Compounds, acetaldehyde dehydrogenase 2 activators, pharmaceutical compositions, and therapeutic and / or preventative agents.

Abstract

This invention provides compounds with acetaldehyde dehydrogenase 2 (ALDH2) activating activity, etc. The following formula (1) [where A is a heterocycle, R...] 1 and R 2 Each can be independently hydrogen, alkyl, alkenyl, or alkynyl, R 3 X is alkyl, alkenyl, or alkynyl. 1 and X 2 Each of these refers independently to a compound represented by a halogen, its pharmaceutically permissible salt, or its prodrug.

Description

Technical Field

[0001] This invention relates to compounds, acetaldehyde dehydrogenase 2 activators, pharmaceutical compositions, and therapeutic and / or preventative medicines. Background Technology

[0002] Aldehyde dehydrogenase 2 (ALDH2) is an enzyme that breaks down aldehydes such as acetaldehyde. Its relationship with various diseases has been reported, such as Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain.

[0003] Fanconi anemia (FA) is a hereditary bone marrow failure disorder accompanied by symptoms such as aplastic anemia, leukemia, cancer, and malformations. Properly breaking down aldehydes and repairing damaged genomes is important in the blood-producing stem cells in the bone marrow; however, there are reports of FA patients being unable to repair genoscopic damage caused by aldehydes, leading to the development of anemia (e.g., Non-Patent Literature 1).

[0004] Regarding its relationship with osteoporosis, reports indicate that osteoporosis symptoms and decreased bone density were observed in ALDH2 gene mutation model mice; the osteoblast differentiation capacity of the aforementioned model mice was significantly reduced; and in humans, the osteoblast formation capacity of ALDH2 gene mutations was also reduced (e.g., Non-Patent Literature 2).

[0005] NAFLD and NASH are metabolic diseases caused by the accumulation of triglycerides in the liver. It has been reported that the incidence of NAFLD is high in individuals with low ALDH2 activity genotypes (e.g., Non-Patent Literature 3). Furthermore, alcoholic liver injury and pancreatitis are caused by excessive alcohol consumption, with acetaldehyde, produced from the breakdown of ethanol in the body, considered the primary cause. In addition, it has been reported that ALDH2 activating compounds or the introduction of the ALDH2 gene have improved symptoms in pathological models of NAFLD, NASH, alcoholic liver injury, and pancreatitis (e.g., Non-Patent Literature 4, 5, and 6).

[0006] Ischemia-reperfusion injury is tissue damage that occurs when blood supply to an organ is restored after prolonged ischemia due to arterial obstruction. ALDH2-activating compounds have been reported to be effective in protecting against ischemia-reperfusion injury (e.g., Patent Document 1). Furthermore, an association between peripheral artery disease and ALDH2 has been suggested (e.g., Non-Patent Document 7).

[0007] Alzheimer's disease and Parkinson's disease are neurodegenerative diseases of unknown cause, but there are reports that alcohol consumption and ALDH2 gene mutations contribute to the onset and progression of the symptoms (e.g., non-patent literature 8 and 9).

[0008] Risk factors for esophageal cancer are considered to be smoking, alcohol consumption, and excessive intake of hot drinks and foods. The International Agency for Research on Cancer (IARC) classified alcoholic beverages as carcinogenic to esophageal squamous cell carcinoma in 2010. The association between esophageal cancer and alcohol consumption and ALDH2 has been reported; for example, analyses of esophageal and head and neck cancer patients suggest the possibility of inhibiting esophageal cancer progression through alcohol abstinence; in mice with a knock-in low-activity mutant of the human ALD H2 gene, alcohol consumption resulted in more intense DNA damage in the esophagus; and so on (e.g., Non-Patent Literature 10 and 11). Similarly, the association between head and neck cancer and ALDH2 has been reported (e.g., Non-Patent Literature 12).

[0009] Pain is generally known to include inflammatory (damage-sensitizing), neuropathic, and unexplained pain. It has been reported that in mouse models of inflammatory pain, mice with ALDH2 mutants are more likely to experience painful stimuli, which can be relieved by ALDH2-activating compounds (e.g., Non-Patent Literature 13).

[0010] As mentioned above, the relationship between ALDH2 and various diseases has been reported, and therefore it is expected that ALDH2 activation will be effective for the treatment and / or prevention of various diseases. Compounds with ALD H2 activating activity are known, for example, those described in Patent Documents 1-4.

[0011] Existing technical documents

[0012] Patent documents

[0013] Patent Document 1: International Publication No. 2008 / 112164

[0014] Patent Document 2: International Publication No. 2014 / 160185

[0015] Patent Document 3: International Publication No. 2015 / 127137

[0016] Patent Document 4: International Publication No. 2019 / 151241

[0017] Non-patent literature

[0018] Non-patent literature 1: Blood (2013) 122(18): 3206-3209

[0019] Non-patent literature 2: Journal of Bone and Mineral Research (2012) 27(9): 2015-2023

[0020] Non-patent literature 3: Nutrition & Diabetes (2016) 6, e210

[0021] Non-patent literature 4: Redox Biology (2019) 24:101205

[0022] Non-patent literature 5: Journal of Hepatology (2015) 62: 647-656

[0023] Non-patent literature 6: Biochemical and Biophysical Research Communications (2020) 522:518-524

[0024] Non-patent literature 7: Pharmacological Research (2017) 115: 96-106

[0025] Non-patent literature 8: Biochemical and Biophysical Research Communications (2000) 273: 192-196

[0026] Non-patent literature 9: Neurology (2014) 82: 419-426

[0027] Non-patent literature 10: Gastroenterology (2016) 151:860-869

[0028] Non-patent literature 11: Carcinogenesis (2020) 41:194-202

[0029] Non-patent literature 12: International Journal of Oncology (2008) 32: 945-973

[0030] Non-patent literature 13: Science Translational Medicine (2014) 6:251ra118 Summary of the Invention

[0031] The problem that the invention aims to solve

[0032] The purpose of this invention is to provide compounds having ALDH2 activating activity, or ALDH2 activators containing the aforementioned compounds, pharmaceutical compositions, or therapeutic or preventive agents.

[0033] Methods for solving problems

[0034] The inventors of this application conducted in-depth research and discovered that compounds with a specified structure have ALDH2 activating effects, thus completing this invention.

[0035] The present invention includes the following embodiments. [1]

[0037] The compounds represented by the following formula (1), their pharmaceutically permissible salts, or their prodrugs.

[0038] [Chemical Formula 1]

[0039]

[0040] [In the formula,

[0041] A is a heterocyclic ring.

[0042] R 1 and R 2 Each can be independently hydrogen, alkyl, alkenyl, or alkynyl.

[0043] R 3 It can be alkyl, alkenyl, or ynyl.

[0044] X 1 and X 2 Each is an independent halogen. [2]

[0046] Compounds, pharmaceutically permissible salts thereof, or their prodrugs as described in [1], wherein A contains at least one nitrogen atom as a ring member atom. [3]

[0048] Compounds as described in [1] or [2], their pharmaceutically permissible salts or their prodrugs, wherein A is a 5- or 6-membered ring. [4]

[0050] Compounds, pharmaceutically permissible salts or prodrugs thereof, as described in any one of [1] to [3], wherein A is an aromatic heterocycle. [5]

[0052] The compound, a pharmaceutically permissible salt thereof, or a prodrug thereof, as described in any one of [1] to [4], wherein the compound represented by formula (1) is the compound represented by formula (2) or (3) below.

[0053] [Chemical Formula 2]

[0054]

[0055] [Chemical Formula 3]

[0056]

[0057] [In the formula, R] 1 R 2 R 3 X 1 and X 2 As mentioned above] [6]

[0059] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [5], wherein R 1 It is hydrogen or alkyl.

[0060] [6-1]

[0061] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [6], wherein R 1 It can be hydrogen or methyl.

[0062] [6-2]

[0063] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [6-1], wherein R 1 It is hydrogen. [7]

[0065] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [6-2], wherein R 2 It is hydrogen or alkyl.

[0066] [7-1]

[0067] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [7], wherein R 2 It can be hydrogen or methyl.

[0068] [7-2]

[0069] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [7-1], wherein R 2 It is hydrogen. [8]

[0071] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [7-2], wherein R 3 It is an alkyl group that has been substituted with a halogen or an unsubstituted alkyl group.

[0072] [8-1]

[0073] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [8], wherein R 3 It can be a fluorinated methyl group or an unsubstituted methyl group.

[0074] [8-2]

[0075] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [8-1], wherein R 3 The methyl group is unsubstituted. [9]

[0077] Compounds, pharmaceutically permissible salts thereof, or their prodrugs as described in any one of [1] to [8-2], wherein X 1 It is either fluorine or chlorine.

[10]

[0079] Compounds, pharmaceutically permissible salts thereof, or prodrugs thereof, as described in any one of [1] to [9], wherein X 2 It is fluorine.

[11]

[0081] The compound, a pharmaceutically permissible salt thereof, or a prodrug thereof, as described in any one of [1] to

[10] , wherein the compound represented by formula (1) is selected from the group consisting of the following compounds.

[0082] [Chemical Formula 4]

[0083]

[12]

[0085] The prodrug or a pharmaceutically permissible salt thereof as described in any one of [1] to

[11] , wherein R 1 It is -CH2-O-PO3H2.

[13]

[0087] The prodrug or a pharmaceutically permissible salt thereof as described in any one of [1] to

[11] , wherein A contains at least one nitrogen atom as a ring member atom, at least one of the aforementioned nitrogen atoms being replaced by -CH2-O-PO3H2.

[14]

[0089] The prodrug or its pharmaceutically permissible salt as described in any one of [1] to

[13] , wherein the prodrug or its pharmaceutically permissible salt is selected from the group consisting of the following compounds.

[0090] [Chemical Formula 5]

[0091]

[15]

[0093] Acetaldehyde dehydrogenase 2 activator, comprising any one of the compounds described in [1] to

[14] , a pharmaceutically permissible salt thereof, or a prodrug thereof.

[16]

[0095] A pharmaceutical composition comprising any one of the compounds described in [1] to

[14] , a pharmaceutically permissible salt thereof, or a prodrug thereof.

[17]

[0097] The treatment and / or prevention of diseases selected from the group consisting of Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain, wherein the treatment and / or prevention comprises any one of [1] to

[14] , a pharmaceutically permissible salt thereof, or a prodrug thereof.

[0098] [A1]

[0099] A method for activating acetaldehyde dehydrogenase 2, the method comprising administering to a patient in need an effective amount of any one of [1] to

[14] , a pharmaceutically permissible salt thereof, or a prodrug thereof.

[0100] [A2]

[0101] Methods for treating and / or preventing diseases selected from the group consisting of Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain, wherein the methods comprise administering to a patient in need an effective amount of any one of [1] to

[14] compounds, their pharmaceutically permissible salts, or their prodrugs.

[0102] [B1]

[0103] The compound, pharmaceutically permissible salt thereof, or prodrug thereof, as described in any one of [1] to

[14] , is used for the activation of acetaldehyde dehydrogenase 2.

[0104] [B2]

[0105] The compound, a pharmaceutically permissible salt thereof, or a prodrug thereof, as described in any one of [1] to

[14] , is used for the treatment and / or prevention of a disease selected from the group consisting of Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain.

[0106] [C1]

[0107] The use of any compound, pharmaceutically permissible salt thereof, or prodrug thereof as described in any one of [1] to

[14] for the activation of acetaldehyde dehydrogenase 2.

[0108] [C2]

[0109] The use of any compound, a pharmaceutically permissible salt thereof, or a prodrug thereof in any of [1] to

[14] for the treatment and / or prevention of diseases selected from the group consisting of Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain.

[0110] [D1]

[0111] Use of any compound, pharmaceutically permissible salt thereof, or prodrug thereof in any of [1] to

[14] in the manufacture of acetaldehyde dehydrogenase 2 activator.

[0112] [D2]

[0113] The use of any compound, pharmaceutically permissible salt thereof, or prodrug thereof in the manufacture of a treatment and / or preventive medicine for a disease selected from the group consisting of Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain.

[0114] Invention Effects

[0115] According to the present invention, compounds having ALDH2 activating activity, or ALDH2 activators containing the aforementioned compounds, pharmaceutical compositions, or therapeutic or preventive agents may be provided. Detailed Implementation

[0116] The embodiments of the present invention will be described in detail below, but the present invention is not limited thereto and various modifications can be made without departing from its spirit.

[0117] <Compound>

[0118] One embodiment of the present invention relates to compounds represented by the following formula (1), their pharmaceutically permissible salts, or their prodrugs.

[0119] [Chemical Formula 6]

[0120]

[0121] [In the formula,

[0122] A is a heterocyclic ring.

[0123] R 1 and R 2 Each can be independently hydrogen, alkyl, alkenyl, or alkynyl.

[0124] R 3 It can be alkyl, alkenyl, or ynyl.

[0125] X 1 and X 2 Each is an independent halogen.

[0126] In formula (1), A preferably contains at least one nitrogen atom as a ring member atom. A is preferably a 5- or 6-membered ring. A is preferably an aromatic heterocycle. Although not particularly limited, A is preferably a pyridine ring or a pyrazole ring.

[0127] In equation (1), R 1 ~R 3 The alkyl, alkenyl, and alkynyl groups can be linear or branched. R 1 ~R 3 The alkyl, alkenyl, and alkynyl groups can be substituted or left unsubstituted. Examples of such substituents include halogens (fluorine, chlorine, bromine, or iodine). The number of substituents can be, for example, one, two, or three.

[0128] In equation (1), R 1 ~R 3 Each alkyl group is preferably an alkyl group having 1 to 6 carbon atoms, more preferably an alkyl group having 1 to 3 carbon atoms, and even more preferably a methyl group. 1 ~R 3 Each alkenyl group is preferably an alkenyl group having 2 to 6 carbon atoms, and more preferably an alkenyl group having 2 to 4 carbon atoms. R 1 ~R 3 Each alkynyl group is preferably an alkynyl group with 2 to 6 carbon atoms, and more preferably an alkynyl group with 2 to 4 carbon atoms.

[0129] In equation (1), considering the activity and metabolic stability of ALDH2, R 1 Preferably hydrogen or alkyl, more preferably hydrogen or methyl, and even more preferably hydrogen.

[0130] In equation (1), considering the activity and metabolic stability of ALDH2, R 2 Preferably hydrogen or alkyl, more preferably hydrogen or methyl, and even more preferably hydrogen.

[0131] In equation (1), from the perspective of metabolic stability, R 3 Preferably, it is a halogen-substituted alkyl group or an unsubstituted alkyl group, more preferably a fluorine-substituted methyl group or an unsubstituted methyl group, and even more preferably an unsubstituted methyl group.

[0132] In equation (1), X 1 The preferred form is fluorine, chlorine, bromine, or iodine, more preferably fluorine or chlorine, and even more preferably fluorine.

[0133] In equation (1), X 2 The preferred form is fluorine, chlorine, bromine, or iodine, more preferably fluorine or chlorine, and even more preferably fluorine.

[0134] The compound represented by formula (1) is not particularly limited, but is preferably represented by the compound represented by formula (2) or (3) below.

[0135] [Chemical Formula 7]

[0136]

[0137] [Chemical Formula 8]

[0138]

[0139] [In the formula, R] 1 R 2 R 3 X 1 and X 2 As mentioned above]

[0140] The compound represented by formula (1) is not particularly limited, but is preferably the following compound.

[0141] [Chemical Formula 9]

[0142]

[0143] As prodrugs for the aforementioned compounds, examples include phosphorylated prodrugs, and more specifically, R... 1The prodrug is -CH2-O-PO3H2, and at least one of the nitrogen atoms that are ring member atoms of A is replaced by -CH2-O-PO3H2.

[0144] There are no particular limitations on the prodrug or its pharmaceutically permissible salt, but the following compounds are preferred.

[0145] [Chemical Formula 10]

[0146]

[0147] The pharmaceutically permissible salts of the aforementioned compounds or prodrugs are not particularly limited as long as they can be used in medicine. Examples include inorganic acid salts such as hydrochloride, sulfate, nitrate, phosphate, and hydrobromide; organic acid salts such as fumarate, maleate, malate, tartrate, succinate, citrate, methanesulfonate, p-toluenesulfonate, acetate, lactate, and palmitate; alkali metal salts; and alkaline earth metal salts.

[0148] The aforementioned compounds, their pharmaceutically permissible salts, or their prodrugs can also form solvates such as hydrates. In this specification, solvates are included in the aforementioned compounds, their pharmaceutically permissible salts, or their prodrugs.

[0149] In cases where the aforementioned compounds, their pharmaceutically permissible salts, or their prodrugs exist in stereoisomers (e.g., enantiomers, diastereomers), the individual stereoisomers and mixtures thereof (e.g., racemates) are included in the aforementioned compounds, their pharmaceutically permissible salts, or their prodrugs.

[0150] <Acetaldehyde dehydrogenase 2 activator>

[0151] One embodiment of the present invention relates to an ALDH2 activator comprising the above-described compounds, their pharmaceutically permissible salts, or their prodrugs. The ALDH2 activator of this embodiment not only exhibits excellent ALDH2 activation but also possesses superior properties (e.g., excellent metabolic stability, inhibition of reactive metabolite formation, and avoidance of hERG inhibition).

[0152] By using the ALDH2 activator of this embodiment, it is possible to treat and / or prevent ALDH2-related diseases.

[0153] <Pharmaceutical compositions and therapeutic and / or preventive agents>

[0154] One embodiment of the present invention relates to a pharmaceutical composition comprising the above-described compounds, pharmaceutically permissible salts thereof, or prodrugs thereof. Another embodiment of the present invention relates to a therapeutic and / or preventative medicament comprising the above-described compounds, pharmaceutically permissible salts thereof, or prodrugs thereof.

[0155] Diseases that may be targeted by the pharmaceutical composition and treatment and / or prevention drugs of this embodiment include, for example, Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain.

[0156] The pharmaceutical compositions and therapeutic and / or preventive agents of this embodiment can be administered orally or non-orally. Examples of dosage forms for oral administration include tablets, pills, granules, powders, capsules, syrups, emulsions, and suspensions. Examples of dosage forms for non-oral administration include injections, infusions, drops, eye drops, and suppositories.

[0157] The pharmaceutical compositions and therapeutic and / or preventive agents of this embodiment may include, as needed, excipients, binders, lubricants, disintegrants, sweeteners, surfactants, suspending agents, emulsifiers, colorants, preservatives, fragrances, flavoring agents, stabilizers, thickeners, etc.

[0158] The dosage of the pharmaceutical composition and the therapeutic and / or preventive drugs in this embodiment (based on the active ingredient) varies depending on the patient's condition, weight, type of compound, type of disease, route of administration, etc., and the physician can determine the appropriate dosage. As an example, in the treatment of Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain, for adults (weighing approximately 60 kg), 1 to 2000 mg can be administered orally, and 0.01 to 200 mg can be administered non-orally.

[0159] <Methods for manufacturing compounds>

[0160] The aforementioned compounds or their pharmaceutically permissible salts can be suitably synthesized using known methods. As an example of a synthetic method, synthetic route A can be described below.

[0161] [Chemical Formula 11]

[0162] <Synthesis Path A>

[0163]

[0164] [R 2 R 3 X 1 and X 2As mentioned above, X A and X B Each is an independent halogen.

[0165] In step A1, compound (A1) is esterified using iodomethane to obtain compound (A2).

[0166] In step A2, compound (A2) is reacted with a reducing agent (e.g., sodium borohydride) to obtain compound (A3).

[0167] In step A3, after reacting compound (A3) with a strong base (e.g., sodium hydride), it is then reacted with R. 3 The reaction of halides (e.g., iodomethane) yields compound (A4).

[0168] In step A4, compound (A4) is cyano-treated with a cyaniding agent (e.g., zinc cyanide) to obtain compound (A5).

[0169] In step A5, compound (A5) is reacted with a reducing agent (e.g., a boronane dimethyl sulfide complex) to obtain compound (A6).

[0170] In step A6, compound (A6) reacts with compound (A7) to obtain compound (A8).

[0171] In step A7, compound (A8) is reacted with bis(pinacol)diboron to obtain compound (A9).

[0172] In step A8, compound (A9) is reacted with 2-halopyridine-3-carboxaldehyde to give compound (A10).

[0173] In step A9, compound (A10) is reacted with a reducing agent (e.g., sodium borohydride) or a substance providing R. 2 The Grignard reagent reaction yields compound (A11).

[0174] Other synthetic methods for the above-mentioned compounds or their pharmaceutically permissible salts may include the following synthetic route B.

[0175] [Chemical Formula 12]

[0176] <Synthesis Path B>

[0177]

[0178] [R 2 R 3 X 1 and X 2 As mentioned above, X C It is a halogen.

[0179] In step B1, compound (B1) reacts with compound (B2) to obtain compound (B3).

[0180] In step B2, compound (B3) reacts with compound (A6) to obtain compound (B4).

[0181] In step B3, compound (B4) is reacted with a reducing agent (e.g., sodium borohydride) to obtain compound (B5).

[0182] Other synthetic methods for the above-mentioned compounds or their pharmaceutically permissible salts may include the following synthetic route C.

[0183] [Chemical Formula 13]

[0184] <Synthesis Path C>

[0185]

[0186] [R 1 ~R 3 X 1 and X 2 As mentioned above, X D It is a halogen.

[0187] In step C1, compound (C1) reacts with compound (A9) to obtain compound (C2).

[0188] The methods for synthesizing the above-mentioned compounds or their pharmaceutically permissible salts are not limited to the above-described synthetic routes A to C. Those skilled in the art can appropriately set suitable synthetic routes and reaction conditions based on the structure of the final compound. For example, if the final compound has a heterocycle other than a pyridine ring (e.g., a pyrazole ring), a compound with the corresponding heterocycle can be used instead of the compound (C1) in synthetic route C.

[0189] For the above-mentioned compounds or their pharmaceutically permissible salts as prodrugs, they can be synthesized by introducing, for example, a phosphate group using known methods.

[0190] When the compound represented by formula (1), its pharmaceutically permissible salt, or its prodrug exists in stereoisomers, the isomers can be separated using known methods. Examples of known methods include chromatography, enzymatic methods, and crystallization.

[0191] Example

[0192] The present invention will be described in more detail below using examples and comparative examples, but the technical scope of the present invention is not limited thereto.

[0193] [Manufacturing Example 1-1]

[0194] methyl 4-bromo-2-fluorobenzoate

[0195] [Chemical Formula 14]

[0196]

[0197] Iodomethane (16 mL, 0.27 mol) was added to a mixture of 4-bromo-2-fluorobenzoic acid (30 g, 0.14 mol), potassium carbonate (38 g, 0.27 mol), and N,N-dimethylformamide (DMF) (300 mL) at room temperature, and the mixture was stirred at the same temperature for 16 hours. Water (200 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (300 mL). The organic layer was dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure to give the title compound (30 g).

[0198] 1 1H-NMR spectrum (CDCl3) δ (ppm): 3.94 (3H, s), 7.81 (1H, s, J = 16 Hz), 7.32 (2H, m).

[0199] [Manufacturing Examples 1-2]

[0200] (4-Bromo-2-fluorophenyl)methanol

[0201] [Chemical Formula 15]

[0202]

[0203] Sodium borohydride (0.38 g, 10 mmol) was added to a mixture of methyl 4-bromo-2-fluorobenzoate (30 g, 0.13 mol) and methanol (300 mL) obtained in Preparation Example 1-1 at 0 °C for 30 minutes, and the mixture was stirred at 60 °C for 16 hours. Water (20 mL) was added to the reaction mixture, and the methanol was removed by distillation under reduced pressure. Water (250 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (200 mL, twice). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure to give the title compound (25 g).

[0204] 1 1H-NMR spectrum (CDCl3) δ (ppm): 4.72 (2H, s), 7.25 (1H, m), 7.34 (2H, m).

[0205] [Manufacturing Examples 1-3]

[0206] 4-Bromo-2-fluoro-1-(methoxymethyl)benzene

[0207] [Chemical Formula 16]

[0208]

[0209] At 0°C, a mixture of (25 g, 0.12 mol) methanol and DMF (50 mL) obtained in Preparation Examples 1-2 was added to a mixture of 60% sodium hydride (7.3 g, 0.18 mol) and DMF (150 mL) over 20 minutes. Iodomethane (15 mL, 0.24 mol) was added to the reaction mixture at room temperature, and the mixture was stirred at the same temperature for 16 hours. The reaction mixture was cooled to 0°C, and ice-cold water (500 mL) was added. Extraction was performed with ethyl acetate (250 mL, followed by 150 mL). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure to give the title compound (20 g).

[0210] 1 1H-NMR spectrum (CDCl3) δ (ppm): 3.40 (3H, s), 4.47 (2H, s), 7.23 (1H, m), 7.28 (2H, m).

[0211] [Manufacturing Examples 1-4]

[0212] 3-Fluoro-4-(methoxymethyl)benzonitrile

[0213] [Chemical Formula 17]

[0214]

[0215] The mixture of 4-bromo-2-fluoro-1-(methoxymethyl)benzene (20 g, 91 mmol), zinc powder (0.29 g, 4.6 mmol), zinc cyanide (16 g, 0.14 mol), and DMF (250 mL) obtained in Preparation Examples 1-3 was degassed using argon. Tris(dibenzylacetone)dipalladium(O) (4.2 g, 4.6 mmol) and 1,1'-bis(diphenylphosphine)ferrocene (5.1 g, 9.1 mmol) were added to the reaction mixture at the same temperature, and degassed again using argon. The reaction mixture was stirred at 100 °C for 16 hours. Water (500 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (500 mL, followed by 200 mL). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (15-20% ethyl acetate-n-hexane solution) to obtain the title compound (10 g).

[0216] 11H-NMR spectrum (CDCl3) δ (ppm): 3.45 (3H,s), 4.56 (2H,s), 7.35 (1H,d,J=9.2Hz), 7.48 (1H,d,J=8Hz), 7.58 (1H,t,J=14Hz).

[0217] [Manufacturing Examples 1-5]

[0218] (3-Fluoro-4-(methoxymethyl)phenyl)methylamine hydrochloride

[0219] [Chemical Formula 18]

[0220]

[0221] At 0°C, 18 mL (0.24 mol) of the borane-dimethyl sulfide complex was added dropwise over 20 minutes to a mixture of 10 g (61 mmol) of 3-fluoro-4-(methoxymethyl)benzonitrile (prepared in Examples 1-4) and 100 mL of tetrahydrofuran (THF), and the mixture was stirred at 80°C for 16 hours. The reaction mixture was cooled to 0°C, and methanol (25–30 mL) was added at the same temperature. The solvent was removed by distillation under reduced pressure. Dichloromethane (100 mL) was added to the residue, along with a 4 M solution of 1,4-dioxane hydrochloride sufficient to form a salt. The mixture was stirred at room temperature for 30 minutes. The solvent was removed by distillation under reduced pressure, and the residue was washed with diethyl ether to give the title compound (5.0 g).

[0222] 1 1H-NMR spectrum (DMSO-d6) δ (ppm): 3.37 (3H,s), 4.00 (2H,s), 4.54 (2H,s), 7.31 (1H,d,J=7.6Hz), 7.39 (1H,d,J=11Hz), 7.46 (1H,t,J=7.6Hz), 8.19 (3H,s).

[0223] [Manufacturing Examples 1-6]

[0224] 5-Bromo-2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)benzamide

[0225] [Chemical Formula 19]

[0226]

[0227] At 0 °C, diisopropylethylamine (13 mL, 46 mmol) and propane phosphoric anhydride (15 mL, 46 mmol) were added sequentially to a mixture of 5-bromo-2-fluorobenzoic acid (5.0 g, 23 mmol), (3-fluoro-4-(methoxymethyl)phenyl)methylamine hydrochloride obtained in Preparation Examples 1-5 (5.6 g, 27 mmol), and DMF (50 mL). The reaction mixture was stirred at room temperature for 3 hours. Water (200 mL) was added to the reaction mixture, and the mixture was stirred for 30 minutes. The resulting solid was filtered off and washed with water (100 mL, twice). The solid was dried under reduced pressure to give the title compound (5.2 g).

[0228] 1 1H-NMR spectrum (DMSO-d6) δ (ppm): 3.28 (3H, s), 4.46 (4H, m), 7.16 (2H, m), 7.40 (2H, m), 7.78 (2H, m), 9.03 (1H, m)

[0229] [Manufacturing Examples 1-7]

[0230] 2-Fluoro-N-(3-Fluoro-4-(methoxymethyl)benzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)benzamide

[0231] [Chemical Formula 20]

[0232]

[0233] The mixture of 5-bromo-2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)benzamide (4.0 g, 19 mmol), bis(pinacolyl)diboron (6.8 g, 10 mmol), potassium acetate (3.5 g, 9.1 mmol), and DMF (50 mL) obtained in Examples 1-6 was degassed using argon. At the same temperature, (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (0.16 g, 0.23 mmol) was added to the reaction mixture, and degassed again using argon. The reaction mixture was stirred at 100 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate (100 mL). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (15–20% ethyl acetate-n-hexane solution) to give the title compound (4 g) as crude. It was used in subsequent reactions without further purification.

[0234] [Manufacturing Examples 1-8]

[0235] 2-Fluoro-N-(3-Fluoro-4-(methoxymethyl)benzyl)-5-(3-formylpyridin-2-yl)benzamide

[0236] [Chemical Formula 21]

[0237]

[0238] Nitrogen gas was used to degas and replace the mixture of 2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)benzamide (2.0 g), 2-bromopyridine-3-carboxaldehyde (0.98 g, 4.7 mmol), potassium carbonate (1.3 g, 9.5 mmol), 1,4-dioxacyclohexane (20 mL), and water (2 mL). At the same temperature, (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (0.39 g, 0.25 mmol) was added to the reaction mixture, and the mixture was stirred at 110 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was brought to room temperature, and ethyl acetate (50 mL) was added. The mixture was filtered through diatomaceous earth. The diatomaceous earth was washed with ethyl acetate (20 mL). The solvent in the filtrate was removed by distillation under reduced pressure, and the residue was purified by silica gel column chromatography (15-20% ethyl acetate-n-hexane solution) to give the title compound (1.1 g).

[0239] 1 H-NMR spectrum (DMSO-d6) δ (ppm): 3.28 (3H, s), 4.42 (2H, s), 4.50 (2H, s), 7.18 (2H, m), 7.40 (1H, t, J = 15.4Hz), 7.51 (1H, t,J=18Hz),7.66(1H,m),7.83(1H,m),7.91(1H,m),8.30(1H,d,J=7.5Hz),8.92(1H,m),9.06(1H,bs),9.98(1H,s).

[0240] [Example 1]

[0241] 2-Fluoro-N-(3-Fluoro-4-(methoxymethyl)benzyl)-5-(3-(hydroxymethyl)pyridin-2-yl)benzamide

[0242] [Chemical Formula 22]

[0243]

[0244] Sodium borohydride (0.38 g, 10 mmol) was slowly added to a mixture of 2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(3-formylpyridin-2-yl)benzamide (1.0 g, 2.5 mmol) obtained in Preparation Examples 1-8 and methanol (10 mL) at 0 °C, and the mixture was stirred at room temperature for 16 hours. Water was added to the reaction mixture, and the mixture was extracted with dichloromethane (25 mL, followed by 15 mL). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure to give the title compound (0.90 g).

[0245] ESI-MS: m / z 399.50 [M+1] +

[0246] [Example 2]

[0247] Racemic 2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(3-(1-hydroxyethyl)pyridin-2-yl)benzamide

[0248] [Chemical Formula 23]

[0249]

[0250] At 0°C, methyl magnesium bromide (3M diethyl ether solution, 2.7 mL, 9.2 mmol) was added dropwise to a mixture of 2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(3-formylpyridin-2-yl)benzamide (1.3 g, 3.7 mmol) and THF (15 mL), as described in Examples 1-8, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was cooled to 0°C, and a saturated aqueous solution of ammonium chloride (5 mL) was added at the same temperature. The reaction mixture was extracted with ethyl acetate (25 mL, followed by 15 mL). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (70% ethyl acetate-n-hexane solution) to give the title compound (0.23 g).

[0251] 1H-NMR spectrum (DMSO-d6) δ (ppm): 1.23 (3H, d, J = 6.4Hz), 3.28 (3H, s), 4.42 (2H, s), 4.48 (2H, d, J = 6Hz), 4.77-4.81 (1H, m), 5.30 (1H, d, J = 4Hz), 7.1 3-7.18(2H,m),7.36-7.46(3H,m),7.64-7.67(1H,m),7.72-7.75(1H,m ),8.03-8.04(1H,d,J=6.4Hz),8.52-8.54(1H,m),9.03(1H,t,J=6Hz).

[0252] [Examples 3 and 4]

[0253] Chiral 2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(3-(1-hydroxyethyl)pyridin-2-yl)benzamide

[0254] [Chemical Formula 24]

[0255]

[0256] For the racemic 2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(3-(1-hydroxyethyl)pyridin-2-yl)benzamide (0.80 g) obtained in Example 2, the mirror-image isomers were separated using supercritical fluid chromatography (SFC) (Chiralpak-IG, 20% methanol-carbon dioxide containing 0.2% triethylamine). An early-dissolving mirror-image isomer (0.26 g, Example 3) and a late-dissolving mirror-image isomer (0.32 g, Example 4) were obtained.

[0257] [Manufacturing Example 5-1]

[0258] 4-((difluoromethoxy)methyl)-3-fluorobenzonitrile

[0259] [Chemical Formula 25]

[0260]

[0261] At room temperature, a mixture of 4-(hydroxymethyl)-3-fluorobenzonitrile (1.0 g, 6.6 mmol) and dichloromethane (5 mL) was successively mixed with 2N sodium hydroxide aqueous solution (0.53 g, 13 mmol) and (bromodifluoromethyl)trimethylsilane (1.6 mL, 9.9 mmol), and stirred at the same temperature for 48 hours. Water was added to the reaction mixture, and the mixture was extracted twice with dichloromethane (50 mL). The organic layers were combined, washed with saturated brine, and dried over sodium sulfate. The solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (2% ethyl acetate-n-hexane solution) to give the title compound (0.25 g).

[0262] 1 1H-NMR spectrum (DMSO-d6) δ (ppm): 5.05 (2H, s), 6.82 (1H, t, J = 75 Hz), 7.69-7.76 (2H, m), 7.91 (1H, d, J = 9.2 Hz).

[0263] [Manufacturing Example 5-2]

[0264] (4-((difluoromethoxy)methyl)-3-fluorophenyl)methylamine

[0265] [Chemical Formula 26]

[0266]

[0267] At 0°C, nickel(II) chloride hexahydrate (39 mg, 0.30 mmol) and sodium borohydride (0.17 g, 4.5 mmol) were slowly added to a mixture of 4-((difluoromethoxy)methyl)-3-fluorobenzonitrile (0.30 g, 1.5 mmol) and methanol (5 mL) obtained in Preparation Example 5-1, and the mixture was stirred at the same temperature for 1 hour. Ice water was added to the reaction mixture, and the mixture was extracted with dichloromethane (25 mL, twice). The two organic layers were combined, washed with saturated brine, and dried over sodium sulfate. The solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (8% methanol-dichloromethane solution) to give the title compound (200 mg).

[0268] 1 1H-NMR spectrum (DMSO-d6) δ (ppm): 3.72 (2H, s), 4.91 (2H, s), 6.77 (1H, t, J = 75 Hz), 7.15-7.23 (2H, m), 7.37-7.41 (1H, m).

[0269] [Manufacturing Example 5-3]

[0270] 5-(3-acetylpyridin-2-yl)-2-fluorobenzoic acid

[0271] [Chemical Formula 27]

[0272]

[0273] Potassium carbonate (4.5 g, 33 mmol) was added at room temperature to a mixture of 3-carboxy-4-fluorophenylboronic acid (3.0 g, 16 mmol), 3-acetyl-2-bromopyridine (3.3 g, 16 mmol), 1,4-dioxane (30 mL), and water (10 mL), followed by degassing with nitrogen. At the same temperature, a (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride dichloromethane complex (0.67 g, 0.82 mmol) was added to the reaction mixture, and the mixture was stirred at 100 °C for 16 hours. The reaction mixture was brought to room temperature, and the solvent was removed by distillation under reduced pressure. Dichloromethane (75 mL) was added to the residue, the mixture was filtered, and the residue was washed with dichloromethane (75 mL). The solvent in the filtrate was removed by distillation under reduced pressure, and the residue was purified by silica gel column chromatography (10%–12% methanol-dichloromethane solution) to give the title compound (2.1 g).

[0274] 1 1H-NMR spectrum (DMSO-d6) δ (ppm): 2.37 (3H, s), 7.28-7.31 (1H, m), 7.37-7.42 (1H, m), 7.70-7.72 (1H, m), 7.98-8.00 (1H, m), 8.10-8.12 (1H, m), 8.76-8.77 (1H, m), 13.34 (1H, bs).

[0275] [Manufacturing Example 5-4]

[0276] 5-(3-acetylpyridin-2-yl)-N-(4-((difluoromethoxy)methyl)-3-fluorobenzyl)-2-fluorobenzamide

[0277] [Chemical Formula 28]

[0278]

[0279] Under a nitrogen atmosphere and at 0°C, diisopropylethylamine (0.21 mL, 1.2 mmol) and propane phosphoric anhydride (50% ethyl acetate solution) (0.24 mL, 0.78 mmol) were added sequentially to a mixture of 5-(3-acetylpyridin-2-yl)-2-fluorobenzoic acid (0.10 g, 0.39 mmol) and DMF (3 mL) obtained in Preparation Example 5-3, and the mixture was stirred at the same temperature for 15 minutes. At the same temperature, (4-((difluoromethoxy)methyl)-3-fluorophenyl)methylamine (79 mg, 0.39 mmol) obtained in Preparation Example 5-2 was added to the reaction mixture, and the mixture was stirred at room temperature for 16 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL, twice). The two organic layers were combined, washed with saturated brine, and then dried with sodium sulfate. The solvent was removed by distillation under reduced pressure, and the residue was purified by silica gel column chromatography (55% ethyl acetate-n-hexane solution) to obtain the title compound (60 mg).

[0280] 1 H-NMR spectrum (DMSO-d6) δ (ppm): 2.38 (3H, s), 4.02 (2H, d, J = 5.6Hz), 4.93 (2H, s), 6.71 (1H, t, J = 74Hz), 7.18-7.2 1(1H,m),7.44-7.79(4H,m),7.78-7.79(1H,m),8.11(1H,d,J=7.6Hz),8.76-8.77(1H,m),9.05-9.06(1H,m).

[0281] [Example 5]

[0282] N-(4-((difluoromethoxy)methyl)-3-fluorobenzyl)-2-fluoro-5-(3-(1-hydroxyethyl)pyridin-2-yl)benzamide

[0283] [Chemical Formula 29]

[0284]

[0285] Sodium borohydride (26 mg, 0.69 mmol) was slowly added to a mixture of 5-(3-acetylpyridin-2-yl)-N-(4-((difluoromethoxy)methyl)-3-fluorobenzyl)-2-fluorobenzamide (0.15 g, 0.34 mmol) obtained in Preparation Example 5-4 and methanol (3 mL) at 0 °C, and the mixture was stirred at room temperature for 6 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL, twice). The two organic layers were combined, washed with saturated brine, and dried over sodium sulfate. The solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (55% ethyl acetate-n-hexane solution) to give the title compound (30 mg).

[0286] 1 H-NMR spectrum (DMSO-d6) δ (ppm): 1.27 (3H, d, J = 6.4Hz), 4.49 (2H, d, J = 5.6Hz), 4.77-4.81 (1H, m), 4.93 (2H, s), 5.30 (1H, J = 4Hz), 6.78 (1H, t, J = 75Hz) ,7.18-7.21(2H,m),7.39-7.47(3H,m),7.64-7.68(1H,m),7.73-7.75(1 H, m), 8.03 (1H, d, J = 8Hz), 8.53 (1H, d, J = 4.8Hz), 9.05 (1H, t, J = 5.6Hz).

[0287] [Manufacturing Example 6-1]

[0288] 2-Bromo-3-(methoxymethyl)pyridine

[0289] [Chemical Formula 30]

[0290]

[0291] A mixture of (2-bromopyridin-3-yl)methanol (0.30 g, 1.6 mmol) and THF (3 mL) was added to a mixture of 60% sodium hydride (0.057 g, 2.4 mmol) and THF (3 mL) at 0 °C. Iodomethane (0.15 mL, 2.4 mmol) was added to the reaction mixture at room temperature, and the mixture was stirred at the same temperature for 16 hours. The reaction mixture was cooled to 0 °C, ice-cold water was added, and the mixture was extracted with ethyl acetate (15 mL, followed by 5 mL). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure to give the title compound (0.15 g) as crude. This compound was used in subsequent reactions without further purification.

[0292] [Example 6]

[0293] 2-Fluoro-N-(3-Fluoro-4-(methoxymethyl)benzyl)-5-(3-(methoxymethyl)pyridin-2-yl)benzamide

[0294] [Chemical Formula 31]

[0295]

[0296] A mixture of 2-bromo-3-(methoxymethyl)pyridine (49 mg) obtained in Preparation Example 6-1, 2-fluoro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)benzamide (0.10 g), sodium carbonate (0.050 g, 0.47 mmol), 1,4-dioxacyclohexane (3 mL), and water (0.5 mL) was degassed using argon. At the same temperature, a (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloromethane dichloride complex (8.0 mg, 0.0098 mmol) was added to the reaction mixture, followed by another argon degassed reaction. The reaction mixture was stirred at 80 °C for 16 hours. The reaction mixture was brought to room temperature, water (5 mL) was added, and the mixture was extracted with ethyl acetate (10 mL, then 5 mL). The two organic layers were combined, dried with sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (20% ethyl acetate-n-hexane solution) to give the title compound (0.080 g).

[0297] 1 1H-NMR spectrum (DMSO-d6) δ (ppm): 3.28 (3H,s), 3.29 (3H,s), 4.36 (2H,s), 4.42 (2H,s), 4.48 (2H,d, J=6Hz), 7.13-7.18 (2H,m), 7.36-7.45 (3H,m), 7.75-7.79 (1H,m), 7.87-7.89 (1H,m), 7.91-7.93 (1H,m), 8.61-8.63 (1H,m), 9.01 (1H,t, J=11Hz).

[0298] [Manufacturing Example 7-1]

[0299] (5-Iodo-1H-pyrazol-4-yl)methanol

[0300] [Chemical Formula 32]

[0301]

[0302] A borane-THF solution (1M THF solution, 30 mL) was added dropwise to a mixture of ethyl 5-iodo-1H-pyrazole-4-carboxylate (4.0 g, 15 mmol) and THF (4 mL) at 0 °C. The reaction mixture was stirred at room temperature for 1 hour. Methanol (20 mL) was added dropwise to the reaction mixture at the same temperature, and the mixture was stirred for 30 minutes. The solvent was removed from the reaction mixture by distillation under reduced pressure. The residue was purified by high performance liquid chromatography to give the title compound (1.5 g).

[0303] ESI-MS: m / z 224.9 [M+1] +

[0304] [Manufacturing Example 7-2]

[0305] (3-Fluoro-4-(methoxymethyl)phenyl)methylamine

[0306] [Chemical Formula 33]

[0307]

[0308] To a mixture of 3-fluoro-4-(methoxymethyl)benzonitrile (8.0 g, 48 mmol) and methanol (50 mL) as described in Examples 1-4, Raney nickel (20 g) was added, and the mixture was stirred overnight at room temperature under a hydrogen atmosphere. The reaction mixture was degassed with nitrogen and filtered using diatomaceous earth. The solvent was removed from the filtrate by distillation under reduced pressure, yielding the title compound (7.5 g) as crude. This crude compound was used directly in subsequent reactions.

[0309] [Manufacturing Example 7-3]

[0310] 5-Bromo-2-chloro-N-(3-fluoro-4-(methoxymethyl)benzyl)benzamide

[0311] [Chemical Formula 34]

[0312]

[0313] A mixture of 5-bromo-2-chlorobenzoic acid (1.9 g, 8.1 mmol) and thionyl chloride (15 mL) was heated under reflux for 1.5 hours. The reaction mixture was brought to room temperature, and the solvent was removed by distillation under reduced pressure. At 0 °C, a mixture of crude (3-fluoro-4-(methoxymethyl)phenyl)methylamine (1.4 g), triethylamine (1.2 g, 12 mmol), and dichloromethane (15 mL) obtained in Preparation Example 7-2 was added dropwise to a mixture of the residue and dichloromethane (30 mL). The reaction mixture was stirred overnight at room temperature. Water was added to the reaction mixture at room temperature, and the mixture was extracted with dichloromethane (50 mL, 3 times). The organic layer was washed with saturated brine, dried over sodium sulfate, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound (1.1 g).

[0314] ESI-MS: m / z 386.0 [M+1] +

[0315] [Manufacturing Example 7-4]

[0316] 2-Chloro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)benzamide

[0317] [Chemical Formula 35]

[0318]

[0319] At room temperature, potassium acetate (0.84 g, 8.5 mmol) and (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (0.21 g, 0.28 mmol) were added to a mixture of 5-bromo-2-chloro-N-(3-fluoro-4-(methoxymethyl)benzyl)benzamide (1.1 g, 2.8 mmol), bis(pinacolyl)diborone (0.87 g, 3.4 mmol), and 1,4-dioxane (20 mL). The reaction mixture was stirred at 95 °C for 3 hours. The reaction mixture was brought to room temperature, water was added, and the mixture was extracted with ethyl acetate (50 mL, 3 times). The organic layer was washed with saturated brine and dried over sodium sulfate. The solvent was removed by distillation under reduced pressure to give the title compound (0.80 g).

[0320] ESI-MS: m / z 434.1 [M+1] +

[0321] [Example 7]

[0322] 2-Chloro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(4-(hydroxymethyl)-1H-pyrazol-3-yl)benzamide

[0323] [Chemical Formula 36]

[0324]

[0325] At room temperature, sodium carbonate (750 mg, 2.3 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (70 mg, 0.090 mmol) were added to a mixture of 2-chloro-N-(3-fluoro-4-(methoxymethyl)benzyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)benzamide (0.40 g, 0.92 mmol) obtained in Preparation Example 7-4, (5-iodo-1H-pyrazol-4-yl)methanol (310 mg, 1.4 mmol), n-propanol (9 mL), and water (6 mL). The mixture was stirred overnight at 95 °C. The reaction mixture was brought to room temperature, and the solvent was removed by distillation under reduced pressure. The residue was purified by high performance liquid chromatography to give the title compound (57 mg).

[0326] 1 H-NMR spectrum (CD3OD) δ (ppm): 3.40 (3H, s), 4.53 (2H, s), 4.61 (4H, d, J = 9.0 Hz), 7.20 (1H, d, J =10.9Hz),7.26(1H,d,J=7.9Hz),7.43(1H,t,J=7.6Hz),7.58(1H,s),7.70-7.99(3H,m).

[0327] [Comparative Example 1]

[0328] The following compound, which is a commercially available product, was used as Comparative Example 1. It should be noted that the following compound is the compound described in International Publication No. 2008 / 112164.

[0329] [Chemical Formula 37]

[0330]

[0331] [Comparative Example 2]

[0332] The following compound was synthesized according to the method described in International Publication No. 2015 / 127137 and used as Comparative Example 2.

[0333] [Chemical Formula 38]

[0334]

[0335] [Comparative Example 3]

[0336] The following compound was synthesized according to the method described in International Publication No. 2019 / 151241 and used as Comparative Example 3.

[0337] [Chemical Formula 39]

[0338]

[0339] [Comparative Example 4]

[0340] The following compound was synthesized according to the method described in International Publication No. 2019 / 151241 and used as Comparative Example 4.

[0341] [Chemical Formula 40]

[0342]

[0343] [Comparative Example 5]

[0344] The following compound was synthesized according to the method described in International Publication No. 2019 / 151241 and used as Comparative Example 5.

[0345] [Chemical Formula 41]

[0346]

[0347] [Comparative Example 6]

[0348] The following compound was synthesized according to the method described in International Publication No. 2019 / 151241 and used as Comparative Example 6.

[0349] [Chemical Formula 42]

[0350]

[0351] [Experimental Example 1: ALDH2 Activation]

[0352] The activation effect of the test compounds on the oxidation rate of acetaldehyde induced by ALDH2 was determined using the following method. A commercially available PicoProbe was used. TM The Aldehyde dehydrogenase Activity Assay Kit (manufactured by BioVision Inc.) was used. The experiment was conducted under the following conditions, referring to the product's instruction manual. Using a pipette, add the following (1) to (3) sequentially to each well of the 384-well plate.

[0353] (1) The recombinant human ALDH2 enzyme (manufactured by BioVision Inc.) was dissolved in ALDH assay buffer at a concentration of 3 μg / mL to obtain a solution of 10 μL.

[0354] (2) 10 μL of a 30 μM or 90 μM test compound solution (containing 3% DMSO) prepared using ALDH detection buffer.

[0355] (3) Prepare 10 μL of reaction mix according to the ratio of 8.6 μL ALDH detection buffer, 0.6 μL PicoProbe, 0.3 μL substrate mix, and 1.5 μL acetaldehyde.

[0356] Before adding (3) above, incubate at room temperature for 5 minutes. After adding (3), use EnSight (manufactured by PerkinElmer Inc.) to measure fluorescence intensity every 2.5 minutes over 180 minutes at room temperature. Fluorescence intensity was measured using an excitation wavelength of 535 nm and an emission wavelength of 587 nm.

[0357] The oxidation rates of acetaldehyde without the test compound (set to 100%) and with the test compound are shown in Table 1. The results in Table 1 confirm that the compounds in Examples 1-7 have ALDH2 activating activity.

[0358] [Table 1]

[0359] Table 1

[0360]

[0361] [Experiment 2: Metabolic stability of mouse liver microsomes was utilized]

[0362] The following methods were used to evaluate the metabolic stability of mouse liver microsomes.

[0363] 1. Materials

[0364] (1) Mouse liver microsomes: 20 mg / mL

[0365] (2) Test compound: 1.1 mM DMSO solution

[0366] (3) Potassium phosphate buffer: 66.7 mM (pH 7.4)

[0367] (4) NADPH solution: 10mM potassium phosphate buffer

[0368] (5) Quenching solution: 0.5% formic acid-acetonitrile solution with warfarin as an internal standard.

[0369] 2. Methods

[0370] 971.5 μL of potassium phosphate buffer and 27.5 μL of mouse liver microsomes were added to a propylene tube and allowed to suspend. 1 μL of the test compound was added, and 180 μL of the mixture was transferred to another tube. The mixture was pre-incubated at 37°C for 5 minutes, followed by the addition of 20 μL of NADPH solution (for a 30-minute incubation) or 20 μL of potassium phosphate buffer (for a 0-minute incubation). After incubation, 200 μL of quenching solution was added to stop the reaction. The mixture was then centrifuged at 3220 × g for 20 minutes, and the concentration of the unaltered form of the test compound in the 200 μL supernatant was determined by LC-MS / MS. Based on the peak area of ​​the unaltered form, the 0-minute incubation time was set as 100%, and the percentage of unaltered form was calculated.

[0371] 3. Results

[0372] The percentage of unchanged cells after 30 minutes is shown in Table 2. It was confirmed that the compounds of Examples 1–7 exhibited superior metabolic stability in mouse liver microsomes compared to conventional ALDH2 activators.

[0373] [Table 2]

[0374] Table 2

[0375] Survival rate (%) Example 1 90 Example 2 68 Example 3 64 Example 4 60 Example 5 52 Example 6 24 Example 7 74 Comparative Example 1 ＜1 Comparative Example 2 6 Comparative Example 3 1 Comparative Example 4 ＜1 Comparative Example 5 ＜1 Comparative Example 6 1

[0376] [Experimental Example 3: Metabolic stability of human liver microsomes was utilized]

[0377] The metabolic stability of the compounds in Examples 1, 2, 5, and 6, which had been confirmed to be stable in mouse liver microsome assays, was determined using human liver microsomes. The specific methods are described below.

[0378] 2. Methods

[0379] (1) Take 50 μL of each compound and dilute it with β-NADPH solution to obtain 0.2 μmol / L test compound.

[0380] (2) Add 50 μL of 0.2 mg protein / mL human liver microsomal solution to each sample (except for samples with a reaction time of 0 minutes).

[0381] (3) Incubate at 37°C with shaking for 30 minutes. (Concentration in the reaction sample: 0.1 μmol / L test compound, 0.1 mg protein / mL human liver microsomes)

[0382] (4) After incubation, add 400 μL of methanol to stop the reaction.

[0383] (5) Add 50 μL of 0.2 mg protein / mL liver microsome solution to the sample with a reaction time of 0 minutes.

[0384] (6) After the samples from (4) and (5) above are left to stand at -20°C for more than 30 minutes, they are centrifuged at 4°C and 3,000 rpm for about 10 minutes.

[0385] (7) The supernatant was analyzed by LC / MS / MS. Based on the peak area of ​​the unchanged body, the incubation time of 0 minutes was set as 100%, and the residual rate of the unchanged body (%) was calculated.

[0386] The percentage of unchanged bodies after 30 minutes is shown in Table 3.

[0387] [Table 3]

[0388] Table 3

[0389] Survival rate (%) Example 1 90 Example 2 87 Example 5 72 Example 6 75

[0390] [Experimental Example 4: Reactive Metabolites]

[0391] The production of glutathione conjugates was determined using LC-MS / MS in the presence of human liver microsomes, thereby investigating the risk of reactive metabolite formation from the test compounds. The specific methods are described below.

[0392] 1. Method

[0393] (1) Add 55uL of 20mg / mL human liver microsomes, 395uL of 66.7mM potassium phosphate buffer (pH 7.4), and 550uL of 10mM glutathione aqueous solution to the polypropylene tube.

[0394] (2) Add 0.55 μL of 20 mM test compound or positive control (clozapine, reloxifen, diclofenac).

[0395] (3) Divide each of the 180uL amounts into 4 tubes, label 2 as T0 and 2 as T60, and pre-incubate at 37±1℃ for 5 minutes.

[0396] (4) Add 20 μL of 10 mM NADPH solution to tube T60 and 20 μL of potassium phosphate buffer to tube T0.

[0397] (5) After 60 minutes, add 200 μL of 10% trichloroacetic acid acetonitrile solution to stop the reaction.

[0398] (6) Centrifugation was performed using a centrifuge (5810-R, Eppendorf) at 3220×g for 20 minutes. The GSH conjugate was determined by LC-MS / MS (LC: SIL-HTc, Shimazu; Mass: API-4000Qtrap, MDSsciex) for 200 μL of the supernatant.

[0399] (7) Calculate the ratio (%) of the area of ​​the GSH conjugates identified in the test compound to the area of ​​the GSH conjugates identified in clozapine.

[0400] 2. Results

[0401] The results are shown in Table 4. These results confirm that the compounds in the examples produced less reactive metabolites compared to clozapine.

[0402] [Table 4]

[0403] Table 4

[0404]

[0405] [Experimental Example 5: hERG Channel Inhibition]

[0406] The inhibitory effect of the test compound on the hERG (human Ether-a-go-go Related Gene) channel was investigated using the following patch-clamp method.

[0407] 1. Method

[0408] hERG currents passing through the entire cell membrane under constant potential were recorded using the whole-cell patch-clamp method.

[0409] To confirm the hERG current, a depolarization pulse of -80mV, -50mV, 110ms, and 20mV, 4ms was applied, followed by a repolarization pulse of -50mV, 2ms, at a frequency of once every 15 seconds. Once the obtained hERG tail current was confirmed to be stable, application began. Pulses were continued during application.

[0410] The sampling frequency is set to 5kHz, and the low-pass filter is set to 2kHz.

[0411] The temperature of the extracellular fluid in the chamber during the measurement was set at 22–25°C.

[0412] Composition of extracellular fluid

[0413] NaCl: 137 mmol / L, KCl: 4 mmol / L, CaCl2: 1.8 mmol / L, MgCl2: 1 mmol / L, D(+)-glucose: 10 mmol / L, HEPES: 10 mmol / L (adjust pH to 7.4 with 1 mol / L NaOH).

[0414] Composition of the electrolyte inside the electrode

[0415] KCl: 130 mmol / L, MgCl2: 1 mmol / L, EGTA: 5 mmol / L, Mg ATP: 5 mmol / L, HEPES: 10 mmol / L (adjust pH to 7.2 with 1 mol / L KOH).

[0416] <Fabrication of Glass Electrodes>

[0417] Glass capillary tubes (G-1.5, Seisho Scientific Instruments Co., Ltd.) were processed using a drawing apparatus (P-97, Sutter Instrument Company) to obtain tubes with a resistance value in the range of 2 to 5 MΩ when filled with electrode liquid for measurement.

[0418] <Measurement and Analysis>

[0419] For hERG current, the measurement was performed using a patch-clamp amplifier (Axopatch 200B, Molecular Devices, LLC), and the obtained electrical signal was recorded to a computer using patch-clamp recording and analysis software (pCLAMP 10, Molecular Devices, LLC).

[0420] The maximum tail current value was determined based on the current value at a depolarization pulse of -50mV and 110ms, and the rate of change (inhibition rate) after 5 minutes of application was calculated relative to the maximum tail current value before application of the test compound.

[0421] 2. Results

[0422] The results are shown in Table 5. These results confirm that the compounds in the examples do not exhibit hERG channel inhibition.

[0423] [Table 5]

[0424] Table 5

[0425]

[0426] [Experiment 6: The inhibitory effect of carrageenan-induced pain]

[0427] This experiment was conducted in accordance with Science Translational Medicine 6,2 51ra118 (2014). Male C57BL / 6J mice (Charles River Laboratories Japan, Inc.), 7 weeks old, were used. To induce allodynia in the plantar surface of the foot, carrageenan was subcutaneously applied to the left hind limb of the mice. The escape response to the mechanical stimulus was measured 180 minutes after induction using a von Frey filament with an ascending flexion force (0.16 g) applied to the plantar surface of the left hind limb. The mice were vertically stimulated for 6 seconds until the filament bent, and the escape response was rated on a 3-point scale (0: no response or startle response (moving but not lifting the foot), 1: lifting the foot, 2: licking or shaking the foot). Ten stimulations were performed, and the total score of the ten responses was calculated (escape score).

[0428] The compound (Example 1) was dissolved in DMSO / PEG400 (1:1 volume ratio). Compounds of 2, 6, or 20 mg / kg (excluding the compound in the background medium (Vehicle) group) were administered subcutaneously to the back of the neck in three equal doses at a dosage of 5 mL / kg: 15 minutes before carrageenan injection, 30 minutes after carrageenan injection, and 150 minutes after carrageenan injection. A 1.5% carrageenan solution was prepared using physiological saline and administered subcutaneously to the sole of the left hind limb at a dose of 7 μL / body.

[0429] Regarding the significance test, a Steel multiple comparison test was performed to compare the background medium group with the Example 1 group, and the significance level was calculated to be 5%. The commercially available statistical program SAS SYSTEM (SAS Software Release 9.1.3; SAS Institute Japan Ltd) was used in the significance test.

[0430] As shown in Table 6, the compounds of the examples inhibit carrageenan-induced pain.

[0431] [Table 6]

[0432] Table 6

[0433]

[0434] Each value represents the average ± SE.

[0435] Pre: Before applying carrageenan

[0436] *: p < 0.05

Claims

1. The compound represented by the following formula (1) or its pharmaceutically permissible salt, In equation (1), A is a 5- or 6-membered aromatic heterocycle containing at least one nitrogen atom as a ring member atom. R 1 and R 2 Each of the following groups is independently hydrogen, an alkyl group with 1 to 6 unsubstituted or halogenated carbon atoms, an alkenyl group with 2 to 6 unsubstituted or halogenated carbon atoms, or an alkynyl group with 2 to 6 unsubstituted or halogenated carbon atoms. R 3 It is an alkyl group with 1 to 6 unsubstituted or halogenated carbon atoms, an alkenyl group with 2 to 6 unsubstituted or halogenated carbon atoms, or an alkynyl group with 2 to 6 unsubstituted or halogenated carbon atoms. X 1 and X 2 Each is an independent halogen.

2. The compound of claim 1 or a pharmaceutically permissible salt thereof, wherein, The compound represented by formula (1) is the compound represented by formula (2) or (3) below. In the formula, R 1 R 2 R 3 X 1 and X 2 As stated above.

3. The compound of claim 1 or 2, or a pharmaceutically permissible salt thereof, wherein, R 1 It is a hydrogen-containing alkyl group, or an unsubstituted or halogen-substituted alkyl group having 1 to 6 carbon atoms.

4. The compound of claim 1 or 2, or a pharmaceutically permissible salt thereof, wherein, R 2 It is a hydrogen-containing alkyl group, or an unsubstituted or halogen-substituted alkyl group having 1 to 6 carbon atoms.

5. The compound of claim 1 or 2, or a pharmaceutically permissible salt thereof, wherein, R 3 It is an alkyl group with 1 to 6 carbon atoms, either unsubstituted or halogenated.

6. The compound of claim 1 or 2, or a pharmaceutically permissible salt thereof, wherein, X 1 It is either fluorine or chlorine.

7. The compound of claim 1 or 2, or a pharmaceutically permissible salt thereof, wherein, X 2 It is fluorine.

8. The compound of claim 1 or 2, or a pharmaceutically permissible salt thereof, wherein, The compound represented by formula (1) is selected from the group consisting of the following compounds. 。 9. A prodrug of the compound as claimed in any one of claims 1 to 8 or a pharmaceutically permissible salt thereof, wherein, R 1 It is -CH2-O-PO3H2.

10. A prodrug of the compound or a pharmaceutically permissible salt thereof as described in any one of claims 1 to 8, wherein, A contains at least one nitrogen atom as a ring member atom, wherein at least one of the nitrogen atoms is replaced by -CH2-O-PO3H2.

11. The prodrug as described in claim 9 or 10, wherein the prodrug is selected from the group consisting of compounds listed below. 。 12. An acetaldehyde dehydrogenase 2 activator comprising the compound of any one of claims 1 to 8 or a pharmaceutically permissible salt thereof, or a prodrug of any one of claims 9 to 11.

13. A pharmaceutical composition comprising any one of the compounds of claims 1 to 8 or a pharmaceutically permissible salt thereof, or any one of claims 9 to 11.

14. A treatment and / or preventive medicine for a disease selected from the group consisting of Fanconi anemia, osteoporosis, non-alcoholic fatty liver disease (NAFLD), alcoholic liver injury, pancreatitis, ischemia-reperfusion injury, peripheral artery disease, Alzheimer's disease, Parkinson's disease, esophageal cancer, head and neck cancer, and pain, wherein the treatment and / or preventive medicine comprises a compound of any one of claims 1 to 8 or a pharmaceutically permissible salt thereof, or a prodrug of any one of claims 9 to 11.

15. The treatment and / or prevention medicine for the disease as described in claim 14, wherein, The aforementioned non-alcoholic fatty liver disease, or NAFLD, is non-alcoholic steatohepatitis, or NASH.

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