CN118005608A - Triazole small molecular organic compound for inhibiting mitochondrial oxidative phosphorylation by specific targeting complex I and application thereof - Google Patents

Triazole small molecular organic compound for inhibiting mitochondrial oxidative phosphorylation by specific targeting complex I and application thereof Download PDF

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CN118005608A
CN118005608A CN202211389870.3A CN202211389870A CN118005608A CN 118005608 A CN118005608 A CN 118005608A CN 202211389870 A CN202211389870 A CN 202211389870A CN 118005608 A CN118005608 A CN 118005608A
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methyl
piperazine
carbonyl
trifluoromethyl
benzyl
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陈益华
逄秀凤
孙悦
冯娟娟
何朋
刘明耀
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East China Normal University
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis

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Abstract

The invention discloses a triazole micromolecule organic compound for inhibiting mitochondrial oxidative phosphorylation of a specific targeting complex I, which is characterized by comprising a compound shown in a structure of a formula (I-iii), a pharmaceutical composition containing the compound (I-iii), and pharmaceutically acceptable salts, metabolites or prodrugs. The invention also discloses application of the triazole micromolecular organic compound serving as a mitochondrial oxidative phosphorylation inhibitor in preparing medicines for preventing and/or treating diseases mediated or regulated by oxidative phosphorylation.

Description

Triazole small molecular organic compound for inhibiting mitochondrial oxidative phosphorylation by specific targeting complex I and application thereof
Technical Field
The invention relates to the field of biological medicine, in particular to a novel, efficient and specific triazole micromolecule organic compound for inhibiting mitochondrial oxidative phosphorylation and derivatives thereof, and application of the triazole micromolecule organic compound or a pharmaceutical composition containing the compound in preparation of antitumor drugs.
Background
Tumor metabolic reprogramming includes reprogramming of metabolic pathways such as glucose, amino acids, fatty acids, and nucleotides, with the most classical tumor metabolic reprogramming being the Warburg effect (Warburg effect), which describes that tumor cells undergo glycolysis under both aerobic and anaerobic conditions as compared to normal cells that do glycolysis only under anaerobic conditions, and thus the Warburg effect is also known as aerobic glycolysis. However, in recent years, unlike aerobic glycolysis, it has been found that oxidative phosphorylation (OXPHOS) is mainly relied on in the late stage of development of specific tumor cells, and these specific tumor cells whose oxidative phosphorylation is up-regulated are mainly metastatic, drug resistant, RAS mutated tumors, tumor stem cells, and the like. The important role of oxidative phosphorylation in tumors is constantly being explored, and thus targeting oxidative phosphorylation may be a novel anti-tumor strategy.
In recent years, the development of oxidative phosphorylation inhibitors has been receiving extensive attention, and studies have found that inhibition of oxidative phosphorylation complexes cuts off tumor cell respiratory pathways, inhibits biomass production, and ultimately leads to cancer cell death. Inhibition of oxidative phosphorylation by inhibition of the respiratory chain complex is therefore a viable strategy, especially complex I inhibitors have been most widely studied and clear. Several high affinity complex I (NADH-CoQ reductase complex) inhibitors currently exist, BAY87-2243, IACS-010759, ME-344, mubritinib, DX3-235 and DX3-234. However, at present, some inhibitors still have serious problems of insufficient safety and/or effectiveness, and thus, development of more efficient complex I (NADH-CoQ reductase complex) selective inhibitors is highly demanded. In general, the development of targeted mitochondrial oxidative phosphorylation inhibitors has high clinical demands and development significance.
Disclosure of Invention
The invention aims to design and synthesize a novel and efficient small-molecule triazole inhibitor which specifically targets the mitochondrial complex I to inhibit mitochondrial oxidative phosphorylation, and the small-molecule triazole inhibitor can inhibit mitochondrial respiration by specifically targeting the complex I so as to inhibit the process of NADH (NADH) oxidation to NAD +, thereby achieving excellent antiproliferative activity in tumor cells with high oxidative phosphorylation expression level and drug resistance caused by targeted therapy such as MEK inhibitor and the like, and having wide application prospect.
The invention provides a triazole small molecule organic compound which specifically targets a mitochondrial complex I to inhibit mitochondrial oxidative phosphorylation, wherein the triazole small molecule organic compound comprises a compound shown as a formula (I) or a related analogue or pharmaceutically acceptable salt, metabolite or prodrug of the compound shown as the formula (I); wherein the mitochondrial complex I is an NADH-CoQ reductase complex.
Wherein:
r 1 is selected from any one or more of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoroethoxy, trifluoromethyl, trifluoromethylethoxy, trifluoromethylpropoxy, trifluoromethylamino, C 1～C5 alkyl, C 1～C5 alkoxy;
r 2 is selected from any one or more of the following groups: hydrogen, C 1～C4 alkyl, trifluoromethyl, trifluoroethyl, phenyl, halophenyl, benzyl, phenethyl;
R 3 is selected from any one or more of the following groups: c 1～C6 alkyl, trifluoromethylphenyl, tetrahydropyran, halophenyl, trifluoromethoxyphenyl, C 5～C10 aryl, C 1～C3 alkoxyphenyl, C 2～C6 alkenyl, C 2～C6 alkynyl, C 1～C6 alkoxy, C 1～C6 alkylthio, C 3～C6 cycloalkyl, C 2～C6 cycloalkoxy, C 2～C6 cycloalkylthio, C 2～C6 nitrogen-containing cycloalkyl, 5-10 membered heteroaryl.
In the formula (i) of the present invention, whenIn the 4-position of 1,2, 3-triazole, the structure is shown as (ii):
Wherein:
r 1 is selected from any one or more of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoroethoxy, trifluoromethyl, trifluoromethylethoxy, trifluoromethylpropoxy, trifluoromethylamino, C 1～C5 alkyl, C 1～C5 alkoxy;
r 2 is selected from any one or more of the following groups: hydrogen, C 1～C4 alkyl, trifluoromethyl, trifluoroethyl, phenyl, halophenyl, benzyl, phenethyl;
R 3 is selected from any one or more of the following groups: c 1～C6 alkyl, trifluoromethylphenyl, tetrahydropyran, halophenyl, trifluoromethoxyphenyl, C 5～C10 aryl, C 1～C3 alkoxyphenyl, C 2～C6 alkenyl, C 2～C6 alkynyl, C 1～C6 alkoxy, C 1～C6 alkylthio, C 3～C6 cycloalkyl, C 2～C6 cycloalkoxy, C 2～C6 cycloalkylthio, C 2～C6 nitrogen-containing cycloalkyl, 5-10 membered heteroaryl.
In the formula (ii), when R 3 is aryl, the structure is shown as (iii):
Wherein:
r 1 is selected from any one or more of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoroethoxy, trifluoromethyl, trifluoromethylethoxy, trifluoromethylpropoxy, trifluoromethylamino, C 1～C5 alkyl, C 1～C5 alkoxy;
r 2 is selected from any one or more of the following groups: hydrogen, C 1～C4 alkyl, trifluoromethyl, trifluoroethyl, phenyl, halophenyl, benzyl, phenethyl;
R 4 is selected from any one or more of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoromethyl, trifluoromethylamino, C 1～C3 alkyl, C 1～C3 alkoxy;
Selected from 5-to 8-membered aromatic rings or aromatic heterocycles, taken from any one of the following aromatic rings: benzene ring, pyridine, pyridazine, pyrimidine, pyrazine, thiophene, thiazole, isothiazole, pyrrole, pyrazole, imidazole, oxazole, oxadiazole, 1,2, 3-triazole, 1,2, 4-triazole, naphthalene ring, phenanthrene ring, quinoline, isoquinoline, indole, benzodiazole, benzotriazol, purine.
The triazole small molecule organic compound for specifically targeting the mitochondrial complex I to inhibit mitochondrial oxidative phosphorylation comprises any one of the following compounds, or pharmaceutically acceptable salts, metabolites or prodrugs thereof:
5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
N-ethyl-5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) pyridine amide
N-isobutyl-5- (5-methyl-4- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazole-1-methyl) pyridine amide
5- ((5-Methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (tetrahydro-2H-pyran-4-yl) picolinamide
5- ((5-Methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (2- (trifluoromethyl) phenyl) pyridine amide
5- ((5-Methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (3- (trifluoromethyl) phenyl) pyridine amide
N- (4-fluorophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide
N- (4-chlorophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide
N- (4-bromophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide
5- ((5-Methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethoxy) phenyl) pyridinecarboxamide
5- ((5-Methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (naphthalen-2-yl) picolinamide
5- ((5-Methyl-4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5-Methyl-4- (4- (3- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5-Methyl-4- (4- (4- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5-Methyl-4- (4- (4- (trifluoromethoxy) benzoyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((4- (4- (4- (2, 2-Trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5-Ethyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((4- (4- (4- (2, 2-Trifluoroethoxy) benzyl) piperazine-1-carbonyl) -5- (trifluoromethyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5-Phenyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((4-Methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethoxy) phenyl) pyridinecarboxamide
5- ((4-Methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N-phenylpyridine amide
N- (4-methoxyphenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide
N- (4- (tert-butyl) phenyl) -5- (4-methyl-5- (4- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazole-1-methyl) pyridine amide
5- (4-Methyl-5- (4- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (p-tolyl) pyridine amide
N- (4-methoxy-3- (trifluoromethyl) phenyl) -5- (4-methyl-5- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) picolinamide
N- (4-fluorophenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide
N- (4-bromophenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide
5- ((4-Methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridinecarboxamide
5- ((4-Methyl-5- (4- (4- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((4-Methyl-5- (4- (4-methylbenzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5- (4- (4-Chloro-3- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5- (4- (4-Isopropylbenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5- (4- (4- (Tert-butyl) benzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5- (4- (4-Chlorobenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- (5- (4- (3-Bromo-4-fluorobenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl-N- (4-trifluoromethyl) phenyl) picolinamide
5- ((5- (4-Benzylpiperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazolyl) methyl) -N- (4-trifluoromethyl) phenylpyridine amide
5- ((4-Methyl-5- (4- (methylsulfonyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((5- (4- (4- (2, 2-Trifluoroethoxy) benzyl) piperazine-1-carbonyl) -4- (trifluoromethyl) -1H-1,2, 3-triazol-1-yl) methyl) -N-4- (trifluoromethyl) phenyl) pyridine amide
5- ((4-Ethyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
5- ((4-Phenyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide
The term "C 1～C6 alkyl" in the present invention refers to straight or branched chain alkyl groups having 1 to 6 carbon atoms and includes, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl and the like.
The term "C 2～C6 alkenyl" in the present invention refers to a straight or branched alkenyl group having at least one double bond of 2 to 6 carbon atoms, including without limitation ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, butadienyl, pentenyl, pentadienyl, hexenyl, hexadienyl, tricenyl and the like.
The term "C 2～C6 alkynyl" according to the present invention refers to a straight or branched alkynyl group containing at least one triple bond having 2 to 6 carbon atoms, including without limitation ethynyl, propynyl, isopropoxynyl, butynyl, isobutynyl, butadiynyl, pentynyl, glutaryl, alkynyl-ended, diynyl and the like.
The term "C 1～C6 alkoxy" in the present invention refers to a straight or branched chain alkoxy group containing at least one oxygen atom having 1 to 6 carbon atoms, including without limitation methoxy, ethoxy, propoxy, ethoxymethoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, 2-methyl (2-butoxy, 3-methyl (1-butoxy, 2-methyl (1-butoxy, n-hexyloxy, etc.).
The term "C 3～C6 cycloalkyl" in the present invention refers to cycloalkanes having 3 to 6 carbon atoms and includes, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
The term "C 2～C6 cycloalkoxy" according to the present invention refers to a cycloalkoxy group having at least one oxygen atom with 2 to 6 carbon atoms, including without limitation ethylene oxide, propylene oxide, tetrahydrofuran, tetrahydropyranyl, and the like.
The term "C 2～C6 nitrogen-containing cycloalkyl" according to the present invention refers to cycloalkyl groups having 2 to 6 carbon atoms containing at least one nitrogen atom, including, without limitation, tetrahydropyrrolyl, morpholinyl, and the like.
The term "C 5～C10 aromatic group" in the present invention refers to an aromatic ring group having 5 to 10 carbon atoms which does not contain a heteroatom in the ring, and includes, without limitation, phenyl, naphthyl, phenanthryl, and the like.
The term "5-10 membered heteroaromatic group" of the present invention has a 5-to 10-membered aromatic ring of one or more heteroatoms selected from nitrogen, oxygen or sulfur, including without limitation pyridyl, pyrimidinyl, thiazolyl, isothiazolyl, furyl, thienyl, pyrrolyl, indolyl, quinolinyl, isoquinolinyl, and the like.
The term "halogen" in the present invention means fluorine, chlorine, bromine, or iodine.
The term "pharmaceutically acceptable salt" of the present invention refers to salts of the compounds of the present invention prepared from the compounds of the present invention which have the specified substituents found herein and relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting neutral forms of such compounds with a sufficient amount of a base in pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, amine, organic amine or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of an acid in pure solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts including acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also included are salts of amino acids (e.g., arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the invention contain both acidic and basic functionalities and can be converted to either base or acid addition salts.
In addition to salt forms, the compounds provided herein exist in prodrug forms. Prodrugs of the compounds described herein are susceptible to chemical changes under physiological conditions that convert to the compounds of the invention. Any compound that can be converted in vivo to provide a biologically active substance (i.e., a compound of formulas i-iii) is a prodrug within the scope and spirit of the invention. For example, compounds containing carboxyl groups can form physiologically hydrolyzable esters that act as prodrugs by hydrolyzing in vivo to give the compounds of formula (i-iii) themselves.
Certain compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereomers, geometric isomers and individual isomers are included within the scope of the present invention, including cis and trans isomers, (-) -and (+) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, and racemic and other mixtures thereof, and all such isomers and mixtures thereof are included within the scope of the present invention.
The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more of the atoms comprising the compounds. For example, compounds may be labeled with isotopes, such as deuterium (2 H), tritium (3 H), iodine-125 (125 I) or C-14 (14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.
The term "excipient" according to the present invention generally refers to the carrier, diluent and/or medium required to formulate an effective pharmaceutical composition.
For a drug or pharmacologically active agent, the term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of the drug or agent that is non-toxic but achieves the desired effect. For the purposes of the oral dosage form of the present invention, an "effective amount" of one active agent in a composition refers to that amount of the other active agent in the composition which is required to achieve the desired effect when combined. Determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, and also on the particular active substance, a suitable effective amount in an individual case can be determined by one skilled in the art according to routine experimentation.
The term "active ingredient", "therapeutic agent", "active substance" or "active agent" as used herein refers to a chemical entity that is effective in treating a disorder, disease or condition of interest.
When any variable (e.g., R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0 to 2R, the group may optionally be substituted with up to two R's, and R's in each case are independent options. Furthermore, combinations of substituents and/or variants thereof are only permissible if such combinations result in stable compounds.
The term "comprising" in this application is an open-ended expression, i.e., including what is indicated in the present application, but not excluding other aspects.
The invention also provides a pharmaceutical composition which contains the triazole small molecular organic compound or an analogue or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, wherein the triazole small molecular organic compound is used for specifically targeting the mitochondrial complex I to inhibit mitochondrial oxidative phosphorylation.
The pharmaceutical composition further comprises a second agent.
The second medicament is used for preparing a medicament for preventing and/or treating cancers.
The pharmaceutical compositions contain a therapeutic dose of the aforementioned compounds.
"Pharmaceutical composition" means a mixture containing one or more of the compounds of the present invention or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, or other components such as physiological saline/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration to organisms, facilitate the absorption of active ingredients and thus exert biological activity.
According to a specific embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or combination thereof.
According to a specific embodiment of the present invention, the pharmaceutical composition is in the form of a tablet, capsule, injection, powder for injection, powder, syrup, solution, suspension or aerosol. Thereby significantly improving the applicability of the pharmaceutical composition. And the pharmaceutical compositions of the above embodiments of the present invention may be present in a suitable solid or liquid carrier or diluent and in a suitable sterile container for injection or instillation.
The various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional preparation methods in the pharmaceutical field. The compounds and pharmaceutical compositions of the present invention may be administered to mammals, including humans and animals, by oral, nasal, dermal, pulmonary or gastrointestinal routes. Regardless of the method of administration, the optimal dosage for an individual will depend on the particular treatment regimen. Typically starting from a small dose, the dose is gradually increased until the most appropriate dose is found. The most preferred route of administration is oral or transdermal.
According to a specific example of the present invention, the compounds of formulas i-iii according to the present invention have an inhibitory effect on mitochondrial oxidative phosphorylation.
According to the examples of the invention, the compounds shown in the formulas i-iii have obvious inhibition effects on proliferation, growth, infiltration, clone formation and metastasis of various cancer cells, have obvious promotion effects on apoptosis of various cancer cells, and can prolong the survival time of tumor patients.
According to an embodiment of the present invention, the combination of the second agent with the compounds of formulae i to iii makes the pharmaceutical composition more effective for inhibiting mitochondrial oxidative phosphorylation and for treating or preventing cancer, renal fibrosis, pulmonary fibrosis, rheumatoid arthritis, psoriasis, lupus erythematosus, inflammatory lung diseases, inflammatory bowel diseases, and the like.
The invention also provides a mitochondrial oxidative phosphorylation inhibitor, which comprises the triazole small molecular organic compound and pharmaceutically acceptable salts, metabolites or prodrugs for specifically targeting the mitochondrial complex I to inhibit mitochondrial oxidative phosphorylation.
The invention also provides application of the mitochondrial oxidative phosphorylation inhibitor in treatment of diseases related to mitochondrial oxidative phosphorylation dysfunction, such as tumors relying on oxidative phosphorylation metabolism, hypoxic tumors, tumors resistant to targeted therapy or other therapies, tumor stem cells and the like.
The invention also provides application of the triazole small molecular organic compound for inhibiting mitochondrial oxidative phosphorylation of the specific targeted mitochondrial complex I, pharmaceutically acceptable salts, metabolites or prodrugs or the pharmaceutical composition in preparation of oxidative phosphorylation inhibitors.
The invention also provides application of the triazole small molecular organic compound for inhibiting mitochondrial oxidative phosphorylation of the specific targeted mitochondrial complex I, or pharmaceutically acceptable salt, metabolite or prodrug or the pharmaceutical composition in preparation of an inhibitor for inhibiting oxidative phosphorylation of tumor cells with high oxidative phosphorylation level and inhibiting oxidative phosphorylation by specifically inhibiting respiratory chain complex I.
The medicament of the invention can be effectively used as an oxidative phosphorylation inhibitor for treating diseases related to mitochondrial oxidative phosphorylation activation, wherein the diseases are taken from malignant tumors, such as lung cancer, pancreatic cancer, colorectal cancer, ovarian cancer, breast cancer, acute myeloid leukemia and other tumors which depend on oxidative phosphorylation metabolism and oxidative phosphorylation level up-regulation; the metastasis of the tumor and cancer.
The invention further provides a method for preventing and/or treating a disease associated with mitochondrial oxidative phosphorylation activation comprising administering to a tumor patient an effective amount of a compound or pharmaceutical composition or mitochondrial oxidative phosphorylation inhibitor as described above.
The invention provides a preparation method of the compound shown in the formulas i-iii, wherein the preparation method of the compound is selected from any one of the following synthetic methods, and R 1、R2 and R 3 are defined as the invention.
Wherein, the synthesis method 1 is as follows:
The synthesis method 2 is as follows:
The invention has the beneficial effects that: the triazole micromolecular organic compound or the pharmaceutical composition for targeting mitochondrial oxidative phosphorylation can be used as an oxidative phosphorylation inhibitor, is used for preventing and/or treating diseases caused or regulated by mitochondrial oxidative phosphorylation, and has good clinical application and medical application.
Drawings
FIG. 1 shows the inhibition of mitochondrial respiration rate and glycolysis by representative compound SY1-001, wherein A is the inhibition of cellular oxygen consumption, basal respiration rate, maximum respiration rate and respiration potential by representative compound SY1-001 in high-expression oxidative phosphorylated lung cancer cell H460, B is the inhibition of cellular oxygen consumption, basal respiration rate, maximum respiration rate and respiration potential by representative compound SY1-001 in high-expression oxidative phosphorylated lung cancer cell H441, C is the inhibition of cellular oxygen consumption, basal respiration rate, maximum respiration rate and respiration potential by representative compound SY1-001 in low-expression oxidative phosphorylated lung cancer cell H358, and D is the inhibition of extracellular acidification rate by representative compound SY1-001 in lung cancer cells H460, H441 and H358.
FIG. 2 shows the inhibition of mitochondrial respiration rate and glycolysis by representative compounds SY1-001, etc.
FIG. 3 shows the promotion of apoptosis of high-expression oxidative phosphorylation tumor cells by representative compounds SY1-001 in vitro.
FIG. 4 shows the sensitization of the MEK inhibitor trametinib in vitro by representative compounds SY 1-001.
FIG. 5 shows the antitumor effect of representative compound SY1-001 alone or in combination with trametinib in vivo, wherein A is the inhibition of the growth of the lung cancer xenogenic tumor volume by representative compound SY1-001 alone or in combination with trametinib, B is the statistics of the area under the tumor volume of the inhibition of the lung cancer xenogenic tumor by SY1-001 alone or in combination with trametinib, C is the effect of representative compound SY1-001 alone or in combination with trametinib on the weight of the lung cancer xenogenic tumor, and D is the effect of representative compound SY1-001 alone or in combination with trametinib on the weight of mice.
Detailed Description
The present invention will be described in further detail with reference to the following specific examples and drawings. The procedures, conditions, experimental methods, etc. for carrying out the present invention are common knowledge and common knowledge in the art, except for the following specific references, and the present invention is not particularly limited.
Example one specific Synthesis of Compounds of the invention
Example 1-1,5 Synthesis of- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-001).
5- (Methoxycarbonyl) -2-pyridinecarboxylic acid (5.0 g,27.60 mmol), para-trifluoromethylaniline (5.3 g,33.12 mmol), HATU (12.5 g,33.12 mmol) were weighed in a 100mL round bottom flask, DIEA (14 mL,82.80 mmol) was added dropwise, stirred at ambient temperature for 2h, water was added and suction filtered, the resulting solid was dried over anhydrous sodium sulfate, and the solid was slurried with ethyl acetate to give intermediate 6- ((4- (trifluoromethyl) phenyl) carbamoyl) nicotinic acid methyl ester 4.0g, yield 78%; then, intermediate 6- ((4- (trifluoromethyl) phenyl) carbamoyl) methylnicotinate (4.0 g,13.73 mmol) was dissolved in 20mL THF solution, air was replaced with nitrogen, and the mixture was placed in a low temperature reactor and stirred at-20℃for three to five minutes, 2eq LiAlH 4 (27 mL,27.46 mmol) was added dropwise to the reaction system, reacted at-20℃for 1-2 hours, ice water was added dropwise to quench LiAlH 4, the funnel pad of the sand was suction-filtered, ethyl acetate was used to rinse LiAlH 4 colloid, the filtrate was dried by rotary evaporation, and purified by column chromatography to give intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 2.4g in 59% yield.
Intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (2.4 g,8.25 mmol) was dissolved in 20mL DCM, stirred in ice bath, PBr 3 (0.6 mL,6.19 mmol) was added dropwise under ice bath, reacted for 1-2h under ice bath, PBr 3 was quenched dropwise with ice water, extracted, dried, column chromatographed to give intermediate 5- (bromomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 1.4g, yield 50%; intermediate 5- (bromomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (1.4 g,3.89 mmol) was weighed out, naN 3 (304 mg,4.67 mmol) was dissolved in 4mL DMF and reacted at room temperature for 2h, water was added, EA extraction, back extraction, drying to give intermediate 5- (azidomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 476mg, yield 90%.
Intermediate 5- (azidomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (470 mg,1.48 mmol) and ethyl 2-butynoate (1.0 g,8.92 mmol) were dissolved in 12mL toluene solution, heated to 120 ℃, reacted overnight, the toluene solution was dried by spin-drying, extracted with water, dried, purified by column chromatography to give intermediate 5-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-4-carboxylic acid ethyl ester 131mg, yield 21%; intermediate 5-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-4-carboxylic acid ethyl ester (131 mg,0.31 mmol) was then dissolved in 20mL of CH 3OH:H2 o=4:1 mixed solvent, lioh. 2 O (52 mg,1.24 mmol) was added, heated and stirred at 60 ℃ for 1H, dried methanol was added, concentrated hydrochloric acid solution was added to adjust PH to 1-2, ea extraction, and the solid was dried to give intermediate 5-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-4-carboxylic acid 86mg, 69% yield.
Intermediate 5-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-4-carboxylic acid (86 mg,0.21 mmol), 1- (4- (2, 2-trifluoroethoxy) benzyl) piperazine hydrochloride (79 mg,0.25 mmol), HATU (97 mg,0.25 mmol) were weighed out in DMF solution, DIEA (0.14 ml,0.84 mmol) was added and reacted overnight at room temperature to give the target product 5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-001) 120mg, yield, purified by conventional work-up and column chromatography ：87％.1H NMR(600MHz,Chloroform-d)δ10.08(s,1H),8.54(d,J＝2.2Hz,1H),8.30(d,J＝8.1Hz,1H),7.90(d,J＝8.4Hz,2H),7.77(dd,J＝8.1,2.2Hz,1H),7.65(d,J＝8.4Hz,2H),7.36–7.25(m,2H),6.96–6.89(m,2H),5.62(s,2H),4.36(q,J＝8.1Hz,2H),4.14(s,2H),3.80(s,2H),3.54(s,2H),2.56(tt,J＝16.1,8.0Hz,4H),2.49(s,3H).
Example II, inhibitory Activity of the Compounds of the invention against mitochondrial respiratory chain complexes I-IV (Complex I-IV refers to NADH-CoQ reductase complex, succinate dehydrogenase, UQH-cytochrome c oxidoreductase and cytochrome c oxidase, respectively) and NAD +/NADH
2-1, Technical method:
(1) Electron flow experiment
Electron flow experiments were performed using a Seahorse instrument to analyze electron transfer chain function and its relationship to mitochondrial oxidative phosphorylation. Cells were digested and collected, 2×10 5 cells were plated in XF96 cell culture plates (four background wells (A1, a12, H1, H12) without plating cells). Meanwhile, the detection probes were placed in hydration solution and incubated overnight in a CO 2 -free incubator. After the cells are attached, the compounds SY 1-001-SY 1-040 prepared in the embodiment of the invention are respectively added for treatment, and 3-6 compound holes are arranged for each treatment. After the compounds SY 1-001-SY 1-040 prepared in the embodiment of the invention are treated for 6 hours respectively, the cell culture medium is discarded, 200 mu L of detection liquid is added into each hole to wash the cells, and the process is repeated twice. 180. Mu.L of detection solution was added to each well last time (including four background wells). To the hydrated test plates a, b, c and d, 20. Mu.L of 10. Mu.M rotenone, 22. Mu.L of 10mM succinic acid, 25. Mu.L of 4. Mu.M oligomycin A and 28. Mu.L of a mixture of 10mM sodium ascorbate and 100. Mu.M tetramethylp-phenylenediamine were added, respectively. And setting instrument parameters, adjusting a program and starting a test. Experimental data were analyzed using Wave software and report generator.
(2) Determination of NAD +/NADH ratio
Cells are evenly inoculated into a 6-well plate with proper density, and after the cells are completely attached, the compounds SY 1-001-SY 1-040 prepared by the embodiment of the invention are added into each well for treatment. After 24h of drug action, the NAD +/NADH ratio in the cells was determined using the NAD +/NADH detection kit. The specific operation flow is as follows: cells were lysed and centrifuged at 12000rpm for 20min at 4℃and the supernatant was collected as the sample to be tested. Sucking 20 mu L of NADH standard substances with different concentrations into a 96-well plate for preparing a standard curve; sucking 20 mu L of the sample to be detected into a 96-well plate for measuring the total amount of NAD + and NADH in the sample; sucking 50-100 mu L of a sample to be detected into a centrifuge tube, carrying out water bath at 60 ℃ for 30min, centrifuging, taking 20 mu L of supernatant into a 96-well plate, and measuring the concentration of NADH in the sample; and adding 90 mu L of ethanol dehydrogenase working solution into the blank control hole, the standard substance hole and the sample hole respectively, fully and uniformly mixing, and incubating for 10min at 37 ℃ in a dark place. After the incubation, 10. Mu.L of color development solution was added to each well, and incubated at 37℃for 30min in the dark, at which time orange formazan was formed, and the absorbance of the sample was measured at 450 nm. The amount of NAD + in the sample and the NAD +/NADH ratio were calculated according to the following calculation formula: NAD +＝NADtotal-NADH、NAD+/NADH＝(NADtotal -NADH)/NADH).
TABLE 1 inhibitory Activity of the inventive Compounds SY1-001 to SY1-040 against mitochondrial respiratory chain complexes I-IV and NAD +/NADH at a concentration of 1. Mu.M
2-2 Conclusion
The experimental results are shown in table 1, and after screening through electron flow experiments and NAD +/NADH experiments, the compounds SY1-001 to SY1-040 prepared in the embodiment of the invention can selectively inhibit the activity of the respiratory chain complex I to different degrees (wherein <0 in the table indicates no inhibitory activity), and have different degrees of inhibitory activity on NAD +/NADH, wherein most of the compounds can have 40% or more of the inhibitory rate on the respiratory chain complex I (NADH-CoQ reductase complex), and the compounds can inhibit oxidative phosphorylation through specific targeting complex I (NADH-CoQ reductase complex); in addition, the inhibition rate of most compounds to NAD +/NADH can reach 50%, and further the compounds can inhibit the process of oxidizing NADH into NAD + through specific targeting complex I (NADH-CoQ reductase complex). Such as SY1-001, SY1-005, SY1-007, SY1-008, SY1-009, SY1-010, SY1-015, SY1-017, SY1-024, SY1-027, SY1-028, SY1-035 and the like having an inhibition ratio to complex I and an NAD +/NADH inhibition ratio of more than 60%; compounds such as SY1-006、SY1-011、SY1-013、SY1-014、SY1-014、SY1-015、SY1-016、SY1-018、SY1-020、SY1-022、SY1-030、SY1-033、SY1-034、SY1-037、SY1-038 with inhibition rate on complex I and NAD +/NADH inhibition rate of 40% -60%.
Example III antiproliferative Activity of the Compounds of the invention on high-expression oxidative phosphorylation and Low-expression oxidative phosphorylation tumor cells
3-1, Technical method:
(1) SRB method for detecting tumor cell proliferation
The harvested cells were digested and seeded in 96-well plates at a lung cancer cell and pancreatic cancer cell density of 3,000-5,000 per well as described in table 2 below. After the cells are completely adhered, the compounds SY 1-001-SY 1-40 prepared in the published example with different concentrations are added for treatment, and 3 compound holes are arranged in each concentration. After 48h of the respective actions of SY1-001 to SY1-040 prepared in the examples of the present invention, cell proliferation was detected by a Sulforhodamine B (SRB) colorimetric method and the half maximal inhibitory concentration (IC 50) of the drug was calculated using GRAPHPAD PRISM 8.0.0 software.
TABLE 2 antiproliferative activity of the compounds SY1-001 to SY1-040 prepared in the examples of the invention in tumor cells with high and low expression of oxidative phosphorylation
3-2 Conclusion
Table 2 shows that the compounds SY1-001 to SY1-040 prepared in the embodiment of the invention have an unnecessary degree of inhibition effect on oxidative phosphorylation highly expressed lung cancer cells, pancreatic cancer cells, colorectal cancer cells, ovarian cancer cells, breast cancer cells and acute myeloid leukemia cells, and most of the compounds show excellent anti-tumor cell proliferation effect in the oxidative phosphorylation highly expressed cancer cell lines; compounds such as SY1-002、SY1-005、SY1-012、SY1-014、SY1-018、SY1-019、SY1-020、SY1-024、SY1-027、SY1-030、SY1-031、SY1-034、SY1-035、SY1-036 having an IC 50 of less than 100nM inhibitory activity; compounds having an inhibitory activity of less than 50nM in IC 50, such as SY1-003, SY1-006, SY1-009, SY1-013, SY1-015, SY1-023, SY1-029, and SY 1-038; compounds having an IC 50 of less than 10nM inhibitory activity, such as SY1-001, SY1-007, SY1-010, SY1-014, SY1-021, SY1-025, SY1-033, SY1-037, and SY 1-039. These results demonstrate that the compounds of the present invention have good antiproliferative effects on different types of high-expression oxidative phosphorylation tumors; in addition, table 2 also shows the inhibitory activity of the compounds of the present invention in tumor cells or normal cells under-expressing oxidative phosphorylation, and the results show that none of the compounds has significant inhibitory activity, indicating the selectivity and specificity of the compounds of the present invention for tumor cells dependent on oxidative phosphorylation metabolism.
Example IV inhibition of mitochondrial respiratory Rate and glycolysis by representative Compounds SY1-001, et al
4-1, The technical method comprises the following steps:
(1) Determination of cell oxygen consumption Rate (oxygen consumption rate, OCR)
The OCR of the adherent high-expression oxidative phosphorylated lung cancer cells H460 and H441 and the low-expression oxidative phosphorylated lung cancer cells H358, as well as lung cancer tumor mass cells, were detected using an XF96 extracellular flux analyzer. For adherent cells, cells were digested and collected, and cells were seeded in XF96 cell culture plates (four background wells (A1, a12, H1, H12) without seeding cells). After the cells are attached, the representative compound SY1-001 is added for treatment, and 3-6 compound holes are arranged for each treatment. After the drug action is finished, the cell culture medium is discarded, 200 mu L of XF cell mitochondrial pressure test detection liquid is added into each hole to wash the cells, and the cell washing is repeated twice. 180 mu L of XF cell mitochondrial pressure test detection liquid is added into each hole (comprising four background holes) for the last time, and the mixture is placed in a carbon dioxide-free incubator at 37 ℃ for incubation for 1h. To hydrated test plates a, b, c were added 20. Mu.L of 10. Mu.M oligomycin (oligomycin, oligo), 22. Mu.L of 10. Mu.M carbonyl cyanide 4-phenylhydrazone (carbonyl cyanide-trifluoromethoxyphenyihydrazone, FCCP) and 25. Mu.L of 5. Mu.M rotenone/oligomycin A (rotenone/ANTIMYCIN A, rot), respectively. And setting an experimental program, and detecting. (Note that the present disclosure is similar to SY1-001 in other technical methods for detecting representative compounds such as SY1-002 to SY 1-0040)
For lung cancer tumor mass cells, tumor tissues are separated from tumor-bearing mice after the end of the administration treatment, the tumor tissues are sheared by surgical scissors in a six-hole plate added with 1mL of prepared tumor tissue digestion solution, and the tumor tissues are placed in a 37 ℃ incubator for digestion for 40min and then filtered by a filter screen. Collecting suspension, centrifuging, removing supernatant, adding 3mL erythrocyte lysate, lysing for 10min, stopping lysing red in complete culture medium, and filtering with filter screen to obtain single cell suspension. After centrifugation, the supernatant was removed, and the sample was resuspended and rinsed once with XF cell mitochondrial pressure test assay. After treatment, the cells are counted and diluted to a suitable density, typically 10 8 cells/mL, using the XF cell mitochondrial pressure test assay. 80. Mu.L of the suspension was inoculated into an XF96 Cell culture plate previously plated with Cell TAK, and the cells were not inoculated into four background wells (A1, A12, H1, H12), centrifuged at 100g for 10min, and the detection was completed in 1H as described above. Experimental data were analyzed using Wave 2.4 software and report generator. (Note that the present disclosure is similar to SY1-001 in other technical methods for detecting representative compounds such as SY1-002 to SY 1-0040)
(2) Determination of extracellular acidification Rate (extracellular acidification rate, ECAR)
ECAR of adherent tumor cells was detected using XF96 extracellular flux analyzer. Cells were inoculated into XF96 cell culture plates according to the above method, and after the cells were attached, the representative compound SY1-001 of the present invention was added to perform treatments, and 3-6 wells were set for each treatment. After the drug action is finished, the cell culture medium is discarded, 200 mu L of XF cell glycolysis pressure test detection liquid is added into each hole to wash the cells, and the steps are repeated twice. 180 mu L of XF cell glycolysis pressure test detection liquid is added into each hole for the last time, and the mixture is placed in a carbon dioxide-free incubator at 37 ℃ for incubation for 1h. mu.L of 100mM glucose (glucose), 22. Mu.L of 20. Mu.M oligomycin A (oligomycin, oligo) and 25. Mu.L of 500mM 2-deoxyglucose (2-Deoxy-D-glucose, 2-DG) were added to the hydrated wells of the test plates a, b, c, respectively. And setting an experimental program, and detecting. Experimental data were analyzed using Wave 2.4 software and report generator. (Note that the present disclosure is similar to SY1-001 in other technical methods for detecting representative compounds such as SY1-002 to SY 1-0040)
4-2 Conclusion
In order to further examine the selective inhibition of mitochondrial oxidative phosphorylation of tumor cells by the compounds of the present invention, the present invention selects SY1-003、SY1-006、SY1-009、SY1-013、SY1-015、SY1-023、SY1-029、SY1-038、SY1-001、SY1-007、SY1-010、SY1-014、SY1-021、SY1-025、SY1-033、SY1-037、SY1-039 and other compounds having antiproliferative activity (IC 50) below 50nM in tumor cells with high oxidative phosphorylation levels to further examine the inhibition of cell Oxygen Consumption Rate (OCR) and extracellular phosphorylation rate (ECAR) in tumor cells with high oxidative phosphorylation levels, and finds that the above selected compounds have the effect of selectively inhibiting oxidative phosphorylation of tumor cells. Among these, the present invention mainly shows the experimental results of representative compounds SY 1-001. As shown in fig. 1, the representative compound SY1-001 was able to inhibit the cell Oxygen Consumption Rate (OCR), mitochondrial basal respiration rate, maximum mitochondrial respiration rate and respiration potential of H460 and H441 lung cancer cell lines highly expressing oxidative phosphorylation at 200nM concentration (fig. 1A and 1B), while no significant inhibition was observed on the cell oxygen consumption rate of H358 lung cancer cell lines low expressing oxidative phosphorylation (fig. 1C), indicating that the representative compound SY1-001 selectively inhibited oxidative phosphorylation function of highly expressing oxidative phosphorylated tumor cells. In addition, SY1-001 has no obvious inhibition effect on the extracellular acidification rate (ECAR) of high-expression and low-expression oxidative phosphorylation lung cancer cell lines (figure 1D), namely, the SY1-001 has no obvious inhibition effect on glycolysis of tumor cells, and further shows that SY1-001 and other compounds can specifically inhibit oxidative phosphorylation metabolic pathways of the tumor cells.
Example five, inhibition of cloning of highly expressed oxidative phosphorylation tumor cells by representative Compounds SY1-001 in vitro
5-1, Technical method: 2D cloning experiments
H460, H441, A549/TR and H358 cells were plated into 12-well plates at a density of 3000-5000 per well. After adherence, treatment with various concentrations of the representative compound SY1-001 of the present invention was carried out for each well, and the culture was continued for 7-10 days. When apparent cell clones were seen, the medium was discarded, and 250 μl of 4% Paraformaldehyde (PFA) was added to each well and fixed at room temperature in the dark for 15min. The fixative was discarded and 250. Mu.L of 0.1% crystal violet per well was added for 8min. And finally, washing with pure water for 3 times, airing at room temperature and photographing. For clone quantification, crystal violet was dissolved using an appropriate volume of 10% acetic acid solution, 100 μl was placed in a 96-well plate, three wells were set, and absorbance was measured at 595 nm. (it should be noted that the technical methods of the present disclosure for the detection of other published compounds such as SY1-002 to SY1-0040 are similar to SY 1-001) experimental data were analyzed using GRAPHPAD PRISM 8.0.0 software.
5-2 Conclusion
In order to further detect the antiproliferative effect of the compound on the tumor cells with high expression oxidative phosphorylation, the invention selects the compounds with antiproliferative activity (IC 50) lower than 50nM SY1-003、SY1-006、SY1-009、SY1-013、SY1-015、SY1-023、SY1-029、SY1-038、SY1-001、SY1-007、SY1-010、SY1-014、SY1-021、SY1-025、SY1-033、SY1-037、SY1-039 and the like in the tumor cells with high oxidative phosphorylation to further detect the influence on the clone formation of the tumor cells with high oxidative phosphorylation in vitro, and finds that the selected compounds have the effect of resisting the clone formation of the tumor cells with high oxidative phosphorylation. Among these, the present invention mainly shows the experimental results of representative compounds SY 1-001. As shown in FIG. 2, the representative compound SY1-001 can well inhibit the clone formation of high-expression oxidative phosphorylation lung cancer cell strains H460, H441 and A549/TR at the concentration of 100nM in vitro, and has no obvious inhibition effect on the clone formation of low-expression oxidative phosphorylation lung cancer cell strain H358, which indicates the specificity of the compound SY1-001 and the like prepared by the embodiment of the invention on tumor cells which depend on oxidative phosphorylation metabolism.
Example six in vitro promotion of apoptosis of high-expression oxidative phosphorylation tumor cells by representative Compounds SY1-001
6-1, Technical method: apoptosis detection
H460 and H441 cells were seeded in 6 cm diameter dishes and cultured overnight. The next day, SY1-001 prepared in the example of the invention was treated for 48h with 30 nM. After the drug treatment is completed, the cells are digested and collected. mu.L of 1 Xbinding Buffer, 3.5. Mu.L of PI and 3.5. Mu.L of Annexin V were added to each sample and incubated for 15min in the dark. Cells were filtered and transferred to a flow tube and analyzed by flow cytometry on-machine. (other representative compounds of the present disclosure, such as SY 1-002-SY 1-0040, are similar to SY 1-001)
6-2 Conclusion
In order to further detect the pro-apoptosis effect of the compounds of the invention on the tumor cells with high expression oxidative phosphorylation, the invention selects the compounds with anti-proliferation activity (IC 50) lower than 50nM SY1-003、SY1-006、SY1-009、SY1-013、SY1-015、SY1-023、SY1-029、SY1-038、SY1-001、SY1-007、SY1-010、SY1-014、SY1-021、SY1-025、SY1-033、SY1-037、SY1-039 and the like in the tumor cells with high oxidative phosphorylation to further detect the pro-apoptosis effect on the tumor cells with high and low oxidative phosphorylation, and discovers that the selected compounds have the effect of promoting the apoptosis of the tumor cells with high expression oxidative phosphorylation. Among these, the present invention mainly shows the experimental results of representative compounds SY 1-001. As shown in the experimental result in figure 3, compared with a control group, the compound SY1-001 can obviously induce 35% -45% of lung cancer cells to apoptosis in vitro, which shows that the compound SY1-001 and the like can obviously promote the apoptosis of high-expression oxidative phosphorylation tumor cells.
Example seven sensitization of the representative Compound SY1-001 to the MEK inhibitor trametinib in vitro
7-1, Technical method: SRB method for detecting tumor cell proliferation (its specific technical method is shown in 3-1)
7-2 Conclusion
In order to further detect the sensitization of the compounds of the invention to the MEK inhibitor, trimetinib, in vitro, the invention selects the compounds of SY1-003、SY1-006、SY1-009、SY1-013、SY1-015、SY1-023、SY1-029、SY1-038、SY1-001、SY1-007、SY1-010、SY1-014、SY1-021、SY1-025、SY1-033、SY1-037、SY1-039 and the like with antiproliferative activity (IC 50) lower than 50nM in tumor cells with high oxidative phosphorylation level to further detect the effect of the combined trimetinib on the proliferation of lung cancer cells, and finds that the compounds all have the effect of increasing the sensitivity of the trimetinib in the tumor cells. Among these, the present invention mainly shows the experimental results of representative compounds SY 1-001. The experimental results are shown in fig. 4, and SY1-001 increases the cytotoxicity of the trimetinib in lung cancer cells H460 and H441 in a concentration gradient manner, which shows that SY1-001 has a remarkable sensitization effect on the trimetinib (fig. 4). Meanwhile, the invention discovers that SY1-001 can obviously increase the sensitivity of the trimetinib in the acquired trimetinib-resistant lung cancer cells A549/TR, and shows that SY-001 and other compounds have the potential of reversing the drug resistance of the trimetinib.
Example eight representative Compounds examples SY1-001 anti-tumor Effect of the in vivo combination with the MEK inhibitor trametinib
8-1, The technical method comprises the following steps:
(1) Subcutaneous tumor-bearing experiment of nude mice
H460 cells were digested and counted and resuspended to 3×10 6 cells per 100 μl with serum-free DMEM medium. After the cells are blown and evenly mixed, the cell suspension is sucked by a 1ml syringe and injected into the subcutaneous of 5-6 weeks nude mice, and 100 microliter of cell suspension is injected into each nude mouse, so as to establish a tumor cell subcutaneous tumor-bearing model of human sources. When the tumor grows to a certain volume (about 150 cubic millimeters), the tumor is divided into four groups (control group, trimetinib-1 mg/kg, SY-001-30 mg/kg and combined group) on average according to the size of the tumor, and medicines are given to the SY1-001, the trimetinib and the combined treatment of the SY1-001 and the trimetinib prepared by the embodiment of the invention. The compounds were formulated with 0.5% sodium methylcellulose (CMC-Na) and administered orally by gavage, and tumor volumes (volume = length x width 2 x 0.52) were measured every 3 days while the body weights of the mice were measured. After the tumor volume reached around 2000 cubic millimeters, mice were sacrificed. GraphPad8 Prism software was used for data statistics. (other technical methods of representing compound detection of this publication are similar to SY 1-001)
8-2 Conclusion
In the H460 xenogeneic model, the present invention evaluates a number of representative compounds, such as SY1-001, SY1-007, SY1-010, SY1-017, SY1-024, SY1-028, for in vivo antitumor activity and finds that such compounds have similar antitumor activity. Among them, the experimental results of SY1-001 were selected for discussion. As shown in fig. 5, SY1-001 significantly inhibited tumor growth, with significant advantages over the control group (fig. 5A, 5B, and 5C); meanwhile, SY1-001 (30 mg/kg, twice a day) and trametinib (5 mg/kg, once a day) are combined to inhibit tumors better than SY1-001 or trametinib single drug treatment, which shows that SY1-001 can significantly enhance the anti-tumor effect of the trametinib in vivo, has the potential of overcoming MEK inhibitor resistance (figures 5A, 5B and 5C), in addition, the invention carries out in vivo toxicity evaluation on SY1-001, and the study finds that the use of SY1-001 alone or in combination with the trametinib does not lead to significant weight loss of mice, and the compound has better safety (figure 5D).
EXAMPLE nine preparation of representative Compounds of the invention
Example 9 Synthesis of 1, N-ethyl-5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) pyridine amide (SY 1-002).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethylaniline into ethylamine, and the yield is high ：20％.1H NMR(500MHz,Chloroform-d)δ8.50–8.45(m,1H),8.23(dd,J＝8.1,4.2Hz,1H),7.94(s,1H),7.74–7.67(m,1H),7.40–7.35(m,1H),7.01–6.93(m,3H),5.60(d,J＝9.3Hz,2H),4.43–4.34(m,2H),3.88(d,J＝22.8Hz,1H),3.79(s,5H),3.58–3.49(m,4H),2.49(d,J＝14.8Hz,3H),1.36(s,2H),1.31(s,3H).
Example 9 Synthesis of 2, N-isobutyl-5- (5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazole-1-methyl) pyridine amide (SY 1-003).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethylaniline into isopropylamine, and the yield is high ：30％.1H NMR(500MHz,Chloroform-d)δ8.46(d,J＝2.3Hz,1H),8.22(d,J＝8.0Hz,1H),8.05(s,1H),7.70(dd,J＝8.1,2.3Hz,1H),7.32(d,J＝8.1Hz,2H),6.96–6.90(m,2H),5.59(s,2H),4.37(q,J＝8.1Hz,2H),4.15(s,2H),3.81(s,3H),3.56(s,2H),3.32(t,J＝6.6Hz,2H),2.58(s,4H),2.47(s,3H),1.00(d,J＝6.7Hz,6H).
Example 9-3,5 Synthesis of- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (tetrahydro-2H-pyran-4-yl) pyridine amide (SY 1-004).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethylaniline into 4-aminotetrahydropyran, and the yield is high ：27％.1H NMR(500MHz,Chloroform-d)δ8.48(dd,J＝7.9,2.2Hz,1H),8.22(dd,J＝8.0,5.5Hz,1H),7.91(d,J＝8.4Hz,1H),7.71(dd,J＝8.1,2.3Hz,1H),7.40–7.30(m,4H),5.60(d,J＝9.7Hz,2H),4.38(q,J＝8.1Hz,2H),4.03(dt,J＝11.5,3.5Hz,2H),3.82(d,J＝31.6Hz,2H),3.63–3.49(m,3H),2.59(s,3H),2.49(d,J＝16.2Hz,3H),2.01(d,J＝12.9Hz,2H),1.66(qd,J＝11.4,4.5Hz,3H),1.35(s,1H),1.31(s,1H),1.28(s,2H).
Example 9-4,5 Synthesis of- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (2- (trifluoromethyl) phenyl) pyridine amide (SY 1-005).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethylaniline into the ortho-trifluoromethylaniline, and the yield is high ：53％.1H NMR(500MHz,Chloroform-d)δ10.54(s,1H),8.63–8.55(m,2H),8.32(d,J＝8.0Hz,1H),7.78(dd,J＝8.1,2.2Hz,1H),7.69(d,J＝7.9Hz,1H),7.64(t,J＝7.9Hz,1H),7.31(d,J＝17.9Hz,3H),6.98–6.90(m,2H),5.63(s,2H),4.37(q,J＝8.1Hz,2H),4.24–4.08(m,2H),3.81(s,2H),3.57(s,2H),2.50(s,7H).
Example 9-5,5 Synthesis of- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (3- (trifluoromethyl) phenyl) pyridine amide (SY 1-006).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethylaniline into m-aminotrifluorotoluene, and the yield is high ：55％.1H NMR(500MHz,Chloroform-d)δ10.05(s,1H),8.56(d,J＝2.2Hz,1H),8.32(d,J＝8.0Hz,1H),8.10(d,J＝2.0Hz,1H),7.97(dt,J＝8.3,1.5Hz,1H),7.78(dd,J＝8.1,2.2Hz,1H),7.53(t,J＝8.0Hz,1H),7.44(d,J＝7.8Hz,1H),7.35(s,2H),6.95(d,J＝8.1Hz,2H),5.63(s,2H),4.37(q,J＝8.1Hz,2H),4.15(s,1H),3.82(s,2H),3.60(d,J＝14.5Hz,2H),2.59(s,5H),2.50(s,3H).
Example 9-6 Synthesis of N- (4-fluorophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) pyridine amide (SY 1-007).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethylaniline into 4-fluoroaniline, and the yield is high ：12％.1H NMR(500MHz,Chloroform-d)δ9.88(s,1H),8.55(dd,J＝8.4,2.2Hz,1H),8.36–8.30(m,1H),7.81–7.71(m,3H),7.67(s,1H),7.40–7.33(m,1H),7.15–7.07(m,2H),7.04(s,1H),6.95(d,J＝8.7Hz,1H),5.63(d,J＝14.3Hz,2H),4.40(q,J＝8.2Hz,2H),4.30(s,1H),4.14(d,J＝38.1Hz,2H),3.83(d,J＝33.5Hz,3H),3.57(d,J＝51.6Hz,2H),3.13–2.70(m,2H),2.53(s,1H),2.51(s,2H).
Example 9-7 Synthesis of N- (4-chlorophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide (SY 1-008).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethylaniline into 4-chloroaniline, and the yield is high ：18％.1H NMR(500MHz,DMSO-d6)δ10.81(s,1H),8.69(d,J＝2.2Hz,1H),8.16(d,J＝8.1Hz,1H),8.00–7.93(m,2H),7.85(dd,J＝8.1,2.3Hz,1H),7.46–7.40(m,2H),7.28(d,J＝8.3Hz,2H),7.02(d,J＝8.3Hz,2H),5.82(s,2H),4.74(q,J＝8.9Hz,2H),3.85(s,2H),3.62(s,2H),3.47(s,2H),2.52(s,2H),2.50(s,2H),2.39(s,3H).
Example 9-8 Synthesis of N- (4-bromophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide (SY 1-009).
The same synthesis method as (SY 1-001) is adopted to change the para-trifluoromethyl aniline into 4-bromoaniline, and the yield is high ：41％.1H NMR(500MHz,Chloroform-d)δ9.92(s,1H),8.55(dd,J＝8.3,2.2Hz,1H),8.32(dd,J＝8.1,4.0Hz,1H),7.77(dd,J＝8.1,2.2Hz,1H),7.72–7.66(m,3H),7.56–7.50(m,2H),7.02–6.92(m,3H),5.64(d,J＝9.1Hz,2H),4.38(q,J＝8.0,3.4Hz,2H),4.12(s,2H),3.83(d,J＝32.5Hz,4H),3.64–3.28(m,2H),3.02(s,2H),2.53(s,1H),2.50(s,2H).
Example 9-9, synthesis of 5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethoxy) phenyl) pyridinecarboxamide (SY 1-010).
The same synthesis method as (SY 1-001) is adopted to convert the para-trifluoromethylaniline into para-trifluoromethoxyaniline, and the yield is high ：60％.1H NMR(500MHz,Chloroform-d)δ10.01(s,1H),8.56(s,1H),8.35(d,J＝8.0Hz,1H),7.81(dd,J＝10.9,8.6Hz,3H),7.67(d,J＝8.4Hz,2H),7.26(s,2H),7.05(d,J＝8.2Hz,2H),5.68–5.58(m,2H),4.40(q,J＝7.9Hz,2H),4.24–4.04(m,2H),3.82–3.72(m,2H),3.52(s,2H),3.06(s,2H),2.81(s,2H),2.51(s,3H).
Examples 9-10,5 Synthesis of- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (naphthalen-2-yl) pyridine amide (SY 1-011).
The same synthesis as (SY 1-001) was used to convert p-trifluoromethylaniline to 2-naphthylamine to give 24mg of the product 5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (naphthalen-2-yl) pyridine amide (SY 1-011), yield ：91％.1H NMR(500MHz,Chloroform-d)δ10.09(s,1H),8.57(d,J＝2.2Hz,1H),8.50(d,J＝2.1Hz,1H),8.36(d,J＝8.0Hz,1H),7.88(dd,J＝8.5,3.8Hz,2H),7.83(d,J＝8.1Hz,1H),7.78(dd,J＝8.1,2.2Hz,1H),7.71(dd,J＝8.8,2.2Hz,1H),7.54–7.48(m,1H),7.48–7.42(m,1H),7.33(s,2H),6.98–6.90(m,2H),5.63(s,2H),4.37(q,J＝8.1Hz,2H),3.83(d,J＝27.9Hz,2H),3.56(s,2H),2.56(d,J＝16.9Hz,6H),2.51(s,3H).
Examples 9-11,5 Synthesis of- ((5-methyl-4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-012).
The same synthetic procedure as (SY 1-001) was used to replace 1- (4- (2, 2-trifluoroethoxy) benzyl) piperazine hydrochloride with 1- (2, 2-trifluoroethoxy) benzyl) piperazine hydrochloride to give the product 5- ((5-methyl-4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) picolinamide (SY 1-012) in 76% yield. Yield rate ：91％.1H NMR(500MHz,Chloroform-d)δ10.08(s,1H),8.60–8.53(m,1H),8.34(dd,J＝8.2,3.6Hz,1H),7.91(d,J＝8.4Hz,2H),7.79(dd,J＝8.1,2.3Hz,1H),7.67(d,J＝8.5Hz,2H),7.54–7.37(m,2H),7.21(s,1H),6.98–6.85(m,1H),5.65(d,J＝10.3Hz,2H),4.53–4.40(m,2H),4.32(s,2H),4.14(s,1H),3.92–3.72(m,2H),3.54(s,1H),3.22–2.77(m,4H),2.52(d,J＝14.2Hz,3H).
Examples 9-12, synthesis of 5- ((5-methyl-4- (4- (3- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-013)
The same synthesis as (SY 1-001) was used to convert 1- (4- (2, 2-trifluoroethoxy) benzyl) piperazine hydrochloride to 1- (3- (2, 2-trifluoroethoxy) benzyl) piperazine hydrochloride to give the target product 5- ((5-methyl-4- (4- (3- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-013), yield ：45％.1H NMR(500MHz,Chloroform-d)δ10.08(s,1H),8.56(d,J＝2.3Hz,1H),8.36–8.31(m,1H),7.91(d,J＝8.4Hz,2H),7.79(dd,J＝8.1,2.3Hz,1H),7.67(d,J＝8.5Hz,2H),7.28(s,2H),7.10(s,2H),5.64(s,2H),4.44(q,2H),4.14(s,2H),3.78(d,J＝19.2Hz,3H),3.51(d,J＝5.2Hz,1H),3.03(d,J＝12.1Hz,1H),2.64(s,3H),2.51(s,3H).
Examples 9-13,5 synthesis of- ((5-methyl-4- (4- (4- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-014).
1- (4- (2, 2-Trifluoroethoxy) benzyl) piperazine hydrochloride was converted to 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride by the same synthetic method as (SY 1-001), yield ：79％.1H NMR(500MHz,Chloroform-d)δ10.08(s,1H),8.56(d,J＝2.2Hz,1H),8.33(d,J＝8.1Hz,1H),7.91(d,J＝8.5Hz,2H),7.79(dd,J＝8.1,2.3Hz,1H),7.67(d,J＝8.5Hz,2H),7.41(s,2H),7.23(s,2H),5.64(s,2H),4.15(s,2H),3.80(s,2H),3.58(s,2H),2.61(d,J＝28.7Hz,4H),2.50(s,3H).
Examples 9-14,5 Synthesis of- ((5-methyl-4- (4- (4- (trifluoromethoxy) benzoyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-015).
1- (4- (2, 2-Trifluoroethoxy) benzyl) piperazine hydrochloride was converted to piperazin-1-yl (4- (trifluoromethoxy) phenyl) methanone hydrochloride using the same synthetic method as (SY 1-001), yield ：79％.1H NMR(500MHz,Chloroform-d)δ10.08(s,1H),8.60–8.55(m,1H),8.34(d,J＝8.1Hz,1H),7.91(d,J＝8.5Hz,2H),7.80(d,J＝7.9Hz,1H),7.67(d,J＝8.5Hz,2H),7.55–7.48(m,2H),7.31(d,J＝8.3Hz,2H),5.65(s,2H),3.74(d,J＝129.9Hz,8H),2.53(s,3H).
Examples 9-15,5 Synthesis of- ((4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-016).
The same synthesis method as (SY 1-001) is adopted to change the ethyl 2-butynoate into ethyl propiolate, and the yield is high ：90％.1H NMR(500MHz,Chloroform-d)δ10.15(s,1H),8.61(d,J＝2.2Hz,1H),8.35(d,J＝8.0Hz,1H),7.96(d,J＝8.4Hz,2H),7.90(d,J＝8.2Hz,2H),7.80(d,J＝7.9Hz,2H),7.71(d,J＝8.4Hz,2H),7.37(s,2H),5.80(s,2H),4.38(q,2H),3.71(s,2H),3.38(s,2H),3.14(s,2H),2.33(s,2H),1.77(s,2H).
Examples 9-16, synthesis of 5- ((5-ethyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-017)
The same synthesis method as (SY 1-001) is adopted to change the ethyl 2-butynoate into ethyl 2-pentynoate, and the yield is high ：81％.1H NMR(500MHz,Chloroform-d)δ10.08(s,1H),8.57(dd,J＝10.4,2.2Hz,1H),8.34(d,J＝8.1Hz,1H),7.91(d,J＝8.4Hz,2H),7.84–7.78(m,1H),7.67(d,J＝8.5Hz,3H),7.38(d,J＝8.6Hz,1H),7.04(s,1H),6.98–6.93(m,1H),5.66(s,2H),4.38(q,2H),3.83(d,J＝33.3Hz,3H),3.51(s,1H),1.59(s,11H).
Examples 9-17,5 Synthesis of- ((4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -5- (trifluoromethyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-018)
The same synthesis method as (SY 1-001) is adopted to change the ethyl 2-butynoate into the ethyl 4, 4-trifluoro-2-butynoate, and the yield is high ：35％.1H NMR(500MHz,CDCl3)δ10.08(s,1H),8.56(d,J＝1.7Hz,1H),8.32(d,J＝8.1Hz,1H),7.91(d,J＝8.5Hz,2H),7.79(dd,J＝8.1,2.2Hz,1H),7.66(d,J＝8.6Hz,2H),5.64(s,2H),4.23(s,2H),3.88(s,2H),2.66(d,J＝12.8Hz,5H),2.61–2.45(m,7H).
Examples 9-18,5 Synthesis of- ((5-phenyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-019)
The same synthesis method as (SY 1-001) is adopted to change the ethyl 2-butynoate into ethyl phenylpropionate, and the yield is high ：12％.1H NMR(500MHz,Chloroform-d)δ10.05(s,1H),8.33(d,J＝2.1Hz,1H),8.23(d,J＝8.0Hz,1H),7.90(d,J＝8.4Hz,2H),7.65(dd,J＝12.8,8.3Hz,3H),7.58–7.47(m,3H),7.32–7.28(m,3H),7.27(s,1H),6.95–6.89(m,2H),5.58(s,2H),4.36(q,J＝8.1Hz,2H),3.78(d,J＝52.5Hz,4H),3.56–3.42(m,2H),2.47(d,J＝11.4Hz,4H).
Examples 9-19, synthesis of 5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethoxy) phenyl) pyridinecarboxamide (SY 1-020)
5- (Methoxycarbonyl) pyridine-2-carboxylic acid (1.0 g,5.52 mmol), 4- (trifluoromethoxy) aniline (1.2 g,6.62 mmol), HATU (2.5 g,6.62 mmol) were weighed in a round bottom flask, then 10mL DMF was added, DIEA (2.9 mL,16.56 mmol) was slowly added dropwise, reacted for 2h at normal temperature, water was added and suction filtered, the resulting solid was dried over anhydrous Na 2SO4, then column chromatography purification gave intermediate 6- ((4- (trifluoromethoxy) phenyl) carbamoyl) methylnicotinate 901mg, yield 48%; intermediate 6- ((4- (trifluoromethoxy) phenyl) carbamoyl) methylnicotinate (901 mg,2.64 mmol) was then dissolved in 20mL THF solution, nitrogen was placed in a low temperature reactor, stirred to dissolve and drop LiAlH 4 (5 mL,5.29 mmol) at-20 ℃ reacted for 1-2h at-20 ℃, ice water was added dropwise to quench LiAlH 4, the funnel-pad celite was suction filtered, ethyl acetate rinsed LiAlH 4 colloid, the filtrate was spin-dried by spin-evaporation, and column chromatography was purified to give intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethoxy) phenyl) picolinamide 347mg, yield 42%.
Intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethoxy) phenyl) pyridine amide (347mg, 1.11 mmol) was dissolved in 20mL DCM, stirred in ice bath, PBr 3 (0.08 mL,0.83 mmol) was added dropwise under ice bath, reacted for 1-2h under ice bath, PBr 3 was quenched by ice dropwise, extracted, dried and stirred through column to give intermediate 5- (bromomethyl) -N- (4- (trifluoromethoxy) phenyl) pyridine amide 416mg in 100% yield; intermediate 5- (bromomethyl) -N- (4- (trifluoromethoxy) phenyl) picolinamide (416 mg,1.10 mmol), sodium azide (87 mg,1.33 mmol) was weighed out in 4mL DMF, reacted for 2h, extracted with EA, stripped with saturated NaCl and dried to give intermediate 5- (azidomethyl) -N- (4- (trifluoromethoxy) phenyl) picolinamide 157mg in 42% yield.
Intermediate 5- (azidomethyl) -N- (4- (trifluoromethoxy) phenyl) pyridine amide (157 mg,0.46 mmol) and ethyl butynoate (313 mg,2.79 mmol) were weighed, 12mL of toluene solution was added to react overnight at 125 ℃, toluene was removed by rotary evaporation, EA extraction, drying, column chromatography purification gave intermediate 4-methyl-1- ((6- ((4- (trifluoromethoxy) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid ethyl ester 105mg, yield 50%; then intermediate 4-methyl-1- ((6- ((4- (trifluoromethoxy) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid ethyl ester (105 mg,0.23 mmol) was dissolved in 15ml CH 3OH:H2 O=4:1 mixed solvent, liOH.H 2 O (39 mg,0.93 mmol) was added, stirring 1H at room temperature, spinning dry methanol solvent at 45 ℃, dropping concentrated hydrochloric acid to adjust pH to acidity, EA extraction, drying to give intermediate 4-methyl-1- ((6- ((4- (trifluoromethoxy) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid 98mg, 100% yield.
The intermediate 4-methyl-1- ((6- ((4- (trifluoromethoxy) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid (98 mg,0.23 mmol), 1- (4- (2, 2-trifluoroethoxy) benzyl) piperazine hydrochloride (86 mg,0.27 mmol), HATU (106 mg,0.27 mmol) were dissolved in DMF solution, DIEA (0.12 mL,0.69 mmol) was added, reacted overnight at normal temperature, EA extracted, saturated NaCl back extracted three times, dried, column chromatographed to give the title product 5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazole-1-yl) methyl) -N- (4- (trifluoromethoxy) phenyl) pyridine carboxamide SY1-020 (60 mg), yield ：39％.1H NMR(500MHz,Chloroform-d)δ10.13(s,1H),8.65(s,1H),8.42(d,J＝7.9Hz,1H),8.01(d,J＝7.9Hz,1H),7.92–7.86(m,2H),7.56(d,J＝8.1Hz,2H),7.31(s,2H),6.84(d,J＝8.1Hz,2H),5.93(s,2H),4.30(q,J＝8.0Hz,2H),3.90(s,4H),3.39(s,4H),3.09(s,2H),2.33(s,3H).
Examples 9-20, synthesis of 5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N-phenylpyridine-amide (SY 1-021)
The same method as (SY 1-020) is adopted to change the para-trifluoromethoxy aniline into aniline, and the yield is high ：77％.1H NMR(500MHz,DMSO-d6)δ10.72(s,1H),8.62(d,J＝2.1Hz,1H),8.20(d,J＝8.1Hz,1H),7.98–7.92(m,2H),7.84(dd,J＝8.1,2.2Hz,1H),7.42–7.35(m,2H),7.17–7.10(m,1H),7.10–7.03(m,2H),6.91–6.84(m,2H),5.77(d,J＝5.8Hz,2H),4.68(q,J＝8.9Hz,2H),3.52(s,2H),3.18(s,2H),3.06(s,2H),2.53–2.51(m,2H),2.50(s,2H),2.21(s,3H).
Examples 9-21 Synthesis of N- (4-methoxyphenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide (SY 1-022)
The same method as (SY 1-020) is adopted to change the para-trifluoromethoxy aniline into 4-methoxy aniline, and the yield is high ：81％.1H NMR(500MHz,Chloroform-d)δ9.94(s,1H),8.61(s,1H),8.41(d,J＝8.0Hz,1H),7.97(d,J＝7.9Hz,1H),7.78–7.72(m,2H),7.54(d,J＝8.1Hz,2H),6.97(d,J＝9.0Hz,2H),6.77(d,J＝8.1Hz,2H),5.80(dd,J＝40.7,19.0Hz,2H),4.26(q,J＝8.0Hz,2H),3.91(s,2H),3.84(s,3H),3.35(s,3H),2.99(s,1H),2.31(s,3H),1.80(s,4H).
Examples 9-22 Synthesis of N- (4- (tert-butyl) phenyl) -5- (4-methyl-5- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazole-1-methyl) pyridine amide (SY 1-023)
The same method as (SY 1-020) is adopted to change the p-trifluoromethoxy aniline into 4- (tertiary butyl) aniline, and the yield is high ：70％.1H NMR(500MHz,DMSO-d6)δ10.67–10.63(m,1H),8.60(d,J＝2.0Hz,1H),8.20(dd,J＝8.1,1.6Hz,1H),7.84(td,J＝8.3,3.8Hz,3H),7.39(dd,J＝8.6,1.7Hz,2H),7.08–7.02(m,2H),6.88–6.82(m,2H),5.94–5.64(m,2H),4.67(q,J＝8.2Hz,2H),3.33(s,2H),3.17(s,2H),3.05(s,2H),2.51(s,2H),2.50(s,2H),2.20(d,J＝1.6Hz,3H),1.28(d,J＝1.6Hz,9H).
Examples 9-23, synthesis of 5- (4-methyl-5- (4- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (p-tolyl) pyridine amide (SY 1-024)
The same method as (SY 1-020) is adopted to convert the para-trifluoromethoxy aniline into the para-toluidine, and the yield is high :18％.1H NMR(500MHz,DMSO-d6)δ10.64(s,1H),8.61(d,J＝2.1Hz,1H),8.19(d,J＝8.1Hz,1H),7.83(dd,J＝8.5,2.5Hz,3H),7.18(d,J＝8.1Hz,2H),7.07(d,J＝8.3Hz,2H),6.87(d,J＝8.4Hz,2H),5.77(s,2H),4.68(q,J＝8.9Hz,2H),3.33–3.30(m,2H),3.17(s,2H),3.05(s,2H),2.50(s,4H),2.29(s,3H),2.20(s,3H).
Examples 9-24 Synthesis of N- (4-methoxy-3- (trifluoromethyl) phenyl) -5- (4-methyl-5- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methylpyridine amide (SY 1-025)
The same procedure as in (SY 1-020) was used to convert p-trifluoromethoxy aniline to 4-methoxy-3- (trifluoromethyl) aniline in the yield ：18％.1H NMR(500MHz,DMSO-d6)δ10.97(s,1H),8.63(d,J＝2.2Hz,1H),8.34(d,J＝2.7Hz,1H),8.20(dd,J＝8.4,3.8Hz,2H),7.83(dd,J＝8.1,2.2Hz,1H),7.30(d,J＝9.1Hz,1H),7.10–7.05(m,2H),6.91–6.85(m,2H),5.77(d,J＝5.6Hz,2H),4.68(q,J＝8.9Hz,2H),3.88(s,3H),3.30(s,2H),3.18(s,2H),3.06(s,2H),2.52(s,4H),2.21(s,3H).
Examples 9-25 Synthesis of N- (4-fluorophenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) pyridine amide (SY 1-026)
The same method as (SY 1-020) is adopted to change the para-trifluoromethoxy aniline into 4-fluoroaniline, and the yield is high ：58％.1H NMR(500MHz,Chloroform-d)δ9.98(s,1H),8.62(s,1H),8.39(d,J＝7.8Hz,1H),7.96(d,J＝7.9Hz,1H),7.80(dd,J＝9.0,4.6Hz,2H),7.55(d,J＝7.9Hz,2H),7.13(t,J＝8.5Hz,2H),6.83(d,J＝7.8Hz,2H),5.85(d,J＝74.5Hz,2H),4.30(q,J＝8.0Hz,2H),3.86(s,4H),3.38(s,4H),3.06(s,2H),2.32(s,3H).
Examples 9-26 Synthesis of N- (4-bromophenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide (SY 1-027)
The same method as (SY 1-020) is adopted to change the para-trifluoromethoxy aniline into 4-bromoaniline, and the yield is high ：32％.1H NMR(500MHz,DMSO-d6)δ10.91(s,1H),8.63(d,J＝2.1Hz,1H),8.20(d,J＝8.1Hz,1H),7.99–7.92(m,2H),7.83(dd,J＝8.1,2.2Hz,1H),7.60–7.53(m,2H),7.09–7.04(m,2H),6.92–6.87(m,2H),5.76(s,2H),4.69(q,J＝8.8Hz,2H),3.52(s,3H),3.18(s,2H),3.06(s,2H),2.51–2.48(m,3H),2.21(s,3H).
Examples 9-27,5 Synthesis of- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridinecarboxamide (SY 1-028)
The same procedure as in (SY 1-020) was used to convert 4-bromoaniline into para-trifluoromethylaniline in the yield ：39％.1H NMR(500MHz,DMSO-d6)δ11.15(s,1H),8.65(d,J＝2.2Hz,1H),8.22(dd,J＝8.4,6.2Hz,3H),7.85(dd,J＝8.1,2.2Hz,1H),7.76(d,J＝8.6Hz,2H),7.07–7.02(m,2H),6.90–6.83(m,2H),5.78(d,J＝9.0Hz,2H),4.68(q,J＝8.9Hz,2H),3.51(s,2H),3.17(s,2H),3.06(s,2H),2.51(s,2H),2.50(s,2H),2.21(s,3H).
Examples 9-28, synthesis of 5- ((4-methyl-5- (4- (4- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-029)
5- (Methoxycarbonyl) -2-pyridinecarboxylic acid (5.0 g,27.60 mmol), para-trifluoromethylaniline (5.3 g,33.12 mmol), HATU (12.5 g,33.12 mmol) were weighed in a 250mL round bottom flask and dissolved in 20mL DMF, DIEA (14 mL,82.80 mmol) was slowly added dropwise, reacted at normal temperature for 2h, filtered with water and suction, the resulting solid was dried over anhydrous Na 2SO4, and then the solid was slurried with a small amount of ethyl acetate to give intermediate 6- ((4- (trifluoromethyl) phenyl) carbamoyl) nicotinic acid methyl ester 4.0g, 78% yield; then, intermediate 6- ((4- (trifluoromethyl) phenyl) carbamoyl) methylnicotinate (4.0 g,13.73 mmol) was dissolved in 20mL THF solution, air was replaced with nitrogen, and the mixture was placed in a low temperature reactor and stirred at-20℃for three to five minutes, 2eq LiAlH 4 (27 mL,27.46 mmol) was added dropwise to the reaction system, reacted at-20℃for 1-2 hours, ice water was added dropwise to quench LiAlH 4, the funnel pad of the sand was suction-filtered, ethyl acetate was used to rinse LiAlH 4 colloid, the filtrate was dried by rotary evaporation, and purified by column chromatography to give intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 2.4g in 59% yield.
Intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (2.4 g,8.25 mmol) was dissolved in 20mL DCM, stirred in ice bath, PBr 3 (0.6 mL,6.19 mmol) was added dropwise under ice bath, reacted for 1-2h under ice bath, PBr 3 was quenched dropwise with ice water, extracted, dried, and purified by column chromatography to give intermediate 5- (bromomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 593mg, 20% yield; intermediate 5- (bromomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (593 mg,1.65 mmol) was weighed out, naN 3 (129 mg,1.98 mmol) was dissolved in 4mL DMF solution, stirred at normal temperature for 2h, EA extracted, saturated NaCl back extracted, dried to give intermediate 5- (azidomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 476mg, 90% yield.
Intermediate 5- (azidomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (470 mg,1.48 mmol) and ethyl 2-butynoate (1.0 g,8.92 mmol) were dissolved in 12mL toluene, heated to 120 ℃, reacted overnight, dried toluene, extracted with water, dried, purified by column chromatography to give intermediate 4-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylate 131mg, 21% yield; intermediate 5-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-4-carboxylic acid methyl ester (131 mg,0.31 mmol) was then dissolved in 20ml ch 3OH:H2 o=4:1 mixed solvent, lioh.h 2 O (52 mg,1.24 mmol) was added, reacted for 1H at room temperature, dried methanol was brought to acidity with concentrated hydrochloric acid solution, EA extracted and the dried solid afforded intermediate 4-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid 114mg, 90% yield.
Intermediate 4-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid (114 mg,0.28 mmol), 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride (100 mg,0.33 mmol), HATU (127 mg,0.33 mmol) were weighed out in DMF solution, DIEA (0.14 ml,0.84 mmol) was added, reacted overnight, post-treated and purified by column chromatography to give the target product 5- ((4-methyl-5- (4- (4- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-029) 36mg, yield ：20％.1H NMR(600MHz,DMSO-d6)δ11.12(s,1H),8.64(d,J＝2.2Hz,1H),8.22(t,J＝8.6Hz,3H),7.87(dd,J＝8.1,2.2Hz,1H),7.76(d,J＝8.5Hz,2H),7.20(d,J＝8.3Hz,2H),7.15(d,J＝8.2Hz,2H),5.79(s,2H),3.30(s,2H),3.26(s,2H),3.07(s,2H),2.51(s,2H),2.50(s,2H),2.21(s,3H).
Examples 9-29, synthesis of 5- ((4-methyl-5- (4- (4-methylbenzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-030)
The same procedure as in (SY 1-029) was used to convert 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride to 1- (4-methylbenzyl) piperazine hydrochloride in the same yield ：25％.1H NMR(600MHz,DMSO-d6)δ11.11(s,1H),8.64(d,J＝2.2Hz,1H),8.22(dd,J＝8.3,6.0Hz,3H),7.85(dd,J＝8.1,2.2Hz,1H),7.76(d,J＝8.5Hz,2H),6.97(d,J＝1.9Hz,4H),5.78(s,2H),3.50(s,3H),3.17(s,2H),3.06(s,2H),2.51(s,3H),2.21(d,J＝2.3Hz,6H).
Examples 9-30, synthesis of 5- ((5- (4- (4-chloro-3- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-031)
The same procedure as in (SY 1-029) was used to convert 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride to 1- (4-chloro-3- (trifluoromethoxy) benzyl) piperazine hydrochloride in the yield ：25％.1H NMR(500MHz,Chloroform-d)δ10.10(s,1H),8.70–8.59(m,1H),8.37(d,J＝8.0Hz,1H),7.96(d,J＝8.4Hz,4H),7.70(d,J＝8.5Hz,3H),7.21(s,1H),5.82(s,2H),4.25–3.55(m,5H),3.14(d,J＝163.1Hz,5H),2.33(s,3H).
Examples 9-31, synthesis of 5- ((5- (4- (4-isopropylbenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-032)
The same procedure as in (SY 1-029) was used to convert 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride to 1- (4-isopropylbenzyl) piperazine hydrochloride in the yield ：12％.1H NMR(500MHz,Chloroform-d)δ10.22(s,1H),8.65(s,1H),8.42(d,J＝8.0Hz,1H),7.99(t,J＝8.3Hz,3H),7.69(d,J＝8.5Hz,2H),7.45(d,J＝7.7Hz,2H),7.13(d,J＝7.6Hz,2H),5.83(s,2H),3.92(s,4H),2.88–2.84(m,1H),2.33(s,3H),1.28(s,6H),1.20(s,3H),1.18(s,3H).
Examples 9-32,5- ((5- (4- (4- (tert-butyl) benzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-033) Synthesis
The same procedure as in (SY 1-029) was used to convert 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride to 1- (4- (tert-butyl) benzyl) piperazine hydrochloride in the yield ：53％.1H NMR(500MHz,DMSO-d6)δ11.11(s,1H),8.63–8.59(m,1H),8.27–8.21(m,3H),7.88(dd,J＝8.1,2.2Hz,1H),7.77(d,J＝8.6Hz,2H),7.15–7.10(m,2H),6.99–6.94(m,2H),5.79(s,2H),3.16(s,2H),3.05(s,2H),2.51(d,J＝1.9Hz,4H),2.50(s,2H),2.21(s,3H),1.19(s,9H).
Examples 9-33,5- ((5- (4- (4-chlorobenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-034) Synthesis
The same procedure as in (SY 1-029) was used to convert 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride to 1- (4-chlorobenzyl) piperazine hydrochloride in the same yield ：94％.1H NMR(500MHz,Chloroform-d)δ10.13(s,1H),8.60(d,J＝2.2Hz,1H),8.34(d,J＝8.0Hz,1H),7.94(d,J＝8.4Hz,2H),7.90(d,J＝8.3Hz,1H),7.69(d,J＝8.5Hz,2H),7.14(d,J＝49.7Hz,4H),5.78(s,2H),3.70(s,2H),3.20(d,J＝78.3Hz,4H),2.36(d,J＝15.5Hz,2H),2.32(s,3H),1.67(s,2H).
Example 9 Synthesis of 5- (5- (4- (3-bromo-4-fluorobenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl-N- (4-trifluoromethyl) phenyl) pyridine amide (SY 1-035)
The same procedure as in (SY 1-029) was used to convert 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride to 1- (3-bromo-4-fluorobenzyl) piperazine hydrochloride in the yield ：36％.1H NMR(500MHz,Chloroform-d)δ10.13(s,1H),8.66(s,1H),8.39(d,J＝7.9Hz,1H),8.01–7.90(m,3H),7.70(d,J＝8.3Hz,2H),7.54–7.46(m,2H),7.38(d,J＝8.0Hz,1H),5.85(d,J＝19.3Hz,2H),3.89(s,2H),3.40(s,3H),3.17(s,3H),2.52(d,J＝51.4Hz,2H),2.34(s,3H).
Example 9-35,5 Synthesis of- ((5- (4-benzylpiperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazolyl) methyl) -N- (4-trifluoromethyl) phenylpyridine-amide (SY 1-036)
The same procedure as in (SY 1-029) was used to convert 1- (4- (trifluoromethoxy) benzyl) piperazine hydrochloride to 1-benzylpiperazine hydrochloride in the yield ：50％.1H NMR(500MHz,DMSO-d6)δ11.12(s,1H),8.65(d,J＝2.1Hz,1H),8.24–8.20(m,3H),7.85(dd,J＝8.1,2.2Hz,1H),7.76(d,J＝8.6Hz,2H),7.20–7.16(m,3H),7.09(dd,J＝6.7,2.9Hz,2H),5.79(s,2H),3.23(s,2H),3.08(s,2H),2.51(s,3H),2.50(s,3H),2.21(s,3H).
Examples 9-36,5- ((4-methyl-5- (4- (methylsulfonyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-037) Synthesis
The intermediate 4-methyl-1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid (13 mg,0.03 mmol), 1- (methylsulfonyl) piperazine (6 mg,0.04 mmol), HATU (15 mg,0.04 mmol) were weighed out in 4mL DMF, N-diisopropylethylamine (0.02 mL,0.09 mmol) was added and stirred at ambient temperature for 2H to give the target product 5- ((4-methyl-5- (4- (methylsulfonyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-037) 6mg, purified by column chromatography in the same manner as (SY 1-029) to give the target product 6mg, yield 38％.1H NMR(500MHz,Chloroform-d)δ10.04(s,1H),8.62(s,1H),8.32(d,J＝8.0Hz,1H),7.92–7.88(m,3H),7.68(d,J＝8.4Hz,2H),5.80(s,2H),3.81(s,4H),3.22(d,J＝75.8Hz,4H),2.67(s,3H),2.36(s,3H).
Examples 9-37, synthesis of 5- ((5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -4- (trifluoromethyl) -1H-1,2, 3-triazol-1-yl) methyl) -N-4- (trifluoromethyl) phenyl) pyridine amide (SY 1-038)
5- (Methoxycarbonyl) -2-pyridinecarboxylic acid (102 mg,0.56 mmol), para-trifluoromethylaniline (108 mg,0.67 mmol), HATU (256 mg,0.67 mmol) were weighed into a 100mL round bottom flask, N-diisopropylethylamine (0.30 mL,1.68 mmol) was added, stirred at room temperature for 2h, appropriate amount of water was added, EA was extracted, saturated NaCl back extracted, dried over anhydrous Na 2SO4, and purified by column chromatography to give intermediate, methyl 6- ((4- (trifluoromethyl) phenyl) carbamoyl) nicotinic acid 142mg, yield 78%; then, intermediate 6- ((4- (trifluoromethyl) phenyl) carbamoyl) methylnicotinate (142 mg,0.44 mmol) was dissolved in 20mL THF solution, air was replaced with nitrogen, and the mixture was placed in a low temperature reactor and stirred at-20 ℃ for three to five minutes, 2eq LiAlH 4 (1.0 mL,0.88 mmol) was added dropwise to the reaction system, reacted at-20 ℃ for 1-2h, ice water was added dropwise to quench LiAlH 4, the funnel pad of the sand was suction-filtered with celite, ethyl acetate washed out of LiAlH 4 colloid, the filtrate was dried by rotary evaporation, and purified by column chromatography to give intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 77mg, 59% yield.
Intermediate 5- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (77 mg,0.25 mmol) was dissolved in 20mL DCM, stirred in ice bath, PBr 3 (0.02 mL,0.19 mmol) was added dropwise under ice bath, reacted for 1-2h under ice bath, PBr 3 was quenched dropwise with ice water, extracted, dried and purified by column chromatography to give intermediate 5- (bromomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 31mg, 20% yield; intermediate 5- (bromomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (31 mg,0.08 mmol) was weighed out, naN 3 (6.7 mg,0.10 mmol) was dissolved in 4mL DMF and reacted at room temperature for 2h, water was added, EA extraction, drying afforded intermediate 5- (azidomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide 27mg, 100% yield.
Intermediate 5- (azidomethyl) -N- (4- (trifluoromethyl) phenyl) pyridine carboxamide (27 mg,0.08 mmol) and ethyl 4, 4-trifluoro-2-butynate (15 mg,0.09 mmol) were dissolved in 10mL toluene solution, heated to 120 ℃, reacted overnight, dried toluene, extracted with water, and purified by column chromatography to give intermediate 4- (trifluoromethyl) -1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylate 20mg, 50% yield; intermediate 4- (trifluoromethyl) -1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid ethyl ester (20 mg,0.03 mmol) was then dissolved in 15ml ch 3OH:H2 o=4:1 mixed solvent, lioh. 2 O (7 mg,0.15 mmol) was added, stirred at room temperature for 1H, dried over methanol, PH adjusted to 1-2 by addition of concentrated hydrochloric acid solution, ea extraction, and dried to give intermediate 4- (trifluoromethyl) -1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid 16mg, 84% yield.
Intermediate 4- (trifluoromethyl) -1- ((6- ((4- (trifluoromethyl) phenyl) carbamoyl) pyridin-3-yl) methyl) -1H-1,2, 3-triazole-5-carboxylic acid (16 mg,0.03 mmol), 1- (4- (2, 2-trifluoroethoxy) benzyl) piperazine hydrochloride (12 mg,0.04 mmol), HATU (15 mg,0.04 mmol) were weighed out in DMF solution, DIEA (0.02 mL,0.09 mmol) was added dropwise, and after conventional work-up and column chromatography purification the overnight reaction gave the title product 5- ((5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -4- (trifluoromethyl) -1H-1,2, 3-triazole-1-yl) methyl) -N-4- (trifluoromethyl) phenyl) pyridine amide (SY 1-038) 16mg, yield ：76％.1H NMR(500MHz,Chloroform-d)δ10.13(s,1H),8.63(d,J＝2.2Hz,1H),8.39(d,J＝8.0Hz,1H),7.99–7.92(m,3H),7.70(d,J＝8.5Hz,2H),7.09(d,J＝7.5Hz,2H),6.77(d,J＝8.1Hz,2H),5.79(d,J＝116.3Hz,2H),4.29(q,J＝8.1Hz,2H),3.65–3.06(m,6H),2.78–2.44(m,2H),2.30–2.10(m,2H).
Examples 9-38,5 Synthesis of- ((4-ethyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-039)
The same procedure as in (SY 1-038) was followed except that ethyl 4, 4-trifluoro-2-butynoate was replaced with ethyl 2-valerate, yield ：35％.1H NMR(500MHz,Chloroform-d)δ10.13(s,1H),8.63(s,1H),8.39(d,J＝8.0Hz,1H),7.97(d,J＝8.4Hz,3H),7.70(d,J＝8.5Hz,2H),7.53(d,J＝8.2Hz,2H),6.81(d,J＝7.8Hz,2H),5.82–5.66(m,2H),4.28(q,J＝8.0Hz,2H),3.84(s,4H),3.22(d,J＝151.5Hz,4H),2.61(s,2H),1.38–1.30(m,3H),1.28(d,J＝2.4Hz,2H).
Examples 9-39,5 Synthesis of- ((4-phenyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide (SY 1-040)
The same procedure as in (SY 1-038) was followed except that ethyl 4, 4-trifluoro-2-butynate was converted to ethyl phenylpropionate in the yield ：18％.1H NMR(500MHz,Chloroform-d)δ10.15(s,1H),8.66(s,1H),8.36(d,J＝8.1Hz,1H),7.94(dd,J＝22.0,8.5Hz,3H),7.73–7.55(m,5H),7.46(d,J＝28.2Hz,4H),6.99(s,1H),6.72(s,1H),5.83(s,2H),4.36(q,2H),3.72(s,2H),3.13(s,3H),2.78(s,3H),2.65(s,2H).

Claims (15)

1. A triazole small molecule organic compound for specifically targeting mitochondria complex I to inhibit mitochondrial oxidative phosphorylation, which is characterized by comprising a compound shown as a formula (i) or related analogues or pharmaceutically acceptable salts, metabolites or prodrugs of the compound shown as the formula (i);
Wherein:
R 1 is selected from any one of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoroethoxy, trifluoromethyl, trifluoromethylethoxy, trifluoromethylpropoxy, trifluoromethylamino, C 1～C5 alkyl, C 1～C5 alkoxy;
r 2 is selected from any one of the following groups: hydrogen, C 1～C4 alkyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, phenyl, halophenyl, benzyl, phenethyl;
r 3 is selected from any one of the following groups: c 1～C6 alkyl, trifluoromethylphenyl, tetrahydropyran, halophenyl, trifluoromethoxyphenyl, C 5～C10 aryl, C 1～C3 alkoxyphenyl, C 2～C6 alkenyl, C 2～C6 alkynyl, C 1～C6 alkoxy, C 1～C6 alkylthio, C 3～C6 cycloalkyl, C 2～C6 cycloalkoxy, C 2～C6 cycloalkylthio, C 2～C6 nitrogen-containing cycloalkyl, 5-10 membered heteroaryl.
2. The small triazole class organic compound of claim 2, which specifically targets mitochondrial oxidative phosphorylation inhibition by mitochondrial complex i, and its related analogues or pharmaceutically acceptable salts, metabolites or prodrugs, wherein whenIn the 4-position of 1,2, 3-triazole, the structure is shown as (ii):
Wherein:
R 1 is selected from any one of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoroethoxy, trifluoromethyl, trifluoromethylethoxy, trifluoromethylpropoxy, trifluoromethylamino, C 1～C5 alkyl, C 1～C5 alkoxy;
R 2 is selected from any one of the following groups: hydrogen, C 1～C4 alkyl, trifluoromethyl, trifluoroethyl, phenyl, halophenyl, benzyl, phenethyl;
r 3 is selected from any one of the following groups: c 1～C6 alkyl, trifluoromethylphenyl, tetrahydropyran, halophenyl, trifluoromethoxyphenyl, C 5～C10 aryl, C 1～C3 alkoxyphenyl, C 2～C6 alkenyl, C 2～C6 alkynyl, C 1～C6 alkoxy, C 1～C6 alkylthio, C 3～C6 cycloalkyl, C 2～C6 cycloalkoxy, C 2～C6 cycloalkylthio, C 2～C6 nitrogen-containing cycloalkyl, 5-10 membered heteroaryl.
3. The triazole small molecule organic compound which specifically targets mitochondrial complex i to inhibit mitochondrial oxidative phosphorylation of claim 2, and its related analogues or pharmaceutically acceptable salts, metabolites or prodrugs, wherein when R 3 is an aromatic group, its structure is as shown in (iii):
Wherein:
R 1 is selected from any one of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoroethoxy, trifluoromethyl, trifluoromethylethoxy, trifluoromethylpropoxy, trifluoromethylamino, C 1～C5 alkyl, C 1～C5 alkoxy;
R 2 is selected from any one of the following groups: hydrogen, C 1～C4 alkyl, trifluoromethyl, trifluoroethyl, phenyl, halophenyl, benzyl, phenethyl;
r 4 is selected from any one of the following groups: hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethoxy, trifluoromethyl, trifluoromethylamino, C 1～C3 alkyl, C 1～C3 alkoxy;
Selected from 5-to 8-membered aromatic rings or aromatic heterocycles, taken from any one of the following aromatic rings: benzene ring, pyridine, pyridazine, pyrimidine, pyrazine, thiophene, thiazole, isothiazole, pyrrole, pyrazole, imidazole, oxazole, oxadiazole, 1,2, 3-triazole, 1,2, 4-triazole, naphthalene ring, phenanthrene ring, quinoline, isoquinoline, indole, benzodiazole, benzotriazol, purine.
4. A triazole class small molecule organic compound which inhibits mitochondrial oxidative phosphorylation, and related analogues or pharmaceutically acceptable salts, metabolites or prodrugs thereof, according to any of claims 1-3, wherein said compound is selected from the group consisting of:
5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
n-ethyl-5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) pyridine amide;
N-isobutyl-5- (5-methyl-4- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazole-1-methyl) pyridine amide;
5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (tetrahydro-2H-pyran-4-yl) picolinamide;
5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (2- (trifluoromethyl) phenyl) pyridine amide;
5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (3- (trifluoromethyl) phenyl) pyridine amide;
n- (4-fluorophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide;
N- (4-chlorophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide;
n- (4-bromophenyl) -5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide;
5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethoxy) phenyl) pyridinecarboxamide;
5- ((5-methyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (naphthalen-2-yl) picolinamide;
5- ((5-methyl-4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5-methyl-4- (4- (3- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5-methyl-4- (4- (4- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5-methyl-4- (4- (4- (trifluoromethoxy) benzoyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) picolinamide;
5- ((5-ethyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -5- (trifluoromethyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5-phenyl-4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethoxy) phenyl) pyridinecarboxamide;
5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N-phenylpyridine amide;
N- (4-methoxyphenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide;
N- (4- (tert-butyl) phenyl) -5- (4-methyl-5- (4- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazole-1-methyl) pyridine amide;
5- (4-methyl-5- (4- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (p-tolyl) pyridine amide;
N- (4-methoxy-3- (trifluoromethyl) phenyl) -5- (4-methyl-5- (4- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) picolinamide;
n- (4-fluorophenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide;
n- (4-bromophenyl) -5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) picolinamide;
5- ((4-methyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridinecarboxamide;
5- ((4-methyl-5- (4- (4- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((4-methyl-5- (4- (4-methylbenzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5- (4- (4-chloro-3- (trifluoromethoxy) benzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5- (4- (4-isopropylbenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5- (4- (4- (tert-butyl) benzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5- (4- (4-chlorobenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- (5- (4- (3-bromo-4-fluorobenzyl) piperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazol-1-yl) methyl-N- (4-trifluoromethyl) phenyl) picolinamide;
5- ((5- (4-benzylpiperazine-1-carbonyl) -4-methyl-1H-1, 2, 3-triazolyl) methyl) -N- (4-trifluoromethyl) phenylpyridine amide;
5- ((4-methyl-5- (4- (methylsulfonyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -4- (trifluoromethyl) -1H-1,2, 3-triazol-1-yl) methyl) -N-4- (trifluoromethyl) phenyl) pyridine amide;
5- ((4-ethyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide;
5- ((4-phenyl-5- (4- (4- (2, 2-trifluoroethoxy) benzyl) piperazine-1-carbonyl) -1H-1,2, 3-triazol-1-yl) methyl) -N- (4- (trifluoromethyl) phenyl) pyridine amide.
5. A pharmaceutical composition comprising a triazole-based small molecule organic compound or analogue or pharmaceutically acceptable salt thereof that specifically targets mitochondrial oxidative phosphorylation of mitochondrial complex i according to any one of claims 1-3, and a pharmaceutically acceptable carrier.
6. The pharmaceutical composition of claim 5, further comprising a second agent.
7. The pharmaceutical composition according to claim 6, wherein the second agent is used for the preparation of a medicament for the prevention and/or treatment of cancer.
8. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is formulated as an injectable fluid, aerosol, cream, gel, pill, capsule, syrup, transdermal patch, or excipient.
9. The pharmaceutical composition of claim 5, further comprising an excipient, diluent, adjuvant, vehicle, or combination thereof.
10. A class of mitochondrial oxidative phosphorylation inhibitors, characterized in that it comprises a triazole class of small molecule organic compounds and pharmaceutically acceptable salts, metabolites or prodrugs which inhibit mitochondrial oxidative phosphorylation specifically targeted to mitochondrial complex i according to any one of claims 1-3.
11. Use of a triazole class small molecule organic compound that inhibits mitochondrial oxidative phosphorylation according to any one of claims 1-3, or a pharmaceutically acceptable salt, metabolite or prodrug, or a pharmaceutical composition according to claim 5, for the preparation of an oxidative phosphorylation inhibitor.
12. Use of a triazole class small molecule organic compound that specifically targets mitochondrial oxidative phosphorylation of mitochondrial complex i to inhibit mitochondrial oxidative phosphorylation of a pharmaceutically acceptable salt, metabolite or prodrug according to any one of claims 1-3, or a pharmaceutical composition according to claim 5, for the preparation of an inhibitor that inhibits oxidative phosphorylation of tumor cells at high oxidative phosphorylation levels and inhibits oxidative phosphorylation by specifically inhibiting respiratory chain complex i.
13. The use according to claim 12, wherein the specific targeted mitochondrial complex i inhibits mitochondrial oxidative phosphorylation of small-molecule organic compounds of the triazole class and pharmaceutically acceptable salts, metabolites or prodrugs, or pharmaceutical compositions, are used for inhibiting proliferation, growth, migration, infiltration, clonogenic and metastasis of tumor cells, promoting apoptosis of tumor cells, promoting autophagy of tumor cells, and/or prolonging survival of tumor patients.
14. The use according to claim 12, wherein the neoplasm comprises lung cancer, pancreatic cancer, colorectal cancer, ovarian cancer, breast cancer and acute myeloid leukemia.
15. A method for preventing and/or treating a disease associated with abnormal mitochondrial oxidative phosphorylation, characterized in that an effective amount of the triazole-based small molecule organic compound that inhibits mitochondrial oxidative phosphorylation and a pharmaceutically acceptable salt, metabolite or prodrug thereof according to any one of claims 1-3, or the pharmaceutical composition according to claim 5 or the mitochondrial oxidative phosphorylation inhibitor according to claim 10 is administered to a subject in need thereof.
CN202211389870.3A 2022-11-08 2022-11-08 Triazole small molecular organic compound for inhibiting mitochondrial oxidative phosphorylation by specific targeting complex I and application thereof Pending CN118005608A (en)

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