Prevention, inhibition, or treatment of conditions or disorders mediated by urocanate reductase.

Abstract

The present invention provides a compound of formula (I), a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof for use in preventing, inhibiting, or treating a condition or disorder mediated by urocanic acid reductase, wherein A is a 5-membered heteroaromatic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, at least one of the heteroatoms being nitrogen, R 1 and R 2 are independently selected from -H, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkoxy, halogen, and -CN, and when present, each R 3 is C 1-6 alkyl, C 1-6 alkoxy, C 1-3 alkoxy, halogen, -OH, -CN, -NO2, -NR 5 R 6 (wherein R 5 and R 6 are independently selected from H and C 1-3 alkyl), and -C(=NR 7 )(NR 8 R 9 (wherein each of R 7 , R 8 , and R 9 is independently selected from H and C 1-3 alkyl), and R 4 is selected from -OH, -SH, C 1-6 alkoxy, and -NR 10 R 11 (wherein R 10 and R 11 are independently selected from H and C 1-3 alkyl), and n is an integer from 0 to 2. Such compounds are particularly suitable for use in preventing, inhibiting, or treating gastrointestinal disorders or conditions, cardiovascular diseases or disorders, and metabolic disorders, such as, but not limited to, primary sclerosing cholangitis (PSC), cardiomyopathy, heart failure, type 2 diabetes, and prediabetes. The present invention further provides certain novel compounds, methods for their preparation, and pharmaceutical compositions containing such compounds. JPEG2026525257000099.jpg25128

Description

[Technical Field]

[0001] The present invention relates to the use of compounds in preventing, inhibiting, or treating conditions mediated by urocanate reductase, more specifically, conditions mediated by urocanate reductase present in a target intestinal microbiota. In particular, the present invention relates to the use of such compounds in preventing, inhibiting, or treating conditions resulting from and / or related to the production of imidazole propionates, such as metabolic disorders, cardiovascular disorders, and gastrointestinal disorders. The present invention further relates to certain novel compounds, pharmaceutical compositions containing them, and their use in such treatments. [Background technology]

[0002] Urocanic acid reductase is a bacterial enzyme that catalyzes the conversion of urocanic acid to imidazole propionate (ImP) in the microbial histidine degradation pathway.

[0003] Elevated ImP levels are associated with a wide range of conditions, diseases, and disorders. These include metabolic disorders such as type 2 diabetes (T2D), various cardiovascular disorders such as cardiovascular disease (CVD), heart failure (HF), and myocardial fibrosis, and various gastrointestinal conditions such as inflammatory bowel disease (IBD).

[0004] Metabolic disorders may be associated with changes in the structure and function of the gut microbiota. The microbial metabolite ImP is present at higher concentrations in individuals with T2D and impaired glucose tolerance (IGT) compared to healthy individuals with normal glucose tolerance (NGT) (Koh et al., Cell 175, 947-961, e17, 2018; and Molinaro et al., Nat. Commun. 11, 5881, 2020). Microbial-produced ImP reduces insulin receptor substrate (IRS) protein levels and impairs insulin signaling via the p38γ mitogen-activated protein kinase (MAPK) activation pathway, which is a surrogate target of the mechanistic target of lapacimin complex 1 (mTORC1). Therefore, ImP contributes to the pathogenesis of T2D and IGT. Furthermore, elevated ImP levels are also associated with the fact that metformin is unable to lower blood glucose levels by interfering with AMPK phosphorylation via the p38γ / Akt-dependent pathway (Koh et al., Cell Metabolism 32(4):643-653, 2020).

[0005] Plasma ImP concentration has been shown to be positively correlated with diastolic blood pressure (Son et al., Nutrients 13, 2021). Pharmacological inhibition of p38γ, a molecular target of ImP, has also been found to reduce myocardial fibrosis in patients with HFpEF (Lewis et al., Nature Medicine 27, 1477-1482, 2021), and activation of p38γ / δ has been shown to induce cardiac hypertrophy and therefore potentially contribute to diabetic cardiomyopathy (DCM) (Gonzalez-Teraen et al., Nat.Commun. 7:10477, 2016). More recently, it has been demonstrated that ImP levels are associated with CVD and HF in human subjects, independently of T2D and other established cardiovascular risk factors, and that ImP levels are an independent risk factor for overall mortality (Molinaro et al., JACC Heart Fail. April 1, 2023; S2213-1779(23)00138-5, doi:10.1016 / j.jchf.2023.03.008). Specifically, elevated ImP levels were found to be associated with decreased left ventricular systolic ejection fraction and as a predictor of 5-year mortality. Therefore, increased levels of the intestinal microbial metabolite ImP contribute to CVD and HF and are a predictor of overall survival.

[0006] Early studies reported that ImP is excreted from patients with bowel disorders but is hardly present in the feces and urine of healthy subjects (Van Der Heiden et al., Clinica Chimica Acta 39, 201-214, 1972). ImP is associated with inflammatory bowel disease, and rectal ImP administration has been shown to induce intestinal inflammation, impair the intestinal barrier, and affect goblet cell proliferation (Vich Vila et al., Gut 0:1-14, 2023; and Wu et al., Mol. Nutr. Food Res. 66, e2101175, 2022). ImP is also associated with colorectal cancer (Gao et al., Gastroenterology 163(4):P1024-1037, 2022). Previous studies have shown that the mTORC pathway, including p38γ, a molecular target of ImP, is activated in patients with primary sclerosing cholangitis (PSC), a long-term progressive disease of the liver and gallbladder (Panzitt et al., J. Hepatol. 72, 1122-1131, 2020). PSC increases the risk of various cancers, including liver cancer, gallbladder cancer, colorectal cancer, and cholangiocarcinoma. For example, ImP has been shown to be elevated in patients with colorectal cancer (Gao et al., Gastroenterology 163:1024-1037, e1029, 2022). Increased ImP levels have also been shown to be associated with the Göttingen minipig model of NASH (Lutzhoft et al., BMC Microbiol. 22, 287, 2020). These studies suggest that reducing ImP levels is a therapeutic target in the treatment of various gastrointestinal disorders and related conditions. [Overview of the project]

[0007] ImP is a known microbial histidine degradation product. Figure 1 shows the histidine degradation pathway in which urocanic acid reductase is involved in the conversion of urocanic acid to ImP.

[0008] By the method of the present invention, the inventors have identified compounds that are urocanate reductase inhibitors. Such compounds can inhibit the conversion of urocanate to ImP by urocanate reductase and are proposed herein for use in preventing, inhibiting, or treating disorders and conditions mediated by microbial urocanate reductase.

[0009] Therefore, a common objective is to provide compounds that can inhibit urocanate reductase, particularly bacterial urocanate reductase, and thus reduce ImP levels in the intestines, circulation, or body tissues.

[0010] Another general objective is to provide compounds useful for the prevention, inhibition, or treatment of disorders and conditions mediated by urocanate reductase, more specifically, disorders and conditions arising from and / or related to urocanate reductase-mediated ImP production. Such disorders and conditions include, but are not limited to, metabolic disorders, cardiovascular disorders, and gastrointestinal disorders. The use of such compounds in the treatment of patients suffering from such disorders and conditions is also an objective.

[0011] These and other objectives are satisfied by the embodiments disclosed herein.

[0012] In one embodiment, the present invention provides a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts for use in preventing, inhibiting, or treating conditions or disorders mediated by urocanate reductase, [ka] During the ceremony, A is a five-membered heteroaromatic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, wherein at least one of the heteroatoms is nitrogen. R 1 and R 2 but, -H; C 1-6 alkyl, preferably C 1-3 alkyl, such as -CH3, C 1-6 haloalkyl, preferably C 1-3 haloalkyl, such as -CF3, C 1-6 alkoxy, preferably C 1-3 alkoxy, such as -OCH3, halogen, preferably F, Cl, Br, or I, and -CN, are independently selected from, when present, each R 3 is, C 1-6 alkyl, preferably C 1-3 alkyl, such as -CH3, C 1-6 haloalkyl, preferably C 1-3 haloalkyl, such as -CF3, C 1-6 alkoxy, preferably C 1-3 alkoxy, such as -OCH3, halogen, preferably F, Cl, Br, or I, -OH, -CN, -NO2, -NR 5 R 6 (wherein R 5 and R 6 are independently selected from H and C 1-3 alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 (wherein R 7 , R 8 , and R 9 each is independently selected from H and C 1-3 alkyl, preferably selected from H and -CH3), are independently selected from, R 4 is, -OH, -SH, C 1-6Alkoxy, preferably C 1-3 Alkoxy, and -NR 10 R 11 (In the formula, R 10 and R 11 However, H and C 1-3 Selected independently from alkyl, preferably selected from H and -CH3, and n is an integer between 0 and 2.

[0013] In another embodiment, the present invention provides compounds of formula (I), stereoisomers, tautomers, and pharmaceutically acceptable salts thereof for use as pharmaceuticals.

[0014] In a further embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (I), its tautomers, stereoisomers, or pharmaceutically acceptable salts together with one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0015] In another embodiment, the present invention relates to novel compounds of formulas (II), (III), (IV), and (V) as described herein, their stereoisomers, tautomers, and pharmaceutically acceptable salts.

[0016] In a further embodiment, the present invention relates to compounds of formula (II), (III), (IV), or (V), or their stereoisomers, tautomers, or pharmaceutically acceptable salts, for use as pharmaceuticals.

[0017] In a further embodiment, the present invention relates to compounds of formula (II), (III), (IV), or (V), or their stereoisomers, tautomers, or pharmaceutically acceptable salts, for use in the prevention, inhibition, or treatment of urocanate reductase-mediated conditions or disorders.

[0018] In a further embodiment, the present invention relates to a method for preparing compounds of formula (II), (III), (IV), or (V), or stereoisomers, tautomers, or pharmaceutically acceptable salts thereof.

[0019] In a further embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (II), (III), (IV), or (V), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0020] The use of any of the compounds described herein, or their stereoisomers, tautomers, or pharmaceutically acceptable salts, in the manufacture of a pharmaceutical product for use in preventing, inhibiting, or treating a condition or disorder mediated by urocanic acid reductase constitutes a further aspect of the present invention.

[0021] A further aspect of the present invention is a method for preventing, inhibiting, or treating a condition or disorder mediated by urocanate reductase, comprising the step of administering to a patient (e.g., a human subject) in need of such treatment a pharmaceutically effective amount of any compound described herein or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof. [Modes for carrying out the invention]

[0022] definition As used herein, the term "alkyl" refers to a monovalent, saturated, linear, or branched carbon chain. Alkyl groups preferably contain 1 to 3 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, and isopropyl.

[0023] As used herein, the term "alkoxy" refers to an -O-alkyl group, where alkyl is as defined herein. The alkoxy group preferably contains 1 to 3 carbon atoms. Examples of alkoxy groups include methoxy, ethoxy, n-propoxy, and isopropoxy.

[0024] As used herein, the term "halogen" refers to F, Cl, Br, or I.

[0025] The term "haloalkyl" refers to an alkyl group as defined herein, in which at least one of the hydrogen atoms of the alkyl group is replaced by a halogen atom, preferably F, Cl, or Br. Examples of such groups include -CH2F, -CHF2, -CF3, -CCl3, -CHCl2, and -CH2CF3.

[0026] The term "heteroaromatic ring" refers to a heterocyclic aromatic group. A heterocyclic aromatic group comprises at least one heteroaromatic ring system. It contains at least one heteroatom selected from nitrogen, oxygen, and sulfur, and contains enough conjugated bonds to form the aromatic system. In the compounds described herein, a heteroaromatic ring is a five-membered ring containing 1 to 4 heteroatoms, at least one of which is nitrogen. Examples of such heteroaromatic rings include, but are not limited to, thiazolyl, pyrrolyl, triazolyl, imidazolyl, oxadiazolyl, pyrazolyl, tetrazolyl, and thiadiazolyl. Unless otherwise specified, any heteroaromatic ring referred to herein may be optionally substituted with one or more groups. If two or more substituents are present, they may be the same or different.

[0027] Unless otherwise specified, all substituents are independent of each other.

[0028] If the subscript is the integer 0 (i.e., zero), it is intended that the base pointed to by the subscript does not exist.

[0029] Unless otherwise specified, throughout this application and the attached claims, any given chemical formula or chemical name shall encompass all of its tautomers and stereoisomers, as well as racemates and mixtures of any of these forms in different proportions.

[0030] The compounds of the present invention may exist in the form of different stereoisomers. The term "stereoisomer" refers to a compound that has the same chemical composition but differs in terms of the spatial arrangement of atoms or groups. For example, a compound may contain one or more stereocenters and therefore may exist as different enantiomers or diastereoisomers. The term "enantiomer" refers to two stereoisomers of a compound that are mirror images of each other and cannot be superimposed. The term "diastereoisomer" refers to a stereoisomer having two or more stereocenters that are not mirror images of each other. The present invention is thought to extend to diastereoisomers and enantiomers, as well as racemic mixtures and enantiomer-rich mixtures in which the ratio of enantiomers is other than 1:1.

[0031] The compounds described herein can be decomposed into their enantiomers and / or diastereoisomers. For example, if they contain only one chiral center, they can be provided in the form of a racemic compound or a racemic mixture (a 50:50 mixture of enantiomers), or as pure enantiomers, i.e., in R or S form. Any of the compounds resulting as a racemic compound can be separated into their enantiomers by methods known in the art, such as column separation on a chiral phase, or by recrystallization from an optically active solvent. These compounds having at least two chiral carbon atoms can be decomposed into their diastereoisomers based on their physicochemical differences using methods known in themselves, for example, by chromatography and / or fractionation crystallization, and if these compounds are obtained in racemic form, they can then be decomposed into their enantiomers.

[0032] Because carbon-carbon double bonds are present in the compounds described herein, their isomers also include cis / trans or E / Z isomers. These may also be referred to as geometric isomers. In preferred embodiments, the compounds described herein include ring A and group -C(O)R 4 These are provided as specific geometric isomers that are in trans positions relative to each other.

[0033] As used herein, the term “tautomer” refers to structural isomers that readily interconvert. This may be due to chemical reactions involving, for example, the transfer of protons between single bonds and adjacent double bonds. In particular, this includes keto-enol tautomers and amide-imide acid tautomers, as well as tautomer forms of five-membered heterocyclic compounds containing two or more ring nitrogen atoms (e.g., imidazole, pyrazole, tetrazole, etc.). Depending on the conditions, a compound may exist primarily in one of its tautomers. Neutral and amphoteric forms of a compound are also encompassed by the term “tautomer” as used herein. These may arise, for example, from the transfer of protons from a carboxylic acid to an amine group within the molecule. The predominance of the amphoteric form of a compound depends on the pH. The present invention is thought to extend to the use of any tautomer and mixtures of tautomers of the compounds described herein.

[0034] As used herein, the term “pharmaceutically acceptable salt” refers to any pharmaceutically acceptable organic or inorganic salt of any of the compounds described herein. A pharmaceutically acceptable salt may contain one or more additional molecules, such as a counterion. The counterion may be any organic or inorganic group that stabilizes the charge of the parent compound. If the compound of the present invention is a base, a suitable pharmaceutically acceptable salt may be prepared by the reaction of a free base with an organic or inorganic acid. If the compound of the present invention is an acid, a suitable pharmaceutically acceptable salt may be prepared by the reaction of a free acid with an organic or inorganic base. Non-limiting examples of suitable salts are described herein.

[0035] The term "pharmaceutically acceptable" means that a compound or composition is chemically and / or toxicologically compatible with other components of the formulation or with the patient being treated (e.g., a human).

[0036] "Pharmaceutical composition" means any form of composition suitable for use for medical purposes.

[0037] As used herein, “treatment” includes any therapeutic use that may benefit humans or non-human animals (e.g., non-human mammals). While therapies of both humans and animals are within the scope of this invention, the invention is primarily intended for the treatment of humans. The treatment may relate to an existing disease or condition, or it may be preventative.

[0038] As used herein, “prevention” means any preventive treatment of a defined disorder in the subject. Specifically, this includes, in particular, preventing the occurrence of a disorder or reducing the risk of a disorder occurring in a subject who is susceptible to a disorder but has not yet been diagnosed with it.

[0039] As used herein, “inhibition” of a disability as defined in the subject includes (i) stopping the further occurrence (progression) of the disability or delaying its onset, and (ii) mitigating the disability, in particular causing regression of the disability until a desired outcome is reached.

[0040] As used herein, “pharmaceutically effective amount” refers to the amount that leads to the desired pharmacological and / or therapeutic effect, i.e., the amount of the agent that is effective in achieving its intended purpose. While the needs of individual patients may vary, determining the optimal range for an effective amount of the active agent is within the scope of the skills of those skilled in the art. In general, a dosage regimen for treating a disease or condition with any of the compounds described herein is selected according to a variety of factors, including the nature and severity of the medical condition.

[0041] As used herein, "urocanate reductase" refers to an enzyme that catalyzes the conversion of urocanate to ImP in the microbial histidine degradation pathway. In particular, it refers to an enzyme having classification EC 1.3.99.33. Urocanate reductase is an enzyme that is concentrated in the gut microbiota of subjects suffering from T2D or prediabetes. In these bacteria, urocanate reductase encoded by the gene urdA is involved in the conversion of urocanate to ImP (see Figure 4). Since no urdA homolog with more than 30% sequence identity has been identified in the human genome, the enzyme urocanate reductase does not exist in humans to the best of our knowledge. ImP-producing bacteria include, but are not limited to, Aerococcus urinae, Anaerococcus prevotii, Brevibacillus laterosporus, Eggerthella lenta, and Shewanella oneidensis.

[0042] References to “urocanate reductase activity” in this specification refer to the conversion of urocanate to ImP, or other enzymatic activities of urocanate reductase, or fragments thereof. References to “urocanate reductase inhibitors” or “inhibition of urocanate reductase” should be interpreted accordingly. Thus, a “urocanate reductase inhibitor” is a compound that reduces the conversion of urocanate to ImP or otherwise reduces the enzymatic activity of urocanate reductase. Such reductions do not need to be complete, but are typically at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80%, or at least 90% or at least 95%.

[0043] As used herein, “prediabetes” refers to either impaired fasting glycemia (IFG) or impaired glucose tolerance (IGT).

[0044] As used herein, “fasting blood glucose abnormalities” or “IFG” is a type of prediabetes in which a subject’s fasting blood glucose level is consistently above the normal range but below the diagnostic cutoff for formal diagnosis of type 2 diabetes. According to World Health Organization (WHO) standards, IFG is defined as a fasting plasma glucose level of 110–125 mg / dl, or 6.1–6.8 mmol / l.

[0045] As used herein, “impaired glucose tolerance” or “IGT” is a prediabetic state characterized by hyperglycemia associated with insulin resistance and an increased risk of cardiovascular disease. Like IFG, IGT can precede type 2 diabetes (T2D) by many years. According to the World Health Organization (WHO) and American Diabetes Association (AAD) criteria, IGT is defined as a 2-hour glucose level of 140–199 mg / dl, or 7.8–11.0 mmol / l, on a 75g oral glucose tolerance test. A patient is said to be in a state of IGT if their glucose level after 2 hours is moderately elevated but below the level required to qualify for T2D. Fasting glucose may be normal or moderately elevated, typically below 6.1 mmol / l.

[0046] As used herein, “Type 2 diabetes” or “T2D” is also known as type 2 diabetes mellitus and is a long-term metabolic disorder characterized by hyperglycemia, insulin resistance, and relative insulin deficiency. Long-term complications from hyperglycemia include heart disease, stroke, diabetic retinopathy which can lead to blindness, renal failure, and reduced blood flow to the limbs which can lead to amputation. The WHO definition of T2D is a symptomatic single elevated glucose reading, or two elevated readings, either fasting plasma glucose ≥ 7.0 mmol / l (126 mg / dl) or plasma glucose ≥ 11.1 mmol / l (200 mg / dl) in a glucose tolerance test 2 hours after oral administration. 1c The WHO definition of T2D for this condition is ≥48 mmol / mol. The table below summarizes the WHO diagnostic criteria for diabetes. [Table 1]

[0047] The inventors have found that the compounds described herein can act as inhibitors of urocanate reductase (e.g., urocanate reductase according to enzyme classification EC 1.3.99.33). This finding leads to the use of the compounds to treat or prevent conditions or diseases in subjects, e.g., humans, that are mediated by the activity of urocanate reductase. As inhibitors of the bacterial enzyme urocanate reductase, the compounds described herein are particularly suitable for inhibiting the production of harmful ImP and are useful compounds for preventing or treating disorders or conditions associated with elevated levels of ImP in the intestines, circulatory system, or body tissues, e.g., elevated levels of ImP in the blood (e.g., plasma or serum), urine, or feces. These include metabolic disorders (e.g., T2D, IGT, and prediabetes), cardiovascular disorders, and gastrointestinal disorders.

[0048] In one embodiment, the present invention relates to a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts for use in preventing, inhibiting, or treating conditions or disorders mediated by urocanate reductase. [ka] During the ceremony, A is a five-membered heteroaromatic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, wherein at least one of the heteroatoms is nitrogen. R 1 and R 2 but, -H; C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, for example -OCH3, Halogens, preferably F, Cl, Br, or I, and -CN, selected independently of, If present, each R 3 but, C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, for example -OCH3, Halogen, preferably F, Cl, Br, or I -OH, -CN, -NO2, -NR 5 R 6 (In the formula, R 5 and R 6 However, H and C 1-3 (Selected independently from alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 )(wherein, R 7, R 8 , and R 9 Each of them is H and C 1-3 Independently selected from alkyl, preferably selected from H and -CH3), independently selected from, R 4 but, -OH, -SH, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, and -NR 10 R 11 (In the formula, R 10 and R 11 However, H and C 1-3 Selected independently from alkyl, preferably selected from H and -CH3, and n is an integer between 0 and 2.

[0049] In formula (I), the heteroaromatic ring A is a base at any suitable point on the ring. [ka] It can be bonded to the ring nitrogen atom or to the carbon atoms in the ring. As understood, the bond may be via the ring nitrogen atom or to the carbon atoms in the ring. In one embodiment, heteroaromatic ring A is bonded to the ring via the carbon atoms in the ring. [ka] It is connected.

[0050] If present, any R 3 The group can be bonded to the heteroaromatic ring A at any suitable point on the ring. As understood, the bond may be via a ring nitrogen atom or via a carbon atom in the ring. In one embodiment, a single group R 3 A group exists (where n is 1) and is bonded to the heteroaromatic ring via a ring nitrogen atom. In another embodiment, a single group R 3 In yet another embodiment, two groups R 3There exist (where n is 2), and these are bonded to two carbon atoms in the heteroaromatic ring, for example, two adjacent carbon atoms.

[0051] In one embodiment, the compounds for use in the present invention are compounds of formula (I), stereoisomers thereof, tautomers thereof, and pharmaceutically acceptable salts, wherein A is a five-membered heteroaromatic ring containing 1 to 3 heteroatoms selected from nitrogen and sulfur. In one embodiment, A is a five-membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen and sulfur.

[0052] In one embodiment, the compounds for use in the present invention are compounds of formula (I), stereoisomers thereof, tautomers thereof, and pharmaceutically acceptable salts, wherein A is a five-membered heteroaromatic ring containing 1 to 3 nitrogen atoms. In one embodiment, A is a five-membered heteroaromatic ring containing 1 or 2 nitrogen atoms.

[0053] In one embodiment, the compounds for use in the present invention are the compound of formula (I), its stereoisomers, tautomers, and pharmaceutically acceptable salts, where A is optionally substituted with an imidazolyl, pyrazolyl, pyrrolyl, or thiazolyl group. In one embodiment, A is optionally substituted with an imidazolyl, pyrazolyl, or pyrrolyl group. In another embodiment, A is optionally substituted with 4-imidazolyl, 3-pyrazolyl, 4-pyrazolyl, 3-pyrolyl, or 4-thiazolyl group.

[0054] As can be understood, any optional substituent present in ring A is the group R as defined herein. 3 In one embodiment, each R 3 C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 A component is independently selected from alkoxy, F, Cl, Br, I, -OH, -NH2, and -C(=NH)(NH2). In one embodiment, each R 3is independently selected from -CH3, -CF3, -OCH3, -OH, -NH2, and -C(=NH)(NH2). In one embodiment, each R 3 is independently selected from -CH3 and -NH2.

[0055] The substituent R 3 When it is C 1-6 alkyl, it may be attached to either the ring nitrogen atom or the ring carbon in group A. In one embodiment, any substituent R 3 present is attached to a carbon atom forming part of the ring.

[0056] The number of substituents R 3 present on ring A can vary. In one embodiment, there are no substituents on ring A (i.e., n is 0). In one embodiment, a single substituent is present, i.e., n is 1.

[0057] In one embodiment, the compound for use in the present invention is a compound of formula (I), its stereoisomers, tautomers and pharmaceutically acceptable salts, wherein n is other than 0 and R 3 is C 1-3 alkyl (e.g., -CH3 or -CH2CH3), C 1-3 haloalkyl (e.g., -CF3), C 1-3 alkoxy (e.g., -OCH3), F, Cl, -NH2, and -OH.

[0058] In one embodiment, the compound for use in the present invention is a compound of formula (I), its stereoisomers, tautomers, and pharmaceutically acceptable salts, wherein n is 1 and R 3 is C 1-3 alkyl (e.g., -CH3 or -CH2CH3), C 1-3 haloalkyl (e.g., -CF3), C 1-3 alkoxy (e.g., -OCH3), F, Cl, -NH2, and -OH, preferably R 3 is either -CH3 or -NH2.

[0059] In one embodiment, the compound for use in the present invention is a compound of formula (I), its stereoisomers, tautomers, and pharmaceutically acceptable salts, wherein R 1 and R 2 are independently selected from -H, C 1-3 alkyl (e.g., -CH3 or -CH2CH3), C 1-3 haloalkyl (e.g., -CF3), F, and Cl.

[0060] In one embodiment, R 1 is H, -CH3, -CH2CH3, or F.

[0061] In one embodiment, R 1 is H.

[0062] In one embodiment, R 2 is H, -CH3, -CH2CH3, or F.

[0063] In one embodiment, R 2 is H, -CH3, or -CH2CH3.

[0064] In one embodiment, R 2 is -CH3 or -CH2CH3, preferably -CH3.

[0065] In one embodiment, R 2 is other than H.

[0066] In one embodiment, the compound for use in the present invention is a compound of formula (I), its stereoisomers, tautomers, and pharmaceutically acceptable salts, wherein R 4 is -OH or C 1-3 alkoxy. In one embodiment, R 4 is OH.

[0067] In one embodiment, the compound for use in the present invention is a heteroaromatic ring A and a group -C(O)R 4It is one of the compounds of formula (I) described herein, wherein the two elements are trans relative to each other.

[0068] In one embodiment, the compounds used in the present invention are compounds of formula (II), their stereoisomers, tautomers, and pharmaceutically acceptable salts. [ka] In the formula, R 1 ~R 2 , R 3 , and R 4 However, as defined herein, and m is an integer between 0 and 2, preferably 0 or 1.

[0069] In one embodiment, the compound used in the present invention is an imidazolyl ring and the group -C(O)R 4 The compound of formula (II) described herein is trans relative to each other.

[0070] In one embodiment, the compounds used in the present invention are compounds of formula (III), their stereoisomers, tautomers, and pharmaceutically acceptable salts. [ka] In the formula, R 1 ~R 2 , R 3 , and R 4 However, as defined herein, and p is an integer between 0 and 2, preferably 0 or 1.

[0071] In one embodiment, the compound used in the present invention is a pyrazolyl ring and the group -C(O)R 4 However, these are compounds of formula (III) as described herein, which are trans relative to each other.

[0072] In one embodiment, the compounds used in the present invention are the compound of formula (IV), its stereoisomers, tautomers, and pharmaceutically acceptable salts. [ka] In the formula, R 1 ~R 2 , R 3 , and R 4 However, as defined herein, and q is an integer between 0 and 2, preferably 0 or 1.

[0073] In one embodiment, the compound used in the present invention is a pyrrolyl ring and the group -C(O)R 4 However, the compounds are trans compounds of formula (IV) as described herein.

[0074] In one embodiment, the compounds used in the present invention are the compound of formula (V), its stereoisomers, tautomers, and pharmaceutically acceptable salts thereof. [ka] In the formula, R 1 ~R 2 , R 3 , and R 4 However, as defined herein, and s is an integer between 0 and 2, preferably 0 or 1.

[0075] In one embodiment, the compound used in the present invention is a thiazolyl ring and a group -C(O)R 4 However, these are compounds of formula (V) described herein, which are trans relative to each other.

[0076] Examples of compounds for use according to the present invention include, but are not limited to, the following stereoisomers, tautomers, and pharmaceutically acceptable salts thereof. [Table 2-1] [Table 2-2] [Table 2-3]

[0077] Preferred for use in the present invention are specific geometric isomers of compounds 1 to 28 shown in the table above, their tautomers, and pharmaceutically acceptable salts thereof, in which the five-membered heteroaromatic ring is trans to the -C(O)OH group.

[0078] Preferred for use in the present invention are compound numbers 1 to 12, their stereoisomers, tautomers, and pharmaceutically acceptable salts. Particularly preferred for use in the present invention are specific geometric isomers of compound numbers 1 to 12 shown in the table above, in which the five-membered heteroaromatic ring is trans to the -C(O)OH group, and pharmaceutically acceptable salts thereof.

[0079] The specific compounds described herein are novel and form further aspects of the present invention. Accordingly, in a further embodiment, the present invention provides specific novel compounds of formulas (II), (IIa), (III), (IIIa), (IV), (IVa), (V), and (Va), their stereoisomers, tautomers, and pharmaceutically acceptable salts thereof.

[0080] The novel compound of formula (II) is a compound having the following structural formula: [ka] During the ceremony, R 1 and R 2 but, -H; C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, for example -OCH3, Halogens, preferably F, Cl, Br, or I, and -CN, selected independently of, If present, each R 3 but, C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, for example -OCH3, Halogen, preferably F, Cl, Br, or I -OH, -CN, -NO2, -NR 5 R 6 (In the formula, R 5 and R 6 However, H and C 1-3 (Selected independently from alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 )(wherein, R 7 , R 8 , and R 9 Each of them is H and C 1-3 Independently selected from alkyl, preferably selected from H and -CH3), independently selected from, R 4 but, -OH, -SH, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, and -NR 10 R 11 (In the formula, R 10 and R 11 However, H and C 1-3 Selected independently from alkyl, preferably selected from H and -CH3, and m is an integer between 0 and 2, preferably 0 or 1. However, if the compound is [ka] That's surprising.

[0081] In one embodiment, the novel compound of formula (II) is either m = 0 (i.e., the heteroaromatic ring is unsubstituted) or m = 1, R 3 is base-NR 5 R 6 The compound is preferably -NH2.

[0082] In one embodiment, the novel compound of formula (II) is R 1 and R 2 However, independently, -H and C 1-6 Selected from alkyl groups, preferably -H and C 1-3 The compound is selected from alkyl groups, for example, from -H and -CH3.

[0083] In one embodiment, the novel compound of formula (II) is R 4 It is a compound in which OH is present.

[0084] In one embodiment, the novel compound of formula (II) comprises an imidazolyl ring and the group -C(O)R 4 However, they are compounds that are trans relative to each other.

[0085] The novel compound of formula (IIa) is a compound having the following structural formula, its stereoisomer, tautomer, or pharmaceutically acceptable salt, [ka] During the ceremony, If present, each R 3 but, C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C1-6 Alkoxy, preferably C 1-3 Alkoxy, such as -OCH3, Halogen, preferably F, Cl, Br, or I, -OH, -CN, -NO2, -NR 5 R 6 (wherein R 5 and R 6 are independently selected from H and C 1-3 alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 (wherein R 7 , R 8 , and R 9 each is independently selected from H and C 1-3 alkyl, preferably selected from H and -CH3), and m is an integer from 0 to 2, preferably 0 or 1.

[0086] In one embodiment, the novel compound of formula (IIa) is such that m is 0, or m is 1, and R 3 is the group -NR 5 R 6 , preferably -NH2, or R 3 is C 1-3 alkyl, preferably -CH3.

[0087] In one embodiment, the novel compound of formula (IIa) is a compound in which the imidazolyl ring and the group -C(O)OH are trans to each other.

[0088] The compound of formula (III) is a compound having the following structural formula,

Chemical formula

[0089] In one embodiment, the novel compound of formula (III) has p = 0 (i.e., the heteroaromatic ring is unsubstituted), or p = 1, R 3 C 1-6 Alkyl, preferably C 1-3 It is an alkyl compound, for example, a -CH3 compound.

[0090] In one embodiment, the novel compound of formula (III) is R 1 and R 2 However, -H and C 1-6 From alkyl, preferably -H and C 1-3 The compound is independently selected from alkyl groups, for example, -H, -CH3, and -CH2CH3.

[0091] In one embodiment, the novel compound of formula (III) is R 4 It is a compound in which OH is present.

[0092] In one embodiment, the novel compound of formula (III) comprises a pyrazolyl ring and the group -C(O)R 4 These are compounds in which the two elements are trans relative to each other.

[0093] The novel compound of formula (IIIa) is a compound having the following structural formula, its stereoisomer, tautomer, or pharmaceutically acceptable salt, [ka] In the formula, when present, each R 3 is C 1-6 alkyl, preferably C 1-3 alkyl, for example -CH3, C 1-6 haloalkyl, preferably C 1-3 haloalkyl, for example -CF3, C 1-6 alkoxy, preferably C 1-3 alkoxy, for example -OCH3, halogen, preferably F, Cl, Br, or I, -OH, -CN, -NO2, -NR 5 R 6 (wherein R 5 and R 6 are independently selected from H and C 1-3 alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 (wherein R 7 , R 8 , and R 9 each is independently selected from H and C 1-3 alkyl, preferably selected from H and -CH3), are independently selected from, and p is an integer from 0 to 2, preferably 0 or 1.

[0094] In one embodiment, the novel compound of formula (IIIa) is a compound wherein p is 0, or p is 1 and R 3 is C 1-3 alkyl, preferably -CH3.

[0095] [[ID={74]]In one embodiment, the novel compound of formula (IIIa) is a compound wherein the pyrazolyl ring and the group -C(O)OH are trans to each other.

[0096] The compound of formula (IV) is a compound having the following structural formula, [ka] During the ceremony, R 1 and R 2 but, -H; C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, e.g., -OCH3, Halogens, preferably F, Cl, Br, or I, and -CN, selected independently of, If present, each R 3 but, C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, e.g., -OCH3, Halogen, preferably F, Cl, Br, or I -OH, -CN, -NO2, -NR 5 R 6 (In the formula, R 5 and R 6 However, H and C 1-3 (Selected independently from alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 )(wherein, R 7 , R 8 , and R 9Each of them is H and C 1-3 Independently selected from alkyl, preferably selected from H and -CH3), independently selected from, R 4 but, -OH, -SH, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, and -NR 10 R 11 (In the formula, R 10 and R 11 However, H and C 1-3 Selected independently from alkyl, preferably selected from H and -CH3, and q is an integer between 0 and 2, preferably 0 or 1. However, the compound of formula (IV) [ka] That's surprising.

[0097] In one embodiment, the novel compound of formula (IV) is a compound in which q is 0 (i.e., the heteroaromatic ring is unsubstituted).

[0098] In one embodiment, the novel compound of formula (IV) is R 1 and R 2 However, -H and C 1-6 From alkyl, preferably -H and C 1-3 The compound is independently selected from alkyl groups, for example, from -H and -CH3.

[0099] In one embodiment, the novel compound of formula (IV) is R 4 It is a compound in which OH is present.

[0100] In one embodiment, the novel compound of formula (IV) comprises a pyrrolyl ring and the group -C(O)R 4 These are compounds in which the two elements are trans relative to each other.

[0101] The novel compound of formula (IVa) is a compound having the following structural formula, its stereoisomer, tautomer, or pharmaceutically acceptable salt, [ka] During the ceremony, Each R 3 but, C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, e.g., -OCH3, Halogen, preferably F, Cl, Br, or I -OH, -CN, -NO2, -NR 5 R 6 (In the formula, R 5 and R 6 However, H and C 1-3 (Selected independently from alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 )(wherein, R 7 , R 8 , and R 9 Each of them is H and C 1-3 Independently selected from alkyl, preferably selected from H and -CH3), and q is 1 or 2, However, the compound of formula (IVa) [ka] That's surprising.

[0102] In one embodiment, the novel compound of formula (IVa) is a compound in which the pyrrolyl ring and the group -C(O)OH are trans relative to each other.

[0103] The compound of formula (V) is a compound having the following structural formula: [ka] During the ceremony, R 1 and R 2 but, -H; C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, e.g., -OCH3, Halogens, preferably F, Cl, Br, or I, and -CN, selected independently of, If present, each R 3 but, C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, e.g., -OCH3, Halogen, preferably F, Cl, Br, or I -OH, -CN, -NO2, -NR 5 R 6 (In the formula, R 5 and R 6 However, H and C 1-3 (Selected independently from alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 )(wherein, R 7 , R8 , and R 9 Each of them is H and C 1-3 Independently selected from alkyl, preferably selected from H and -CH3), independently selected from, R 4 but, -OH, -SH, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, and -NR 10 R 11 (In the formula, R 10 and R 11 However, H and C 1-3 Selected independently from alkyl, preferably selected from H and -CH3, and s is an integer between 0 and 2, preferably 0 or 1. However, if the compound is [ka] That's surprising.

[0104] In one embodiment, the novel compound of formula (V) has s = 1, and R 3 is base-NR 5 R 6 The compound is preferably -NH2.

[0105] In one embodiment, the novel compound of formula (V) is R 1 and R 2 It is a compound in which both atoms are -H.

[0106] In one embodiment, the novel compound of formula (V) is R 4 It is a compound in which OH is present.

[0107] In one embodiment, the novel compound of formula (V) comprises a thiazolyl ring and the group -C(O)R 4 However, they are compounds that are trans relative to each other.

[0108] The novel compound of formula (Va) is a compound having the following structural formula, its stereoisomer, tautomer, or pharmaceutically acceptable salt, [ka] During the ceremony, If present, each R 3 but, C 1-6 Alkyl, preferably C 1-3 Alkyl, for example -CH3, C 1-6 Haloalkyl, preferably C 1-3 Haloalkyl, e.g., -CF3, C 1-6 Alkoxy, preferably C 1-3 Alkoxy, e.g., -OCH3, Halogen, preferably F, Cl, Br, or I -OH, -CN, -NO2, -NR 5 R 6 (In the formula, R 5 and R 6 However, H and C 1-3 (Selected independently from alkyl, preferably selected from H and -CH3), and -C(=NR 7 )(NR 8 R 9 )(wherein, R 7 , R 8 , and R 9 Each of them is H and C 1-3 Independently selected from alkyl, preferably selected from H and -CH3), and s is an integer between 0 and 2, preferably 0 or 1. However, the compound of formula (Va) [ka] That's surprising.

[0109] In one embodiment, the novel compound of formula (Va) is a compound in which the thiazolyl ring and the group -C(O)OH are trans relative to each other.

[0110] In one embodiment, the compounds of formula (II) according to the present invention include the following compounds, their stereoisomers, tautomers, and pharmaceutically acceptable salts. [Table 3]

[0111] In one embodiment, the compounds of formula (II) according to the present invention are compounds selected from compound numbers 5, 8, 15-21, 25, and 28 in which the imidazolyl ring and the -C(O)OH group are trans to each other, their tautomers, and pharmaceutically acceptable salts thereof.

[0112] In one embodiment, the compounds of formula (III) according to the present invention include the following compounds, their stereoisomers, tautomers, and pharmaceutically acceptable salts: [Table 4]

[0113] In one embodiment, the compound of formula (III) according to the present invention is a compound selected from compound numbers 1, 4, 6, 26, and 27 in which the pyrazolyl ring and the -C(O)OH group are trans relative to each other, their tautomers, and pharmaceutically acceptable salts thereof.

[0114] In one embodiment, the compounds of formula (IV) according to the present invention include the following compounds, their stereoisomers, tautomers, and pharmaceutically acceptable salts: [Table 5]

[0115] In one embodiment, the compound of formula (IV) according to the present invention is a compound selected from compound numbers 22 and 23 in which the pyrrolyl ring and the -C(O)OH group are trans relative to each other, their tautomers, and their pharmaceutically acceptable salts.

[0116] In one embodiment, the compounds of formula (V) according to the present invention include the following compounds, their stereoisomers, tautomers, and pharmaceutically acceptable salts. [Table 6]

[0117] In one embodiment, the compound of formula (V) according to the present invention is compound number 2 and its pharmaceutically acceptable salts, in which the thiazolyl ring and the -C(O)OH group are trans relative to each other.

[0118] Novel compounds of formulas (II), (IIa), (III), (IIIa), (IV), (IVa), (V), and (Va) according to the present invention, their stereoisomers, and their tautomers may further be provided in the form of pharmaceutically acceptable salts.

[0119] In a further embodiment, the present invention provides novel compounds of formulas (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) as defined in the specification, or stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, for use as pharmaceuticals.

[0120] In a further embodiment, the present invention provides novel compounds of formulas (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) as defined herein, or stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, for use in preventing, inhibiting, or treating conditions or disorders mediated by urocanate reductase.

[0121] In a further embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) as defined in the specification, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0122] The use of novel compounds of formulas (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) as defined herein, or their stereoisomers, tautomers, or pharmaceutically acceptable salts, in the manufacture of pharmaceuticals for use in preventing, inhibiting, or treating conditions or disorders mediated by urocanate reductase, forms a further aspect of the present invention.

[0123] A method for preventing, inhibiting or treating a condition or disorder mediated by urocanate reductase, comprising the step of administering to a patient (e.g., a human subject) in need of such treatment a pharmaceutically effective amount of a novel compound of formula (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) as defined herein, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, forms a further aspect of the present invention.

[0124] Any of the compounds described herein can be converted to salts thereof, in particular pharmaceutically acceptable salts thereof having an inorganic or organic acid or base.

[0125] Suitable pharmaceutically acceptable salts of the compounds described herein are, for example, acid addition salts of the compounds that are sufficiently basic, such as acid addition salts with inorganic or organic acids. Acids that can be used for this purpose may include hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, sulfonic acid, methanesulfonic acid, phosphoric acid, fumaric acid, succinic acid, lactic acid, citric acid, tartaric acid, malic acid, malonic acid, maleic acid, acetic acid, trifluoroacetic acid and ascorbic acid, p-toluenesulfonic acid, 2-mesitylenesulfonic acid, 1,2-ethanedisulfonic acid, adipic acid, aspartic acid, benzenesulfonic acid, benzoic acid, ethanesulfonic acid, or nicotinic acid.

[0126] Furthermore, suitable pharmaceutically acceptable salts of the compounds described herein include, for example, base addition salts of the compounds that are sufficiently acidic, such as metal salts such as sodium, potassium, calcium, magnesium, and aluminum salts, ammonium salts, or salts with organic bases that provide physiologically acceptable cations, such as quaternary ammonium hydroxides, such as methylamine, ethylamine, diethylamine, trimethylamine, tert-butylamine, trimethylamine, dibenzylamine, N,N-dibenzylethylamine, cyclohexylethylamine, and tris-(2-hydroxyethyl )amines, hydroxyethyldiethylamine, (1R,2S)-2-hydroxyinden-1-amine, morpholine, N-methylpiperidine, N-ethylpiperidine, piperazine, methylpiperazine, adamantylamine, choline hydroxide, tetrabutylammonium hydroxide, tris-(hydroxymethyl)methylamine hydroxide, L-arginine, N-methyl D-glucamine, lysine or arginine, and organic amines, such as diethylamine, trimethylamine, ethanolamine, diethanolamine, cyclohexylamine, and dicyclohexylamine.

[0127] The procedure for salt formation is conventional in the field.

[0128] Naturally, the compounds described herein may exist in various stereoisomeric forms, including enantiomers, diastereoisomers, and mixtures thereof. The present invention encompasses all optical isomers and mixtures of optical isomers of the compounds described herein. Therefore, compounds existing as diastereoisomers, racemic compounds, and / or enantiomers are within the scope of the present invention. In preferred embodiments, the compounds described herein include a heteroaromatic ring A and the group -C(O)R 4 These are provided as specific geometric isomers that are trans relative to each other.

[0129] Compounds of formula (I) are known in the art or can be prepared by methods known to those skilled in the art. Many of these compounds are commercially available from sources including Merck Sigma-Aldrich (St. Louis, USA), Enamine LLC (Cincinnati, USA), Chemspace-FCH Group (Riga, Latvia), Auroro Fine Chemicals LLC (San Diego, USA), and Rare Chemicals GmbH (Kiel, Germany).

[0130] Any of the compounds described herein that are not known in the art, including the compounds of formulas (II), (IIa), (III), (IIIa), (IV), (IVa), (V), and (Va), can be prepared from readily available starting materials using synthetic methods known in the art, for example, methods described in well-known textbooks, such as Advanced Organic Chemistry (March, Wiley Interscience, 8th edition 2019) or Advanced Organic Chemistry (Carey and Sundberg, KA / PP, Part B, 5th edition 2007).

[0131] The following scheme illustrates a general method for preparing the compounds described herein. Such methods for preparing the compounds of formulas (II), (IIa), (III), (IIIa), (IV), (IVa), (V), and (Va) form further embodiments of the present invention. The compounds used as starting materials may be known from the literature or commercially available. Alternatively, they can be readily obtained by methods known from the literature. As can be understood, compounds can be prepared using different starting materials, different reagents, and / or different reaction conditions using other synthetic routes. A more detailed description of methods for preparing compounds according to the present invention can be found in the examples.

[0132] Scheme 1: [ka]

[0133] In Scheme 1, R 3 And m are as defined herein.

[0134] Scheme 2: [ka]

[0135] In Scheme 2, R 3 And m are as defined herein.

[0136] Scheme 3: [ka]

[0137] In scheme 3, R 3 and p are as defined herein.

[0138] Scheme 4: [ka]

[0139] In scheme 4, R 3 and p are as defined herein.

[0140] Scheme 5: [ka]

[0141] In scheme 5, R 3 And q are as defined herein.

[0142] Any method for preparing novel compounds of formula (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), which includes a step of deprotecting a protected derivative thereof, also forms part of the present invention.

[0143] The compounds described herein possess beneficial pharmacological properties, particularly inhibitory effects against microbial urocanate reductase. In terms of their ability to inhibit microbial urocanate reductase, the compounds described herein are suitable for the prevention, inhibition, or treatment of any condition or disease mediated by the activity of microbial urocanate reductase. As described herein, microbial urocanate reductase plays a central role in the microbial histidine degradation pathway, particularly in the production of ImP. Therefore, the compounds described herein are particularly suitable for the prevention, inhibition, or treatment of conditions involving microbial urocanate reductase, especially conditions arising from and / or related to the production of ImP. Such conditions include, but are not limited to, metabolic disorders, cardiovascular disorders, and gastrointestinal disorders.

[0144] Metabolic disorders may be associated with changes in the structure and function of the gut microbiota. The microbial metabolite ImP is present at higher concentrations in individuals with T2D and impaired glucose tolerance (IGT) compared to healthy individuals with normal glucose tolerance (NGT). Therefore, ImP contributes to the pathogenesis of T2D and IGT.

[0145] The compounds described herein are used for the prevention, inhibition, or treatment of the following metabolic disorders: type 2 diabetes (T2D), prediabetes, impaired glucose tolerance (IGT), and impaired fasting blood glucose (IFG).

[0146] Due to their activity in inhibiting urocanate reductase and subsequently reducing ImP, the compounds are also expected to be effective in treating complications associated with T2D, such as diabetic nephropathy, non-alcoholic fatty liver disease and steatohepatitis, diabetic retinopathy, and diabetic neuropathy. Furthermore, ImP is associated with poor glycemic control after metformin treatment in patients with T2D, and ImP inhibits the glucose-lowering effect of metformin in mice (Koh et al., Cell Metabolism 32(4):643-653, 2020). Therefore, blocking ImP production may improve the therapeutic effect of metformin. Accordingly, the compounds described herein may be used in combination therapy with metformin for the treatment of T2D.

[0147] The compounds described herein can also be used to prevent, inhibit, or treat gastrointestinal disorders associated with elevated ImP levels. Treatable gastrointestinal disorders include disorders of the digestive system, comprising the gastrointestinal tract (esophagus, stomach, small intestine, and large intestine) and digestive accessory organs, including the pancreas, gallbladder, and liver. Inflammatory gastrointestinal conditions treatable by the present invention include inflammatory bowel disease (IBD). As used herein, IBD refers to a group of inflammatory conditions of the colon and small intestine, including Crohn's disease and ulcerative colitis. Treatable hepatic or biliary conditions include primary sclerosing cholangitis (PSC). PSC is a long-term, progressive disease of the liver and gallbladder characterized by inflammation and scarring of the bile ducts (from which bile is normally discharged from the gallbladder). Bile duct scarring narrows the ducts, reducing the flow of bile into the intestines. This disruption of bile duct function can lead to cirrhosis and liver failure. PSC also increases the risk of various cancers, including liver cancer, gallbladder cancer, colorectal cancer, and bile duct cancer. Many patients with PSC also have IBD, typically ulcerative colitis.

[0148] Cardiovascular diseases and disorders that can be treated according to the present invention affect the heart itself or the vascular system, particularly the veins and arteries leading to the heart. Examples of such diseases and disorders include, but are not limited to, cardiovascular diseases, myocardial fibrosis, heart failure, and diabetic cardiomyopathy. Myocardial fibrosis refers to excessive deposition of extracellular matrix in the myocardium or abnormal thickening of the heart valves. This can cause the myocardium to harden, reduce its flexibility, and lead to heart failure.

[0149] Treatment, prevention, or inhibition of cardiovascular disease, particularly prevention of heart failure, constitutes a preferred embodiment of the present invention.

[0150] Accordingly, in further aspects, the present invention provides compounds for use in therapy as described herein. Unless otherwise specified, the term “therapy” as used herein is intended to include treatment, prevention, and inhibition.

[0151] In a further embodiment, the present invention provides compounds described herein for use in the prevention, inhibition, or treatment of any of the conditions described herein.

[0152] In another embodiment, the present invention provides the use of the compounds described herein in the manufacture of a pharmaceutical for use in a method of treating, preventing, or inhibiting any of the conditions described herein.

[0153] Also provided are methods for treating a human or non-human animal body to combat or prevent any of the conditions described herein, the methods comprising the step of administering an effective amount of a compound described herein to the body. The patient may be human.

[0154] The compounds described herein may be administered to subjects (e.g., patients) suffering from gastrointestinal disorders, cardiovascular diseases, or metabolic disorders, or to subjects (e.g., patients) at risk of developing such disorders.

[0155] In one embodiment, any use or method described herein may further include the step of determining whether a subject has or is at risk of developing one of the disorders described herein, based on a determined amount or concentration of ImP in a body sample from the subject, for example, in a body fluid sample. Examples of body fluid samples include blood samples, plasma samples, serum samples, urine samples, and fecal samples. In one embodiment, ImP may be used as a biomarker to identify individuals who may have T2D or IGT, or who are at risk of having these metabolic disorders. The amount of ImP in a body sample can be determined to identify such individuals, as described in International Publication No. 2018 / 097793, the entirety of which is incorporated herein by reference. The determined amount or concentration of ImP is compared to a threshold concentration, and if the concentration of ImP is above the threshold, the subject is determined to have or be at risk of having T2D or IGT. ImP measurement may also be performed to monitor and / or control the treatment of any subject according to any aspect of the present invention described herein.

[0156] The subjects treated by the embodiments of the present invention are mammalian subjects, preferably human subjects. However, the embodiments may also be applied to other mammalian subjects that may suffer from any of the conditions described herein, for example, metabolic disorders such as T2D or IGT. Therefore, the embodiments can also be used for veterinary purposes to treat animal subjects that are suffering from or at risk of suffering from any of the conditions or disorders described herein, such as T2D or IGT.

[0157] For use in therapeutic or prophylactic treatment, the compounds described herein may be administered alone or in combination with pharmaceutically acceptable carriers, excipients, or diluents. Typically, these are formulated as pharmaceutical formulations.

[0158] In a further embodiment, the present invention therefore provides a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, excipients, or diluents of a compound described herein, its stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

[0159] Acceptable carriers, excipients, and diluents for therapeutic use are well known in the art and can be selected in relation to the intended route of administration and standard pharmacopoeia. Examples include binders, lubricants, suspending agents, coatings, solubilizers, preservatives, wetting agents, emulsifiers, surfactants, sweeteners, colorants, flavoring agents, antioxidants, odorants, buffers, stabilizers, and / or salts.

[0160] The compounds for use in the present invention can be formulated with one or more conventional carriers and / or excipients according to techniques well known in the art. Typically, the compositions are adapted for enteral administration, e.g., oral or rectal administration.

[0161] For example, these can be formulated using conventional oral dosage forms such as tablets, coated tablets, pills, hard or soft capsules, powders (e.g., powders that can be reconstituted with water), granules, liquid formulations, aqueous or oily solutions, dispersions, aqueous or oily suspensions, syrups, elixirs, lozenges, and emulsions, with conventional excipients such as solvents, diluents, binders, sweeteners, fragrances, pH adjusters, viscosity modifiers, and antioxidants. Suitable excipients may include, for example, corn starch, lactose, glucose, microcrystalline cellulose, magnesium stearate, polyvinylpyrrolidone, citric acid, tartaric acid, water, ethanol, glycerol, sorbitol, polyethylene glycol, propylene glycol, cetyl stearyl alcohol, carboxymethylcellulose, or fatty substances such as saturated fats or suitable mixtures thereof. Depending on the dosage form, the product may be provided in the form of a pouch or vial containing the pharmaceutical composition. Formulations may be prepared using conventional techniques such as dissolution and / or mixing procedures and tableting.

[0162] Alternatively, the compound may be formulated in a conventional rectal administration form, such as a suppository. When provided in suppository form, the compound is formulated in a suppository base. Any known suppository base, including both water-soluble or water-miscible bases and fatty bases, can be used.

[0163] The use of orally administered compositions, such as tablets, coated tablets, capsules, and syrups, is particularly preferred. In one embodiment, such a dosage form may be formulated for delayed release, for example, for the release of the active compound in the colon. Enteric coatings that prevent the dissolution or disintegration of tablets or capsules in gastric juice are well known in the art, and any such coating may be used.

[0164] The dosage required to achieve the desired activity of the compounds described herein depends on various factors, including the compound selected, the method and frequency of administration, whether the treatment is therapeutic or prophylactic, and the nature and severity of the disease or condition. Typically, a physician determines the actual dosage best suited to the individual patient. Specific dose levels and frequencies of administration for a particular patient may vary and depend on factors such as the activity of the specific compound used, its metabolic stability and duration of action, the relationship between the compound's absorption and potency, the patient's age, the method and timing of administration, and the severity of the specific condition and any associated comorbidities (such as chronic kidney disease (CKD)). Compounds and / or pharmaceutical compositions may be administered according to regimens of once to ten times daily, such as once or twice daily. For oral and parenteral administration to human patients, the daily dose level of the drug may be a single dose or divided doses.

[0165] Typically, the dose may be administered once, twice, or three or more times daily. The compound may be administered to humans in doses ranging from 0.001 mg to 100 mg per kg of body weight per day, for example, in the range of 0.01 mg to 100 mg per kg of body weight per day, but this will inevitably vary depending on the weight, sex and condition of the person being treated, the disease condition being treated, and the specific route of administration selected. However, dose levels ranging from 0.1 mg to 10 mg per kg of body weight per day are most preferably used in humans, either as a single dose or in divided doses. For example, such doses may be appropriate for the treatment of metabolic disorders, cardiovascular metabolic disorders, and fibrotic diseases.

[0166] The pharmacological properties of the compounds of the present invention can be analyzed using standard assays for functional activity. Detailed protocols for testing the compounds of the present invention are provided in the examples. [Examples]

[0167] The present invention will now be described in more detail in the following non-limiting embodiments and with reference to the accompanying drawings. [Brief explanation of the drawing]

[0168] [Figure 1] Imidazole propionate as a histidine degradation product. The figure shows the histidine degradation pathway.

[0169] The chemical reactions described in the examples can be readily adapted to prepare other compounds for use in accordance with the present invention, for example, by using other reagents known in the art, by modifying the reaction conditions, and / or by selecting any suitable protecting group.

[0170] 1¹H NMR spectra were obtained using a Bruker-AVANACE-II (300 MHz) or -III (400 MHz) spectrometer with TopSpin software from Bruker BioSpin Corp, Germany. HPLC: Chromatograms were obtained using an Agilent 1200 series HPLC system, Agilent Technologies, USA. LC-MS: Chromatograms were recorded with an Agilent 1200 series with LC / MSD Trap XCT plus, USA.

[0171] Abbreviation: ACN: Acetonitrile BOC: Butyloxycarbonyl DCM: Dichloromethane DIPEA: N,N-diisopropylethylamine DMAP: 4-dimethylaminopyridine DMF: Dimethylformamide DMSO: Dimethyl sulfoxide EDC.HCl: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride HEPES / Tris:2-[4-(2-hydroxyethyl)piperazine-1-yl]ethanesulfonic acid / 2-amino-2-hydroxymethyl-propane-1,3-diol HOBt: Hydroxybenzotriazole HPLC: High-Pressure Liquid Chromatography KHMDS: Potassium bis(trimethylsilyl)amide LCMS: Liquid Chromatography Mass Spectrometry MTBE: Methyl tertiary butyl ether MR-1: The name of the Shewanella oneidensis strain used. NMR: Nuclear magnetic resonance MQ Pd(DPPF)Cl2-CH2-Cl2:[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex with dichloromethane SEM-Cl:2-(trimethylsilyl)ethoxymethylchloride TEA: Triethylamine TFA: Trifluoroacetic acid THF: Tetrahydrofuran TPGS-750 M: DL-α-Tocopherol Methoxypolyethylene Glycol Succinic Acid Solution TLC: Thin-layer chromatography

[0172] Example 1 Synthesis of -(E)-3-(4-pyrazolyl)-2-butenoic acid E (Compound 1) Step 1: Synthesis of 1-(1-{[2-(trimethylsilyl)ethoxy]methyl}-4-pyrazolyl)-1-ethanone (B): [ka]

[0173] To a stirred solution of (1H-pyrazole-4-yl)-ethanone (A) (250 mg, 2.27 mmol, 1 equivalent) in THF (5 mL), sodium hydride (60% in mineral oil) (100 mg, 2.5 mmol, 1.1 equivalents) was added in three equal lots at 0°C, and the mixture was stirred at 25°C for 1 hour. To the resulting mixture, SEM-Cl (382.3 mg, 2.29 mmol, 1.01 equivalents) was added dropwise at 0°C. The resulting reaction mixture was stirred under an argon atmosphere at 25°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (yellow suspension) was quenched by adding ice-cold water (15 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (350 mg). This was purified by flash column chromatography using 20-25% ethyl acetate in hexane as the eluent to obtain product B as a pale yellow oil.

[0174] Yield: 250 mg, 46% 1H-NMR (DMSO-d6,300MHz)δ:8.57(1H,d,J=4.5Hz), 7.97(1H,s), 5.43(2H,s), 3.56-3.52(2H,m), 2.37(3H,s), 0.85(2H,t,J=3.3Hz), -0.01(9H,s) LCMS:241.1[M+H] +

[0175] Step 2: Synthesis of tert-butyl(E)-3-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-pyrazolyl)-2-butenoate(D): [ka]

[0176] Compound C (0.839 g, 3.33 mmol, 2 equivalents) was added dropwise to a stirred suspension of sodium hydride (60% of mineral oil content) (140 mg, 3.33 mmol, 2 equivalents) in THF (6 mL) at 0°C, and the mixture was stirred in an ice bath for 30 minutes. Compound B (400 mg, 1.66 mmol, 1 equivalent) was added dropwise to the resulting suspension. The resulting reaction mixture was gradually warmed to ambient temperature and stirred under an argon atmosphere for 16 hours. The progress of the reaction was monitored by TLC.

[0177] The reaction mixture (yellow suspension) was quenched by adding ice-cold water (10 mL) and extracted with ethyl acetate (3 × 15 mL). The combined organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure to obtain the crude product (350 mg). The crude product was purified by flash column chromatography using 5-10% ethyl acetate in hexane as the eluent to obtain product D.

[0178] Yield: 280 mg as a yellow viscous liquid, 49.7%. 1H-NMR (DMSO-d6,300MHz)δ:8.38-8.28(1H,m), 7.92-7.88(1H,m), 6.08(1H,s), 5.40-5.35(2H,m) , 3.52(2H,t,J=7.8Hz), 2.37-2.41(3H,m), 1.43-1.40(9H,s), 0.82(2H,t,J=8.1Hz), -0.02(9H,s) LCMS:339.0[M+H] +

[0179] Step 3: Synthesis of (E)-3-(4-pyrazolyl)-2-butenoic acid (E): [ka]

[0180] A solution of compound C (280 mg, 0.82 mmol, 1 equivalent) in 1,4-dioxane with 4 M HCl (6 mL) was stirred at room temperature for 48 hours. The reaction mixture (white suspension) was evaporated at 45°C to remove excess HCl and dried. The resulting semi-solid residue was pulverized with diethyl ether (5 mL x 2) to obtain the HCl salt of product E as a white solid.

[0181] Yield: 110 mg, 87.4% 1 H-NMR(DMSO-d6,400MHz)δ:8.00(2H,s), 6.13(1H,s), 2.39(3H,s); LCMS: 152.9 [M+H] + ,HPLC:purity 94.21%

[0182] Example 2 Synthesis of -(E)-3-(2-amino-1,3-thiazole-4-yl)acrylic acid (D) (Compound 2) Step 1: Synthesis of (E)-3-[2-(tert)-butoxycarbonylamino)-1,3-thiazole-4-yl]acrylic acid (C): [ka]

[0183] To a stirred solution of (4-formyl-thiazole-2-yl)-carbamate tert-butyl ester (A) (200 mg, 0.877 mmol, 1 equivalent) in pyridine (4 mL), malonic acid (B) (104 mg, 1.01 mmol, 1.15 equivalents) and piperidine (11.2 mg, 0.131 mmol, 0.15 equivalents) were added under an argon atmosphere at ambient temperature. The resulting reaction mixture was stirred in a sealed container at 90°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture was evaporated under reduced pressure to obtain the crude product (300 mg). The crude product was then purified by grinding with MTBE to obtain product C.

[0184] Yield: 200 mg, 84.5% 1 H-NMR (DMSO-d6,300MHz)δ:7.35(1H,s), 7.18(1H,d,J=14.7Hz,), 6.36(1H,d,J=15.6Hz,d), 1.47(9H,s) LCMS: 269.0 [MH] -

[0185] Step 2: Synthesis of (E)-3-(2-amino-1,3-thiazole-4-yl)acrylic acid (D): [ka]

[0186] To a stirred solution of compound C (100 mg, 0.370 mmol, 1 equivalent) in DCM (1 mL), TFA (1 mL) was added dropwise at 0°C. The resulting reaction mixture was stirred under an argon atmosphere at 25–30°C for 16 hours. The progress of the reaction was monitored by TLC. After the reaction was complete, the white suspension was evaporated under reduced pressure, and the residue was pulverized with diethyl ether (2 mL) to obtain the crude product (80 mg). The crude product was purified by RP-HPLC to obtain product D.

[0187] Yield: 55 mg, 87.3% (CFB-34) was obtained as an off-white solid. 1H-NMR (DMSO-d6,300MHz)δ:7.90(2H,bs), 7.27(1H,d,J=15.6Hz), 7.14(1H,s), 6.26(1H,d,J=15.6Hz) LCMS:171.0[M+H] + ,HPLC:purity 99.96%

[0188] Example 3 Synthesis of -(E)-3-(2-amino-4-imidazolyl)acrylic acid (F) (Compound 3) Step 1: Synthesis of {2-[(4-bromo-2-nitro-1-imidazolyl)methoxy]ethyl}tris(methyl)silane (B) and its isomer (i.e., {2-[(5-bromo-2-nitro-1-imidazolyl)methoxy]ethyl}tris(methyl)silane): [ka]

[0189] To a stirred solution of compound A (1.5 g, 7.89 mmol, 1 equivalent) in THF (40 mL), sodium hydride (60% in mineral oil) (0.350 g, 8.68 mmol, 1.1 equivalents) was added gradually at 0°C. The resulting reaction mixture was stirred at 22°C for 1 hour. After 1 hour, SEM-Cl (1.46 mL, 8.28 mmol, 1.05 equivalents) was added dropwise over 10 minutes at 0°C. The reaction mixture was then stirred under an argon atmosphere at ambient temperature for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (brown solution) was quenched by adding an ice-cold solution of saturated NH4Cl (100 mL), and the product was extracted in ethyl acetate (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (2.3 g). The crude substance was purified by flash column chromatography using 10-15% ethyl acetate in hexane as the eluent to obtain compound B and its isomers as colorless viscous liquids.

[0190] Yield: 1.6g, 63.6% (mixture of isomers) 1H-NMR (CDCl3,400MHz)δ: (mixture of isomers): 7.30-7.21(1H,m), 5.88-5.77(2H,m), 3.70-3.72(2H,m), 0.99-0.90(2H,m), 0.07-0.02(9H,m) LC-MS: No ionization.

[0191] Step 2: Synthesis of 4-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}-2-imidazolylamine (C) and its isomers: [ka]

[0192] Compound B and its isomer (3 g, 9.3 mmol, 1 equivalent) in THF (30 mL) and (2% TPGS-750M) in water (12 mL) were stirred together. NH4Cl (1.50 g, 27.9 mmol, 3 equivalents) was added, followed by the addition of iron carbonyl powder (2.5 g, 46.5 mmol, 5 equivalents). The resulting reaction mixture was vigorously stirred under an argon atmosphere at 40°C for 24 hours. The progress of the reaction was monitored by TLC. The reaction mixture (yellow solution and black suspension) was filtered through a Celite bed. The filtrate was diluted with water (50 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and then concentrated under reduced pressure to obtain a mixture of compound C and its isomers as a pale yellow viscous oil.

[0193] Yield: 2g of crude material: (mixture of isomers) 1 H-NMR (DMSO, 400MHz) δ: (mixture of isomers) 6.71(1H,s), 5.75-5.73(2H,m), 5.06-5.01(2H,m), 3.47-3.742(2H,m), 0.83-0.77(2H,m), -0.04-0.06(9H,m) LCMS:294.1[M+H] +

[0194] Step 3: Synthesis of (4-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}-2-imidazolyl)-N-tert-butoxycarbonylamino-tert-butylformate (D): [ka]

[0195] To a stirred solution of compound C and its isomer (1 g, 3.42 mmol, 1 equivalent) in THF (10 mL), DIPEA (1.10 g, 8.56 mmol, 2.5 equivalents), DMAP (83 mg, 0.68 mmol, 0.2 equivalents), and anhydrous BOC (1.85 g, 8.55 mmol, 2.5 equivalents) were successively added at 0°C. The resulting reaction mixture was stirred under an argon atmosphere at ambient temperature for 16 hours. The reaction mixture (yellowish solution) was added to 100 mL of purified water and extracted with ethyl acetate (2 × 100 mL). The combined organic layer was dried on anhydrous sodium sulfate and then concentrated under reduced pressure to obtain the crude product (1.3 g). The crude substance was purified by flash column chromatography using 30% ethyl acetate in hexane as the eluent to obtain compound D as a yellow solid.

[0196] Yield: 700 mg, 42% 1 H-NMR (DMSO-d6,400MHz)δ:7.49(1H,s), 5.12(2H,s), 3.46-3.42(2H,m), 1.38-1.36(18H,m), 0.87-0.83(2H,m), -0.033(9H,s) LCMS:492[M+H] + .

[0197] Step 4: Synthesis of tert-butyl(E)-3-(2-amino-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-imidazolyl)acrylate(E): [ka]

[0198] A stirred solution of compound D (500 mg, 1.02 mmol), TEA (283 mL, 2.03 mmol, 2 equivalents), and t-butyl acrylate (652 mg, 5.1 mmol, 5 equivalents) in DMF (5 mL) was degassed with argon for 15 minutes. Pd(PPh3)2Cl2 (63.72 mg, 0.051 mmol, 0.05 equivalents) was added to the reaction mixture and degassed further with argon for 5 minutes. The resulting reaction mixture was stirred at 120°C for 48 hours under argon atmosphere and in a sealed condition. The progress of the reaction was monitored by TLC. The reaction mixture (brown solution) was concentrated under reduced pressure. The resulting residue was suspended in 50 mL of purified water and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (600 mg). The crude substance was purified by flash column chromatography using 5-8% MeOH in DCM, followed by RP HPLC, to obtain compound E as a light brown solid.

[0199] Yield: 4 mg, 0.9% 1 H-NMR(CD3OD,400MHz)δ:7.25(1H,d,J=15.6Hz), 7.02(1H,s), 6.15(1H,d,J=15.2Hz) , 5.13(2H,s), 3.56(2H,t,J=8.2Hz), 1.48(9H,bs), 0.91-0.87(2H,m), -0.023(9H,s) LCMS:340.3[M+H] +

[0200] Step 5: Synthesis of (E)-3-(2-amino-4-imidazolyl)acrylic acid (F): [ka]

[0201] A solution of compound E (7 mg, 0.020 mmol, 1 equivalent) in 4N HCl in dioxane (2 mL) was stirred at ambient temperature for 4 days under sealed conditions. The progress of the reaction was monitored by TLC. The reaction mixture (white suspension) was concentrated under reduced pressure to obtain the crude product (6 mg). The obtained crude product was diluted with purified water (5 mL), and the aqueous layer was washed with diethyl ether (3 mL x 2). The aqueous layer was concentrated to 2 mL under reduced pressure and freeze-dried to obtain product F as an off-white semi-solid.

[0202] Yield: 3 mg LCMS: 154.06 [M+H] +

[0203] Example 4 Synthesis of -(E)-3-(4-pyrazolyl)-2-pentenoic acid (G) (compound 4) Step 1: Synthesis of 1-{[2-(trimethylsilyl)ethoxy]methyl}-4-pyrazolecarboxylic acid (B): [ka]

[0204] To a stirred solution of compound A (2.5 g, 0.022 mol, 1 equivalent) in THF (25 mL), sodium hydride (60% in mineral oil) (2.81 g, 0.0669 mol, 3 equivalents) was added in four equal lots at 0–5°C and stirred at the same temperature for 1 hour. After 1 hour, SEM-Cl (4.14 mL, 0.023 mol, 1.05 equivalents) was added dropwise at 0–5°C. The resulting reaction mixture was stirred under an argon atmosphere at 25–30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (gray suspension) was quenched by adding ice-cold water (25 mL), acidified to approximately pH 4 with 1.5 N HCl, and then extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (3.8 g). The crude substance was purified by flash column chromatography using 20-23% ethyl acetate in hexane as the eluent, and product B was obtained as a white solid.

[0205] Yield: 3.5g, 63.0% 1 H-NMR (CDCl3,400MHz)δ:8.14(1H,s), 8.01(1H,s), 5.47(2H,s), 3.63-3.59(2H,m), 0.96-0.92(2H,m), -0.04(9H,s). LCMS:245.1[M+H] +

[0206] Step 2: Synthesis of N-methoxy-N-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-pyrazolecarboxamide (C): [ka]

[0207] To a stirred solution of compound B (3 g, 0.0124 mol, 1 equivalent) in DCM (60 mL) at 0-5°C, TEA (3.7 g, 0.0186 mol, 3 equivalents), EDC.HCl (3.5 g, 0.0186 mol, 1.5 equivalents), and HOBt (0.95 g, 0.0062 mol, 0.5 equivalents) were successively added, and the mixture was stirred at the same temperature for 5 minutes. N-dimethylhydroxylamine (1.45 g, 0.014 mol, 1.2 equivalents) was added to the reaction mixture in one lot. The resulting reaction mixture was stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (yellow solution) was quenched by adding ice-cold water (50 mL) and extracted into DCM (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and then concentrated under reduced pressure to obtain the crude product (3.5 g). The crude product was purified by flash column chromatography using 15-20% ethyl acetate in hexane as the eluent to obtain product C as a pale yellow viscous liquid.

[0208] Yield: 2.7g, 76.2% 1H-NMR (CDCl3,400MHz)δ:8.12(1H,s), 8.02(1H,s), 5.45(2H,s), 3.73(3H,s), 3.61-3.57(2H,m), 3.34(3H,s), 0.92(2H,t,J=8.4Hz), -0.02(9H,s) LCMS:286.2[M+H] +

[0209] Step 3: Synthesis of 1-(1-{[2-(trimethylsilyl)ethoxy]methyl}-4-pyrazolyl)-1-propanone (D): [ka]

[0210] To a solution of compound C (2.5 g, 0.0087 mol, 1 equivalent) in dry THF (50 mL), ethylmagnesium bromide (5.8 g, 0.0437 mol, 5 equivalents) was added dropwise at 0°C. The resulting reaction mixture was stirred under an argon atmosphere at ambient temperature for 5 hours. The progress of the reaction was monitored by TLC. The reaction mixture (yellow solution) was quenched by adding saturated NH4Cl solution (50 mL) and extracted in ethyl acetate (3 × 50 mL). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (2.8 g). The crude substance was purified by flash column chromatography using 15-20% ethyl acetate in hexane as the eluent to obtain the product as a pale yellow viscous liquid.

[0211] Yield: 1.8g, 81.1% 1 H-NMR(CDCl3,400MHz)δ:8.06(1H,s), 7.96(1H,s), 5.45(2H,s), 3.61-3.57(2H,m ), 2.84(2H,q,J=7.2Hz), 1.21(3H,t,J=7.6Hz), 0.95-0.91(2H,M), -0.01(9H,s). LCMS:255.1[M+H] +

[0212] Step 4: Synthesis of tert-butyl(E)-3-(1-{[2-(methylsilyl)ethoxy]methyl}-4-pyrazolyl)-2-pentenoate(F): [ka]

[0213] To a solution of NaH (60% in mineral oil) (1.24 g, 0.031 mol, 5 equivalents) in THF (13 mL), tert-butyl(E) diethylphosphonoacetate (7.9 g, 0.031 mol, 5 equivalents) was added at 0°C, and the mixture was stirred at 0-5°C for 20 minutes. Compound D (1.6 g, 0.0062 mol, 1 equivalent) in THF (5 mL) was added to this mixture at 0-5°C, and the resulting reaction mixture was stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (yellow) was quenched by adding ice-cold water (25 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (1.8 g). The crude substance was purified by flash column chromatography using 20-23% ethyl acetate in hexane as the eluent to obtain compound F as a pale yellow viscous liquid.

[0214] Yield: 1.1g, 45% 1 H-NMR(CDCl3,400MHz)δ:8.06(1H,s), 7.96(1H,s), 5.45(2H,s), 3.61-3.57(2H,m ), 2.84(2H,q,J=7.2Hz), 1.21(3H,t,J=7.6Hz), 0.95-0.91(2H,M), -0.01(9H,s); LCMS:353.3[M+H] +

[0215] Step 5: Synthesis of (E)-3-(4-pyrazolyl)-2-pentenoic acid (G): [ka]

[0216] To a solution of compound F (700 mg, 0.0019 mol, 1 equivalent) in 1,4-dioxane (3.5 mL), 4 M HCl (3.5 mL) was added to the 1,4-dioxane. The resulting reaction mixture was stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (white suspension) was evaporated at 45°C to remove excess HCl and dried. The resulting semi-solid residue was pulverized with diethyl ether (5 mL x 2) to obtain product G as a white solid.

[0217] Yield: 55 mg, 16.7% 1 H-NMR (DMSO-d6,400MHz)δ:7.99(2H,s), 6.05(1H,s), 2.93(2H,q,J=7.6Hz), 1.07(3H,t,J=7.2Hz) LCMS: 167.0 [M+H] - ,HPLC:purity 95.6%

[0218] Example 5 Synthesis of -(E)-3-(4-imidazolyl)-2-butenoic acid (F) (compound 5) Step 1: Synthesis of 1-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-imidazolyl)-1-ethanone (B) and its isomers: [ka]

[0219] To a stirred solution of compound A (2 g, 0.0181 mol, 1 equivalent) in THF (20 mL), sodium hydride (60% in mineral oil) (0.79 g, 0.0199 mol, 1.1 equivalents) was added in four equal lots at 0°C and stirred at 25°C for 1 hour. After 1 hour, SEM-Cl (3.17 g, 0.019 mol, 1.05 equivalents) was added dropwise at 0°C. The resulting reaction mixture was then stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (gray suspension) was quenched by adding ice-cold water (20 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (2.8 g). The crude substance was purified by flash column chromatography using 20-23% ethyl acetate in hexane as the eluent to obtain product B and its isomers as white solids.

[0220] Yield: 2.3g, 52.7% (mixture of isomers) 1 H-NMR (CDCl3, 400MHz) δ: (mixture of isomers) 7.69 (1H,d,J=1.2Hz), 7.61 (1H,d,J=1.2H z), 5.31(2H,s), 3.50(2H,t,J=8.2Hz), 0.95-0.90(2H,m), 0.02--0.01(9H,m). LCMS: 240.90 [M+H] +

[0221] Step 2: Synthesis of ethyl(E)-3-(1-{[2-(trimethylsilyl)ethoxy]methyl}-4-imidazolyl)-2-butenoate(D): [ka]

[0222] To a stirred solution of compound C (691.93 mg, 2.0833 mmol) in THF at -75°C, KHMDS (1 M in THF) (2.18 mL, 2.1874 mmol) was added dropwise while maintaining the internal temperature at <-65°C, and the mixture was then stirred for 30 minutes. To the above reaction mixture, compound B and its isomers (250 mg, 1.04166 mmol) in THF (2 mL) were added dropwise while maintaining the internal temperature at <-65°C. The resulting reaction mixture was gradually warmed to ambient temperature within 1.5 hours and stirred at ambient temperature for 18 hours. The progress of the reaction was monitored using TLC. The resulting reaction mixture was quenched by adding saturated NH4Cl aqueous solution (5 mL) at 0°C and extracted with ethyl acetate (3 × 15 mL). The combined organic layers were dried over sodium sulfate and concentrated at 40°C to obtain the crude product, which was then purified by flash column to obtain compound D and its isomers as a concentrated oily yellow liquid.

[0223] Yield: 90 mg (27.8%) (mixture of isomers) 1 H-NMR (CDCl3,300MHz) δ: (mixture of isomers) 8.30(1H,s), 7.76(1H,s), 7.60(1H,s), 6.69(1H,s), 5.88(1H,s), 5.33(2H,s), 5.27(2H,s ), 4.23-4.14(2H,m), 3.57-3.48(2H,m), 2.51(3H,s), 2.38(3H,s), 1.28(6H,t,J=7.2Hz), 0.93(4H,t,J=8.1Hz), 0.01(18H,s) LCMS:311.2[M+H] +

[0224] Step 3: Synthesis of (E)-3-(1-{[2-(trimethylsilyl)ethoxy]methyl}-4-imidazolyl)-2-butenoic acid (E): [ka]

[0225] To a stirred solution of compound D and its isomer (90 mg, 0.2898 mmol) in THF:ethanol:water (1 mL, 1:1:1 equivalents), lithium hydroxide monohydrate (30.42 mg, 0.7245 mmol) was added. The resulting reaction mixture was stirred at 25-30°C for 18 hours. The progress of the reaction was monitored by TLC. After 18 hours, the reaction mixture was concentrated, the residue was dissolved in water, and the aqueous layer was acidified to approximately pH 5-6 using 1.5N HCl. The solution was extracted with ethyl acetate (3 × 10 mL), the combined organic layers were dried over sodium sulfate, and concentrated at 40°C to obtain the crude substance. The crude substance was washed with hexane (5 mL) to obtain crude compound E and its isomer as a concentrated oily yellow liquid.

[0226] Yield: 55 mg (mixture of isomers) 1 H-NMR (CDCl3,300MHz)δ: (mixture of isomers) 7.92(1H,s), 7.79(1H,s), 6.78(1H,s), 5.99(1H,s), 5 .35(4H,s), 3.57-3.52(4H,m), 2.52(3H,s), 2.20(3H,s), 1.79-1.70(4H,m), 0.01(18H,s) LCMS:283.2[M+H] +

[0227] Step 4: Synthesis of (E)-3-(4-imidazolyl)-2-butenoic acid (F): [ka]

[0228] To a stirred solution of pooled compound E and its isomer (1.7 g, 0.0060 mmol) in 1,4-dioxane (15 mL), 4 M HCl in 1,4-dioxane was added (34 mL). The resulting reaction mixture was stirred at 25-30°C for 18 hours. The progress of the reaction was monitored by TLC. The reaction mixture was evaporated under reduced pressure, followed by toluene chasing to obtain the crude product (1.2 g). This was purified by RP-HPLC and dried by lyophilization to obtain compound F (fraction 1) and its Z isomer (fraction 2). Fraction 1 was further purified by sonication with water (10 mL) and filtered. This yielded the expected product F.

[0229] Yield: 150 mg 1 H-NMR(CD3OD,400MHz)δ:7.71(1H,d,J=0.8Hz), 7.41(1H,d,J=0.8Hz), 6.41(1H,d,J=1.2Hz), 2.28(3H,s) LCMS:153.0[M+H] + , HPLC purity 99.96%

[0230] Example 6 Synthesis of -(E)-3-(5-methyl-4-pyrazolyl)-2-butenoic acid (E) (compound 6) Step 1: Synthesis of 1-(5-ethyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-pyrazolyl)-1-ethanone (B) and its isomers: [ka]

[0231] To a stirred solution of compound A (230 mg, 2.416 mmol, 1 equivalent) in THF (5 mL), sodium hydride (57% in mineral oil) (122.1 mg, 2.899 mmol, 1.2 equivalents) was added at 0°C in two equal lots, and the mixture was stirred at 25°C for 1 hour. After 1 hour, SEM-Cl (443.13 mg, 2.658 mmol, 1.1 equivalents) was added dropwise at 0°C. The resulting reaction mixture was stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC.

[0232] The reaction mixture (gray suspension) was quenched by adding ice-cold water (10 mL) and extracted with ethyl acetate (3 × 15 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (520 mg). The crude product was purified by flash column chromatography using 30-40% ethyl acetate in hexane as the eluent to obtain product B and its isomers as white solids.

[0233] Yield: 320 mg, 52.2% (mixture of isomers) 1 H-NMR (CDCl3,400MHz)δ: (mixture of isomers) 7.96(1H,s), 7.82(1H,s), 5.44(2H,s), 5.36(2H,s), 3.62-3.54(4H,m), 2.6 3(3H,s,), 2.48(3H,s), 2.44(3H,s), 2.43(3H,s), 2.42(3H,s), 0.94-0.87(4H,m), -0.01(9H,s,), -0.03(9H,s) LCMS:255.03[M+H] +

[0234] Step 2: Synthesis of tert-butyl(E)-3-(5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-pyrazolyl)-2-butenoate(D): [ka]

[0235] Compound C (1.49 g, 5.895 mmol, 5 equivalents) was added at 0°C to a stirred suspension of NaH (57% of mineral oil) (253.1 mg, 6.014 mmol, 5.1 equivalents) in THF (5 mL) and stirred at 0°C for 30 minutes. Compound B and a mixture of its isomers (300 mg, 1.179 mmol, 1 equivalent) were added at 0°C, and the resulting reaction mixture was stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction solution (pale yellow) was quenched by adding ice-cold water (10 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (480 mg). The crude product was purified by flash column chromatography using 10-15% ethyl acetate in hexane as the eluent to obtain product D and its isomers as a pale yellow viscous liquid.

[0236] Yield: 315 mg, 75.8% (mixture of isomers) 1 H-NMR (CDCl3,400MHz) δ: (mixture of isomers) 7.59(1H,s), 7.53(1H,s), 5.91(1H,d,J=1.2Hz), 5.91(1H,d,J=1.2Hz), 5.77(1H,d,J=1.6Hz ), 5.42(2H,s), 5.40(2H,s), 3.58-3.52(4H,m), 2.47-2.39(12H,m), 2.57-2.38(18H,m), 0.93-0.86(4H,m), -0.01-0.04(18H,m) LCMS:353.4[M+H] +

[0237] Step 3: Synthesis of (E)-3-(5-methyl-4-pyrazolyl)-2-butenoic acid (E): [ka]

[0238] Compound D and its isomer (300 mg, 1.477 mmol, 1 equivalent) were stirred in 1,4-dioxane (3 mL). 4M HCl (6 mL) was added to the 1,4-dioxane, and the resulting reaction mixture was stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (white suspension) was evaporated under reduced pressure, and volatiles were removed at 45°C to obtain the crude product, which was further purified by RP-HPLC to obtain two fractions of equal mass. The fractions from which the isomers were recovered were evaporated at 45°C, and the resulting residue was dissolved in ACN:water (1:2) (2 mL) and dried by freeze-drying to obtain the desired product E as fraction 2 (58 mg) (off-white solid).

[0239] Yield of compound E: 58 mg, 23.7% 1 H-NMR(CD3OD,400MHz)δ:7.72(1H,s), 5.93(1H,d,J=1.2Hz), 2.47(3H,d,J=1.2Hz), 2.38(3H,s) LCMS: 167.0 [M+H] + ,HPLC:purity 99.56%

[0240] Example 7 Synthesis of -(E)-3-(3-pyrrolyl)-2-butenoic acid (F) (compound 7) Step 1: Synthesis of 1-(1-tosyl-3-pyrrolyl)-1-ethanone (B): [ka]

[0241] To a stirred solution of compound A (0.5 g, 0.0046 mol, 1 equivalent) in DCM (5 mL), TEA (1.16 g, 0.0115 mol, 2.5 equivalents) and DMAP (28 mg, 0.0002 mol, 0.05 equivalents) were added at ambient temperature. To this reaction mixture, p-toluenesulfonyl chloride (1.13 g, 0.0059 mol, 1.3 equivalents) was added. The resulting reaction mixture was stirred under an argon atmosphere at ambient temperature for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (white suspension) was quenched by adding ice-cold water (10 mL) and extracted with ethyl acetate (3 × 25 mL). The combined organic layers were dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (0.6 g). The crude product was purified by flash column chromatography using 20-30% ethyl acetate in hexane as the eluent to obtain compound B as a white solid.

[0242] Yield: 0.59g, 48.7% 1 H-NMR(CDCl3,400MHz)δ:7.79(2H,q,J=1.24Hz), 7.71(1H,q,J=1.33Hz), 7.34(2H, t,J=2.8Hz), 7.13(1H,q,J=1.8Hz), 6.67(1H,q,J=1.6Hz), 2.43(3H,s), 2.40(3H,s) LCMS:264.0[M+H] +

[0243] Step 2: Synthesis of tert-butyl(E)-3-(1-tosyl-3-pyrrolyl)-2-butenoate(D): [ka]

[0244] Compound C (0.99 g, 0.0026 mol, 1.2 equivalents) was added in one lot to a stirred solution of compound B (0.58 g, 0.0022 mol, 1 equivalent) in ACN (2.32 mL). The resulting reaction mixture was stirred at 85°C for 48 hours under an argon atmosphere. The progress of the reaction was monitored by TLC. The reaction mixture (brown solution) was quenched by adding ice-cold water (10 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product (0.62 g). The crude product was purified by flash column chromatography using 5-10% ethyl acetate in hexane as the eluent to obtain compound D as a pale yellow viscous liquid.

[0245] Yield: 0.33g, 41.5% 1 H-NMR(CDCl3,400MHz)δ:7.78-7.74(2H,m), 7.33-7.29(3H,m), 7.12(1H,t,J=1.6 Hz), 6.59-6.47(1H,m), 5.98(1H,d,J=1.2Hz), 2.39(6H,q,J=3.2Hz), 1.48(9H,s) LCMS:384.2[M+Na] +

[0246] Step 3: Synthesis of (E)-3-(1-tosyl-3-pyrrolyl)-2-butenoic acid (E): [ka]

[0247] Compound D (320 mg, 0.885 mmol, 1 equivalent) was stirred in 1,4-dioxane (1.6 mL). 4M HCl (12.8 mL) was added to the 1,4-dioxane, and the resulting reaction mixture was stirred under an argon atmosphere at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (white suspension) was evaporated and dried under reduced pressure at 45°C to obtain compound E as a white solid.

[0248] Yield: 230 mg, 85.1% 1 H-NMR(DMSO,400MHz)δ:7.88(2H,d,J=8.4Hz), 7.67(1H,d,J=2.0Hz), 7.43(2H, d,J=8Hz), 7.33(1H,t,J=Hz), 6.69(1H,t,J=Hz), 6.07(1H,s), 2.35-2.30(6H,m) LCMS:304.1[M+H] - .

[0249] Step 4: Synthesis of (E)-3-(3-pyrrolyl)-2-butenoic acid (F): [ka]

[0250] To a stirred solution of compound E (220 mg, 0.721 mmol, 1 equivalent) in THF (2.2 mL) and methanol (4.4 mL), Cs2CO3 (704.2 mg, 2.164 mmol, 3 equivalents) was added, and the resulting reaction mixture was stirred at 50°C for 18 hours under an argon atmosphere. The progress of the reaction was monitored by TLC. The reaction mixture (yellow suspension) was concentrated under reduced pressure at 25-30°C to obtain the crude product (350 mg). The crude product was purified by RP-HPLC and dried by lyophilization (without using a rotary evaporator for product sensitivity). This yielded product F as an off-white solid.

[0251] Yield: 66 mg, 26.9% 1 H-NMR(CD3OD,400MHz)δ:7.06(1H,t,J=1.6Hz), 6.71(1H,q,J=1.6Hz), 6.35(1H,q,J=1.6Hz), 6.05(1H,d,J=1.2Hz), 2.42(3H,d,J=0.8Hz) LCMS: 149.7[M+H] - ,HPLC:purity 94.1%

[0252] Example 8 Synthesis of -(E)-3-(2-amino-4-imidazolyl)-2-butenoic acid (H) (compound 8) Step 1: Synthesis of ethyl(Z)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-butenoate(B): [ka]

[0253] A stirred suspension of ethyl porcine-2-isoate (A) (2 g, 17.86 mmol) in water (200 mL) was degassed with nitrogen for 10 minutes. CuSO4 (142 mg, 0.89 mmol), 4-methylpyridine (415 mg, 4.47 mmol), and bis(pinacorato)diborone (B2pin2) (3.40 g, 13.40 mmol) were successively added to this solution, and the resulting reaction mixture was stirred at 50°C for 10 minutes. A second batch of B2pin2 (3.40 g, 13.40 mmol) was added to the resulting solution, and the progress of the reaction was monitored by TLC. After 16 hours, the reaction mixture was diluted by adding hexane. The organic phase was separated, and the aqueous phase was extracted with hexane (150 mL x 3). The combined organic layers were washed with water and dried over Na2SO4. The solvent was evaporated under reduced pressure at 45°C, and the crude residue was purified by Combiflash® chromatography using 8-10% ethyl acetate in hexane. The recovered fraction was concentrated under reduced pressure to obtain compound B as a colorless, concentrated oil.

[0254] Yield 1.1g, 52% 1 H-NMR(CDCl3,400MHz)δ:6.45(1H,d,J=1.6Hz), 4.17(2H,q,J=7.2Hz), 2.17(3H,s,), 1.29-1.26(15H,m)

[0255] Step 2: Synthesis of {2-[(4-bromo-2-nitro-1-imidazolyl)methoxy]ethyl}tris(methyl)silane (D) or its isomer (i.e., {2-[(5-bromo-2-nitro-1-imidazolyl)methoxy]ethyl}tris(methyl)silane) [ka]

[0256] To a stirred solution of 4-bromo-2-nitro-1H-imidazole (C) (300 mg, 1.57 mmol) in THF (15 mL), sodium hydride (60% in mineral oil) (70 mg, 1.74 mmol) was gradually added at 0°C. The resulting reaction mixture was stirred at 22°C for 1 hour. Then, SEM-Cl (2-(trimethylsilyl)ethoxymethyl chloride) (325 mg, 1.65 mmol) was added dropwise over 10 minutes at 0°C. The reaction mixture was stirred under an argon atmosphere at ambient temperature for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture (brown solution) was quenched by adding an ice-cold solution of saturated NH4Cl (75 mL), and the product was extracted in ethyl acetate (3 × 50 mL). The combined organic phase was dried over anhydrous sodium sulfate and then concentrated under reduced pressure to obtain the crude product (530 mg). This was purified by flash column chromatography using 10-15% ethyl acetate in hexane as the eluent to obtain compound D or its isomer as a colorless viscous liquid.

[0257] Yield of D or its isomer: 260 mg (50%) 1 H-NMR (CDCl3,400mHz) δ (D or its isomer): 7.29 (1H,s), 5.76 (2H,s), 3.67 (2H,t,J=8.4Hz), 0.98 (2H,t,J=8.4Hz), 0.02 (9H,s).

[0258] Step 3: Synthesis of ethyl(E)-3-(2-nitro-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-imidazolyl)-2-butenoate(E) or its isomers: [ka]

[0259] A stirred solution of compound D or its isomer (600 mg, 1.86 mmol) and compound B (895 mg, 3.72 mmol) in 1,4-dioxane (10 mL) was degassed with argon for 5 minutes. Pd(dppf)Cl was added to this solution.2· CH2Cl2 complex (151 mg, 0.186 mmol) and K2CO3 (1.02 g, 7.44 mmol) were added while degassing for 5 minutes. The resulting reaction mixture was then stirred at 90°C under an argon atmosphere for 12 hours. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with water and extracted with pharmaceutically acceptable (3 × 20 mL) HCl. The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The resulting crude product was purified by flash chromatography (30% HCl in hexane) to obtain compound E or its isomer as a yellow solid.

[0260] Yield of E or its isomer: 328 mg (50%) 1 H-NMR (CDCl3,400mHz)δ (E or its isomer): 7.45(1H,s), 6.72(1H,d,J=1.6Hz), 5.76(2H,s), 4.23-4.16(2 H,m), 3.68-3.64(2H,m), 2.51(3H,d,J=1.6Hz), 1.32-1.23(3H,m), 0.99-0.98(2H,m), 0.01(9H,s). LCMS:378.3[M+Na] +

[0261] Step 4: Synthesis of ethyl(E)-3-(2-amino-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-imidazolyl)-2-butenoate(F) or its isomer: [ka]

[0262] To a solution of compound E or its isomer (280 mg, 0.789 mmol) in THF (10 mL), Zn (280 mg, w / w) and NH4Cl (280 mg) in water (3 mL) were successively added, and the mixture was stirred at ambient temperature for 24 hours. The progress of the reaction was monitored by TLC. After the starting material was completely consumed, the reaction mixture was diluted with ethyl acetate (50 mL) and filtered through a Celite bed. The organic layer was separated and concentrated to obtain compound F or its isomer as an off-white solid.

[0263] Yield of F or its isomer: 163 mg (63%). 1 H-NMR (CDCl3,400mHz)δ (F or its isomer): 6.80(1H,s), 6.49(1H,d,J=1.2Hz), 5.09(2H,s), 4.48(2H,bs), 4.19 -4.14(2H,m), 3.56-3.51(2H,m), 2.40(3H,d,J=1.2Hz), 1.29-1.24(3H,m), 0.94-0.90(2H,m), 0.0(9H,s) LCMS:326.4[M+H] +

[0264] Step 5: Synthesis of (E)-3-(2-amino-1-{[2-(trimethylsilyl)ethoxy]methyl}-4-imidazolyl)-2-butenoic acid (G) or its isomer: [ka]

[0265] A stirred solution of compound F or its isomer (150 mg, 0.462 mol) in THF (10 mL) was mixed with a solution of LiOH (55 mg, 2.31 mmol) in water (3 mL) at room temperature. The resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure, and the resulting residue was suspended in water (13 mL) and acidified to approximately pH 4 with 1.5 N HCl. The resulting precipitate was collected by filtration and dried under high vacuum to obtain compound G or its isomer as an off-white solid.

[0266] Yield of G or its isomer: 106 mg (77%) 1 H-NMR (DMSO, 400 mHz) δ (G or its isomer): 11.93 (1H, bs), 6.80 (1 H, s), 6.49 (1 H, d, J=1.2 Hz), 5.15(2H,s), 3.53-4.48(2H,m), 2.25(3H,d,J=1.2Hz), 0.86-0.84(2H,m), -0.03(9H,s) LCMS:298.3[M+H] +

[0267] Step 6: Synthesis of (E)-3-(2-amino-4-imidazolyl)-2-butenoic acid (H): [ka]

[0268] Compound G or its isomer (100 mg, 0.337 mmol) was stirred in 1 mL of 1,4-dioxane, to which 1 mL of 4 M HCl in 1,4-dioxane was added. The resulting reaction mixture was stirred under an argon atmosphere and in a sealed condition at 25-30°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture was evaporated under reduced pressure, and volatiles were removed at 45°C. The remaining crude product was further purified by RP-HPLC. The combined fraction was evaporated at 45°C, and the resulting residue was dissolved in acetonitrile:water (2:1) (3 mL). This solution was dried by freeze-drying to obtain the final product H (as an off-white solid).

[0269] Yield: 22 mg, 39% 1 H-NMR(DMSO,400MHz)δ:11.93(1H,bs), 7.13(1H,s), 6.54(2H,bs), 6.06(1H,s), 2.25(3H,d,J=1.2Hz) LCMS:168.0[M+H] + ;HPLC purity 94.4%

[0270] Example 9 - Urocanate reductase enzyme assay in cell lysates The urocanate reductase (UrdA) enzyme activity of the compounds was determined in cell lysates of the following bacteria that endogenously express the enzyme: Shewanella oneidensis MR-1 (ATCC 700550), Eggerthella lenta (CCUG 17323 AT), Anaerococcus prevotii (CCUG 41932), and Brevibacillus laterosporus (CCUG 52899). Unless otherwise specified, all chemicals were purchased from Sigma-Aldrich.

[0271] Bacterial cell cultures were diluted to 2% in their respective growth media: - S. oneidensis: Trypticase soy yeast extract medium (TSB, Becton Dickinson AB, 0.25% yeast, supplemented with oxyoid), E. lenta: LYBHI-brain-cardiac infusion medium (BHI, oxyoid) supplemented with 0.5% yeast, A. prevoti and B. laterosporus: BHI medium. The cultures were then heated at 30°C for S. oneidensis, and A. prevoti and B. LaterospolsFor E. lenta, cells were maintained under aerobic conditions at 37°C, or under strict anaerobic conditions in a Coy chamber (5% hydrogen, 10% carbon dioxide, and 85% nitrogen) at 37°C until they reached the steady growth phase. For lysate preparation, cells were recovered by centrifugation (4700g, 4°C, 20 min), washed once with buffer (50mM HEPES, 100mM NaCl, Tris pH 7.0), and then resuspended in the same buffer to a final 50-fold concentration for E. lenta cells, or to a final 10-fold concentration for A. prevotiii and B. laterosporus cells. Aliquots were rapidly frozen on dry ice and stored at -80°C until further lysis preparation by glass bead treatment (6 × 1 min at 4°C for E. lenta and A. prevotiii, rested on ice for 3 minutes; B. laterosporus, 4 × 30 seconds at 4°C, rested on ice for 3 minutes). S. oneidensis cells were resuspended to a final 10-fold concentration in a buffer containing 0.025% n-dodecyl-β-d-maltoside and further lysed by sonication (5 × 1 minute in a sonicated water bath, followed by a 30-second pause on ice). Aliquots were rapidly frozen and stored at -80°C until further use.

[0272] The apparent reaction rate parameters were evaluated by nonlinear regression for S. oneidensis, E. lenta, A. prevoti, and B. laterosporus under the assumption of Michaelis-Menten reaction kinetics to determine the appropriate substrate concentrations (corresponding to the Michaelis-Menten constant KM) for each homolog used in subsequent enzyme activity assays.

[0273] The reaction was carried out under anaerobic conditions, at room temperature (for S. oneidensis) and at 37°C (for E. lenta, A. prevoti, and B. laterosporus), in 210 μL of buffer (50 mM HEPES, 100 mM NaCl, Tris pH 7.0) containing the respective urocanic acid concentrations (25 μM urocanic acid for S. oneidensis, 5 μM urocanic acid for E. lenta, 10 μM urocanic acid for A. prevoti, and 0.5 μM urocanic acid for B. laterosporus). 100 mM methyl viologen was reduced with 505 mM sodium dithionite in 100 mM Tris-HCl pH 8 and added to the reaction mixture as an electron donor to a final concentration of 680 μM. Compounds (UrdA inhibitors) were added at different concentrations. The incubation time was adjusted to a linear range of product formation during the preparation of each lysate. The reaction was stopped by removing the sample from the anaerobic environment and adding 210 μL of acetonitrile (UHPLC grade purity, Thermo Fisher) for sample extraction.

[0274] The substrate conversion rate was determined by measuring the concentrations of urocanic acid and imidazole propionate (ImP) using reversed-phase liquid chromatography-tandem mass spectrometry. The extracted sample was then mixed with a 100 nM internal standard substance (ImP- 13 C3 and urocanic acid - 13After mixing with C3 (Astra Zeneca), the sample was dried under a nitrogen stream. The sample was then reconstituted with 5% HCl (37%) in 1-butanol and subjected to n-butyl ester derivatization at 70°C for 40 minutes, and finally reconstituted in water:acetonitrile (9:1). The sample was analyzed using a Waters Acquity UPLC system coupled to a Xevo TQ XS tandem mass spectrometer (Waters) operated in positive electrospray ionization mode. The sample was loaded onto a BEH C18 analytical column (2.1 × 50 mm with 1.7 mm particles; Waters) and separated using a gradient consisting of water containing 0.1% formic acid (phase A) and acetonitrile containing 0.1% formic acid (phase B). The levels of ImP and urocanic acid were determined using multiple reaction monitoring (MRM) of transitions between 197.2>81.2 and 195.2>93.0, according to the method described by Koh et al. (Cell 175(4):947-961 e17, 2018). For the internal standard, the transition between 200.2>82.0 (ImP-- 13 C3) and the transfer of 198.2 > 95.0 (urocanic acid - 13 C3) was used. Enzyme activity was normalized relative to the vehicle control sample, and the substrate conversion rate was maintained within the linear velocity range. IC 50 The values ​​were determined by fitting the normalized data to a nonlinear regression equation with a variable gradient, and are shown in Table 1. [Table 7]

[0275] Example 10 - Urocanic acid reductase enzyme assay using purified protein from Shewanella oneidensis Inhibition of urocanate reductase (UrdA) enzyme activity was determined using purified UrdA (so_4620) from Shewanella oneidensis MR-1. A plasmid for UrdA expression was introduced into Escherichia coli (E. coli) BL21 DE3 cells via electroporation. Gene expression was induced at 0.8 OD600 using 1.0 mmol / L IPTG at 16°C for 16 hours. Cells were harvested by centrifugation, resuspended in buffer (20 mM Tris, pH 7.9, 10% glycerol, 500 mM KCl, 10 mM imidazole, 3 mM DTT), and lysed using 1:1 volume B-PER bacterial protein extraction reagent (Thermo Scientific). The lysate was centrifuged, and the supernatant was taken from Ni Sepharose (商標) The target enzyme was purified using a Fast Flow agarose column, and impurities were removed using 10 mM, 20 mM, and 50 mM imidazole washing solutions. The target enzyme was then eluted with 250 mM imidazole. The eluted protein was desalted using a PD-10 desalting column with elution buffer (20 mM Tris-HCl, 500 mM KCl, 10% glycerol, 3 mM DTT), transferred to an airtight bottle, sealed, and stored at -20°C in 50% glycerol.

[0276] The apparent kinetic parameters were evaluated by nonlinear regression under the assumption of Michaelis-Menten kinetics to determine the appropriate substrate concentration (corresponding to the Michaelis-Menten constant KM) to be used in subsequent enzyme activity assays.

[0277] The reaction was carried out under anaerobic conditions in 210 μL of buffer (50 mM HEPES, 100 mM NaCl, Tris pH 7.0) containing 1.5 μM urocanic acid and 100 μM FAD. 100 mM methyl viologen was reduced with 505 mM sodium dithionite in 100 mM Tris-HCl pH 8 and added to the reaction mixture as an electron donor to a final concentration of 680 μM. Compounds (UrdA inhibitors) were added at different concentrations, followed by the addition of 1 ng of UrdA / reactant and incubation at 37°C for 15 minutes. The reaction was stopped by removing the sample from the anaerobic environment and adding 210 μL of acetonitrile (UHPLC grade purity, Thermo Fisher) for sample extraction.

[0278] The substrate conversion rate was determined by measuring the concentrations of urocanic acid and imidazole propionate (ImP) using reversed-phase liquid chromatography-tandem mass spectrometry. The extracted sample was then mixed with a 100 nM internal standard substance (ImP- 13 C3 and urocanic acid - 13 After mixing with C3 (Astra Zeneca), the sample was dried under a nitrogen stream. The sample was then reconstituted with 5% HCl (37%) in 1-butanol and subjected to n-butyl ester derivatization at 70°C for 40 minutes, and finally reconstituted in water:acetonitrile (9:1). The sample was analyzed using a Waters Acquity UPLC system coupled to a Xevo TQ XS tandem mass spectrometer (Waters) operated in positive electrospray ionization mode. The sample was loaded onto a BEH C18 analytical column (2.1 × 50 mm with 1.7 mm particles; Waters) and separated using a gradient consisting of water containing 0.1% formic acid (phase A) and acetonitrile containing 0.1% formic acid (phase B). The levels of ImP and urocanic acid were determined using multiple reaction monitoring (MRM) of transitions 197.2>81.2 and 195.2>93.0, according to the method described by Koh et al. (Cell 175(4):947-961 e17, 2018). For the internal standard, the transition (ImP-) of 200.2>82.0 was used. 13 C3) and the transfer of 198.2 > 95.0 (urocanic acid -13 C3) was used. Enzyme activity was normalized relative to the vehicle control sample, and the substrate conversion rate was maintained within the linear velocity range. IC 50 The values ​​were determined by fitting the normalized data to a nonlinear regression equation with a variable gradient, and are shown in Table 2. [Table 8]

[0279] Example 11 - Urocanate reductase enzyme assay in intact cells Bacterial cell cultures derived from S. oneidensis were maintained under aerobic conditions at 30°C until they reached the logarithmic growth phase (OD600 = 0.25). The cells were then harvested by centrifugation (4700g, 4°C, 10 min), washed once with buffer (50mM HEPES, 100mM NaCl, Tris pH 7.0), transferred to anaerobic conditions, and resuspended in growth medium (TSBYE) to a final concentration of 0.5 times.

[0280] The reaction was carried out under anaerobic conditions in a total volume of 200 μL containing S. oneidensis cells, 25 μM urocanic acid, and different concentrations of UrdA inhibitors. After incubation at room temperature for 5 minutes, the samples were removed from the anaerobic environment and 210 μL of acetonitrile was added.

[0281] Substrate conversion rate and IC 50 The value was determined using the same method as in Example 10.

[0282] Compound 8 increased the UrdA activity of intact S. oneidensis cells by 0.4 μM IC50. 50 Compound 9 inhibited at 37 μM, confirming the results from the S. oneidensis UrdA lysate assay in Example 10.

[0283] Example 12 - Inhibition of urocanate reductase enzyme activity in multimicrobial human fecal cultures. Materials and methods To determine the efficacy of UrdA inhibitors in a complex environment, UrdA enzyme activity was investigated in multimicrobial human fecal cultures. A modified human gut redox model (SHIRM) was used (Koh et al., Cell, 175(4), 947-961.e17, 2018). The SHIRM model consisted of a two-chamber fermenter with an anaerobic lumen chamber continuously purged with nitrogen and maintained at 37°C, and an oxygen chamber containing 100 mM potassium phosphate buffer. The two compartments were separated by a CMI-7000S membrane (Membrane International, New Jersey). The luminal chamber feed contained (g / L) arabinogalactan (1), pectin (2), xylan (1), starch (3), glucose (0.4), yeast extract (3), peptone (3), mucin type II (1), NaCl (0.9), KH2PO4 (0.45), K2HPO4 (0.45), CaCl2 (0.12), MgSO4·7H2O (0.09), hemin (0.02), and a vitamin mixture [(μg / L) biotin (10), cobalamin (10), p-aminobenzoic acid (30), folic acid (50), pyridoxamine (150), riboflavin (50), and thiamine-HCl (50)]. The digestion process was simulated by acidifying the feed to approximately pH 2 with 6M HCl, adding 10 mg / L of pepsin, and then incubating the feed at 37°C for 30–45 minutes. The feed was then neutralized to pH 6.9 with simulated pancreatic juice [(g / L)NaHCO3 (12.5), Ox bile salt (6), and pancreatin (0.9)], and 0.5 g / L of cysteine ​​was added. This feed was continuously supplied into a luminal chamber at a rate that provided a retention time of approximately 24 hours, and the pH was maintained between 6.6 and 6.9 using a pH controller and administration pump (Black Stone BL7916-2, Hanna Instruments, UK).

[0284] To initiate the SHIRM system, aliquots of fecal samples from subjects with type 2 diabetes were inoculated. A preculture was prepared in a Coy chamber (5% hydrogen, 10% carbon dioxide, and 85% nitrogen) under anaerobic conditions by adding 2% fecal material to 5 mL of LYBHI medium containing yeast extract (5 g / L), cellobiose (1 g / L), maltose (1 g / L), cysteine ​​(0.5 g / L), and hemin (0.01 g / L). The preculture was incubated at 37°C for 5 hours and then seeded into the lumen compartment of the SHIRM system. After incubating the lumen group for 24 hours, the UrdA enzyme activity test was performed.

[0285] The enzymatic reaction was carried out under anaerobic conditions in a Coy chamber at 37°C with a total volume of 1 mL. The lumen group was transferred from the SHIRM system to the Coy chamber, and 1 mL aliquots were prepared for different experimental conditions containing 50 μM or 250 μM urocanic acid and UrdA inhibitor at different concentrations (the two concentrations of urocanic acid substrate were used to illustrate the possibility of slightly different kinetic parameters for polymicrobial conditions and UrdA activity of different bacterial species). The vehicle control consisted of 2.5% DMSO. The reaction mixture was incubated for 24 hours, and samples were taken at 1 hour, 2 hours, 4 hours, 8 hours, and 24 hours to capture incubation times within the linear range of product formation. The reaction was stopped by removing the samples from the anaerobic environment, and they were stored at -20°C until further analysis. Substrate conversion rates and IC50 values ​​were determined using the same method as in Example 10.

[0286] result The results for compounds 5, 1, and 9 that were tested are shown in Table 3. [Table 9]

[0287] The tested compounds showed the potential to inhibit the enzyme activity of UrdA in multimicrobial human fecal cultures from type 2 diabetic donors. This confirmed the results observed in the single-strain lysate UrdA assay using S. oneidensis, E. lenta, A. prevoti, and B. laterosporus in Example 9.

[0288] Example 13 - In vitro ADME characteristics The degree to which a compound permeates a membrane affects absorption from the gastrointestinal tract, distribution of the compound throughout the body, metabolism, and excretion patterns. Information on these properties can be obtained by studying the apparent permeability (Papp) in human cell monolayers (Hubatsch et al., Nature Protocols, 2007).

[0289] Caco-2 cell monolayers were grown on a permeable filter support. On day 21 after seeding, the Caco-2 filter was washed with pre-warmed HBSS (pH 7.4), and a compound (UrdA inhibitor) (1 μM in HBSS) was added to either the apical or basal side. The cells were then incubated at 37°C on an orbital shaker (500 rpm). After 30 minutes, samples were removed from both compartments. The samples (100 μL) were transferred to a 96-well plate containing 100 μL of methanol and warfarin as an IS, and analyzed by LC-MS / MS. 5 μM enalaprilat was used as a membrane integrity control for each filter. The Papp of enalaprilat for dense monolayers was determined to be <1 × 10⁻⁶ cm / s.

[0290] Compounds 8, 5, 1, and 6 showed permeability through the Caco2 cell monolayer, and the efflux ratio did not increase (Table 4). The results indicate that the compounds can be absorbed from the gastrointestinal tract and enter the systemic circulation. [Table 10]

[0291] The proportion of plasma protein-bound and unbound compounds, and plasma stability were evaluated in the presence of human and mouse plasma. The fraction of unbound compounds (fu) in plasma derived from human (UU0938013) and mouse (CD1 / K2 EDTA, lot: 34617, 2021-05-06, Innovative Research) was determined by rapid equilibrium dialysis (RED) using a ThermoFisher Scientific apparatus at 37°C for 4 hours. The compound (UrdA inhibitor) was added to plasma samples at concentrations of 0.1, 1, or 10 μM and dialyzed against isotonic phosphate buffer (67 mM, pH 7.4). After dialyzing, the compound concentrations in the buffer and plasma were quantified by LC-MS / MS analysis. In parallel, the stability of the UrdA inhibitor in plasma was determined by incubating plasma containing the compound (0.1, 1, and 10 μM, respectively) at 37°C for 4 hours. The compound concentration was quantified by LC-MS / MS analysis.

[0292] The results for compounds 8, 5, 1, and 9, which were tested, are shown in Tables 5 and 6. Compounds 8, 5, 1, and 6 showed low plasma protein binding in the presence of human and mouse plasma and were stable in both human and mouse plasma. [Table 11] [Table 12]

[0293] The metabolic stability of the compound (UrdA inhibitor) was tested by measuring the in vitro half-life (t1 / 2) in human (50 pooled, mixed sex) and mouse (CD-1, male) liver microsomes. For this purpose, 0.5 mg / mL of human or mouse liver microsomes were incubated with 1 μM of the compound (UrdA inhibitor) in 100 mM KPO4 buffer (pH 7.4) at a total incubation volume of 500 μL. The reaction was initiated by adding 1 mM NADPH, and samples were collected at different incubation times up to 300 minutes. The reaction was stopped by adding cold acetonitrile, and the amount of residual parent compound was analyzed by LC-MS / MS.

[0294] The results for compounds 8, 5, 1, and 6 that were tested are shown in Tables 7 and 8. Compounds 8, 5, 1, and 6 showed very high microsomal stability in human liver microsomes and mouse liver microsomes (except for compound 5, which had a t1 / 2 of 120 minutes). [Table 13] [Table 14]

Claims

1. A compound of formula (I), its stereoisomer, tautomer, or pharmaceutically acceptable salt, for use in preventing, inhibiting, or treating conditions or disorders mediated by urocanate reductase, 【Chemistry 1】 During the ceremony, A is a five-membered heteroaromatic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, wherein at least one of the heteroatoms is nitrogen. R １ and R ２ but, -H, C １－６ Alkyl, preferably C １－３ Alkyl, for example -CH ３ , C １－６ Haloalkyl, preferably C １－３ Haloalkyl, e.g., -CF ３ , C １－６ alkoxy, preferably C １－３ alkoxy, for example -OCH ３ , Halogens, preferably F, Cl, Br, or I, and - Selected independently from CN, If present, each R ３ but, C １－６ Alkyl, preferably C １－３ Alkyl, for example -CH ３ , C １－６ Haloalkyl, preferably C １－３ Haloalkyl, e.g., -CF ３ , C １－６ Alkoxy, preferably C １－３ Alkoxy, for example, -OCH ３ , Halogen, preferably F, Cl, Br, or I, -OH, - CN, -NO ２ 、 -NR ５ R ６ (In the formula, R ５ and R ６ However, H and C １－３ A component independently selected from alkyl groups, preferably H and -CH. ３ (Selected from), and -C (=NR ７ ) (NR ８ R ９ ) (wherein, R ７ , R ８ , and R ９ Each of them is H and C １－３ A component independently selected from alkyl groups, preferably H and -CH. ３ Selected independently from (selected from), R ４ but, -OH -SH C １－６ Alkoxy, preferably C １－３ Alkoxy, and -NR １０ R １１ (In the formula, R １０ and R １１ However, H and C １－３ A component independently selected from alkyl groups, preferably H and -CH. ３ Selected from) and n is an integer between 0 and 2. Compounds of formula (I), stereoisomers thereof, tautomers thereof, or pharmaceutically acceptable salts thereof.

2. The compound for use according to claim 1, wherein the condition or disorder is a gastrointestinal disorder or condition, a cardiovascular disease or disorder, or a metabolic disorder.

3. The compound for use according to claim 1, wherein the condition or disorder is primary sclerosing cholangitis (PSC), myocardial fibrosis, heart failure, type 2 diabetes, or prediabetes.

4. The compound for use according to any one of claims 1 to 3, wherein in formula (I), A is a five-membered heteroaromatic ring containing one to three heteroatoms selected from nitrogen and sulfur, preferably a five-membered heteroaromatic ring containing one or two heteroatoms selected from nitrogen and sulfur.

5. The compound for use according to any one of claims 1 to 3, wherein in formula (I), A is a five-membered heteroaromatic ring containing one to three nitrogen atoms, preferably a five-membered heteroaromatic ring containing one or two nitrogen atoms.

6. A compound for use according to any one of claims 1 to 3, wherein in formula (I), A is imidazolyl, pyrazolyl, pyrrolyl, or thiazolyl, preferably 4-imidazolyl, 5-imidazolyl, 3-pyrazolyl, 4-pyrazolyl, 3-pyrrolyl, or 4-thiazolyl.

7. A compound for use according to any one of claims 1 to 6, wherein in formula (I), n is 0 or 1.

8. In equation (I), n is non-zero, and R ３ However, C １－３ Alkyl (e.g., -CH ３ or -CH ２ CH ３ ), C １－３ Haloalkyl (e.g., CF) ３ ), C １－３ Alkoxy (e.g., -OCH) ３ ), F, Cl, -NH ２ A compound for use according to any one of claims 1 to 6, selected from , and -OH.

9. In equation (I), R １ and R ２ However, -H, C １－３ Alkyl (e.g., -CH) ３ or -CH ２ CH ３ ), C １－３ Haloalkyl (e.g., -CF) ３ A compound for use according to any one of claims 1 to 8, independently selected from ), F, and Cl.

10. In equation (I), R １ However, H, -CH ３ ien-CH ２ CH ３ A compound for use according to any one of claims 1 to 9, wherein the compound is F, or F.

11. R １ A compound for use according to any one of claims 1 to 9, wherein H is present.

12. In equation (I), R ２ However, H, -CH ３ ien-CH ２ CH ３ , or F, preferably H, -CH ３ , or -CH ２ CH ３ A compound for use according to any one of claims 1 to 11.

13. R ２ However, H, -CH ３ , or -CH ２ CH ３ The compound for use according to claim 12.

14. R ２ However, -CH ３ or -CH ２ CH ３ Preferably -CH ３ The compound for use according to claim 13.

15. R ２ A compound for use according to any one of claims 1 to 11, wherein the compound is other than H.

16. In equation (I), R ４ However, -OH or C １－３ A compound for use according to any one of claims 1 to 15, which is an alkoxy, preferably an -OH group.

17. In formula (I), the heteroaromatic ring A and the group -C(O)R ４ A compound for use according to any one of claims 1 to 16, wherein the compounds are trans relative to each other.

18. The compound is the compound of formula (II), its stereoisomer, tautomer, or a pharmaceutically acceptable salt. 【Chemistry 2】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ is as defined in any one of claims 1, 9, and 12 to 15, R ３ However, as defined in claim 1 or claim 8, R ４ However, as defined in claim 1 or claim 16, and A compound for use according to any one of claims 1 to 3, wherein m is an integer from 0 to 2, preferably 0 or 1.

19. The compound comprises the imidazolyl ring and the group -C(O)R ４ The compound for use according to claim 18, wherein the compound is a compound of formula (II) that is trans to each other, a tautomer thereof, or a pharmaceutically acceptable salt thereof.

20. The compound is the compound of formula (III), its stereoisomer, tautomer, or a pharmaceutically acceptable salt. 【Transformation 3】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ However, as defined in any one of claims 1, 9, and 12-15, R ３ However, as defined in claim 1 or claim 8, R ４ is as defined in claim 1 or claim 16, and The compound for use according to any one of claims 1 to 3, wherein p is an integer from 0 to 2, preferably 0 or 1.

21. The compound comprises the pyrazolyl ring and the group -C(O)R ４ The compound for use according to claim 20, wherein the compound is a compound of formula (III) that is trans to each other, a tautomer thereof, or a pharmaceutically acceptable salt thereof.

22. The compound is the compound of formula (IV), its stereoisomer, tautomer, or a pharmaceutically acceptable salt. 【Chemistry 4】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ However, as defined in any one of claims 1, 9, and 12-15, R ３ This is as defined in claim 1 or claim 8, R ４ However, as defined in claim 1 or claim 16, and A compound for use according to any one of claims 1 to 3, wherein q is an integer from 0 to 2, preferably 0 or 1.

23. The compound comprises the pyrrolyl ring and the group -C(O)R ４ The compound for use according to claim 22, wherein the compound is a compound of formula (IV) that is trans to each other, a tautomer thereof, or a pharmaceutically acceptable salt thereof.

24. The compound is a compound of formula (V), its stereoisomer, tautomer, or a pharmaceutically acceptable salt. 【Transformation 5】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ However, as defined in any one of claims 1, 9, and 12-15, R ３ However, as defined in claim 1 or claim 8, R ４ However, as defined in claim 1 or claim 16, and The compound for use according to any one of claims 1 to 3, wherein s is an integer from 0 to 2, preferably 0 or 1.

25. The compound comprises the thiazolyl ring and the group -C(O)R ４ The compound for use according to claim 24, wherein the compound is a compound of formula (V) that is trans to each other, a tautomer thereof, or a pharmaceutically acceptable salt thereof.

26. The compound of formula (I) is as follows: Table 1-1 Table 1-2 Table 1-3 A compound for use according to any one of claims 1 to 3, which is a compound selected from, stereoisomers thereof, tautomers thereof, and pharmaceutically acceptable salts thereof.

27. The compound for use according to claim 26, wherein the compound is selected from compound numbers 1 to 28, their tautomers, and their pharmaceutically acceptable salts, wherein the compound is selected from compound numbers 1 to 28, in which the five-membered heteroaromatic ring is trans with respect to the -C(O)OH group.

28. A compound of formula (II), its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Transformation 6】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ However, as defined in any one of claims 1, 9, and 12-15, R ３ However, as defined in claim 1 or claim 8, R ４ However, as defined in claim 1 or claim 16, and m is an integer between 0 and 2, preferably 0 or 1. However, if the compound is 【Transformation 7】 That is unexpected. Compounds of formula (II), stereoisomers thereof, tautomers thereof, or pharmaceutically acceptable salts thereof.

29. In equation (II), either m is 0 or m is 1 and R ３ However, base-NR ５ R ６ And preferably -NH ２ The compound according to claim 28.

30. In equation (II), R １ and R ２ However, -H and C １－６ A component independently selected from alkyl groups, preferably -H and C. １－３ Selected independently of alkyl, for example, -H and -CH ３ A compound according to claim 28 or 29, independently selected from the above.

31. In equation (II), R ４ The compound according to any one of claims 28 to 30, wherein the compound is an OH group.

32. A compound of formula (IIa), its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Transformation 8】 In the formula, R ３ The compound according to claim 28, wherein m is as defined in claim 28.

33. In equation (IIa), either m is 0 or m is 1 and R ３ However, base-NR ５ R ６ or preferably -NH ２ is or R ３ C １－３ It is alkyl, or preferably -CH ３ The compound according to claim 32.

34. imidazolyl ring and the group -C(O)R ４ The compound according to any one of claims 28 to 33, wherein the compound is a compound of formula (II) in trans relative to each other, or a compound of formula (IIa) in trans relative to each other, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

35. A compound of formula (III), or its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Chemistry 9】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ However, as defined in any one of claims 1, 9, and 12-15, R ３ However, as defined in claim 1 or claim 8, R ４ However, as defined in claim 1 or claim 16, and p is an integer between 0 and 2, preferably 0 or 1. However, if the compound is 【Chemistry 10】 That is unexpected. The compound of formula (III), or its stereoisomers, tautomers, or pharmaceutically acceptable salts.

36. In equation (III), p is either 0 or p is 1, R ３ C １－６ It is alkyl, preferably C １－３ It is alkyl, for example -CH ３ The compound according to claim 35.

37. In equation (III), R １ and R ２ However, -H and C １－６ A component independently selected from alkyl groups, preferably -H and C. １－３ Selected independently of alkyl groups, e.g., -H, -CH ３ and -CH ２ CH ３ A compound according to claim 35 or 36, independently selected from the above.

38. In equation (III), R ４ The compound according to any one of claims 35 to 37, wherein the compound is an OH group.

39. The compound of formula (IIIa), its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Chemistry 11】 In the formula, R ３ The compound according to claim 35, wherein p is as defined in claim 35.

40. In equation (IIIa), p is either 0 or p is 1, R ３ C １－３ It is alkyl, preferably -CH ３ The compound according to claim 39.

41. The pyrazolyl ring and the group -C(O)R ４ The compound according to any one of claims 35 to 40, wherein the compound is a compound of formula (III) in trans relative to each other, or a compound of formula (IIIa) in trans relative to each other, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

42. A compound of formula (IV), its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Chemistry 12】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ However, as defined in any one of claims 1, 9, and 12-15, R ３ However, as defined in claim 1 or claim 8, R ４ However, as defined in claim 1 or claim 16, and q is an integer between 0 and 2, preferably 0 or 1. However, if the compound is 【Chemistry 13】 That is unexpected. Compounds of formula (IV), stereoisomers thereof, tautomers thereof, or pharmaceutically acceptable salts thereof.

43. The compound according to claim 42, wherein q is 0 in formula (IV).

44. In equation (IV), R １ and R ２ However, -H and C １－６ A component independently selected from alkyl groups, preferably -H and C. １－３ Selected independently of alkyl, for example, -H and -CH ３ A compound according to claim 42 or 43, independently selected from the above.

45. In equation (IV), R ４ The compound according to any one of claims 42 to 44, wherein the compound is an OH group.

46. A compound of formula (IVa), its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Chemistry 14】 In the formula, R ３ The compound according to claim 42, wherein and q are as defined in claim 42.

47. The pyrrolyl ring and the group -C(O)R ４ The compound according to any one of claims 42 to 46, wherein the compound is a compound of formula (IV) in trans relative to each other, or a compound of formula (IVa) in trans relative to each other, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

48. A compound of formula (V), or its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Chemistry 15】 During the ceremony, R １ However, as defined in any one of claims 1 and 9 to 11, R ２ However, as defined in any one of claims 1, 9, and 12-15, R ３ However, as defined in claim 1 or claim 8, R ４ However, as defined in claim 1 or claim 16, and s is an integer between 0 and 2, preferably 0 or 1. However, if the compound is 【Chemistry 16】 That is unexpected. A compound of formula (V), or its stereoisomers, tautomers, or pharmaceutically acceptable salts.

49. In equation (V), s is 1, and R ３ However, base-NR ５ R ６ And preferably -NH ２ The compound according to claim 48.

50. In equation (V), R １ and R ２ The compound according to claim 48 or 49, wherein both are -H.

51. In equation (V), R ４ The compound according to any one of claims 48 to 50, wherein the compound is an OH group.

52. A compound of formula (Va), its stereoisomer, tautomer, or pharmaceutically acceptable salt, 【Chemistry 17】 In the formula, R ３ The compound according to claim 48, wherein and s are as defined in claim 49.

53. The thiazolyl ring and the group -C(O)R ４ The compound according to any one of claims 48 to 52, wherein the compound is a compound of formula (V) in trans relative to each other, or a compound of formula (Va) in trans relative to each other, or a tautomer thereof, or a pharmaceutically acceptable salt thereof.

54. The compound has the following structure: Table 2-1 Table 2-2 The compound according to any one of claims 28 to 53, or having a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

55. The compound according to claim 54, wherein the compound is selected from compound numbers 5, 8, 15, 16, 17, 18, 20, 21, 25, 28, 1, 4, 6, 25, 27, 22, 23, and 2, their tautomers, and their pharmaceutically acceptable salts.

56. A compound according to any one of claims 28 to 55, for use as a pharmaceutical.

57. A pharmaceutical composition comprising a compound according to any one of claims 28 to 55, together with one or more pharmaceutically acceptable carriers, excipients, or diluents.

58. Use of the compound according to any one of claims 1 and 4 to 27 in the manufacture of a pharmaceutical product for use in preventing, inhibiting or treating a condition or disorder mediated by urocanate reductase.

59. The use according to claim 58, wherein the condition or disorder is a gastrointestinal disorder or condition, a cardiovascular disease or disorder, or a metabolic disorder.

60. The use according to claim 58, wherein the condition or disorder is primary sclerosing cholangitis (PSC), myocardial fibrosis, heart failure, type 2 diabetes, or prediabetes.

61. A method for preventing, inhibiting, or treating a condition or disorder mediated by urocanate reductase, comprising the step of administering a pharmaceutically effective amount of any one of claims 1 and 4 to 27 to a patient (e.g., a human subject) in need thereof.

62. The method according to claim 61, wherein the condition or disorder is a gastrointestinal disorder or condition, a cardiovascular disease or disorder, or a metabolic disorder.

63. The method according to claim 61, wherein the condition or disorder is primary sclerosing cholangitis (PSC), myocardial fibrosis, heart failure, type 2 diabetes, or prediabetes.

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