WO2023231948A1 - Isr抑制剂及其制备方法和应用 - Google Patents

Isr抑制剂及其制备方法和应用 Download PDF

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WO2023231948A1
WO2023231948A1 PCT/CN2023/096760 CN2023096760W WO2023231948A1 WO 2023231948 A1 WO2023231948 A1 WO 2023231948A1 CN 2023096760 W CN2023096760 W CN 2023096760W WO 2023231948 A1 WO2023231948 A1 WO 2023231948A1
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synthesis
yield
nmr
white solid
referring
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PCT/CN2023/096760
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French (fr)
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张惠斌
周金培
王力勋
裴艺峰
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中国药科大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4196—1,2,4-Triazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4245—Oxadiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • C—CHEMISTRY; METALLURGY
    • 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
    • C07D401/04—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 directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings

Definitions

  • the invention belongs to the field of biomedicine technology, and specifically relates to a class of compounds with ISR inhibitory activity and their preparation methods and applications.
  • ISR Integrated stress response
  • Activation of the ISR is a common factor in many neurodegenerative diseases. Activation of the ISR has been detected in a wide range of brain diseases, including Alzheimer's disease, Parkinson's disease, Autistic Spectrum Disorder (ASD), amyotrophic lateral sclerosis (ALS), traumatic brain disease Damage etc.
  • ASD Autistic Spectrum Disorder
  • ALS amyotrophic lateral sclerosis
  • ISR also has an impact on other diseases. Mutations in ADAR1 can cause immune diseases (including Aicardi-Goutines syndrome). Pharmacological inhibition of ISR can increase the median survival rate of ADAR mutant mice by three times and return them to normal weight. Another study showed that targeted inhibition of ISR can regulate the M1/M2 phenotype of microglia, exert a new anti-inflammatory effect on microglia, and is a potential therapeutic target for neuroinflammation after surgical brain injury. By inhibiting ISR, it can also reduce pulmonary fibrosis, treat spinal cord injury and ischemic diseases (myocardial infarction, stroke, etc.).
  • ISR signaling can regulate the concentration of eIF2 ternary complex (TC) in cells.
  • TC is a heterotrimer composed of the eukaryotic translation initiation factor eIF2, GTP, and charged methionyl-initiating transfer RNA (Met-tRNAi). Translation initiation is a multi-step process.
  • the mRNA-protein complex (mRNP) recruits a multifactor complex (MFC) containing the initiation factors eIF1, eIF3 and eIF5 and the eIF2 complex that binds GTP and initiation tRNA. Promotes the delivery of ternary complexes to the 40S ribosomal subunit.
  • MFC multifactor complex
  • MFC is most commonly recruited to mRNA by the eIF4F complex, which recognizes and binds to the 50cap structure and positions MFC to scan from 50end to the AUG start codon.
  • GTP hydrolysis by eIF2 brings 40S ribosomes into translation initiation and leads to the recruitment of the 60S subunit and the elongation phase of translation initiation.
  • GDP is exchanged for GTP by the guanine nucleotide-exchange factor (GEF) eIF2B, preparing the eIF2 complex for the next round of initiation.
  • GEF guanine nucleotide-exchange factor
  • eIF2B activators can promote the formation of eIF2B decamer protein, reverse the effects of ISR, promote translation initiation and restore protein synthesis.
  • the object of the present invention is to provide a class of compounds with ISR inhibitory activity, as well as preparation methods and applications thereof.
  • the present invention provides compounds having the structure of formula (I) or formula (II), their stereoisomers, their geometric isomers, their tautomers, their pharmaceutically acceptable salts, their prodrugs, and their hydrates. substance or its solvate,
  • A is a halogen-substituted biaromatic heterocycle
  • W is a five-membered heterocyclic ring
  • n 0, 1 or 2;
  • n 0 or 1
  • B is a halogen-substituted benzene ring
  • G is selected from
  • A is selected from
  • W is selected from
  • the present invention provides the following compounds, their stereoisomers, their geometric isomers, their tautomers Isomers, pharmaceutically acceptable salts thereof, prodrugs thereof, hydrates or solvates thereof, but are not limited to the following compounds:
  • the pharmaceutically acceptable salts include acid addition salts formed by compounds having the structure of formula (I) or formula (II) and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid , acetic acid, trifluoroacetic acid, lactic acid, oxalic acid, adipic acid, glutaric acid, malonic acid, maleic acid, succinic acid, fumaric acid, tartaric acid, citric acid, palmitic acid, benzoic acid, methanesulfonic acid, p- Toluenesulfonic acid, salicylic acid, phenylacetic acid, mandelic acid, in addition to acid salts of inorganic bases.
  • the present invention also provides a preparation method of the compound represented by formula (I) or formula (II):
  • the present invention also provides a pharmaceutical composition containing the compound of the present invention, its stereoisomers, its geometric isomers, its tautomers, its pharmaceutically acceptable salts, its prodrugs, and its hydrated
  • the drug or its solvate is the active ingredient or the main active ingredient, supplemented by a pharmaceutically acceptable carrier.
  • the present invention also provides the compounds of the present invention, their stereoisomers, their geometric isomers, their tautomers, and their medicines.
  • the diseases are neurodegenerative diseases, autoimmune diseases, musculoskeletal diseases, inflammatory diseases, and genetic disorders.
  • the neurodegenerative disease is selected from: cerebral ischemia, brain injury, epilepsy, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, different types of spinocerebellar ataxia, autism spectrum disorder, white matter ablative disease, intellectual disability syndrome, prion disease, Creutzfeldt-Jakob disease, stroke, chronic traumatic encephalopathy, spinal cord injury, dementia, frontotemporal dementia (FTD), depression, or social behavior Disorder;
  • the autoimmune disease is selected from: systemic lupus erythematosus, type 1 diabetes, multiple sclerosis or rheumatoid arthritis;
  • the musculoskeletal disease is selected from: myopathy, muscular dystrophy , muscle atrophy, muscle wasting or sarcopenia; preferably, the inflammatory disease is selected from: arthritis, ps
  • the compound provided by the invention can effectively combine with eIF2B, has good ATF4 inhibitory ability, and can better restore protein synthesis when stimulated by ISR. Therefore, the compounds of the present invention can serve as ISR inhibitors and be used to treat various related disorders.
  • the present invention is for clinical screening and/or preparation for the treatment and/or prevention of Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, aging, autism spectrum disorder, white matter ablation disease, dementia, Drugs for neurodegenerative diseases such as frontotemporal dementia (FTD) offer a new option.
  • FTD frontotemporal dementia
  • Step 1 Synthesis of ethyl 5-chlorobenzofuran-2-carboxylate
  • Step 3 Synthesis of trans-(4-(5-chlorobenzofuran-2-carboxamido)cyclohexyl)carbamic acid tert-butyl ester
  • Step 2 Synthesis of ethyl 2-(2-(4-chlorobenzoyl)hydrazino)-2-oxoacetate
  • Step 3 Synthesis of ethyl 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxylate
  • Step 2 trans-N-(4-(5-chlorobenzofuran-2-carboxamido)cyclohexyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2- Synthesis of formamide (I-1)
  • Step 2 Synthesis of ethyl 2-(2-(2-(4-chlorophenyl)acetyl)hydrazino)-2-oxoacetate
  • Step 3 Synthesis of ethyl 5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxylate
  • Step 2 Synthesis of ethyl 2-(2-(4-bromobenzoyl)hydrazino)-2-oxoacetate
  • Step 3 Synthesis of ethyl 5-(4-bromophenyl)-1,3,4-oxadiazole-2-carboxylate
  • Step 2 trans-5-(4-bromophenyl)-N-(4-(5-chlorobenzofuran-2-carboxamido)cyclohexyl)-1,3,4-oxadiazole-2- Synthesis of formamide (I-3)
  • Step 2 Synthesis of ethyl 2-(2-(4-chloro-3-fluorobenzoyl)hydrazino)-2-oxoacetate
  • Step 3 Synthesis of ethyl 5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazole-2-carboxylate
  • Step 2 trans-5-(4-chloro-3-fluorophenyl)-N-(4-(5-chlorobenzofuran-2-carboxamido)cyclohexyl)-1,3,4-oxadis Synthesis of azole-2-carboxamide (I-4)
  • Step 2 Synthesis of ethyl 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylate
  • Step 3 trans-5-chloro-N-(4-(((5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino) Synthesis of cyclohexyl)benzofuran-2-carboxamide (I-9)
  • Step 3 Synthesis of (1-(5-chlorobenzofuran-2-carboxamido)piperidin-4-yl)carbamic acid tert-butyl ester
  • Step 1 Synthesis of ((1-nitrosopiperidin-4-yl)methyl)carbamic acid tert-butyl ester
  • Step 5 N-((1-(5-chlorobenzofuran-2-carboxamide)piperidin-4-yl)methyl)-5-(4-chlorophenyl)-1,3,4-oxa Synthesis of diazole-2-carboxamide (I-12)
  • Step 3 N-((1-(5-chlorobenzofuran-2-carbonyl)piperidin-4-yl)methyl)-5-(4-chlorophenyl)-1,3,4-oxadis Synthesis of azole-2-carboxamide (I-17)
  • Step 1 Synthesis of tert-butyl 4-(5-chlorobenzofuran-2-carboxamido)piperidine-1-carboxylate
  • Step 1 Synthesis of 4-((5-chlorobenzofuran-2-carboxamido)methyl)piperidine-1-carboxylic acid tert-butyl ester
  • Step 3 Synthesis of trans-(4-(6-chloroquinoline-2-carboxamido)cyclohexyl)carbamic acid tert-butyl ester
  • Step 2 Synthesis of tert-butyl 4-(2-(5-chlorobenzofuran-2-carbonyl)hydrazine-1-carbonyl)piperidine-1-carboxylate
  • Step 1 Synthesis of trans-4-(2-(5-chlorobenzofuran-2-carbonyl)hydrazine-1-carbonyl)cyclohexyl)carbamic acid tert-butyl ester
  • Step 2 Synthesis of trans-4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexan-1-amine
  • Step 1 Synthesis of trans-((4-(2-(5-chlorobenzofuran-2-carbonyl)hydrazine-1-carbonyl)cyclohexyl)methyl)carbamic acid tert-butyl ester
  • Step 2 Synthesis of trans-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)methanamine
  • Step 3 trans-N-((4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)methyl)-2- Synthesis of (4-chlorophenoxy)acetamide (II-3)
  • Step 3 Synthesis of 4-(5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexane-1-amine
  • Step 1 Synthesis of 4-(2-(5-chloro-1H-indole-2-carbonyl)hydrazine-1-carbonyl)piperidine-1-carboxylic acid tert-butyl ester
  • the raw material is 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid. 473 mg of yellow-white solid is obtained by referring to the synthesis of step 3 in Example 1. The yield is 99.72%.
  • the raw materials were 5-chloro-1H-indole-2-carbohydrazide and (1r, 4r)-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid synthesized in step 1 of Example 44.
  • step 3 in Example 1 374 mg of yellow-white solid was obtained with a yield of 78.72%.
  • the raw materials were 5-chlorobenzofuran-2-carbohydrazide and (1r, 4r)-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid synthesized in step 1 of Example 39.
  • step 3 in Example 1 582 mg of white solid was obtained with a yield of 82.21%.
  • Step 2 (1r,4r)-4-((5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)methyl)cyclohexane-1 -Synthesis of amines.
  • Step 1 Synthesis of: (1r, 4r)-4-(2-(4-chlorophenoxy)acetamido)cyclohexane-1-carboxylic acid methyl ester
  • the raw material is (1r, 4r)-4-aminocyclohexane-1-carboxylic acid methyl ester, refer to the method described in step 3 in Example 1. 335 mg of white solid was obtained, with a yield of 72.24%.
  • Step 1 2-(4-chlorophenoxy)-N-((1r,4r)-4-(2-(6-chloroquinoline-2-carbonyl)hydrazine-1-carbonyl)cyclohexyl)acetamide synthesis
  • the raw materials are 2-(4-chlorophenoxy)-N-((1r, 4r)-4-(hydrazinecarbonyl)cyclohexyl)acetamide in step 2 of Example 53 and 6-chloroquinoline in step 2 of Example 29 -2-carboxylic acid.
  • step 3 the method described in step 3 in Example 1. 261 mg of white solid was obtained, with a yield of 72%.
  • Step 2 2-(4-chlorophenoxy)-N-((1r,4r)-4-(5-(6-chloroquinolin-2-yl)-1,3,4-oxadiazole-2 Synthesis of -yl)cyclohexyl)acetamide (II-11)
  • the raw materials are 6-bromoquinoline-2-carboxamide in step 3 of Example 55, 2-(4-chlorophenoxy)-N-((1r, 4r)-4-(hydrazinecarbonyl) in step 2 of Example 53 ) cyclohexyl)acetamide.
  • 6-Bromoquinoline-2-carboxamide (105 mg, 322.29 ⁇ mol), 2-(4-chlorophenoxy)-N-((1r, 4r)-4-(hydrazinecarbonyl)cyclohexyl)acetamide ( 78 mg, 322.29 ⁇ mol) was put into a sealed tube, dissolved with N,N-dimethylformamide, and reacted at 130°C for 5 hours. Extract with ethyl acetate, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The residue is purified by silica gel column chromatography to obtain 17 mg of white solid with a yield of 9.75%.
  • the raw materials were 2-(5-chloro-1H-indol-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole from Step 2 of Example 47 and Step 10 of Example 47. 1's 5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazole-2-carboxylic acid
  • the raw material is 2-(5-chloro-1H-indol-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole from Step 2 of Example 47, Step 2 of Example 47 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylic acid
  • Step 1 2-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)-2-oxo Synthesis of ethyl acetate
  • the raw materials were 2-(5-chlorobenzofuran-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole and oxalyl chloride monoethyl ester from step 3 of Example 39. Referring to the method described in step 2 of Example 3, 217 mg of white solid was obtained with a yield of 92.7%.
  • Step 2 2-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)-2-oxo Synthesis of Acetyl Hydrazide.
  • Step 1 Synthesis of benzyl (1s, 3s)-3-hydroxycyclobutane-1-carboxylate
  • Step 2 Synthesis of benzyl (1s, 3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylate
  • Benzyl (1s, 3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylate (1.21g, 4.40mmol) was dissolved in THF, and NaOH (880mg, 22mmol) was added to react overnight. The reaction was also concentrated, and the residue was extracted with dichloromethane/1 mol HCl aqueous solution. The organic phase was collected and spin-dried to obtain 793 mg of colorless liquid, with a yield of 97.94%.
  • Step 1 (1s,3s)-N'-(2-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidine Synthesis of -1-yl)-2-oxoacetyl)-3-(trifluoromethoxy)cyclobutane-1-carboxylhydrazide
  • Step 2 (4-(5-(5-(chlorofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)(5-((1s, Synthesis of 3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4oxadiazol-1-yl)methanone
  • the raw materials are 5-chlorobenzofuran-2-carboxylic acid and 4-aminopiperidine-1-carboxylic acid tert-butyl ester obtained in step 2 of Example 1.
  • Step 2 Synthesis of ethyl 2-(4-(5-chlorobenzofuran-2-carboxamide)piperidin-1-yl)-2-oxoacetate
  • Step 1 5-chloro-N-(1-(2-oxo-2-(2-((1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carbonyl)hydrazino) Synthesis of acetyl)piperidin-4-yl)benzofuran-2-carboxamide
  • the raw material is 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)benzofuran-2-carboxamide obtained in step 3 of Example 65, Example 63 (1s, 3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylic acid obtained in step 3.
  • 203 mg of white solid was obtained with a yield of 54.89%.
  • Step 2 5-chloro-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazole-2-carbonyl) Synthesis of piperidin-4-yl)benzofuran-2-carboxamide
  • the raw materials are 5-chloro-1H-indole-2-carboxylic acid and 4-aminopiperidine-1-carboxylic acid tert-butyl ester. Referring to the synthesis in step 1 of Example 65, 1.56 g of white solid was obtained. Yield 72.14%.
  • Step 2 Synthesis of ethyl 2-(4-(5-chloro-1H-indole-2-carboxamide)piperidin-1-yl)-2-oxoacetate
  • Step 1 5-chloro-N-(1-(2-oxo-2-(2-((1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carbonyl)hydrazino) Synthesis of acetyl)piperidin-4-yl)-1H-indole-2-carboxamide
  • the raw material is 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)-1H-indole-2-carboxamide obtained in step 3 of Example 67, and the implementation (1s, 3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylic acid obtained in step 3 of Example 63.
  • 925 mg of white solid was obtained with a yield of 41.11%.
  • Step 2 5-chloro-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazole-2-carbonyl) Synthesis of piperidin-4-yl)-1H-indole-2-carboxamide (I-30)
  • Step 1 Synthesis of: (1s, 3s)-3-(benzyloxy)cyclobutan-1-ol
  • Step 2 Synthesis of tert-butyl 2-(3-p-(benzyloxy)cyclobutoxy)acetate
  • the raw materials were 3-cis-(benzyloxy)cyclobutanol and tert-butyl 2-chloroacetate obtained in step 1 of Example 63.
  • 3-cis-(benzyloxy) ring Butanol (1.30g, 7.29mmol) and CsCO3 (2.81g, 14.59mmol) were dissolved in DMF (10ml).
  • 2-chloroacetate tert-butyl ester (1.10g, 7.29mmol) was added and the reaction was carried out overnight. Extract with ethyl acetate, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure.
  • Step 3 Synthesis of tert-butyl 2-(3-cis-hydroxycyclobutoxy)acetate
  • Step 4 Synthesis of tert-butyl 2-(3-chloro-(trifluoromethoxy)cyclobutoxy)acetate
  • the raw material is (1r, 4r)-4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexane obtained in step 2 of Example 40 -1-amine, 2-((1s,3s)-3-(trifluoromethoxy)cyclobutoxy)acetic acid obtained in step 5 of Example 69.
  • Step 3 of Example 1 34 mg of white solid was obtained with a yield of 41.22%.
  • Step 2 Synthesis of ethyl 2-(4-(6-bromoquinoline-3-carboxamide)piperidin-1-yl)-2-oxoacetate
  • Step 1 6-bromo-N-(1-(2-oxo-2-(2-((1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carbonyl)hydrazino) Synthesis of acetyl)piperidin-4-yl)quinoline-3-carboxamide
  • Step 2 6-bromo-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazole-2-carbonyl) Synthesis of piperidin-4-yl)quinoline-3-carboxamide
  • Step 1 2-(trifluoromethoxy)ethyl-2-(2-(4-(6-bromoquinoline-3-carboxamide)piperidin-1-yl)-2-oxoacetyl) Synthesis of hydrazine-1-carboxylate
  • Step 2 6-bromo-N-(1-(5-(2-(trifluoromethoxy)ethoxy)-1,3,4-oxadiazole-2-carbonyl)piperidin-4-yl )Synthesis of quinoline-3-carboxamide
  • Step 1 2-(trifluoromethoxy)ethyl-2-(2-(4-(5-chlorobenzofuran-2-carboxamide)piperidin-1-yl)-2-oxoacetyl )Synthesis of hydrazine-1-carboxylate
  • the raw materials are 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylic acid obtained in step 3 of Example 11, and 5-chloro-N-(piperidine) obtained in step 1 of Example 67. -4-yl)-1H-indole-2-carboxamide. Referring to the synthesis in step 1 of Example 66, 47 mg of white solid was obtained with a yield of 54.71%.
  • HEK-293T cells in the logarithmic growth phase were seeded into a 12-well plate and cultured for 12 hours. After adding the compound to be tested (500 nM) for 1 hour, thapsigargin (Tg, 100 nM) was added and incubated together for 3 hours. Discard the original culture medium, wash 3 times with PBS buffer solution, add cell lysis solution on ice, and collect protein samples for SDS-PAGE electrophoresis. and then transferred to NC membrane. After blocking for 1 hour at room temperature, the ATF4 primary antibody solution (1:1000) was incubated overnight at 4°C. Wash three times with TBST solution and incubate with HRP-labeled secondary antibody solution at room temperature for 1 hour. After washing with TBST solution three times, the chemiluminescence device was exposed. The gray value of the strips was analyzed using Image J software.
  • the compounds of the present invention significantly inhibit the expression of ATF4 in HEK-293T cells.
  • compounds I-13, I-14, I-18, I-19, II-2 and II-4 have slightly better or roughly equivalent inhibitory activity.
  • compounds I-14, I-18 and II-2 have better ATF4IC 50 values and show good activity. This shows that the compound has the ability to activate eIF2B, reverse nerve damage caused by ISR, and treat neurodegenerative diseases.
  • CHO cells in the logarithmic growth phase were inoculated into a 6-well plate and cultured for 12 hours. The original culture medium was discarded and the cells were washed once with PBS solution. Serum-free medium containing the compound to be tested (500 nM) was added and incubated for 1 hour. Thapsigargin (Tg, 100nM) was given and incubated for 24h.
  • the compounds of the present invention have a good promoting effect on CHO cell protein translation.
  • compounds I-2, I-9, I-10, I-11, I-15, I-16, I-17, I-25, I-26, I-28 and II-5 It can achieve a better protein translation recovery rate, and can approach or exceed the value of the blank group, indicating that the compound can significantly promote the recovery of protein synthesis by activating eIF2B, reverse nerve damage caused by ISR, and treat neurodegenerative diseases.

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Abstract

本发明公开了一类ISR抑制剂及其制备方法和应用，属于生物医药技术领域。本发明的化合物为具有式(I)或式(II)结构的化合物，其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，本发明提供的化合物，能够与eIF2B有效结合，具有良好的ATF4抑制能力，在ISR刺激时能够较好的恢复蛋白质的合成。所以，本发明的化合物可作为ISR抑制剂并用于治疗各种相关病症。

Description

ISR抑制剂及其制备方法和应用 技术领域
本发明属于生物医药技术领域，具体涉及一类具有ISR抑制活性的化合物及其制备方法和应用。
背景技术
整合应激反应(integrated stress response,ISR)是蛋白质稳态失调时诱发的信号中心调控网络，主要以控制蛋白质合成速率来实现，帮助细胞、组织和生物体适应变化的环境并维持健康。ISR的生物学功能是在压力下恢复细胞内稳态，错误折叠蛋白质的积累会激活ISR反应。该途径在翻译起始水平上起作用抑制整体蛋白质合成，同时促进参与细胞恢复的几种蛋白质的翻译。
ISR的激活是许多神经退行性疾病的共同因素。在广泛的脑部疾病中，检测到了ISR的激活，包括阿尔茨海默病、帕金森病、孤独症谱系障碍(Autistic Spectrum Disorder,ASD)、肌萎缩侧索硬化症(ALS)、创伤性脑损伤等。
ISR在其他疾病方面也具有影响作用。ADAR1的突变能够引起免疫疾病(包括Aicardi-Goutières综合征)，药理学抑制ISR能够让ADAR突变小鼠中位存活率提高3倍，并使其恢复到正常体重。另一项研究表明，靶向抑制ISR可以调节小胶质细胞M1/M2表型，对小胶质细胞发挥新的抗炎作用，是外科脑损伤后神经炎症的潜在治疗靶点。通过抑制ISR还可以达到减轻肺纤维化、治疗脊髓损伤和缺血性疾病(心肌梗死和中风等)的效果。
ISR信号能够调控细胞内eIF2三元复合物(ternary complex,TC)的浓度。TC是一种异源三聚体，是由真核翻译起始因子eIF2、GTP和带电荷甲硫酰基-起始转运RNA(Met-tRNAi)组成。翻译起始是一个多步骤过程，mRNA-蛋白质复合物(mRNP)招募包含起始因子eIF1、eIF3和eIF5以及结合GTP和起始tRNA的eIF2复合物的多因子复合物(Multifactor complex,MFC)，促进三元复合物向40S核糖体亚基的传递。MFC最常被eIF4F复合物招募到mRNA中，eIF4F复合物识别并结合到50cap结构，并定位MFC以从50end扫描到AUG起始密码子。一旦识别到AUG，eIF2对GTP的水解使40S核糖体进入翻译起始，并导致60S亚单位的招募和翻译起始的延长阶段。从核糖体释放eIF2–GDP后，GDP被鸟嘌呤核苷酸交换因子(guanine nucleotide-exchange factor,GEF)eIF2B交换为GTP，为下一轮启动准备eIF2复合物。eIF2B的活性由其底物eIF2的磷酸化、核苷酸和辅因子与eIF2B的结合以及eIF2B自身的磷酸化调节。eIF2B激活剂能促进eIF2B十聚体蛋白的形成，逆转ISR的作用，推动翻译起始并且恢复蛋白质的合成。
因此，研究具有ISR抑制活性的新型化合物对于多种疾病的治疗具有重要意义。
发明内容
本发明的目的是提供一类具有ISR抑制活性的化合物，及其制备方法和应用。
为了实现上述目的，本发明采用以下技术方案：
本发明提供具有式(I)或式(II)结构的化合物，其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，
其中：
A为卤素取代的双芳香杂环；
D选自
L为
W为五元杂环；
n为0、1或2；
m为0或1；
B为卤素取代的苯环、
K选自
G选自
在一些实施例中，
A选自
W选自
B选自
在一些具体实施例中，本发明提供以下化合物，其立体异构体、其几何异构体、其互变 异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，但不局限于以下化合物范围：


在本发明中，所述药学上可接受的盐包括具有式(I)或式(II)结构的化合物与下列酸形成的酸加成盐：盐酸、氢溴酸、硫酸、硝酸、磷酸、甲酸、乙酸、三氟乙酸、乳酸、草酸、己二酸、戊二酸、丙二酸、马来酸、琥珀酸、富马酸、酒石酸、柠檬酸、棕榈酸、苯甲酸、甲磺酸、对甲苯磺酸、水杨酸、苯基乙酸、杏仁酸，此外还包括无机碱的酸式盐。
本发明还提供式(I)或式(II)所示化合物的制备方法：
方法1：在缩合剂的作用下，化合物(III)和化合物(V)发生缩合反应，制备得到式(I)化合物；
方法2：极性非质子溶剂中，在碳酸铯的作用下，化合物(III)和化合物(V)发生取代反应，制备得到式(I)化合物；
方法3：在缩合剂的作用下，化合物(VI)和化合物(VII)发生缩合反应，制备得到式(II)化合物；
其中，以上A、B、D、L、W、K、G、m、n的定义如前所述。
本发明还提供一种药物组合物，以本发明所述化合物、其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物为活性成分或者主要活性成分，辅以药学上可接受的载体组成。
本发明还提供本发明所述化合物、其立体异构体、其几何异构体、其互变异构体、其药 学上可接受的盐、其前药、其水合物或其溶剂化物在制备治疗和/或预防ISR介导的疾病的药物中的应用。
所述疾病为神经退行性疾病、自身免疫性疾病、肌肉骨骼疾病、炎症性疾病、遗传病症。优选地，所述神经退行性疾病选自：脑缺血、脑损伤、癫痫、阿尔茨海默病、帕金森病、亨廷顿病、肌萎缩性侧索硬化、不同类型脊髓小脑共济失调、孤独症谱系障碍、白质消融性疾病、智力障碍综合征、朊病毒疾病、克雅氏病、中风、慢性创伤性脑病变、脊髓损伤、痴呆、额颞叶型痴呆(FTD)、抑郁症或社交行为障碍；优选地，所述自身免疫性疾病选自：全身性红斑狼疮、1型糖尿病、多发性硬化或类风湿性关节炎；优选地，所述肌肉骨骼疾病选自：肌病、肌肉营养不良、肌肉萎缩、肌肉消瘦或肌肉减少症；优选地，所述炎症性疾病选自：关节炎、牛皮癣性关节炎、牛皮癣、幼年型特发性关节炎、哮喘、过敏性哮喘、支气管哮喘、肺结核、慢性气管病症、囊肿性纤维化、丝球体肾炎、膜性肾病、结节病、血管炎、鱼鳞病、移植物排斥反应、间质性膀胱炎、异位性皮炎或炎症性肠病；优选地，所述遗传病症选自：唐氏综合征或MEHMO综合征(智力迟钝、癫痫发作、性腺机能减退、小头畸形和肥胖症)。
本发明提供的化合物，能够与eIF2B有效结合，具有良好的ATF4抑制能力，在ISR刺激时能够较好的恢复蛋白质的合成。所以，本发明的化合物可作为ISR抑制剂并用于治疗各种相关病症。本发明为在临床上筛选和/或制备治疗和/或预防阿尔茨海默病、帕金森病、亨廷顿病、肌萎缩性侧索硬化、衰老、孤独症谱系障碍、白质消融性疾病、痴呆、额颞叶型痴呆(FTD)等神经退行性疾病的药物提供了一种新的选择。
具体实施方式
下面结合具体实施例对本发明作进一步详细说明，但不应理解为对本发明的限制。在不背离本发明精神和实质的情况下，对本发明方法、步骤或条件所作的修改或替换，均属于本发明的范围。实施例中未注明具体条件的实验方法及未说明配方的试剂均为按照本领域常规条件。
实施例1
中间体反-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)氨基甲酸叔丁酯的合成
步骤1：5-氯苯并呋喃-2-甲酸乙酯的合成
将5-氯水杨醛(0.2g，1.28mmol)溶于N,N-二甲基甲酰胺(10mL)中，加入溴乙酸乙酯(0.32g，1.92mmol)和碳酸钾(0.41g，3.19mmol)，升温至120℃，反应10h。乙酸乙酯萃取，无水硫酸钠干燥，经减压浓缩，残留物通过硅胶柱层析(石油醚/乙酸乙酯＝200/1)纯化得白色固体204mg，产率71.09％。1H NMR(300MHz,DMSO-d6)δ(ppm):7.89(dd,J＝2.2,0.6Hz,1H),7.80(dt,J＝8.9,0.8Hz,1H),7.73(d,J＝0.9Hz,1H),7.56(dd,J＝8.9,2.3Hz,1H),4.37(q,J＝7.1Hz,2H),1.34(t,J＝7.1Hz,3H).
步骤2：5-氯苯并呋喃-2-羧酸的合成
将5-氯苯并呋喃-2-甲酸乙酯(3.15g，14.00mmol)溶于四氢呋喃/乙醇(15mL/10mL)中,加入氢氧化锂(503mg，21.00mmol)，室温反应。反应5h。在冰浴条件下，用1mol/L的盐酸调pH至3，过滤，滤饼经水洗涤和少量乙酸乙酯洗涤，干燥，得到白色固体2.29g，产率83.02％。1H NMR(300MHz,DMSO-d6)δ(ppm):13.78(s,1H),7.87(d,J＝2.2Hz,1H),7.77(d,J＝8.8Hz,1H),7.63(d,J＝0.9Hz,1H),7.53(dd,J＝8.9,2.2Hz,1H).
步骤3：反-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)氨基甲酸叔丁酯的合成
将5-氯苯并呋喃-2-羧酸(0.60g，3.05mmol)溶于15mL的N,N-二甲基甲酰胺中，依次加入HATU(2.32g，6.10mmol)、N-Boc-反式-1,4-环己二胺(0.65mg，3.05mmol)和DIPEA(1.58g，12.21mmol)，室温下搅拌反应。反应3h。加入30mL水，过滤，滤饼用少量二氯甲烷洗涤，干燥，得到白色固体0.948g，产率79.06％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.62(d,J＝8.1Hz,1H),7.87(d,J＝2.2Hz,1H),7.70(d,J＝8.8Hz,1H),7.56-7.45(m,2H),6.79(d,J＝8.0Hz,1H),3.79-3.63(m,1H),3.20(d,J＝10.6Hz,1H),1.81(d,J＝11.5Hz,4H),1.48(d,J＝12.5Hz,2H),1.38(s,9H),1.33-1.20(m,2H).。
实施例2
中间体反-N-(4-氨基环己基)-5-氯苯并呋喃-2-甲酰胺的合成
将反-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)氨基甲酸叔丁酯(948mg，2.41mmol)溶于二氯甲烷(10mL)中，加入三氟乙酸(1mL)，室温条件下反应8h。将反应液减压浓缩，用氨水调pH至10，过滤，滤饼洗涤，干燥，得到淡黄色固体586mg，产率82.95％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.68(d,J＝8.0Hz,1H),7.88(d,J＝2.7Hz,1H),7.70(d,J＝8.8Hz,1H),7.55-7.45(m,2H),3.75(s,1H),2.99(d,J＝8.4Hz,1H),2.01-1.85(m,4H),1.42(qd,J＝11.4,5.6Hz,4H).。
实施例3
5-(4-氯苯基)-1,3,4-噁二唑-2-甲酸乙酯的合成
步骤1：4-氯苯甲酰肼的合成
将4-氯苯甲酸乙酯(5.00g，27.08mmol)溶于甲醇(50mL)中，加入80％水合肼(6.78g，135.41mmol)，升温至80℃回流反应，反应12h。将反应液浓缩，用少量石油醚/乙酸乙酯洗涤，过滤，干燥，得到白色固体4.10g，产率88.65％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.73(s,1H),7.73-7.66(m,2H),7.48-7.41(m,2H),5.87(s,2H).
步骤2：2-(2-(4-氯苯甲酰基)肼基)-2-氧代乙酸乙酯的合成
将4-氯苯甲酰肼(1.96g，11.51mmol)溶于二氯甲烷(15mL)中，加入三乙胺(3.20mL，23.03mmol)，接着在冰浴条件下，缓慢滴加草酰氯单乙酯(3.14g，23.03mmol)，滴加完毕撤去冰浴，室温反应1h。将反应液减压浓缩，残留物通过硅胶柱层析(石油醚/乙酸乙酯＝3/1～1/1)纯化得白色固体2.36g，产率75.74％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.98(s,1H),10.70(s,1H),7.94-7.87(m,2H),7.64-7.58(m,2H),4.22(q,J＝7.1Hz,2H),1.31(t,J＝7.1Hz,3H).
步骤3：5-(4-氯苯基)-1,3,4-噁二唑-2-甲酸乙酯的合成
将2-(2-(4-氯苯甲酰基)肼基)-2-氧代乙酸乙酯(2.11g，7.80mmol)溶于乙腈(15mL)中，缓慢加入三氯氧磷(3.59g，23.39mmol)，升温至85℃封管反应8h。冰浴条件下缓慢加入水，然后用乙酸乙酯萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，残留物通过硅胶柱层析(石油醚/乙酸乙酯＝80/1)纯化得白色固体807mg，产率40.97％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.13-8.05(m,2H),7.77-7.69(m,2H),4.47(q,J＝7.1Hz,2H),1.38(t,J＝7.1Hz,3H).。
实施例4
反-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-1)的合成
步骤1：5-(4-氯苯基)-1,3,4-噁二唑-2-羧酸的合成
参照实施例1中步骤2所述的方法，得到白色固体149mg，产率57.91％。1H NMR(300MHz,Chloroform-d)δ(ppm):8.50(s,1H),8.08-8.02(m,2H),7.57-7.50(m,2H).
步骤2：反-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-1)的合成
将5-(4-氯苯基)-1,3,4-噁二唑-2-羧酸(59mg，263μmol)溶于乙腈(10mL)中，在冰浴条件下加入草酰氯(25μL，289μmol)，再加入1滴N,N-二甲基甲酰胺，反应30min。将反-N-(4-氨基环己基)-5-氯苯并呋喃-2-甲酰胺(77mg，263μmol)溶于二氯甲烷(5mL)中，加入N,N-二异丙基乙胺(102mg，784μmol)，随后在冰浴条件下缓慢加入上述反应液，加入完毕后撤去冰浴，室温反应。反应3h。用二氯甲烷萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，残留物通过硅胶柱层析(二氯甲烷/甲醇＝200/1)纯化得到白色固体75mg，产率57.11％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.31(d,J＝8.2Hz,1H),8.68(d,J＝8.1Hz,1H),8.13(d,J＝2.0Hz,1H),8.10(d,J＝2.0Hz,1H),7.88(d,J＝2.2Hz,1H),7.76-7.68(m,3H),7.54(d,J＝1.0Hz,1H),7.49(dd,J＝8.8,2.2Hz,1H),3.79(d,J＝9.1Hz,2H),1.91(d,J＝9.4Hz,4H),1.56(q,J＝9.5Hz,4H)；MS(ESI)m/z:calculated for C24H20Cl2N4O4[M+H]+:499.09,found:499.0926.。
实施例5
中间体5-(4-氯苄基)-1,3,4-噁二唑-2-甲酸乙酯的合成
步骤1：2-(4-氯苯基)乙酰肼的合成
参照实施例3中步骤1所述的方法，得到白色固体192mg，产率96.00％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.23(s,1H),7.38-7.33(m,2H),7.30-7.25(m,2H),4.22(d,J＝3.2Hz,2H),3.35(s,2H).
步骤2：2-(2-(2-(4-氯苯基)乙酰基)肼基)-2-氧代乙酸乙酯的合成
参照实施例3中步骤2所述的方法，得到白色固体1.29g，产率68.30％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.81(s,1H),10.30(s,1H),7.42-7.36(m,2H),7.35-7.29(m,2H),4.26(q,J＝7.1Hz,2H),3.52(s,2H),1.28(t,J＝7.1Hz,3H).
步骤3：5-(4-氯苄基)-1,3,4-噁二唑-2-甲酸乙酯的合成
参照实施例3中步骤3所述的方法，得到白色固体821mg，产率68.47％。1H NMR(300MHz,Chloroform-d)δ7.37-7.33(m,2H),7.31-7.27(m,3H),4.51(q,J＝7.1Hz,2H),4.28(s,2H),1.45(t,J＝7.1Hz,3H).。
实施例6
反-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-5-(4-氯苄基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-2)的合成
步骤1：5-(4-氯苄基)-1,3,4-噁二唑-2-羧酸的合成
参照实施例1中步骤2所述的方法，得到白色固体138mg，产率78.23％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.25(s,1H),7.41-7.35(m,2H),7.35-7.28(m,2H).
步骤2：反-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-5-(4-氯苄基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-2)的合成
参照实施例4中步骤2所述的方法，得到白色固体48mg，产率55.78％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.23(d,J＝8.2Hz,1H),8.66(d,J＝8.1Hz,1H),7.87(d,J＝2.2Hz,1H),7.70(d,J＝8.9Hz,1H),7.53(d,J＝0.9Hz,1H),7.49(dd,J＝8.9,2.2Hz,1H),7.47-7.42(m,2H),7.42-7.35(m,2H),4.39(s,2H),3.75(s,2H),1.86(t,J＝10.9Hz,4H),1.52(q,J＝11.3Hz,4H)；MS(ESI)m/z:calculated for C25H22Cl2N4O4[M+H]+:513.10,found:513.3.。
实施例7
中间体5-(4-溴苯基)-1,3,4-噁二唑-2-甲酸乙酯的合成
步骤1：4-溴苯甲酰肼的合成
参照实施例3中步骤1所述的方法，得到白色固体267mg，产率53.40％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.87(s,1H),7.79-7.74(m,2H),7.69-7.64(m,2H),4.53(s,2H).
步骤2：2-(2-(4-溴苯甲酰基)肼基)-2-氧代乙酸乙酯的合成
参照实施例3中步骤2所述的方法，得到白色固体252mg，产率85.99％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.99(s,1H),10.75-10.68(m,1H),7.85-7.80(m,2H),7.78-7.73(m,2H),4.31(q,J＝7.1Hz,2H),1.31(t,J＝7.1Hz,3H).
步骤3：5-(4-溴苯基)-1,3,4-噁二唑-2-甲酸乙酯的合成
参照实施例3中步骤3所述的方法，得到白色固体153mg，产率65.70％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.05-7.98(m,2H),7.90-7.84(m,2H),4.46(q,J＝7.1Hz,2H),1.37(t,J＝7.1Hz,3H).。
实施例8
反-5-(4-溴苯基)-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-3)的合成
步骤1：5-(4-溴苯基)-1,3,4-噁二唑-2-羧酸的合成
参照实施例1中步骤2所述的方法，得到白色固体102mg，产率73.62％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.39(s,1H),8.00-7.95(m,2H),7.86-7.81(m,2H).
步骤2：反-5-(4-溴苯基)-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-3)的合成
参照实施例4中步骤2所述的方法，得到白色固体85mg，产率27.31％。1H NMR(300MHz,DMSO-d6)δ(ppm)9.30(d,J＝8.2Hz,1H),8.68(d,J＝8.1Hz,1H),8.09-8.00(m,2H),7.93-7.83(m,3H),7.71(d,J＝8.8Hz,1H),7.54(d,J＝0.9Hz,1H),7.49(dd,J＝8.8,2.2Hz,1H),3.79(d,J＝8.3Hz,2H),1.91(d,J＝9.1Hz,4H),1.56(q,J＝9.7Hz,4H)；MS(ESI)m/z:calculated for C24H20BrClN4O4[M+H]+:543.04,found:543.0428.。
实施例9
中间体5-(4-氯-3-氟苯基)-1,3,4-噁二唑-2-甲酸乙酯的合成
步骤1：3-氟-4-氯苯甲酰肼的合成
参照实施例3中步骤1所述的方法，得到白色固体1.73g，产率91.88％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.95(s,1H),7.85-7.76(m,1H),7.75-7.66(m,2H),4.47(s,2H).
步骤2：2-(2-(4-氯-3-氟苯甲酰基)肼基)-2-氧代乙酸乙酯的合成
参照实施例3中步骤2所述的方法，得到白色固体540mg，产率69.86％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.91(s,2H),7.90-7.84(m,1H),7.83-7.76(m,2H),4.31(q,J＝7.1Hz,2H),1.31(t,J＝7.1Hz,3H).
步骤3：5-(4-氯-3-氟苯基)-1,3,4-噁二唑-2-甲酸乙酯的合成
参照实施例3中步骤3所述的方法，得到白色固体376mg，产率74.26％。1H NMR(300MHz,Chloroform-d)δ(ppm):8.00-7.92(m,2H),7.66-7.59(m,1H),4.58(q,J＝7.2Hz,2H),1.51(t,J＝7.1Hz,3H).。
实施例10
反-5-(4-氯-3-氟苯基)-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-4)的合成
步骤1：5-(4-氯-3-氟苯基)-1,3,4-噁二唑-2-羧酸的合成
参照实施例1中步骤2所述的方法，得到白色固体243mg，产率70.38％。1H NMR(300MHz,Methanol-d4)δ(ppm):8.05-7.94(m,2H),7.74(dd,J＝8.4,7.4Hz,1H).
步骤2：反-5-(4-氯-3-氟苯基)-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-4)的合成
参照实施例4中步骤2所述的方法，得到白色固体104mg，产率29.43％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.28(d,J＝8.1Hz,1H),8.65(d,J＝8.1Hz,1H),8.11(dd,J＝9.6,1.9Hz,1H),7.97(dd,J＝8.5,1.9Hz,1H),7.93-7.86(m,2H),7.71(d,J＝8.8Hz,1H),7.54(d,J＝0.9Hz,1H),7.49(dd,J＝8.9,2.2Hz,1H),3.81(s,2H),1.92(d,J＝8.9Hz,4H),1.56(q,J＝9.8Hz,4H)；MS(ESI)m/z:calculated for C24H19ClFN4O4[M+H]+:517.08,found:517.1.。
实施例11
中间体3-(4-氯苯基)-1,2,4-噁二唑-5-羧酸的合成
步骤1：(Z)-4-氯-N'-羟基苯甲酰胺的合成
将4-氯苯腈(0.30g，2.18mmol)溶于甲醇(10mL)中，盐酸羟胺(227mg，3.27mmol)和碳酸钠(173mg，1.64mmol)溶于水(1mL)并加入4-氯苯腈的甲醇溶液中，升温至65℃反应15h。将反应液减压浓缩，乙酸乙酯萃取，无水硫酸钠干燥，经减压浓缩，得到粗品黄色固体337mg，产率90.58％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.85(s,1H),7.88-7.80(m,2H),7.58-7.50(m,2H),4.51(s,2H).
步骤2：3-(4-氯苯基)-1,2,4-噁二唑-5-甲酸乙酯的合成
将(Z)-4-氯-N'-羟基苯甲酰胺(337mg，1.98mmol)溶于四氢呋喃中，加入N,N-二异丙基乙 胺(412μl，2.96mmol)，缓慢加入草酰氯单乙酯(405mg，2.96mmol)，升温至70℃，反应2h。将反应液减压浓缩，乙酸乙酯萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，残留物通过硅胶柱层析(石油醚/乙酸乙酯＝150/1～50/1)纯化得白色固体331mg，产率66.32％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.13-8.05(m,2H),7.77-7.64(m,2H),4.47(q,J＝7.1Hz,2H),1.38(t,J＝7.1Hz,3H).
步骤3：3-(4-氯苯基)-1,2,4-噁二唑-5-羧酸的合成
参照实施例1中步骤2所述的方法，得到白色固体147mg，产率49.96％。1H NMR(300MHz,Methanol-d4)δ(ppm):8.15-8.09(m,2H),7.60-7.54(m,2H).。
实施例12
反-N-(4-(5-氯苯并呋喃-2-甲酰胺基)环己基)-3-(4-氯苯基)-1,2,4-噁二唑-5-甲酰胺(Ⅰ-5)的合成
参照实施例4中步骤2所述的方法，得到白色固体107mg，产率42.39％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.42(d,J＝8.2Hz,1H),8.66(d,J＝8.1Hz,1H),8.13-8.06(m,2H),7.88(d,J＝2.2Hz,1H),7.70(dt,J＝9.2,2.4Hz,3H),7.54(d,J＝0.9Hz,1H),7.49(dd,J＝8.8,2.2Hz,1H),3.87-3.74(m,2H),1.96-1.85(m,4H),1.56(q,J＝9.7Hz,4H)；MS(ESI)m/z:calculated for C24H20Cl2N4O4[M+H]+:499.09,found:499.0934.。
实施例13
反-5-氯-N-(4-(((5-(4-氯苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)苯并呋喃-2-甲酰胺(Ⅰ-6)的合成
步骤1：4-氯-N'-(2-氯乙酰基)苯甲酰肼的合成
将4-氯苯甲酰肼(1.92g，11.28mmol)溶于二氯甲烷(20mL)中，加入DIPEA(2.92g，22.56mmol)，加入氯乙酰氯(1.35mL，16.92mmol)，室温下反应50min。将反应液减压浓缩，用水搅拌洗涤，过滤，滤饼用石油醚/乙酸乙酯洗涤，过滤，滤饼干燥，得到白色固体2.11g，产率75.72％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.64(s,1H),10.42(s,1H),7.90(d,J＝8.3Hz,2H),7.60(d,J＝8.2Hz,2H),4.22(s,2H).
步骤2：2-(氯甲基)-5-(4-氯苯基)-1,3,4-噁二唑的合成
将4-氯-N'-(2-氯乙酰基)苯甲酰肼(1.78g，7.20mmol)溶于乙腈(15mL)中，加入POCl3(2.01mL，21.59mmol)，封管反应并升温至85℃，反应10h。缓慢加入水，用乙酸乙酯萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，残留物通过硅胶柱层析(石油醚/乙酸乙酯＝80/1)纯化得到白色固体856mg，产率51.93％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.09-8.00(m,2H),7.77-7.67(m,2H),5.16(s,2H).
步骤3：反-5-氯-N-(4-(((5-(4-氯苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)苯并呋喃-2-甲 酰胺(Ⅰ-6)的合成
将2-(氯甲基)-5-(4-氯苯基)-1,3,4-噁二唑(132mg，450μmol)溶于N,N-二甲基甲酰胺(10mL)中，加入碳酸钾(124mg，900μmol)室温搅拌10min，接着缓慢加入N-(4-氨基环己基)-5-氯苯并呋喃-2-甲酰胺(103mg，450μmol)，室温反应4h。用乙酸乙酯萃取，无水硫酸钠干燥，经减压浓缩，残留物通过硅胶柱层析(二氯甲烷/甲醇＝100/1)纯化得到白色固体67mg，产率30.70％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.57(d,J＝8.1Hz,1H),8.05-7.99(m,2H),7.86(d,J＝2.2Hz,1H),7.70(dt,J＝8.9,2.4Hz,3H),7.51(d,J＝0.9Hz,1H),7.47(dd,J＝8.9,2.2Hz,1H),4.05(s,2H),3.81-3.70(m,1H),2.44(d,J＝10.7Hz,1H),2.00-1.91(m,2H),1.83(d,J＝12.3Hz,2H),1.47-1.34(m,2H),1.21-1.10(m,2H)；MS(ESI)m/z:calculated for C24H22Cl2N4O3[M+H]+:485.11,found:485.1133.。
实施例14
反-5-氯-N-(4-(((5-(4-氯苄基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)苯并呋喃-2-甲酰胺(Ⅰ-7)的合成
步骤1：2-氯-N'-(2-(4-氯苯基)乙酰基)乙酰肼的合成
参照实施例13中步骤1所述的方法，得到淡黄色固体710mg，产率83.67％。1H NMR(300MHz,,DMSO-d6)δ(ppm):10.38-10.29(m,2H),7.41-7.37(m,2H),7.32(d,J＝8.5Hz,2H),4.14(s,2H),3.51(s,2H).
步骤2：2-(4-氯苄基)-5-(氯甲基)-1,3,4-噁二唑的合成
参照实施例13中步骤2所述的方法，得到白色固体282mg，产率59.63％。1H NMR(300MHz,DMSO-d6)δ(ppm):7.47-7.41(m,2H),7.40-7.35(m,2H),5.03(s,2H),4.35(s,2H).
步骤3：反-5-氯-N-(4-(((5-(4-氯苄基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)苯并呋喃-2-甲酰胺(Ⅰ-7)的合成
参照实施例13中步骤3所述的方法，得到白色固体89mg，产率29.08％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.55(d,J＝8.1Hz,1H),7.85(d,J＝2.2Hz,1H),7.68(d,J＝8.8Hz,1H),7.51(d,J＝1.0Hz,1H),7.49-7.40(m,3H),7.37-7.31(m,2H),4.28(s,2H),3.90(s,2H),3.77-3.65(m,1H),2.33(d,J＝11.0Hz,1H),1.90-1.75(m,4H),1.36(q,J＝11.9Hz,2H),1.07(q,J＝11.4Hz,2H)；MS(ESI)m/z:calculated for C24H22Cl2N4O3[M+H]+:499.12,found:499.1303.。
实施例15
反-N-(4-(((5-(4-溴苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)-5-氯苯并呋喃-2-甲酰胺(Ⅰ-8)的合成
步骤1：4-溴-N'-(2-氯乙酰基)苯甲酰肼的合成
参照实施例13中步骤1所述的方法，得到白色固体163mg，产率45.03％。
步骤2：2-(4-溴苯基)-5-(氯甲基)-1,3,4-噁二唑的合成
参照实施例13中步骤2所述的方法，得到白色固体86mg，产率89.87％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.00-7.92(m,2H),7.88-7.79(m,2H),5.15(s,2H).
步骤3：反-N-(4-(((5-(4-溴苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)-5-氯苯并呋喃-2-甲酰胺(Ⅰ-8)的合成
参照实施例13中步骤3所述的方法，得到白色固体108mg，产率29.34％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.58(d,J＝8.1Hz,1H),7.99-7.92(m,2H),7.89-7.81(m,3H),7.69(d,J＝8.8Hz,1H),7.52(d,J＝0.9Hz,1H),7.47(dd,J＝8.9,2.2Hz,1H),4.05(s,2H),3.81-3.69(m,1H),2.44(d,J＝10.7Hz,1H),2.00-1.91(m,2H),1.83(d,J＝12.4Hz,2H),1.45-1.32(m,2H),1.24-1.14(m,2H)；MS(ESI)m/z:calculated for C24H22BrClN4O3[M+H]+:529.06,found:529.0640.。
实施例16
反-5-氯-N-(4-(((5-(4-氯-3-氟苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)苯并呋喃-2-甲酰胺(Ⅰ-9)的合成
步骤1：4-氯-N'-(2-氯乙酰基)-3-氟苯甲酰肼的合成
参照实施例13中步骤1所述的方法，得到白色固体613mg，产率86.19％。
步骤2：2-(4-氯-3-氟苯基)-5-(氯甲基)-1,3,4-噁二唑的合成
参照实施例13中步骤2所述的方法，得到白色固体306mg，产率53.56％。1H NMR(300MHz,Chloroform-d)δ(ppm):7.92-7.83(m,2H),7.64-7.55(m,1H),4.81(s,2H).
步骤3：反-5-氯-N-(4-(((5-(4-氯-3-氟苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)苯并呋喃-2-甲酰胺(Ⅰ-9)的合成
参照实施例13中步骤3所述的方法，得到白色固体84mg，产率27.30％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.59(d,J＝8.1Hz,1H),8.04-7.97(m,1H),7.87(dd,J＝3.7,1.4Hz,3H),7.69(d,J＝8.8Hz,1H),7.51(d,J＝0.9Hz,1H),7.47(dd,J＝8.8,2.2Hz,1H),4.06(s,2H),3.75(dt,J＝7.8,3.8Hz,1H),2.44(d,J＝10.9Hz,1H),1.96(d,J＝12.6Hz,2H),1.83(d,J＝11.9Hz,2H),1.48-1.36(m,2H),1.14(d,J＝13.0Hz,2H)；MS(ESI)m/z:calculated for C24H21Cl2FN4O3[M+H]+:503.10,found:503.2.。
实施例17
中间体(1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)氨基甲酸叔丁酯的合成
步骤1：(1-亚硝基哌啶-4-基)氨基甲酸叔丁酯的合成
将N-Boc-氨基哌啶(300mg，1.50mmol)溶于四氢呋喃(10mL)中，在冰浴条件下滴入1mol/L盐酸(2.70mL，2.70mmol)，接着缓慢加入溶于5mL水的亚硝酸钠(248mg，3.59mmol)。反应3h。用乙酸乙酯和水萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，得到白色固体315mg，产率91.72％。无需进一步处理，直接用于下一步反应。
步骤2：(1-氨基哌啶-4-基)氨基甲酸叔丁酯的合成
将(1-亚硝基哌啶-4-基)氨基甲酸叔丁酯(340mg，1.48mmol)溶于乙酸/水(1:1，15mL)中，加入锌粉(383mg，5.93mmol)，保持0℃反应。反应12h。将反应液抽滤，滤液浓缩，加入乙酸乙酯(30mL)洗涤，搅拌30min，抽滤，将滤液浓缩，干燥，得到透明油状液体251mg，产率78.62％，无需进一步处理，直接用于下一步反应。
步骤3：(1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)氨基甲酸叔丁酯的合成
将5-氯苯并呋喃-2-羧酸(114mg，0.58mmol)溶于N,N-二甲基甲酰胺(5mL)，依次加入HATU(331mg，0.87mmol)和N,N-二异丙基乙胺(0.30mL，1.74mmol)，室温搅拌15min后，接着加入(1-氨基哌啶-4-基)氨基甲酸叔丁酯(125mg，0.58mmol)。反应10h。向反应液中加入水，搅拌10min，抽滤，用少量乙酸乙酯洗涤，得到白色固体123mg，产率53.79％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.77(s,1H),7.95(s,1H),7.86(d,J＝2.2Hz,1H),7.70(d,J＝8.8Hz,1H),7.53-7.45(m,2H),6.85(d,J＝7.8Hz,1H),2.96(d,J＝10.5Hz,2H),2.78-2.66(m,4H),1.75(d,J＝12.5Hz,2H),1.61-1.47(m,2H),1.39(s,9H).。
实施例18
N-(1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(I-10)的合成
步骤1：N-(4-氨基哌啶-1-基)-5-氯苯并呋喃-2-甲酰胺的合成
参照实施例2中所述的方法，得到淡黄色固体50mg，产率50.54％。1H NMR(300MHz,Chloroform-d)δ(ppm):7.66(d,J＝2.0Hz,1H),7.48(d,J＝0.8Hz,1H),7.46-7.35(m,3H),3.25(dt,J＝10.6,3.9Hz,2H),2.86(dq,J＝10.2,6.3,5.2Hz,1H),2.75(td,J＝11.0,2.8Hz,2H),1.96(d,J＝3.8Hz,2H),1.75-1.67(m,2H).
步骤2：N-(1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(I-10)的合成
将5-(4-氯苯基)-1,3,4-噁二唑-2-羧酸(39mg，0.17mmol)溶于溶于N,N-二甲基甲酰胺(5mL)，依次加入HATU(84mg，0.22mmol)和N-(4-氨基哌啶-1-基)-5-氯苯并呋喃-2-甲酰胺(50mg，0.17mmol)，室温搅拌5min后，接着加入N,N-二异丙基乙胺(74μL，0.43mmol)。反应6h。用二氯甲烷萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，残留物通过硅胶柱层析(二氯甲烷/甲醇＝200/1～100/1)纯化得到白色固体21mg，产率24.66％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.84(s,1H),9.39(d,J＝8.0Hz,1H),8.11(d,J＝8.2Hz,2H),7.88(s,1H),7.72(t,J＝8.1Hz,3H),7.55-7.45(m,2H),3.85(s,1H),3.06(d,J＝10.3Hz,2H),2.83(s,2H),1.85(s,4H)；MS(ESI)m/z:calculated for C23H19Cl2N5O4[M+H]+:500.08,found:500.1.。
实施例19
N-(1-(5-氯苯并呋喃-2-甲酰胺)哌啶-4-基)-5-(4-氯苄基)-1,3,4-噁二唑-2-甲酰胺(I-11)的合成
参照实施例18中步骤2所述的方法，得到白色固体34mg，产率26.60％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.82(s,1H),9.31(d,J＝8.1Hz,1H),7.87(d,J＝2.2Hz,1H),7.70(d,J＝8.9Hz,1H),7.56-7.46(m,3H),7.46-7.42(m,2H),7.39(d,J＝8.5Hz,2H),4.39(s,2H),3.78(q,J＝7.8Hz,1H),3.02(d,J＝10.9Hz,2H),2.84-2.75(m,2H),1.80(s,4H)；MS(ESI)m/z:calculated for C24H21Cl2N5O4[M+H]+:514.10,found:514.5.。
实施例20
中间体((1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)甲基)氨基甲酸叔丁酯的合成
步骤1：((1-亚硝基哌啶-4-基)甲基)氨基甲酸叔丁酯的合成
参照实施例17中步骤1所述的方法，得到白色固体876mg，产率96.45％。无需进一步处理，直接用于下一步反应。
步骤2：((1-氨基哌啶-4-基)甲基)氨基甲酸叔丁酯的合成
参照实施例17中步骤2所述的方法，得到透明油状液体671mg，产率81.27％。无需进一步处理，直接用于下一步反应。
步骤3：((1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)甲基)氨基甲酸叔丁酯的合成
参照实施例17中步骤3所述的方法，得到白色固体151mg，产率42.45％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.73(s,1H),7.86(d,J＝2.2Hz,1H),7.70(d,J＝8.8Hz,1H),7.53-7.45(m,2H),6.87(t,J＝5.8Hz,1H),2.99(d,J＝10.4Hz,2H),2.85(t,J＝6.1Hz,2H),2.75-2.60(m,2H),1.65(d,J＝12.8Hz,2H),1.39(s,9H),1.35-1.14(m,3H).。
实施例21
N-((1-(5-氯苯并呋喃-2-甲酰胺)哌啶-4-基)甲基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(I-12)的合成
步骤1：N-(4-(氨基甲基)哌啶-1-基)-5-氯苯并呋喃-2-甲酰胺的合成
参照实施例2中所述的方法，得到白色固体97mg，产率85.70％。；MS(ESI)m/z:calculated for C15H18ClN3O2[M+H]+:308.1,found:308.3.
步骤5：N-((1-(5-氯苯并呋喃-2-甲酰胺)哌啶-4-基)甲基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(I-12)的合成
参照实施例18中步骤2所述的方法，得到白色固体41mg，产率49.07％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.76(s,1H),9.42(t,J＝6.1Hz,1H),8.14-8.08(m,2H),7.86(d,J＝2.2Hz,1H),7.75-7.68(m,3H),7.51(s,1H),7.48(dd,J＝8.9,2.3Hz,1H),3.24(t,J＝6.4Hz,2H),3.03(d,J＝10.3Hz,2H),2.70(t,J＝10.9Hz,2H),1.75(d,J＝12.9Hz,2H),1.65(s,1H),1.36(t,J＝11.3Hz,2H)；MS(ESI)m/z:calculated for C24H21Cl2N5O4[M+H]+:514.10,found:514.2.。
实施例22
N-((1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)甲基)-5-(4-氯苄基)-1,3,4-噁二唑-2-甲酰胺(I-13)的合成
参照实施例18中步骤2所述的方法，得到白色固体34mg，产率39.61％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.77(s,1H),9.34(t,J＝6.0Hz,1H),7.87(d,J＝2.3Hz,1H),7.70(d,J＝8.9Hz,1H),7.52-7.42(m,4H),7.39(d,J＝8.6Hz,2H),4.39(s,2H),3.17(t,J＝6.3Hz,2H),3.00(d,J＝10.8Hz,2H),2.76-2.61(m,2H),1.70(d,J＝13.4Hz,2H),1.60(s,1H),1.33(d,J＝14.8Hz,2H)；MS(ESI)m/z:calculated for C25H23Cl2N5O4[M+H]+:528.11,found:528.5.。
实施例23
5-(4-溴苯基)-N-((1-(5-氯苯并呋喃-2-甲酰胺基)哌啶-4-基)甲基)-1,3,4-噁二唑-2-甲酰胺(I-14)的合成
参照实施例18中步骤2所述的方法，得到白色固体48mg，产率46.38％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.78(s,1H),9.44(t,J＝6.0Hz,1H),8.08-7.99(m,2H),7.87(dt,J＝6.6,2.5Hz,3H),7.70(d,J＝8.9Hz,1H),7.52-7.45(m,2H),3.24(t,J＝6.3Hz,2H),3.02(d,J＝10.2Hz,2H),2.76-2.66(m,2H),1.75(d,J＝13.1Hz,2H),1.64(s,1H),1.41-1.31(m,2H)；MS(ESI)m/z:calculated for C24H21BrClN5O4[M+H]+:558.05,found:558.4.。
实施例24
5-(4-氯-3-氟苯基)-N-((1-(5-氯苯并呋喃-2-甲酰胺)哌啶-4-基)甲基)-1,3,4-噁二唑-2-甲酰胺(I-15)的合成
参照实施例18中步骤2所述的方法，得到白色固体30mg，产率29.40％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.78(s,1H),9.45(t,J＝6.0Hz,1H),8.11(dd,J＝9.5,1.9Hz,1H),7.96(dd,J＝8.5,1.9Hz,1H),7.91(d,J＝7.4Hz,1H),7.89-7.85(m,1H),7.70(d,J＝8.8Hz,1H),7.54-7.45(m,2H),3.24(t,J＝6.5Hz,2H),3.02(d,J＝10.3Hz,2H),2.74-2.65(m,2H),1.75(d,J＝12.9Hz,2H),1.65(s,1H),1.36(t,J＝10.8Hz,2H)；MS(ESI)m/z:calculated for C24H20Cl2FN5O4[M+H]+:532.09,found:532.4.。
实施例25
N-(1-(5-氯苯并呋喃-2-羰基)哌啶-4-基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(I-16)的合成
步骤1：(1-(5-氯苯并呋喃-2-羰基)哌啶-4-基)氨基甲酸叔丁酯的合成
将5-氯苯并呋喃-2-羧酸(200mg，1.02mmol)溶于N,N-二甲基甲酰胺(5mL)，依次加入HATU(774mg，2.03mmol)和N,N-二异丙基乙胺(0.53mL，3.05mmol)，室温搅拌15min后，接着加入N-Boc氨基哌啶(214mg，1.07mmol)，反应4.5h。用乙酸乙酯萃取，无水硫酸钠干燥，经减压浓缩，残留物通过硅胶柱层析(石油醚/乙酸乙酯＝5/1)纯化得到乳白色固体141mg，产率36.58％。1H NMR(300MHz,Chloroform-d)δ(ppm):7.64(dd,J＝2.2,0.6Hz,1H),7.47(dt,J＝8.8,0.8Hz,1H),7.37(dd,J＝8.8,2.1Hz,1H),7.23(d,J＝0.9Hz,1H),6.82(d,J＝8.1Hz,1H),4.52(s,2H),3.88-3.68(m,1H),3.35-2.92(m,2H),2.17-2.02(m,2H),1.67(d,J＝17.8Hz,2H),1.47(s,9H).
步骤2：(4-氨基哌啶-1-基)(5-氯苯并呋喃-2-基)甲酮的合成
参照实施例2所述的方法，通过硅胶柱层析(二氯甲烷/甲醇＝100/1)纯化得到白色固体62mg，产率59.77％。1H NMR(300MHz,DMSO-d6)δ(ppm):7.82(d,J＝2.2Hz,1H),7.72(d,J＝8.8Hz,1H),7.47(dd,J＝8.9,2.2Hz,1H),7.32(s,1H),4.04(s,2H),3.08-2.82(m,3H),1.84-1.75(m,2H),1.28-1.18(m,2H).
步骤3：N-(1-(5-氯苯并呋喃-2-羰基)哌啶-4-基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(Ⅱ-7)的合成
参照实施例18中步骤2所述的方法，通过硅胶柱层析(石油醚/乙酸乙酯＝2/1)纯化得到白色固体145mg，产率64.06％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.38(d,J＝8.0Hz,1H), 8.15-8.08(m,2H),7.84(d,J＝2.2Hz,1H),7.73(dd,J＝8.8,2.7Hz,3H),7.48(dd,J＝8.8,2.3Hz,1H),7.37(s,1H),4.33-4.13(m,2H),3.03(s,1H),1.94(d,J＝12.8Hz,2H),1.67(qd,J＝12.2,11.5,3.6Hz,2H),1.25(q,J＝5.5,4.0Hz,2H)；MS(ESI)m/z:calculated for C23H18Cl2N4O4[M+H]+:485.07,found:485.4.。
实施例26
N-((1-(5-氯苯并呋喃-2-羰基)哌啶-4-基)甲基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(I-17)的合成
步骤1：((1-(5-氯苯并呋喃-2-羰基)哌啶-4-基)甲基)氨基甲酸叔丁酯的合成
参照实施例25中步骤1所述的方法，得到淡黄色固体176mg，产率41.92％。1H NMR(300MHz,Chloroform-d)δ(ppm):7.66-7.61(m,1H),7.47(dt,J＝8.8,0.7Hz,1H),7.36(dd,J＝8.8,2.1Hz,1H),7.20(d,J＝0.9Hz,1H),6.83(d,J＝8.2Hz,1H),4.69(s,2H),3.09(s,3H),1.85(d,J＝12.4Hz,3H),1.47(s,9H),1.37-1.26(m,2H).
步骤2：(4-(氨基甲基)哌啶-1-基)(5-氯苯并呋喃-2-基)甲酮的合成
参照实施例2所述的方法，通过硅胶柱层析(二氯甲烷/甲醇＝75/1)纯化得到透明油状液体107mg，产率81.59％。1H NMR(300MHz,Chloroform-d)δ(ppm):7.63(dd,J＝2.2,0.6Hz,1H),7.47(dt,J＝8.8,0.8Hz,1H),7.36(dd,J＝8.8,2.1Hz,1H),7.20(d,J＝0.9Hz,1H),4.58(d,J＝70.8Hz,2H),3.01(d,J＝93.7Hz,2H),2.68(d,J＝6.5Hz,2H),1.90(d,J＝12.8Hz,2H),1.76(s,3H),1.29(dt,J＝9.6,2.6Hz,2H).
步骤3：N-((1-(5-氯苯并呋喃-2-羰基)哌啶-4-基)甲基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(I-17)的合成
参照实施例18中步骤2所述的方法，通过硅胶柱层析(石油醚/乙酸乙酯＝20/1)纯化得到白色固体34mg，产率18.63％。1H NMR(400MHz,Chloroform-d)δ(ppm):8.17-8.08(m,2H),7.64(d,J＝2.1Hz,1H),7.60-7.52(m,2H),7.47(d,J＝8.8Hz,1H),7.39-7.31(m,2H),7.23(d,J＝0.9Hz,1H),4.62(d,J＝79.4Hz,2H),3.49(s,2H),2.97(d,J＝67.1Hz,2H),2.09-2.01(m,1H),1.95(d,J＝13.4Hz,2H),1.47-1.39(m,2H)；MS(ESI)m/z:calculated for C24H20Cl2N4O4[M+H]+:499.09,found:499.2.。
实施例27
5-氯-N-(1-(5-(4-氯苯基)-1,3,4-噁二唑-2-羰基)哌啶-4-基)苯并呋喃-2-甲酰胺(I-18)的合成
步骤1：4-(5-氯苯并呋喃-2-甲酰胺基)哌啶-1-羧酸叔丁酯的合成
参照实施例25中步骤1所述的方法，得到黄色固体280mg，产率72.65％.1H NMR(300MHz,Chloroform-d)δ(ppm):7.67(dd,J＝2.1,0.6Hz,1H),7.48-7.42(m,2H),7.39(dd,J＝8.7,2.1Hz,1H),6.52(d,J＝8.2Hz,1H),4.17(dddd,J＝15.6,11.8,7.8,4.3Hz,3H),3.02-2.90(m,2H),2.05(dd,J＝12.9,3.7Hz,2H),1.71(d,J＝6.7Hz,2H),1.47(s,9H).
步骤2：5-氯-N-(哌啶-4-基)苯并呋喃-2-甲酰胺的合成
参照实施例2所述的方法，通过硅胶柱层析(二氯甲烷/甲醇＝50/1)纯化得到白色固体162mg，产率78.64％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.67(d,J＝8.0Hz,1H),7.87(d,J＝2.2Hz,1H),7.70(d,J＝8.9Hz,1H),7.54(d,J＝0.9Hz,1H),7.48(dd,J＝8.8,2.2Hz,1H),3.85(dtd,J＝11.6,7.5,4.0Hz,1H),3.05-2.93(m,2H),2.57(dd,J＝12.3,2.5Hz,2H),1.79-1.69(m,2H),1.48(qd,J＝12.1,4.1Hz,2H),1.24(dd,J＝6.9,2.6Hz,1H).
步骤3：5-氯-N-(1-(5-(4-氯苯基)-1,3,4-噁二唑-2-羰基)哌啶-4-基)苯并呋喃-2-甲酰胺(I-18)的合成
参照实施例18中步骤2所述的方法，通过硅胶柱层析(石油醚/乙酸乙酯＝1/1)纯化得到淡黄色固体154mg，产率54.60％。1H NMR(400MHz,DMSO-d6)δ(ppm):8.80(d,J＝8.0Hz,1H),8.11-8.05(m,2H),7.88(d,J＝2.2Hz,1H),7.75-7.68(m,3H),7.56(d,J＝0.9Hz,1H),7.49(dd,J＝8.9,2.3Hz,1H),4.52(dd,J＝27.4,13.6Hz,2H),4.20(dtt,J＝11.3,8.0,4.0Hz,1H),3.49-3.41(m,1H),3.16-3.08(m,1H),1.97(td,J＝16.5,15.8,3.9Hz,2H),1.73-1.56(m,2H)；MS(ESI)m/z:calculated for C23H18Cl2N4O4[M+H]+:485.07,found:485.2.。
实施例28
5-氯-N-((1-(5-(4-氯苯基)-1,3,4-噁二唑-2-羰基)哌啶-4-基)甲基)苯并呋喃-2-甲酰胺(I-19)的合成
步骤1：4-((5-氯苯并呋喃-2-甲酰胺基)甲基)哌啶-1-羧酸叔丁酯的合成
参照实施例25中步骤1所述的方法，得到淡黄色固体226mg，产率61.64％。1H NMR (400MHz,Chloroform-d)δ(ppm):7.69-7.66(m,1H),7.46-7.43(m,2H),7.39(dd,J＝8.7,2.1Hz,1H),6.75(d,J＝6.7Hz,1H),4.16(s,2H),3.41(s,2H),2.71(d,J＝13.4Hz,2H),1.85(ddd,J＝10.4,7.0,3.5Hz,1H),1.78(d,J＝13.8Hz,2H),1.47(s,9H),1.26(ddd,J＝16.3,10.0,4.2Hz,2H).
步骤2：5-氯-N-(哌啶-4-基甲基)苯并呋喃-2-甲酰胺的合成
参照实施例2所述的方法，得到白色固体66mg，产率39.19％。1H NMR(300MHz,Chloroform-d)δ(ppm):7.67(d,J＝1.9Hz,1H),7.47-7.41(m,2H),7.39(dd,J＝8.8,2.1Hz,1H),6.74(s,1H),3.40(t,J＝6.2Hz,2H),3.15(dt,J＝12.3,3.4Hz,2H),2.64(td,J＝12.2,2.3Hz,2H),2.16(s,2H),1.79(dt,J＝12.3,3.5Hz,3H),1.33-1.28(m,1H).
步骤3：5-氯-N-((1-(5-(4-氯苯基)-1,3,4-噁二唑-2-羰基)哌啶-4-基)甲基)苯并呋喃-2-甲酰胺(I-19)的合成
参照实施例18中步骤2所述的方法，通过硅胶柱层析(石油醚/乙酸乙酯＝1/1)纯化得到白色固体20mg，产率17.77％。1H NMR(400MHz,Chloroform-d)δ(ppm):8.13-8.08(m,2H),7.68(d,J＝2.0Hz,1H),7.57-7.51(m,2H),7.44(s,2H),7.40(dd,J＝8.9,2.1Hz,1H),6.80(t,J＝6.3Hz,1H),5.03(ddt,J＝13.7,4.5,2.4Hz,1H),4.81(ddt,J＝13.5,4.6,2.4Hz,1H),3.46(t,J＝6.6Hz,2H),3.36-3.23(m,1H),2.91(td,J＝12.9,2.9Hz,1H),2.09(dqd,J＝8.1,6.0,5.1,3.4Hz,1H),2.03-1.96(m,2H),1.44(tdd,J＝13.1,8.7,4.9Hz,2H)；MS(ESI)m/z:calculated for C24H20Cl2N4O4[M+H]+:499.09,found:499.2.。
实施例29
中间体反-(4-(6-氯喹啉-2-甲酰胺基)环己基)氨基甲酸叔丁酯的合成
步骤1：6-氯喹啉-2-甲醛的合成
将2-甲基-6-氯喹啉(200mg，1.13mmol)溶于1,4-二氧六环(5mL)，加入二氧化锡(750mg，6.76mmol)，升温至80℃反应3h。用硅藻土抽滤，滤液减压浓缩，干燥得到粉色固体198mg，产率91.78％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.12(d,J＝0.8Hz,1H),8.59(d,J＝8.5Hz,1H),8.31(d,J＝2.4Hz,1H),8.26(d,J＝9.0Hz,1H),8.05(d,J＝8.5Hz,1H),7.94(dd,J＝9.0,2.4Hz,1H).
步骤2：6-氯喹啉-2-羧酸的合成
将6-氯喹啉-2-甲醛(107mg，0.56mmol)溶于叔丁醇(10mL)，再缓慢滴加溶解有亚氯酸钠(152mg，1.68mmol)和磷酸二氢钠(201mg，1.68mmol)的水溶液(5mL),反应10h。将亚硫酸钠(228mg，2.23mmol)溶于5mL水，接着缓慢加入反应液中淬灭反应。冷却至室温，缓慢滴加1mol/L盐酸，调节pH至4～5，抽滤，干燥，得白色固体73mg，产率62.97％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.54(d,J＝8.6Hz,1H),8.26(d,J＝2.4Hz,1H),8.17(dd,J＝8.8,3.8Hz,2H),7.88(dd,J＝9.0,2.4Hz,1H).
步骤3：反-(4-(6-氯喹啉-2-甲酰胺基)环己基)氨基甲酸叔丁酯的合成
参照实施例1中步骤3所述的方法，得到白色固体102mg，产率71.82％。1H NMR(400MHz,DMSO-d6)δ(ppm):8.68(d,J＝8.5Hz,1H),8.54(d,J＝8.5Hz,1H),8.25(d,J＝2.4Hz,1H),8.18(dd,J＝8.7,2.6Hz,2H),7.89(dd,J＝9.0,2.4Hz,1H),6.79(d,J＝8.0Hz,1H),3.83-3.71(m, 1H),3.25-3.20(m,1H),1.85(d,J＝11.3Hz,4H),1.55(q,J＝12.5Hz,2H),1.39(s,9H),1.31-1.23(m,2H).。
实施例30
反-5-(4-氯苯基)-N-(4-(6-氯喹啉-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-20)的合成
步骤1：反-N-(4-氨基环己基)-6-氯喹啉-2-甲酰胺的合成
参照实施例2所述的方法,得到淡黄色固体167mg，产率73.52％。1H NMR(400MHz,DMSO-d6)δ(ppm):8.71(d,J＝8.4Hz,1H),8.55(d,J＝8.6Hz,1H),8.26(d,J＝2.4Hz,1H),8.17(t,J＝8.9Hz,2H),7.89(dd,J＝9.0,2.5Hz,1H),3.82(dtd,J＝12.0,8.2,4.3Hz,1H),3.08-2.95(m,1H),2.04-1.87(m,4H),1.64-1.39(m,4H).
步骤5：反-5-(4-氯苯基)-N-(4-(6-氯喹啉-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-20)的合成
参照实施例1中步骤3所述的方法,得到白色固体96mg，产率84.03％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.29(d,J＝8.2Hz,1H),8.71(d,J＝8.5Hz,1H),8.55(d,J＝8.6Hz,1H),8.26(d,J＝2.4Hz,1H),8.20(dd,J＝8.8,4.6Hz,2H),8.15-8.08(m,2H),7.89(dd,J＝9.0,2.4Hz,1H),7.76-7.70(m,2H),3.86(d,J＝10.0Hz,2H),1.98-1.89(m,4H),1.69-1.59(m,4H)；MS(ESI)m/z:calculated for C25H21Cl2N5O3[M+H]+:510.10,found:510.5.。
实施例31
反-5-(4-氯苄基)-N-(4-(6-氯喹啉-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-21)的合成
参照实施例1中步骤3所述的方法,得到白色固体43mg，产率49.82％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.21(d,J＝8.3Hz,1H),8.69(d,J＝8.5Hz,1H),8.55(d,J＝8.6Hz,1H),8.25(d,J＝2.5Hz,1H),8.19(dd,J＝8.8,4.9Hz,2H),7.88(dd,J＝9.0,2.5Hz,1H),7.47-7.42(m,2H),7.41-7.37(m,2H),4.39(s,2H),3.81(s,2H),1.94-1.84(m,4H),1.60(q,J＝11.9,11.1Hz,4H)；MS(ESI)m/z:calculated for C26H23Cl2N5O3[M+H]+:524.12,found:524.5.。
实施例32
反-5-(4-溴苯基)-N-(4-(6-氯喹啉-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-22)的合成
参照实施例1中步骤3所述的方法,得到白色固体23mg，产率21.34％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.74(d,J＝8.6Hz,1H),8.55(d,J＝8.8Hz,1H),8.40(d,J＝7.9Hz,1H),8.29-8.14(m,3H),7.90(d,J＝9.1Hz,1H),7.81(d,J＝8.5Hz,2H),7.68(d,J＝8.0Hz,2H),3.84(s,2H),1.92(d,J＝11.5Hz,4H),1.65(d,J＝12.5Hz,2H),1.49(d,J＝12.3Hz,2H)；MS(ESI)m/z:calculated for C25H21BrClN5O3[M+H]+:554.05,found:554.1.。
实施例33
反-5-(4-氯-3-氟苯基)-N-(4-(6-氯喹啉-2-甲酰胺基)环己基)-1,3,4-噁二唑-2-甲酰胺(Ⅰ-23)的合成
参照实施例1中步骤3所述的方法,得到白色固体47mg，产率54.05％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.30(d,J＝8.2Hz,1H),8.71(d,J＝8.4Hz,1H),8.55(d,J＝8.5Hz,1H),8.25(d,J＝2.4Hz,1H),8.20(dd,J＝8.8,4.7Hz,2H),8.12(dd,J＝9.5,1.9Hz,1H),7.97(dd,J＝8.4,1.9Hz,1H),7.93-7.87(m,2H),3.94-3.81(m,2H),1.94(d,J＝8.3Hz,4H),1.70-1.59(m,4H)；MS(ESI)m/z:calculated for C25H20Cl2FN5O3[M+H]+:528.09,found:528.4.。
实施例34
反-3-(4-氯苯基)-N-(4-(6-氯喹啉-2-甲酰胺基)环己基)-1,2,4-噁二唑-5-甲酰胺(Ⅰ-24)的合成
参照实施例1中步骤3所述的方法,得到白色固体57mg，产率53.85％。1H NMR(400MHz,DMSO-d6)δ(ppm):9.43(d,J＝8.2Hz,1H),8.71(d,J＝8.5Hz,1H),8.55(d,J＝8.6Hz,1H),8.26(d,J＝2.4Hz,1H),8.20(dd,J＝8.8,4.6Hz,2H),8.12-8.07(m,2H),7.89(dd,J＝9.0,2.4Hz,1H),7.73-7.69(m,2H),3.86(s,2H),1.94(d,J＝7.9Hz,4H),1.65(q,J＝9.9,8.4Hz,4H)；MS(ESI)m/z:calculated for C25H21Cl2N5O3[M+H]+:510.10,found:510.4.。
实施例35
反-6-氯-N-(4-(((5-(4-氯苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)喹啉-2-甲酰胺(Ⅰ-25)的合成
参照实施例13中步骤3所述的方法,得到白色固体34mg，产率49.36％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.68(d,J＝8.5Hz,1H),8.54(d,J＝8.7Hz,1H),8.25(d,J＝2.4Hz,1H),8.18(dd,J＝8.8,3.0Hz,2H),8.05-7.99(m,2H),7.88(dd,J＝9.1,2.4Hz,1H),7.74-7.69(m,2H),5.33(s,2H),3.80(d,J＝10.4Hz,1H),2.00(d,J＝6.8Hz,1H),1.88(d,J＝11.8Hz,4H),1.64-1.51(m,2H),1.36(t,J＝11.9Hz,2H).；MS(ESI)m/z:calculated for C25H23Cl2N5O2[M+H]+:496.12,found:496.2.。
实施例36
反-6-氯-N-(4-(((5-(4-氯苄基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)喹啉-2-甲酰胺(Ⅰ-26)的合成
参照实施例13中步骤3所述的方法,得到白色固体88mg，产率65.47％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.62(d,J＝8.5Hz,1H),8.54(d,J＝8.6Hz,1H),8.25(d,J＝2.4Hz,1H),8.17(dd,J＝8.8,6.2Hz,2H),7.88(dd,J＝9.1,2.4Hz,1H),7.46-7.42(m,2H),7.38-7.33(m,2H),4.29(s,2H),3.92(s,2H),3.80(d,J＝11.5Hz,1H),2.41-2.32(m,1H),1.92-1.80(m,4H),1.51-1.40(m,2H),1.12(dd,J＝14.8,4.5Hz,2H)；MS(ESI)m/z:calculated for C26H25Cl2N5O2[M+H]+:510.14,found:510.5.。
实施例37
反-N-(4-(((5-(4-溴苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)-6-氯喹啉-2-甲酰胺(Ⅰ-27)的合成
参照实施例13中步骤3所述的方法,得到白色固体73mg，产率51.25％。1H NMR(400MHz,DMSO-d6)δ(ppm):8.65(d,J＝8.5Hz,1H),8.54(d,J＝8.5Hz,1H),8.24(d,J＝2.4Hz,1H),8.18(dd,J＝8.8,5.8Hz,2H),7.96-7.92(m,2H),7.88(dd,J＝9.1,2.4Hz,1H),7.86-7.82(m,2H),5.33(s,2H),3.83-3.76(m,1H),2.07-1.93(m,1H),1.89(d,J＝11.7Hz,4H),1.62-1.53(m,2H),1.38-1.33(m,2H).；MS(ESI)m/z:calculated for C25H23BrClN5O2[M+H]+:540.07,found:540.8.。
实施例38
反-6-氯-N-(4-(((5-(4-氯-3-氟苯基)-1,3,4-噁二唑-2-基)甲基)氨基)环己基)喹啉-2-甲酰胺(Ⅰ-28)的合成
参照实施例13中步骤3所述的方法,得到白色固体22mg，产率21.66％。1H NMR(400MHz,DMSO-d6)δ(ppm):8.68(d,J＝8.5Hz,1H),8.54(d,J＝8.6Hz,1H),8.25(d,J＝2.4Hz,1H), 8.18(dd,J＝8.8,4.1Hz,2H),8.04-7.99(m,1H),7.92-7.86(m,3H),5.34(s,2H),3.80(d,J＝9.0Hz,1H),2.06-1.95(m,1H),1.88(d,J＝11.5Hz,4H),1.58(q,J＝11.8Hz,2H),1.36(t,J＝11.2Hz,2H)；MS(ESI)m/z:calculated for C25H22Cl2FN5O2[M+H]+:514.11,found:514.0.。
实施例39
中间体2-(5-氯苯并呋喃-2-基)-5-(哌啶-4-基)-1,3,4-噁二唑的合成
步骤1：5-氯苯并呋喃-2-碳酰肼的合成
参照实施例3中步骤1所述的方法，得到白色固体248mg，产率95.15％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.13(s,1H),7.87(d,J＝2.2Hz,1H),7.69(d,J＝8.8Hz,1H),7.52-7.43(m,2H),4.63(s,2H).
步骤2：4-(2-(5-氯苯并呋喃-2-羰基)肼-1-羰基)哌啶-1-羧酸叔丁酯的合成
参照实施例1中步骤3所述的方法,得到淡黄色固体180mg，产率95.60％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.68(s,1H),10.03(s,1H),7.92(d,J＝2.2Hz,1H),7.74(d,J＝8.8Hz,1H),7.64(d,J＝0.9Hz,1H),7.52(dd,J＝8.9,2.2Hz,1H),3.97(d,J＝13.1Hz,2H),2.77(d,J＝19.6Hz,2H),2.49-2.39(m,1H),1.73(d,J＝12.8Hz,2H),1.54–1.43(m,2H),1.41(s,9H).
步骤3：2-(5-氯苯并呋喃-2-基)-5-(哌啶-4-基)-1,3,4-噁二唑的合成
参照实施例2所述的方法，得到淡黄色固体102mg，产率78.71％。MS(ESI)m/z:calculated for C15H14ClN3O2[M+H]+:304.08,found:304.43.。
实施例39
1-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)哌啶-1-基)-2-(4-氯苯氧基)乙烷-1-酮(II-1)的合成
参照实施例1中步骤3所述的方法,纯化得到白色固体44mg，产率27.74％。1H NMR(300MHz,DMSO-d6)δ(ppm):7.91(d,J＝2.2Hz,1H),7.85(d,J＝8.9Hz,1H),7.78(s,1H),7.55(dd,J＝8.9,2.2Hz,1H),7.37–7.30(m,2H),7.00–6.93(m,2H),4.90(d,J＝4.0Hz,2H),4.26(d,J＝13.3Hz,1H),3.88(d,J＝13.8Hz,1H),3.46(d,J＝6.7Hz,1H),3.27(d,J＝12.2Hz,1H),2.97(t,J＝12.2Hz,1H),2.13(s,2H),1.87(q,J＝10.8Hz,1H),1.67(q,J＝10.0Hz,1H).；MS(ESI)m/z:calculated for C23H19Cl2N3O4[M+H]+:472.08,found:472.4.。
实施例40
反-N-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己基)-2-(4-氯苯氧基)乙酰胺(II-2)的合成
步骤1：反-4-(2-(5-氯苯并呋喃-2-羰基)肼-1-羰基)环己基)氨基甲酸叔丁酯的合成
参照实施例1中步骤3所述的方法,得到淡黄色固体160mg，产率80.53％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.65(s,1H),9.92(s,1H),7.92(d,J＝2.2Hz,1H),7.74(d,J＝8.9Hz,1H),7.63(d,J＝0.9Hz,1H),7.51(dd,J＝8.9,2.2Hz,1H),6.77(d,J＝8.0Hz,1H),3.18(s,1H),2.16(t,J＝12.1Hz,1H),1.81(t,J＝12.4Hz,4H),1.38(d,J＝2.7Hz,9H),1.21(dq,J＝24.2,12.2Hz,4H).
步骤2：反-4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己-1-胺的合成
参照实施例2所述的方法，得到淡黄色固体99mg，产率72.62％。MS(ESI)m/z:calculated for C15H14ClN3O2[M+H]+:318.09,found:318.06
步骤3：反-N-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己基)-2-(4-氯苯氧基)乙酰胺(Ⅲ-2)的合成
参照实施例1中步骤3所述的方法,得到乳白色固体57mg，产率37.62％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.07(d,J＝8.2Hz,1H),7.90(s,1H),7.84(d,J＝8.9Hz,1H),7.77(d,J＝6.9Hz,1H),7.54(d,J＝9.0Hz,1H),7.40-7.29(m,2H),6.98(dt,J＝14.0,6.7Hz,2H),4.49(s,2H),3.72(d,J＝9.2Hz,1H),3.11-2.93(m,1H),2.19(d,J＝13.5Hz,2H),1.91(d,J＝12.3Hz,2H),1.67(d,J＝12.4Hz,2H),1.47(q,J＝12.6Hz,2H)；MS(ESI)m/z:calculated for C24H21Cl2N3O4[M+H]+:486.09,found:486.4.。
实施例41
反-N-((4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己基)甲基)-2-(4-氯苯氧基)乙酰胺(II-3)的合成
步骤1：反-((4-(2-(5-氯苯并呋喃-2-羰基)肼-1-羰基)环己基)甲基)氨基甲酸叔丁酯的合成
参照实施例1中步骤3所述的方法,得到淡黄色固体364mg，产率85.20％。1H NMR(300MHz,DMSO-d6)δ(ppm):10.63(s,1H),9.91(s,1H),7.92(d,J＝2.2Hz,1H),7.74(d,J＝8.9Hz,1H),7.63(d,J＝0.9Hz,1H),7.51(dd,J＝8.9,2.3Hz,1H),6.83(t,J＝5.9Hz,1H),2.92-2.67(m,3H),2.19(t,J＝12.0Hz,1H),1.86-1.67(m,4H),1.38(s,9H),1.32(d,J＝10.5Hz,2H),0.98-0.83(m,2H).
步骤2：反-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己基)甲胺的合成
参照实施例2所述的方法，得到淡黄色固体235mg，产率88.77％。MS(ESI)m/z:calculated for C15H14ClN3O2[M+H]+:332.11,found:332.92.
步骤3：反-N-((4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己基)甲基)-2-(4-氯苯氧基)乙酰胺(II-3)的合成
参照实施例1中步骤3所述的方法,得到白色固体108mg，产率30.47％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.16(t,J＝6.0Hz,1H),7.90(d,J＝2.2Hz,1H),7.84(d,J＝8.9Hz,1H),7.76(d,J＝1.0Hz,1H),7.54(dd,J＝8.9,2.2Hz,1H),7.39-7.33(m,2H),7.03-6.97(m,2H),4.52(s,2H),3.04(t,J＝6.4Hz,2H),3.01-2.92(m,1H),2.15(d,J＝11.7Hz,2H),1.79(d,J＝12.7Hz,2H),1.54(dd,J＝13.5,10.1Hz,3H),1.07(d,J＝12.3Hz,2H)；MS(ESI)m/z:calculated for C25H23Cl2N3O4[M+H]+:500.11,found:500.4.。
实施例42
反-N-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己基)-5-(4-氯苯基)-1,3,4-噁二唑-2-甲酰胺(II-4)的合成
参照实施例1中步骤3所述的方法,得到白色固体33mg，产率35.06％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.45(t,J＝6.1Hz,1H),8.15-8.08(m,2H),7.90(d,J＝2.2Hz,1H),7.84(d,J＝8.8Hz,1H),7.75(d,J＝5.0Hz,2H),7.73-7.70(m,1H),7.54(dd,J＝8.9,2.2Hz,1H),3.21(d,J＝6.5Hz,2H),3.04(s,1H),2.19(d,J＝12.7Hz,2H),1.90(d,J＝13.0Hz,2H),1.66(d,J＝18.3Hz,1H),1.52(d,J＝12.9Hz,2H)；MS(ESI)m/z:calculated for C26H21Cl2N5O4[M+H]+:538.10,found:538.5.。
实施例43
反-N-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-噁二唑-2-基)环己基)-3-(4-氯苯基)-1,2,4-噁二唑-5-甲酰胺(II-5)的合成
参照实施例1中步骤3所述的方法,得到白色固体30mg，产率23.11％。1H NMR(300MHz,DMSO-d6)δ(ppm):9.56(t,J＝6.1Hz,1H),8.13-8.06(m,2H),7.90(d,J＝2.2Hz,1H),7.84(d,J＝8.9Hz,1H),7.76(d,J＝0.9Hz,1H),7.74-7.65(m,2H),7.54(dd,J＝8.9,2.2Hz,1H),3.22(t,J＝6.5Hz,2H),3.04(s,1H),2.19(d,J＝12.7Hz,2H),1.90(d,J＝13.0Hz,2H),1.70(s,1H),1.55(t,J＝12.9Hz,2H),1.21-1.13(m,2H)；MS(ESI)m/z:calculated for C26H21Cl2N5O4[M+H]+:538.10,found:538.4.。
实施例44
中间体4-(5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)环己烷-1-胺的合成
步骤1：5-氯-1H-吲哚-2-碳酰肼的合成
参照实施例39步骤1中所述的方法，得黄白色固体308mg，产率：91.33％。1H NMR(400MHz,DMSO-d6)δ11.90–11.77(m,1H),9.88(s,1H),7.67(d,J＝2.0Hz,1H),7.43(d,J＝ 8.7Hz,1H),7.17(dd,J＝8.7,2.1Hz,1H),7.07(d,J＝2.0Hz,1H).
步骤2：(4-(2-(5-氯-1H-吲哚-2-羰基)肼-1-羰基)环己基)氨基甲酸叔丁酯的合成
参照实施例39步骤2中所述的方法，得白色固体408mg，产率99.74％。1H NMR(300MHz,DMSO-d6)δ11.89(d,J＝2.1Hz,1H),10.42(s,1H),9.86(s,1H),7.73(d,J＝2.1Hz,1H),7.45(d,J＝8.7Hz,1H),7.25–7.15(m,2H),6.77(d,J＝8.1Hz,1H),3.10(m,1H)2.17(t,J＝11.9Hz,1H),1.80(d,J＝12.2Hz,4H),1.38(s,9H),1.38(m,2H),1.20(m,2H).
步骤3：4-(5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)环己烷-1-胺的合成
参照实施例39步骤3中所述的方法，得黄白色固体205mg，产率86％。1H NMR(500MHz,Chloroform-d)δ9.61(s,1H),7.75(s,1H),7.63(s,1H),7.38(s,1H),7.06(s,1H),5.69(s,1H),3.81(s,1H),3.24(s,1H),2.96(s,1H),1.80–1.69(m,4H),1.70–1.60(m,4H).。
实施例45
N-(4-(5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)环己基)-2-(4-氯苯氧基)乙酰胺(Ⅱ-6)的合成
参照实施例1中步骤3所述的方法,得到白色固体23mg，产率22.53％。1H NMR(300MHz,DMSO-d6)δ12.46(s,1H),8.07(d,J＝8.0Hz,1H),7.74(d,J＝2.0Hz,1H),7.48(d,J＝8.8Hz,1H),7.37(d,J＝2.3Hz,1H),7.35(d,J＝2.3Hz,1H),7.26(dd,J＝8.7,2.1Hz,1H),7.15(s,1H),7.00(d,J＝2.3Hz,1H),6.98(d,J＝2.3Hz,1H),4.49(s,2H),3.71(ddd,J＝11.6,7.6,3.8Hz,1H),3.00(tt,J＝11.6,3.5Hz,1H),2.20(d,J＝11.7Hz,2H),1.91(dd,J＝12.7,3.8Hz,2H),1.67(qd,J＝13.0,3.1Hz,2H),1.57–1.37(m,2H).MS(ESI)m/z:calculated for C24H22Cl2N4O3[M+H]+:485.11,found:485.4.。
实施例46
(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)(5-(4-氯苯基)-1,3,4恶二唑-1-基)甲酮(Ⅱ-7)的合成
参照实施例1中步骤3所述的方法,得到白色固体54mg，产率40.00％。1H NMR(300MHz,DMSO-d6)δ8.10(d,J＝2.0Hz,1H),8.07(d,J＝2.0Hz,1H),7.91(d,J＝2.2Hz,1H),7.85(d,J＝8.9Hz,1H),7.79(d,J＝0.9Hz,1H),7.74(d,J＝2.0Hz,1H),7.72(d,J＝2.0Hz,1H),7.55(dd,J＝8.9,2.2Hz,1H),4.50(dd,J＝43.3,13.4Hz,2H),3.69–3.49(m,2H),3.32–3.25(m,1H),2.31–2.16(m,2H),2.05–1.79(m,2H).MS(ESI)m/z:calculated for C24H17Cl2N5O4[M+H]+510.07,found:510.33.。
实施例47
中间体2-(5-氯-1H-吲哚-2-基)-5-(哌啶-4-基)-1,3,4-恶二唑的合成
步骤1：4-(2-(5-氯-1H-吲哚-2-羰基)肼-1-羰基)哌啶-1-甲酸叔丁酯的合成
原料为1-(叔丁氧羰基)哌啶-4-羧酸，参照实施例1中步骤3的合成得黄白色固体473mg， 产率为99.72％。1H NMR(300MHz,DMSO-d6)δ11.90(d,J＝2.1Hz,1H),10.46(s,1H),9.97(s,1H),7.74(d,J＝2.0Hz,1H),7.45(d,J＝8.7Hz,1H),7.26–7.16(m,2H),3.97(d,J＝13.1Hz,2H),2.71(s,1H),1.81–1.69(m,2H),1.50(td,J＝12.3,4.1Hz,2H),1.41(s,9H),1.23(t,J＝8.2Hz,2H).
步骤2：2-(5-氯-1H-吲哚-2-基)-5-(哌啶-4-基)-1,3,4-恶二唑的合成
参照实施例39中步骤3的合成得棕黄色固体156mg，产率91.92％。1H NMR(400MHz,DMSO-d6)δ11.93(s,1H),7.74(d,J＝2.1Hz,1H),7.45(d,J＝8.7Hz,1H),7.23–7.18(m,2H),3.10(s,4H),2.85(s,1H),2.56(s,1H),1.88(d,J＝17.2Hz,4H).。
实施例48
(4-(5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)(5-(4-氯苯基)-1,3,4恶二唑-1-基)甲酮(Ⅱ-8)的合成
参照实施例1中步骤3所述的方法,得到白色固体53mg，产率36.63％。1H NMR(500MHz,Chloroform-d)δ9.61(s,1H),8.14–8.08(m,3H),7.62(s,1H),7.56(s,1H),7.48–7.39(m,4H),7.17(s,1H),4.50(dd,J＝43.3,13.4Hz,2H),3.69–3.49(m,2H),3.32–3.25(m,1H),2.31–2.16(m,2H),2.05–1.79(m,2H).MS(ESI)m/z:calculated for C24H18Cl2N6O3[M+H]+509.08,found：509.35.。
实施例49
中间体(1r，4r)-4-((5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)甲基)环己烷-1-胺的合成
步骤1：叔丁基((1r，4r)-4-(2-(2-(5-氯-1H-吲哚-2-羰基)肼基)-2-氧乙基)环己基)氨基甲酸酯的合成
原料为实施例44步骤1合成的5-氯-1H-吲哚-2-碳酰肼、(1r，4r)-4-((叔丁氧羰基)氨基)环己烷-1-甲酸。参照实施例1中步骤3的合成，得黄白色固体374mg，产率为78.72％。1H NMR(300MHz,DMSO-d6)δ11.94(s,1H),10.40(s,1H),9.87(s,1H),7.74(d,J＝2.0Hz,1H),7.44(d,J＝8.7Hz,1H),7.29–7.15(m,2H),6.70(d,J＝8.1Hz,1H),3.18(d,J＝10.4Hz,1H),2.07(d,J＝6.9Hz,2H),1.78(s,4H),1.62(m,1H),1.38(s,9H),1.19(d,J＝12.6Hz,1H),1.16–0.92(m,3H).
步骤2：(1r，4r)-4-((5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)甲基)环己烷-1-胺的合成
参照实施例39中步骤3的合成得棕黄色固体48mg，产率30.27％。MS(ESI)m/z:calculated for C17H19ClN4O[M+H]+:330.82,found:330.83。
实施例50
N-((1r，4r)-4-((5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)甲基)环己基)-2-(4-氯苯氧基)乙酰胺(Ⅱ-9)的合成
参照实施例1中步骤3所述的方法,得到白色固体27mg，产率74.32％。1H NMR(300MHz,DMSO-d6)δ12.45(s,1H),7.95(d,J＝8.1Hz,1H),7.74(d,J＝2.0Hz,1H),7.49(d,J＝8.7Hz,1H),7.40–7.28(m,2H),7.26(dd,J＝8.8,2.1Hz,1H),7.15(s,1H),7.03–6.91(m,2H),4.45(s,2H),3.61(d,J＝8.3Hz,1H),2.87(d,J＝6.4Hz,2H),1.80(d,J＝8.3Hz,5H),1.40–1.23(m,2H),1.28–1.08(m,2H).MS(ESI)m/z:calculated for C25H24Cl2N4O3[M+H]+:499.12,found:499.39。
实施例51
中间体(1r，4r)-4-((5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)甲基)环己烷-1-胺的合成
步骤1：叔丁基((1r，4r)-4-(2-(2-(5-氯苯并呋喃-2-羰基)肼基)-2-氧乙基)环己基)氨基甲酸酯的合成
原料为实施例39步骤1合成的5-氯苯并呋喃-2-碳酰肼、(1r，4r)-4-((叔丁氧羰基)氨基)环己烷-1-甲酸。参照实施例1中步骤3的合成，得白色固体582mg，产率为82.21％。1H NMR(300MHz,DMSO-d6)δ10.40(s,1H),9.87(s,1H),7.74(d,J＝2.0Hz,1H),7.44(d,J＝8.7Hz,1H),7.29–7.15(m,2H),6.70(d,J＝8.1Hz,1H),3.18(d,J＝10.4Hz,1H),2.07(d,J＝6.9Hz,2H),1.78(s,4H),1.62(m,1H),1.38(s,9H),1.19(d,J＝12.6Hz,1H),1.16–0.92(m,3H).
步骤2：(1r，4r)-4-((5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)甲基)环己烷-1-胺的合成。
参照实施例39中步骤3的合成得棕黄色固体92mg，产率73％。1H NMR(300MHz,DMSO-d6)δ7.93–7.80(m,2H),7.76(s,1H),7.54(dd,J＝8.8,2.2Hz,1H),2.87(d,J＝6.4Hz,2H),1.75(s,5H),1.10(d,J＝18.2Hz,5H).。
实施例52
N-((1r，4r)-4-((5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)甲基)环己基)-2-(4-氯苯氧基)乙酰胺(Ⅱ-10)的合成
参照实施例1中步骤3所述的方法,得到白色固体53mg，产率84.32％。1H NMR(400MHz,DMSO-d6)δ7.93(d,J＝8.1Hz,1H),7.89(d,J＝2.2Hz,1H),7.84(d,J＝8.8Hz,1H),7.75(d,J＝0.9Hz,1H),7.54(dd,J＝8.9,2.2Hz,1H),7.41–7.29(m,2H),7.04–6.92(m,2H),3.61(dtt,J＝11.5,7.8,3.9Hz,1H),2.90(d,J＝6.5Hz,2H),1.78(td,J＝9.6,8.7,4.4Hz,5H),1.38–1.09(m,4H).MS(ESI)m/z:calculated for C25H23Cl2N3O4[M+H]+:500.11,found:500.39。
实施例53
中间体2-(4-氯苯氧基)-N-((1r，4r)-4-(肼羰基)环己基)乙酰胺的合成
步骤1：(1r，4r)-4-(2-(4-氯苯氧基)乙酰胺基)环己烷-1-甲酸甲酯的合成
原料为(1r，4r)-4-氨基环己烷-1-甲酸甲酯，参照实施例1中步骤3所述的方法。得白色固体335mg，产率72.24％。1H NMR(300MHz,DMSO-d6)δ7.97(d,J＝7.8Hz,1H),7.40–7.29(m,2H),7.03–6.92(m,2H),4.45(s,2H),3.59(s,3H),2.25(ddt,J＝11.6,7.1,3.5Hz,1H),2.00–1.85(m,2H),1.85–1.71(m,2H),1.49–1.13(m,5H).
步骤2：2-(4-氯苯氧基)-N-((1r，4r)-4-(肼羰基)环己基)乙酰胺的合成
参照实施例1中步骤3所述的方法,得到白色固体273mg，产率72.47％。1H NMR(300MHz,DMSO-d6)δ8.93(s,1H),7.95(d,J＝8.1Hz,1H),7.44–7.26(m,2H),7.05–6.89(m,2H),4.45(s,2H),4.14(s,2H),3.68–3.48(m,1H),2.00(tt,J＝12.0,3.5Hz,1H),1.86–1.60(m,4H),1.54–1.34(m,2H),1.34–1.13(m,2H).。
实施例54
2-(4-氯苯氧基)-N-((1r，4r)-4-(5-(6-氯喹啉-2-基)-1,3,4-恶二唑-2-基)环己基)乙酰胺(Ⅱ-11)的合成
步骤1：2-(4-氯苯氧基)-N-((1r，4r)-4-(2-(6-氯喹啉-2-羰基)肼-1-羰基)环己基)乙酰胺的合成
原料为实施例53步骤2的2-(4-氯苯氧基)-N-((1r，4r)-4-(肼羰基)环己基)乙酰胺、实施例29步骤2的6-氯喹啉-2-羧酸。参照实施例1中步骤3所述的方法。得白色固体261mg，产率72％。1H NMR(300MHz,DMSO-d6)δ10.60(s,1H),10.06(s,1H),8.58(d,J＝8.6Hz,1H),8.28(d,J＝2.4Hz,1H),8.22–8.09(m,2H),8.06–7.86(m,2H),7.43–7.25(m,2H),7.07–6.88(m,2H),4.47(s,2H),3.72–3.53(m,1H),2.26(t,J＝11.3Hz,1H),1.85(d,J＝11.2Hz,4H),1.59–1.18(m,4H).
步骤2：2-(4-氯苯氧基)-N-((1r，4r)-4-(5-(6-氯喹啉-2-基)-1,3,4-恶二唑-2-基)环己基)乙酰胺的合成(Ⅱ-11)
参照实施例39中步骤3的合成得白色固体92mg，产率20.00％。1H NMR(400MHz,DMSO-d6)δ8.61(d,J＝8.7Hz,1H),8.38–8.26(m,2H),8.19(d,J＝9.0Hz,1H),8.05(d,J＝8.0Hz,1H),7.90(dd,J＝9.0,2.4Hz,1H),7.44–7.30(m,2H),7.08–6.93(m,2H),4.49(s,2H),3.74(dtd,J＝11.9,8.0,4.0Hz,1H),3.09(tt,J＝12.0,3.7Hz,1H),2.28–2.17(m,2H),1.93(dd,J＝13.2,3.7Hz,2H),1.71(qd,J＝13.0,3.3Hz,2H),1.49(qd,J＝12.8,3.6Hz,2H).MS(ESI)m/z:calculated for C25H22Cl2N4O3[M+H]+:497.11,found:497.39.。
实施例55
中间体6-氯喹啉-2-羧酰胺的合成
步骤1：6-溴喹啉-2-甲酰胺的合成
将6-溴喹啉-2-羧酸(1.49g,5.90mmol)溶于无水乙腈(20ml)中，在冰浴条件下加入草酰氯(900μl,10.63mmol)，再加入5滴N,N-二甲基甲酰胺，反应3h。将反应混合物加入在冰浴条件下制得的氨水(1.14ml，29.57mmol)：乙腈＝1：10的溶液中，20min后反应完全。抽滤，烘干，得淡黄色固体1.16g，产率72.64％。1H NMR(300MHz,DMSO-d6)δ8.53(d,J＝8.5Hz,1H),8.40(d,J＝2.2Hz,1H),8.33(s,1H),8.19(d,J＝8.6Hz,1H),8.05(d,J＝9.0Hz,1H),7.98(dd,J＝9.0,2.2Hz,1H),7.84(s,1H).
步骤2:6-溴喹啉-2-腈的合成
将6-溴喹啉-2-甲酰胺(500mg，1.99mmol)溶于10mlN,N-二甲基甲酰胺中，随后加入五氯化磷(1.24g，5.97mmol)，于50℃下反应1h。加水抽滤。滤饼用二氯甲烷溶解，将滤液浓缩，残留物通过硅胶柱层析(石油醚/乙酸乙酯＝20/1)纯化得到白色固体，产率33.83％。1H NMR(300MHz,DMSO-d6)δ8.64(d,J＝8.5Hz,1H),8.52–8.43(m,1H),8.16–8.01(m,3H).
步骤3：6-溴喹啉-2-羧酰胺的合成
将6-溴喹啉-2-腈(220mg，943.93μmol)加到10ml无水甲醇中，随后加入甲醇钠(51mg，943.93μmol)，于40℃下反应30min。加入NH4Cl(151.47mg，2.83mmol)，反应3h，NH3·H2O调节PH～8。硅藻土抽滤，用少量甲醇洗涤。将滤液浓缩，残留物用二氯甲烷打浆析出固体，抽滤得白色固体，产率86.51％。1H NMR(300MHz,DMSO-d6)δ8.74(d,J＝8.7Hz,1H),8.64–8.45(m,2H),8.12(d,J＝9.0Hz,1H),8.08(dd,J＝9.1,2.0Hz,1H),7.73(s,1H),6.26(s,2H).。
实施例56
2-(4-氯苯氧基)-N-((1r，4r)-4-(5-(6-溴喹啉-2-基)-4H-1,2,4-三唑-3-基)环己基)乙酰胺(Ⅱ-12)的合成
原料为实施例55步骤3的6-溴喹啉-2-羧酰胺、实施例53步骤2的2-(4-氯苯氧基)-N-((1r，4r)-4-(肼羰基)环己基)乙酰胺。
将6-溴喹啉-2-羧酰胺(105mg，322.29μmol)、2-(4-氯苯氧基)-N-((1r，4r)-4-(肼羰基)环己基)乙酰胺(78mg,322.29μmol)投入封管，用N,N-二甲基甲酰胺溶解，于130℃下反应5h。用乙酸乙酯萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，残留物通过硅胶柱层析纯化得到白色固体17mg，产率9.75％。1H NMR(300MHz,DMSO-d6)δ14.35(d,J＝169.1Hz,1H),8.48(dd,J＝30.8,8.6Hz,1H),8.36(d,J＝20.6Hz,1H),8.29–8.18(m,1H),8.07(d,J＝9.3Hz,1H),8.02(s,1H),7.94(dd,J＝16.1,8.9Hz,1H),7.35(dd,J＝7.7,5.4Hz,2H),7.09–6.91(m,2H),4.47(d,J＝11.2Hz,2H),3.71(s,1H),2.95–2.65(m,1H),2.11(d,J＝12.7Hz,2H),1.90(d,J＝12.0Hz,2H),1.56(dq,J＝61.1,12.9,12.1Hz,4H).MS(ESI)m/z:calculated for C25H23BrClN5O2[M+H]+:541.85,found:542.07.。
实施例57
(5-(4-氯-3-氟苯基)-1,3,4-恶二唑-2-基)(4-(5-(5-氯苯并呋喃-2-基)-1,3,44-恶二唑-1-基)哌啶-1-基)甲酮(Ⅱ-13)的合成
参照实施例1中步骤3所述的方法,得到白色固体72mg，产率87.2％。1H NMR(400MHz,DMSO-d6)δ7.91(d,J＝2.3Hz,1H),7.90–7.87(m,1H),7.85(d,J＝8.8Hz,1H),7.82–7.70(m,3H),7.55(dd,J＝8.8,2.2Hz,1H),4.26(d,J＝13.3Hz,1H),4.09(d,J＝13.6Hz,1H),3.59–3.39(m,2H),3.11(t,J＝11.4Hz,1H),2.27–2.13(m,2H),1.93(t,J＝10.9Hz,1H),1.78(q,J＝10.1Hz,1H).MS(ESI)m/z:calculated for C24H16Cl2FN5O4，[M+H]+:527.06,found:527.35。
实施例58
(4-(5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)(5-(4-氯-3-氟苯基)-1,3,44-恶二唑-1-基)甲酮(Ⅱ-14)的合成
原料为实施例47步骤2的2-(5-氯-1H-吲哚-2-基)-5-(哌啶-4-基)-1,3,4-恶二唑、实施例10步骤1的5-(4-氯-3-氟苯基)-1,3,4-恶二唑-2-羧酸
参照实施例1中步骤3所述的方法,得到白色固体27mg，产率72.17％。1H NMR(400MHz,DMSO-d6)δ12.46(d,J＝2.1Hz,1H),8.08(dd,J＝9.5,1.9Hz,1H),7.98–7.86(m,2H),7.75(d,J＝2.0Hz,1H),7.49(d,J＝8.7Hz,1H),7.27(dd,J＝8.8,2.0Hz,1H),7.22–7.16(m,1H),4.48(dd,J＝40.9,13.6Hz,2H),3.69–3.47(m,2H),2.31–2.15(m,2H),2.04–1.82(m,2H),0.86(t,J＝6.7Hz,1H).MS(ESI)m/z:calculated for C24H17Cl2FN6O3[M+H]+:527.07,found:527.31。
实施例59
中间体5-氯苯呋喃-2-羧酰胺的合成
步骤1：5-氯苯呋喃-2-甲酰胺的合成
参照实施例55步骤1所述的方法，得白色固体2.12g，产率87.24％。1H NMR(300MHz,Chloroform-d)δ9.12(s,2H),7.69(dd,J＝1.9,0.9Hz,1H),7.38(dt,J＝8.8,0.8Hz,1H),7.29(dd,J＝8.8,2.0Hz,1H),7.18(dd,J＝2.1,0.9Hz,1H).
步骤2：5-氯苯呋喃-2-腈的合成
参照实施例55步骤2所述的方法，得白色固体779mg，产率77.41％。1H NMR(300MHz,DMSO-d6)δ8.09(d,J＝1.0Hz,1H),7.96(dd,J＝2.2,0.6Hz,1H),7.81(dt,J＝9.0,0.8Hz,1H),7.64(dd,J＝8.9,2.2Hz,1H).
步骤3：5-氯苯呋喃-2-羧酰胺的合成
参照实施例55步骤3所述的方法，得白色固体257mg，产率98.73％。1H NMR(300MHz,DMSO-d6)δ8.30(s,1H),8.25(s,1H),8.17–8.12(m,2H),8.07(d,J＝2.2Hz,1H),7.78(d,J＝8.9Hz,1H),7.62(dd,J＝8.9,2.2Hz,1H).。
实施例60
N-((1r，4r)-4-(5-(5-氯苯并呋喃-2-基)-4H-1,2,4-三唑-3-基)环己基)-2-(4-氯苯氧基)乙酰胺(Ⅱ-15)的合成
参照实施例56，得白色固体21mg，产率30.20％。1H NMR(300MHz,DMSO-d6)δ14.18(s,1H),8.07(d,J＝8.1Hz,1H),7.78(d,J＝2.2Hz,1H),7.71(d,J＝8.8Hz,1H),7.43–7.31(m,3H),7.06–6.94(m,2H),4.47(s,2H),2.88–2.72(m,1H),2.07(d,J＝12.4Hz,2H),1.93–1.80(m,2H),1.61(dt,J＝13.7,11.0Hz,2H),1.52–1.35(m,2H).MS(ESI)m/z:calculated for C24H22Cl2N4O3[M+H]+:485.11,found:485.11.。
实施例61
(4-(5-(5-氯-1H-吲哚-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)(3-(4-氯苯基)-1,2,4-恶二唑-5-基)甲酮(Ⅱ-16)的合成
原料为实施例47步骤2的2-(5-氯-1H-吲哚-2-基)-5-(哌啶-4-基)-1,3,4-恶二唑、实施例47步骤3的3-(4-氯苯基)-1,2,4-恶二唑-5-羧酸
参照实施例1中步骤3所述的方法,得到白色固体22mg，产率69.84％。1H NMR(400MHz,DMSO-d6)δ12.67–12.34(m,1H),8.14–8.04(m,2H),7.75(d,J＝2.1Hz,1H),7.73–7.66(m,2H),7.49(d,J＝8.7Hz,1H),7.26(dt,J＝8.6,2.3Hz,1H),7.18(dd,J＝2.2,0.9Hz,1H),4.42(d,J＝13.3Hz,1H),4.22(d,J＝13.6Hz,1H),3.56(ddt,J＝14.7,10.7,5.4Hz,2H),2.37–2.16(m,2H),1.98–1.83(m,2H),0.86(t,J＝6.2Hz,1H).MS(ESI)m/z:calculated for C24H18Cl2N6O3[M+H]+:509.08,found:509.53.。
实施例62
中间体2-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)-2-氧代乙酰酰肼的合成
步骤1：2-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)-2-氧代乙酸乙酯的合成
原料为实施例39步骤3的2-(5-氯苯并呋喃-2-基)-5-(哌啶-4-基)-1,3,4-恶二唑、草酰氯单乙酯。参照实施例3步骤2所述方法，得白色固体217mg，产率92.7％。1H NMR(300MHz,Chloroform-d)δ7.70(dd,J＝2.1,0.6Hz,1H),7.57(dt,J＝8.8,0.7Hz,1H),7.47(d,J＝0.9Hz,1H),7.43(dd,J＝8.9,2.1Hz,1H),4.48–4.32(m,3H),3.85(dtd,J＝13.9,4.4,1.3Hz,1H),3.38(dddd,J＝13.8,10.4,5.4,3.6Hz,2H),3.18(ddd,J＝13.9,10.7,3.3Hz,1H),2.36–2.20(m,2H),2.06(ddtd,J＝24.3,14.4,10.5,4.1Hz,2H),1.40(t,J＝7.1Hz,3H).
步骤2：2-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)-2-氧代乙酰酰肼的合成。
参照实施例3步骤1所述方法，于95℃反应。得白色固体154mg，产率72.17％。MS(ESI)m/z:calculated for C17H16ClN5O4[M+H]+:390.09,found:390.21。
实施例63
中间体(1s，3s)-3-(三氟甲氧基)环丁烷-1-甲酸的合成
步骤1：苄基(1s，3s)-3-羟基环丁烷-1-甲酸酯的合成
用10ml无水甲醇将3-氧代环丁烷-1-甲酸苄酯(436mg，2.13mmol)溶解，在-30℃下，逐份添加NaBH4(85mg 2.24mmol)，反应3h。添加NH4Cl饱和溶液，将滤液浓缩除去挥发物。用EA萃取，收集有机相，浓缩得无色透明液体407mg，产率92.33％。1H NMR(300MHz,Chloroform-d)δ7.45–7.33(m,5H),4.27–4.14(m,1H),2.75–2.55(m,3H),2.48–2.32(s,1H),2.26–2.15(m,2H).
步骤2：苄基(1s，3s)-3-(三氟甲氧基)环丁烷-1-甲酸酯的合成
于包裹由锡纸并冷却在-35℃的三颈烧瓶中，依次添加AgOTf(I)(7.40g，28.80mmol)、Selectfluor(5.10g，14.40mmol)、KF(2.23g，38.40mmol)、以及溶于无水乙酸乙酯的苄基(1s，3s)-3-羟基环丁烷-1-甲酸酯(1.98g，9.60mmol)，N2保护下，放置30min。后依次滴加2-氟吡啶(2.54ml，28.80mmol)、TMSCF3(4.26ml，28.80mmol)保证内部温度低于-30℃。随后移入室温隔夜反应。将反应液经硅藻土过滤，EA洗涤，浓缩溶液，残留物经硅胶柱层析(PE/EA＝200/1)洗脱，得无色透明液体1.30g，产率49.38％。1H NMR(300MHz,Chloroform-d)δ7.38(d,J＝14.1Hz,5H),5.17(d,J＝2.1Hz,2H),4.59(t,J＝7.6Hz,1H),2.85–2.48(m,5H).19F NMR(282MHz,Chloroform-d)δ-59.55.MS(ESI)m/z:calculated for C13H13F3O3[M+Na]+:297.24,found:297.11
步骤3:(1s，3s)-3-(三氟甲氧基)环丁烷-1-甲酸的合成
苄基(1s，3s)-3-(三氟甲氧基)环丁烷-1-甲酸酯(1.21g，4.40mmol)用THF溶解，加入NaOH(880mg，22mmol)隔夜反应。将反应也浓缩，残留物用二氯甲烷/1molHCl水溶液萃取，收集有机相，旋干得无色液体793mg，产率97.94％。1H NMR(400MHz,DMSO-d6)δ12.63(s,1H),4.47(p,J＝7.1Hz,1H),3.95(s,2H),3.81-3.70(m,1H),2.75(tdt,J＝9.0,5.7,2.4Hz,2H),2.15-2.03(m,2H).。
实施例64
(4-(5-(5-(氯代呋喃-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)(5-((1s，3s)-3-(三氟甲氧基)环丁 基)-1,3,4恶二唑-1-基)甲酮(Ⅱ-17)的合成
步骤1：(1s，3s)-N'-(2-(4-(5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)-2-氧代乙酰基)-3-(三氟甲氧基)环丁烷-1-甲酰肼的合成
参照实施例1中步骤3所述的方法,得到白色固体157mg，产率73.21％.1H NMR(300MHz,DMSO-d6)δ10.16(s,2H),7.91(d,J＝2.2Hz,1H),7.84(d,J＝8.9Hz,1H),7.78(s,1H),7.54(dd,J＝8.9,2.2Hz,1H),4.81(p,J＝7.6Hz,1H),4.23(d,J＝13.0Hz,2H),4.13–3.95(m,2H),3.06(t,J＝12.0Hz,2H),2.78–2.69(m,1H),2.30(q,J＝9.6Hz,2H),2.15(d,J＝13.1Hz,2H),1.83(dq,J＝25.7,11.7,10.7Hz,3H).
步骤2：(4-(5-(5-(氯代呋喃-2-基)-1,3,4-恶二唑-2-基)哌啶-1-基)(5-((1s，3s)-3-(三氟甲氧基)环丁基)-1,3,4恶二唑-1-基)甲酮的合成
参照实施例39中步骤3的合成得白色固体25mg，产率27.13％。1H NMR(400MHz,Chloroform-d)δ7.70(d,J＝2.1Hz,1H),7.57(d,J＝8.8Hz,1H),7.48(s,1H),7.43(dd,J＝8.8,2.1Hz,1H),5.00–4.91(m,1H),4.76(p,J＝7.6Hz,1H),4.62(dt,J＝13.8,4.4Hz,1H),3.71(ddd,J＝13.9,10.7,3.1Hz,1H),3.44(tdd,J＝10.1,7.5,5.2Hz,2H),3.34(ddd,J＝13.8,10.7,3.2Hz,1H),2.94(dddt,J＝9.9,7.4,5.3,2.6Hz,2H),2.80(tdd,J＝10.3,8.0,2.8Hz,2H),2.34(dt,J＝12.6,3.9Hz,2H),2.19–2.06(m,2H).MS(ESI)m/z:calculated for C23H19ClF3N5O5[M+H]+:528.10,found:528.88.。
实施例65
中间体5-氯-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)苯并呋喃-2-甲酰胺的合成
步骤1：5-氯-N-(哌啶-4-基)苯并呋喃-2-甲酰胺的合成
原料为实施例1步骤2所得的5-氯苯呋喃-2-羧酸、4-氨基哌啶-1-甲酸叔丁酯。参照实施例1步骤3的合成。后经1.5ml三氟乙酸脱Boc，得白色酯状固体2.3g，产率84.31％。1H NMR(300MHz,DMSO-d6)δ8.92(d,J＝7.6Hz,1H),8.68(s,1H),7.89(d,J＝2.2Hz,1H),7.71(d,J＝8.8Hz,1H),7.58(d,J＝1.0Hz,1H),7.49(dd,J＝8.8,2.2Hz,1H),4.15(d,J＝5.2Hz,1H),4.12–4.02(m,1H),3.32(s,1H),3.03(td,J＝12.6,3.1Hz,2H),1.97(dd,J＝13.8,3.9Hz,2H),1.86–1.72(m,2H).
步骤2：2-(4-(5-氯苯呋喃-2-甲酰胺)哌啶-1-基)-2-氧代乙酸乙酯的合成
参照实施例3步骤2所述方法，得白色固体1.74g，产率72.3％。1H NMR(400MHz,Chloroform-d)δ7.65(d,J＝2.0Hz,1H),7.44(d,J＝8.6Hz,2H),7.38(dd,J＝8.8,2.1Hz,1H),6.69(d,J＝8.1Hz,1H),4.54(ddt,J＝13.5,4.5,2.6Hz,1H),4.35(q,J＝7.1Hz,2H),4.28(ddt,J＝11.7,8.2,4.0Hz,1H),3.75(ddt,J＝13.8,5.1,2.8Hz,1H),3.28(ddd,J＝14.1,11.9,2.8Hz,1H),2.93(ddd,J＝13.5,12.0,3.0Hz,1H),2.27–2.09(m,2H),1.71–1.51(m,2H),1.37(t,J＝7.1Hz,3H).
步骤3：5-氯-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)苯并呋喃-2-甲酰胺的合成
参照实施例3步骤1所述方法，于85℃反应。得白色固体543mg，产率60.17％。1H NMR(400MHz,DMSO-d6)δ9.77(s,1H),8.75(d,J＝7.9Hz,1H),7.88(d,J＝2.3Hz,1H),7.70(d,J＝8.8Hz,1H),7.55(s,1H),7.49(dd,J＝9.0,2.3Hz,1H),4.51(d,J＝23.5Hz,1H),4.43(s,1H),4.27(d,J＝13.3Hz,1H),4.17–4.02(m,1H),3.79(d,J＝13.5Hz,1H),3.21–3.13(m,1H),2.90–2.78 (m,1H),1.85(d,J＝11.9Hz,2H),1.63–1.42(m,2H).。
实施例66
5-氯-N-(1-(5-((1s，3s)-3-(三氟甲氧基)环丁基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)苯并呋喃-2-甲酰胺(Ⅰ-29)的合成
步骤1：5-氯-N-(1-(2-氧代-2-(2-((1s，3s)-3-(三氟甲氧基)环丁烷-1-羰基)肼基)乙酰基)哌啶-4-基)苯并呋喃-2-甲酰胺的合成
原料为实施例65步骤3所得的5-氯-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)苯并呋喃-2-甲酰胺、实施例63步骤3所得(1s，3s)-3-(三氟甲氧基)环丁烷-1-甲酸。参照实施例1步骤3的合成，得白色固体203mg，产率54.89％。1H NMR(400MHz,DMSO-d6)δ10.05(s,1H),8.79(t,J＝7.5Hz,2H),7.93–7.89(m,1H),7.70(s,1H),7.55–7.51(m,1H),7.48(d,J＝2.3Hz,1H),4.82(p,J＝7.6Hz,1H),4.31(d,J＝14.7Hz,1H),4.12(dd,J＝7.4,3.8Hz,1H),3.98(d,J＝13.3Hz,1H),2.98–2.81(m,2H),2.72(tt,J＝9.4,7.5Hz,1H),2.60–2.52(m,2H),2.36–2.19(m,2H),1.57(dddt,J＝37.1,19.3,11.4,6.0Hz,4H).
步骤2：5-氯-N-(1-(5-((1s，3s)-3-(三氟甲氧基)环丁基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)苯并呋喃-2-甲酰胺的合成
参照实施例39中步骤3的合成得白色固体26.3mg，产率21.07％。1H NMR(400MHz,Chloroform-d)δ7.66(d,J＝2.0Hz,1H),7.42(s,2H),7.38(dd,J＝9.0,2.1Hz,1H),6.63(d,J＝8.1Hz,1H),4.98(dq,J＝14.1,3.3Hz,1H),4.80–4.68(m,2H),4.35(tdt,J＝11.6,8.2,4.2Hz,1H),3.53–3.36(m,2H),3.16–3.02(m,1H),2.91(dddt,J＝9.9,7.4,5.3,2.7Hz,2H),2.84–2.70(m,2H),2.32–2.17(m,2H),1.79–1.57(m,2H).MS(ESI)m/z:calculated for C22H20ClF3N4O5[M+H]+:513.87,found:513.81.。
实施例67
中间体5-氯-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)-1H-吲哚-2-甲酰胺的合成
步骤1：5-氯-N-(哌啶-4-基)-1H-吲哚-2-甲酰胺的合成
原料为5-氯-1H-吲哚-2-羧酸、4-氨基哌啶-1-甲酸叔丁酯。参照实施例65步骤1的合成，得白色固体1.56g。产率72.14％。1H NMR(400MHz,DMSO-d6)δ11.82(d,J＝2.2Hz,1H),8.77(s,1H),8.59(d,J＝7.6Hz,1H),7.71(d,J＝2.0Hz,1H),7.44(d,J＝8.7Hz,1H),7.23–7.16(m,2H),4.09(dtt,J＝10.3,7.0,3.4Hz,1H),3.38–3.31(m,2H),3.12–2.97(m,2H),2.06–1.95(m,2H),1.82–1.66(m,2H).
步骤2：2-(4-(5-氯-1H-吲哚-2-甲酰胺)哌啶-1-基)-2-氧代乙酸乙酯的合成
参照实施例3步骤2所述方法，得白色固体1.23g，产率43.21％。1H NMR(400MHz,DMSO-d6)δ11.81(d,J＝2.2Hz,1H),8.45(d,J＝8.0Hz,1H),7.71(d,J＝2.0Hz,1H),7.43(d,J＝8.7Hz,1H),7.18(dd,J＝8.7,2.1Hz,1H),7.14(dd,J＝2.1,0.9Hz,1H),4.31(qd,J＝7.1,1.4Hz,2H),4.27–4.20(m,1H),4.15(ddq,J＝11.5,8.0,4.1,3.7Hz,1H),3.62–3.51(m,1H),3.33–3.23(m,1H),2.94(td,J＝12.8,2.9Hz,1H),1.92(dd,J＝13.2,3.9Hz,2H),1.60–1.38(m,2H),1.28(t,J＝7.1Hz,3H).
步骤3：5-氯-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)-1H-吲哚-2-甲酰胺的合成
参照实施例65步骤3的合成，得白色固体431mg，产率46.98％。1H NMR(400MHz, DMSO-d6)δ11.85–11.75(m,1H),9.79(s,1H),8.46(d,J＝7.6Hz,1H),7.70(d,J＝2.1Hz,1H),7.43(d,J＝8.7Hz,1H),7.18(dd,J＝8.8,2.1Hz,1H),7.16(d,J＝1.6Hz,1H),4.50(d,J＝47.4Hz,2H),4.27(d,J＝13.2Hz,1H),4.18–4.03(m,1H),3.78(d,J＝13.9Hz,1H),3.19(ddd,J＝14.1,11.9,2.7Hz,1H),2.87(td,J＝12.7,2.9Hz,1H),1.97–1.81(m,2H),1.50(dqd,J＝39.8,12.4,4.5Hz,2H).。
实施例68
5-氯-N-(1-(5-((1s，3s)-3-(三氟甲氧基)环丁基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)-1H-吲哚-2-甲酰胺(Ⅰ-30)的合成
步骤1：5-氯-N-(1-(2-氧代-2-(2-((1s，3s)-3-(三氟甲氧基)环丁烷-1-羰基)肼基)乙酰基)哌啶-4-基)-1H-吲哚-2-甲酰胺的合成
原料为实施例67步骤3所得的5-氯-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)-1H-吲哚-2-甲酰胺、实施例63步骤3所得(1s，3s)-3-(三氟甲氧基)环丁烷-1-甲酸。参照实施例1步骤3的合成，得白色固体925mg，产率41.11％。1H NMR(400MHz,DMSO-d6)δ11.80(d,J＝2.1Hz,1H),10.58(s,1H),10.04(s,1H),8.45(d,J＝7.8Hz,1H),7.71(d,J＝2.0Hz,1H),7.43(d,J＝8.7Hz,1H),7.18(dd,J＝8.9,2.0Hz,2H),4.82(p,J＝7.6Hz,1H),4.27(d,J＝13.2Hz,1H),4.14(pt,J＝7.2,3.6Hz,1H),3.98(d,J＝13.4Hz,1H),3.31–3.20(m,1H),2.91(td,J＝12.9,3.0Hz,1H),2.72(tt,J＝9.4,7.5Hz,1H),2.60–2.51(m,2H),2.36–2.21(m,2H),1.91(dt,J＝13.2,7.3Hz,2H),1.58(qd,J＝12.1,4.2Hz,1H),1.46(qd,J＝12.3,4.3Hz,1H).
步骤2：5-氯-N-(1-(5-((1s，3s)-3-(三氟甲氧基)环丁基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)-1H-吲哚-2-甲酰胺的合成(Ⅰ-30)
参照实施例66步骤2的合成，得白色固体57mg，产率58.67％。1H NMR(400MHz,DMSO-d6)δ8.45(d,J＝7.8Hz,1H),7.71(d,J＝2.0Hz,1H),7.43(d,J＝8.7Hz,1H),7.18(dd,J＝8.9,2.0Hz,2H),4.82(p,J＝7.6Hz,1H),4.27(d,J＝13.2Hz,1H),4.14(pt,J＝7.2,3.6Hz,1H),3.98(d,J＝13.4Hz,1H),3.31–3.20(m,1H),2.91(td,J＝12.9,3.0Hz,1H),2.72(tt,J＝9.4,7.5Hz,1H),2.60–2.51(m,2H),2.36–2.21(m,2H),1.91(dt,J＝13.2,7.3Hz,2H),1.58(qd,J＝12.1,4.2Hz,1H),1.46(qd,J＝12.3,4.3Hz,1H).MS(ESI)m/z:calculated for C22H21ClF3N5O4[M+H]+:511.89,found:511.74.。
实施例69
中间体2-((1s，3s)-3-(三氟甲氧基)环丁氧基)乙酸的合成
步骤1：(1s，3s)-3-(苄氧基)环丁烷-1-醇的合成
参照实施例63步骤1的合成，得无色液体5.40g，产率91.37％。1H NMR(500MHz,Chloroform-d)δ7.42–7.34(m,2H),7.32–7.22(m,3H),4.60(t,J＝1.0Hz,2H),3.99(d,J＝4.9Hz,1H),3.88(s,1H),3.66(d,J＝4.9Hz,1H),1.98–1.85(m,3H).
步骤2：2-(3-对-(苄氧基)环丁氧基)乙酸叔丁酯的合成
原料为实施例63步骤1所得3-顺-(苄氧基)环丁醇，2-氯乙酸叔丁酯。将3-顺-(苄氧基)环 丁醇(1.30g，7.29mmol)、CsCO3(2.81g,14.59mmol)溶于DMF(10ml),10min后加入2-氯乙酸叔丁酯(1.10g,7.29mmol),隔夜反应。用乙酸乙酯萃取，无水硫酸钠干燥，过滤，滤液经减压浓缩，残留物通过硅胶柱层析纯化得到无色液体2.10g，产率98.47％。1H NMR(400MHz,Chloroform-d)δ7.38-7.27(m,5H),4.42(s,2H),3.89(s,2H),3.75-3.62(m,2H),2.65(m,1H),2.09-1.99(s,4H),1.47(s,9H).
步骤3：2-(3-顺-羟基环丁氧基)乙酸叔丁酯的合成
N2保护,Pd/C(230mg,10％Pd/C)加入到2-(3-对-(苄氧基)环丁氧基)乙酸叔丁酯(2g,684mmol)的MeOH(25ml)。H2下，于50℃下搅拌5h。将反应物经硅藻土过滤，滤液旋干得淡黄色液体1.27g，产率91.80％。1H NMR(400MHz,Chloroform-d)δ3.95-3.89(m,1H),3.89(s,2H),3.71-1.62(m,1H),2.77-2.67(m,2H),2.00-1.91(m,2H),1.80(s,1H),1.47(s,9H).
步骤4：2-(3-氯-(三氟甲氧基)环丁氧基)乙酸叔丁酯的合成
参照实施例63步骤2的合成，得无色液体1.01g，产率41.22％。1H NMR(300MHz,Chloroform-d)δ4.36-4.26(m,1H),3.89(s,2H),3.82-3.73(m,1H),2.86-2.70(m,2H),2.37-2.28(m,2H),1.47(s,9H).
步骤5：2-((1s，3s)-3-(三氟甲氧基)环丁氧基)乙酸的合成
将2-(3-氯-(三氟甲氧基)环丁氧基)乙酸叔丁酯(1.01g)用DCM(8ml)溶解，加入TFA 2ml，隔夜反应。将反应液浓缩，无需纯化直接用于下一步。得淡黄色液体800mg，产率100％。1NMR(400MHz,Chloroform-d):δ4.26-4.19(m,1H),4.00(s,2H),3.73-3.70(m,1H),2.77-2.74(m,2H),2.27-2.24(m,2H).。
实施例70
N-((1r，4R)-4-(5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)环己基)-2-(((1s，3S)-3-(三氟甲氧基)环丁氧基)乙酰胺(Ⅱ-18)的合成
原料为实施例40步骤2所得的(1r，4r)-4-(5-(5-氯苯并呋喃-2-基)-1,3,4-恶二唑-2-基)环己烷-1-胺、实施例69步骤5所得的2-((1s，3s)-3-(三氟甲氧基)环丁氧基)乙酸。参照实施例1步骤3的合成，得白色固体34mg，产率41.22％。1H NMR(300MHz,DMSO-d6)δ(ppm):8.07(d,J＝8.2Hz,1H),7.90(s,1H),7.84(d,J＝8.9Hz,1H),7.77(d,J＝6.9Hz,1H),7.54(d,J＝9.0Hz,1H),4.49(s,2H),4.26-4.19(m,1H),3.72(d,J＝9.2Hz,1H),3.72-3.70(m,1H),3.11-2.93(m,3H),2.19(d,J＝13.5Hz,2H),2.27-2.24(m,2H),1.91(d,J＝12.3Hz,2H),1.67(d,J＝12.4Hz,2H),1.47(q,J＝12.6Hz,2H)；MS(ESI)m/z:calculated for C23H23ClF3N3O5[M+H]+:514.90,found:514.56.。
实施例71
中间体6-溴-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)喹啉-3-甲酰胺的合成
步骤1：6-溴-N-(哌啶-4-基)喹啉-3-甲酰胺的合成
参照实施例65步骤1的合成，得黄白色固体1.02g。产率74.15％。1H NMR(300MHz,Chloroform-d)δ8.34(d,J＝8.5Hz,1H),8.23(d,J＝8.5Hz,1H),8.16(d,J＝8.5Hz,1H),8.06(d,J＝2.2Hz,1H),8.02(d,J＝9.0Hz,1H),7.85(dd,J＝9.0,2.2Hz,1H),4.16(dddd,J＝15.1,10.9,8.4,4.1Hz,1H),3.23(dt,J＝12.7,3.7Hz,2H),2.85(td,J＝11.9,2.6Hz,2H),2.13(dd,J＝13.1,3.7Hz,2H),1.75–1.54(m,2H).
步骤2：2-(4-(6-溴喹啉-3-甲酰胺)哌啶-1-基)-2-氧代乙酸乙酯的合成
参照实施例65步骤2的合成，得白色固体1.3g，产率87.88％。1H NMR(300MHz,Chloroform-d)δ8.33(d,J＝8.5Hz,1H),8.29–8.23(m,1H),8.19(d,J＝8.4Hz,1H),8.07(d,J＝2.1Hz,1H),8.00(d,J＝9.0Hz,1H),7.86(dd,J＝9.0,2.2Hz,1H),4.56(dd,J＝11.6,6.9Hz,1H),4.43–4.25(m,3H),3.86–3.70(m,1H),3.32(ddd,J＝14.2,11.8,2.8Hz,1H),3.00(ddd,J＝13.6,11.8,3.0Hz,1H),2.30–2.13(m,2H),1.71(pd,J＝11.9,4.3Hz,2H),1.40(t,J＝7.1Hz,3H).
步骤3：6-溴-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)喹啉-3-甲酰胺的合成
参照实施例65步骤3的合成，得白色固体534mg，产率78.06％。1H NMR(400MHz,DMSO-d6)δ9.43(d,J＝274.5Hz,1H),8.86(d,J＝8.4Hz,1H),8.54(d,J＝8.6Hz,1H),8.41(d,J＝2.2Hz,1H),8.19(d,J＝8.5Hz,1H),8.09(d,J＝9.0Hz,1H),7.99(dd,J＝9.0,2.2Hz,1H),4.62–4.38(m,2H),4.30(d,J＝13.2Hz,1H),4.15(dddd,J＝15.8,10.5,8.0,4.7Hz,1H),3.82(d,J＝13.6Hz,1H),3.27–3.13(m,1H),2.86(td,J＝13.0,3.1Hz,1H),1.90(ddd,J＝12.3,8.1,3.9Hz,2H),1.65(dqd,J＝28.6,12.3,4.2Hz,2H).。
实施例72
6-溴-N-(1-(5-((1s，3s)-3-(三氟甲氧基)环丁基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)喹啉-3-甲酰胺(Ⅰ-31)的合成
步骤1：6-溴-N-(1-(2-氧代-2-(2-((1s，3s)-3-(三氟甲氧基)环丁烷-1-羰基)肼基)乙酰基)哌啶-4-基)喹啉-3-甲酰胺的合成
参照实施例66步骤1的合成，得白色固体347mg，产率53.25％。1H NMR(400MHz,DMSO-d6)δ10.42(d,J＝105.6Hz,1H),10.05(s,1H),8.85(d,J＝8.3Hz,1H),8.55(d,J＝8.5Hz,1H),8.41(d,J＝2.2Hz,1H),8.19(d,J＝8.5Hz,1H),8.09(d,J＝9.0Hz,1H),7.99(dd,J＝9.0,2.3Hz,1H),4.82(p,J＝7.6Hz,1H),4.30(d,J＝13.2Hz,1H),4.25–4.09(m,1H),4.02(t,J＝10.6Hz,1H),3.27(t,J＝12.5Hz,1H),2.90(t,J＝12.3Hz,1H),2.72(tt,J＝9.2,7.4Hz,1H),2.60–2.51(m,2H),2.38–2.19(m,2H),1.92(d,J＝7.4Hz,2H),1.68(dqd,J＝41.3,12.2,4.2Hz,2H).
步骤2：6-溴-N-(1-(5-((1s，3s)-3-(三氟甲氧基)环丁基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)喹啉-3-甲酰胺的合成
参照实施例66步骤2的合成，得白色固体40mg，产率22.51％。1H NMR(400MHz,Chloroform-d)δ8.32(d,J＝8.4Hz,1H),8.23(d,J＝8.6Hz,1H),8.20(d,J＝8.2Hz,1H),8.05(s,1H),7.97(d,J＝8.9Hz,1H),7.84(d,J＝9.1Hz,1H),5.08–4.90(m,1H),4.74(p,J＝7.0,6.1Hz,2H),4.37(ddp,J＝11.8,8.5,4.2Hz,1H),3.63–3.35(m,2H),3.25–3.05(m,1H),3.02–2.86(m,2H),2.78(q,J＝10.2Hz,2H),2.26(dt,J＝12.9,4.0Hz,2H),1.77(dqd,J＝24.6,12.0,4.2Hz,2H).MS(ESI)m/z:calculated for C23H21BrF3N5O4[M+H]+:569.35,found:569.21.。
实施例73
中间体2-(三氟甲氧基)乙烷-1-醇的合成
步骤1：(2-(三氟甲氧基)乙氧基)甲基)苯的合成
原料为2-苄氧基乙醇。参照实施例63步骤2的合成得无色液体1.37g，产率45.76％。1H NMR(300MHz,Chloroform-d)δ7.31(s,5H),4.60(s,2H),4.16-4.10(m,2H),3.74-3.67(m,2H).
步骤2：2-(三氟甲氧基)乙烷-1-醇的合成
参照实施例69步骤3的无色液体782mg，产率：96.63％。1H NMR(300MHz,Chloroform-d)δ4.16-4.10(m,2H),3.74(t,J＝4.5Hz,2H).。
实施例74
6-溴-N-(1-(5-(2-(三氟甲氧基)乙氧基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)喹啉-3-甲酰胺(Ⅰ-32)的合成
步骤1：2-(三氟甲氧基)乙基-2-(2-(4-(6-溴喹啉-3-甲酰胺)哌啶-1-基)-2-氧代乙酰基)肼-1-羧酸酯得合成
在0℃下，将吡啶(134mg,1.69mmol)投入2-(三氟甲氧基)乙烷-1-醇(146mg,1.13mmol)和三光气(67mg，225.48μmol)的THF(3ml)中，搅拌60min后投入6-溴-N-(1-(2-肼基-2-氧代乙酰基)哌啶-4-基)喹啉-3-甲酰胺(327mg，567.39μmol)，室温隔夜反应。将反应液用EA/H2O萃取，收集EA相。EA相用Na2SO4干燥，抽滤，滤液浓缩，残留物通过硅胶柱层析纯化得白色固体134mg，产率46.53％。1H NMR(400MHz,DMSO-d6)δ10.42(d,J＝105.6Hz,1H),10.05(s,1H),8.85(d,J＝8.3Hz,1H),8.55(d,J＝8.5Hz,1H),8.41(d,J＝2.2Hz,1H),8.19(d,J＝8.5Hz,1H),8.09(d,J＝9.0Hz,1H),7.99(dd,J＝9.0,2.3Hz,1H),4.30(d,J＝13.2Hz,1H),4.25-4.09(m,3H),4.02(t,J＝10.6Hz,1H),4.00-3.96(m,2H),3.27(t,J＝12.5Hz,1H),2.90(t,J＝12.3Hz,1H),1.92(d,J＝7.4Hz,2H),1.68(dqd,J＝41.3,12.2,4.2Hz,2H).
步骤2：6-溴-N-(1-(5-(2-(三氟甲氧基)乙氧基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)喹啉-3-甲酰胺的合成
参照实施例66步骤2的合成，得白色固体27mg，产率32.11％。1H NMR(300MHz,Chloroform-d)δ8.33(d,J＝8.5Hz,1H),8.29–8.23(m,1H),8.19(d,J＝8.4Hz,1H),8.07(d,J＝2.1Hz,1H),8.00(d,J＝9.0Hz,1H),7.86(dd,J＝9.0,2.2Hz,1H),4.40(s,2H),4.56(dd,J＝11.6,6.9Hz,1H),4.43–4.25(m,3H),3.98(d,J＝12.3Hz,1H),3.86–3.70(m,1H),3.82(d,J＝12.5Hz,1H),3.32(ddd,J＝14.2,11.8,2.8Hz,1H),3.00(ddd,J＝13.6,11.8,3.0Hz,1H),2.30–2.13(m,2H),1.71(pd,J＝11.9,4.3Hz,2H),1.40(t,J＝7.1Hz,3H).MS(ESI)m/z:calculated for C21H19BrF3N5O5[M+H]+:558.31,found:558.75.。
实施例75
5-氯-N-(1-(5-(2-(三氟甲氧基)乙氧基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)苯并呋喃-2-甲酰胺(Ⅰ-33)的合成
步骤1：2-(三氟甲氧基)乙基-2-(2-(4-(5-氯苯并呋喃-2-甲酰胺)哌啶-1-基)-2-氧代乙酰基)肼-1-羧酸盐的合成
参照实施例74步骤1的合成，得白色固体178mg，产率：50.83％。1H NMR(300MHz,DMSO-d6)δ11.27(s,1H),9.49(s,1H),8.92(d,J＝7.6Hz,1H),7.89(d,J＝2.2Hz,1H),7.71(d,J＝8.8Hz,1H),7.58(d,J＝1.0Hz,1H),7.49(dd,J＝8.8,2.2Hz,1H),4.23(s,2H),4.15(d,J＝5.2Hz,1H),4.12–4.02(m,3H),3.32(s,1H),3.03(td,J＝12.6,3.1Hz,2H),1.97(dd,J＝13.8,3.9Hz,2H),1.86–1.72(m,2H).
步骤2：5-氯-N-(1-(5-(2-(三氟甲氧基)乙氧基)-1,3,4-恶二唑-2-羰基)哌啶-4-基)苯并呋喃-2-甲酰胺的合成
参照实施例66步骤2的合成，得白色固体19mg，产率22.36％。1H NMR(400MHz,Chloroform-d)δ7.66(d,J＝2.0Hz,1H),7.42(s,2H),7.38(dd,J＝9.0,2.1Hz,1H),6.63(d,J＝8.1Hz,1H),4.98(dq,J＝14.1,3.3Hz,1H),4.80–4.68(m,1H),4.40-4.35(m,3H),4.02–3.81(m,2H),3.53–3.36(m,2H),2.32–2.17(m,2H),1.79–1.57(m,2H).MS(ESI)m/z:calculated for C20H18ClF3N4O6[M+H]+:503.83,found:503.09.。
实施例76
5-氯-N-(1-(3-(4-氯苯基)-1,2,4-恶二唑-5-羰基)哌啶-4-基)-1H-吲哚-2-甲酰胺(Ⅰ-34)的合成
原料为实施例11步骤3所得的3-(4-氯苯基)-1,2,4-恶二唑-5-羧酸、实施例67步骤1所得的5-氯-N-(哌啶-4-基)-1H-吲哚-2-甲酰胺。参照实施例66步骤1的合成，得白色固体47mg，产率54.71％。1H NMR(400MHz,DMSO-d6)δ11.79(d,J＝2.1Hz,1H),8.46(d,J＝7.8Hz,1H),8.19–7.98(m,2H),7.70(d,J＝2.1Hz,2H),7.68(d,J＝2.0Hz,1H),7.43(dt,J＝8.8,0.7Hz,1H),7.22–7.06(m,2H),5.75(s,1H),4.46(d,J＝13.3Hz,1H),4.27–4.16(m,1H),3.43(td,J＝13.9,12.9,2.8Hz,1H),3.26–3.06(m,2H),2.04(s,1H),1.92(d,J＝12.7Hz,1H),1.63(tdd,J＝23.9,12.1,4.3Hz,2H).MS(ESI)m/z:calculated for C23H19Cl2N5O3[M+H]+:485.34,found:485.08.。
试验例1
ATF4抑制率和IC50的测定
将处于对数生长期的HEK-293T细胞接种于12孔板，贴壁培养12h，加入待测化合物(500nM)作用1h后，再给予毒胡萝卜素(Tg，100nM)，共同孵育3h。弃去原培养基，用PBS缓冲溶液洗涤3次，于冰上加入细胞裂解液，收取蛋白样品，用于SDS-PAGE电泳。后转移到NC膜。室温封闭1h后，ATF4一抗溶液(1:1000)4℃孵育过夜。TBST溶液洗涤3次，HRP标记的二抗溶液室温孵育1h。TBST溶液洗涤3遍后，化学发光器曝光。条带灰度值用Image J软件进行分析。
表1化合物对HEK-293T细胞ATF4抑制率和部分IC50值

由表可以看出本发明中的化合物对于HEK-293T细胞ATF4表达抑制明显。相对于先导化合物ISRIB，化合物Ⅰ-13、Ⅰ-14、Ⅰ-18、Ⅰ-19、Ⅱ-2和Ⅱ-4抑制率活性略优或大致相当。相对于先导化合物ISRIB，化合物Ⅰ-14、Ⅰ-18和Ⅱ-2的ATF4IC50具有较好的数值，显示很好的活性。说明化合物具有激活eIF2B，逆转ISR引起的神经损伤，治疗神经退行性疾病的能力。
试验例2
SDS-PAGE法测蛋白翻译率
将处于对数生长期的CHO细胞接种于6孔板，贴壁培养12h，弃去原培养基，PBS溶液洗涤细胞一次，加入含待测化合物(500nM)的无血清培养基作用1h后，再给予毒胡萝卜素(Tg，100nM)共同孵育24h。在处理24h后，收集细胞上清，并加入PMSF，在2000×g，4℃的条件下离心10min去除细胞碎片；并加入400μL的甲醇，剧烈混匀，并加入200μL的氯仿，剧烈混匀并涡旋，在4℃14000×g离心10min，丢弃上清液；沉淀加入400μL的甲醇混匀，4℃17000×g 10min，弃去上清液，沉淀晾干，加入上样缓冲液，即为上清中蛋白质样品，进行SDS-PAGE电泳，考马斯亮蓝染色，再用脱色液洗至背景干净，用化学发光器曝光。
表2化合物对CHO细胞蛋白质翻译影响测定

由表可以看出本发明中的化合物对于CHO细胞蛋白质翻译具有较好的促进作用。相对于先导化合物ISRIB，化合物Ⅰ-2、Ⅰ-9、Ⅰ-10、Ⅰ-11、Ⅰ-15、Ⅰ-16、Ⅰ-17、Ⅰ-25、Ⅰ-26、Ⅰ-28和Ⅱ-5能更好的蛋白质翻译恢复率，并且能接近或超过空白组的数值，说明化合物能通过激活eIF2B明显促进蛋白质合成的恢复，可以逆转ISR引起的神经损伤，治疗神经退行性疾病。

Claims (8)

  1. 具有式(I)或式(II)结构的化合物，其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，
    其中：
    A为卤素取代的双芳香杂环；
    D选自
    L为
    W为五元杂环；
    n为0、1或2；
    m为0或1；
    B为卤素取代的苯环、
    K选自
    G选自
  2. 根据权利要求1所述的具有式(I)或式(II)结构的化合物，其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，其特征在于：
    A选自
  3. 根据权利要求1所述的具有式(I)或式(II)结构的化合物，其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，其特征在于：
    W选自
  4. 根据权利要求1所述的具有式(I)或式(II)结构的化合物，其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，其特征在于：
    B选自
  5. 选自以下的化合物，其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物，


  6. 一种药物组合物，以权利要求1-5任一项所述化合物、其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物为活性成分或者主要活性成分，辅以药学上可接受的载体组成。
  7. 权利要求1-5任一项所述化合物、其立体异构体、其几何异构体、其互变异构体、其药学上可接受的盐、其前药、其水合物或其溶剂化物在制备治疗和/或预防神经退行性疾病的 药物中的应用。
  8. 根据权利要求7所述的应用，其特征在于：所述神经退行性疾病为阿尔茨海默病、帕金森病、亨廷顿病、肌萎缩性侧索硬化、衰老、孤独症谱系障碍、白质消融性疾病、痴呆或额颞叶型痴呆。
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