GPR35 agonist compounds

Abstract

The present disclosure relates to novel compounds of formula (1) [Formula 1] TIFF2024517132000097.tif30144 and its salts and any tautomers thereof (wherein X, R 1 and R 2 as described herein) and their use for treating, preventing, ameliorating, controlling, or reducing the risk of disorders associated with the GPR35 receptor.

Description

[Technical Field] 【0001】 This application relates to compounds and their use as G protein-coupled receptor 35 (GPR35) receptor agonists. The compounds described herein may be useful in treating or preventing diseases involving the GPR35 receptor. [Background technology] 【0002】 GPR35 is an orphan receptor belonging to the family of seven-transmembrane domain G protein-coupled receptors (GPCRs). The GPCR superfamily represents a large family of signaling molecules that play a crucial role in regulating various aspects of human physiology. Due to their pharmacological responsiveness, these receptors have been intensively studied as potential drug targets. Recent analyses have shown that more than 475 drugs act on 108 intrinsic GPCRs, representing approximately 34% of FDA-approved drugs. Opportunities for the discovery and development of novel drugs for orphan GPCRs whose endogenous ligands have yet to be identified remain. 【0003】 GPR35 was initially discovered as an open reading frame encoding a 309-amino acid protein and is localized to human chromosome 2q37.3 (O'Dowd et al. Genomics, 47, 310-3, 1998). The receptor has been shown to be expressed at high levels within a range of tissues, and has been reported in gastrointestinal tissue, lung, and dorsal root ganglia. The receptor has also been found to be expressed in immune cells and in tissues, such as the spleen, skeletal muscle, and spinal cord. 【0004】 In line with its expression pattern, there is growing evidence highlighting the therapeutic role of GPR35 across a wide range of conditions, including respiratory diseases, metabolic syndrome (e.g., diabetes), cardiovascular diseases (e.g., hypertension), and pain. Ligands targeting GPR35 may offer potential benefits for treating a broad range of human disease conditions. 【0005】 The identity of the endogenous ligand for GPR35 remains a subject of debate, with several putative ligands, including 2-acyllysophosphatidic acid, CXCL17, and kynurenic acid, being expressed to date. Many of the reported ligands exhibit weak activity at the receptor or lack biological specificity, raising questions about the true identity of the endogenous ligand. 【0006】 A broad range of synthetic agonists have been reported to act with GPR35, including zaprinast, pamoic acid, cromolyn, loop diuretics (bumenide, furosemide), aspirin metabolites, quercetin, and dicumarol. In addition, weak agonist activity has also been reported with the anti-inflammatory agents sulfasalazine and 5-aminosalicylic acid, which are widely used in the treatment of inflammatory bowel disease (EC50 of approximately 3 μM) (U.S. Patent Application Publication US2013 / 0316985). 【0007】 Tylphostine (Tylphostine-51), a tyrosine kinase class compound, and the catechol-O-methyltransferase (COMT) inhibitor, entacapone, have also been reported to act on receptors that highlight the diversity of the GPR35 ligand class. Due to the potential involvement of GPR35 in human diseases, there is growing interest in developing highly effective ligands that exhibit selectivity for GPR35. The elucidation of GPR35 biology has been somewhat hampered by the lack of sufficient pharmacological tools. Indeed, many of the identified GPR35 agonists (endogenous and synthetic) exhibit only weak or partial activity on GPR35, lack target specificity, and make dissection of the pathway difficult. Furthermore, some compounds have been shown to exhibit species selectivity and, in some cases, ligand bias. The putative endogenous ligand kynurenic acid is one such example, reported to be at least 100 times less potent in the human receptor compared to its rat ortholog (Jenkins et al. Br J Pharmacol 162, 733-748, 2011). Therefore, potent and selective GPR35 agonists and antagonists are needed to elucidate the physiological role of this receptor. 【0008】 More recently, genome-wide association studies (GWAS) have identified single nucleotide polymorphisms (SNPs) of GPR35 associated with inflammatory bowel disease (IBD). Two SNPs are expressed, one of which is a non-synonymous SNP (rs3749171) encoding a T108M substitution in a third transmembrane domain (Ellinghaus et al. Hepatology 58, 1074-1083, 2013). This residue is not conserved across mammals, and the impact of this polymorphism on signaling remains unclear. The second polymorphism at the GPR35 locus (rs4676410) encodes an upstream intron variant of GPR35. A phenom-wide association analysis using four large real-world data cohorts demonstrated an association between this genetic variant (rs4676410) and IBD phenotypes that support previously reported target-disease links (Diogo et al. Nat Commun 9, 4285, 2018). The reported association of GPR35 polymorphisms in IBD is increasing interest in the potential therapeutic role of GPR35 for the treatment of gastrointestinal diseases. 【0009】 Inflammatory bowel disease (IBD) is a chronic, recurrent inflammatory gastrointestinal disorder that typically involves the ileum and / or colon. Disease physiology suggests that an abnormal immune response in the intestines is involved, leading to mucosal inflammation, a defective intestinal barrier, and increased glycemic index (GI) permeability. Treatment strategies largely involve a stepwise approach through the combined use of chemicals, such as aminosalicylates, corticosteroids, immunosuppressants, biologics (e.g., anti-TNF), and antibiotics. However, it is emphasized that many patients do not achieve complete disease control, and this is highly anticipated. One particularly under-treated and under-recognized characteristic is abdominal pain. Pain is a common symptom experienced by the majority of IBD patients during the course of the disease and can be a direct or indirect consequence of intestinal inflammation. Whatever the cause, pain negatively impacts the quality of life for IBD patients. To date, managing IBD-related abdominal pain remains a challenge. Commonly used analgesics, such as non-steroidal anti-inflammatory drugs (NSAIDs), often worsen symptoms and leave patients with limited treatment options. Therefore, there is a great need for chemicals that can provide rapid relief of pain onset, and the development of new drugs is urgently required. 【0010】 In preclinical studies, GPR35 mutant mice exhibited more severe colonic epithelial damage after chemical injury compared to wild-type mice. GPR35 knockout mice showed increased inflammatory expression and remodeling cytokines, but the number of inflammatory cells influx into the mucosa did not show an overall difference (Farooq et al. Digestive Diseases and Sciences, 63, 2910-22, 2018). The role of GRP35 in barrier homeostasis has also been reported, along with chemicals such as sodium cromoglycate, which has shown the ability to reduce GI permeability in several intestinal sensitization models (Forbes et al. J Exp Med 205: 897-913, 2008; Yokooji et al. Int Arch Allergy Immunol. 167:193-202, 2015). Consistent with these findings, in vitro studies have shown that GPR35 plays a role in restricting tight junction proteins and promoting epithelial cell migration. Ultimately, GPR35 is abundantly expressed in dorsal root ganglion (DRG) neurons, where it has been shown to co-localize with nociceptive ion channels and play a role in the pain process (Ohshiro et al. Biochem. Biophys. Res. Commun. 365, 344-8, 2008). In addition to its reported effects on barrier defense, chromolyne reduces visceral hypersensitivity in stress-sensitive rat strains (Carroll et al. PLoS One.8:e84718, 2013), highlighting its potential usefulness in pain management. 【0011】 Sodium cromoglycate is a mast cell stabilizer used in the prophylaxis of systemic mast cell disease, allergic rhinitis and asthma, and in the manifestations of allergic conjunctivitis and (in conjunction with dietary restrictions) food allergies. In systemic mast cell disease, the use of cromoglycates has been reported to improve diarrhea, facial flushing, headache, vomiting, urticaria, and abdominal pain. Trials evaluating the effects of sodium cromoglycate in food allergies have reported mixed results, and high doses are usually required to provide protection. A maximum dose of 2 g / day has been shown to be effective in reducing the severity of GI symptoms in patients with irritable bowel syndrome caused by food allergies (Lunardi et al. Clin Exp Allergy. 21:569-72, 1991). Similar findings have been reported in children with milk allergies at the gastrointestinal permeability endpoint. 【0012】 In addition to gastrointestinal disorders, GPR35 has attracted interest as a target for the treatment of allergic disorders, including asthma. In the lungs, cromolyn has long been used as an effective asthma therapy with a good safety and tolerable profile, but it is associated with suboptimal pharmacokinetics. It is estimated that approximately 5–12% of the drug is absorbed after airway deposition, and more recently, improved formulations of cromolyn that achieve significantly higher drug deposition in the lungs have been developed (PA101). 【0013】 Clinically, chromolyne demonstrates efficacy in suppressing immediate and delayed asthmatic responses following allergen challenge. In a Phase II proof-of-concept trial, PA101 demonstrated efficacy in reducing cough frequency in patients with idiopathic pulmonary fibrosis. GPR35 mRNA is upregulated in response to IgE antibody-mediated challenge, and chromolyne has been reported to inhibit inflammatory mediator release in human lung flakes passively sensitized with IgE antibodies. These effects appear to involve multiple mechanisms, including mast cell stabilization and limitation of reflex-induced bronchoconstriction. 【0014】 Therefore, the new data highlight the broad therapeutic potential of GPR35 agonists, extending to the treatment of mast cell damage, acute and chronic pain conditions, and diseases associated with allergic or inflammatory disorders in both the gastrointestinal tract and the lungs. [Overview of the project] [Problems that the invention aims to solve] 【0015】 The present invention relates to a compound having activity as a G protein-coupled receptor 35 (GPR35) receptor agonist. [Means for solving the problem] 【0016】 In short, in one embodiment, the present invention relates to a compound of formula (1): 【0017】 [ka] or its salt or its tautomer (In the formula, X is N or CH, R 1 H or halo, R 2 H, halo, and C are substituted in some cases. 1~6 Alkyl, and in some cases substituted C 3~6 Cycloalkyl, and in some cases substituted C 1~6 The present invention provides an alkoxy, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted O-aryl. 【0018】 The compound can be used as a GPR35 receptor agonist. The compound can be used in the manufacture of pharmaceuticals. The compound may be used to treat, prevent, improve, control, or reduce the risk of GPR35-related disorders. The compound can be used to treat mast cell damage, acute and chronic pain conditions, and diseases associated with allergic or inflammatory diseases of both the gastrointestinal tract and the lungs. [Brief explanation of the drawing] 【0019】 [Figure 1] This is the X-ray powder diffraction pattern of the crystalline form of the tromethamine salt (hydrate I) of compound A. [Figure 2] This is a differential scanning calorimeter curve of the crystalline form of the tromethamine salt (hydrate I) of compound A. [Figure 3] This is the thermogravimetric analysis curve of the crystalline form of the tromethamine salt (hydrate I) of compound A. [Figure 4] This is the X-ray powder diffraction pattern of the crystalline form of tromethamine salt (hydrate II). [Figure 5] This is a differential scanning calorimeter curve of the crystalline form of the tromethamine salt (hydrate II) of compound A. [Figure 6] This is the thermogravimetric analysis curve of the crystalline form of the tromethamine salt (hydrate II) of compound A. [Figure 7] This is the X-ray powder diffraction pattern of the crystalline form of the free acid of compound A (pattern 3). [Figure 8] This is the thermogravimetric analysis curve of the crystalline form of the free acid (pattern 3) of compound A. [Figure 9] This is the X-ray powder diffraction pattern of the crystalline form of the free acid of compound A (pattern 1). [Figure 10] This is the thermogravimetric analysis curve of the crystalline form of the free acid (pattern 1) of compound A. [Modes for carrying out the invention] 【0020】 The present invention relates to novel compounds. The present invention relates to the use of novel compounds as agonists of the GPR35 receptor. The present invention also relates to the use of novel compounds in the treatment or prevention of diseases involving the GPR35 receptor. The present invention further relates to the use of novel compounds in the manufacture of pharmaceuticals for use as GPR35 receptor agonists. 【0021】 The present invention relates to a compound of formula (1) 【0022】 【Chemical formula】 or a salt thereof or a tautomer thereof (wherein X is N or CH, R 1 is H or halo, R 2 is H, halo, optionally substituted C 1~6 alkyl, optionally substituted C 3~6 cycloalkyl, optionally substituted C 1~6 alkoxy, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted O-aryl). 【0023】 In the compounds of the present specification, X may be N. X may be CH. 【0024】 In the compounds of the present specification, R 1 may be H. R 1 may be halo. R 1 may be Cl or F. R 1 may be Cl. R 1 may be F. R 1 may be Br. 【0025】 In the compounds of the present specification, R 2 may be H. R 2 may be halo. R 2 is optionally substituted C 1~6Alkyl may also be used. 2 C is replaced in some cases. 3~6 A cycloalkyl group may also be used. 2 C is replaced in some cases. 1~6 Alkyl compounds are also acceptable. 2 R may be a substituted aryl depending on the case. 2 R may be a heteroaryl that is substituted in some cases. 2 R may be a substituted monocyclic heteroaryl depending on the circumstances. 2 R may be a bicyclic heteroaryl that is substituted in some cases. 2 This may be an O-aryl that is substituted in some cases. 【0026】 R 2 C is substituted with H, and in some cases with one to six fluorine atoms. 1~6 Alkyl, and in some cases C substituted with one to six fluorine atoms 3~6 Cycloalkyl, or optionally substituted with one to six fluorine atoms, C 1~6 Alkyl compounds are also acceptable. 2 The compound may be H, trifluoromethyl, ethyl, cyclopropyl, cyclohexyl, or methoxy. 【0027】 R 2 R 3 Phenyl substituted with, in some cases R 3 Pyridyl is substituted with, in some cases R 3 O-phenyl substituted with, and in some cases R 3 Indazolyl substituted with, or R in some cases 3 It may also be a pyridazinyl substituted with R 3 C is substituted with H, halo, and in some cases one to six fluorine atoms. 1~6 Alkyl, and in some cases C substituted with one to six fluorine atoms 3~6 Cycloalkyl, sometimes substituted with one to six fluorine atoms, C 1~6 Alkoxy, -CO2R 4-CONHCH2R 4 -CONHCH2CH2OR 4 , -OR 4 ,-OCH2R 4 , -CH2R 4 ,-OCH2R 4 -CH2CH2OR 4 ,-OCH2CH2OR 4 , -CONHR 4 Or -CON(CH3)R 4 And R 4 C is substituted with H, and in some cases with one to six fluorine atoms. 1~6 Alkyl, or group: 【0028】 [ka] And R 5 , R 6 and R 7 These are independently H, halo, and CO2R. 8 CONR 8 R 9 or, in some cases, C substituted with one to six fluorine atoms 1~6 It is alkyl, C 1~6 One or two carbon atoms of the alkyl group may be replaced by oxygen in some cases. R 8 and R 9 C is independently substituted with H or optionally 1 to 6 fluorine atoms. 1~6 It is alkyl. 【0029】 R 2 may be a substituted aryl, a substituted heteroaryl, or a substituted O-aryl, and any substituent is R 3 That is the case. 【0030】 R 2It may be optionally substituted aryl, optionally substituted O-aryl, optionally substituted heteroaryl, optionally substituted monocyclic heteroaryl, or optionally substituted bicyclic heteroaryl, and any substituent is R 3 is as follows. 【0031】 R 2 may be phenyl optionally substituted with R 3 pyridyl optionally substituted with R 3 O-phenyl optionally substituted with R 3 indazolyl optionally substituted with R 3 or pyridazinyl optionally substituted with R 3 is also acceptable. 【0032】 R 3 is H, halo, C 1~6 alkyl optionally substituted with from 1 to 6 fluorine atoms, C 3~6 cycloalkyl optionally substituted with from 1 to 6 fluorine atoms, C 1~6 alkoxy, -CO2R 4 -CONHCH2R 4 -CONHCH2CH2OR 4 -OR 4 -OCH2R 4 -CH2R 4 -OCH2R 4 -CH2CH2OR 4 -OCH2CH2OR 4 -CONHR 4 or -CON(CH3)R 4 is also acceptable, R 4 is H, C 1~6 alkyl optionally substituted with from 1 to 6 fluorine atoms, or the group: 【0033】 【Chemical formula】 is as follows, and R 5 、R6 and R 7 are independently H, halo, CO2R 8 , CONR 8 R 9 , or optionally C substituted with from 1 to 6 fluorine atoms 1~6 alkyl, and one or two carbon atoms of the C 1~6 alkyl group may optionally be replaced by O, R 8 and R 9 are independently H or optionally C substituted with from 1 to 6 fluorine atoms 1~6 alkyl. 【0034】 In the compounds of this specification, R 4 is H, methyl, or 【0035】 【Chemical formula】 may be selected from the group consisting of. In the compounds of this specification, R 5 , R 6 and R 7 may independently be H, CF3, CONH2 or -OCH2CH2OCH3. 【0036】 In the compounds of this specification, R 8 and R 9 may independently be H or methyl. R 3 is OMe, CO2H, CO2Et, CON(CH3)2, CONHCH2CH2OCH3, or 【0037】 【Chemical formula】 may be selected from the group consisting of. 【0038】 In the compounds of this specification, R 2 is 【0039】 【Chemical formula】 The group consisting of TIFF0007875883000008.tif197145 may be selected. 【0040】 The compound is a compound of formula (1a) or (1b). 【0041】 [ka] Or it may be a salt thereof or a tautomer thereof, R 1 and R 2 This is as stipulated above. 【0042】 In some embodiments, the compound is a compound of formula (1a). 【0043】 [ka] Or it may be a salt thereof or a tautomer thereof, R 1 and R 2 This is as stipulated above. 【0044】 The compound is a compound of formula (2a) or (2b): 【0045】 [ka] Or it may be a salt thereof or a tautomer thereof, R 2 This is as stipulated above. 【0046】 In some embodiments, the compound is a compound of formula (2a). 【0047】 [ka] Or it may be a salt thereof or a tautomer thereof, R 2 This is as stipulated above. 【0048】 The compounds are those of formula (3a), (3b), (3c), (3d), or (3e): 【0049】 [ka] TIFF0007875883000014.tif93128 or its salts or tautomers, X, R 1 and R 3 This is as stipulated above. 【0050】 In some embodiments, the compound is a compound of formula (3a). 【0051】 [ka] Alternatively, it may be a salt thereof or a tautomer thereof, X, R 1 and R 3 This is as stipulated above. 【0052】 The compounds are those of formula (4a), (4b), (4c), (4d), or (4e). 【0053】 [ka] TIFF0007875883000017.tif94128 【0054】 Or it may be a salt thereof or a tautomer thereof, R 3 This is as stipulated above. 【0055】 In some embodiments, the compound is a compound of formula (4a). 【0056】 [ka] Or it may be a salt thereof or a tautomer thereof, R 3 This is as stipulated above. 【0057】 The compound is 【0058】 [ka] The following may be selected from the group consisting of TIFF0007875883000020.tif220153TIFF0007875883000021.tif196154TIFF0007875883000022.tif173154, or they may be salts thereof or tautomers thereof. 【0059】 The compound is 3-((3-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((5-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-yl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; Ethyl 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazol-5-yl)-[1,1'-biphenyl]-3-carboxylate; N-benzyl-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; N-benzyl-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-carboxamide; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(2-methoxyethyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(4-(2-methoxyethoxy)benzyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; 3-Hydroxy-4-((3'-Methoxy-5-(1H-Tetazol-5-yl)-[1,1'-Biphenyl]-3-yl)amino)cyclobuta-3-en-1,2-dione; 3-((3-ethyl-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-carboxylic acid; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-N-(4-(trifluoromethyl)benzyl)-[1,1'-biphenyl]-4-carboxamide; 3-((4-fluoro-3-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((4-chloro-3-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((5-(1H-tetrazole-5-yl)-4'-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(6-(benzyloxy)pyridine-3-yl)-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((6-Methoxy-4-(1H-Tetazol-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((6-phenyl-4-(1H-tetrazole-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazol-5-yl)-5-(2-(4-(trifluoromethyl)benzyl)-2H-indazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(6-((4-(trifluoromethyl)benzyl)oxy)pyridine-3-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(2-benzyl-2H-indazole-5-yl)-5-(1H-tetrazol-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((6-Phenoxy-4-(1H-Tetazol-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(2-(2-(4-(trifluoromethyl)phenoxy)ethyl)-2H-indazole-6-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-cyclopropyl-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-indazole-5-yl)-5-(1H-tetrazol-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(6-(2-(3,5-bis(trifluoromethyl)phenoxy)ethoxy)pyridine-3-yl)-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(6-(2-(4-(trifluoromethyl)phenoxy)ethoxy)pyridine-3-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-cyclohexyl-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((3-(pyridazin-4-yl)-5-(1H-tetrazole-5-yl)phenyl)amino)cyclobuta-3-en-1,2-dione; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N,N-dimethyl-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; N-(4-carbamoylbenzyl)-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide They may be selected from the group consisting of the above, or they may be salts thereof or tautomers thereof. 【0060】 In some embodiments, salts of the compound of formula (1) are pharmaceutically acceptable salts. 【0061】 In some embodiments, the compound has the following structure: 【0062】 [ka] A compound having, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound has the following structure: 【0063】 [ka] It is a compound that has [a certain characteristic]. 【0064】 This disclosure also has the following structure: 【0065】 [ka] The present invention provides a trometamine salt having the following properties. 【0066】 This disclosure also provides crystalline forms of compound A or pharmaceutically acceptable salts thereof. In some embodiments, the crystalline forms include the free acid of compound A or the tromethamine salt of compound A. 【0067】 In some embodiments, the crystalline form comprises the tromethamine salt of compound A. In some embodiments, the crystalline tromethamine salt of compound A is a hydrate. In one embodiment, the crystalline tromethamine salt of compound A is hydrate I. In one embodiment, the crystalline tromethamine salt of compound A is characterized by an XRPD pattern substantially according to Figure 1. In one embodiment, the crystalline tromethamine salt of compound A is characterized by an XRPD pattern with diffraction angles of 3.9±0.2, 7.7±0.2, 10.0±0.2, and 15.8±0.2°2θ when measured using CuKα radiation. 【0068】 In one embodiment, the tromethamine salt of crystalline compound A is hydrate II. In one embodiment, the tromethamine salt of crystalline compound A is characterized by an XRPD pattern substantially according to Figure 4. In one embodiment, the tromethamine salt of crystalline compound A is characterized by an XRPD pattern with diffraction angles of 4.4±0.2, 14.4±0.2, and 23.9±0.2°²θ when measured using CuKα radiation. 【0069】 In some embodiments, the crystalline form includes compound A (free acid). In some embodiments, the crystalline compound A (free acid) is a hydrate. In some embodiments, the crystalline compound A (free acid) is pattern 1. In some embodiments, the crystalline compound A (free acid) is pattern 3. 【0070】 In one embodiment, the free acid of crystalline compound A is characterized by an XRPD pattern substantially according to Figure 7. In another embodiment, the free acid of crystalline compound A is characterized by an XRPD pattern substantially according to Figure 9. 【0071】 The compounds disclosed herein may be used in therapeutic applications. The compounds disclosed herein may be used in pharmaceutical applications. 【0072】 The compound can be used as a GPR35 receptor agonist. The compound can be used in the manufacture of pharmaceuticals. The compound may be used to treat, prevent, improve, control, or reduce the risk of GPR35-related disorders. The compound can be used to treat or prevent mast cell damage, acute and chronic pain conditions, and diseases associated with allergic or inflammatory diseases of both the gastrointestinal tract and the lungs. 【0073】 The compound may be used to treat gastrointestinal disorders and diseases using a chemical that selectively acts on the GPR35 receptor. These include, but are not limited to, food allergies, food intolerances and allergic disorders, celiac disease, systemic mast cell disorders and other mast cell-related disorders (mast cell activation syndrome, clonal mast cell disorder, monoclonal mast cell activation syndrome, idiopathic urticaria, idiopathic anaphylaxis), mastocytic colitis, irritable bowel syndrome (IBS), gastrointestinal motility disorders, functional gastrointestinal disorders, gastroesophageal reflux disease (GERD), duodenal reflux, diarrheal diseases, eosinophilic gastroenteritis, eosinophilic esophagitis, infectious diarrhea (e.g., Clostridium difficile, Salmonella, Shigella toxin), microscopic colitis, immune-mediated gastrointestinal disorders, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and visceral abdominal pain. In some embodiments, the compound may be used to treat irritable bowel syndrome (IBS), including IBS with constipation (IBS-C), IBS with diarrhea (IBS-D), and IBS with mixed bowel movements (IBS-M). In some embodiments, the compound may be used to treat IBS-D. 【0074】 In some embodiments, the compound may be used to treat inflammatory bowel disease (IBD). In some embodiments, the compound may be used to treat Crohn's disease. In some embodiments, the compound may be used to treat ulcerative colitis. 【0075】 The compound may be used to treat pain symptoms associated with gastrointestinal diseases and other visceral diseases, including Crohn's disease, ulcerative colitis, inflammatory bowel disease, radiation colitis, radiation cystitis, celiac disease, glutenic enteropathy, radiation cystitis, interstitial cystitis, bladder pain syndrome, cancer, gastroesophageal reflux disease, chemotherapy mucositis and radiation therapy mucositis, pancreatitis, prostatitis, pelvic pain, endometriosis, hepatitis, hepatic fibrosis and cirrhosis. 【0076】 The compounds may be used to treat lung diseases and illnesses, including, but not limited to, chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hypersensitivity respiratory disease, and idiopathic pulmonary fibrosis. 【0077】 This application also provides methods for treatment by any use of the compound of formula (1) described herein. In some embodiments, methods are provided for treating a GPR35-related disorder in a subject requiring treatment, comprising administering a therapeutically effective amount of the compound of formula (1) (e.g., compound A) or a pharmaceutically acceptable salt thereof to the subject. In some embodiments, the subject is human. 【0078】 In some embodiments, the disorder is inflammatory bowel disease (IBD). In some embodiments, the disorder is Crohn's disease. In some embodiments, the disorder is ulcerative colitis. 【0079】 In some embodiments, the disorder is irritable bowel syndrome (IBS), including IBS with constipation (IBS-C), IBS with diarrhea (IBS-D), and IBS with mixed bowel habits (IBS-M). In some embodiments, the disorder is IBS-D. 【0080】 definition In this application, unless otherwise indicated, the following definitions apply. 【0081】 The term “treatment” in relation to the use of any compound described herein, including the compound of formula (1), is used to describe any form of intervention involving the administration of a compound to a subject who has, is at risk of, or is at risk of having, the disease or disorder in question. Thus, the term “treatment” encompasses both prophylactic (preventive) treatments and interventions that result in the manifestation of measurable or detectable symptoms of the disease or disorder. 【0082】 The term "therapeutic effective dose" (e.g., in relation to a disease or treatment method for a patient) refers to the amount of a compound that is effective in producing a desired therapeutic effect. For example, if the patient is pain, the therapeutic effective dose is the amount sufficient to provide the desired level of pain relief. The desired level of pain relief may be, for example, complete elimination of pain or a reduction in the severity of pain. 【0083】 Terms, for example, "C 1~6 The "alkyl" in "alkyl", for example, "C 3~6 The "cycloalkyl" in "cycloalkyl," for example, "C 1~6 The terms "alkoxy," "aryl," "heteroaryl," "monocyclic," and "bicyclic" in the term "alkoxy" are all used in their conventional sense (for example, as defined in the IUPAC Gold Book) unless otherwise indicated. 【0084】 To the extent that any compound described contains a chiral center, the present invention extends to all optical isomers of such compounds, whether in racemic or divided enantiomer form. The present invention as described herein relates to all crystalline forms, solvates, and hydrates of any of the disclosed compounds, although they have been prepared as described above. To the extent that any of the compounds disclosed herein contains an acidic or basic center, such as a carboxylate group or an amino group, all salt forms of such compounds are included herein. In the case of pharmaceutical use, salts should be considered pharmaceutically acceptable salts. 【0085】 The compounds of the present invention may exist in the form of tautomers. Any reference to a named compound or a compound expressed by its structure is understood to be intended to encompass all tautomers of such compound. For example, the compound of formula (1) has the following tautomers: 【0086】 [ka] It includes. 【0087】 Salts that may be mentioned or pharmaceutically acceptable salts include acid addition salts and base addition salts. Such salts can be formed by conventional methods, for example, by reacting a compound in the form of a free acid or free base with one equivalent or more of a suitable acid or base in an optional solvent or in a medium insoluble in the salt, followed by removal of the solvent or medium using standard methods (e.g., by vacuum, freeze-drying, or filtration). Salts can also be prepared by exchanging the counterion of a compound in the form of a salt with another counterion, for example, using a suitable ion exchange resin. 【0088】 Examples of pharmaceutically acceptable salts include acid addition salts derived from inorganic and organic acids, as well as salts derived from metals, such as sodium, magnesium, potassium, and calcium. Representative pharmaceutically acceptable base addition salts, though not limited to these, include aluminum, ammonium, 2-amino-2-(hydroxymethyl)-1,3-propanediol (TRIS, tromethamine), arginine, benethamine (N-benzylphenethylamine), benzathine (N,N'-dibenzylethylenediamine), bis-(2-hydroxyethyl)amine, bismuth, calcium, chloroprocaine, choline, and cremisole (1-p-chlorobenzyl-2-pyrrolidine). The present invention includes (-1'-ylmethylbenzimidazole), cyclohexylamine, dibenzylethylenediamine, diethylamine, diethyltriamine, dimethylamine, dimethylethanolamine, dopamine, ethanolamine, ethylenediamine, L-histidine, iron, isoquinoline, lepidine, lithium, lysine, magnesium, meglumine (N-methylglucamine), piperazine, piperidine, potassium, procaine, quinine, quinoline, sodium, strontium, t-butylamine, and zinc. In one embodiment, the salt of the compounds disclosed herein is a tromethamine (TRIS) salt. 【0089】 Examples of acid addition salts include acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, aryl sulfate (e.g., benzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, and p-toluenesulfonic acid), ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzoic acid, 4-acetamidobenzoic acid, butanoic acid, (+)camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfonic acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid (e.g., D-gluconic acid), glucuronic acid (e.g., D- Glucuronic acid, glutamic acid (e.g., L-glutamic acid), α-oxoglutaric acid, glycolic acid, hypric 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, metaphosphate, methanesulfonic acid, 1-hydro Examples include acid addition salts formed from xy-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, sulfonic acid, tannic acid, tartaric acid (e.g., (+)-L-tartaric acid), thiocyanic acid, undecylenic acid, and valeric acid. 【0090】 This also includes any solvates of the compounds and their salts. Preferred solvates are those formed by introducing molecules of a non-toxic, pharmaceutically acceptable solvent (hereinafter referred to as the solvating solvent) into the solid structure (e.g., crystalline structure) of the compound of the present invention. Examples of such solvents include water, alcohols (e.g., ethanol, isopropanol, and butanol), and dimethyl sulfoxides. Solvates can be prepared by recrystallizing the compound of the present invention in a solvent or mixture of solvents containing the solvating solvent. In any case, whether or not a solvate has been formed can be determined by analyzing the crystal of the compound using well-known and standard methods, such as thermomass spectrometry (TGA), differential scanning calorimeter (DSC), and X-ray crystallography. 【0091】 Solvates may be stoichiometric or non-stoichiometric solvates. Certain solvates may also be hydrates, examples of which 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, Indiana, USA, 1999, ISBN 0-967-06710-3. 【0092】 In the context of the present invention, the term "pharmaceutical composition" means a composition comprising an activator and a composition further comprising one or more pharmaceutically acceptable carriers. Depending on the nature of the method of administration and the dosage form, the composition may further contain raw materials selected from, for example, diluents, adjuvants, excipients, vehicles, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavorings, antibacterial agents, antifungal agents, lubricants, and dispersants. The composition may take the form of, for example, liquid formulations including tablets, sugar-coated tablets, powders, elixirs, syrups, suspensions, sprays, inhalants, tablets, lozenges, emulsions, liquids, cachets, granules, capsules, and suppositories, as well as liquid formulations for injection including liposomal formulations. 【0093】 The compounds of the present invention may contain one or more isotopic substitutions, and a reference to a particular element includes all isotopes of that element within that range. For example, a reference to hydrogen includes all isotopes within that range. 1 H, 2 H(D), and 3 Includes H(T). Similarly, references to carbon and oxygen are within their respective ranges. 12 C, 13 C and 14 C and 16 O and 18 Each of the O groups is included. In similar methods, references to specific functional groups also include isotopic variants within their scope unless otherwise specifically indicated in the context. For example, references to alkyl groups, e.g., ethyl groups or alkoxy groups, e.g., methoxy groups, also include variants in which one or more hydrogen atoms of the group are in the form of deuterium or tritium isotopes, such as an ethyl group in which all five hydrogen atoms are in the form of deuterium isotopes (perduteroethyl group), or a methoxy group in which all three hydrogen atoms are in the form of deuterium isotopes (triduteromethoxy group). The isotopes may be radioactive or non-radioactive. 【0094】 Therapeutic doses can be adjusted according to the patient's needs, the severity of the disease being treated, and the compound being used. Determining the appropriate dosage in specific situations is within the scope of those skilled in the art. Generally, treatment is initiated with a dose lower than the optimal dose of the compound. The dose is then gradually increased until the optimal effect under these conditions is achieved. For convenience, the total daily dose can be divided into smaller doses throughout the day, if desired. 【0095】 The effective dose of a compound naturally varies depending on the severity of the disease being treated and on the specific compound and its route of administration. Selecting an appropriate dose is within the capabilities of those skilled in the art and does not require undue burden. Generally, the daily dose range may be about 10 μg to about 30 mg per kg of body weight of humans and non-human animals, preferably about 50 μg to about 30 mg per kg of body weight of humans and non-human animals, for example, about 50 μg to about 10 mg per kg of body weight of humans and non-human animals, for example, about 100 μg to about 30 mg per kg of body weight of humans and non-human animals, for example, about 100 μg to about 10 mg per kg of body weight of humans and non-human animals, and most preferably about 100 μg to about 1 mg per kg of body weight of humans and non-human animals. 【0096】 Pharmaceutical preparations Even if it is possible to administer the active compound alone, it is preferable to provide it as a pharmaceutical composition (e.g., a formulation). 【0097】 Accordingly, in some embodiments of the present invention, a pharmaceutical composition is provided comprising at least one compound of formula (1) or a salt thereof, as defined above, together with at least one pharmaceutically acceptable excipient. 【0098】 Pharmaceutically acceptable excipients can be selected from, for example, carriers (e.g., solid, liquid, or semi-solid carriers), adjuvants, diluents (e.g., solid diluents, e.g., fillers or bulking agents, and liquid diluents, e.g., solvents and co-solvents), granulators, binders, flow aids, coatings, release control agents (e.g., release-delay polymers or release-delay waxes), binders, tablet decomposition agents, buffers, lubricants, preservatives, antifungal and antimicrobial agents, antioxidants, buffers, isotonic modifiers, consistency enhancers, flavorings, sweeteners, pigments, plasticizers, taste masking agents, stabilizers, or any other excipients conventionally used in pharmaceutical compositions. 【0099】 As used herein, the term “pharmaceutically acceptable” means a compound, material, composition, and / or dosage form suitable for use in contact with the tissue of a subject (e.g., a human subject) within the bounds of sound medical judgment and in proportion to a reasonable risk-benefit ratio, without excessive toxicity, irritation, allergic reaction, or other problems or complications. Each excipient must also be “acceptable” in the sense that it is compatible with the other ingredients of the formulation. 【0100】 Pharmaceutical compositions containing the compound of formula (1) can be formulated according to known techniques, see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, United States. The pharmaceutical compositions may be in any form suitable for oral, parenteral, intravenous, intramuscular, intravenous, intramuscular, intrathecal, subcutaneous, topical, intranasal, intrabronchial, sublingual, oral cavity, ophthalmic, otological, rectal, vaginal, or transdermal administration. 【0101】 Suitable dosage forms for orally administered pharmaceuticals include tablets (coated or uncoated), capsules (hard shell or soft shell), caplets, pills, lozenges, syrups, liquids, powders, granules, elixirs and suspensions, sublingual tablets, wafers or patches, such as oral patches. 【0102】 The composition may be a tablet composition or a capsule composition. The tablet composition may contain a unit dose of the active compound together with an inert diluent or carrier, such as sugars or sugar alcohols, such as lactose, sucrose, sorbitol, or mannitol, and / or a non-sugar diluent, such as sodium carbonate, calcium phosphate, calcium carbonate, or cellulose or its derivatives, such as microcrystalline cellulose (MCC), methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, and starch, such as corn starch. The tablets may also contain standard raw materials, such as binders and granulators, such as polyvinylpyrrolidone, tablet decomposition products (e.g., swellable crosslinked polymers, such as crosslinked carboxymethylcellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphoric acid or citrate buffer), and foaming agents, such as citrate / bicarbonate mixtures. Such excipients are well known and do not need to be discussed in detail here. 【0103】 Tablets can be designed to release the drug either by contact with gastric juice (immediate-release tablets) or by a controlled release over a long period of time or in a controlled manner within a specific region of the gastrointestinal tract (controlled-release tablets). 【0104】 Pharmaceutical compositions typically contain approximately 1% (w / w) to approximately 95% (w / w) of the active ingredient and 99% (w / w) to 5% (w / w) of pharmaceutically acceptable excipients (e.g., as defined above) or combinations of such excipients. Preferably, the composition contains approximately 20% (w / w) to approximately 90% (w / w) of the active ingredient and 80% (w / w) to 10% (w / w) of pharmaceutically acceptable excipients or combinations of excipients. Pharmaceutical compositions contain approximately 1% (w / w) to approximately 95% (w / w), preferably approximately 20% (w / w) to approximately 90% (w / w) of the active ingredient. Pharmaceutical compositions according to the present invention may be in unit dose form, for example, in the form of ampoules, vials, suppositories, pre-filled syringes, sugar-coated tablets, powders, tablets or capsules. 【0105】 Tablets and capsules may contain, for example, 0-20% (w / w) of tablet decomposition products, 0-5% (w / w) of lubricants, 0-5% (w / w) of flow aids, and / or fillers / or bulking agents (depending on the drug dose) of 0-99% (w / w). They may also contain 0-10% (w / w) of polymer binders, 0-5% (w / w) of antioxidants, and 0-5% (w / w) of pigments. Sustained-release tablets may further contain 0-99% (w / w) of controlled-release (e.g., delayed-release) polymers (depending on the dose). Film coatings of tablets or capsules typically contain 0-10% (w / w) of polymers, 0-3% (w / w) of pigments, and / or 0-2% (w / w) of plasticizers. 【0106】 The composition may be a parenteral composition. The parenteral formulation may contain 0-20% (w / w) of buffer, 0-50% (w / w) of cosolvent, and / or 0-99% (w / w) of water for injection (WFI) (depending on the dose and if lyophilized). The intramuscular depot formulation may also contain 0-99% (w / w) of oil. 【0107】 The composition may be in a form suitable for intranasal or intrabronchial administration. Such a composition should be suitable for atomization, which allows for inhalation through the mouth and facilitates absorption through the thin mucosa lining the nasal cavity. 【0108】 Many orally administered drugs may also be administered rectally as suppositories. The composition may be in a form suitable for rectal administration. In this form, the composition may contain a waxy substance that dissolves or liquefies after being inserted into the rectum. Such compositions may be prescribed to individuals who cannot take medication orally due to nausea, inability to swallow, or dietary restrictions, such as in many pre- and post-surgical cases. 【0109】 Pharmaceutical preparations may be provided to patients in a “patient pack” containing the entire course of treatment in a single package, usually a blister pack. 【0110】 Since the compound of formula (1) can usually be provided in unit dose form, it can typically contain enough of the compound to provide the desired level of physiological activity. For example, a formulation may contain 1 nanogram to 2 grams of the active ingredient, e.g., 1 nanogram to 2 milligrams of the active ingredient. Within these ranges, a particular subrange of the compound is 0.1 milligrams to 2 grams (more typically 10 milligrams to 1 gram, e.g., 50 milligrams to 500 milligrams) of the active ingredient, or 1 microgram to 20 milligrams (e.g., 1 microgram to 10 milligrams, e.g., 0.1 milligrams to 2 milligrams of the active ingredient). 【0111】 For oral compositions, the unit dose may contain 1 to 2 grams of the active compound, more typically 10 to 1 gram, for example 50 to 1 gram, or for example 100 to 1 gram. 【0112】 In some embodiments, the pharmaceutical composition comprises the free acid of compound A or the crystalline form of the tromethamine salt of compound A. 【0113】 In some embodiments, the pharmaceutical composition comprises tromethamine salt hydrate I of compound A. In some embodiments, the pharmaceutical composition comprises tromethamine salt hydrate II of compound A. In some embodiments, the pharmaceutical composition comprises tromethamine salt hydrate I of compound A and tromethamine salt hydrate I of compound A. 【0114】 The active compound may be administered to a patient in need (e.g., a human or animal patient) in an effective dose sufficient to achieve the desired therapeutic effect. The exact amount of the compound to be administered can be determined by the supervising physician following standard procedures. 【0115】 Method for preparing the compound of formula (1) The compounds of formula (1) can be prepared according to the synthetic methods described herein, some of which should be known to those skilled in the art. The present invention provides a method for preparing the compounds specified in formula (1) above. 【0116】 The compound of formula (1) can be prepared as described in Scheme 1 below. 【0117】 [ka] 【0118】 Thus, the substituted aminoaromatic nitrile of formula (2) is either commercially available or readily available from commercially available materials, and is usually converted to the tetrazole intermediate of formula (3) by heating in a solvent, such as DMF, DMSO, or 2-propanol, in the presence of sodium azide and ammonium chloride or zinc chloride, and at a temperature in the range of 110-130°C. Alternatively, the tetrazole of formula (3) can be obtained from the substituted 3-nitrobenzonitrile of formula (5), which is either commercially available or readily available from commercially available materials. The conversion of the nitrile functional group to provide the corresponding tetrazole of formula (6) is carried out as described above, and the subsequent reduction of the nitro group is usually brought about by zinc powder in the presence of 1,4-dioxane and ammonium chloride in water, while heating to reflux. The aniline group of the compound of formula (3) is then functionalized by a reaction using diethyl squalate in a solvent, such as EtOH or DCM, in the presence of a base, usually triethylamine. The ester functional group of squalate (4) is then hydrolyzed in a mixture of aqueous HCl and THF at a mild temperature, usually 60°C, to give the example of formula (1). Alternatively, the substituted aniline of formula (3) may be directly converted to the example of formula (1) by treatment with squalate (1), usually in refluxed water. 【0119】 Alternatively, the compound of formula (1) can be prepared as described in Scheme 2 below. The aryl bromide intermediate of formula (8) can be prepared from the aminoaniline of formula (7) as described in Scheme 1. Subsequent Suzuki reactions of the aryl bromide of formula (8) with a suitable boronic acid or boronic acid ester coupling partner (R=H or alkyl) yield examples of formula (1). The Suzuki reaction is usually carried out under microwave irradiation in a suitable solvent system, usually a combination of MeCN and H2O, in the presence of a base, e.g., K2CO3, and a catalytic palladium source, e.g., [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II). 【0120】 [ka] 【0121】 Scheme 3 is the compound of formula (1) (wherein R 2 Depending on the case, -CONHCH2R 4 -CONHCH2CH2OR 4 , -CONHR 4 Or -CON(CH3)R 4 This demonstrates the versatility of the preparation of the phenyl compounds (which are substituted with ). The biarylcarboxylic acid compounds of formula (9) are readily available from commercially available 3-amino-5-bromobenzonitrile (7) and are converted to the corresponding tetrazole compounds of formula (10) as described in Scheme 1. The aniline group of the intermediate compound of formula (10) is then functionalized as described in Scheme 1. The carboxylic acid group of the compound of formula (11) is then converted to an amide in the presence of a suitable amine, a coupling agent, e.g., 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxidehexafluorophosphate, a base, e.g., N,N-diisopropylethylamine, and a solvent, e.g., DMF. The ester functional group of cyclobuta-3-ene-1,2-dione of formula (12) is then hydrolyzed to yield an example of formula (13) (formula (1) (wherein R 2 Depending on the case, -CONHCH2R 4-CONHCH2CH2OR 4 , -CONHR 4 Or -CON(CH3)R 4 (corresponding to phenyl substituted with ). 【0122】 [ka] 【0123】 Here, X is N, and R 1 In the case where is H, the compound of formula (2b) can be prepared as described in scheme 4. Thus, commercially available 2,6-dichloroisonicotinonitrile (14) is heated to a temperature in the range of 110-120°C while (4-methoxyphenyl)methaneamine and S in a solvent, for example, NMP. N The Ar reaction is then carried out. The nitrile functional group of the intermediate of formula (15) is converted to a tetrazole group as shown in scheme 1. 2 If is an alkoxy group or a phenoxy group, the chlorine group of pyridine (16) is replaced with a suitable alcohol or metal alkoxide (e.g., NaOMe) in the presence or absence of a base, such as K2CO3, in a solvent, such as DMSO, and at temperatures ranging from room temperature to 120°C, to give the alkoxy or phenoxy-substituted compound (17). Alternatively, R 2If is phenyl, the Suzuki reaction of pyridyl chloride of formula (16) with a suitable phenylboronic acid or phenylborone ester yields intermediate (17). The Suzuki reaction is usually carried out in a solvent, e.g., 1,4-dioxane, at a temperature of 120°C, in the presence of a base, e.g., K2CO3, a palladium catalyst source, usually [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II). It will be understood by those skilled in the art that the sequence of steps developed in Scheme 4 can be completed in a different order than shown without affecting the overall success rate of the synthesis of the desired compound of formula (2b). The deprotection of the amino functional group of the compound of formula (17) is achieved by treatment with TFA in an acid, usually at 70°C. The conversion of the intermediate of formula (18) to give an example of formula (2b) is achieved as described in Scheme 1. 【0124】 [ka] 【0125】 In the specific reactions described above, it may be necessary to protect one or more groups to prevent the reaction from occurring at undesirable locations on the molecule. Examples of protecting groups and methods for protecting and deprotecting functional groups can be found in Greene's Protective Groups in Organic Synthesis, Fifth Edition, Editor: Peter GM Wuts, John Wiley, 2014, (ISBN:9781118057483). 【0126】 The compounds prepared by the aforementioned method may be isolated and purified by any of the various methods well known to those skilled in the art, including recrystallization and chromatographic techniques, such as column chromatography (e.g., flash chromatography), HPLC, and SFC. 【0127】 Normal procedure If a preparation route is not included, the relevant intermediates are commercially available. Commercial reagents were used without further purification. The final compounds and intermediates are named using ChemDraw Professional, Version 17.0.0.206(121). Room temperature (RT) refers to approximately 20–27°C. ¹H NMR spectra were recorded at 400 MHz or 500 MHz using either a Bruker, Varian, or Jeol instrument. Chemical shift values ​​are expressed in parts per million (ppm), i.e., (δ)-values ​​relative to the following solvents: chloroform - d=7.26 ppm, DMSO - d6=2.50 ppm, methanol - d4=3.31 ppm. The following abbreviations are used for the multiplicity of the NMR signal: s = singleline, br = broadline, d = doubleline, t = tripleline, q = quadrupleline, m = multiline. Coupling constants are listed as J values ​​and measured in Hz. The results of NMR and mass spectrometry were corrected for background peaks. Chromatography refers to column chromatography performed under nitrogen pressure (flash chromatography) conditions using 60–120 mesh or 40–633 μm, 60 Å silica gel. Microwave-mediated reactions were carried out in a Biotage Initiator or CEM Discover microwave reactor. 【0128】 LCMS analysis LC-MS analysis of the compounds was performed under electrospray conditions using the apparatus and method described below: 【0129】 LCMS method A Instrument: HP1100 with G1315A DAD, Micromass ZQ; Column: Phenomenex Gemini-NXC-18, 3 microns, 2.0 × 30 mm; Gradient [time (min) / B in solvent A (%)]: 0.00 / 2, 0.10 / 2, 2.50 / 95, 3.50 / 95; Solvent: Solvent A = 2.5 L H2O + 2.5 mL 28% ammonia H2O solution; Solvent B = 2.5 L MeCN + 135 mL H2O + 2.5 mL 28% ammonia H2O solution. Injection volume 1 μL; UV detection 230~400 nm; Mass detection 130~800 AMU; Column temperature 45 °C; Flow rate 1.5 mL / min. 【0130】 LCMS method B Instrument: Agilent 1260 Infinity LC with diode array detector, Agilent 6120B single quadrupole MS with API-ES Source, Column: Phenomenex Gemini-NXC-18, 3 micron, 2.0 × 30 mm, Gradient [time (min) / B in solvent A (%)]: 0.00 / 5, 2.00 / 95, 2.50 / 95, 2.60 / 5, 3.00 / 5, Solvent: Solvent A = 2.5 L H2O + 2.5 mL H2O solution with 28% NH3, Solvent B = 2.5 L MeCN + 129 mL H2O + 2.7 mL (H2O solution with 28% NH3), Injection volume 0.5 μL, UV detection 190~400 nm, Mass detection 130~800 AMU, Column temperature 40 °C, Flow rate 1.5 mL / min. 【0131】 LCMS Method C and LCMS Method D Apparatus: Agilent 1260 Infinity LC with diode array detector, Agilent 6120B single quadrupole MS with API-ES Source, Column: Restek, pentafluorophenylpropyl, 3 micron, 2.1 × 30 mm. Gradient [time (min) / B in solvent A (%)]: Method C: 0.00 / 5, 2.00 / 95, 2.50 / 95, 2.60 / 5, 3.00 / 5 or Method D: 0.00 / 2, 0.1 / 2, 8.4 / 95, 10 / 95, 10.1 / 2, 12 / 2, Solvent: Solvent A = Water (2.5 L) containing 2.5 mL formic acid, Solvent B = MeCN (2.5 L) containing 125 mL water and 2.5 mL formic acid. Injection volume 0.5 μL, UV detection 190-400 nm, mass detection 130-800 AMU, column temperature 40°C, flow rate 1.5 mL / min. 【0132】 LCMS method E Apparatus: Waters Acquity H Class, photodiode array, SQ detector; Column: BEH C18, 1.7 micron, 2.1 × 50 mm; Gradient [time (min) / B in solvent A (%)]: 0.00 / 5, 0.40 / 5, 0.8 / 35, 1.20 / 55, 2.50 / 100, 3.30 / 100, 4.00 / 5; Solvent: Solvent A = 5 mM ammonium acetate and 0.1% formic acid in H2O solution; Solvent B = 0.1% formic acid in MeCN solution; Injection volume: 2 μL; UV detection: 200-400 nm; Mass detection: 100-1200 AMU; Column at ambient temperature; Flow rate: 0.55 mL / min. 【0133】 LCMS method F Instrument: Agilent 1100 Series with DAD / ELSD, Agilent LC / MSDVL (G1956A), SL (G1956B) mass spectrometer; Column: Zorbax SB-C18, 1.8 micron, 4.6 × 15 mm; Gradient [Time (min) / B in Solvent A (%)]: 0.0 / 0, 1.5 / 100, 1.8 / 100, 1.81 / 0; Solvent: Solvent A = water and 0.1% formic acid, Solvent B = MeCN and 0.1% formic acid; Injection volume 1 μL; UV detection 200~400 nm; Mass detection 80~1000 AMU; Column at ambient temperature; Flow rate 3.0 mL / min. 【0134】 LCMS method G Instrument: Waters 2690 with PDA Detector 996, Acquity QDA mass-spectrometer; Column: X-BRIDGE C18, 5.0 micron, 4.6 × 100 mm; Gradient [time (min) / B in solvent A (%)]: 0.0 / 10, 1.0 / 10, 5.0 / 100, 7.0 / 100, 7.50 / 10, 8.0 / 10; Solvent: Solvent A = 0.1% formic acid and 10 mM ammonium bicarbonate aqueous solution; Solvent B = MeCN; Injection volume 1 μL; UV detection 190~800 nm; Mass detection 30~1250 AMU; Column temperature 35°C; Flow rate 1.2 mL / min. 【0135】 LCMS method H Apparatus: Waters Acquity H Class with photodiode array, SQ detector mass-spectrometer; Column: BEH C18, 1.7 micron, 2.1 × 50 mm; Gradient [time (min) / B in solvent A (%)]: 0.0 / 5, 0.4 / 5, 0.8 / 35, 1.2 / 55, 2.5 / 100, 3.3 / 100, 3.31 / 5, 4.0 / 5; Solvent: Solvent A = 0.1% formic acid and 5 mM ammonium acetate aqueous solution; Solvent B = 0.1% formic acid in MeCN solution; Injection volume 1 μL; UV detection 200~400 nm; Mass detection 100~1200 AMU; Column at ambient temperature; Flow rate 0.55 mL / min. 【0136】 LCMS Method I and Method J Apparatus for Method I: Shimadzu Nexera with photodiode array, LCMS-2020 mass-spectrometer or Apparatus for Method J: Agilent 1290 RRLC with photodiode array, Agilent 6120 mass-spectrometer, Column: X-BRIDGE C18, 3.5 microns, 4.6 × 50 mm, Gradient [Time (min) / B in Solvent A (%)]: 0.0 / 5, 5.0 / 90, 5.8 / 95, 7.20 / 95, 7.21 / 5, 10.0 / 5, Solvent: Solvent A = 0.1% aqueous ammonia solution, Solvent B = 0.1% ammonia MeCN solution, Injection volume 1 μL, UV detection 200~400 nm, Mass detection 60~1000 AMU, Column at ambient temperature, Flow rate 1.0 mL / min. 【0137】 LCMS method K Instrument: Waters 2690 with PDA Detector 996, Acquity QDA mass-spectrometer; Column: X-BRIDGE C18, 5.0 micron, 4.6 × 100 mm; Gradient [time (min) / B in solvent A (%)]: 0.0 / 10, 3.0 / 10, 6.0 / 100, 7.0 / 100, 7.01 / 10, 10.0 / 10; Solvent: Solvent A = 0.1% formic acid aqueous solution, Solvent B = MeOH; Injection volume: 1 μL; UV detection: 190~800 nm; Mass detection: 30~1250 AMU; Column temperature: 35°C; Flow rate: 1.0 mL / min. 【0138】 LCMS method L Instrument: Shimadzu LCMS-2010EV single quadrupole mass spectrometer with Shimadzu SPD-M20 APDA; Column: Atlantis C18, 3.0 micron, 4.6 × 50 mm; Gradient [time (min) / B in solvent A (%)]: 0.0 / 30, 3.0 / 90, 6.0 / 90, 6.1 / 30; Solvent: Solvent A = 0.1% formic acid aqueous solution, Solvent B = MeCN; Injection volume: 1 μL; UV detection: 254 nm; Mass detection: 80~800 AMU; Column temperature: 40°C; Flow rate: 0.8 mL / min. 【0139】 LCMS method M Instrument: Shimadzu LCMS-2010EV single quadrupole mass spectrometer with Shimadzu SPD-M20 APDA; Column: Capcell pack C18, 3.0 micron, 4.6 × 150 mm; Gradient [time (min) / B in solvent A (%)]: 0.0 / 30, 5.0 / 98, 9.5 / 98, 11.5 / 3, 12 / 3; Solvent: Solvent A = 0.1% formic acid aqueous solution, Solvent B = MeCN; Injection volume 1 μL; UV detection 254 nm; Mass detection 80~800 AMU; Column temperature 40°C; Flow rate 0.8 mL / min. 【0140】 abbreviation Bn = Benzyl CDI = Carbonyldiimidazole DCM = Dichloromethane DIPEA = N,N-diisopropylethylamine DMF = Dimethylformamide DMSO = Dimethyl sulfoxide ESI = Electrospray Ionization alkyl = ethyl acetate EtOH = Ethanol h=time HATU = (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxidehexafluorophosphate) H2O = Water HCl = hydrogen chloride, hydrochloric acid (prep)HPLC = (Preparative) High-Performance Liquid Chromatography IPA = Propane-2-ol K2CO3 = potassium carbonate LC = Liquid Chromatography MeCN = Acetonitrile MeOH = methanol Minutes = Minutes MS=mass spectrometry nm = nanometer NMP = N-methyl-2-pyrrolidone NMR=Nuclear Magnetic Resonance POCl3 = phosphate chloride RT=room temperature sat. = saturated SFC = Supercritical Fluid Chromatography TEA = Triethylamine TFA = Trifluoroacetic acid THF = Tetrahydrofuran TLC = Thin-Layer Calculation 【0141】 Standard synthesis procedure for intermediates Route 1 Preparation procedure for the intermediate 3,5-amino-[1,1'-biphenyl]-3-carbonitrile 【0142】 [ka] Step (i): A solution of intermediate 1,3-amino-5-bromobenzonitrile (200 mg, 1.02 mmol), intermediate 2, phenylboronic acid (161 mg, 1.32 mmol), K2CO3 (281 mg, 2.03 mmol), and Pd(Ph3P)4 (60 mg, 0.05 mmol) in 1,4-dioxane (4 mL) and water (1 mL) was heated to 100 °C for 3 hours under microwave irradiation and diluted with ethyl acetate and water. The organic compounds were separated, washed with brine, passed through hydrophobic frit, dried, and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of ethyl acetate (0%~45%) in PE to obtain intermediate 3,5-amino-[1,1'-biphenyl]-3-carbonitrile (165 mg, 0.85 mmol, yield 85%) as an orange solid. The data can be viewed in Table 2. 【0143】 Route 2 Preparation procedure for intermediate 5, ethyl 3'-amino-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-carboxylate 【0144】 [ka] Step (i): To a solution of intermediate 4, ethyl 3'-amino-5'-cyano-[1,1'-biphenyl]-3-carboxylate (73 mg, 0.27 mmol) in DMF (0.5 mL), NaN3 (36 mg, 0.55 mmol) and NH4Cl (30 mg, 0.55 mmol) were added. The mixture was heated at 130 °C for 3 hours, then cooled to RT, and the pH was acidified to 3-4 using 1 M HCl (aq). The precipitate was collected by filtration and dried under vacuum to obtain intermediate 5, ethyl 3'-amino-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-carboxylate (76 mg, 0.25 mmol, yield 91%) as a pink solid. The data can be viewed in Table 2. 【0145】 Route 3 Preparation procedure for intermediate 12, N-benzyl-3'-((2-ethoxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide 【0146】 [ka] Step (i): A mixture of intermediates 7,3-bromo-5-nitrobenzonitrile (1.0 g, 4.41 mmol), intermediate 6,4-(N-benzylaminocarbonyl)phenylboronic acid (1.24 g, 4.85 mmol), K2CO3 (1.22 g, 8.81 mmol), and Pd(Ph3P)4 (0.15 g, 0.130 mmol) in 1,4-dioxane (17 mL) and water (2 mL) was heated to 100°C for 1 hour under microwave irradiation. After this time, the mixture was diluted with SiO2, washed with water and then brine, and concentrated by passing through hydrophobic frit and drying. The crude product was triturated with Et2O, and the resulting solid was recovered by filtration to obtain intermediate 8, N-benzyl-3'-cyano-5'-nitro-[1,1'-biphenyl]-4-carboxamide (1.12 g, 3.13 mmol, yield 71%), as a tan-colored solid. The data can be viewed in Table 2. 【0147】 Step (ii): A mixture of intermediate 8, N-benzyl-4-(3-cyano-5-nitrophenyl)benzamide (1.03 g, 2.87 mmol), NaN3 (373 mg, 5.74 mmol), and NH4Cl (307 mg, 5.74 mmol) in DMF (22 mL) was heated to 130 °C for 2 hours. The mixture was cooled to RT and the pH was acidified to 2 using 1 M HCl (aq). The solid was recovered by filtration, suspended in toluene, and concentrated (×2) to obtain intermediate 9, N-benzyl-4-[3-nitro-5-(1H-tetrazole-5-yl)phenyl]benzamide (950 mg, 2.37 mmol, yield 83%) as a tan-colored solid. The data can be viewed in Table 2. 【0148】 Step (iii): Intermediate 9, a mixture of 1,4-dioxane (44 mL) and water (6.3 mL) containing N-benzyl-4-[3-nitro-5-(1H-tetrazole-5-yl)phenyl]benzamide (630 mg, 1.57 mmol), zinc powder (412 mg, 6.29 mmol), and NH4Cl (842 mg, 15.7 mmol) was heated and refluxed for 4 hours. The mixture was cooled to RT and filtered through a Celite bed (the entire mixture was washed with MeOH). The filtrate was concentrated and the residue was dissolved in 1 M HCl (aq) (20 mL). The pH was adjusted to approximately 6 by adding 1 M NaOH (aq). The precipitate was recovered by filtration, washed with water, and then suspended and concentrated in toluene to obtain the intermediate 10,4-[3-amino-5-(1H-tetrazole-5-yl)phenyl]-N-benzyl-benzamide (575 mg, 1.55 mmol, yield 99%) as a tan-colored solid. The data can be viewed in Table 2. 【0149】 Step (iv): Intermediate 10, 4-[3-amino-5-(1H-tetrazole-5-yl)phenyl]-N-benzyl-benzamide (575 mg, 1.55 mmol), intermediate 11, diethyl squalate (423 mg, 2.48 mmol), and TEA (0.44 mL, 3.1 mmol) were mixed in an EtOH suspension and stirred at RT for 24 hours, then 1 M HCl (aq) was added. The precipitate was collected by filtration, suspended in toluene, and concentrated to obtain intermediate 12, N-benzyl-3'-((2-ethoxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (635 mg, 1.28 mmol, yield 83%) as a tan solid. The data can be viewed in Table 2. 【0150】 Route 4 Preparation procedure for intermediate 15, 3'-amino-5'-cyano-N-(2-methoxyethyl)-[1,1'-biphenyl]-4-carboxamide 【0151】 [ka] Step (i): Intermediate 13, 3'-amino-5'-cyano-[1,1'-biphenyl]-4-carboxylic acid (400 mg, 1.68 mmol), intermediate 14, 2-methoxyethane-1-amine (0.3 mL, 3.36 mmol), and DIPEA (0.88 mL, 5.04 mmol) were dissolved in DMF (4.8 mL), to which HATU (768 mg, 2.02 mmol) was added. The mixture was stirred at RT for 1 hour, then diluted with SiO2, washed with water and brine, and concentrated by passing through hydrophobic frit and drying. The crude product was purified by flash column chromatography (reverse phase, C18) under a gradient of MeCN (10%~50%) in water (containing 0.1% v / v HCOOH) to obtain intermediate 15, 3'-amino-5'-cyano-N-(2-methoxyethyl)-[1,1'-biphenyl]-4-carboxamide (317 mg, 1.07 mmol, yield 64%) as a tan solid. The data can be viewed in Table 2. 【0152】 Route 5 Preparation procedure for intermediate 17, 4-chloro-3-(1H-tetrazole-5-yl)aniline 【0153】 [ka] Step (i): A solution of intermediate 16, 5-amino-2-chlorobenzonitrile (400 mg, 2.63 mmol), NaN3 (342 mg, 5.26 mmol), and NH4Cl (281 mg, 5.88 mmol) in DMF (3 mL) was heated to 130 °C for 3 days. The mixture was cooled, insoluble material was removed by filtration, and the mixture was washed with MeCN. The solution was concentrated under reduced pressure, the residue was dissolved in siRNA, and the mixture was washed with water. The organic layer was dried, and the solvent was evaporated. The residue was crystallized from DCM to obtain intermediate 17, 4-chloro-3-(1H-tetrazole-5-yl)aniline. The data can be viewed in Table 2. 【0154】 Route 6 Procedure for preparing intermediate 21,3-cyclopropyl-5-(1H-tetrazole-5-yl)aniline 【0155】 [ka] Step (i): Intermediate 7, 3-bromo-5-nitrobenzonitrile (3.00 g, 13.3 mmol), intermediate 18, (2-cyclopropyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane) (3.34 g, 19.9 mmol), and K2CO3 (3.68 g, 26.62 mmol) were suspended in 1,4-dioxane (22 mL) and water (8 mL) at room temperature and degassed with N2 for 15 minutes. Then, PdCl2 (dppf) (0.97 g, 1.32 mmol) was added and the mixture was heated to 100 °C for 4 hours. The mixture was cooled and partitioned between water (300 mL) and butyl (200 mL). The organic layer was separated and the aqueous layer was further extracted with butyl (2 × 100 mL). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of siRNA (0%-5%) in PE to obtain intermediate 19,3-cyclopropyl-5-nitrobenzonitrile (1.30 g, 6.91 mmol, yield 52%) as a yellow solid. The data can be viewed in Table 2. 【0156】 Step (ii): Intermediate 19, 3-cyclopropyl-5-nitrobenzonitrile (1.30 g, 6.91 mmol) was dissolved in MeOH (7 mL), and a suspension of NH4Cl (2.01 g, 37.2 mmol) and zinc powder (2.43 g, 37.22 mmol) in water (7 mL) was added. The reaction was stirred at RT for 30 minutes, and then filtered through Celite. The filtrate was concentrated, and the residue was partitioned between sat. aq. NaHCO3 and IgG. The organic layer was separated, and the aqueous layer was further extracted with IgG (×2). The combined organic layer was dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of IgG in PE (0%~30%) to obtain intermediate 20, 3-amino-5-cyclopropylbenzonitrile (1.01 g, 6.39 mmol, yield 93%) as a yellow liquid. The data can be viewed in Table 2. 【0157】 Step (iii): Intermediate 20, a suspension of 2-propanol containing 3-amino-5-cyclopropylbenzonitrile (1.01 g, 6.39 mmol), NaN3 (2.07 g, 31.9 mmol), and ZnCl2 (4.35 g, 31.9 mmol) was heated to 130°C for 16 hours. The mixture was cooled, filtered through Celite, and the filtrate was concentrated. The residue was partitioned between water and ethyl acetate. The organic layer was separated, and the aqueous layer was further extracted with ethyl acetate (×2). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (reverse phase, C18) under a gradient of 0.1N HCOOHaq. (0%~30%) in MeCN to obtain intermediate 21, 3-cyclopropyl-5-(1H-tetrazole-5-yl)aniline (800 mg, 3.98 mmol, yield 62%). The data can be viewed in Table 2. 【0158】 Route 7 Procedure for preparing intermediate 23,3-amino-5-cyclohexylbenzonitrile 【0159】 [ka] Step (i): Intermediate 22, a solution of 5-nitro-2',3',4',5'-tetrahydro-[1,1'-biphenyl]-3-carbonitrile (3.00 g, 13.2 mmol) and 10% Pd / C, 50% water (3.00 g) in MeOH (30 mL) was stirred at RT under an H2 gas atmosphere for 5 hours. After this time, the mixture was filtered through Celite and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of Âxy (0%~25%) in DCM to obtain intermediate 23, 3-amino-5-cyclohexylbenzonitrile (1.20 g, 6.0 mmol, yield 45%) as a yellow liquid. The data can be viewed in Table 2. 【0160】 Route 8 Procedure for preparing intermediate 25,3-nitro-5-(pyridazin-4-yl)benzonitrile 【0161】 [Chemical formula] Step (i): A MeCN suspension of intermediate 7, 3-bromo-5-nitrobenzonitrile (0.35 g, 1.55 mmol), intermediate 24, 4-(tributylstannyl)pyridazine (0.57 g, 1.55 mmol) and CsF (0.47 g, 3.10 mmol) was degassed with N2 for 15 minutes, then Pd(PPh3)4 (0.09 g, 0.80 mmol) and copper(I) iodide (0.06 g, 0.31 mmol) were added. The mixture was heated to 40 °C for 4 hours, then cooled and partitioned between EtOAc and water. The organic compound was separated and the aqueous layer was extracted with EtOAc. The combined organic compounds were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of EtOAc in hexane (0% - 40%) to give intermediate 25, 3-nitro-5-(pyridazin-4-yl)benzonitrile (0.25 g, 1.10 mmol, 71%) as a yellow solid. The data can be viewed in Table 2. 【0162】 Route 9 Procedure for the preparation of intermediate 29, 3'-((2-ethoxy-3,4-dioxocyclobut-1-en-1-yl)amino)-5'-(1H-tetrazol-5-yl)-[1,1'-biphenyl]-4-carboxylic acid 【0163】 [Chemical formula] Step (i): A solution of intermediate 1,3-amino-5-bromobenzonitrile (20.0 g, 101.5 mmol), intermediate 26, (4-(ethoxycarbonyl)phenyl)boronic acid (23.6 g, 121.8 mmol), K2CO3 (28.0 g, 203 mmol), and Pd(Ph3P)4 (2.34 g, 2.03 mmol) in 1,4-dioxane (200 mL) and water (30 mL) was heated to 90 °C for 16 hours, then cooled and partitioned between SiO2 and water. The organic compounds were separated, washed with brine, dried over Na2SO4, and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of siRNA (0%-16%) in hexane to obtain intermediate 27, ethyl 3'-amino-5'-cyano-[1,1'-biphenyl]-4-carboxylate (23.0 g, 86.5 mmol, yield 85%), as an off-white solid. The data can be viewed in Table 2. 【0164】 Step (ii): Intermediate 27, ethyl 3'-amino-5'-cyano-[1,1'-biphenyl]-4-carboxylate (23.0 g, 86.5 mmol) was dissolved in THF (130 mL) and water (130 mL), to which lithium hydroxide monohydrate (6.22 g, 259.3 mmol) was added. The mixture was stirred at RT for 6 hours, then concentrated to dryness. 1N HCl (aq) was added, and the resulting precipitate was collected by filtration. The solid was dried under vacuum to obtain intermediate 13, 3'-amino-5'-cyano-[1,1'-biphenyl]-4-carboxylic acid (17.0 g, 71.4 mmol, yield 83%) as an off-white solid. The data can be viewed in Table 2. 【0165】 Step (iii): A solution of intermediate 13, 3'-amino-5'-cyano-[1,1'-biphenyl]-4-carboxylic acid (16.0 g, 67.2 mmol), NaN3 (8.74 g, 134.4 mmol), and NH4Cl (7.19 g, 134.4 mmol) in DMF (80 mL) was heated to 130 °C for 16 hours, then cooled, and 1N HCl (aq) was added to acidify the pH to 3. The resulting precipitate was collected by filtration and dried under vacuum to obtain intermediate 28, 3'-amino-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxylic acid (13.0 g, 46.3 mmol, yield 69%) as an off-white solid. The data can be viewed in Table 2. 【0166】 Step (iv): To a solution of intermediate 28, 3'-amino-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxylic acid (12.5 g, 44.4 mmol) in EtOH (120 mL), TEA (12.4 mL, 88.9 mmol) and intermediate 11, diethyl squalate (7.50 g, 44.4 mmol) were added. The mixture was stirred at RT for 16 hours, and then acidified with 1N HCl(aq). The resulting precipitate was collected by filtration and dried under vacuum to obtain intermediate 29, 3'-((2-ethoxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxylic acid (15.0 g, 37.0 mmol, yield 83%) as a brown solid. The data can be viewed in Table 2. 【0167】 Route 10 Preparation procedure for intermediate 31, 3-((3-bromo-5-(1H-tetrazole-5-yl)phenyl)amino)-4-ethoxycyclobuta-3-en-1,2-dione 【0168】 [ka] Step (i): Intermediate 1, 3-amino-5-bromobenzonitrile (26.0 g, 132 mmol) was suspended in 2-propanol (500 mL) and NaN3 (43.1 g, 663.3 mmol) and ZnCl2 (90.4 g, 663.3 mmol) were added. The mixture was stirred at 130 °C for 4 hours and then filtered through Celite. The filtrate was concentrated and the residue was partitioned between RINKAN and sat. aq. NaHCO3. The organic layer was separated, and the aqueous layer was further extracted with RINKAN (×3). The combined organic compounds were dried over Na2SO4 and concentrated to obtain intermediate 30, 3-bromo-5-(1H-tetrazole-5-yl)aniline (26.8 g, 112.2 mmol, yield 85%) as a yellow solid, which was used without further purification. The data can be viewed in Table 2. 【0169】 Step (ii): To a solution of intermediate 30, 3-bromo-5-(1H-tetrazole-5-yl)aniline (10.0 g, 27.5 mmol) in DCM (100 mL), TEA (11.7 mL, 83.6 mmol) was added. After stirring the mixture for 10 minutes, intermediate 11, diethyl squalate (6.18 mL, 41.8 mmol) was added. The mixture was stirred at RT for 4 hours and then concentrated under reduced pressure. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of MeOH (0%~15%) in DCM to obtain intermediate 31, 3-((3-bromo-5-(1H-tetrazole-5-yl)phenyl)amino)-4-ethoxycyclobuta-3-en-1,2-dione (3.80 g, 10.4 mmol, yield 38%) as an off-white solid. The data can be viewed in Table 2. 【0170】 Route 11 Preparation procedure for intermediate 35, (5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-((4-(trifluoromethyl)benzyl)oxy)pyridine) 【0171】 [ka] Step (i): To a solution of intermediate 33, [4-(trifluoromethyl)phenyl]methanol (3.00 g, 17.0 mmol) in DMF (30 mL), K2CO3 (3.53 g, 20.2 mmol) was added at RT. The mixture was stirred for 10 minutes, after which intermediate 32, 5-bromo-2-fluoropyridine (2.99 g, 17.0 mmol) was added. The mixture was heated to 80°C for 2 hours, then cooled and partitioned between butyl and water. The aqueous layer was further extracted with butyl, and the combined organic compounds were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of butyl (0%~5%) in hexane to obtain intermediate 34, (5-bromo-2-((4-(trifluoromethyl)benzyl)oxy)pyridine) (2.00 g, 6.02 mmol, yield 35%) as an off-white solid. The data can be viewed in Table 2. 【0172】 Step (ii): Intermediate 34, a solution of (5-bromo-2-((4-(trifluoromethyl)benzyl)oxy)pyridine (1.20 g, 3.61 mmol), KOAc (1.06 g, 10.9 mmol), bis(pinacolate)diborone (2.76 g, 10.9 mmol), and PdCl2 (dppf) (0.13 g, 0.18 mmol) in 1,4-dioxane (20 mL) was heated to 80°C under nitrogen for 4 hours, then cooled and partitioned between butyl and water. The aqueous layer was further extracted using butyl. The combined organic compounds were dried with Na2SO4 and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of ethyl phosphate (0%~8%) in hexane to obtain intermediate 35, (5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-((4-(trifluoromethyl)benzyl)oxy)pyridine) (1.20 g, 3.17 mmol, 87%) as a white semi-solid. The data can be viewed in Table 2. 【0173】 Route 12 Preparation procedure for intermediate 40, 2-benzyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole 【0174】 [ka] Step (i): A mixture of intermediate 36, 5-bromo-2-nitrobenzaldehyde (3.00 g, 13.1 mmol), and intermediate 37, benzylamine (1.82 g, 17.0 mmol), in toluene (30 mL) was heated to 80°C for 4 hours. The reaction was cooled to room temperature and concentrated to obtain crude intermediate 38, N-benzyl-1-(5-bromo-2-nitrophenyl)methanymine (2.10 g), as a dark brown liquid. The data can be viewed in Table 2. 【0175】 Step (ii): Intermediate 38, N-benzyl-1-(5-bromo-2-nitrophenyl)methanymine (2.10 g, 6.60 mmol) was dissolved in triethyl phosphite (3.40 mL, 19.8 mmol), and the mixture was heated to 210°C for 5 minutes under microwave irradiation. The mixture was partitioned between ethyl acetate and water and separated. The aqueous layer was extracted using ethyl acetate (×2), and the combined organic compounds were dried over Na2SO4 and then concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of ethyl acetate (0%~20%) in hexane to obtain intermediate 39, 2-benzyl-5-bromo-2H-indazole (1.30 g, 4.54 mmol, 35% yield after 2 steps) as a brown gum. The data can be viewed in Table 2. 【0176】 Step (iii): Intermediate 39, 2-benzyl-5-bromo-2H-indazole (1.30 g, 4.54 mmol), KOAc (1.34 g, 13.6 mmol), bis(pinacolato)diboron (3.46 g, 13.6 mmol), and PdCl2(dppf) (0.17 g, 0.22 mmol) were dissolved in 1,4-dioxane (15 mL) under a nitrogen atmosphere. The reaction mixture was stirred at 80 °C for 4 hours, then cooled to room temperature and partitioned between EtOAc and water. The organic compound was separated, and the aqueous layer was further extracted with EtOAc. The combined organic compounds were dried over Na2SO4 and concentrated to give Intermediate 40, 2-benzyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (1.0 g, 2.99 mmol, 66% yield) as a brown gum, which was used without further purification. The data can be viewed in Table 2. 【0177】 Route 13 Procedure for the preparation of Intermediate 44, 2-(4-(trifluoromethyl)phenoxy)ethan-1-amine, hydrochloride 【0178】 [Chemical formula] Step (i): To a solution of Intermediate 42, tert-butyl (2-bromoethyl)carbamate (876 mg, 3.9 mmol) in DMF (10 mL) were added Intermediate 41, 4-(trifluoromethyl)phenol (422 mg, 2.6 mmol) and Cs2CO3 (1.25 g, 3.8 mmol). The mixture was heated to 90 °C for 4 hours, then cooled and poured into water. The mixture was extracted with EtOAc, the organic compound was washed with brine, passed through a hydrophobic frit to dry, and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of EtOAc in petroleum ether (0% - 50%) to give Intermediate 43, tert-butyl (2-(4-(trifluoromethyl)phenoxy)ethyl)carbamate (793 mg, 2.6 mmol, 100% yield) as a yellow oil. The data can be viewed in Table 2. 【0179】 Step (ii): Intermediate 43, tert-butyl (2-(trifluoromethyl)phenoxy)ethyl) carbamate (793 mg, 2.6 mmol) in a solution of 1,4-dioxane (2 mL), was mixed with a solution of 1,4-dioxane (2 mL) in 4 M HCl. The mixture was stirred for 18 hours, after which MeOH (1 mL) was added. The mixture was stirred for a further 30 minutes, and then concentrated to obtain intermediate 44, 2-(4-(trifluoromethyl)phenoxy)ethane-1-amine hydrochloride (596 mg, 2.5 mmol, yield 95%), as a pale yellow solid. The data can be viewed in Table 2. 【0180】 Route 14 Preparation procedure for intermediate 48, 2-(2-(3,5-bis(trifluoromethyl)phenoxy)ethoxy)-5-bromopyridine 【0181】 [ka] Step (i): To a solution of intermediate 45, 3,5-bis(trifluoromethyl)phenol (5.00 g, 21.7 mmol) in DMF (50 mL), K2CO3 (8.99 g, 65.2 mmol) was added. The mixture was stirred at RT for 20 minutes, and then intermediate 46, 2-bromoethane-1-ol (9.25 mL, 130 mmol) was added. The mixture was heated to 80°C for 4 hours, then cooled and partitioned between ethyl acetate and water. The aqueous layer was further extracted with ethyl acetate, and the combined organic compounds were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of ethyl acetate (0%~20%) in hexane to obtain intermediate 47, 2-(3,5-bis(trifluoromethyl)phenoxy)ethane-1-ol (3.90 g, 14.2 mmol, yield 66%) as an oil. The data can be viewed in Table 2. 【0182】 Step (ii): To a suspension of NaH (60% in mineral oil) (0.77 g, 19.2 mmol) in DMF (25 mL) at 0°C, intermediate 47, 2-(3,5-bis(trifluoromethyl)phenoxy)ethane-1-ol (3.00 g, 11.0 mmol) was added. The mixture was heated in RT for 15 minutes, and then intermediate 32, 5-bromo-2-fluoropyridine (2.30 g, 13.1 mmol) was added. The reaction was stirred in RT for 1 hour, and then partitioned between butyl and water. The aqueous layer was further extracted with butyl, and the combined organic compounds were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of ethylethanol (0%-12%) in hexane to obtain intermediate 48, 2-(2-(3,5-bis(trifluoromethyl)phenoxy)ethoxy)-5-bromopyridine (3.60 g, 8.37 mmol, yield 77%) as a yellow semi-solid. The data can be viewed in Table 2. 【0183】 Route 15 Procedure for preparing intermediate 51, 2-(4-(trifluoromethyl)phenoxy)ethane-1-ol 【0184】 [ka] Step (i): Intermediate 41, 4-(trifluoromethyl)phenol (2.80 g, 17.3 mmol), intermediate 49, 2-((tert-butyldimethylsilyl)oxy)ethane-1-ol (4.00 g, 22.5 mmol), and triphenylphosphine (5.89 g, 22.5 mmol) were dissolved in toluene (30 mL) at 0°C, to which DIAD (4.40 mL, 22.5 mmol) was added dropwise. The mixture was stirred at RT for 16 hours, and then partitioned between butyl and water. The aqueous layer was further extracted with butyl, and the combined organic compounds were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of SiO (0%~2%) in hexane to obtain intermediate 50, tert-butyldimethyl(2-(4-(trifluoromethyl)phenoxy)ethoxy)silane (3.40 g, 10.6 mmol, yield 61%), as a yellow liquid. The data can be viewed in Table 2. 【0185】 Step (ii): To a solution of intermediate 50, tert-butyldimethyl(2-(4-(trifluoromethyl)phenoxy)ethoxy)silane (3.40 g, 10.6 mmol) in THF (30 mL) at 0°C, 1N HCl (aq) (30 mL) was added. The mixture was stirred at RT for 1 hour, and then partitioned between sat. aq. NaHCO3 and HCl. The organic compounds were separated, dried over Na2SO4, and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of HCl (0%~30%) in hexane to obtain intermediate 51, 2-(4-(trifluoromethyl)phenoxy)ethane-1-ol (2.50 g, 12.1 mmol, assumed quantitative) as a yellow liquid. The data can be viewed in Table 2. 【0186】 Route 16 Procedure for preparing intermediate 54, (4-((4-carbamoylbenzyl)carbamoyl)phenyl)boronic acid 【0187】 [ka] Step (i): Intermediate 52, 4-Boronobenzoic acid (795 mg, 4.79 mmol) was dissolved in ice-cold DMF (5 mL) and HATU (2.27 g, 5.99 mmol) was added. The mixture was stirred at the same temperature for 10 minutes, after which intermediate 53, 4-(aminomethyl)benzamide (600 mg, 3.99 mmol) and DIPEA (2.02 mL, 11.98 mmol) were added sequentially. The mixture was heated to RT and stirred for 6 hours, then partitioned between siRNA and water. The organic compounds were separated and the aqueous layer was washed with siRNA. The combined organic compounds were dried over Na2SO4 and concentrated to obtain intermediate 54, (4-((4-carbamoylbenzyl)carbamoyl)phenyl)boronic acid (0.350 g, 1.17 mmol, yield 29%) as a yellow solid. The data can be viewed in Table 2. 【0188】 Route 17 Preparation procedure for intermediate 60, N-(4-methoxybenzyl)-6-phenoxy-4-(1H-tetrazole-5-yl)pyridine-2-amine 【0189】 [ka] Step (i): To a solution of intermediate 56, phenol (1.31 g, 14.0 mmol) in DMSO (40 mL), K2CO3 (2.4 g, 17.4 mmol) was added. The mixture was stirred at RT for 30 minutes, then intermediate 55, 2,6-dichloroisonicotinonitrile (2.0 g, 11.6 mmol) was added. The mixture was stirred at RT for 24 hours, then poured into water and extracted with RINKAN (×3). The combined organic compounds were washed with brine, dried over Na2SO4, and concentrated. The crude product was purified by flash column chromatography (normal phase, silica) under a gradient of RINKAN (0%~20%) in petroleum ether to obtain intermediate 57, 2-chloro-6-phenoxyisonicotinonitrile (2.5 g, 10.9 mmol, yield 94%) as a yellow liquid. The data can be viewed in Table 2. 【0190】 Step (ii): A solution of intermediate 57, 2-chloro-6-phenoxyisonicotinonitrile (2.5 g, 10.8 mmol) and intermediate 58, 4-methoxybenzylamine (1.63 g, 11.9 mmol) in NMP (26 mL) was heated to 120 °C for 3 hours. The mixture was then cooled and poured into water. The precipitate was filtered, and the crude product was purified by flash column chromatography (normal phase, silica) under a gradient of toluene (10-50%) in petroleum ether to obtain intermediate 59, 2-((4-methoxybenzyl)amino)-6-phenoxyisonicotinonitrile (1.7 g, 5.1 mmol, yield 47%) as an off-white solid. The data can be viewed in Table 2. 【0191】 Step (iii): Intermediate 59, a solution of 2-((4-methoxybenzyl)amino)-6-phenoxyisonicotinonitrile (1.7 g, 5.13 mmol), NaN3 (1.33 g, 20.5 mmol), and NH4Cl (1.1 g, 20.5 mmol) in DMF (9.2 mL) was heated to 130 °C for 2 hours. The mixture was cooled and poured into water. The pH of the aqueous mixture was acidified to 1 with 1N HCl(aq), stirred for 30 minutes, and then extracted with ELISA (×3). The combined organic compounds were washed with brine, dried over Na2SO4, and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of MeOH (0%~5%) in DCM to obtain intermediate 60, N-(4-methoxybenzyl)-6-phenoxy-4-(1H-tetrazole-5-yl)pyridine-2-amine (1.6 g, 4.27 mmol, yield 83%) as a brown gum. The data can be viewed in Table 2. 【0192】 Route 18 Preparation procedure for intermediate 61,6-phenoxy-4-(1H-tetrazole-5-yl)pyridine-2-amine 【0193】 [ka] Step (iv): A solution of intermediate 60, N-(4-methoxybenzyl)-6-phenoxy-4-(1H-tetrazole-5-yl)pyridine-2-amine (1.6 g, 4.27 mmol) in TFA (16 mL) was heated to 70°C for 24 hours in a sealed tube, after which the mixture was cooled and concentrated to dryness. Intermediate 61, 6-methoxy-4-(1H-tetrazole-5-yl)pyridine-2-amine (0.75 g, 3.91 mmol, yield 93%) was obtained as a brown liquid by trituration of the crude product using DCM. The data can be viewed in Table 2. 【0194】 Route 19 Preparation procedure for intermediate 63, 6-chloro-N-(4-methoxybenzyl)-4-(1H-tetrazole-5-yl)pyridine-2-amine 【0195】 [ka] Step (i): A solution of intermediate 55, 2,6-dichloroisonicotinonitrile (4.5 g, 26.2 mmol), and intermediate 58, 4-methoxybenzylamine (3.6 g, 26.2 mmol), in NMP (45 mL) was heated to 110 °C for 3 hours. The mixture was cooled and poured into ice water (500 mL). The resulting precipitate was filtered, washed with water, and dried under vacuum to obtain intermediate 62, 2-chloro-6-((4-methoxybenzyl)amino)isonicotinonitrile (3.6 g, 13.2 mmol, yield 50%) as an off-white solid. The data can be viewed in Table 2. 【0196】 Step (ii): A solution of intermediate 62, 2-chloro-6-((4-methoxybenzyl)amino)isonicotinonitrile (1.36 g, 5 mmol), NaN3 (650 mg, 10 mmol), and NH4Cl (535 mg, 10 mmol) in DMF (9 mL) was heated to 130 °C for 2 hours. The mixture was cooled and poured into water. The pH of the aqueous mixture was acidified to 1 with 1N HCl (aq), the resulting precipitate was filtered, washed with water, and dried under vacuum to obtain intermediate 63, 6-chloro-N-(4-methoxybenzyl)-4-(1H-tetrazole-5-yl)pyridine-2-amine (960 mg, 3.04 mmol, yield 61%) as an off-white solid. The data can be viewed in Table 2. 【0197】 Route 20 Preparation procedure for intermediate 64,6-methoxy-N-(4-methoxybenzyl)-4-(1H-tetrazole-5-yl)pyridine-2-amine 【0198】 [ka] Step (i): To a solution of intermediate 63, 6-chloro-N-(4-methoxybenzyl)-4-(1H-tetrazole-5-yl)pyridine-2-amine (1.0 g, 3.16 mmol) in DMSO (6.3 mL), NaOMe (30 wt% MeOH, 1.14 mL, 6.33 mmol) was added. The mixture was heated to 120 °C for 20 hours, then cooled and poured into water. The pH of the aqueous mixture was acidified to 1 using 1N HCl (aq), and the resulting precipitate was collected. The crude product was purified by flash column chromatography (normal phase, silica) under a chloroform-MeOH (0%~10%) gradient to obtain intermediate 64, 6-methoxy-N-(4-methoxybenzyl)-4-(1H-tetrazole-5-yl)pyridine-2-amine (700 mg, 2.24 mmol, yield 71%) as an off-white solid. The data can be viewed in Table 2. 【0199】 Route 21 Procedure for preparing intermediate 65, N-(4-methoxybenzyl)-6-phenyl-4-(1H-tetrazole-5-yl)pyridine-2-amine 【0200】 [ka] Step (i): A solution of intermediate 63, 6-chloro-N-(4-methoxybenzyl)-4-(1H-tetrazole-5-yl)pyridine-2-amine (1 g, 3.15 mmol), intermediate 2, phenylboronic acid (1.15 g, 9.46 mmol), K2CO3 (1.3 g, 9.46 mmol), and Pd(dppf)Cl2.DCM (257 mg, 0.32 mmol) in 1,4-dioxane (10 mL) was heated to 120 °C for 6 hours, after which the mixture was cooled and poured into water. The pH of the aqueous mixture was acidified to 1 using 1N HCl(aq), and the resulting precipitate was collected. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of MeOH (0%~8%) in chloroform to obtain intermediate 65, N-(4-methoxybenzyl)-6-phenyl-4-(1H-tetrazole-5-yl)pyridine-2-amine (810 mg, 2.26 mmol, yield 72%) as an off-white solid. The data can be viewed in Table 2. 【0201】 Route 22 Preparation procedure for intermediate 71, 3-ethoxy-4-[3-ethyl-5-(1H-tetrazole-5-yl)anilino]cyclobuta-3-en-1,2-dione 【0202】 [ka] Step (i): Intermediate 1,3-amino-5-bromobenzonitrile (1.0 g, 5.08 mmol), intermediate 6,6,4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (1.29 mL, 7.61 mmol), K2CO3 (1.40 g, 10.2 mmol), and Pd(Ph3P)4 (0.118 g, 0.10 mmol) were heated in 1,4-dioxane (24 mL) and water (6 mL) for 18 hours to 80°C, then cooled to RT and partitioned between SiO2 and water. The organic compounds were separated, washed with brine, passed through hydrophobic frit, dried, and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of siRNA (0%-25%) in isohexane to obtain the intermediate 67,3-amino-5-vinylbenzonitrile (695 mg, 4.82 mmol, 95% yield) as an orange solid. The data can be viewed in Table 2. 【0203】 Step (ii): Intermediate 68, benzyl chloroformate (0.95 mL, 6.7 mmol), was added to a solution of intermediate 67, 3-amino-5-vinylbenzonitrile (640 mg, 4.44 mmol), and K2CO3 (927 mg, 6.7 mmol) in THF (37 mL). The mixture was stirred at RT for 18 hours, and then partitioned between RINKAN and water. The organic compounds were separated, washed with brine, and dried by passing through hydrophobic frit to concentrate. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of Et2O (0%~60%) in isohexane to obtain intermediate 69, benzyl (3-cyano-5-vinylphenyl) carbamate (944 mg, 3.40 mmol, yield 76%), as an off-white solid. The data can be viewed in Table 2. 【0204】 Step (iii): A solution of intermediate 69, benzyl(3-cyano-5-vinylphenyl) carbamate (614 mg, 2.2 mmol), NaN3 (287 mg, 4.4 mmol), and NH4Cl (235 mg, 4.4 mmol) in DMF (6 mL) was heated to 130 °C for 2 hours. After this time, the mixture was cooled and partitioned between 1 M HCl (aq) and RINKAN. The organic compounds were separated, washed with brine, and dried by passing through hydrophobic frit to concentrate. The crude product was purified by flash column chromatography (reverse phase, C18) under a gradient of MeCN (10%~60%) in water (0.1% v / v HCOOH) to obtain intermediate 70, benzyl(3-(1H-tetrazole-5-yl)-5-vinylphenyl) carbamate (355 mg, 1.12 mmol, yield 50%) as a beige solid. The data can be viewed in Table 2. 【0205】 Step (iv): A mixture of intermediate 70, benzyl (3-(1H-tetrazole-5-yl)-5-vinylphenyl) carbamate (270 mg, 0.84 mmol), TEA (0.12 mL, 0.84 mmol), and 10% Pd / C (60 mg) in MeOH (4 mL) was stirred under a hydrogen gas atmosphere for 3.5 hours. The mixture was filtered through Celite (washed with SiO2) and concentrated. The residue was dissolved in EtOH (4 mL), and intermediate 11 and diethyl squalate (0.12 mL, 0.84 mmol) were added. This mixture was stirred at RT for 3 hours and then concentrated. The crude product was purified by flash column chromatography (reverse phase, C18) under a gradient of MeCN (10%~80%) in water (0.1% v / v HCOOH) to obtain intermediate 71,3-ethoxy-4-[3-ethyl-5-(1H-tetrazole-5-yl)anilino]cyclobuta-3-en-1,2-dione (203 mg, 0.65 mmol, yield 77%) as a cream-colored solid. The data can be viewed in Table 2. 【0206】 Typical synthesis procedure in the examples Route A Example 2: Preparation procedure for 3-((3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione 【0207】 [ka] Step (i): To a solution of intermediate 72, 3-amino-5-(trifluoromethyl)benzonitrile (150 mg, 0.81 mmol) in DMF (3 mL), NaN3 (65 mg, 1.62 mmol) and NH4Cl (53.5 mg, 1.62 mmol) were added. The mixture was heated to 130 °C for 20 hours, then cooled to RT and acidified to pH 1 with 1 M HCl (aq). The mixture was extracted with siRNA to remove organic compounds, washed with water and brine, and dried by passing through hydrophobic frit to concentrate. The crude product was purified by reverse-phase flash chromatography (C18 silica) under a gradient of MeCN (0-50%) in denatured water (containing 0.1% v / v HCOOH) to obtain the product intermediate 73, 3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)aniline (144 mg, 0.63 mmol, yield 77%). The data can be viewed in Table 2. 【0208】 Step (ii): Intermediate 73, 3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)aniline (77 mg, 0.34 mmol) was dissolved in EtOH (1.4 mL), to which intermediate 11, diethyl squalate (0.05 mL, 0.34 mmol), and TEA (0.05 mL, 0.38 mmol) were added. The mixture was stirred at RT for 1 hour, then concentrated to dryness. The crude product was purified by reverse-phase flash chromatography (C18 silica) under a gradient of MeCN (0-60%) in denatured water (0.1% v / v HCOOH) to obtain the product, intermediate 74, 3-((3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)phenyl)amino)-4-ethoxycyclobuta-3-en-1,2-dione (51.6 mg, 0.15 mmol, yield 44%). The data can be viewed in Table 2. 【0209】 Step (iii): Intermediate 74, 3-((3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)phenyl)amino)-4-ethoxycyclobuta-3-en-1,2-dione (39 mg, 0.11 mmol) was dissolved in a 10:1 mixture of THF and 1 M HCl (aq) (0.55 mL), heated to 60°C for 3 hours, and then concentrated. The crude product was purified by preparative HPLC [reverse phase (Kinetex C18, 100 × 30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: solvent B 5%~95% (over 10 minutes), solvent B 100% (over 2 minutes), solvent A: 0.1% TFA water. Solvent B: MeCN] to obtain Example 2, 3-((3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione (15.3 mg, 0.047 mmol, yield 43%). The data can be viewed in Table 3. 【0210】 Route B Example 5: Preparation procedure for N-benzyl-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide 【0211】 [ka] Step (i): Intermediate 12, a suspension of N-benzyl-3'-((2-ethoxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (1.43 g, 2.89 mmol) in THF (15 mL) and 1 M HCl (aq) (1.5 mL) was heated to 60 °C for 24 hours, after which the mixture was concentrated. The crude product was purified by preparative HPLC [reverse phase (Gemini-NX C18, 100×30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: solvent B 5%~35% (over 10 minutes), solvent B 100% (over 2 minutes), solvent A: 0.2% v / v 28% ammonia-containing aqueous solution. Solvent B: MeCN]. The obtained solid was suspended in 1 M HCl (aq) and vigorously stirred for 2 hours, then recovered by filtration. The wet solid was suspended in toluene and concentrated to obtain Example 5, N-benzyl-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (720 mg, 1.55 mmol, yield 53%) as a pale green solid. The data can be viewed in Table 3. 【0212】 Route C Example 8: Preparation procedure for 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(4-(2-methoxyethoxy)benzyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide 【0213】 [ka] Step (i): Intermediate 29, 3'-((2-ethoxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxylic acid (60 mg, 0.15 mmol) and DIPEA (0.08 mL) were dissolved in DMF (0.7 mL), to which HATU (68.4 mg, 0.18 mmol) was added, followed by intermediate 75, (4-(2-methoxyethoxy)phenyl)methanamine (29.5 mg, 0.17 mmol). The mixture was stirred at RT for 1 hour, then diluted with DMSO and purified by preparative HPLC [reverse phase (Kinetex C18, 100×30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: solvent B 40%~70% (over 10 minutes), solvent B 100% (over 2 minutes), solvent A: water containing 0.1% TFA. Solvent B: MeCN] to obtain intermediate 76, 3'-((2-ethoxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(4-(2-methoxyethoxy)benzyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (36.3 mg, 0.064 mmol, yield 43%) as a pale green solid. The data can be viewed in Table 2. 【0214】 Step (ii): Intermediate 76, a suspension of 3'-((2-ethoxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(4-(2-methoxyethoxy)benzyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (36.3 mg, 0.064 mmol) in THF (1 mL) and 1 M HCl (aq) (0.1 mL) was heated to 60°C for 13 hours, then concentrated to dryness. The crude product was purified by preparative HPLC [reverse phase (Kinetex C18, 100 × 30 mm, 5 μm, 30 mL / min, gradient of solvent B in solvent A: solvent B 40%~30% (over 10 minutes), solvent B 100% (over 2 minutes), solvent A: water containing 0.1% TFA. Solvent B: MeCN] to obtain Example 8, 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(4-(2-methoxyethoxy)benzyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (16 mg, 0.03 mmol, yield 46%) as a pale green solid. The data can be viewed in Table 3. 【0215】 Route D Example 15: Preparation procedure for 3-((5-(1H-tetrazole-5-yl)-4'-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)amino)-4-hydroxycyclobuta-3-ene-1,2-dione 【0216】 [ka] Step (i): Intermediate 31, 3-((3-bromo-5-(1H-tetrazole-5-yl)phenyl)amino)-4-ethoxycyclobuta-3-ene-1,2-dione (0.50 g, 1.37 mmol), intermediate 77, 4,4,5,5-tetramethyl-2-(4-(trifluoromethyl)phenyl)-1,3,2-dioxaborolane (0.56 g, 2.06 mmol), and K2CO3 (0.38 g, 2.75 mmol) were suspended in MeCN (5 mL) and water (5 mL) and degassed with N2 for 15 minutes. PdCl2 (dtbpf) (0.089 g, 0.13 mmol) was added, and the mixture was heated to 100°C for 1 hour under microwave irradiation. After this time, the solvent was removed under reduced pressure, and the crude product was purified by preparative HPLC [reverse phase (X-bridge C18, 250 × 30 mm, 5 μm, 27 mL / min, gradient of solvent B in solvent A: solvent B 10%~98% (over 59 min), solvent B 10% (over 2 min), solvent A: 10 mM NH4HCO3 water. Solvent B: MeCN] to obtain Example 15, 3-((5-(1H-tetrazole-5-yl)-4'-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)amino)-4-hydroxycyclobuta-3-en-1,2-dione (0.35 g, 0.87 mmol, yield 64%) as a brown solid. The data can be viewed in Table 3. 【0217】 Route E Example 17: Preparation procedure for 3-hydroxy-4-((6-methoxy-4-(1H-tetrazole-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione 【0218】 [ka] Step (i): To an ice-cold suspension of intermediate 78, 6-methoxy-4-(1H-tetrazole-5-yl)pyridine-2-amine (510 mg, 2.56 mmol) in EtOH (10 mL), TEA (1.00 mL, 7.69 mmol) was added. The mixture was stirred at the same temperature for 15 minutes, then intermediate 11, diethyl squalate (479 mg, 2.82 mmol), was added dropwise, and the reaction was stirred at RT for 16 hours. The mixture was concentrated, and the residue was partitioned between ethyl acetate and water. The organic compounds were removed, and the pH of the aq. layer was acidified to 1-2 with 1N aq.HCl, and the mixture was stirred for 30 minutes. The obtained precipitate was collected by filtration, washed with ice-cold water, and dried under high vacuum to obtain the intermediate 79,3-ethoxy-4-((6-methoxy-4-(1H-tetrazole-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione (350 mg, 1.11 mmol, yield 43%) as a yellow solid. The data can be viewed in Table 2. 【0219】 Step (ii): Intermediate 79, a suspension of 3-ethoxy-4-((6-methoxy-4-(1H-tetrazole-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione (350 mg, 1.11 mmol) in THF (5.5 mL) and 1N aq.HCl (0.55 mL) was heated to 60°C for 24 hours, and the mixture was concentrated to dryness. The residue was triturated from  to obtain Example 17, 3-hydroxy-4-((6-methoxy-4-(1H-tetrazole-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione (170 mg, 0.59 mmol, yield 53%) as a yellow solid. The data can be viewed in Table 3. 【0220】 Root F Example 30: Preparation procedure for 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N,N-dimethyl-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide 【0221】 [ka] Step (i): Intermediate 1,3-amino-5-bromobenzonitrile (500 mg, 2.53 mmol), intermediate 80, (4-(dimethylcarbamoyl)phenyl)boronic acid (587 mg, 3.04 mmol), K2CO3 (700 mg, 5.07 mmol), and Pd(Ph3P)4 (59 mg, 0.050 mmol) were heated to 90°C (8 mL) in 1,4-dioxane (8 mL) and water (2 mL) for 16 hours. The mixture was cooled and partitioned between pharmaceutically acceptable and water. The organic compounds were separated, and the aqueous layer was washed with pharmaceutically acceptable. The combined organic compounds were dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography (conventional phase, silica) under a gradient of SiO(0%) to hexane to obtain intermediate 81,3'-amino-5'-cyano-N,N-dimethyl-[1,1'-biphenyl]-4-carboxamide (210 mg, 0.79 mmol, yield 31%) as brown gum. The data can be viewed in Table 2. 【0222】 Step (ii): Intermediate 81, a solution of 3'-amino-5'-cyano-N,N-dimethyl-[1,1'-biphenyl]-4-carboxamide (200 mg, 0.754 mmol), NaN3 (79.9 mg, 1.50 mmol), and NH4Cl (98 mg, 1.50 mmol) in DMF (2 mL) was heated to 130 °C for 16 hours. The mixture was cooled and the pH was acidified to 1 using 1 M HCl (aq). The obtained precipitate was collected by filtration and purified by preparative HPLC [reverse phase (Sunfire C18, 250×19mm, 5μm, 15mL / min, gradient of solvent B in solvent A: solvent B 8%~25% (over 22 min), solvent B 22~25% (over 5 min), solvent B 100% (over 2 min), solvent B 100~8% (over 3 min), solvent A: 0.1% HCOOH water. Solvent B: MeCN] to obtain intermediate 82, 3'-amino-N,N-dimethyl-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (22 mg, 0.071 mmol, yield 9%) as a white solid. The data can be viewed in Table 2. 【0223】 Step (iii): A solution of intermediate 82, 3'-amino-N,N-dimethyl-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (22 mg, 0.071 mmol) and intermediate 83, squalate (8.1 mg, 0.071 mmol) in water (1 mL) was heated to 125°C for 6 hours in a sealed tube. The mixture was cooled, and the solid was collected by filtration to obtain Example 30, 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N,N-dimethyl-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide (10 mg, 0.025 mmol, yield 35%) as a yellow solid. The data can be viewed in Table 3. 【0224】 [Examples] The present invention will be described hereafter with reference to the following embodiments, but is not limited thereto. 【0225】 [Examples 1-31] The compounds of Examples 1 to 31, shown in Table 1 below, were prepared. Their NMR and LCMS properties, as well as the methods used to prepare them, are shown in Table 3. The starting materials for each example are listed in Table 2. 【0226】 [Table 1] TIFF0007875883000060.tif255166TIFF0007875883000061.tif198169 【0227】 [Table 2] TIFF0007875883000063.tif255157TIFF0007875883000064.tif255157TIFF0007875883000065.tif255157TIFF0007875883 000066.tif255157TIFF0007875883000067.tif255158TIFF0007875883000068.tif255158TIFF0007875883000069.tif94165 【0228】 [Table 3] TIFF0007875883000071.tif243170TIFF0007875883000072.tif243170TIFF0007875883000073.tif243170 TIFF0007875883000074.tif246170TIFF0007875883000075.tif241170TIFF0007875883000076.tif225170 【0229】 crystalline compound Preparation of the tromethamine salt (hydrate I) (unseeded) of compound A. 【0230】 [ka] Compound A is the compound of Example 5 (structure shown above). 2 mL of 97.6% (THF:water 3:1):2.4% DMSO was added to 50 mg of Compound A, and the mixture was heated to 50°C to dissolve. 1 equivalent of 1 M tromethamine aqueous solution was added to the solution, and it was equilibrated at 50°C for 1 hour before cooling to room temperature, with stirring overnight. The solution was then evaporated under nitrogen (to approximately 25% of its original volume) until a precipitate formed. The resulting solid was filtered, washed with IPA, and dried under vacuum at 45°C. 【0231】 Preparation of the tromethamine salt (hydrate I) (seed) of compound A. Compound A (1 kg scale) was placed in a container under nitrogen. Subsequently, THF (6.08 vol) and an aqueous solution of tromethamine (0.268 wt tromethamine dissolved in 2.21 vol of water) were added. Then water (1.84 vol) was added to the container. The mixture was heated and dissolved at 60°C. The solution was cooled to 50°C. Acetonitrile (1.29 vol) was added before the reaction was seeded with crystalline hydrate I of the tromethamine salt of compound A (0.0126 wt). The mixture was equilibrated for 1 hour. Then acetonitrile (6.33 vol) was added to the slurry over 2 hours. The mixture was then cooled to 5°C and stirred overnight. The solid was washed with acetonitrile (3.96 vol) before drying under vacuum at 45°C to obtain tromethamine salt (hydrate I) of compound A. 【0232】 Properties of the tromethamine salt (hydrate I) of compound A The X-ray powder diffraction (XRPD) pattern of hydrate I of the tromethamine salt of compound A is shown in Figure 1, and the diffraction angles and interplanar spacing d are summarized in Table 4 (characteristic peaks) and Table 5 (complete peak list). XRPD analysis was performed using a PANalytical Xpert Pro diffractometer equipped with Si zero-background wafers. The acquisition conditions were CuK α The parameters included radiation, generator voltage: 40kV, generator current: 45mA, step width: 0.02°2θ, start angle: 2.0°2θ, and end angle: 40.0°2θ. 【0233】 [Table 4] 【0234】 [Table 5] 【0235】 An example of a differential scanning calorimeter (DSC) thermogram of hydrate I of the tromethamine salt of compound A is shown in Figure 2. DSC analysis was performed using a TA Instruments Q100 differential scanning calorimeter equipped with an autosampler and a refrigeration system under a 50 mL / min N2 purge. DSC thermograms of the samples were acquired at 10°C / min in a crimped Al pan. The DSC thermogram shows endothermic activity at an initiation temperature of approximately 155°C; however, this can vary depending on experimental conditions and crystallinity levels. 【0236】 An example of a thermogravimetric analysis (TGA) thermogram of hydrate I, the tromethamine salt of compound A, is shown in Figure 3. TGA analysis was performed using a TA Instruments Q5000 thermogravimetric analyzer at a heating rate of 10°C / min under an N2 flow at 25 mL / min. The TGA thermogram of this hydrate typically shows a weight loss between 6 and 7% at 30 and 120°C, which corresponds to approximately 2 to 2.5 equivalents of water in each equivalent of compound A, i.e., the variable hydrate. 【0237】 Preparation of the tromethamine salt (hydrate II) (unseeded) of compound A. 2 mL of methanol was added to 100 mg of the amorphous form of the tromethamine salt of compound A, and the suspension was allowed to equilibrate overnight at room temperature. The resulting solid was filtered and dried under vacuum at 45°C. 【0238】 Preparation of the tromethamine salt (hydrate II) (seed) of compound A. 3 g of hydrate I of the tromethamine salt of compound A was added to a container with 60 mL of methanol while overhead stirring. 60 mg (with 2% w / w seed added) of hydrate II of the tromethamine salt of compound A was added, and the reaction mixture was allowed to equilibrate overnight at room temperature. The suspension thickened and its color faded overnight. The resulting solid was filtered and dried at 45°C under vacuum. 【0239】 The X-ray powder diffraction (XRPD) pattern of hydrate II of the tromethamine salt of compound A is shown in Figure 4, and the diffraction angles and interplanar spacing d are summarized in Table 6 (characteristic peaks) and Table 7 (complete peak list). XRPD analysis was performed using a PANalytical Xpert Pro diffractometer equipped with a Si-zero background wafer. The acquisition conditions were CuK α The parameters included radiation, generator voltage: 45kV, generator current: 40mA, step width: 0.03°2θ, start angle: 3.0°2θ, and end angle: 35.0°2θ. 【0240】 [Table 6] 【0241】 [Table 7] 【0242】 An example of a differential scanning calorimeter (DSC) thermogram of hydrate II of the tromethamine salt of compound A is shown in Figure 5. DSC analysis was performed using a PerkinElmer Pyris 6000 differential scanning calorimeter equipped with an autosampler and refrigeration system under a 20 mL / min N2 purge. DSC thermograms of the samples were acquired at 20°C / min in a pinhole Al pan. The DSC thermogram shows endothermic activity at an initiation temperature of approximately 210°C; however, this may vary depending on experimental conditions and crystallinity levels. 【0243】 An example of a thermogravimetric analysis (TGA) thermogram of hydrate II of the tromethamine salt of compound A is shown in Figure 6. TGA analysis was performed using a PerkinElmer Pyris 1 thermogravimetric analyzer at a heating rate of 20°C / min under an N2 flow at 20 mL / min. The TGA thermogram of this hydrate typically shows a weight loss between 1 and 3% at 30 and 150°C, which corresponds to approximately 0.3 to 1 equivalent of water in each equivalent of compound A, i.e., the variable hydrate. 【0244】 To determine the critical water activity at which hydrate I and hydrate II are stable, water activity studies were conducted. Slurries with high prominence of hydrate I and hydrate II were tested at room temperature within a range of aqueous solvent mixtures with varying water activity. Hydrate I was isolated from all mixtures with a water activity greater than 0.5. The equilibrium of hydrate II alone supported its conversion to hydrate I in solvent mixtures with a water activity of 0.5 or higher. A slurry with high prominence of hydrate I and hydrate II in a solvent mixture with a water activity of 0.2 at room temperature yielded a mixture of solid hydrate I and hydrate II. 【0245】 Preparation of crystalline compound A (free acid) (Pattern 3) Crude compound A (free acid) was purified by reverse-phase chromatography under basic conditions (high pH) under a 5-35% acetonitrile gradient in an aqueous medium (0.2% 28% ammonium hydroxide aqueous solution) using a Gilson Semi Preparative HPLC, Pumps 332 & 331, GX-271 Liquid handler, and Trilution software, with a flow rate of 30 mL / min and 171 diode array detectors at 205 nm, 210 nm, and 230 nm. The chromatography was performed over 12.5 minutes via a Gemini-NX C18 column (5 μm, 100 × 30 mm). After combining the desired fractions, the mixture was evaporated using a Biotage V10 apparatus to obtain a white solid residue (4 g) and a diammonium salt. 【0246】 Diammonium salt (4 g, 7.99 mmol) was dissolved in DMSO (39.96 mL) and stirred for 30 minutes. 1N HCl (59.94 mL, 59.94 mmol) was added, the resulting precipitate was collected by filtration, washed with ice-cold water (20 mL), and dried to obtain the crude product. This crude product was resuspended in EtOH (40 mL) and stirred for 3 hours. The suspension was then filtered to obtain a dry white solid, which was ground into a fine powder (2.94 g). NMR confirmed that this powder was the desired product, but a large amount of DMSO remained in the sample. Therefore, the solid was resuspended in EtOH (25 mL) and stirred for 18 hours. The suspension was then filtered, the solid was collected, and ground into a fine powder using a mortar and pestle to obtain the free acid of compound A (2.39 g, 5.12 mmol, yield 64.1%) as a white crystalline solid. The NMR spectrum of the product shows that the ratio of product to DMSO was approximately 1:0.05, indicating that a small amount of DMSO remained in the sample. The X-ray powder diffraction (XRPD) pattern of the free acid of crystalline compound A is shown in Figure 7, where this crystalline solid is identified as "Pattern 3". 【0247】 An example of a thermomass spectrometry (TGA) thermogram of free acid pattern 3 for compound A is shown in Figure 8. 【0248】 Preparation of crystalline compound A (free acid) (Pattern 1) When the free acid pattern 3 of crystalline compound A was dried by heating overnight at 45°C under vacuum (approximately 10-15 mbar), a different crystalline solid referred to as "pattern 1" was obtained. The XRPD pattern of free acid pattern 1 of compound A is shown in Figure 9. 【0249】 An example of a thermomass spectrometry (TGA) thermogram of free acid pattern 1 of compound A is shown in Figure 10. 【0250】 Physiological activity [Example A] Human GPR35a isoform conjugation 2.5 × 10 6Overexpression of human GPR35a baculovirus in HEK293f cells at a cell density of 10 cells / mL and an infection multiplicity of 2.5 over 24 hours in Pro293(Lonza) + 5% FBS, 1% Glutamax, and 0.4% Pen / Strep. Cells were harvested and centrifuged at 2500 RPM for 10 minutes at 4°C. The supernatant was discarded 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). Homogenization was performed in a mechanical homogenizer (VMR) for 10 seconds. The membrane was centrifuged in a centrifuge tube at 40,000 g at 4°C for 15 minutes. The supernatant was discarded and the cell was resuspended in 15 mL of homogenization buffer. Homogenization was performed for 20 seconds. The membrane was centrifuged at 40,000 g at 4°C for 45 minutes. The membrane was resuspended in 3 mL of storage buffer (20 mM HEPES, 0.1 mM EDTA, pH 7.4) while mixing thoroughly. The resulting membrane was then stored at -80°C. 【0251】 GPR35 cell membrane homogeneous material was resuspended in binding buffer (50 mM TRIS + 10 mM MgCl2 pH 7.4) at the final assay concentration of 5 ug / well. A 10-point 1 / 2 log concentration curve was formed by diluting the test compound in dimethyl sulfoxide (DMSO (Sigma Aldrich, UK)). The test compound was added to each plate, followed by 7 nM 3H-27966. 【0252】 To calculate non-specific binding, 0.1 μM FAC rhodoxamide was added. Finally, the membrane was added to each well on the plate. After incubation at room temperature for 60 minutes, the membrane was filtered onto a 96-well white microplate joined with Unifilter and GF / B filters, pre-immersed in ddH2O using a TomTec cell harvester, and washed five times with distilled water. The plate was dried and sealed before the addition of 50 μl / well scintillant, and radioactivity was measured using a MicroBeta analyzer. The IC50 value was derived from the inhibition curve, and the affinity constant (Ki) value was derived from the Chen-Prusov formula below: pKi = -log10Ki It was calculated using 【0253】 Label-free DMR functional assay HT-29 cells (ATCCHTB-38) were stored in continuous culture in McCoys (Thermo 16600082) supplemented with 10% FBS. The day before the assay, cells were harvested using TrypLE (Gibco 12604-013) and seeded overnight at 37°C, 5% CO2, in Corning EPIC 384-well plates (5040) in a total volume of 50 μl of medium at 20 kJ / well. On the day of the assay, the cell medium was removed and replaced with assay buffer (HBSS + 20 mM HEPES pH 7.4), and the plates were re-incubated for 1 hour. The compound was prepared in 100% DMSO in an ECHO LDV384 source plate. The cell plate was read using an EPIC plate reader at room temperature for 15 minutes, then the reading was interrupted, and the cell plate was placed in a LabCyte ECHO 550 to add 50 nl of the compound per well by acoustic transfer. The plate was immediately returned to the EPIC reader, reading was resumed, and dynamic mass redistribution was measured for 60 minutes. The raw data was analyzed using EPIC analyzer software, and the maximum peak per concentration was obtained to measure EC50. All raw DMR data were normalized to rhodoxamide and buffer corrected. 【0254】 [Table 8] 【0255】 Ki values ​​were derived from concentration-response analysis in HEKf cell membranes overexpressing the GPR35a isoform. EC50 values ​​were derived from concentration-response analysis in HT-29 cell lines endogenously expressing human GPR35. 【0256】 Compound A demonstrated approximately 500-fold higher functional capacity of GPR35 than cromolyn, a mast cell stabilizer that has been clinically used at high doses for GI disorders. Compound A also showed pharmacological effects in PGE2-induced fluid secretion, indomethacin-induced ileitis and barrier permeability, a TNBS mouse visceral pain model, and in preclinical species including acute LPS challenges in rats and mice. Compound A further demonstrated strong selectivity for GPR35, and no off-target effects were observed. Compound A has very low potential drug interactions with major human CYPs, including CYP3A4.

Claims

[Claim 1] Compound of formula (1) 【Chemistry 1】 or its salt or its tautomer (In the formula, X is either N or CH. R 1 H or halo, R 2 H is H, halo; (i) C, which is arbitrarily substituted with 1 to 6 fluorine atoms. 1-6 Alkyl, C optionally substituted with 1 to 6 fluorine atoms 3-6 Cycloalkyl or C optionally substituted with 1 to 6 fluorine atoms 1-6 Alkoxy; or (ii) an optionally substituted aryl, optionally substituted O-aryl, optionally substituted heteroaryl, optionally substituted monocyclic heteroaryl, or optionally substituted bicyclic heteroaryl, where the optionally selected substituent is R 3 And, Here, R 3 is H, halo, C optionally substituted with 1 to 6 fluorine atoms 1-6 alkyl, C optionally substituted with 1 to 6 fluorine atoms 3-6 cycloalkyl, C optionally substituted with 1 to 6 fluorine atoms 1-6 alkoxy, -CO 2 R 4 , -CONHCH 2 R 4 , -CONHCH 2 CH 2 OR 4 , -OR 4 , -OCH 2 R 4 , -CH 2 R 4 , -OCH 2 R 4 , -CH 2 CH 2 OR 4 , -OCH 2 CH 2 OR 4 , -CONHR 4 or -CON(CH 3 )R 4 ; Here, R 4 This is C, which is optionally substituted with H and 1 to 6 fluorine atoms. 1-6 Alkyl, or the following groups: 【Chemistry 2】 {During the ceremony, R 5 , R 6 and R 7 However, independently H, halo, CO 2 R 8 CONR 8 R 9 Or C which is arbitrarily substituted with 1 to 6 fluorine atoms 1-6 It is alkyl, and here C 1-6 One or two carbon atoms of the alkyl group may be optionally substituted with oxygen; R 8 and R 9 C is independently substituted with H or 1 to 6 fluorine atoms as desired. 1-6 It is alkyl. (That is the case.) [Claim 2] The compound according to claim 1, or a salt thereof or a tautomer thereof, wherein X is CH. [Claim 3] R 1 The compound according to claim 1, or a salt thereof or a tautomer thereof, wherein is H. [Claim 4] R 1 The compound according to claim 1, or a salt thereof or a tautomer thereof, wherein is Cl or F. [Claim 5] R 2 C is substituted with H, and any one to six fluorine atoms. 1~6 Alkyl, C substituted with any one to six fluorine atoms 3~6 Cycloalkyl, or C substituted with any one to six fluorine atoms 1~6 The compound according to claim 1, or a salt thereof or a tautomer thereof, which is an alkoxy compound. [Claim 6] R 2 However, R can be arbitrarily selected. 3 Phenyl substituted with, optionally R 3 Pyridyl is substituted with, optionally R 3 O-phenyl substituted with, optionally R 3 Indazolyl substituted with, or optionally R 3 The compound according to claim 1, or a salt thereof or a tautomer thereof, wherein the pyridadinyl is substituted with [the specified compound]. [Claim 7] R 3 OMe, CO 2 H, CO 2 Et, CON(CH 3 ) 2 CONHCH 2 CH 2 OCH 3 , or 【Transformation 3】 A compound according to claim 6, or a salt thereof or a tautomer thereof, selected from the group consisting of the above. [Claim 8] Compounds of formula (3a), (3b), (3c), (3d), or (3e) 【Chemistry 4】 The compound according to claim 6, or a salt thereof or a tautomer thereof. [Claim 9] Compounds of formula (4a), (4b), (4c), (4d), or (4e) 【Transformation 5】 The compound according to claim 6, or a salt thereof or a tautomer thereof. [Claim 10] R 2 but, 【Chemistry 6-1】 【Chemistry 6-2】 A compound according to claim 1, or a salt thereof or a tautomer thereof, selected from the group consisting of the above. [Claim 11] a) X is CH b)R 1 is H; and c)R 2 is an optionally substituted aryl, optionally substituted O-aryl, optionally substituted heteroaryl, optionally substituted monocyclic heteroaryl, or optionally substituted bicyclic heteroaryl, where the optional substituent is R 3 The compound according to claim 1, or a salt thereof or a tautomer thereof. [Claim 12] 3-((3-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(trifluoromethyl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((5-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-yl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; Ethyl 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazol-5-yl)-[1,1'-biphenyl]-3-carboxylate; N-benzyl-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; N-benzyl-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-carboxamide; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(2-methoxyethyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N-(4-(2-methoxyethoxy)benzyl)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; 3-Hydroxy-4-((3'-Methoxy-5-(1H-Tetazol-5-yl)-[1,1'-Biphenyl]-3-yl)amino)cyclobuta-3-en-1,2-dione; 3-((3-ethyl-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-3-carboxylic acid; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-N-(4-(trifluoromethyl)benzyl)-[1,1'-biphenyl]-4-carboxamide; 3-((4-fluoro-3-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((4-chloro-3-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((5-(1H-tetrazole-5-yl)-4'-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(6-(benzyloxy)pyridine-3-yl)-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((6-Methoxy-4-(1H-Tetazol-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((6-phenyl-4-(1H-tetrazole-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazol-5-yl)-5-(2-(4-(trifluoromethyl)benzyl)-2H-indazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(6-((4-(trifluoromethyl)benzyl)oxy)pyridine-3-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(2-benzyl-2H-indazole-5-yl)-5-(1H-tetrazol-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((6-Phenoxy-4-(1H-Tetazol-5-yl)pyridine-2-yl)amino)cyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(2-(2-(4-(trifluoromethyl)phenoxy)ethyl)-2H-indazole-6-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-cyclopropyl-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-indazole-5-yl)-5-(1H-tetrazol-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(6-(2-(3,5-bis(trifluoromethyl)phenoxy)ethoxy)pyridine-3-yl)-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-(1H-tetrazole-5-yl)-5-(6-(2-(4-(trifluoromethyl)phenoxy)ethoxy)pyridine-3-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-((3-cyclohexyl-5-(1H-tetrazole-5-yl)phenyl)amino)-4-hydroxycyclobuta-3-en-1,2-dione; 3-Hydroxy-4-((3-(pyridazin-4-yl)-5-(1H-tetrazole-5-yl)phenyl)amino)cyclobuta-3-en-1,2-dione; 3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-N,N-dimethyl-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide; N-(4-carbamoylbenzyl)-3'-((2-hydroxy-3,4-dioxocyclobuta-1-en-1-yl)amino)-5'-(1H-tetrazole-5-yl)-[1,1'-biphenyl]-4-carboxamide A compound according to claim 1, or a salt thereof or a tautomer thereof, selected from the group consisting of the above. [Claim 13] A salt according to any one of claims 1 to 12, which is a pharmaceutically acceptable salt. [Claim 14] The compound, 【Transformation 7】 The compound or salt according to claim 1, or a pharmaceutically acceptable salt thereof. [Claim 15] The compound, 【Transformation 8】 The compound according to claim 1. [Claim 16] Structure below: 【Chemistry 9】 A pharmaceutically acceptable salt according to claim 15, which is a trometamine salt having the following properties. [Claim 17] A pharmaceutical composition comprising the compound or salt described in claim 1, and a pharmaceutically acceptable excipient. [Claim 18] i) Treatment of mast cell disorders, acute and chronic pain conditions, or diseases associated with allergic or inflammatory disorders of both the gastrointestinal tract and the lungs; or ii) Treatment of gastrointestinal disorders or conditions, pain symptoms related to gastrointestinal diseases or conditions of other internal organs, or lung diseases or conditions. The composition according to claim 17 for use in [the specified purpose]. [Claim 19] i) The gastrointestinal disorder or condition is selected from the group consisting of food allergies, food intolerances and allergic disorders, celiac disease, systemic mast cell disease and other mast cell-related disorders (mast cell activation syndrome, clonal mast cell dystrophy, monoclonal mast cell activation syndrome, idiopathic urticaria, idiopathic anaphylaxis), mast cell colitis, irritable bowel syndrome (IBS), gastrointestinal motility disorders, functional gastrointestinal disorders, gastroesophageal reflux disease (GERD), duodenal reflux, diarrheal diseases, eosinophilic gastroenteritis, eosinophilic esophagitis, infectious diarrhea (e.g., Clostridium difficile, Salmonella, Shigella toxin), microscopic colitis, immune-mediated gastrointestinal disorders, Crohn's disease, ulcerative colitis, inflammatory bowel disease, and visceral abdominal pain, or ii) The composition for use according to claim 18, wherein the pain symptoms are related to a gastrointestinal disease or other visceral disease selected from the group consisting of Crohn's disease, ulcerative colitis, inflammatory bowel disease, radiation colitis, radiation cystitis, celiac disease, glutenic enteropathy, radiation cystitis, interstitial cystitis, bladder pain syndrome, cancer, gastroesophageal reflux disease, chemotherapy mucositis and radiotherapy mucositis, pancreatitis, prostatitis, pelvic pain, endometriosis, hepatitis, hepatic fibrosis and cirrhosis. [Claim 20] The composition for use according to claim 18, wherein the lung disease or condition is selected from the group consisting of chronic obstructive pulmonary disease, asthma, chronic bronchitis, cystic fibrosis, emphysema, chronic idiopathic cough, hypersensitivity respiratory disease, and idiopathic pulmonary fibrosis. [Claim 21] A pharmaceutically acceptable salt crystal according to claim 16. [Claim 22] The crystal according to claim 21, which is a hydrate. [Claim 23] CuK α The crystal according to claim 22, having an XRPD pattern with diffraction angles of 3.9±0.2, 7.7±0.2, 10.0±0.2, and 15.8±0.2°2θ when measured using radiation. [Claim 24] CuK α The crystal according to claim 22, having an XRPD pattern with diffraction angles of 4.4±0.2, 14.4±0.2, and 23.9±0.2°2θ when measured using radiation.

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