Aromatic six-membered ring-fused imidazole derivatives, their preparation methods and applications

Abstract

Aromatic six-membered ring-fused imidazole derivatives, their preparation methods and applications belong to the field of pharmaceutical synthesis, and specifically relate to aromatic six-membered ring-fused imidazole derivatives represented by general formula (I), their preparation methods, and their application as LSD1 inhibitors. The compounds provided by the present invention exhibit good activity at the in vitro enzyme level. Different from the existing irreversible inhibitors with an anti-phenylcyclopropylamine parent nucleus structure, the present invention provides new skeleton compounds for the field of LSD1 inhibitor research. The raw materials used in the synthetic route adopted by the present invention are cheap and easily available, the reagents used are common reagents, the reaction conditions are mild, the post-treatment is simple, and the compounds of this type can be efficiently prepared. The compounds provided by the present invention can be used alone or in combination as lysine-specific demethylase-1 inhibitors for the treatment of various tumor diseases.

Description

Technical Field

[0001] The present invention belongs to the field of drug synthesis, and particularly relates to a novel class of aromatic six-membered ring-fused imidazole derivatives, a preparation method thereof, and their use as lysine-specific demethylase 1 inhibitors. Background Art

[0002] Epigenetics plays a key role in life, controlling gene expression and transcription, and contributing to various cellular processes, including cell differentiation, proliferation, and migration. Among various epigenetic modifications, lysine-specific demethylase 1 (LSD1) is the first identified histone demethylase, which specifically removes methyl groups in monomethylated and dimethylated H3K4 and H3K9 through enzymatic oxidation, and contributes to downstream gene transcription. By regulating gene expression, LSD1 is closely related to tumorigenesis, stem cell biology, neurodegenerative diseases, viral infections, diabetes, and fibrosis. At the same time, many research results support that the abnormal expression of LSD1 is closely related to the progression of malignant tumors such as prostate cancer, gastric cancer, breast cancer, lung cancer, and blood cancer, and inhibiting the biological function of LSD1 will produce an anti-cancer effect, indicating that LSD1 is a potential cancer treatment target. Therefore, the development of LSD1 inhibitors is beneficial to the research and development of anti-tumor drugs and can be used alone or in combination with other drugs for the treatment of cancer.

[0003] In the past few decades, great efforts have been made to develop bioactive LSD1 inhibitors, which can be divided into two types according to their mechanism of action: irreversible inhibitors and reversible inhibitors. Irreversible inhibitors can form a covalent bond with the cofactor FAD, blocking the cycle of FAD participating in demethylation, thereby exerting LSD1 inhibitory activity. So far, no LSD1 inhibitor for tumor treatment has been marketed.

[0004] Representative LSD1 inhibitors in the clinical research stage, such as ORY-1001, ORY-2001, GSK-2879552, and IMG-7289, are irreversible inhibitors with a trans-phenylcyclopropylamine as the structural core. Among the inhibitors with published structures, only Seclidemstat mesylate originally developed by the University of Utah and promoted to clinical trials by Salarius Pharmaceuticals and CC-90011 developed by Celgene Corporation are in the clinical research stage as reversible LSD1 inhibitors, and their indications are all tumors.

[0005] Currently, most of the research and development of LSD1 inhibitors are carried out with trans-phenylcyclopropylamine compounds, and developing more new skeleton LSD1 inhibitors is a research hotspot in the anti-tumor field. Summary of the Invention

[0006] The primary object of the present invention is to provide an aromatic six-membered ring-fused imidazole derivative represented by the general formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, racemate, metabolite, metabolic precursor or prodrug thereof;

[0007]

[0008] Wherein:

[0009] W and X are each independently C or N, and when one of them is C, the other can only be N;

[0010] R 1 and R 2 are each independently selected from aryl, heteroaryl, cycloalkyl, alkyl, heterocyclic group, cycloalkylalkyl, heterocyclic group alkyl, aralkyl, heteroarylalkyl; wherein, the aryl, heteroaryl, cycloalkyl, alkyl, heterocyclic group, cycloalkylalkyl, heterocyclic group alkyl, aralkyl, heteroaryl, alkyl may contain one or more substituents;

[0011] R 3 is -CH2-, carbonyl or thiocarbonyl;

[0012] R 4 is a substituted N-heterocyclic group, a substituted -N(H)-heterocyclic group alkyl, a substituted -N(Me)-heterocyclic group alkyl or -N(R 5 )2;

[0013] R 5 is selected from hydrogen, C1-C7 aliphatic primary amine, C1-C7 aliphatic secondary amine, C1-C7 aliphatic tertiary amine, preferably propylamine, N-methylpropylamine or N,N-dimethylpropylamine.

[0014] Furthermore, the aromatic six-membered ring-fused imidazole derivative represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, racemate, metabolite, metabolic precursor or prodrug thereof;

[0015] Wherein, R 5 is any one of the following structures:

[0016]

[0017] Even further, the aromatic six-membered ring-fused imidazole derivative represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, racemate, metabolite, metabolic precursor or prodrug thereof is any one of the following 46 compounds:

[0018] N-(2-Aminoethyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0019] 4-(5-(3-Methylpiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0020] 4-(5-(Morpholine-4-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0021] 4-(5-(3-Aminopiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0022] N-(3-Aminopropyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0023] 2-(4-Cyanophenyl)-N-(piperidin-4-ylmethyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0024] 2-(4-Cyanophenyl)-N-(piperidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0025] 4-(5-(4-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0026] 2-(4-Cyanophenyl)-N-(piperidin-4-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0027] 2-(4-Cyanophenyl)-N-(pyrrolidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0028] 4-(5-(4-Methylpiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0029] 4-(5-(2-Methylpiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0030] 2-(4-Cyanophenyl)-N,N-dimethyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0031] 2-(4-Cyanophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0032] 2-(4-Cyanophenyl)-N-(2-(methylamino)ethyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0033] (R)-2-(4-Cyanophenyl)-N-(pyrrolidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0034] (R)-2-(4-Cyanophenyl)-N-(piperidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0035] (R)-4-(5-(2-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0036] (S)-4-(5-(2-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0037] 4-(5-(2-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0038] N-(6-Aminohexyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0039] 2-(4-Cyanophenyl)-N-methyl-N-(2-(methylamino)ethyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0040] N-(4-Aminobutyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0041] N-(2-Aminoethyl)-2-(4-cyanophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0042] (S)-4-(5-(3-Aminopyrrolidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile

[0043] (R)-2-(4-Cyanophenyl)-N-(pyrrolidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0044] N-(2-Aminoethyl)-2-(4-fluorophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0045] N-(2-Aminoethyl)-N-methyl-2-(4-nitrophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0046] 2-([1,1'-Biphenyl]-4-yl)-N-(2-aminoethyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0047] N-(2-Aminoethyl)-N-methyl-1-(p-tolyl)-2-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazole-5-carboxamide

[0048] N-(2-Aminoethyl)-N-methyl-2-(4-(methylsulfonyl)phenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide

[0049] 4-(6-(Piperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile

[0050] 3-(4-Cyanophenyl)-N-(piperidin-4-yl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide

[0051] 4-(6-(4-Aminopiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile

[0052] N-(4-Aminobutyl)-3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide

[0053] N-(2-Aminoethyl)-3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide

[0054] 4-(6-(3-Aminopiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile

[0055] 3-(4-Cyanophenyl)-N-(piperidin-4-ylmethyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide

[0056] 3-(4-Cyanophenyl)-N-(piperidin-3-yl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide

[0057] (R)-4-(6-(3-Aminopyrrolidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile

[0058] 3-(4-Cyanophenyl)-N-(2-(methylamino)ethyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide

[0059] (R)-4-(6-(2-Methylpiperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-3-yl)benzonitrile

[0060] (S)-4-(6-(2-Methylpiperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-3-yl)benzonitrile

[0061] 4-(6-(4-Methylpiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-3-yl)benzonitrile

[0062] 4-(6-(2-Methylpiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-3-yl)benzonitrile

[0063] 3-(4-Cyanophenyl)-N-methyl-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide

[0064] In the present invention, the pharmaceutically acceptable salts of the compounds represented by the general formula (I) refer to the pharmaceutically acceptable addition salts formed by the aromatic six-membered ring-fused imidazole derivatives of the present invention and acids. The addition salts include inorganic acid and organic acid addition salts. Preferred inorganic acids and organic acids are: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid or benzoic acid.

[0065] In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine; "alkyl" refers to straight-chain or branched-chain alkyl; "alkylene" refers to straight-chain or branched-chain alkylene; "aryl" refers to an organic group obtained by removing one or two hydrogen atoms at different positions from an aromatic hydrocarbon, such as phenyl, naphthyl; "heteroaryl" refers to a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, and the ring system is aromatic, and an organic group obtained by removing one or multiple hydrogen atoms at different positions from the ring system, such as thiazolyl, imidazolyl, pyridyl, pyrazolyl, (1,2,3)- and (1,2,4)-triazolyl, furyl, thienyl, pyrrolyl, indolyl, benzothiazolyl, oxazolyl, isoxazolyl, naphthyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl; "heterocycloalkyl" refers to a monocyclic ring system containing one or more heteroatoms selected from N, O, S, such as pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, pyrazolidinyl, imidazolidinyl and oxazolinyl.

[0066] The present invention also provides a pharmaceutical composition, which contains an aromatic six-membered ring-fused imidazole derivative represented by the general formula (I), or a pharmaceutically acceptable salt thereof as an active ingredient, and is prepared by mixing with a pharmaceutically acceptable carrier or excipient, and formulated into a clinically acceptable dosage form. The pharmaceutically acceptable excipient refers to any diluent, adjuvant and / or carrier that can be used in the pharmaceutical field. The aromatic six-membered ring-fused imidazole derivative of the present invention can be used in combination with other active ingredients as long as they do not produce other toxic side effects.

[0067] The 46 aromatic six-membered ring-fused imidazole derivatives are prepared by the methods of Route 1, Route 2 and Route 3.

[0068]

[0069] Route 1: Starting material A-1 is esterified to obtain intermediate A-2. The reaction condition a is esterification under strong acid catalysis, or esterification with an acylating agent, or esterification catalyzed by dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP); the reaction solvent is methanol, ethanol or propanol, preferably methanol; the reaction temperature is 65-85 °C. Intermediate A-2 and a substituted aromatic amine or aliphatic amine undergo a nucleophilic substitution reaction to obtain intermediate A-3. The reaction condition b is carried out in the presence of an organic base or an inorganic base as an acid scavenger. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, and potassium hydroxide. Intermediate A-3 is reduced to obtain intermediate A-4. The reaction condition c is hydrogenation reduction under a palladium-based catalyst, or reduction using a combination of a reducing metal and an inorganic acid. The combination of the reducing metal and the inorganic acid includes iron powder / ammonium chloride, iron powder / hydrochloric acid, and zinc powder / acetic acid; the reaction solvent is any one or a mixture of two or more of tetrahydrofuran, methanol, ethanol, dichloromethane, and water; the preferred catalytic hydrogenation condition is 10% Pd / C as the catalyst, methanol as the solvent, and the reaction temperature is room temperature. When the reduction is carried out using a combination of a reducing metal and an inorganic acid, the iron powder / ammonium chloride system is preferred, and a mixed solvent of water and ethanol is used as the reaction solvent, and the reaction temperature is 60-80 °C. Intermediate A-4 undergoes a nucleophilic addition dehydration reaction with a substituted aromatic aldehyde or aliphatic aldehyde to obtain intermediate A-5. The reaction condition d is carried out under the catalysis of acetic acid, p-toluenesulfonic acid or a Lewis acid; the reaction solvent is tetrahydrofuran, methanol, ethanol, dichloromethane or toluene, preferably ethanol; the reaction temperature is 80-100 °C. Intermediate A-5 undergoes a cyclization reaction under iodine catalysis to obtain intermediate A-6. The reaction condition e is carried out under the catalysis of iodine, N-iodosuccinimide (NIS) or iodine chloride; the reaction solvent is methanol, ethanol, propanol, trifluoroethanol, acetone, N,N-dimethylformamide or dimethyl sulfoxide; the preferred reaction condition is N-iodosuccinimide as the catalyst, trifluoroethanol as the solvent, and the reaction is carried out at 25-60 °C. Intermediate A-6 is hydrolyzed by an ester to obtain intermediate A-7. The reaction condition f is carried out under the catalysis of an organic base or an inorganic base. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, and potassium hydroxide; the reaction solvent is a polar solvent including methanol, ethanol, propanol, acetone, N,N-dimethylformamide, and dimethyl sulfoxide; the preferred reaction condition is sodium hydroxide as the acid scavenger, ethanol as the solvent, and the reaction is carried out at 80 °C.Intermediate A-7 undergoes acylation reaction with a substituted aliphatic amine to obtain Intermediate A-8. The reaction condition g is carried out in the presence of a condensing agent such as 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-(benzotriazol-1-yl)-bis(dimethylamino)carbenium hexafluorophosphate (HBTU), O-(5-chlorobenzotriazol-1-yl)-bis(dimethylamino)carbenium hexafluorophosphate (HCTU), O-(benzotriazol-1-yl)-bis(dimethylamino)carbenium tetrafluoroborate (TBTU), O-(N-succinimidyl)-bis(dimethylamino)carbenium tetrafluoroborate (TSTU), O-(N-endo-5-norbornene-2,3-dicarboximide)-bis(dimethylamino)carbenium tetrafluoroborate (TNTU), diphenylphosphoryl chloride (DPP-Cl), diethyl cyanophosphonate (DECP), diphenylphosphoryl azide (DPPA), methylphosphonyl thioazide (MPTA), bis(2-oxo-3-oxazolidinyl)phosphoryl chloride (BOP-Cl) or 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorin 2,4,6-trioxide (T3P); the reaction solvent is a polar solvent including methanol, ethanol, propanol, acetone, N,N-dimethylformamide, dimethyl sulfoxide; the preferred reaction condition is using N,N-dimethylformamide as the solvent and HATU as the condensing agent. Intermediate A-8 is obtained by removing the Boc protecting group to obtain the target product A-9. The reaction condition h is under acid catalysis, heating for 4 - 12 h; the acid is trifluoroacetic acid, hydrochloric acid, preferably hydrochloric acid; the reaction solvent is methanol, ethanol, isopropanol, N,N-dimethylformamide, 1,4-dioxane, ethyl acetate or dimethyl sulfoxide, preferably acid / solvent is hydrochloric acid / ethyl acetate; the reaction temperature is 25 - 80 °C.

[0070] Route 2: The reaction conditions and operation methods in Route 2 adopt the corresponding reaction conditions and operation methods in Route 1.

[0071] Route 3: The starting material B-1 is esterified to obtain intermediate B-2. The reaction condition a is esterification under strong acid catalysis, or esterification with an acylating agent, or esterification catalyzed by dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP); the reaction solvent is methanol, ethanol or propanol, preferably methanol; the reaction temperature is 80 °C. Intermediate B-2 and a substituted α-halo saturated carbonyl compound undergo a cyclization reaction to obtain intermediate B-3. The reaction condition b is carried out in the presence of an organic base or an inorganic base as an acid scavenger. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide and potassium hydroxide; the reaction solvent is a polar solvent including methanol, ethanol, propanol, acetone, N,N-dimethylformamide and dimethyl sulfoxide; the preferred reaction condition is potassium carbonate as the acid scavenger, ethanol as the solvent, and reaction at 80 °C. Intermediate B-3 is iodinated to obtain intermediate B-4. The reaction condition c is iodination catalyzed by iodine, N-iodosuccinimide (NIS) or iodine monochloride; the reaction solvent is methanol, ethanol, propanol, trifluoroethanol, acetonitrile, acetone, tetrahydrofuran, N,N-dimethylformamide or dimethyl sulfoxide; the preferred reaction condition is N-iodosuccinimide as the catalyst, trifluoroethanol as the solvent, and reaction at 25-60 °C. Intermediate B-4 and a substituted arylboronic acid or aliphatic boronic acid undergo a Suzuki reaction to obtain intermediate B-5. The reaction condition d is heating under palladium complex catalysis, inorganic base and anaerobic conditions; the palladium complex is Pd(PPh3)4, PdCl2, PdCl2(dppf), Pd(OAc)2 or Pd(PPh3)2Cl2, preferably PdCl2(dppf); the inorganic base is potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate or potassium fluoride, preferably potassium carbonate; the reaction solvent is any one or a mixture of two or more of ethanol, 1,4-dioxane, tetrahydrofuran, toluene, N,N-dimethylformamide, dimethyl sulfoxide, water and ethylene glycol dimethyl ether. The preferred solvent is a mixed solvent of 1,4-dioxane and water in a volume ratio of 4:1; the reaction temperature is 80-140 °C. Intermediate B-5 undergoes an ester hydrolysis reaction to obtain intermediate B-6. The reaction condition e is carried out in the presence of an organic base or an inorganic base as a catalyst. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide and potassium hydroxide; the reaction solvent is methanol, ethanol, propanol, acetone, N,N-dimethylformamide or dimethyl sulfoxide; the preferred reaction condition is sodium hydroxide as the acid scavenger, ethanol as the solvent, and reaction at 80 °C. Intermediate B-6 and a substituted aliphatic amine undergo an acylation reaction to obtain intermediate B-7. The reaction condition f adopts the reaction condition of reaction route g in Route 1. Intermediate B-7 undergoes a reaction to remove the Boc protecting group to obtain the target product B-8. The reaction condition g adopts the reaction condition of reaction route h in Route 1.

[0072] The present invention also provides the use of the aromatic six-membered ring-fused imidazole derivatives or pharmaceutically acceptable salts thereof, or a pharmaceutical composition comprising the derivatives as a lysine-specific demethylase 1 inhibitor.

[0073] The present invention also provides the use of the aromatic six-membered ring-fused imidazole derivatives or pharmaceutically acceptable salts thereof, or a pharmaceutical composition comprising the derivatives for the preparation of a medicament for the treatment of tumor diseases.

[0074] Advantages of the present invention:

[0075] 1) As an LSD1 inhibitor, the 46 preferred aromatic six-membered ring-fused imidazole derivatives provided by the present invention exhibit good activity at the in vitro enzyme level.

[0076] 2) Different from the existing irreversible inhibitors with an anti-phenylcyclopropylamine core structure, the present invention provides new skeleton compounds for the field of LSD1 inhibitor research.

[0077] 3) The raw materials used in the synthetic route adopted by the present invention are cheap and easily available, the reagents used are common reagents, the reaction conditions are mild, the post-treatment is simple, and the compounds of this type can be efficiently prepared. Detailed implementation manners

[0078] The examples are intended to illustrate rather than limit the scope of the present invention. The 1H nuclear magnetic resonance spectrum of the compound was measured by Bruker ARX-400, and the mass spectrum was measured by Agilent 1100 LC / MSD; all the reagents used were of analytical grade or chemical pure grade.

[0079] Table 1. Structural formulas, chemical names, and relative molecular masses of the examples

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086] Example 1

[0087] Synthesis of N-(2-aminoethyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide, using the preparation route of Route 1

[0088] Step 1: Preparation of methyl 4-chloro-3-nitrobenzoate (A-2)

[0089]

[0090] 4-Chloro-3-nitrobenzoic acid (10 g, 49.6 mmol) was added to a 500 mL single-neck reaction flask, and 250 mL of methanol was added. It was stirred at room temperature until completely dissolved. Subsequently, 10 mL of concentrated sulfuric acid was slowly added dropwise to the reaction solution. After addition, the temperature was raised to 65 °C and the reaction continued for 4 h. The reaction solution was cooled to room temperature, and the reaction solution was concentrated under reduced pressure. 400 mL of water was added with stirring, and a white solid was precipitated. It was filtered by suction and the filter cake was rinsed with water and dried to obtain 10.2 g of a crude white solid with a yield of 95.4%.

[0091] Step 2: Preparation of methyl 3-nitro-4-(p-tolylamino)benzoate (A-3)

[0092]

[0093] A-2 (5.4 g, 25 mmol) and 4-methylaniline (8 g, 75 mmol) were added to a 250 mL reaction flask, and 50 mL of dimethyl sulfoxide was added. The temperature was raised to 70 °C and the reaction was carried out for 12 h. The reaction solution was cooled to room temperature, 100 mL of water was added, and an orange solid was precipitated. It was filtered by suction and the filter cake was rinsed with water and dried to obtain 6.2 g of a crude orange solid with a yield of 86.7%.

[0094] Step 3: Preparation of methyl 3-amino-4-(p-tolylamino)benzoate (A-4)

[0095]

[0096] A-3 (2 g, 7 mmol) and 10% palladium on carbon (0.2 g) were added to a 100 mL reaction flask, and 40 mL of methanol was added. Under hydrogen conditions, the reaction was carried out at room temperature for 12 h. It was filtered, and the filtrate was concentrated under reduced pressure to obtain 1.7 g of a crude pale yellow solid with a yield of 94.9%.

[0097] Step 4: Preparation of methyl (E)-3-((4-cyanobenzylidene)amino)-4-(p-tolylamino)benzoate (A-5)

[0098]

[0099] A-4 (1.5 g, 5.9 mmol) and 4-cyanobenzaldehyde (1 g, 7.6 mmol) were added to a 100 mL reaction flask, 0.5 mL of acetic acid was added dropwise, 30 mL of ethanol was added, and the temperature was raised to 80 °C and the reaction was carried out for 12 h. The reaction solution was cooled to room temperature, and the reaction solution was concentrated under reduced pressure to obtain 1.6 g of a crude yellow solid, which was directly used for the next step without treatment, with a yield of 73.5%.

[0100] Step 5: Preparation of methyl 2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxylate (A-6)

[0101]

[0102] Add A-5 (1.6 g, 4.3 mmol) to a 100 mL reaction flask, add 30 mL of trifluoroethanol, and slowly add N-iodosuccinimide (1.8 g, 8 mmol) with stirring. Heat the reaction mixture to 50 °C and react for 2 h. Concentrate the reaction solution under reduced pressure, add saturated sodium thiosulfate solution, extract with ethyl acetate (40 mL × 3), and wash with saturated sodium chloride solution (40 mL × 3). Dry the ethyl acetate phase with anhydrous sodium sulfate and concentrate under reduced pressure. Purify by silica gel column chromatography to obtain 0.95 g of a white solid with a yield of 60.2%.

[0103] Step 6: Preparation of 2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxylic acid (A-7)

[0104]

[0105] Add A-6 (5 g, 13.6 mmol) and sodium hydroxide (1.6 g, 40.8 mmol) to a 250 mL reaction flask, add 100 mL of ethanol, and react at 80 °C for 3 h. Cool the reaction solution to room temperature, concentrate the reaction solution, dilute with water, filter to remove insoluble substances, adjust the pH to 2 with 1 N hydrochloric acid, precipitate a white solid, and dry to obtain 4.3 g of a crude white solid with a yield of 89.6%.

[0106] Step 7: Preparation of tert-butyl (2-(2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamido)ethyl)carbamate (A-8)

[0107]

[0108] Add A-7 (0.35 g, 1 mmol), N-Boc-1,2-ethylenediamine (0.4 g, 2.5 mmol), and HATU (0.38 g, 1.0 mmol) to a 50 mL reaction flask, add 10 mL of anhydrous N,N-dimethylformamide and 0.19 mL of N,N-diisopropylethylamine, and react at 40 °C for 12 h. Pour the reaction solution into 50 mL of water, extract with dichloromethane (30 mL × 3), wash with saturated sodium chloride solution (30 mL × 3), combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. Purify by silica gel column chromatography to obtain 0.4 g of a white solid with a yield of 80.7%.

[0109] Step 8: Preparation of N-(2-aminoethyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide (A-9)

[0110]

[0111] Add A-8 (0.25 g, 0.5 mmol) to a 50 mL reaction flask, add 20 mL of 2N hydrochloric acid / ethyl acetate solution, and stir the reaction at room temperature for 2 h. Concentrate the reaction solution under reduced pressure, add 5 mL of water, and adjust the pH to 10 with 5N sodium hydroxide solution. Extract with dichloromethane (10 mL × 3), wash with saturated sodium chloride solution (10 mL × 3), combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 0.18 g of a white solid with a yield of 93.1%.

[0112] Examples 2-26 were prepared using the preparation method in Example 1 to prepare the corresponding compounds shown in Table 1.

[0113] Example 27

[0114] Synthesis of N-(2-aminoethyl)-2-(4-fluorophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide was carried out using the preparation route of Route 2

[0115] Step 1: Preparation of 3-nitro-4-(p-toluidine)benzoic acid (A-10)

[0116]

[0117] According to the operation method in Step 6 of Example 1, 3-nitro-4-(p-toluidine)benzoic acid (A-10) was synthesized from compound A-3.

[0118] Step 2: Preparation of tert-butyl (2-(N-methyl-3-nitro-4-(p-tolylamino)benzamido)ethyl)carbamate (A-11)

[0119]

[0120] According to the operation method in Step 7 of Example 1, tert-butyl (2-(N-methyl-3-nitro-4-(p-tolylamino)benzamido)ethyl)carbamate (A-11) was synthesized from compound A-10.

[0121] Step 3: Preparation of tert-butyl (2-(3-amino-N-methyl-4-(p-tolylamino)benzamido)ethyl)carbamate (A-12)

[0122]

[0123] According to the operation method in Step 3 of Example 1, (tert-butyl (2-(3-amino-N-methyl-4-(p-tolylamino)benzamido)ethyl)carbamate (A-12) was synthesized from compound A-11.

[0124] Step 4: Preparation of (E)-(tert-butyl (2-(3-((4-fluorobenzylidene)amino)-N-methyl-4-(p-tolylamino)benzamido)ethyl)carbamate (A-13)

[0125]

[0126] According to the operation method in Step 4 of Example 1, (E)-(tert-butyl (2-(3-((4-fluorobenzylidene)amino)-N-methyl-4-(p-tolylamino)benzamido)ethyl)carbamate (A-13) was synthesized from compound A-12.

[0127] Step 5: Preparation of (tert-butyl (2-(2-(4-fluorophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamido)ethyl)carbamate (A-14)

[0128]

[0129] According to the operation method in Step 5 of Example 1, (tert-butyl (2-(2-(4-fluorophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamido)ethyl)carbamate (A-14) was synthesized from compound A-13.

[0130] Step 6: Preparation of N-(2-aminoethyl)-2-(4-fluorophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide (A-15)

[0131]

[0132] According to the operation method in Step 8 of Example 1, N-(2-aminoethyl)-2-(4-fluorophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide (A-15) was synthesized from compound A-14.

[0133] In Examples 28 - 31, the corresponding compounds shown in Table 1 were prepared using the preparation method in Example 27.

[0134] Example 32

[0135] Synthesis of 4-(6-(piperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile was carried out using the preparation route of Route 3.

[0136] Step 1: Preparation of methyl 6-aminonicotinate (B-2)

[0137]

[0138] According to the operation method of Step 1 in Example 1, methyl 6-aminonicotinate (B-2) was synthesized from 6-aminonicotinic acid.

[0139] Step 2: Preparation of methyl 2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxylate (B-3)

[0140]

[0141] B-2 (10 g, 65.8 mmol), 2-bromo-4'-methylacetophenone (13.9 g, 65.8 mmol) and potassium carbonate (9.1 g, 65.8 mmol) were added to a 500 mL reaction flask, 200 mL of ethanol was added, and the reaction was carried out at 80 °C for 5 h. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 9.8 g of a yellow solid with a yield of 56%.

[0142] Step 3: Preparation of methyl 3-iodo-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxylate (B-4)

[0143]

[0144] B-3 (8 g, 30.1 mmol) was added to a 250 mL reaction flask, 100 mL of acetonitrile was added, and N-iodosuccinimide (7.4 g, 32.9 mmol) was slowly added with stirring at room temperature for 1 h. The reaction solution was concentrated under reduced pressure, the reaction solution was poured into 100 mL of water, a yellow solid was precipitated, filtered by suction and the filter cake was rinsed with water and dried to obtain 11.3 g of a crude yellow solid with a yield of 95.8%.

[0145] Step 4: Preparation of methyl 3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxylate (B-5)

[0146]

[0147] B-4 (5 g, 12.8 mmol), Pd(dppf)Cl2 (0.5 g, 0.7 mmol), potassium carbonate (5.3 g, 38.4 mmol) and 4-cyanophenylboronic acid (2.3 g, 15.6 mmol) were added into a 100 mL reaction flask, and 30 mL of dioxane and 7.5 mL of water were added. The reaction was carried out at 90 °C for 12 h under argon protection. The reaction solution was cooled and concentrated, 40 mL of water was added, and it was extracted with ethyl acetate (40 mL × 3) and washed with saturated brine (40 mL × 3). The ethyl acetate phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Purification by silica gel column chromatography gave 3.4 g of a yellow solid with a yield of 72.3%.

[0148] Step 5: Preparation of 3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxylic acid (B-6)

[0149]

[0150] According to the operation method of Step 6 in Example 1, 3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxylic acid (B-6) was synthesized from compound B-5.

[0151] Step 6: Preparation of tert-butyl 4-(3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carbonyl)piperazine-1-carboxylate (B-7)

[0152]

[0153] According to the operation method of Step 7 in Example 1, tert-butyl 4-(3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carbonyl)piperazine-1-carboxylate (B-7) was synthesized.

[0154] Step 7: Preparation of 4-(6-(piperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-3-yl)benzonitrile (B-8)

[0155]

[0156] According to the operation method of Step 8 in Example 1, 4-(6-(piperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-3-yl)benzonitrile (B-8) was synthesized from compound B-7.

[0157] In Examples 33 - 46, the corresponding compounds shown in Table 1 were prepared by the preparation method in Example 32.

[0158] In vitro pharmacological tests of some products of the present invention

[0159] Test 1:

[0160] The 700120 kit from Cayman Company was purchased in this invention to detect the LSD1 inhibitory activity of all compounds in the above Examples 1 - 46. The specific operation steps are as follows:

[0161] (1) Detection settings include 100% activity wells, background wells, positive control wells, and compound wells. Each group has three replicate wells.

[0162] (2) 100% activity wells: Add 120 μL of LSD1 Buffer solution, 10 μL of solution (the same composition as the solution for dissolving the compound and the positive drug), 20 μL of LSD1 enzyme, and 20 μL of LSD1 detection peptide in sequence.

[0163] (3) Test wells and positive control wells: Add 120 μL of LSD1 Buffer solution, 10 μL of the test compound solution, 20 μL of LSD1 enzyme, and 20 μL of LSD1 detection peptide in sequence.

[0164] (4) Background wells: Add 140 μL of LSD1 Buffer solution, 10 μL of solution (the same composition as the solution for dissolving the compound and the positive drug), and 20 μL of LSD1 enzyme in sequence.

[0165] (5) During the process of adding the solution, place the 96 - well plate on an ice pack to cool down and prevent the enzymatic reaction. After adding, incubate at room temperature for 30 min in the dark.

[0166] (6) After incubating for 30 min, add 20 μL of horseradish peroxidase solution and 10 μL of fluorescent substrate solution to each well in sequence. Incubate at room temperature for 10 min in the dark.

[0167] (7) Excite at a wavelength of 530 nM with an enzyme - linked immunosorbent assay (ELISA) reader and detect the fluorescence intensity of the emission at 590 nM.

[0168] Inhibition rate % = (100% activity well - sample well) / 100% activity well * 100

[0169] Table 2. 1H NMR and enzyme inhibition activity data of the compounds prepared in Examples 1 - 46

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179] In the present invention, the aromatic six-membered ring-fused imidazole derivatives represented by the general formula (I) and their pharmaceutically acceptable salts can be administered alone, but are usually administered in a mixture with a pharmaceutical carrier. The selection of the pharmaceutical carrier should be based on the desired route of administration and standard pharmaceutical practice. The following will illustrate their new applications in the pharmaceutical field by the preparation methods of various pharmaceutical dosage forms of such compounds, such as tablets, capsules, injections, aerosols, suppositories, membranes, dripping pills, external liniments, and ointments.

[0180] Test Two:

[0181] In vitro H + ,K + -ATPase Pharmacological Test

[0182] In this experiment, enzyme standard products were extracted in the manner reported in the literature for in vitro enzyme activity tests. The specific operation steps are as follows: Prepare the gastric mucosal microsomal fraction from fresh pig stomach. First, rinse the removed stomach under tap water to remove impurities, and then immerse it in 3 mol / L NaCl solution to remove mucus. Separate the gastric mucosa from the stomach surface and chop it into a buffer solution composed of 1 mmol / L EDTA, 10 mmol / L Tris-HCl, and 0.25 mol / L sucrose solution, and then homogenize for 10 min. Centrifuge the obtained homogenate at 20000 g for 30 min, and further centrifuge the supernatant at 100000 g for 90 min. Suspend the precipitate evenly in the buffer solution, overlay it on 0.25 mol / L sucrose solution containing 7.5% Ficoll, and centrifuge at 100000 g for 90 min. Collect the microsomal fraction and wash it by centrifugation with 0.25 mol / L sucrose solution. Then disperse it in the buffer solution, and determine the protein concentration in the suspension by the Brandford method, which is used as the standard product of H + ,K + -ATPase.

[0183] Determine H + ,K +-ATPase activity. The enzyme standard was diluted to a concentration of 2.5 μg / mL in 50 mmol / L HEPES Tris buffer (5 mmol / L magnesium chloride, 10 mmol / L potassium chloride, 10 μmol / L valine, pH 6.5). 5 μL of the test compound dissolved in different concentrations of 10% DMSO solution was added to the diluted enzyme solution, and then incubated at 37 °C for 30 min. Subsequently, 5 μL of 2 mmol / L adenosine triphosphate Tris salt buffer (50 mmol / L HEPES Tris buffer (5 mmol / L magnesium chloride, pH 6.5)) was added, and an enzymatic reaction was carried out at 37 °C for 20 min. The reaction was terminated by adding 15 μL of malachite green reagent (0.12% malachite green in sulfuric acid (2.5 mol / L), 7.5% ammonium molybdate, and 11% Tween 20, reagent ratio 100:25:2), and allowed to stand at room temperature for 15 min. Finally, the reaction product of inorganic phosphorus and malachite green in the mixture was measured at a wavelength of 610 nm. The activity of H + ,K + -ATPase was calculated based on the difference in activity with and without potassium chloride. The inhibition of the test compound was expressed as a percentage, and the corresponding IC 50 value was calculated using GraphPad Prism.

[0184] Table 3. In vitro H + ,K + -ATPase inhibitory activity of compounds in some examples at a concentration of 10 μM

[0185] Example Inhibition rate Example Inhibition rate 3 36.55％ 18 23.17％ 4 20.36％ 20 24.82％ 6 22.73％ 21 35.34％ 7 27.85％ 22 30.63％ 9 43.58％ 23 31.86％ 11 32.61％ 24 24.73％ 12 18.23％ 26 49.67％ 13 36.01％ 27 41.06％ 15 35.34％ 36 38.45％

[0186] Test three:

[0187] In vivo gastric acid secretion inhibition test of some products of the present invention in rats

[0188] The candidate drug was stored at 4 °C for a long time and prepared immediately before use. When dissolving, anhydrous ethanol was added first, and after complete dissolution, ultrapure water was slowly added multiple times until the final ethanol concentration in the solution was 10%. The dosing volume of each group of candidate drug, solvent, and modeling agent was 10 mL / kg.

[0189] (1) Effects of the test compound on the basal gastric acid secretion in SD rats: Male SD rats weighing 250 - 280 g were selected. After all the rats were weighed and numbered, they were placed in a fasting cage for 24 - hour fasting, and were allowed to drink water freely during the fasting period. After fasting, they were weighed again and grouped according to body weight for the experiment. Starting from the time of drug administration, the candidate drug or solvent was administered by gavage, and the time when the administration was completed was recorded. Thereafter, food and water were withheld. One hour later, the rats were anesthetized by intraperitoneal injection of 300 mg / kg chloral hydrate. After anesthesia, the rats were fixed on the operating board with their chests facing up and their body temperature was maintained with a thermostatic heating pad. The peritoneum was carefully incised with a scalpel to expose the abdominal cavity, the stomach and pylorus were found and carefully removed. After ligation at the pyloric ring, the gastric body was replaced and the abdominal cavity was sutured; the rats were placed on their sides in an incubator at 37 °C until they woke up, and after waking up, they were transferred to a fasting environment. During this period, the physiological status of the rats was observed every 30 minutes, and the excreta were cleaned up in a timely manner; 3 hours after ligation, the rats were sacrificed by overdose anesthesia with urethane, the abdominal cavity was opened again and the cardia was ligated, and the gastric body was removed by cutting the esophagus and duodenum with surgical scissors; a syringe was used to pierce the gastric body on the lesser curvature side of the stomach and 5 mL of normal saline was injected to fully mix the gastric contents with it, and then the mixture was collected. The residue was scraped off the gastric wall with a steel spatula and the steel spatula was washed with normal saline, and the washing solution was transferred into a centrifuge tube together. The final mixed liquid was centrifuged at 3000 r / min (1500 g) for 10 minutes. The supernatant was taken into a conical flask, phenolphthalein indicator was added, and titrated with 0.01000 mol / L NaOH standard solution, and the gastric acid secretion volume within 3 hours was calculated and statistically analyzed.

[0190] (2) Effect of the test compound on histamine-stimulated gastric acid secretion in SD rats: Male SD rats weighing 250 - 280 g were selected. After all the rats were weighed and numbered, they were placed in a fasting cage for 24 h of fasting, and were allowed free access to water during fasting. After fasting, they were weighed again and grouped according to body weight for the experiment. Starting from the time of administration, the candidate drug or solvent was administered by gavage, and the time when the administration was completed was recorded. Thereafter, food and water were withheld. 1 h later, the rats were anesthetized by intraperitoneal injection of 300 mg / kg chloral hydrate. After anesthesia, the rats were fixed on the operating board with their chests facing up and their body temperature was maintained with a thermostatic heating pad. The peritoneum was carefully incised with a scalpel to expose the abdominal cavity, the stomach and pylorus were found and carefully removed. After ligation at the pyloric ring, the gastric body was replaced and the abdominal cavity was sutured, and 30 mg / kg of histamine was injected subcutaneously; the rats were placed on their sides in an incubator at 37 °C until they woke up, and after waking up, they were transferred to a fasting environment. During this period, the physiological status of the rats was observed every 30 min, and the excreta were cleaned up in a timely manner; 3 h after ligation, the rats were sacrificed by overdose anesthesia with urethane, the abdominal cavity was opened again and the cardia was ligated, and the gastric body was removed by cutting the esophagus and duodenum with surgical scissors; a syringe was used to pierce the gastric body on the lesser curvature side of the stomach and 5 mL of normal saline was injected to fully mix the gastric contents with it, and the mixture was collected. The residue was scraped off the gastric wall with a steel spatula and the steel spatula was washed with normal saline, and the washing solution was transferred to a centrifuge tube together. The final mixed liquid was centrifuged at 3000 r / min (1500 g) for 10 min. The supernatant was taken into a conical flask, phenolphthalein indicator was added, and titration was carried out with 0.01000 mol / L NaOH standard solution, and the gastric acid secretion volume within 3 h was calculated and statistically analyzed.

[0191] (3) Effects of the test compound on gastric acid secretion stimulated by 2-deoxy-D-glucose in SD rats: Male SD rats weighing 250 - 280 g were selected. After all the rats were weighed and numbered, they were placed in fasting cages for 24 h of fasting, and were allowed free access to water during fasting. After fasting, they were weighed again and grouped according to body weight for the experiment. Starting from the time of drug administration, the candidate drug or solvent was administered by gavage, and the time when the administration was completed was recorded. Thereafter, food and water were withheld. 1 h later, the rats were anesthetized by intraperitoneal injection of 300 mg / kg chloral hydrate. After anesthesia, the rats were fixed on the operating board with their chests facing up and their body temperature was maintained with a thermostatic heating pad. The peritoneum was carefully incised with a scalpel to expose the abdominal cavity, the stomach and pylorus were found and carefully removed. After ligation at the pyloric ring, the gastric body was replaced and the abdominal cavity was sutured, and 2-deoxy-D-glucose at 200 mg / kg was injected subcutaneously; the rats were placed on their sides in an incubator at 37 °C until they woke up, and then transferred to a fasting environment. During this period, the physiological state of the rats was observed every 30 min, and the excreta were cleaned up in a timely manner; 3 h after ligation, the rats were sacrificed by overdose anesthesia with urethane, the abdominal cavity was opened again and the cardia was ligated, and the gastric body was removed by cutting the esophagus and duodenum with surgical scissors; a syringe was used to pierce the gastric body on the lesser curvature side of the stomach and 5 mL of normal saline was injected to mix the gastric contents thoroughly, and then the mixture was collected. The residue was scraped off along the gastric wall with a steel spatula and the spatula was washed with normal saline, and the washing solution was transferred into a centrifuge tube together. The final mixed liquid was centrifuged at 3000 r / min (1500 g) for 10 min. The supernatant was taken into a conical flask, phenolphthalein indicator was added, and titrated with 0.01000 mol / L NaOH standard solution, and the gastric acid secretion volume within 3 h was calculated and statistically analyzed.

[0192] Table 4. Inhibitory activity of compounds in some examples on gastric acid secretion in vivo at different concentrations

[0193]

[0194] Example 47

[0195] Tablet

[0196] It contains 10 g of the aromatic six-membered ring and imidazole derivative prepared in Example 1. After mixing 20 g of excipients according to the general tabletting method in pharmacy, it was pressed into 100 tablets, and each tablet weighed 300 mg.

[0197] Example 48

[0198] Capsule

[0199] It contains 10 g of the aromatic six-membered ring and imidazole derivative prepared in Example 2. After mixing 20 g of excipients according to the requirements of pharmacy for capsules, it was filled into hollow capsules, and each capsule weighed 300 mg.

[0200] Example 49

[0201] Injection

[0202] Containing 10 g of the aromatic six-membered ring-fused imidazole derivative prepared in Example 3, activated carbon adsorption was carried out according to the conventional pharmaceutical method. After filtration through a 0.65 μm microporous membrane, it was filled into a nitrogen tank to prepare an aqueous injection preparation, with each vial containing 2 mL and a total of 100 vials filled.

[0203] Example 50

[0204] Aerosol

[0205] Containing 10 g of the aromatic six-membered ring-fused imidazole derivative prepared in Example 1, after dissolving it with an appropriate amount of propylene glycol, adding distilled water and other excipients, a 500 mL clear solution was prepared.

[0206] Example 51

[0207] Suppository

[0208] Containing 10 g of the aromatic six-membered ring-fused imidazole derivative prepared in Example 1, it was ground fine and an appropriate amount of glycerol was added, and after grinding evenly, the melted glycerogelatin was added and ground evenly. Then it was poured into a lubricated mold to prepare 50 suppositories.

[0209] Example 52

[0210] Film

[0211] Polyvinyl alcohol, medicinal glycerol, and water were stirred and swollen and then heated to dissolve. After filtration through an 80-mesh sieve, 10 g of the aromatic six-membered ring-fused imidazole derivative prepared in Example 18 was added to the filtrate and stirred to dissolve. 100 films were prepared by a film coating machine.

[0212] Example 53

[0213] Drop pill

[0214] Containing 10 g of the aromatic six-membered ring-fused imidazole derivative prepared in Example 5, after heating and melting and mixing evenly with 50 g of matrix such as gelatin, it was dropped into low-temperature liquid paraffin to prepare a total of 1000 drop pills.

[0215] Example 54

[0216] Topical liniment

[0217] Containing 10 g of the aromatic six-membered ring-fused imidazole derivative prepared in Example 5, it was mixed and ground with 2.5 g of excipients such as emulsifier according to the conventional pharmaceutical method, and then distilled water was added to 200 mL to prepare it.

[0218] Example 55

[0219] Ointment

[0220] It is prepared by grinding 10 g of the aromatic six-membered ring-fused imidazole derivative obtained in Example 35 and uniformly grinding it with 500 g of an oily matrix such as petrolatum.

[0221] Although the present invention has been described by specific embodiments, modifications and equivalent variations will be apparent to those skilled in the art, and they are all included in the scope of the present invention.

Claims

1. An aromatic six-membered ring-fused imidazole derivative or a pharmaceutically acceptable salt thereof, characterized in that, The aromatic six-membered ring-fused imidazole derivatives shown are any one of the following 46 compounds: N-(2-Aminoethyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 4-(5-(3-Methylpiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile 4-(5-(Morpholine-4-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile 4-(5-(3-Aminopiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile N-(3-Aminopropyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 2-(4-Cyanophenyl)-N-(piperidin-4-ylmethyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 2-(4-Cyanophenyl)-N-(piperidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 4-(5-(4-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile 2-(4-Cyanophenyl)-N-(piperidin-4-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 2-(4-Cyanophenyl)-N-(pyrrolidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 4-(5-(4-Methylpiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile 4-(5-(2-Methylpiperidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile 2-(4-Cyanophenyl)-N,N-dimethyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 2-(4-Cyanophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 2-(4-Cyanophenyl)-N-(2-(methylamino)ethyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide (R)-2-(4-Cyanophenyl)-N-(pyrrolidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide (R)-2-(4-Cyanophenyl)-N-(piperidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide (R)-4-(5-(2-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile (S)-4-(5-(2-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile 4-(5-(2-Methylpiperazine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile N-(6-Aminohexyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 2-(4-Cyanophenyl)-N-methyl-N-(2-(methylamino)ethyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide N-(4-Aminobutyl)-2-(4-cyanophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide N-(2-Aminoethyl)-2-(4-cyanophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide (S)-4-(5-(3-Aminopyrrolidine-1-carbonyl)-1-(p-tolyl)-1H-benzo[d]imidazol-2-yl)benzonitrile (R)-2-(4-Cyanophenyl)-N-(pyrrolidin-3-yl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide N-(2-Aminoethyl)-2-(4-fluorophenyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide N-(2-Aminoethyl)-N-methyl-2-(4-nitrophenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 2-([1,1'-Biphenyl]-4-yl)-N-(2-aminoethyl)-N-methyl-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide N-(2-Aminoethyl)-N-methyl-1-(p-tolyl)-2-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazole-5-carboxamide N-(2-Aminoethyl)-N-methyl-2-(4-(methylsulfonyl)phenyl)-1-(p-tolyl)-1H-benzo[d]imidazole-5-carboxamide 4-(6-(Piperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile 3-(4-Cyanophenyl)-N-(piperidin-4-yl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide 4-(6-(4-Aminopiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile N-(4-Aminobutyl)-3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide N-(2-Aminoethyl)-3-(4-cyanophenyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide 4-(6-(3-Aminopiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile 3-(4-Cyanophenyl)-N-(piperidin-4-ylmethyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide 3-(4-Cyanophenyl)-N-(piperidin-3-yl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide (R)-4-(6-(3-Aminopyrrolidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile 3-(4-Cyanophenyl)-N-(2-(methylamino)ethyl)-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide (R)-4-(6-(2-Methylpiperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile (S)-4-(6-(2-Methylpiperazine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile 4-(6-(4-Methylpiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile 4-(6-(2-Methylpiperidine-1-carbonyl)-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)benzonitrile 3-(4-Cyanophenyl)-N-methyl-2-(p-tolyl)imidazo[1,2-a]pyridine-6-carboxamide.

2. A pharmaceutical composition, characterized in that, It is prepared by mixing the aromatic six-membered ring-fused imidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient with a pharmaceutically acceptable carrier or excipient and formulated into a clinically acceptable dosage form.

3. A method for preparing the aromatic six-membered ring-fused imidazole derivative according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The preparation method of the aromatic six-membered ring-fused imidazole derivative comprises the following steps: Route 1: The starting material A-1 is converted to intermediate A-2 through an esterification reaction. The reaction condition a is esterification under strong acid catalysis, or esterification with an acylating agent, or esterification catalyzed by dicyclohexylcarbodiimide and 4-dimethylaminopyridine, and the reaction temperature is 65 - 85°C; Intermediate A-2 and a substituted aromatic amine or aliphatic amine undergo a nucleophilic substitution reaction to obtain intermediate A-3. The reaction condition b is carried out in the presence of an organic base or an inorganic base as an acid-binding agent. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, and potassium hydroxide; Intermediate A-3 undergoes a reduction reaction to obtain intermediate A-4. The reaction condition c is hydrogenation reduction under a palladium-based catalyst, or reduction using a combination of a reducing metal and an inorganic acid. The combination of the reducing metal and the inorganic acid includes iron powder / ammonium chloride, iron powder / hydrochloric acid, and zinc powder / acetic acid; Intermediate A-4 and a substituted aromatic aldehyde or aliphatic aldehyde undergo a nucleophilic addition dehydration reaction to obtain intermediate A-5. The reaction condition d is carried out under the catalysis of acetic acid, p-toluenesulfonic acid, or a Lewis acid, and the reaction temperature is 80 - 100°C; Intermediate A-5 undergoes a cyclization reaction under iodine catalysis to obtain intermediate A-6. The reaction condition e is carried out under the catalysis of iodine, N-iodosuccinimide, or iodine chloride; Intermediate A-6 is converted to intermediate A-7 through an ester hydrolysis reaction. The reaction condition f is carried out under the catalysis of an organic base or an inorganic base. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, and potassium hydroxide; Intermediate A-7 and a substituted aliphatic amine undergo an acylation reaction to obtain intermediate A-8. The reaction condition g is carried out in the presence of a condensing agent. The condensing agents are 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(benzotriazol-1-yl)-bis(dimethylamino)carbenium hexafluorophosphate, O-(5-chlorobenzotriazol-1-yl)-bis(dimethylamino)carbenium hexafluorophosphate, O-(benzotriazol-1-yl)-bis(dimethylamino)carbenium tetrafluoroborate, O-(N-succinimidyl)-bis(dimethylamino)carbenium tetrafluoroborate, O-(N-endo-5-norbornene-2,3-dicarboximide)-bis(dimethylamino)carbenium tetrafluoroborate, diphenylphosphoryl chloride, diethyl cyanophosphate, diphenylphosphoryl azide, dimethylthiophosphoryl azide, bis(2-oxo-3-azolidinyl)phosphoryl chloride, or 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorin-2,4,6-trioxide; Intermediate A-8 is converted to the target product A-9 through a reaction for removing the Boc protecting group. The reaction condition h is under acid catalysis, heating for 4 - 12 h; The acid is trifluoroacetic acid or hydrochloric acid, and the reaction temperature is 25 - 80°C; Route 2: The reaction conditions and operation methods in Route 2 adopt the corresponding reaction conditions and operation methods in Route 1; Route 3: The starting material B-1 is converted into the intermediate B-2 through an esterification reaction. The reaction condition a is esterification under strong acid catalysis, or esterification with an acylating agent, or esterification catalyzed by dicyclohexylcarbodiimide and 4-dimethylaminopyridine; the reaction solvent is methanol, ethanol or propanol, and the reaction temperature is 80 °C; The intermediate B-2 and the substituted α-halogenated saturated carbonyl compound undergo a cyclization reaction to obtain the intermediate B-3. The reaction condition b is carried out in the presence of an organic base or an inorganic base as an acid-binding agent. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, and potassium hydroxide; the reaction solvents include polar solvents such as methanol, ethanol, propanol, acetone, N,N-dimethylformamide, and dimethyl sulfoxide; The intermediate B-3 is converted into the intermediate B-4 through an iodination reaction. The reaction condition c is iodination catalyzed by iodine, N-iodosuccinimide or iodine monochloride; the reaction solvent is methanol, ethanol, propanol, trifluoroethanol, acetonitrile, acetone, tetrahydrofuran, N,N-dimethylformamide or dimethyl sulfoxide, and the reaction is carried out at 25-60 °C; The intermediate B-4 and the substituted arylboronic acid or aliphatic boronic acid undergo a Suzuki reaction to obtain the intermediate B-5. The reaction condition d is heating under the catalysis of a palladium complex, in the presence of an inorganic base and under an anaerobic condition; the palladium complex is Pd(PPh3)4, PdCl2, PdCl2(dppf), Pd(OAc)2 or Pd(PPh3)2Cl2; the inorganic base is potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate or potassium fluoride, and the reaction solvent is any one or a mixture of two or more of ethanol, 1,4-dioxane, tetrahydrofuran, toluene, N,N-dimethylformamide, dimethyl sulfoxide, water and ethylene glycol dimethyl ether, and the reaction temperature is 80-140 °C; The intermediate B-5 undergoes an ester hydrolysis reaction to obtain the intermediate B-6. The reaction condition e is carried out under the catalysis of an organic base or an inorganic base. The organic bases include triethylamine and N,N-diisopropylethylamine, and the inorganic bases include sodium carbonate, potassium carbonate, cesium carbonate, sodium phosphate, sodium hydroxide, and potassium hydroxide; the reaction solvent is methanol, ethanol, propanol, acetone, N,N-dimethylformamide or dimethyl sulfoxide, and the reaction is carried out at 80 °C; The intermediate B-6 and the substituted aliphatic amine undergo an acylation reaction to obtain the intermediate B-7. The reaction condition f adopts the reaction condition of reaction route g in Route 1; The intermediate B-7 undergoes a reaction to remove the Boc protecting group to obtain the target product B-8. The reaction condition g adopts the reaction condition of reaction route h in Route 1.

4. Use of the aromatic six-membered ring-fused imidazole derivative according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of a lysine-specific demethylase 1 inhibitor.

5. Use of the aromatic six-membered ring-fused imidazole derivative according to claim 1, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 2 in the preparation of an H + , K + -ATPase inhibitor.

6. Use of the aromatic six-membered ring-fused imidazole derivative according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of a drug for inhibiting gastric acid secretion.

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