprising a compound described herein and a pharmaceutically acceptable excipient.

[0015] In another aspect, the invention includes a kit comprising a compound described herein and at least one additional therapeutic agent selected from the group consisting of an aminosalicylate, a corticosteroid, an immunomodulator, and combinations thereof.

[0016] In another aspect, the invention includes a compound described herein, a composition described herein, or a kit described herein for use as a medicament.

[0017] 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.

[0018] 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.

[0019] In another aspect, the present invention includes a method for treating an EP4 receptor-mediated disease, comprising administering to a patient in need thereof a compound or composition described herein.

[0020] The compounds of the present invention can be used as EP4 receptor agonists.The compounds of the present invention can selectively act on EP4 receptors.These compounds can be used to manufacture compositions or medicaments.The compounds, compositions or medicaments can be used to treat, prevent, improve, control, or reduce the risk of diseases or disorders associated with EP4 receptors. The compound, composition, or medicament may be used to treat, prevent, ameliorate, control, or reduce the risk of gastrointestinal disorders and conditions including, but not limited to, constipation disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, post-operative ileus, food allergy or food 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 disease, diarrheal disorders, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis.

[0021] The compounds, compositions, or medicaments can also be used to treat, prevent, ameliorate, control, or reduce the risk of pulmonary diseases and conditions such as chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperreactive airways disorder, and idiopathic pulmonary fibrosis. DETAILED DESCRIPTION OF THE INVENTION

[0022] 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.

[0023] The compound of formula (I) can be used for the treatment, prevention, improvement, control, or reduction of the risk of diseases or disorders involving the EP4 receptor. The compound of formula (I) can be used for the treatment, prevention, improvement, control, or reduction of the risk of gastrointestinal disorders and conditions, including but not limited to constipation disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced bowel disease, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux disease, diarrheal diseases, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis.

[0024] The compounds of formula (I) may also be used to treat, prevent, ameliorate, control, or reduce the risk of pulmonary diseases and conditions such as chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperreactive airways disorder, and idiopathic pulmonary fibrosis.

[0025] Certain novel compounds of the present invention exhibit particularly high activity as EP4 receptor agonists.

[0026] The compounds of the present invention have been demonstrated to have the activity of EP4 receptor agonists.The compounds of the present invention also have low gastrointestinal permeability, as demonstrated by Caco-2 studies.Therefore, it is believed that the compounds of the present invention exhibit low systemic bioavailability when orally administered.The functional agonism of the EP4 receptor expressed in the gastrointestinal tract has the potential to treat various gastrointestinal diseases.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 the cardiovascular side effects resulting from systemic distribution.

[0027] In one aspect, the present invention provides a compound of formula I: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof [In the formula, A is OR', C(O)R', CO2R', C(O)N(R')2, C(O)N(R')S(O)2R 3 , S(O)2R', S(O)2OR', SO2N(R')2, C 1~8 alkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; X is halo, OR', COOR', or C 1~6 is alkyl, L and L' are each independently C 2~4 is alkylene, R 1 H, Halo, CN, NO2, OR', SR', COOR', C 1~6 Alkoxy or C 1~6 is alkyl, R 2 , OR', OC(O)R 3 ,OC(O)OR 3 , CO2R', CON(R')2, SO2N(R')2, SO2R 3 , OSO2R 3 or OSO2N(R')2, R 3is C 1~6 Alkyl, C 3~6 cycloalkyl, or phenyl; R' is H, C 1~6 Alkyl or C 3~6 is cycloalkyl, n is 0, 1, 2, or 3; In each instance, alkyl, alkylene, alkoxy, and cycloalkyl are each optionally and independently selected from OH, SH, CN, NO, COOH, halo, or COOC. 1~4 substituted with up to three of alkyl, In each instance, heterocycloalkyl, aryl, and heteroaryl are each optionally and independently selected from OR′, SR′, CN, NO 2 , CO 2 R′, halo, C 1~4 substituted with up to three of the following: alkyl, or oxo Includes:

[0028] In some embodiments, the compound is a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (Ic), Formula (1), Formula (1a), Formula (1b), Formula (1c), Formula (2a), Formula (2b), Formula (2c), Formula (2d), Formula (3a), Formula (3b), Formula (3c), Formula (3d), Formula (3e), Formula (3f), Formula (5), Formula (5a), Formula (5b), Formula (6), Formula (6a), Formula (6b), or a pharmaceutically acceptable salt or tautomer thereof.

[0029] In some embodiments, the compound is a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (Ic), Formula (1), Formula (1a), Formula (1b), Formula (1c), Formula (2a), Formula (2b), Formula (2c), Formula (2d), Formula (3a), Formula (3b), Formula (3c), Formula (3d), Formula (3e), Formula (3f), Formula (5), Formula (5a), Formula (5b), Formula (6), Formula (6a), Formula (6b), or a pharmaceutically acceptable salt thereof.

[0030] In some embodiments, L' is a group [ka] is.

[0031] In some embodiments, L is a group [ka] is.

[0032] In one embodiment of this aspect, the compound is a compound of formula (1): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

[0033] In some embodiments, A is C(O)OR', C(O)N(R')S(O)R 3 , S(O)2OR', C 1~8 and wherein the heterocycloalkyl and heteroaryl are each optionally and independently selected from OR′, SR′, halo, C 1~4 It is substituted with up to three of the following: alkyl, or oxo.

[0034] In some embodiments, A is chosen from C(O)OR' and heteroaryl.

[0035] In some embodiments, A is selected from the group consisting of: [ka] is selected from.

[0036] In some embodiments, A is [ka] is.

[0037] In some embodiments, A is [ka] is.

[0038] In some embodiments, the compound of Formula (I) is a compound of Formula (Ia), (Ib), or (Ic): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

[0039] In some embodiments, the compound of Formula (I) is a compound of Formula (1a), (1b), or (1c): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

[0040] In some embodiments, the compound of Formula (I) is a compound of Formula (3a), (3b), (3c), (3d), (3e), or (3f): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

[0041] In some embodiments, X is halo or OR'.

[0042] In some embodiments, X is F, Cl, or OH.

[0043] In some embodiments, X is F or OH and n is 0, 1, or 2.

[0044] In some embodiments, X is F, Cl, or OH; and n is 1 or 2.

[0045] In some embodiments, X is F or OH and n is 1 or 2.

[0046] In some embodiments, n is 0.

[0047] In some embodiments, R 1 is H, OH, halo, CN, C optionally substituted with 1 to 3 fluorine atoms 1~6 Alkoxy or C optionally substituted with 1 to 3 fluorine atoms 1~6 It is alkyl.

[0048] In some embodiments, R 1 is H, OH, halo, CN, C optionally substituted with 1 to 3 fluorine atoms 1~6 alkyl or C optionally substituted with 1 to 3 fluorine atoms 1~6 It is alkyl.

[0049] In some embodiments, R 1 is methyl.

[0050] In some embodiments, R 2 is OR', CON(R')2, SO2N(R')2, or OSO2N(R')2.

[0051] In some embodiments, R 2 is OH, CONH2, SO2NH2, or OSO2NH2.

[0052] In some embodiments, R 2 are CON(R')2, SO2N(R')2, OSO2R 3 , or OSO2N(R')2.

[0053] In some embodiments, R 2 is CONH2, SO2NH2, or OSO2NH2.

[0054] In some embodiments, R 2 is CONH2.

[0055] In some embodiments, provided herein are compounds of formula (2a), (2b), (2c), or (2d): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof is provided.

[0056] In some embodiments, R 1 is H or C 1~6 In a further embodiment, R 1 is methyl. In another embodiment, n is 0.

[0057] In some embodiments, provided herein are compounds of formula (5), (5a), (5b), (6), (6a), (6b): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof is provided.

[0058] In some embodiments, R 2 is CO2R', CON(R')2, SO2N(R')2, or OSO2N(R')2. In a further embodiment, R 2 is CONH2, SO2NH2, or OSO2NH2.

[0059] In some embodiments, a compound selected from the group consisting of: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof, or a pharmaceutically acceptable salt thereof, is provided herein.

[0060] In some embodiments, the present invention includes pharmaceutical compositions comprising a compound described herein and a pharmaceutically acceptable excipient.

[0061] 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.

[0062] In some embodiments, 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.

[0063] In some embodiments, the present invention includes a method for treating an EP4 receptor-mediated disease, comprising administering to a patient in need thereof a compound or composition described herein.

[0064] In some embodiments, the EP4 receptor-mediated disease is a gastrointestinal disorder. In a further embodiment, the gastrointestinal disorder is selected from the group consisting of constipation disorder, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorder, 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 disease, diarrheal disease, immune-mediated gastrointestinal disease, 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, hyperreactive airways disorder, and idiopathic pulmonary fibrosis.

[0065] In some embodiments, the compound of Formula I is a compound listed in Table 1 or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

[0066] In some embodiments, a compound selected from the compounds listed in Table 1, or a pharmaceutically acceptable salt thereof: [Table 1-1] [Table 1-2] [Table 1-3] or a pharmaceutically acceptable salt thereof is provided herein. 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 FA formic acid 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 LCMS Liquid Chromatography Mass Spectrometry M mole MeCN acetonitrile MeOH Methanol N standard NBS N-Bromosuccinimide prep HPLC Preparative High Performance Liquid Chromatography RT room temperature sat saturation THF tetrahydrofuran UPLC Ultra High Performance Liquid Chromatography

[0067] definition

[0068] For the purposes of this application, the following definitions apply unless otherwise indicated.

[0069] In connection with the use of the compounds described herein, including compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (Ic), Formula (1), Formula (1a), Formula (1b), Formula (1c), Formula (2a), Formula (2b), Formula (2c), Formula (2d), Formula (3a), Formula (3b), Formula (3c), Formula (3d), Formula (3e), Formula (3f), Formula (5), Formula (5a), Formula (5b), Formula (6), Formula (6a), and Formula (6b), the term "treatment" 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. Thus, the term "treatment" includes both preventative (prophylactic) treatment and treatment when measurable or detectable symptoms of the disease or disorder are present.

[0070] The term "therapeutically effective amount" (e.g., in reference to 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, an effective therapeutic amount is an amount sufficient to provide a desired level of pain relief. The desired level of pain relief can be, for example, a complete elimination of pain or a reduction in the intensity of pain.

[0071] As used herein, the term "hydroxyl" or "hydroxy" refers to an --OH moiety.

[0072] 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, heteroaralkyl carbonylaminoalkylaminocarbonyl, 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.Non-limiting examples of substituted alkyls include carboxyalkyl (such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, aralkyl, (alkoxyaryl)alkyl, (sulfonylamino)alkyl (such as (alkyl-SO-amino)alkyl), aminoalkyl, amidoalkyl, (cycloaliphatic)alkyl, or haloalkyl.

[0073] As used herein, an "alkylene" group refers to a divalent branched or straight-chain alkyl group containing 2 to 12 (e.g., 2 to 8, 2 to 6, or 2 to 4) carbon atoms and serving to link two chemical moieties. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, butylene, isopropylene (methylethylene), and isobutylene (2-methylpropylene). Alkylene groups can be substituted (i.e., optionally substituted) with one or more substituents defined for alkyl groups.

[0074] As used herein, "amide" encompasses both "aminocarbonyl" and "carbonylamino." These terms, when used alone or in combination with other groups, are used in combination with -N(R X )-C(O)-R Y or -C(O)-N(R X )2 and when used internally -C(O)-N(R X )- or -N(R X )-C(O)-(wherein, R X and R Yrefers to an amide group, such as an alkylamide (such as alkylcarbonylamino or alkylaminocarbonyl), (heterocycloaliphatic)amide, (heteroaralkyl)amide, (heteroaryl)amide, (heterocycloalkyl)alkylamide, arylamide, aralkylamide, (cycloalkyl)alkylamide, or cycloalkylamide.

[0075] As used herein, an "amino" group is -NR X R Y In the formula, R X and R Y is independently 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.

[0076] As used herein, the group "aryl," used alone or as part of a larger moiety such as "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 of the rings in the bicyclic or tricyclic ring system is aromatic. Bicyclic and tricyclic groups include benzo-fused 2- to 3-membered carbocyclic rings. For example, benzo-fused groups include groups having two or more C 4~8 Aryl includes phenyl fused to a carbocyclic moiety. Aryl includes 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)carbonyl], cycloaliphatic, hetero ... The aryl may be optionally substituted with one or more substituents including aryl, ((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.

[0077] Non-limiting examples of substituted aryls include haloaryls [e.g., mono-, di(e.g., p,m-dihaloaryls), 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, (( ...

[0078] 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.

[0079] Cycloalkyl groups include 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, 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.

[0080] As used herein, a "heterocycloalkyl" group refers to a 3- to 10-membered mono- or bicyclic (fused or bridged) (e.g., 5- to 10-membered mono- 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 are classified as heteroaryls.

[0081] Heterocycloalkyl groups include 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, or and (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.

[0082] As used herein, a "heteroaryl" group refers to a monocyclic, bicyclic, or tricyclic ring system having 4 to 15 ring atoms, where one or more of the ring atoms is a heteroatom (e.g., N, O, S, or a combination thereof), and the monocyclic ring system is aromatic, or at least one of the rings in the bicyclic or tricyclic ring system is aromatic. Heteroaryl groups include benzo-fused ring systems having two to three rings. For example, benzo-fused groups include benzo (e.g., indolyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furyl, benzo[b]thiophenyl, quinolinyl, or isoquinolinyl) fused with one or two 4- to 8-membered heterocyclic aliphatic moieties. 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.

[0083] Monocyclic heteroaryls include, but are not limited to, furyl, thiophen-yl, 2H-pyrrolyl, pyrrolyl, oxazolyl, triazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4H-pyranyl, pyridyl, pyridazyl, pyrimidyl, pyrazolyl, pyrazyl, or 1,3,5-triazyl. Monocyclic heteroaryls are numbered according to standard chemical nomenclature.

[0084] 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, bexo[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 chemical nomenclature.

[0085] Heteroaryl includes 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, (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, heteroaryl can be unsubstituted.

[0086] 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, ((heterocyclicaliphatic)carbonyl)heteroaryls, and ((alkylcarbonyl)amino)heteroaryls], (cyanoalkyl)heteroaryls, (alkoxy)heteroaryls, (sulfamoyl)heteroaryls, Included are aryl [e.g., (aminosulfonyl)heteroaryl], (sulfonyl)heteroaryl [e.g., (alkylsulfonyl)heteroaryl], (hydroxyalkyl)heteroaryl, (alkoxyalkyl)heteroaryl, (hydroxy)heteroaryl, ((carboxy)alkyl)heteroaryl, (((dialkyl)amino)alkyl]heteroaryl, (heterocyclic aliphatic)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].

[0087] As used herein, an "alkoxy" group refers to an alkyl-O- group, where "alkyl" is defined above.

[0088] As used herein, a "carboxy" group refers to a group such as -COOH, -COOR when used as a terminal group. X , -OC(O)H, -OC(O)R Xand when used as an internal group it refers to -OC(O)- or -C(O)O-.

[0089] As used herein, a "mercapto" group refers to -SH.

[0090] As used herein, a "sulfo" group refers to -SO3H or -SO3R when used terminally. X When used internally it refers to -S(O)3-.

[0091] 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 When used internally, it refers to Structure-NR X -S(O)2-NR Y - where R X , R Y , and R Z is defined above.

[0092] As used herein, a "sulfamoyl" group refers to a group having the structure -OS(O)-NR Y R Z In the formula, R Y and R Z is defined above.

[0093] As used herein, a "sulfonamide" group refers to a group having the structure -S(O)2-NR when used terminally. x R y or -NR x -S(O)2-R z When used internally, it refers to the structure -S(O)2-NR x -or-NR x -S(O)2-, where R x , R y , and R Z is defined above.

[0094] As used herein, a "sulfanyl" group refers to a group such as -SR when used terminally. X When used internally it refers to -S-, where R X is defined above. Examples of sulfanyl include aliphatic-S—, cycloaliphatic-S—, aryl-S—, and the like.

[0095] As used herein, a "halogen" or "halo" group refers to fluorine, chlorine, bromine, or iodine.

[0096] As used herein, "oxo" refers to =O.

[0097] As used herein, the term "vicinal" generally refers to the arrangement of substituents on a group containing two or more carbon atoms, where the substituents are attached to adjacent carbon atoms.

[0098] As used herein, the term "geminal" generally refers to the arrangement of substituents on a group containing two or more carbon atoms, where the substituents are attached to the same carbon atom.

[0099] The terms "terminal" and "internal" 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 the group is 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.

[0100] 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 has the structure -[CH] v-, where 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 the structure -[CQQ] v wherein Q is independently hydrogen or an aliphatic group, provided that at least one Q is an aliphatic group. The term aliphatic chain includes alkyl chains, alkenyl chains, and alkynyl chains, where alkyl, alkenyl, and alkynyl are defined above.

[0101] The phrase "optionally substituted" is used interchangeably herein with the phrase "substituted or unsubstituted." As used herein, the compounds of the present invention can be optionally substituted with one or more substituents as exemplified generally above or by specific classes, subclasses, and species of the present invention. Unless otherwise noted, each specific group of a variable described herein can be optionally substituted with one or more substituents as described herein. Each substituent of a specific group is 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 can be substituted with alkylsulfanyl, and the alkylsulfanyl can 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 form a ring together with the atoms to which they are bound.

[0102] As used herein, the term "substituted," whether preceded by the term "optionally" or not, generally refers to the replacement of a hydrogen atom in a given structure with the radical of a particular substituent. Specific substituents are described in the definitions above and in the description of compounds and examples thereof below. Unless otherwise indicated, an optionally substituted group can have a substituent at each substitutable position of the group, and when more than one position in any given structure can be substituted with more than one substituent selected from a specified group, the substituents can 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 ordinary skill in the art will recognize, the combinations of substituents contemplated by the present invention are those that result in the formation of stable or chemically feasible compounds.

[0103] 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 of the purposes disclosed herein. In some embodiments, a stable or chemically feasible compound is one that does not change substantially when kept at a temperature of 40° C. or less in the absence of moisture or other chemically reactive conditions for at least one week.

[0104] To the extent that any of the compounds described contain chiral centers, the present invention extends to all optical isomers of such compounds, whether in racemic or resolved enantiomeric form. The invention described herein relates to all crystalline forms, solvates, and hydrates of any of the disclosed compounds (however so prepared). To the extent that any of the compounds disclosed herein contain an acid or basic center, such as a carboxylate or amino group, all salt forms of the compound are included herein. The invention described herein further relates to all tautomers of the compounds presented herein. For pharmaceutical use, the salts should be considered to be pharmaceutically acceptable salts.

[0105] The salt or pharmaceutically acceptable salt that can be mentioned includes acid addition salt and base addition salt.Such salt 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, and then removing the solvent or medium using standard techniques (for example, in vacuo, lyophilization, or filtration).Salt can also be prepared by exchanging the counterion of the compound in salt form with another counterion, for example, using a suitable ion exchange resin.

[0106] 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.

[0107] 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, camphoric acid, benzo ... 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., For example, 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 Acid addition salts with carboxylic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, tartaric acid (e.g., (+)-L-tartaric acid), thiocyanic acid, undecylenic acid, and valeric acid are included.

[0108] 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 a solvating solvent) into the solid-state 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 a solvating solvent. Whether a solvate has formed in any given case can be determined by subjecting crystals of the compound to analysis using well-known standard techniques, such as thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and X-ray crystallography.

[0109] Solvates can be stoichiometric or non-stoichiometric solvates. Particular solvates can be hydrates, and examples of hydrates include hemihydrates, monohydrates, and dihydrates. For a more detailed discussion of solvates and the methods used to prepare and characterize them, see Bryn et al., Solid-State Chemistry of Drugs, Second Edition, published by SSCI, Inc. of West Lafayette, IN, USA, 1999, ISBN 0-967-06710-3.

[0110] The term "pharmaceutical composition" in the context of the present invention refers to a composition comprising an active agent and further comprising one or more pharmaceutically acceptable carriers or excipients. Depending on the method of administration and the nature of the dosage form, the composition may contain, 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, lozenges, emulsions, solutions, cachets, granules, capsules, and suppositories, as well as injectable liquid preparations including liposomal preparations.

[0111] The compounds of the present invention may contain one or more isotopic substitutions; a reference to a particular element includes within its scope all isotopes of the element. For example, a reference to hydrogen includes within its scope all isotopes of the element. 1 H, 2 H(D), and 3 Similarly, references to carbon and oxygen include within their scope, respectively, 12 C. 13 C, and 14 C, and 16 O and 18 O. Similarly, reference to a particular functional group also includes isotopic variations within its scope, unless the context indicates otherwise. For example, a reference to an alkyl group such as an ethyl group or an alkoxy group such as a methoxy group also encompasses variations in which one or more of the hydrogen atoms in the group are in the form of a deuterium or tritium isotope, for example, an ethyl group in which all five hydrogen atoms are in the form of a deuterium isotope (a perdeuteroethyl group), or a methoxy group in which all three hydrogen atoms are in the form of a deuterium isotope (a trideuteromethoxy group). Isotopes can be radioactive or non-radioactive.

[0112] Treatment dosage can vary depending on patient requirements, the severity of the condition being treated, and the compound being used.Determining the appropriate dosage for specific circumstances is within the skill of those skilled in the art.Generally, treatment is started with a dosage that is less than the optimal dose of compound.Then, dosage is gradually increased until the optimal effect is achieved under the circumstances.For convenience, if necessary, the total daily dosage can be divided and administered several times throughout the day.

[0113] The magnitude of an effective dose of a compound will, of course, vary depending on the nature and 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 ranges from 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.

[0114] Combination therapy

[0115] An effective amount can be achieved in the methods or pharmaceutical compositions 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 suitable therapeutic agent, e.g., an antiviral agent or a vaccine. When using "combination therapy," an effective amount can be achieved using a first amount of a compound of the invention and a second amount of an additional suitable therapeutic agent.

[0116] 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., an amount that is each therapeutically effective when administered alone). In another embodiment, the compound of the invention and the additional therapeutic agent are each administered in an amount that does not provide a therapeutic effect alone (a subtherapeutic dose). In yet another embodiment, the compound of the invention is 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.

[0117] 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 the terms does not restrict the order in which the therapies (e.g., prophylactic and / or therapeutic agents) are administered to a subject.

[0118] Co-administration includes administering a first amount and a second amount of the co-administered compounds in an essentially simultaneous manner, for example, in a single pharmaceutical composition, for example, in a capsule or tablet having a fixed ratio of the first and second amounts, or in multiple separate capsules or tablets for each. Additionally, such co-administration also includes administering each compound in a sequential manner, in any order.

[0119] In one embodiment, a compound of the present invention and an additional therapeutic agent are administered separately, sequentially, or simultaneously to a subject.

[0120] When co-administration involves separately administering a first amount of the compound of the present invention and a second amount of the additional therapeutic agent, the compounds are administered close enough in time to achieve 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.

[0121] More specifically, a first therapeutic agent (e.g., a prophylactic or therapeutic agent such as a compound of the present invention) can be administered to a subject before (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), simultaneously with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second therapeutic agent (e.g., a prophylactic or therapeutic agent such as an anticancer agent).

[0122] It is understood that the method of co-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, where the combined effect is greater than the additive effect that results from administering the first amount of a compound of the invention and the second amount of an additional therapeutic agent separately.

[0123] As used herein, the term "synergy" refers to a combination of a compound of the present invention and another therapeutic agent (e.g., a prophylactic or therapeutic agent) that is more effective than the additive effects of the therapeutic agents. A synergistic effect of a combination of therapeutic agents (e.g., a prophylactic or therapeutic agent combination) may allow for the use of lower dosages of one or more therapeutic agents or may allow for less frequent administration of the therapeutic agents to a subject. The ability to utilize lower dosages of a therapeutic agent (e.g., a prophylactic or therapeutic agent) and / or administer the therapeutic agent less frequently may reduce the toxicity associated with administering the therapeutic agent to a subject without reducing the efficacy of the therapeutic agent in preventing, managing, or treating a disorder. Furthermore, a synergistic effect may result in improved efficacy of the agents in preventing, managing, or treating a disorder. Finally, a synergistic effect of a combination of therapeutic agents (e.g., a prophylactic or therapeutic agent combination) may avoid or reduce adverse and undesirable side effects associated with using either therapeutic agent alone.

[0124] The presence of synergistic effects can be determined using a suitable method for evaluating drug interactions. Suitable methods include, for example, the sigmoid-Emax equation (Holford, NHG and Scheiner, LB, Clin. Pharmacokinet. 6:429-453 (1981)), the Loewe additivity equation (Loewe, S. and Muischnek, H., Arc h. Exp. Pathol Pharmacol. 114:313-326 (1926)), and the median effect equation (Chou, TC and Talalay, P., Adv. Enzyme Regul. 22:27-55 (1984)). Each of the above-mentioned equations can be applied using experimental data to generate corresponding graphs that are useful for evaluating the effects of drug combinations. The corresponding graphs associated with the above-mentioned equations are concentration-effect curves, isobologram curves, and combination index curves, respectively.

[0125] In one aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, a pharmaceutically acceptable excipient, and at least one additional therapeutic agent. In some embodiments, the compound of the present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and at least one additional therapeutic agent are co-formulated. In some embodiments, the compound of the present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and at least one additional therapeutic agent are formulated separately.

[0126] In another aspect, the present invention provides a kit comprising a compound of the present 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 present invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and at least one additional therapeutic agent to a subject in need thereof.

[0127] 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.

[0128] The compound of the invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer and the additional therapeutic agent may be administered separately, sequentially, or simultaneously to a subject.

[0129] 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 disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, postoperative ileus, food allergy or 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 disease, diarrheal disorders, immune-mediated gastrointestinal disorders, 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, hyperreactive airways disorder, and idiopathic pulmonary fibrosis.

[0130] 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 aminosalicylate 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.

[0131] Pharmaceutical preparations

[0132] While it is possible for the active compound to be administered alone, it may be preferable to administer the active compound as a pharmaceutical composition (eg, a formulation).

[0133] Therefore, in another embodiment of the present invention there is provided a pharmaceutical composition comprising at least one compound of formula (I) as defined above, together with at least one pharmaceutically acceptable excipient.

[0134] When the pharmaceutical composition includes at least one additional therapeutic agent, the compound of the invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and the at least one additional therapeutic agent may be co-formulated, or the compound of the invention, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and the at least one additional therapeutic agent may be formulated separately.

[0135] The composition may be a tablet composition.

[0136] The composition may be a capsule composition.

[0137] Pharmaceutically acceptable excipients can 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), granules, binders, flow aids, coatings, release-controlling agents (e.g., polymers or waxes that delay or retard release), binding agents, disintegrants, buffers, lubricants, preservatives, antifungal and antibacterial agents, antioxidants, buffers, tonicity adjusters, thickeners, flavors, sweeteners, pigments, plasticizers, taste-masking agents, stabilizers, or any other excipient conventionally used in pharmaceutical compositions.

[0138] As used herein, the term "pharmaceutically acceptable" means a compound, material, composition, and / or dosage form that 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 benefit / risk ratio. Each excipient must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

[0139] 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, PA, USA).

[0140] The pharmaceutical compositions may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic, otic, rectal, intravaginal, or transdermal administration.

[0141] Pharmaceutical dosage forms suitable for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), caplets, pills, lozenges, syrups, solutions, powders, granules, elixirs and suspensions, sublingual tablets, wafers, or patches such as buccal patches.

[0142] The tablet composition can contain a unit dose of the 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, e.g., 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 can 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 buffers or citrate buffers), effervescent agents, e.g., citrate / bicarbonate mixtures, etc. Such excipients are well known and need not be discussed in detail here.

[0143] Tablets can be designed to release drug upon contact with gastric fluids (immediate-release tablets) or to release drug in a controlled manner over an extended period of time or in specific areas of the gastrointestinal tract (extended-release tablets).

[0144] Pharmaceutical compositions typically contain about 1% (w / w) to about 95%, preferably % (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% of a pharmaceutically acceptable excipient or a combination of excipients. Pharmaceutical compositions contain about 1% to about 95%, preferably about 20% to 90%, of the active ingredient. Pharmaceutical compositions according to the present invention may be in unit dosage form, for example, in the form of ampoules, vials, suppositories, pre-filled syringes, dragees, powders, tablets, or capsules.

[0145] Tablets and capsules may contain, for example, (depending on the drug dose) 0-20% disintegrant, 0-5% lubricant, 0-5% flow aid, and / or 0-99% (w / w) filler or bulking agent. They may also contain 0-10% (w / w) polymer binder, 0-5% (w / w) antioxidant, and 0-5% (w / w) colorant. Sustained-release tablets may typically further contain (depending on the dose) 0-99% (w / w) release-controlling (e.g., retarding) polymer. The film coat of a tablet or capsule typically contains 0-10% (w / w) polymer, 0-3% (w / w) pigment, and / or 0-2% (w / w) plasticizer.

[0146] Parenteral formulations typically contain (depending on the dose and if lyophilized) 0-20% (w / w) buffer, 0-50% (w / w) cosolvent, and / or 0-99% (w / w) water for injection (WFI). Intramuscular depot formulations may also contain 0-99% (w / w) oil.

[0147] Pharmaceutical formulations may be provided to patients in "patient packs" that contain an entire course of treatment in a single package, usually a blister pack.

[0148] The compounds of formula (I) are generally provided in unit dosage form and, thus, 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 (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.

[0149] For oral compositions, a unit dosage 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.

[0150] The active compound is 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 compound to be administered can be determined by a supervising physician according to standard procedures.

[0151] Synthesis of compounds of formula (1)

[0152] LC / MS method 1

[0153] Instrument: Acquity UPLC equipped with a photodiode array detector and a QDA mass detector; Column: Acquity C-18, 1.6 micron, 50 x 2.1 mm; Gradient [time (min) / solvent B (%) in A]: 0.00 / 10, 0.75 / 10, 2.80 / 90, 4.50 / 100, 4.60 / 100, 4.70 / 10; 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.

[0154] LC / MS method 2

[0155] Instrument: Acquity UPLC equipped with a photodiode array detector and a QDA mass detector; Column: Acquity C-18, 1.6 micron, 50 x 1.6 mm; Gradient [time (min) / solvent B (%) in A]: 0.00 / 3, 0.20 / 3, 2.70 / 98, 3.00 / 100, 3.50 / 100, 3.51 / 3, 4.00 / 3; Solvents: Solvent A = 0.1% formic acid in water; Solvent B = 0.1% formic acid in water / acetonitrile (10:90); Column temperature 35 °C; Flow rate 0.9 mL / min.

[0156] LC / MS method 3

[0157] Instrument: Water 2690 equipped with a photodiode array detector and a QDA mass detector; Column: X-Bridge C-18, 5 micron, 100 x 1.6 mm; Gradient [time (min) / % solvent B in A]: 0.01 / 10, 1.00 / 10, 5.00 / 100, 7.00 / 100, 7.50 / 10, 8.00 / 10; Solvents: Solvent A = 0.1% formic acid and 10 mM ammonium carbonate in water; Solvent B = acetonitrile; Column temperature 35 °C; Flow rate 0.9 mL / min.

[0158] General synthetic strategy

[0159] According to Scheme 1, compounds of formula G-4 can be synthesized by first 1) alkylating tert-butyl carbazate with a bromide compound of formula G-1a, or 2) contacting tert-butyl carbazate with an aldehyde of formula G-1b under reductive amination conditions to give hydrazine compound G-2. Contacting G-2 with the divalent compound 3-bromopropionyl chloride gives the cyclized and boc-protected compound of formula G-3. Deprotection of G-4 gives the compound of formula G-4.

[0160] [ka]

[0161] According to Scheme 2, a compound of formula G-4 is alkylated by conjugate addition with a compound of formula G-5 to give a compound of formula G-6. G-6 can then be coupled with an aryl bromide of formula G-7, where R X and R X’ are each independently a lower alkyl group, or R x and R x’Both G-6 and G-7, together with the boron and oxygen atoms to which they are attached, form a 5- or 6-membered heterocycle. Coupling of G-6 and G-7 is typically carried out in the presence of a palladium catalyst, such as PdCl(dppf)·DCM, to give biaryl compounds of formula G-8. G-8 is further reduced to give compounds of formula G-10. At this point in the synthesis, other functional group interconversions are possible; for example, if A is an ester group, compounds G-8 or G-10 can be further hydrolyzed. Reduction (and / or other functional group interconversions (FGI), e.g., hydrolysis) can also be performed via compounds of formula G-9 prior to Pd-catalyzed coupling, according to Scheme 2.

[0162] [ka]

[0163] According to Scheme 3, phenolic compounds of formula G-11 can be further derivatized to compounds of formula G-12 by reaction with sulfonamidating reagents such as sulfamoyl chloride.

[0164] [ka] [Example]

[0165] 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. In the exemplary chiral separations of racemates in the following examples, "Isomer 1" refers to the isomer that eluted first from the chiral column, and "Isomer 2" refers to the isomer that eluted second from the chiral column.

[0166] Preparation by synthesis of intermediates Intermediate 1: Synthesis of methyl 4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate (Intermediate 1) [ka]

[0167] Step (i): Methyl 4-(2-bromoethyl)benzoate (20.00 g, 82.65 mmol), tert-butyl carbazate (12.01 g, 90.92 mmol), NaHCO (27.78 g, 330.60 mmol), and NaI (1.24 g, 8.27 mmol) were suspended in MeCN (200 mL) at room temperature, and the reaction mixture was stirred at 80 °C for 24 h. The reaction mixture was concentrated in vacuo, and the residue was partitioned between water (1000 mL) and EtOAc (800 mL), and the aqueous layer was further extracted with EtOAc (3 × 300 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by gradient column chromatography (normal phase, silica) eluting with 0% to 18% EtOAc in hexane to give impure product, which was further purified by gradient reverse-phase flash column chromatography (reverse phase, C18 silica) eluting with 0% to 55% MeCN in water to give tert-butyl 2-(4-(methoxycarbonyl)phenethyl)hydrazine-1-carboxylate (5.6 g, 23%) as a yellow oil. LCMS (Method 3) confirmed the product (m / z 239 (ES+, M+H-tBu) at 2.20 min).

[0168] Step (ii): tert-Butyl 2-(4-(methoxycarbonyl)phenethyl)hydrazine-1-carboxylate (5.60 g, 19.04 mmol) was dissolved in MeCN (60 mL), potassium carbonate (13.16 g, 95.19 mmol) was added to the reaction mixture at room temperature, and the reaction mixture was stirred at room temperature for 10 minutes. After this time, 3-bromopropionyl chloride (2.90 mL, 28.56 mmol) was added dropwise at room temperature, and the reaction mixture was stirred at room temperature for 24 hours. The reaction mixture was concentrated in vacuo, and the resulting residue was partitioned between water (800 mL) and EtOAc (500 mL), and the aqueous layer was further extracted with EtOAc (2 x 200 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the product was purified by gradient column chromatography (normal phase, silica) eluting with 0% to 22% EtOAc in hexane to give impure product, which was further purified by gradient reverse-phase flash column chromatography (reverse phase, C18 silica) eluting with 0% to 45% MeCN in water to give tert-butyl 2-(4-(methoxycarbonyl)phenethyl)-3-oxopyrazolidine-1-carboxylate (6.5 g, 98%) as a yellow oil. LCMS (Method 3) (m / z 293 (ES+, M+HtBu) at 2.43 min) confirmed the product.

[0169] Step (iii): tert-Butyl 2-(4-(methoxycarbonyl)phenethyl)-3-oxopyrazolidine-1-carboxylate (6.5 g, 18.7 mmol) was dissolved in dioxane (70 mL), and 4 N HCl in dioxane (70 mL) was added dropwise to the reaction mixture at 0° C. The reaction mixture was stirred at room temperature for 4 hours. The solvent was removed in vacuo to give the crude product, which was purified by trituration with 20% MeOH in diethyl ether to give methyl 4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate (4.8 g, 90%) as an off-white powder. LCMS (Method 3) (m / z 249 (ES+, M+H) at 1.57 min) confirmed the product. 1H NMR:(400MHz,DMSO)δ:2.62-2.58(t,2H,J=8.2Hz),3.06-3.02(t,2H,J=7.4Hz),3.58-3.56(t,2H ,J=4.4Hz),3.74-3.70(t,2H,J=7.2Hz),7.46-7.44(d,2H,J=8.4Hz),7.90-7.89(d,2H,J=8.0Hz).

[0170] Intermediate 2: Synthesis of methyl 4-(2-(2-(3-(3-bromophenyl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoate (Intermediate 2) [ka]

[0171] Step (i): Methyl 4-bromo-2,6-difluorobenzoate (40.00 g, 159.36 mmol), potassium vinyltrifluoroborate (32.00 g, 239.04 mmol), and TEA (42 mL, 318.72 mmol) were dissolved in IPA (400 mL) and purged with nitrogen gas at room temperature for 30 minutes. After this time, PdCl(dppf)·DCM (13.01 g, 15.93 mmol) was added, and the reaction mixture was stirred at 80 °C for 3 hours. The reaction mixture was then partitioned between water (1000 mL) and EtOAc (1000 mL). The aqueous layer was further extracted with EtOAc (2 × 500 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by gradient flash column chromatography (normal phase, silica) eluting with 0% to 5% EtOAc in hexanes to give methyl 2,6-difluoro-4-vinylbenzoate (25.0 g, 79%) as a yellow oil.

[0172] Step (ii) Methyl 2,6-difluoro-4-vinylbenzoate (25.0 g, 126.19 mmol) was dissolved in a mixture of t-butanol (130 mL) and water (175 mL) at room temperature. NBS (26.94 g, 151.43 mmol) was then added portionwise at room temperature and stirred at 40 °C for 16 h. After cooling to 5 °C, NaOH solution (10.09 g, 252.23 mmol) was added and stirred at room temperature for 30 min. The reaction mixture was then partitioned between water (800 mL) and EtOAc (500 mL), and the aqueous layer was further extracted with EtOAc (2 × 300 mL). The organic layers were combined and dried (Na SO ). The solvent was removed in vacuo to give crude methyl 2,6-difluoro-4-(oxiran-2-yl)benzoate (22.8 g, 84%) as a yellow oil.

[0173] Step (iii): Methyl 2,6-difluoro-4-(oxiran-2-yl)benzoate (22.8 g, 106.54 mmol) was dissolved in MeOH (500 mL) and 10% palladium on carbon (6.9 g) containing 50% water was added. After this time, ammonium formate (67.22 g, 1065.42 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 16 hours. Upon completion, the reaction mixture was filtered through a bed of Celite, washed with MeOH (3000 mL), and the filtrate was concentrated in vacuo. The reaction mixture was then partitioned between water (1000 mL) and EtOAc (500 mL). The aqueous layer was further extracted with EtOAc (2 × 300 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo to give the crude product, which was purified by gradient flash column chromatography (normal phase, silica), eluting the product with 0% to 26% EtOAc in hexane to give methyl 2,6-difluoro-4-(2-hydroxyethyl)benzoate (11 g, 47.80%) as a yellow oil.

[0174] Step (iv): Methyl 2,6-difluoro-4-(2-hydroxyethyl)benzoate (11.00 g, 50.87 mmol) was dissolved in DCM (300 mL) at room temperature. After this, Dess-Martin periodinane (32.36 g, 76.31 mmol) was added portionwise at room temperature, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then partitioned between saturated aqueous NaHCO (500 mL) and DCM (800 mL). The aqueous layer was further extracted with EtOAc (2 × 500 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo to give crude methyl 2,6-difluoro-4-(2-oxoethyl)benzoate (7.96 g, 73%) as a white solid. This was used in the next step without purification.

[0175] Step (v): Methyl 2,6-difluoro-4-(2-oxoethyl)benzoate (7.92 g, 37.00 mmol) and tert-butyl carbazate (4.89 g, 37.00 mmol) were dissolved in methanol (100 mL) under a nitrogen atmosphere, and 4 Å molecular sieves were added to maintain moisture-free conditions. Glacial acetic acid (0.3 mL, 3.70 mmol) was then added at room temperature and stirred for 3 hours. The reaction mixture was then cooled to 0 °C, and sodium cyanoborohydride (2.78 g, 44.40 mmol) was added in portions and stirred for 12 hours at room temperature. The reaction mixture was concentrated in vacuo to give a residue. This was partitioned between saturated aqueous NaHCO (500 mL) and EtOAc (300 mL), and the aqueous layer was further extracted with EtOAc (2 × 200 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo to give the crude product, which was purified by gradient column chromatography (normal phase, silica) eluting the product with 0% to 3% EtOAc in DCM to give crude tert-butyl 2-(3,5-difluoro-4-(methoxycarbonyl)phenethyl)hydrazine-1-carboxylate (9.10 g, 65%) as a white solid. LCMS (Method 2) (m / z 275 (ES+, M+H-tBu) at 2.32 min) confirmed the product.

[0176] Step (vi): tert-Butyl 2-(3,5-difluoro-4-(methoxycarbonyl)phenethyl)hydrazine-1-carboxylate (9.00 g, 27.26 mmol) was dissolved in DMF (20 mL), and potassium carbonate (18.83 g, 136.30 mmol) was added to the reaction mixture at room temperature and stirred for 10 minutes. After this, 4-bromobutanoyl chloride (3.4 mL, 40.89 mmol) was added dropwise at room temperature, and the reaction mixture was stirred at room temperature for 3 hours and then at 40° C. for 16 hours. The reaction mixture was partitioned between water (500 mL) and EtOAc (700 mL), and the aqueous layer was further extracted with EtOAc (250 mL × 2). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the product was purified by gradient column chromatography (reverse phase, C18 silica) eluting with 0% to 50% MeCN in water to give (3.5 g, 33%) as a yellow oil. LCMS (Method 2) confirmed the product (m / z 329 (ES+, M+H-tBu) at 2.48 min).

[0177] Step (vii): tert-Butyl 2-(3,5-difluoro-4-(methoxycarbonyl)phenethyl)-3-oxopyrazolidine-1-carboxylate (3.5 g, 9.11 mmol) was dissolved in dioxane (40 mL), and 4 N HCl in dioxane (20 mL) was added dropwise to the reaction mixture at 0° C. The reaction mixture was stirred at room temperature for 18 hours. The solvent was removed in vacuo to give the crude product, which was purified by trituration with 20% MeOH in diethyl ether to give methyl 2,6-difluoro-4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate HCl salt (2.62 g, 79%) as an off-white amorphous powder. LCMS (Method 2) (m / z 285 (ES+, M+H) at 1.67 min) confirmed the product.

[0178] Step (vii): tert-Butyl 2-(3,5-difluoro-4-(methoxycarbonyl)phenethyl)-3-oxopyrazolidine-1-carboxylate (3.5 g, 9.11 mmol) was dissolved in dioxane (40 mL), and 4 N HCl in dioxane (20 mL) was added dropwise to the reaction mixture at 0° C. The reaction mixture was stirred at room temperature for 18 hours. The solvent was removed in vacuo to give the crude product, which was purified by trituration with 20% MeOH in diethyl ether to give methyl 2,6-difluoro-4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate HCl salt) (2.62 g, 79%) as an off-white amorphous powder. LCMS (Method 2) (m / z 285 (ES+, M+H) at 1.67 min) confirmed the product.

[0179] Step (viii): Methyl 2,6-difluoro-4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate (HCl salt) (0.80 g, 2.81 mmol) and 1-(3-bromophenyl)prop-2-en-1-one (1.60 g, 14.08 mmol) were suspended in MeOH (10 mL) at room temperature, and the reaction mixture was stirred at room temperature for 15 minutes. After this time, TEA (0.70 mL, 5.27 mmol) was added at room temperature and stirred at 60° C. for 4 hours. The reaction mixture was partitioned between water (200 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 in vacuo and the crude product was further purified by gradient reverse-phase flash column chromatography (reverse phase, C18 silica) eluting the product with 0% to 58% ACN in water to give methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoate (0.65 g, 47%) as a colorless sticky substance. LCMS (Method 2) (m / z 495 (ES+, M+H) at 2.58 min) confirmed the product.

[0180] Step (ix): Methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoate (0.650 g, 1.31 mmol) was dissolved in ethanol (4 mL) and water (2 mL), CeCl (0.97 g, 3.94 mmol) was added, and the reaction mixture was stirred at 0 °C for 5 min. Sodium borohydride (0.20 g, 0.5.26 mmol) was then added at 0 °C, and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was then partitioned between water (100 mL) and EtOAc (150 mL), and the aqueous layer was further extracted with EtOAc (2 × 50 mL). The organic layers were combined and dried (Na SO ). The solvent was removed in vacuo to give Intermediate 2: methyl 4-(2-(2-(3-(3-bromophenyl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoate (0.620 g, 95%) as a brown solid. LCMS (Method 2) (m / z 497 (ES+, M+H) at 2.04 min) confirmed the product. 1 H NMR:(400Mz,DMSO)δ:1.74-1.73(d,2H,J=4.8Hz),2.88-2.76(m,4H),3.12-3.10(d,2H,J=7.2Hz),3.86(s,3H),4.66(s,1 H),5.44-5.43(d,1H,J=4.4Hz),7.12-7.10(d,2H,J=9.6Hz),7.36-7.27(m,2H),7.43-7.41(d,1H,J=8.0Hz),7.55(s,1H).

[0181] Intermediate B: Synthesis of 1-(3-bromophenyl)prop-2-en-1-one (Intermediate B) [ka] Step (i): 3-Bromobenzaldehyde (12.0 g, 56.6 mmol) was dissolved in diethyl ether (30 mL) at 0 °C, and vinylmagnesium bromide (1 M in THF) (200 mL) was added dropwise at 0 °C under a nitrogen atmosphere and stirred at room temperature for 3 h. The reaction mixture was partitioned between saturated aqueous NH4Cl (1000 mL) and EtOAc (800 mL), and the aqueous layer was further extracted with EtOAc (2 × 300 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo, and the crude product was purified by gradient column chromatography (normal phase, silica). The product was eluted with 0% to 14% EtOAc in hexane to give 1-(3-bromophenyl)prop-2-en-1-ol (A) (12.0 g, 94%) as a colorless oil. Step (ii): 1-(3-Bromophenyl)prop-2-en-1-ol (A) (12.0 g, 57.1 mmol) was dissolved in acetone (30 mL) at room temperature. To this was added Jones reagent (39 mL) dropwise at -25 °C and stirred at -25 °C for 30 minutes. The reaction mixture was then warmed to 0 °C and stirred for 30 minutes. The reaction mixture was partitioned between saturated aqueous NaHCO (100 mL) and EtOAc (800 mL), and the aqueous layer was further extracted with EtOAc (2 × 250 mL). The organic layers were combined and dried (Na SO ). The solvent was removed in vacuo, and the crude product was purified by gradient column chromatography (normal phase, silica). The product was eluted with 0% to 8% EtOAc in hexane to give 1-(3-bromophenyl)prop-2-en-1-one (8.0 g, 97%) as a yellow gum. LCMS (no mass ion, 2.39 min) confirmed the product.

[0182] Intermediate 3: Synthesis of methyl 4-(2-(2-(3-(3-bromophenyl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-hydroxybenzoate (Intermediate 3) [ka]

[0183] Step (i): Methyl 4-bromo-2-methoxybenzoate (30.00 g, 156.07 mmol), potassium vinyltrifluoroborate (41.81 g, 312.16 mmol), and TEA (64.93 mL, 468.21 mmol) were dissolved in isopropyl alcohol (300 mL) and purged with nitrogen gas at room temperature for 30 minutes. After this time, PdCl(dppf)·DCM (20.09 g, 24.60 mmol) was added, and the reaction mixture was stirred at 80 °C for 3 hours. The reaction mixture was then partitioned between water (1200 mL) and EtOAc (700 mL). The aqueous layer was further extracted with EtOAc (2 × 500 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by gradient flash column chromatography (normal phase, silica) eluting with 0% to 14% EtOAc in hexane to give methyl 2-methoxy-4-vinylbenzoate (20.00 g, 84%) as a yellow oil. LCMS (Method 1) confirmed the product (m / z 193 (ES+, M+H) at 2.18 min).

[0184] Step (ii): Methyl 2-methoxy-4-vinylbenzoate (20.00 g, 104.05 mmol) was dissolved in a mixture of t-butanol (15 mL) and water (30 mL) at room temperature. NBS (27.78 g, 156.08 mmol) was then added portionwise at room temperature, and the reaction mixture was stirred at 40 °C for 2 h. The reaction mixture was then partitioned between water (1000 mL) and EtOAc (800 mL). The aqueous layer was further extracted with EtOAc (2 × 300 mL). The organic layers were combined and dried (Na SO ). The solvent was removed in vacuo to give crude methyl 2-methoxy-4-(oxiran-2-yl)benzoate (20.00 g, quantitative yield) as a yellow oil. Note: This product was not stable and was carried to the next step without purification.

[0185] Step (iii): Methyl 2-methoxy-4-(oxiran-2-yl)benzoate (20.00 g, 96.06 mmol) was dissolved in methanol (200 mL) and 10% palladium on carbon (10.00 g) containing 50% water was added. After this, ammonium formate (60.57 g, 960.60 mmol) was added at room temperature, and the reaction mixture was stirred at 60 °C for 4 h. Upon completion, the reaction mixture was filtered through a Celite bed and washed with MeOH (2000 mL). The filtrate was concentrated in vacuo. The crude product was purified by gradient flash column chromatography (normal phase, silica) eluting with 0% to 45% EtOAc in hexane to give crude methyl 4-(2-hydroxyethyl)-2-methoxybenzoate (9.80 g, 49%) as a colorless oil. LCMS (Method 1) confirmed the product (m / z 211 (ES+) at 1.60 min).

[0186] Step (iv): Methyl 4-(2-hydroxyethyl)-2-methoxybenzoate (9.80 g, 46.62 mmol) was dissolved in DCM (100 mL) at room temperature. After this, Dess-Martin periodinane (39.54 g, 93.24 mmol) was added portionwise at room temperature, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was then partitioned between saturated aqueous NaHCO3 (800 mL) and EtOAc (500 mL). The aqueous layer was further extracted with EtOAc (2 x 200 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give methyl 2-methoxy-4-(2-oxoethyl)benzoate (10.0 g, quantitative yield) as a yellow oil, which was used in the next step without further purification.

[0187] Step (v): Methyl 2-methoxy-4-(2-oxoethyl)benzoate (10.0 g, 48.03 mmol) and tert-butyl carbazate (7.62 g, 57.63 mmol) were dissolved in methanol (100 mL) under a nitrogen atmosphere. 4 Å molecular sieves (1.00 g) were also added to maintain moisture-free conditions. Glacial acetic acid (0.3 mL, 4.80 mmol) was then added at room temperature, and the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was then cooled to 0 °C, and sodium cyanoborohydride (4.53 g, 72.05 mmol) was added in several portions. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was filtered under reduced pressure, and the filtrate was concentrated in vacuo. The resulting residue was partitioned between saturated aqueous NaHCO3 (700 mL) and EtOAc (500 mL). The aqueous layer was further extracted with EtOAc (3 × 200 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give the crude product, which was purified by gradient column chromatography (normal phase, silica) eluting the product with 0% to 29% EtOAc in hexane to give crude tert-butyl 2-(3-methoxy-4-(methoxycarbonyl)phenethyl)hydrazine-1-carboxylate (7.0 g, 45%) as a yellow sticky material. LCMS (Method 2) (m / z 347 (ES+, M+Na) at 2.02 min) confirmed the product.

[0188] Step (vi): tert-Butyl 2-(4-(methoxycarbonyl)phenethyl)hydrazine-1-carboxylate (7.00 g, 21.59 mmol) was dissolved in MeCN (70 mL), and potassium carbonate (14.92 g, 107.93 mmol) was added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 10 minutes. After this, 3-bromopropionyl chloride (3.26 mL, 32.39 mmol) was added dropwise at room temperature, and the reaction mixture was stirred at room temperature for 24 hours. After this, the reaction mixture was stirred at 40° C. for 4 hours. The reaction mixture was concentrated in vacuo, and the resulting residue was partitioned between water (700 mL) and EtOAc (500 mL). The aqueous layer was further extracted with EtOAc (2×200 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the product was purified by gradient column chromatography (normal phase, silica) eluting with 0% to 32% EtOAc in hexane to give tert-butyl 2-(3-methoxy-4-(methoxycarbonyl)phenethyl)-3-oxopyrazolidine-1-carboxylate (2.0 g, 24%) as a yellow oil. LCMS (Method 2) (m / z 323 (ES+, M+H-tBu) at 2.27 min) confirmed the product.

[0189] Step (vii): tert-Butyl 2-(3-methoxy-4-(methoxycarbonyl)phenethyl)-3-oxopyrazolidine-1-carboxylate (2.00 g, 5.29 mmol) was dissolved in 1,4-dioxane (20 mL), and 4N HCl in dioxane (20 mL) was added dropwise to the reaction mixture at 0° C. The reaction mixture was stirred at room temperature for 8 hours. The solvent was removed in vacuo to give the crude product, which was purified by trituration with 20% MeOH in diethyl ether to give crude methyl 2-methoxy-4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate HCl salt (1.50 g, 90.36%) as an off-white amorphous powder. LCMS (m / z 279.10 (ES+) at 1.415 min) confirmed the product.

[0190] Step (viii): Methyl 2-methoxy-4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate hydrochloride (1.00 g, 3.18 mmol), 1-(3-bromophenyl)prop-2-en-1-one (3.33 g, 15.88 mmol), and TEA (2.2 mL, 15.88 mmol) were suspended in MeOH (10 mL) at room temperature, and the reaction mixture was stirred at 60 °C for 4 h. The reaction mixture was partitioned between saturated aqueous NH Cl (300 mL) and EtOAc (200 mL). The aqueous layer was further extracted with EtOAc (2 × 100 mL). The organic layers were combined and dried (Na SO ). The solvent was removed in vacuo and the crude product was further purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) eluting the product with 0% to 65% MeCN in water to give methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-methoxybenzoate (1.20 g, 69%) as a colorless sticky substance. LCMS (Method 2) (m / z 489 (ES+, M+H) at 2.38 min) confirmed the product.

[0191] Step (ix): Methyl (R)-4-((1-(3-bromobenzyl)pyrrolidine-2-carboxamido)methyl)-2-methoxybenzoate (0.25 g, 0.51 mmol) was dissolved in DCM (3 mL) and the reaction mixture was cooled to −78° C. After this time, BBr3 (1 M solution in DCM) (2.5 mL, 2.56 mmol) was added and the reaction mixture was stirred at −78° C. for 1 h. The reaction mixture was then partitioned between saturated aqueous NaHCO3 (100 mL) and DCM (70 mL). The aqueous layer was extracted again with EtOAc (2×40 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-hydroxybenzoate (0.18 g, 74%) as a yellow sticky substance. LCMS (Method 2) (m / z 475 (ES+, M+H) at 2.61 min) confirmed the product.

[0192] Step (x): Methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-hydroxybenzoate (0.16 g, 0.34 mmol) was dissolved in ethanol (2 mL) and water (2 mL). To this was added CeCl (0.25 g, 1.01 mmol), and the reaction mixture was stirred at 0 °C for 5 minutes. After this, NaBH (0.085 g, 1.36 mmol) was added at 0 °C, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was partitioned between saturated aqueous NaHCO (30 mL) and EtOAc (30 mL). The aqueous layer was further extracted with EtOAc (2 × 20 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo to give intermediate 3, methyl 4-(2-(2-(3-(3-bromophenyl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-hydroxybenzoate (0.15 g, 93%) as an off-white solid. LCMS (Method 2) (m / z 478 (ES+, M+H) at 2.46 min) confirmed the product.

[0193] Synthetic Preparation of Compounds of Formula I

[0194] Run 1: Synthesis of 4-(2-(2-(3-hydroxy-3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (Compound 1) [ka]

[0195] Step (i): Intermediate 1, methyl 4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate hydrochloride (2.00 g, 7.26 mmol) and 1-(3-bromophenyl)prop-2-en-1-one (1.50 g, 14.52 mmol) were suspended in IPA (20 mL) at room temperature, and the reaction mixture was stirred at room temperature for 15 minutes. After this time, TEA (2.5 mL, 36.30 mmol) was added at room temperature and stirred at 80 °C for 12 hours. The reaction mixture was partitioned between saturated aqueous NH4Cl (500 mL) and EtOAc (300 mL), and the aqueous layer was further extracted with EtOAc (2 × 100 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was further purified by gradient reverse-phase flash column chromatography (reverse phase, C18 silica) eluting the product with 0% to 75% MeCN in water to give methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.80 g, 24%) as a colorless sticky substance. LCMS (Method 3) (m / z 459 (ES+, M+H) at 2.48 min) confirmed the product.

[0196] Step (ii): Methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.25 g, 0.55 mmol), 4-hydroxy-2-methylphenylboronic acid (0.99 g, 0.65 mmol), and potassium carbonate (0.23 g, 1.65 mmol) were suspended in dioxane (3 mL) and water (2 mL) at room temperature, and the reaction mixture was degassed with nitrogen at room temperature for 15 minutes. After this time, PdCl(dppf)·DCM (0.04 g, 0.05 mmol) was added at room temperature, and the reaction mixture was stirred at 100 °C for 4 hours. The reaction mixture was partitioned between water (200 mL) and EtOAc (100 mL), and the aqueous layer was further extracted with EtOAc (2 × 50 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by gradient column chromatography (normal phase, silica) eluting the product with 0% to 80% EtOAc in hexanes to give methyl 4-(2-(2-(3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate Intermediate 4 (0.21 g, 81%) as an off-white solid. LCMS (Method 3) (m / z 509 (ES+, M+Na) at 2.33 min) confirmed the product.

[0197] Step (iii): Methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.21 g, 0.43 mmol) and cerium(iii) chloride (0.32 g, 1.30 mmol) were dissolved in ethanol (3 mL) and water (3 mL) at room temperature, and the reaction mixture was stirred at room temperature for 5 minutes. After this time, sodium borohydride (0.65 g, 1.73 mmol) was added at 0° C., and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was partitioned between saturated aqueous NaHCO (100 mL) and EtOAc (50 mL), and the aqueous layer was further extracted with EtOAc (2×30 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo to give methyl 4-(2-(2-(3-hydroxy-3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.16 g, 76%) as an off-white solid. LCMS (Method 3) (m / z 489 (ES+, M+H) at 2.19 min) confirmed the product.

[0198] Step (iv): Methyl 4-(2-(2-(3-hydroxy-3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.16 g, 0.33 mmol) was dissolved in dioxane (2 mL) and water (2 mL). To this was added LiOH monohydrate (0.073 g, 1.75 mmol) at room temperature and stirred at room temperature for 3 hours. The reaction mixture was then partitioned between water (80 mL) and EtOAc (2 x 50 mL). The aqueous layer was further acidified with 1 N aqueous HCl (approximately 20 mL) to adjust the pH up to approximately 3 and extracted with EtOAc (2 x 70 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give 4-(2-(2-(3-hydroxy-3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid) (0.125 g, 81%) as a brown solid. LCMS (Method 3) (m / z 475 (ES+, M+H) at 1.97 min) confirmed the product. 1 H NMR:(400MHz,DMSO)δ:1.26-1.23(m,3H),1.82-1.80(d,2H,J=6.4Hz),2.16 (s,3H),2.88-2.85(t,3H,J=6.0Hz),3.17(s,3H),4.08(s,1H),4.71-4.69( d,1H,J=6.0Hz),6.87-6.83(m,2H),7.01-6.99(d,1H,J=8.4Hz),7.16-7.14 (d,1H,J=7.6Hz),7.37-7.26(m,5H),7.85-7.83(d,2H,8.4Hz),9.43(s,2H).

[0199] Compounds 1R and 1S: Racemic compound 1 (0.12 g) was enantiomerically separated on a Shimadzu preparative HPLC system (PHENOMNENEX AMYLOSE-250 × 21.2 mm, 5 μm column) using a mobile phase of 10 mM DEA (43.0%) and IPA (57.0%) in heptane (no gradient, flow rate 16.00 mL / min). Isomer 1: (0.004 g, 2.6%, off-white solid). LCMS (Method 3) (m / z 475 (ES+, M+H) at 2.30 min) confirmed the product. 1 H NMR:(400MHz,MeOD)δ:2.03-1.93(m,2H),2.93-2.90(t,4H,J=6.8Hz),3.07-3.03(t, 3H,J=7.2Hz),3.25(s,2H),3.37-3.32(m,1H),3.59-3.56(m,1H),3.71-3.69(m,2H), 4.87-4.84 (t, 1H, J = 6.0 Hz), 6.68-6.66 (m, 1H), 6.73-6.72 (d, 1H, J = 2.0z), 7.02-7.00 (d, 1H, J = 8.0 Hz), 7.25-7.19 (m, 3H), 7.41-7.32 (m, 5H), 7.91-7.89 (d, 2H, J = 8.0 Hz). Isomer 2: Product confirmed by LCMS (Method 3) (m / z 475 (ES+, M+H) at 2.30 min). 1 H NMR:(400MHz,MeOD)δ:1.97-1.94(t,2H,J=6.4Hz),2.93-2.90(t,4H,J=6.8Hz),3.07-3.03(m,3H),3.26(s,2H),3.59-3.57(t,2H J=4.6Hz),3.71-3.69(t,2H J=4.6Hz),4.87-4.84(t,1H,J=6.4Hz),6.68-6.65(dd,1H J=2.4Hz and J=4.0Hz), 6.72-6.72(d,1H,J=2.0Hz),7.02-7.00(d,1H,J=8.0Hz),7.24-7.19(m,3H),7.41-7.32(m,5H),7.91-7.89(d,2H,J=8.0Hz).

[0200] Example 2: Synthesis of 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (Compound 2) [ka]

[0201] Step (i): Intermediate 1, methyl 4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzoate hydrochloride (2.00 g, 7.26 mmol) and 1-(3-bromophenyl)prop-2-en-1-one (1.50 g, 14.52 mmol) were suspended in IPA (20 mL) at room temperature, and the reaction mixture was stirred at room temperature for 15 minutes. After this time, TEA (2.5 mL, 36.30 mmol) was added at room temperature and stirred at 80 °C for 12 hours. The reaction mixture was partitioned between saturated aqueous NH4Cl (500 mL) and EtOAc (300 mL), and the aqueous layer was further extracted with EtOAc (2 × 100 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was further purified by gradient reverse-phase flash column chromatography (reverse phase, C18 silica) eluting the product with 0% to 75% MeCN in water to give methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.80 g, 24%) as a colorless sticky substance. LCMS (Method 3) (m / z 459 (ES+, M+H) at 2.48 min) confirmed the product.

[0202] Step (ii): Methyl 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.60 g, 1.30 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (1.36 g, 5.23 mmol), and K2CO3 (0.54 g, 3.92 mmol) were dissolved in a mixture of dioxane:water (3:2, 5 mL) and purged with nitrogen gas at room temperature for 20 minutes. After this time, PdCl2(dppf)·DCM (0.21 g, 0.26 mmol) was added, and the reaction mixture was stirred at 80 °C for 8 hours. The reaction mixture was then partitioned between water (100 mL) and EtOAc (100 mL), and the aqueous layer was further extracted with EtOAc (2 × 70 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by gradient flash column chromatography (reverse phase, C18 silica) eluting with 0% to 38% MeCN in water to give methyl 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.60 g, 90%) as a white solid. LCMS: (Method 3): m / z 536 (ES+, M+Na) at 2.16 min confirmed the product.

[0203] Step (iii): Methyl 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.60 g, 1.16 mmol) was dissolved in ethanol (3 mL) and water (2 mL), and CeCl (0.86 g, 3.50 mmol) was added to the reaction mixture. The reaction mixture was stirred at 0 °C for 5 min. After this time, sodium borohydride (0.17 g, 4.67 mmol) was added at 0 °C, and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was partitioned between saturated aqueous NaHCO (100 mL) and EtOAc (80 mL), and the aqueous layer was further extracted with EtOAc (2 × 50 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by gradient flash column chromatography (reverse phase, C18 silica), eluting the product with 0% to 35% MeCN in water to give pure methyl 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.50 g, 83%) as an off-white solid. LCMS: (LCMS Method 3): m / z 516 (ES+, M+H) at 2.78 min confirmed the product.

[0204] Step (iv): Methyl 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (1.80 g, 3.49 mmol) was dissolved in dioxane (15 mL) and water (8 mL). LiOH monohydrate (0.73 g, 17.46 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was then partitioned between water (250 mL) and EtOAc (2 x 350 mL). The aqueous layer was further acidified with 4 N aqueous HCl (60 mL) to adjust the pH up to approximately 1 and extracted with EtOAc (2 x 300 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give pure 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (1.47 g, 83%) as an off-white solid. LCMS (Method 2): m / z 502 (ES+ at 1.53 min, M+H) confirmed the product. 1 H NMR:(400MHz,DMSO)δ:1.76-1.84(m,2H,),2.26(s,3H),2.70-2.90(m,4H),3.18(s,3H),4.72(brs,1H),5.37(d, 1H, J=4.0Hz), 7.21-7.45 (m, 8H), 7.74 (dd, 1H, J=7.9 and 1.9Hz), 7.80-7.86 (m, 3H), 7.98 (s, 1H), 12.86 (brs, 1H). Compounds 2R and 2S: Racemic compound 2 was enantiomerically separated on a Waters preparative HPLC system using the following method: column ADH-9.5 x 250 mm 5 micron, solvent A: 0.1% TFA in n-heptane, B: IPA:methanol (70:30) isocratic 75% A / 25% B, variable flow rate (time / flow rate, 0.01 / 4.0, 5.00 / 4.0, 10.00 / 8.0, 100.00 / 8.0, min / ml per min). Isomer 1 (0.489 g, 28%) as an off-white solid. LCMS: (Method 2): m / z 502 (ES+, M+H) at 1.56 min confirmed the product. 1H NMR: (400 MHz, DMSO): δ 1.77-1.86 (m, 2H), 2.27 (s, 3H), 2.74-2.90 (m, 4H), 3.14 (brs, 2H), 4.73 (t, 1H, J = 6.4 Hz), 7.21-7.47 (m, 8H), 7.75 (dd, 1H, J = 7.9 Hz, 1.9 Hz), 7.80-7.86 (m, 3H), 7.98 (s, 1H), 12.86 (brs, 1H). (Note: Some aliphatic protons may be hidden by DMSO and / or water.) Isomer 2 (0.477 g, 27%) as an off-white solid. LCMS: (Method 2): m / z 502 (ES+, M+H) at 1.56 min confirmed the product. 1H NMR: (400 MHz, DMSO) δ 1.77-1.86 (m, 2H), 2.27 (s, 3H), 2.76-2.90 (m, 4H), 3.14 (brs, 2H), 4.69-4.76 (m, 1H), 5.37 (brs, 1H), 7.22-7.45 (m, 8H), 7.75 (dd, 1H, J = 7.9 Hz, 1.9 Hz), 7.81-7.86 (m, 3H), 7.97 (s, 1H), 12.84 (brs, 1H). (Note: Some aliphatic protons may be hidden by DMSO and / or water.)

[0205] Example 3: Synthesis of 4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (Compound 3)

[0206] 3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (3-methyl-4-bromobenzenesulfonamide) was synthesized from 3-methyl-4-bromobenzsulfonamide, PdCl(dppf)·DCM, and KOAc in dioxane (80 °C, 2 h) using the procedure described in Example 1. Reverse-phase flash chromatography in the final step afforded 4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.027 g, 28%) as an off-white solid. LCMS (Method 3) confirmed the product (m / z 538 (ES+, M+H) at 1.90 min). 1 H NMR:(400MHz,DMSO)δ:1.10-1.07(t,1H,J=7Hz),1.82-1.80(d,2H,5.6Hz),2.08(s,1H),2.29(s,4H),2.87-2.85(d,4H,6.4Hz),3.13(s,3H),4.73 (s,1H)5.39-5.38(d,1H,4.4Hz),7.35-7.24(m,3H),7.46-7.37(m,6H),7 .71-7.68(m,1H),7.75(s,1H),7.84-7.82(d,2H,J=8.4Hz),12.85(s,1H).

[0207] Compounds 3R and 3S: Racemic compound 3 was separated enantiomerically on a Waters preparative HPLC system (CROMEGACHIRAL CCO 2 50 × 20 mm, 5 μm column) using a mobile phase of 0.1% FA (75%) in heptane and a mixture (25%) of IPA and methanol (50:50) at a flow rate of 18.00 mL / min without a gradient. Isomer 1: (0.019 g, 6.5%, off-white solid). LCMS (Method 3) confirmed the product (m / z 538 (ES+, M+H) at 1.90 min). 1H NMR:(400MHz,DMSO)δ:1.23(s,2H),1.82-1.81(d,2H,J=6.4Hz),2.33-2.29(m,4H),2.83(s,4H),3.16(s,5H),4.74(s,1H),5 .40(s,1H),7.25-7.23(m,3H),7.44-7.34(m,6H),7.71-7.68(m,1H),7.75-7.75(d,1H,J=1.6Hz),7.82-7.8-(d,2H,J=8Hz). Isomer 2: (0.017g, 5.8%, off-white solid). The product was confirmed by LCMS (Method 3) (m / z 538 (ES+, M+H) at 1.93 min). 1 H NMR:(400MHz,DMSO)δ:1.23-1.16(d,3H,J=28.0Hz),1.82(s,2H),2.29(s,4H),2.82(s,4H),3.14( s,2H),4.74(s,1H),5.39(s,1H)7.24-7.21(d,3H,J=12Hz),7.42-7.35(q,5H),7.80-7.69(m,4H).

[0208] Example 4: Synthesis of 4-(2-(2-(3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (Compound 4) [ka]

[0209] Step (i): Intermediate 4 of Example 1 (methyl 4-(2-(2-(3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate, 0.17 g, 0.34 mmol) was dissolved in dioxane (3 mL) and water (2 mL). LiOH monohydrate (0.073 g, 1.74 mmol) was added at room temperature and stirred at room temperature for 3 hours. The reaction mixture was then partitioned between water (80 mL) and EtOAc (2 x 50 mL). The aqueous layer was further acidified with 4 N aqueous HCl (3 mL) to adjust the pH up to approximately 1 and extracted with EtOAc (2 x 50 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give pure 4-(2-(2-(3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.13 g, 79%) as a white solid. LCMS (Method 1) (m / z 473 (ES+, M+H) at 2.13 min) confirmed the product.

[0210] Step (ii): 4-(2-(2-(3-(4'-hydroxy-2'-methyl-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.130 g, 0.27 mmol) was dissolved in DMA (3 mL). Sulfamoyl chloride (0.095 g, 0.82 mmol) was added at room temperature and stirred at room temperature for 3 hours. The reaction mixture was then partitioned between water (70 mL) and EtOAc (30 mL). The aqueous layer was further extracted with EtOAc (2 x 20 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by gradient flash column chromatography, eluting the product with 0% to 5% methanol in DCM to give pure 4-(2-(2-(3-(2'-methyl-4'-(sulfamoyloxy)-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.15 g, 99%) as a white solid. LCMS (Method 1) (m / z 552 (ES+) at 2.14 min) confirmed the product.

[0211] Step (iiia) (racemic method): 4-(2-(2-(3-(2'-methyl-4'-(sulfamoyloxy)-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.03 g, 0.054 mmol) was dissolved in methanol (3 mL) and 10% palladium on carbon (50% wet) (0.02 g) was added. H2 gas was purged through the reaction mixture at room temperature for 2 hours. Upon completion, the reaction mixture was filtered through a bed of Celite, washed with MeOH (30 mL), and the filtrate was concentrated in vacuo. The crude product was purified by gradient flash column chromatography (reverse phase, C18 silica) eluting the product with 0% to 35% MeCN in water to give 4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-(sulfamoyloxy)-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.010 g, 33%) as a brown solid. LCMS (Method 1) (m / z 554 (ES+, M+H) at 2.01 min) confirmed the product. 1 H NMR:(400Mz,MeOD):1.98-1.95(t,3H,J=6.4Hz),2.26(s,3H),2.95-2.92(m,3H,J=6.8Hz),3.26(s,2H),3.36(s,1H),3.58-3.56(m, 1H),3.70-3.68(m,1H),4.87-4.85(d,2H,J=6.4Hz),7.25-7.12(m,5H),7.25(s,1H),7.46-7.39(m,2H),7.92(s,2H),12.72(s,1H).

[0212] Step (iiib) (chiral separation): 4-(2-(2-(3-(2'-methyl-4'-(sulfamoyloxy)-[1,1'-biphenyl]-3-yl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.10 g, 0.18 mmol) was dissolved in ethanol (3 mL) and water (2 mL), and CeCl (0.13 g, 0.54 mmol) was added to the reaction mixture. The reaction mixture was stirred at 0 °C for 5 min. After this time, sodium borohydride (0.027 g, 0.72 mmol) was added at 0 °C, and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was then partitioned between water (30 mL) and EtOAc (2 × 20 mL). The aqueous layer was further acidified with 4 N aqueous HCl (3 mL) to adjust the pH up to approximately 1 and extracted with EtOAc (2 × 50 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give the crude product.

[0213] Compounds 4R and 4S: Racemic compound 4 was separated enantiomerically on a Schimadzu preparative HPLC system (CROMEGACHIRAL CCJ 25 cm × 20 mm, 5 μm column) using a mobile phase of 0.1% FA (60%) in heptane and a mixture (40%) of IPA and methanol (60:40) at a flow rate of 19.00 mL / min without gradient. Isomer 1: (0.0057 g, 24%, white solid). LCMS (Method 1) confirmed the product (m / z 554 (ES+, M+H) at 2.02 min). Chiral HPLC confirmed the purity of the product at 16.23 min. 1H NMR:(400MHz,MeOD)δ:1.98-1.93(m,2H),2.27(s,3H),2.97-2.91(m,4H),3.2 7-3.27(m,2H),4.88-4.85(t,2H,J=6.4Hz),7.23-7.18(m,4H),7.34-7.31(m, 3H),7.46-7.41(m,2H),7.93-7.92(d,2H,J=6.8Hz)7.63-7.61(d,2H,J=8.8Hz ),7.73-7.71(d,1H,J=7.6Hz),7.83-7.78(m,3H),8.58-8.56(t,1H,J=5.2Hz). Isomer 2: (0.0047 g, 26%, white solid). Product confirmed by LCMS (Method 1) (m / z 554 (ES+, M+H) at 2.02 min). Chiral HPLC: Product purity confirmed at 10.62 min. 1 H NMR:(400MHz,MeOD)δ:1.98-193(m,3H),2.27(s,3H),2.97-2.93(m,4H),3.27(s,2H),4.88-4.85(t,2H,J=6.4Hz ),7.24-7.25(m,4H),7.31-7.29(m,2H),7.34(s,1H),7.46-7.39(m,2H)7.93-7.91(d,2H,J=7.6Hz),8.51(s,1H).

[0214] Example 5: Synthesis of 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoic acid (Compound 5) [ka]

[0215] Step (i): Intermediate 2, methyl 4-(2-(2-(3-(3-bromophenyl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoate (0.20 g, 0.60 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (0.21 g, 0.79 mmol), and KCO (0.17 g, 0.130 mmol) were dissolved in a mixture of dioxane:water (1:1, 5 mL) and purged with nitrogen gas at room temperature for 30 minutes. After this time, PdCl(dppf)·DCM (0.099 g, 0.12 mmol) was added, and the reaction mixture was stirred at 80 °C for 1 hour. The reaction mixture was then partitioned between water (70 mL) and EtOAc (70 mL), and the aqueous layer was further extracted with EtOAc (2 × 30 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo, and the crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica), eluting with 0% to 28% MeCN in water to give methyl 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoate (0.29 g, 87%) as a light brown solid. LCMS (Method 2) (m / z 552 at 1.92 min (ES+, M+H)) confirmed the product.

[0216] Step (ii): Methyl 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoate (0.29 g, 0.29 mmol) was dissolved in dioxane (4 mL) and water (2 mL). LiOH (0.11 g, 0.88 mmol) was added at room temperature and stirred at room temperature for 16 hours. The reaction mixture was acidified with 4 N aqueous HCl (4 mL) to adjust the pH to approximately 1 and extracted with EtOAc (40 mL). The aqueous layer was further extracted with EtOAc (3 x 30 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by reverse-phase gradient flash column chromatography (reverse phase, C18 silica) eluting the product with 0% to 23% ACN in water to give 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2,6-difluorobenzoic acid (0.15 g, 53%) as an off-white solid. LCMS (Method 2) (m / z 538 (ES+, M+H) at 1.49 min) confirmed the product. 1 H NMR:(400MHz,DMSO)δ:1.23(s,2H),1.81-1.80(d,2H,J=4.8Hz),2.26(s,3H),2 .75(s,3H),3.17-3.16(d,4H,3.6Hz),4.12-4.11(d,1H,J=4.8Hz),4.71(s,1H), 5.47-5.46(d,1H,J=3.6Hz),6.75-6.73(d,2H,J=7.2Hz),7.25-7.22(m,2H),7.3 7-7.33(m,2H),7.43-7.39(m,2H),7.76-7.74(m,1H),7.82(s,1H),8.02(s,1H).

[0217] Compounds 5R and 5S: The racemic mixture (70 mg) was enantiomerically separated on a Shimadzu preparative HPLC system (CHROMEGACHIRAL CCO 250 × 25 mm, 5 μm column) using a mobile phase of 0.1% FA (80%) in heptane and a mixture (20%) of IPA and methanol (70:30) without gradient and at a flow rate of 20.00 mL / min. Isomer 1: (0.038 g, 13%, white solid). The product was confirmed by LCMS (Method 2) (m / z 538 (ES+, M+H) at 1.59 min). Chiral HPLC: The purity of the product was confirmed at 14.48 min. 1 H NMR:(400MHz,DMSO)δ:1.81-1.80(d,2H,J=6.4Hz),2.27(s,4H),2.85(s,4H),3.13-3.12(d,2H,J=5.6Hz),4.73(s,1H),5.36 (s,1H),7.10-7.04(m,2H),7.27-7.22(m,2H),7.43-7.34(m,4H),7.76-7.74(m,1H),7.82(s,1H),7.97(s,1H),13.78(s,1H). Isomer 2: (0.052g, 18%, off-white solid). The product was confirmed by LCMS (Method 2) (m / z 538 (ES+, M+H) at 1.60 min). Chiral HPLC: The purity of the product was confirmed at 10.50 min. 1 H NMR:(400MHz,DMSO)δ:1.81-1.80(d,2H,J=5.6Hz),2.27(s,3H),2.85(s,3H),3.14-3.12(d,2H,7.6Hz),4.72(s,1H),5.37(s,1H) ),7.06-7.04(d,2H,J=8.8Hz),7.27-7.22(m,2H),7.43-7.34(m,4H),7.76-7.74(m,1H),7.82(s,1H),7.97(s,1H),13.79(s,1H).

[0218] Example 6: Synthesis of 2-hydroxy-4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (Compound 6) [ka]

[0219] Step (i): Intermediate 3, methyl 4-(2-(2-(3-(3-bromophenyl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-hydroxybenzoate (0.15 g, 0.32 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (0.14 g, 0.47 mmol), and K2CO3 (0.89 g, 0.64 mmol) were dissolved in 4 mL of a 1:1 dioxane:water mixture. Nitrogen gas was purged at room temperature for 30 minutes. After this time, PdCl2(dppf)·DCM (0.026 g, 0.032 mmol) was added, and the reaction mixture was stirred at 80 °C for 1 hour. The reaction mixture was then partitioned between water (50 mL) and EtOAc (50 mL). The aqueous layer was further extracted with EtOAc (2 x 20 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo, and the crude product was purified by gradient flash column chromatography (reverse phase, C18 silica) eluting with 0% to 55% MeCN in water to give methyl 2-hydroxy-4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.13 g, 73%) as a white solid. LCMS (Method 2) (m / z 568 (ES+) at 1.86 min) confirmed the product.

[0220] Step (ii): Methyl 2-hydroxy-4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.13 g, 0.23 mmol) was dissolved in 4 mL of a 1:1 dioxane:water mixture at room temperature, LiOH monohydrate (0.048 g, 1.15 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was partitioned between water (30 mL) and EtOAc (2 x 50 mL). The aqueous layer was further acidified with 4 N aqueous HCl (2 mL) to adjust the pH to approximately 1 and then extracted with EtOAc (2 x 50 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give pure 2-hydroxy-4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.050 g, 39%) as an off-white solid. LCMS (Method 2) (m / z 554 (ES+, M+H) at 1.90 min) confirmed the product.

[0221] Compounds 6R and 6S: Racemic compound 6 (50 mg) was enantiomerically separated on a Waters preparative HPLC system (CHIRALPAK AD-H250 × 10 mm 5 μm column) using a mobile phase of 0.1% FA (80%) in heptane and IPA:acetonitrile (70:30) (20%) without a gradient and at a flow rate of 7.00 mL / min. Isomer 1: (0.006 g, 4.7%, sticky solid). LCMS (Method 2) confirmed the product (m / z 554 (ES+, M+H) at 1.86 min). Chiral HPLC: The purity of the product was confirmed at 18.45 min. 1H NMR:(400MHz,MeOD)δ:1.971-1.955(d,3H,J=6.4Hz),2.331(s,3H),2.908-2.853(m,4H),3.59-3.56(t,3H,J=4.8Hz),3.70-3.68(t,3H,J= 4.6Hz),6.81(s,2H),7.27-7.25(d,1H,J=7.2Hz),7.38-7.36(d,2H,J=8.0Hz),7.49-7.43(m,2H),7.78-7.76(d,2H,J=7.6Hz),7.84(s,1H). Isomer 2: (0.007g, 4.0%, sticky solid). The product was confirmed by LCMS (Method 3) (m / z 554 (ES+, M+H) at 1.87 min). Chiral HPLC: The purity of the product was confirmed at 29.47 min. 1 H NMR:(400MHz,MeOD)δ:1.99-1.96(m,3H),2.33-2.30(d,3H,J=10.4Hz),2.91-2.86(m,4H),3.58-3.56(t,2H,J=4.6Hz),3.70-3.68(t,2H,J= 4.6Hz),6.82(s,2H),7.27-7.25(d,1H,J=7.2Hz),7.38-7.36(d,2H,J=8.0Hz),7.49-7.43(m,2H),7.78-7.76(d,2H,J=8.0Hz),7.83(s,1H).

[0222] Example 7: Synthesis of 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-hydroxybenzoic acid (Compound 7)

[0223] Compound 7 was synthesized by the method described in Example 6 using 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide to give 4-(2-(2-(3-(4'-carbamoyl-2'-methyl-[1,1'-biphenyl]-3-yl)-3-hydroxypropyl)-5-oxopyrazolidin-1-yl)ethyl)-2-hydroxybenzoic acid (0.075 g, 86%) as an off-white solid. LCMS (Method 2) (m / z 518 (ES+, M+H) at 1.62 min) confirmed the product.

[0224] Compounds 7R and 7S: Racemic compound 7 (75 mg) was separated on a Waters preparative HPLC system (CHIRALPAK AD-H250 × 10 mm 5 μm column) using a mobile phase of 0.1% FA (75%) in heptane and a mixture (25%) of IPA and acetonitrile (70:30) at a flow rate of 8.00 mL / min without a gradient. Isomer 1: (0.016 g, 19%, off-white solid). LCMS (Method 2) confirmed the product (m / z 518 (ES+, M+H) at 1.74 min). Chiral HPLC confirmed the purity of the product at 11.21 min. 1 H NMR:(400MHz,DMSO)δ:1.81-1.80(d,2H,J=6.0Hz),2.27(s,3H),2.54(s,1H),2.80-2.7 (d,4H,J=6.4Hz),3.153(s,3H),4.74-4.71(t,1H,J=6.0Hz),5.32(s,1H),6.78-6.74(m , 2H) 7.27-7.22 (m, 2H), 7.36-7.33 (m, 2H), 7.43-7.38 (m, 2H), 7.68-7.66 (d, 1H, J = 8.0 Hz), 7.76-7.73 (dd, 1H, J = 1.6 Hz), 7.81 (s, 1H), 7.97 (s, 1H), 11.20 (s, 1H), 13.84 (s, 1H). Isomer 2: (0.025 g, 29%, off-white solid). Product was confirmed by LCMS (Method 2) (m / z 518 (ES+, M+H) at 1.73 min). Chiral HPLC: Product purity was confirmed at 9.72 min. 1H NMR:(400MHz,DMSO)δ:1.81-1.80(d,2H,J=6.4Hz),2.27(s,4H),2.80(s,4H),3.15-3.14( d,3H,J=6.4Hz),4.74-4.72(d,1H,J=6.0Hz),5.31(s,1H),6.76-6.74(d,1H,J=8.4Hz),6.7 8(s,1H),7.27-7.22(m,2H),7.36-7.33(m,2H),7.43-7.38(m,2H),7.68-7.66(d,1H,J=8. 0Hz), 7.76-7.73(dd,1H,J=1.6Hz),7.81(s,1H),7.97(s,1H),11.20(s,1H),13.84(s,1H).

[0225] Example 8: Synthesis of 2,6-difluoro-4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (Compound 8) Compound (8) was synthesized using 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide by the method described in Example 5 to give 2,6-difluoro-4-(2-(2-(3-hydroxy-3-(2'-methyl-4'-sulfamoyl-[1,1'-biphenyl]-3-yl)propyl)-5-oxopyrazolidin-1-yl)ethyl)benzoic acid (0.040 g, 23%) as a white solid. LCMS (Method 2) (m / z 574 (ES+, M+H) at 1.58 min) confirmed the product.

[0226] Compounds 8R and 8S: Racemic compound 8 (40 mg) was enantiomerically separated on a Shimadzu preparative HPLC system (CHROMEGACHIRAL CCO 250 × 25 mm, 5 μm column) using a mobile phase of 0.1% FA (80%) in heptane and a mixture (20%) of IPA and acetonitrile (70:30) without a gradient and at a flow rate of 22.00 mL / min. Isomer 1: (0.010 g, 5.7%, sticky gum). LCMS (Method 2) confirmed the product (m / z 574 (ES+, M+H) at 1.58 min). Chiral HPLC confirmed the purity of the product at 28.03 min. 1 H NMR: (400 MHz, MeOD) δ: 1.30 (s, 3H), 1.98-1.96 (d, 3H, J = 6.0 Hz), 2.34 (s, 4H), 2.96-2.93 (t, 6H, J = 6.6 Hz), 6.98-6.95 (d, 2H, J = 9.2 Hz), 7.28-7.25 (m, 1H), 7.38-7.36 (m, 2H), 7.50-7.43 (m, 2H), 7.78-7.76 (dd, 1H, J = 1.5 Hz), 7.84-7.84 (m, 1H). Isomer 2: (0.012 g, 6.8%, sticky gum). LCMS (Method 2) confirmed the product (m / z 574 at 1.58 min (ES+, M+H)). Chiral HPLC: The purity of the product was confirmed at 20.05 min. 1 H NMR:(400MHz,MeOD)δ:1.30(s,3H),1.97-1.96(d,2H,J=5.6Hz),2.34-2.32(m,3H),2.96-2.93(t,4H,J=6.4Hz),6.98-6. 95(d,2H,J=9.2Hz),7.28-7.26(m,1H),7.38-7.36(m,2H),7.48-7.45(m,2H),7.78-7.76(dd,1H,J=1.5Hz),7.84(s,1H).

[0227] Example 9: Synthesis of (3'-(3-(2-(4-(1H-tetrazol-5-yl)phenethyl)-3-oxopyrazolidin-1-yl)-1-hydroxypropyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 9) [ka]

[0228] Step (i): 1-Bromo-4-(2-bromoethyl)benzene (10.00 g, 38.18 mmol), tert-butylhydrazine carboxylate (7.57 g, 57.27 mmol), NaHCO (12.84 g, 15.27 mmol), and NaI (0.57 g, 3.82 mmol) were suspended in MeCN (100 mL) at room temperature, and the reaction mixture was stirred at 70 °C for 72 h. The reaction mixture was concentrated in vacuo, and the residue was partitioned between water (1000 mL) and EtOAc (800 mL). The aqueous layer was further extracted with EtOAc (500 mL × 3). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by gradient column chromatography (normal phase, silica) eluting with 0% to 15% EtOAc in hexane to give tert-butyl 2-(4-bromophenethyl)hydrazine-1-carboxylate (5.0 g, 42%) as an off-white solid. LCMS (Method 2) (m / z 260 (ES+, M+H-tBu) at 2.34 min) confirmed the product.

[0229] Step (ii): tert-Butyl 2-(4-bromophenethyl)hydrazine-1-carboxylate (5.00 g, 15.89 mmol) was dissolved in DMF (50 mL) at room temperature, potassium carbonate (10.98 g, 79.47 mmol) was added, and the mixture was stirred at room temperature for 15 minutes. 3-Bromopropionyl chloride (2.40 mL, 23.56 mmol) was then added dropwise at room temperature, and the reaction mixture was stirred at room temperature for 24 hours. The reaction mixture was concentrated in vacuo. The resulting residue was partitioned between water (500 mL) and EtOAc (400 mL), and the aqueous layer was further extracted with EtOAc (2 × 300 mL). The organic layers were combined, dried (NaSO), and the solvent was removed in vacuo. The crude product was purified by gradient column chromatography (normal phase, silica) using 0% to 30% EtOAc in hexane to give tert-butyl 2-(4-bromophenethyl)-3-oxopyrazolidine-1-carboxylate (4.0 g, 68%) as a yellow sticky solid. LCMS (Method 2) (m / z 313 (ES+, M+H-tBu) at 2.65 min) confirmed the product.

[0230] Step (iii): tert-Butyl 2-(4-bromophenethyl)-3-oxopyrazolidine-1-carboxylate (2.00 g, 5.43 mmol) was dissolved in DMF (20 mL). Nitrogen gas was purged at room temperature for 15 minutes. After this time, zinc cyanide (1.28 g, 10.87 mmol) and tetrakistriphenylphosphinepalladium 0 (0.63 g, 0.54 mmol) were added, and the reaction mixture was stirred in a microwave oven at 150 °C for 1 hour (the reaction was divided into four reaction vials). The reaction mixture was partitioned between water (400 mL) and EtOAc (300 mL). The aqueous layer was further extracted with EtOAc (2 × 200 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) eluting with 0% to 68% MeCN in water to give 4-(2-(5-oxopyrazolidin-1-yl)ethyl)benzonitrile (0.65 g, 41%) as a yellow solid. LCMS (Method 2) (m / z 216 (ES+, M+H) at 1.63 min) confirmed the product.

[0231] Step (iv): 4-(2-(5-oxopyrazolidin-1-yl)ethyl) (0.60 g, 2.79 mmol) and 1-(3-bromophenyl)prop-2-en-1-one (2.90 g, 13.95 mmol) were suspended in MeOH (6 mL) at room temperature, and the reaction mixture was stirred at room temperature for 15 minutes. After this time, TEA (2.00 mL, 13.95 mmol) was added at room temperature and stirred at 60° C. for 4 hours. The reaction mixture was partitioned between water (200 mL) and EtOAc (150 mL), and the aqueous layer was further extracted with EtOAc (120 mL×2). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) eluting the product with 0% to 72% ACN in water to give 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzoate (0.50 g, 49%) as a yellow oil. LCMS (Method 2) (m / z 426 (ES+, M+H) at 2.58 min) confirmed the product.

[0232] Step (v): 4-(2-(2-(3-(3-bromophenyl)-3-oxopropyl)-5-oxopyrazolidin-1-yl)ethyl)benzonitrile (0.50 g, 1.18 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (0.46 g, 1.76 mmol), and K2CO3 (0.33 g, 2.35 mmol) were dissolved in 1,4-dioxane (5 mL) and water (5 mL). Nitrogen gas was purged at room temperature for 20 minutes. After this time, PdCl2(dppf)·DCM (0.096 g, 0.12 mmol) was added, and the reaction mixture was stirred at 80 °C for 2 hours. The reaction mixture was partitioned between water (200 mL) and EtOAc (150 mL). The aqueous layer was further extracted with EtOAc (2 × 100 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo and the crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) eluting with 0% to 55% MeCN in water to give 3'-(3-(2-(4-cyanophenethyl)-3-oxopyrazolidin-1-yl)propanoyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (0.25 g, 44%) as an off-white solid. LCMS (Method 2) (m / z 481 (ES+, M+H) at 2.18 min) confirmed the product.

[0233] Step (vi) 3'-(3-(2-(4-cyanophenethyl)-3-oxopyrazolidin-1-yl)propanoyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (0.20 g, 0.42 mmol) was dissolved in ethanol (2 mL) and water (2 mL). After this, NaBH4 (0.03 g, 0.83 mmol) was added at 0 °C and stirred at room temperature for 1 h. The reaction mixture was partitioned between water (50 mL) and EtOAc (50 mL), and the aqueous layer was further extracted with EtOAc (2 x 30 mL). The organic layers were combined and dried (Na2SO4). The solvent was removed in vacuo to give pure 3'-(3-(2-(4-cyanophenethyl)-3-oxopyrazolidin-1-yl)-1-hydroxypropyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (0.17 g, 85%) as an off-white solid. LCMS (Method 2) (m / z 483 (ES+, M+H) at 2.14 min) confirmed the product.

[0234] Step (vii): 3'-(3-(2-(4-cyanophenethyl)-3-oxopyrazolidin-1-yl)-1-hydroxypropyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (0.17 g, 0.35 mmol) was suspended in DMF (2 mL) at room temperature, and sodium azide (0.24 g, 3.50 mmol) and ammonium chloride (0.20 g, 3.50 mmol) were added and stirred at 100°C for 48 h. The reaction mixture was partitioned between water (70 mL) and EtOAc (50 mL), and the aqueous layer was further extracted with EtOAc (3 x 50 mL). The organic layers were combined and dried (NaSO). The solvent was removed in vacuo and the crude product was purified by reverse-phase gradient flash column chromatography (reverse-phase, C18 silica) eluting the product with 0% to 45% MeCN in water to give (3'-(3-(2-(4-(1H-tetrazol-5-yl)phenethyl)-3-oxopyrazolidin-1-yl)-1-hydroxypropyl)-2-methyl-[1,1'-biphenyl]-4-carboxamide (0.10 g, 54%) as an off-white solid. LCMS (Method 2) (m / z 526 (ES+) at 1.91 min) confirmed the product. Chiral HPLC: 13.67 min and 15.68 min confirmed the purity of the product. 1 H NMR(400MHz,DMSO):1.85-1.83(d,2H,J=6.4Hz),2.29(s,3H),2.89-2.86(m,4H),3.18(S,4H),4.76(S,1H),5.40(m,1H), 7.29-7.24(m,2H),7.45-7.36(m,6H),7.76-7.76(d,1H,J=1.2Hz),7.83(s,1H),7.95-7.93(d,2H,J=8.0Hz),8.00(s,1H).

[0235] Compounds 9R and 9S: Racemic compound 9 (90 mg) was enantiomerically separated on a Waters 600 Controller-HPLC system (CHIRALPAK_IG_SFC_21 mm × 250 mm, 5 μm column) using a mobile phase of heptane (90%) and IPA and acetonitrile (70:30) (10%) without gradient and at a flow rate of 23.00 mL / min. Isomer 1: (0.028 g, 31%, white solid). LCMS (Method 2) confirmed the product (m / z 526 (ES+, M+H) at 1.75 min). Chiral HPLC confirmed the purity of the product at 13.19 min. 1 H NMR: (400 MHz, DMSO) 1.82-1.81 (d, 2H, J = 5.6 Hz), 2.10 (s, 3H), 2.87-2.83 (t, 4H, J = 6.6 Hz), 3.15 (s, 4H), 4.73 (s, 1H), 5.36 (s, 1H), 7.26-7.22 (m, 2H), 7.42-7.33 (m, 6H), 7.75-7.73 (m, 1H), 7.81 (s, 1H), 7.92-7.90 (d, 2H, J = 8.0 Hz), 7.97 (s, 1H). Isomer 2: (0.025 g, 28%, white solid). LCMS (Method 2) (m / z 526 at 1.76 min (ES+, M+H)) confirmed the product. Chiral HPLC: The purity of the product was confirmed at 14.89 min (98%). 1 H NMR:(400MHz,DMSO):1.82-1.81(d,2H,J=6.0Hz),2.26(s,3H),2.83-2.66(m,4H),3.07(s,4H),4.73(s,1H) ,5.37(s,1H),7.43-7.22(m,8H),7.75-7.73(m,1H),7.80(s,1H),7.90-7.89(d,2H,J=8.0Hz),7.98(s,1H).

[0236] Example 10: Biological Assays

[0237] Cloning, baculovirus generation, large-scale infection of HEK293 cells, and membrane preparation: Human prostaglandin E2 receptor 4 (EP4) was cloned into the pBacMam expression vector (GeneScript, UK). EP4 DNA transfer was performed using Invitrogen's Bac-to-Bac baculovirus expression system. P0 baculovirus was generated by transfecting SF9 cells with bacmid DNA using Cellfectin II transfection reagent (ThermoFisher Scientific, UK). After P0 generation, P1 virus was generated for large-scale infection and membrane preparation. HEK293 cells were grown in DMEM (ThermoFisher Scientific, UK) supplemented with 10% heat-inactivated fetal bovine serum (FBS). 5% v / v EP4 Bacman was used for 500 cm 3 Cells were infected in flasks at a seeding density of 3.5 million cells / mL. Expression was carried out for 36 hours at 37°C with 5% CO2. Cells were removed using PBS and a cell scraper. The cell culture was centrifuged at 2500 RPM for 10 minutes at 4°C. The supernatant was then poured off, 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 poured off and resuspended in 15 mL of homogenization buffer. It was 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. The resulting membranes were then stored at -80°C.

[0238] cAMP Gs functional assay: cAMP production after EP4 receptor activation was measured using a homogeneous time-resolved fluorescence (HTRF) cAMP Dynamic 2 assay (Cisbio, France). HEK293 cells were transfected with 0.5% EP4 Bacmam virus for 36 hours, after which the cells were dissociated and frozen at -150°C.

[0239] On the test day, increasing concentrations of test compound, along with a positive control (10 μM PGE2 (Tocris, Abingdon, UK)) and a negative control (DMSO (Sigma-Aldrich, UK)), were added to a ProxiPlate-384Plus white 384 shallow well microplate (PerkinElmer, USA) using an ECHO dispenser.

[0240] Cells were thawed in a water bath, resuspended in DMEM supplemented with 10% FBS, and then centrifuged at 1200 RPM for 5 minutes to form a pellet. The pellet was resuspended in assay buffer (DMEM + 0.5 mM IBMX (Tocris, Abingdon, UK)) and diluted to 0.5 × 10 6 The cell suspension (final assay concentration 5000 cells / well) was added to pre-dispensed assay plates using a multidrop. Plates were then incubated at 37°C with 5% CO for 30 minutes. cAMP production was measured according to the manufacturer's instructions, and plates were then read on a PheraStar fluorescent plate reader (BMGLabTech, Germany).

[0241] Using Dotmatics, pEC values were calculated from the midpoint of the curve as shown in Table 2. 50 The value (-LogM) was calculated.

[0242] [Table 2]

[0243] Unidirectional CACO-2 cell permeability assay Caco-2 cells (ECACC) were plated in a 24-well Transwell plate at 2 × 10 cells per well. 5Cells were seeded at 10 μM and cultured at 37°C under 5% CO for 21 days before use in confluent monolayers. Test compounds were incubated at 10 μM in assay buffer (Hanks balanced salt solution supplemented with 25 mM HEPES, adjusted to pH 6.5) (final 0.2% DMSO, n=2). Hanks balanced salt solution supplemented with 25 mM HEPES, adjusted to pH 7.4 (final 0.2% DMSO) was used in the basolateral chamber (as the receiver).

[0244] Incubations were carried out at 37°C and samples were removed from both the donor and acceptor chambers at T = 0 and 1 hour and compounds were analyzed by mass spectrometry (LC-MS / MS) including an analytical internal standard (0.5 μM carbamazepine).

[0245] Apparent transmission (P app ) values are shown in Table 3 and were determined from the following relationship:

[0246]

number

[0247] where V is the volume of each Transwell compartment (125 μL apical, 600 μL basolateral), Concentration is the relative MS response (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 is the area of the cell exposed to drug transfer (0.33 cm 2 )

[0248] To assess cell layer viability, Lucifer Yellow (LY) was added to the apical buffer in all wells. Because LY cannot freely penetrate lipophilic barriers, a high degree of LY transport indicates poor cell layer integrity, and LY P app >10×10 -6 cm / s wells were rejected.

[0249] Compound recovery from the wells was determined from the MS response (normalized to an internal standard) in the donor and acceptor chambers at the end of incubation compared to the response in the donor chamber before incubation.

[0250] [Table 3]

[0251] While the present invention has been described in conjunction with its detailed description, it should be understood that the foregoing description is intended to illustrate, but not limit, 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.

[0252] Numbered Embodiments 1. Compounds of Formula I: [ka] or a pharmaceutically acceptable salt or tautomer thereof [In the formula, 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; X is halo, OR', COOR', or C 1~6 is alkyl, L and L' are each independently C 2~4 is alkylene, R 1 H, Halo, CN, NO2, OR', SR', COOR', C 1~6 Alkoxy or C 1~6 is alkyl, R 2 , OR', OC(O)R 3 ,OC(O)OR 3 , CO2R', CON(R')2, SO2N(R')2, SO2R 3, OSO2R 3 or OSO2N(R')2, R 3 is C 1~6 Alkyl, C 3~6 cycloalkyl, or phenyl; R' is H, C 1~6 Alkyl or C 3~6 is cycloalkyl, n is 0, 1, 2, or 3; In each instance, alkyl, alkylene, alkoxy, and cycloalkyl are each optionally and independently selected from OH, SH, CN, NO, COOH, halo, or COOC. 1~4 substituted with up to three of alkyl, In each instance, heterocycloalkyl, aryl, and heteroaryl are each optionally and independently selected from OR′, SR′, CN, NO 2 , CO 2 R′, halo, C 1~4 substituted with up to three of the following: alkyl, or oxo. 2. The compound is a compound of formula (1): [ka] or a pharmaceutically acceptable salt or tautomer thereof, wherein A, X, R 1 , R 2 and n is as defined in embodiment 1. 3.A is C(O)OR',C(O)N(R')S(O)2R 3 ,S(O)2OR',C 1~8 alkyl, heterocycloalkyl, or heteroaryl, wherein said heterocycloalkyl and heteroaryl are each optionally and independently selected from OR′, SR′, halo, C 1~4 The compound of embodiment 1 or embodiment 2, wherein the compound is substituted with up to three of the following: alkyl, or oxo. 4.A is [ka] The compound of embodiment 1 or embodiment 2, selected from the group consisting of: 5.A is [ka] The compound of any one of embodiments 1 to 4, wherein 6. The compound of any one of embodiments 1-5, wherein X is halo or OR'. 7. The compound of embodiment 6, wherein X is F or OH and n is 1 or 2. 8.R 1 is optionally substituted with H, OH, halo, CN, 1 to 3 fluorine atoms 1~6 alkyl, or C optionally substituted with 1 to 3 fluorine atoms 1~6 The compound of any one of embodiments 1-7, wherein is alkyl. 9.R 1 is methyl. 10.R 2 However, CON(R')2, SO2N(R')2, OSO2R 3 or OSO2N(R')2. 11.R 2 is CONH2, SO2NH2, or OSO2NH2. 12. A compound of formula (2a), (2b), (2c), or (2d): [ka] or a pharmaceutically acceptable salt or tautomer thereof, wherein X, R 1 , R 2 and n is as defined in embodiment 1. 13. Compounds of formula (5), (5a), (5b), (6), (6a), (6b): [ka] or a pharmaceutically acceptable salt thereof. 14. The compound is selected from the group consisting of: [ka] or a pharmaceutically acceptable salt thereof. 15. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 14 and a pharmaceutically acceptable excipient. 16. A compound according to any one of embodiments 1 to 14 or a composition according to embodiment 15 for use in the treatment of an EP4 receptor-mediated disease. 17. The compound or composition for use according to embodiment 16, wherein said EP4 receptor mediated disease is a gastrointestinal disorder. 18. The compound or composition for use according to embodiment 17, wherein the gastrointestinal disorder is constipation disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, post-operative ileus, food allergy or food intolerance, celiac disease, gastrointestinal motility disorders, functional gastrointestinal disorders, drug-induced bowel diseases, NSAID-induced gastrointestinal injury, chemotherapy-induced mucositis, gastroesophageal reflux disease (GERD), duodenogastric reflux disease, diarrheal disorders, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis. 19. The compound or composition for use according to embodiment 16, wherein said EP4 receptor-mediated disease is a pulmonary disease or condition. 20. The compound or composition for use according to embodiment 19, wherein the pulmonary disease or condition is selected from chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperreactive airways disorder, and idiopathic pulmonary fibrosis.

Claims

1. Compounds of Formula I: 【Chemical 1】 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof [In the formula, A is OR', C(O)R', CO ２ R', C(O)N(R') ２ ,C(O)N(R')S(O) ２ R ３ , S(O) ２ R', S(O) ２ OR', SO ２ N(R') ２ , C １～８ alkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; X is halo, OR', COOR', or C １～６ is alkyl, L and L′ are each independently C ２～４ is alkylene, R １ H, halo, CN, NO ２ , OR', SR', COOR', C １～６ Alkoxy, or C １～６ is alkyl, R ２ is OR', OC(O)R ３ , OC(O)OR ３ , CO ２ R', CON(R') ２ , S.O. ２ N(R') ２ , S.O. ２ R ３ , OSO ２ R ３ , or OSO ２ N(R') ２ and R ３ is C １～６ Alkyl, C ３～６ cycloalkyl, or phenyl; R' is H, C １～６ Alkyl, or C ３～６ is cycloalkyl, n is 0, 1, 2, or 3; In each instance, alkyl, alkylene, alkoxy, and cycloalkyl are each optionally and independently selected from OH, SH, CN, NO ２ , COOH, halo, or COOC １～４ substituted with up to three of alkyl, In each instance, heterocycloalkyl, aryl, and heteroaryl are each optionally and independently selected from OR′, SR′, CN, NO ２ , CO ２ R', Halo, C １～４ substituted with up to three of alkyl, or oxo.

2. (i) L' is a group 【Chemistry 2】 and / or (ii) L is a group 【Chemistry 3】 2. The compound of claim 1, wherein:

3. The compound is a compound of formula (1): 【Chemistry 4】 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof, wherein A, X, R １ , R ２ 3. The compound according to claim 1 or 2, wherein n is as defined in claim 1.

4. A is C(O)OR', C(O)N(R')S(O) ２ R ３ , S(O) ２ OR', C １～８ alkyl, heterocycloalkyl, or heteroaryl, wherein said heterocycloalkyl and heteroaryl are each optionally and independently selected from OR′, SR′, halo, C １～４ The compound of any one of claims 1 to 3, substituted with up to three of alkyl or oxo.

5. A is, 【Chemistry 5】 and preferably A is selected from the group consisting of: 【Chemistry 6】 and more preferably, A is 【Chemistry 7】 The compound according to any one of claims 1 to 4,

6. A compound according to any one of claims 1 to 5, wherein X is halo or OR', preferably X is F or OH.

7. 7. The compound of claim 6, wherein X is F or OH and n is 1 or 2.

8. The compound according to any one of claims 1 to 5, wherein n is 0.

9. R １ is optionally substituted with H, OH, halo, CN, 1 to 3 fluorine atoms １～６ C optionally substituted with alkoxy or 1 to 3 fluorine atoms １～６ alkyl, preferably R １ C optionally substituted with 1 to 3 fluorine atoms １～６ The compound of any one of claims 1 to 8, which is alkyl.

10. R １ The compound of claim 9 , wherein is methyl.

11. R ２ is OR', CON(R') ２ , S.O. ２ N(R') ２ , or OSO ２ N(R') ２ and preferably, R ２ But, OH, CONH ２ , S.O. ２ NH ２ , or OSO ２ NH ２ The compound according to any one of claims 1 to 10,

12. R ２ CON(R') ２ , S.O. ２ N(R') ２ , OSO ２ R ３ , or OSO ２ N(R') ２ and preferably, R ２ But CONH ２ , S.O. ２ NH ２ , or OSO ２ NH ２ The compound according to any one of claims 1 to 10,

13. A compound of formula (2a), (2b), (2c), or (2d): 【Chemistry 8】 or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof, wherein X, R １ , R ２ and n is as defined in any one of claims 1 to 12.

14. Compounds of formula (5), (5a), (5b), (6), (6a), (6b): 【Chemistry 9】 or a pharmaceutically acceptable salt, solvate, hydrate, or tautomer thereof.

15. The compound is selected from the group consisting of: 【Chemistry 10】 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer thereof.

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15, a pharmaceutically acceptable salt, solvate, hydrate, or tautomer, and a pharmaceutically acceptable excipient.

17. 17. The pharmaceutical composition of claim 16, further comprising at least one additional therapeutic agent selected from the group consisting of an aminosalicylates, corticosteroids, immunomodulators, and combinations thereof.

18. 16. A kit comprising the compound of any one of claims 1 to 15, a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or optical isomer, and at least one additional therapeutic agent selected from the group consisting of an aminosalicylates, corticosteroids, immunomodulators, and combinations thereof.

19. A compound according to any one of claims 1 to 15, a composition according to claim 16 or 17 or a kit according to claim 18 for use as a medicament.

20. A compound according to any one of claims 1 to 15, a composition according to claim 16 or 17 or a kit according to claim 18 for use in the treatment of an EP4 receptor mediated disease.

21. 21. The compound, composition, or kit for use according to claim 20, wherein the EP4 receptor-mediated disease is a gastrointestinal disorder.

22. 22. The compound, composition, or kit for use according to claim 21, wherein the gastrointestinal disorder is constipation disorders, constipation-predominant irritable bowel syndrome, mixed irritable bowel syndrome, chronic idiopathic constipation, gastrointestinal symptoms associated with Parkinson's disease, gastrointestinal symptoms associated with cystic fibrosis, intestinal motility disorders, post-operative ileus, food allergy or food 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 disease, diarrheal disorders, immune-mediated gastrointestinal diseases, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and ischemic colitis.

23. 21. The compound, composition, or kit for use according to claim 20, wherein the EP4 receptor-mediated disease is a pulmonary disease or condition.

24. 24. The compound, composition, or kit for use according to claim 23, wherein the pulmonary disease or condition is selected from chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hyperreactive airways disorder, and idiopathic pulmonary fibrosis.

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