WO2023160475A1 - 咪唑并哒嗪类衍生物、其制备方法、药物组合物和用途 - Google Patents

咪唑并哒嗪类衍生物、其制备方法、药物组合物和用途 Download PDF

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WO2023160475A1
WO2023160475A1 PCT/CN2023/076805 CN2023076805W WO2023160475A1 WO 2023160475 A1 WO2023160475 A1 WO 2023160475A1 CN 2023076805 W CN2023076805 W CN 2023076805W WO 2023160475 A1 WO2023160475 A1 WO 2023160475A1
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pain
ethyl
methoxy
bromo
pyridine
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PCT/CN2023/076805
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French (fr)
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王非
陈南阳
孙勇
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上海赛默罗生物科技有限公司
上海赛默罗德生物科技有限公司
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Publication of WO2023160475A1 publication Critical patent/WO2023160475A1/zh

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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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/5025—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with heterocyclic ring systems
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • 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
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • 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
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • 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
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • 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
    • A61P25/06—Antimigraine agents
    • 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
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • the application relates to imidazopyridazine derivatives with regulating function on GABAA receptors, their preparation method, pharmaceutical composition and their application as medicine.
  • GABA ⁇ -Aminobutyric acid
  • GABA A R GABA A receptor
  • GABA B receptor GABA B receptor
  • the GABA A receptor subunits in mammals have been found to include ⁇ 1-6, ⁇ 1-4, ⁇ 1-3, ⁇ , ⁇ , ⁇ and ⁇ 1-2 subunits, among which ⁇ subunit, ⁇ subunit and ⁇ subunit
  • the base pair is essential to form a complete functional GABA A receptor, and the ⁇ subunit is crucial for the binding of benzodiazepines to the GABA A receptor.
  • Drugs that bind at the allosteric binding site can be positive allosteric modulators that increase receptor activity (or positive allosteric modulators), negative allosteric modulators that decrease receptor activity (or inverse allosteric modulators) ) or a neutral allosteric modulator that does not alter receptor activity (this refers to a compound that binds to an allosteric binding site but does not modulate receptor activity).
  • GABA A receptors containing ⁇ 2 or ⁇ 3 subunits may be involved in certain pain states and that positive allosteric modulators of these receptors may be potent Analgesics (Mirza, NR and Munro, G., Drug News and Perspectives, 2010, 23(6), 351-360).
  • the mainstream view is that the regulatory activity of GABA A receptors containing ⁇ 1 subunits is the main source of side effects (such as sedation, addiction, drowsiness, forgetfulness) of current GABA A modulators (such as benzodiazepines) ( Nature Reviews: Drug Discovery by Uwe Rudolph and Frederic Knoflach, 2011, 10(9), 685-697). Finding new compounds that interact with GABA A receptors and have fewer ⁇ 1-GABA A receptor-related side effects would have enormous therapeutic potential.
  • R 1 is a fused group formed by a substituted or unsubstituted heterocyclic ring and a heterocyclic ring;
  • R 2 is selected from H, halogen, OH, C1-C6 alkoxy or CN;
  • R 3 is selected from H, substituted or unsubstituted linear or branched C1-C6 alkyl or substituted or unsubstituted substituted C3-C6 cycloalkyl.
  • substituted means that the designated group or moiety may have 1, 2, 3, 4, 5 or 6 substituents.
  • substituents are independently selected and need not be the same.
  • substituents When indicating the number of substituents, the term "one or more" refers to one substitution up to the highest possible number of substitutions, ie replacement of one hydrogen up to all hydrogens replaced by substituents. Unless otherwise specified, 1, 2, 3, 4 or 5 substituents are preferred.
  • substituted means that one or more hydrogens on the group are selected from C1-C4 alkyl, C1-C4 alkoxy, C1- C4 halogen substituted alkyl or halogen substituted.
  • halogen refers to fluorine, chlorine, bromine and iodine, preferably fluorine.
  • the compounds of the present application may contain asymmetric centers or chiral centers and thus exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the application, including but not limited to, diastereomers, enantiomers and hindered isomers, and mixtures thereof, such as racemic mixtures, will form part of this application.
  • this application refers to all geometric and positional isomers.
  • the compounds of the present application may exist in different tautomeric forms, and all such forms are included within the scope of the present application. All stereoisomers of the compounds of the application are intended to be included in admixture or in pure or substantially pure form. Resolution may be by physical methods such as fractional crystallization, separation or crystallization of diastereomeric derivatives, or separation by chiral column chromatography.
  • prodrug is a functional derivative of a compound of formula (1) which is readily converted into a compound of formula (1) in vivo. Suitable such derivatives can be selected and prepared by conventional techniques well known to those skilled in the art, see for example Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.
  • pharmaceutically acceptable salt refers to a pharmaceutically acceptable organic or inorganic salt of the compound of the present application.
  • Exemplary salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, acid sulfate, isonicotinate, lactate , salicylate, acid citrate, succinate, maleate, fumarate, gluconate, formate, mesylate and pamolate.
  • “Pharmaceutically acceptable salt” may refer to the inclusion of another molecule such as maleate or other counterion. The counterion stabilizes the charge in the parent compound.
  • a "pharmaceutically acceptable salt” can have more than one charged atom, and multiple charged atoms can have multiple counterions.
  • the desired “pharmaceutically acceptable salt” can be prepared by a suitable method, for example, treating the free base with the following inorganic acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid; or The following organic acids: acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, salicylic acid, pyranosidic acids such as glucuronic acid or galacturonic acid, alpha-hydroxy Acids such as citric acid or tartaric acid, amino acids such as glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, sulfonic acids such as methanesulfonic acid or p-toluenesulfonic acid.
  • inorganic acids hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid
  • organic acids acetic acid, maleic acid, succinic acid, mandelic acid,
  • the desired "pharmaceutically acceptable salt” can be prepared by suitable methods, for example, by treating the free acid with an inorganic or organic base as follows: amine, alkali metal hydroxide or alkaline earth metal hydrogen oxides, etc.
  • suitable salts include, but are not limited to, organic salts derived from amino acids, salts of primary, secondary and tertiary amines, and salts of cyclic amines such as piperidine, morpholine and piperazine, and salts derived from sodium, calcium, potassium , magnesium, manganese, iron, copper, zinc, aluminum and lithium derived inorganic salts.
  • the compounds of the present application can exist in a continuum of solid states from fully amorphous to fully crystalline.
  • amorphous refers to a state in which a material lacks long-range order at the molecular level and, depending on temperature, can exhibit the physical properties of a solid or a liquid. Typically, such materials do not give significant X-ray diffraction patterns and, while exhibiting solid properties, are more formally described as liquids.
  • a change from solid properties to liquid properties occurs, characterized by a change of state, usually second order ("glass transition").
  • crystal refers to a solid phase in which the material has a regular and ordered internal structure at the molecular level and gives a distinctive X-ray diffraction pattern with defined peaks. Such materials will also exhibit the properties of liquids when heated sufficiently, but the change from solid to liquid is characterized by a phase transition, usually first order ("melting point").
  • polymorph refers to different solid crystalline phases produced by certain compounds of the present application due to the existence of two or more different molecular arrangements in the solid state. Certain compounds of the present application may exist in more than one crystalline form, and this application is intended to include each crystalline form and mixtures thereof.
  • solvate refers to a combination or complex of one or more solvent molecules with a compound of the present application.
  • solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethylsulfoxide, ethyl acetate, acetic acid, and ethanolamine.
  • the compounds of the present application can exist in unsolvated form, and can also exist in solvated form with pharmaceutically acceptable solvents such as water, ethanol, etc., so the present application will include solvated and unsolvated forms.
  • the compounds of the present application may contain unnatural proportions of atomic isotopes at one or more of the atoms that make up the compound, the term "isotopologue" having the same atomic number but different atomic mass or mass number than that which predominates in nature The atomic mass or mass number present.
  • compounds can be labeled with radioactive isotopes, such as deuterium (2H), tritium (3H), iodine-125 (125I), or C-14 (14C). All changes in isotopic composition of the compounds of the present application, whether radioactive or not, are included within the scope of the present application.
  • Isotopic variants may confer certain therapeutic advantages, such as deuterium enrichment to increase in vivo half-life or reduce dosage requirements, or may provide standard compounds that can be used for characterization of biological samples. can be prepared without undue experimentation by conventional techniques well known to those skilled in the art, or by methods analogous to those described in the Schemes and Examples herein, using appropriate isotopically enriched reagents and/or intermediates Isotopically enriched compounds within formula (1).
  • the R 2 is selected from H, halogen, OH, C1-C6 alkoxy or CN; preferably, the R 2 is preferably H or halogen, more preferably H or F.
  • the R 3 is selected from H, substituted or unsubstituted linear or branched C1-C6 alkyl or substituted or unsubstituted C3-C6 cycloalkyl.
  • the C1-C6 alkyl group can be a straight chain or a branched chain.
  • Exemplary C1-C6 alkyl groups include methyl, ethyl, n-propyl (1-propyl), isopropyl Base (2-propyl, 1-methylethyl), n-butyl (1-butyl), sec-butyl (2-butyl, 1-methylpropyl), isobutyl (2-methyl Propyl) or Fe/F-butyl (1,1-dimethylethyl).
  • One or more hydrogen atoms on the above-mentioned alkyl groups may be substituted by methyl, ethyl, n-propyl, methoxy, ethoxy, propoxy, fluoromethyl, fluoroethyl, fluorine, chlorine, bromine, etc. base replaced.
  • cycloalkyl refers to a monovalent saturated cyclic hydrocarbon group.
  • exemplary C3-C5 cycloalkyl groups include cyclopropyl, cyclobutyl or cyclopentyl.
  • exemplary C3-C6 cycloalkyl groups include 1-methylcyclopropyl, 2-methylcyclopropyl, 1-methylcyclobutyl, 2-methylcyclobutyl, 3-methyl Cyclobutyl, 1-methylcyclopentyl, 2-methylcyclopentyl or 3-methylcyclopentyl.
  • One or more hydrogen atoms on the cycloalkyl group can be replaced by methyl, ethyl, n-propyl, methoxy, ethoxy, propoxy, fluoromethyl, fluoroethyl, fluorine, chlorine, bromine, etc. Substituents are substituted.
  • alkoxy refers to an alkyloxy group
  • exemplary C1-C6 alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy , sec-butoxy, etc.
  • One or more hydrogen atoms on the above-mentioned alkoxy groups can be replaced by methyl, ethyl, n-propyl, methoxy, ethoxy, propoxy, fluoromethyl, fluoroethyl, fluorine, chlorine, bromine, etc. Substituents are substituted.
  • R is selected from linear or branched C1-C6 alkyl, C1-C6 alkoxy, more preferably C2-C4 alkyl, more preferably ethyl or isopropyl base.
  • the R 1 is a heterocycle and a heterocycle fused group, and the two heterocycles independently refer to a saturated or partially unsaturated monocyclic or polycyclic group with heteroatoms.
  • the heteroatom is N, O or S.
  • the two heterocycles are independently 3-7 membered saturated or partially unsaturated monocyclic or polycyclic groups containing 1-3 ring heteroatoms selected from N, O or S.
  • S and O can be present as -SO or SO2 .
  • the two heterocycles are independently 5-7 membered saturated or unsaturated monocyclic groups containing 1, 2 or 3 ring heteroatoms selected from N, O or S.
  • the two heterocycles are independently 5-6 membered saturated or unsaturated monocyclic groups containing 1, 2 or 3 ring heteroatoms selected from N or O.
  • R has a structure shown in formula (2), (3), (4), (5), (6) or (7):
  • the R is selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxyl, C1-C6 alkylthio or C1-C6 alkylsulfonyl; above-mentioned groups (including related Alkyl, cycloalkyl, alkoxy, alkylthio or alkylsulfonyl) are optionally unsubstituted or independently replaced by 1-4 members selected from halogen, hydroxyl, C 1 -C 3 alkyl, Substituted by at least one of halogenated C 1 -C 3 alkyl, C 1 -C 3 alkoxy or halogenated C 1 -C 3 alkoxy;
  • the R is selected from hydrogen, halogen, hydroxyl, oxo, C1-C6 alkyl acyl, C1-C6 alkyl amido, C1-C6 alkoxy imine, C1-C6 alkyl, C2-C6 alkene C2-C6 alkynyl group, C3-C6 cycloalkyl group, C1-C6 alkoxy group, C6-C10 aryl group, 5-10 membered heteroaryl group containing 1-3 heteroatoms, 1-3 C1-C6 non-aromatic heterocyclic rings of heteroatoms; the above-mentioned groups (including the mentioned alkylacyl, alkylamide, alkoxyimine, alkyl, alkenyl, alkynyl, cycloalkane group, alkoxy group, aryl group, heteroaryl group, non-aromatic heterocycloalkyl group) are optionally unsubstituted or independently selected from halogen, hydroxyl, C
  • the A ring refers to a 5-7 membered saturated or partially unsaturated monocyclic group containing N, m is 1, 2 or 3, n is 1 or 2, preferably, m is 1 and n is 1, Indicates the junction site.
  • R 1 pass Linked to the phenyl group of the compound of formula (1).
  • R 4 or R 5 is a substituent on the heterocycle, which may be connected to a heteroatom or a carbon atom.
  • the heteroatom is selected from N, O or S.
  • R is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, vinyl, ethynyl, cyclopropane group, cyclobutyl, methoxy, ethoxy, n-propoxy, -CH 2 -O-CH 3 , methylsulfonyl or ethylsulfonyl; the above groups are optionally unsubstituted or independent of each other are substituted by C1-C3 alkoxy or halogen.
  • R 4 is selected from H, C1-C3 alkyl, C1-C3 alkoxy substituted C1-C3 alkyl or C1-C3 alkoxy, most preferably methoxymethyl or methoxy.
  • R is selected from hydrogen, F, Cl, Br, hydroxyl, formyl, acetyl, formamido, acetamido, methoxyimino, ethoxyimino, Methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy, ethoxy, phenyl, benzyl, tetrahydrofuryl, hexahydropyranyl, hexahydropyranyl, Hydropyridyl, oxetane, azetidine; the above groups are optionally unsubstituted or independently selected from 1-4 groups selected from F, Cl, Br, hydroxyl, methoxy, and methyl Substituted by at least one of group, tetrahydropyrrole, morpholinyl or -CH 2 -CF 3 .
  • R is selected from H, methyl, ethyl, isopropyl, Cyclopropyl, Fluorocyclopropyl, or
  • ring A refers to a 5-6 membered saturated or unsaturated monocyclic group containing 1, 2 or 3 N ring heteroatoms.
  • the A ring has the following structure:
  • R 5 represents a linking site; said R 5 is as defined in any one of the preceding items of the present application.
  • the A ring has the following structure:
  • R 5 represents a linking site; said R 5 is as defined in any one of the preceding items of the present application.
  • the R 1 is a substituted or unsubstituted pyridine fused heterocyclic ring; more preferably, the R 1 is a substituted or unsubstituted pyridotriazole or a substituted or unsubstituted Pyridimidazole.
  • the R is selected from any one of the following compounds: or
  • R 4 and R 5 are as defined in any one of the foregoing in this application.
  • the R is selected from any one of the following structures:
  • the R is selected from methoxy C1-C3 alkyl or C1-C3 alkoxy; more preferably methoxymethyl or methoxy;
  • the R is selected from H, methyl, ethyl, isopropyl, Cyclopropyl, Fluorocyclopropyl,
  • the compound described in this application has the structure shown in Table 1 below.
  • the application also provides a pharmaceutical mixture, the pharmaceutical mixture comprising two or more compounds selected from the following group: any one of the compounds described in the application or its stereoisomers, tautomers , prodrug, pharmaceutically acceptable salt, amorphous, isotopologue, polymorph or solvate.
  • the present application also provides a pharmaceutical composition, which comprises any one of the compounds described in the present application or its stereoisomers, tautomers, prodrugs, pharmaceutically acceptable salts, amorphous substances, At least one of isotopologues, polymorphs or solvates, and optionally add pharmaceutically acceptable carriers and/or adjuvants.
  • the disease related to GABA A receptor is selected from at least one of the following: pain, Alzheimer's disease, multi-infarct dementia and stroke.
  • the pain is neuropathic pain, inflammatory pain and cancer pain.
  • the pain is selected from: headache, facial pain, neck pain, shoulder pain, back pain, chest pain, abdominal pain, back pain, low back pain, lower limb pain, muscle and bone pain, blood vessel pain, gout, arthritis pain, Visceral Pain, Pain from Infectious Diseases, Pain in Bones, Sickle Cell Anemia, Autoimmune Diseases, Pain Associated with Multiple Sclerosis or Inflammation, Chronic Pain from Injury or Surgery, Nociceptive Pain, Painful Diabetes, Trigeminal Nerve pain, lumbar or cervical radiculopathy, glossopharyngeal neuralgia, autonomic reflex pain, reflex sympathetic dystrophy, nerve root avulsion, cancer, chemical injury, toxin, nutrient deficiency, viral or bacterial infection, or degenerative Pain associated with osteoarthritis.
  • the present application also provides a method for treating or preventing diseases related to GABA A receptors, administering an effective dose of any one of the compounds described in the present application or its stereoisomers, tautomers, pro A body drug, a pharmaceutically acceptable salt, an amorphous substance, a polymorph, a solvate, the pharmaceutical mixture, or the pharmaceutical composition.
  • the application also provides a method for treating or preventing pain, Alzheimer's disease, multi-infarct dementia or stroke, administering to the patient an effective dose of any one of the compound or its stereoisomer, Tautomer, prodrug, pharmaceutically acceptable salt, amorphous, polymorph, solvate, said pharmaceutical mixture, or said pharmaceutical composition.
  • the present application also relates to the preparation method of the compound of formula (1) as described above, comprising:
  • R 3 is ethyl or isopropyl, the experimental operation is the same.
  • the compound of formula (1) and the pharmaceutically acceptable salt thereof of the present application can be prepared by the above-mentioned method.
  • the compound of formula (1) and its intermediate products can be prepared according to the similar method or according to the aforementioned method.
  • Starting materials known in the art may be obtained commercially or may be prepared according to methods known in the art or analogous to known methods.
  • the present application also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a compound as defined above or a pharmaceutically acceptable salt thereof or a prodrug thereof and a pharmaceutically acceptable carrier and/or adjuvant.
  • the present application also includes the above-mentioned compounds or compositions for the preparation of treatment or prevention of ⁇ 2/3-GABA A receptor
  • medicines for diseases of concern especially for the treatment or prophylaxis of the following diseases: pain, epilepsy, anxiety, itching and depression.
  • Pain is preferably treated or prevented.
  • cancer pain refers to the pain that occurs during the development of a malignant tumor.
  • neuroopathic pain is pain provoked or caused by primary damage and dysfunction of the nervous system.
  • inflammatory pain is pain resulting from localized acute inflammation or irritation of nerves by chronic inflammation.
  • treatment also includes prophylactic administration to alleviate or eliminate the condition once it has been established.
  • a "patient” is defined as any warm-blooded animal such as but not limited to a mouse, guinea pig, dog, horse or human, preferably a human.
  • acute pain is defined as pain resulting from noxious stimulation of the skin, body structure, or viscera arising from injury and/or disease, or from abnormal function of muscles or viscera that do not produce actual tissue damage .
  • chronic pain is defined as persistent pain beyond the usual course of an acute illness or a reasonable time for injury to heal, or associated with a chronic pathological process that causes persistent pain, or pain that recurs at intervals of months or years, if Pain that persists after healing has been achieved or persists beyond the usual course of treatment is considered chronic pain.
  • the length of time that elapses in pain depends on the nature of the pain and the course of treatment associated with the pain. Pain is chronic if it exceeds the usual course of treatment.
  • Chronic pain includes, but is not limited to, headache, facial pain, neck pain, shoulder pain, chest pain, abdominal pain, back pain, low back pain, lower extremity pain, musculoskeletal pain, pain associated with somatopathic mental disorder, visceral pain, painful diabetes Neuropathy, vascular pain, gout, arthritic pain, cancer pain, autonomic reflex pain, pain caused by infectious diseases (such as AIDS and herpes zoster), pain caused by autoimmune diseases (rheumatism), acute and chronic Pain due to inflammation, post-surgical pain, and post-burn pain.
  • infectious diseases such as AIDS and herpes zoster
  • autoimmune diseases rheumatism
  • the medicine disclosed in the application can effectively treat chronic pain as defined above, and the medicine disclosed in the application can be used for the treatment of pain sensitivity accompanied by other diseases, including hyperalgesia, allodynia, enhanced pain sensation and pain memory enhancement, and the application will improve treatment of its pain.
  • headache can be divided into primary headache, which includes tension headache, migraine and cluster headache, and secondary headache, which is due to other diseases.
  • primary headache which includes tension headache, migraine and cluster headache
  • secondary headache which is due to other diseases.
  • pain-sensitive tissues of the head and face are lesions or stimulated, various headaches can be caused.
  • These pain-sensitive tissues include the distribution in the scalp, face, mouth and throat, etc., because they are mainly the muscles or blood vessels of the head and contain abundant nerves. Fibers are more sensitive to pain, so when these tissues are damaged it can cause headaches.
  • facial pain includes, but is not limited to, trigeminal neuralgia, atypical facial pain, facial paralysis, and hemifacial spasm.
  • Trigeminal neuralgia is a unique chronic pain disease, also known as painful convulsions, which refers to short-lived, paroxysmal and recurrent electric shock-like severe pain in the distribution area of the trigeminal nerve, Or with ipsilateral muscle spasms. Trigeminal neuralgia is divided into two types: primary and secondary. Primary trigeminal neuralgia refers to no clinical signs of nervous system and no organic disease found in examination; secondary trigeminal neuralgia refers to clinical symptoms. There are neurological signs, and organic lesions, such as tumors and inflammation, are found on examination.
  • atypical facial pain refers to pain caused by multiple etiologies. It manifests as continuous burning pain, without intermission, and has nothing to do with special actions or trigger stimuli. The pain is mostly bilateral, and the pain often exceeds the distribution range of the trigeminal nerve and even involves the neck skin.
  • the etiology can be caused by sinusitis, malignant tumors, jaw and skull base infections and other reasons that stimulate or damage the trigeminal nerve and cause pain.
  • neck pain, back pain, and shoulder pain refer to pain caused by acute and chronic muscle strain, degeneration of bones and joints, trauma, and the like.
  • Common diseases that cause neck, shoulder and upper limb pain include cervical and shoulder myofasciitis, nuchal ligamentitis, cervical spondylosis, frozen shoulder, thoracic outlet syndrome, lateral epicondylitis, etc., or pain caused by autoimmune diseases It is common in diseases such as rheumatoid arthritis, ankylosing spondylitis, and rheumatoid arthritis.
  • Other diseases that may cause neck pain, back pain, and shoulder pain include neck and shoulder tumors, neuritis, arteriovenous diseases, and various diseases. Infection and referred pain caused by thoracic and abdominal visceral lesions.
  • chest, abdomen and back pain refers to pain caused by diseases of the thoracic and abdominal viscera, thoracic and abdominal wall tissues, including but not limited to intercostal neuralgia, intercostal chondritis, angina, abdominal pain (acute abdominal visceral pain) and low back myofascial syndrome.
  • Lumbar and lower limb pain refers to lower back, lumbosacral, sacroiliac, hip, buttock and lower limb pain.
  • Lumbar and lower extremity pain is often not an independent disease, but a common feature of many diseases. The clinical manifestations are diverse, and the etiology is very complex. Most of them are degeneration and injury, including but not limited to lumbar disc herniation, acute lumbar sprain, and sciatica. , osteoporosis, third lumbar transverse process syndrome, piriformis syndrome, knee osteoarthritis, endodynia and heel pain.
  • muscle pain includes, but is not limited to, myofascial pain, trauma-induced pain, and chronic regional pain syndrome Symptoms.
  • ⁇ pain caused by nerve damage complicated by diabetes, which is at least in part due to reduced blood flow and high blood sugar.
  • Some diabetic patients do not develop neuropathy, while others develop the disease at an early stage.
  • Diabetic neuropathy can be divided into mononeuropathy involving one or more lesions and generalized polyneuropathy, which can be diffuse and symmetrical , usually primarily concerned with sensory modality (Merrit's Textbook of Neurology, 9th ed., edited by LPRowland LP).
  • Manifestations of diabetic neuropathy can include autonomic dysfunction leading to dysregulation involving heart, smooth muscle, and glands, causing hypotension, diarrhea, constipation, and impotence.
  • Diabetic neuropathy often develops in stages, early in the nerve endings, autonomic or sensory neuropathy in the feet, cranial neuropathy in the face and around the eyes, intermittent pain and tingling, in subsequent stages, The pain is stronger and more frequent, and finally, painless neuropathy occurs when an area of pain is lost, greatly increasing the risk of severe tissue damage due to the absence of pain as an indicator of injury.
  • visceral pain includes, but is not limited to, the pain of irritable bowel syndrome (IBS), with or without chronic fatigue syndrome (CFS), inflammatory bowel disease (IBD), and interstitial cystitis .
  • IBS irritable bowel syndrome
  • CFS chronic fatigue syndrome
  • IBD inflammatory bowel disease
  • interstitial cystitis interstitial cystitis
  • vascular pain is pain produced by one or more of the following factors.
  • the tissue is improperly perfused.
  • Examples include, but are not limited to, arteriosclerosis obliterans, thromboangiitis obliterans, acute arterial occlusion, embolism, congenital arteriovenous aneurysm, vasospastic disease, Rayaud's disease, cyanosis of extremities, acute venous occlusion, thrombophlebitis, varicose veins and lymphedema.
  • autonomic reflex pain refers to pain resulting from “reflex sympathetic atrophy”.
  • Reflex sympathetic atrophy refers to severe spontaneous pain, hypersensitivity to touch and pain after the body suffers acute and chronic injuries, and may be accompanied by edema and blood flow disorders, followed by symptoms such as skin and musculoskeletal nutritional disorders and atrophy.
  • postoperative pain refers to a complex physiological response of the body to the disease itself and tissue damage caused by surgery, which is manifested as an unpleasant psychological and behavioral experience.
  • arthritic pain includes, but is not limited to, osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthropathy, gout, pseudogout, infectious arthritis, tendonitis, bursa Pain caused by diseases such as inflammation, bone damage and joint soft tissue inflammation.
  • postherpetic neuralgia refers to long-standing severe pain under the skin of the original rash area after the shingles rash has healed.
  • nociceptive pain is pain caused by a tissue damaging process that stimulates nociceptor afferents, or by prolonged excitation of nociceptors. Pain caused by prolonged excitation of nociceptors can be due to persistent noxious stimulation of nociceptors or their sensitization or both, or they can be caused by these factors and be caused by their persistence, various reflex mechanisms, and other factors And extended.
  • the application provides the use of a pharmaceutical compound containing a therapeutically effective amount of ⁇ 2/3-GABA A positive allosteric modulator.
  • ⁇ 2/3-GABA A positive allosteric modulator used in the treatment of the present application can be administered in the form of a raw compound, it is preferred that the active ingredient, optionally in the form of a physiologically acceptable salt, be combined with one or Various additives, excipients, carriers, buffers, diluents and/or other conventional pharmaceutical adjuvants are mixed together to form a pharmaceutical composition.
  • the application provides a pharmaceutical composition containing an ⁇ 2/3-GABA A positive allosteric modulator, wherein the ⁇ 2/3-GABA A positive allosteric modulator is combined with one or more pharmaceutically acceptable carrier, and optionally in admixture with other therapeutic and/or prophylactic ingredients known or used in the art.
  • the carrier must be "acceptable”, ie, compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
  • the compounds used in the present application can be formulated together with conventional additives, or diluents, in the form of pharmaceutical compositions and unit doses thereof.
  • forms such as these include solids (especially in the form of tablets, filled capsules, powders and pills), and liquids (especially aqueous or non-aqueous solutions, suspensions, emulsions, elixirs), and capsules filled with the above forms, all oral Forms for administration, suppositories for rectal administration, and sterile injectable solutions for parenteral administration.
  • Pharmaceutical compositions such as these and unit dosage forms thereof may comprise conventional ingredients in conventional proportions, with or without additional active compounds or ingredients, and such unit dosage forms may contain any suitable effective dose commensurate with the desired daily application dosage range. amount of active ingredient.
  • the compounds used in this application can be administered in a variety of oral and parenteral dosage forms. It will be apparent to those skilled in the art that the following dosage forms may contain, as an active ingredient, a compound of the present application or a pharmaceutically acceptable salt thereof.
  • pharmaceutically acceptable carriers can be solid or liquid.
  • Solid form preparations include powders, tablets, doses, capsules, cachets, suppositories, and dispersible granules.
  • a solid carrier can be one or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.
  • the carrier is a finely divided solid which is in admixture with the finely divided active ingredient.
  • the active ingredient is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
  • Powders and tablets preferably contain from 5% or 10% to about 70% active compound.
  • Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting waxes , cocoa butter, etc.
  • the term "preparation" includes the active compound formulated with encapsulating material as carrier, which provides a capsule in which the active component, with or without carriers, is surrounded by a carrier, which is thus in association with it.
  • the formulations include cachets and lozenges. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid forms suitable for oral administration.
  • a low-melting wax such as fatty acid glycerides or a paste of cocoa butter is first melted and the active ingredient is dispersed uniformly therein by stirring. This molten homogeneous mixture is then poured into appropriately sized molds, allowed to cool and thereby solidify.
  • compositions suitable for vaginal administration may be in the form of pessaries, tampons, creams, gels, pastes, foams or sprays and contain, in addition to the active ingredient, suitable carriers known in the art. .
  • Liquid preparations include solutions, suspensions and emulsions, for example, aqueous solutions or water-propylene glycol solutions.
  • liquid preparations for parenteral injection can be formulated as water-polyethylene glycol solutions.
  • the compounds for use in this application may thus be formulated for parenteral administration (e.g., injection, such as bolus injection or continuous infusion), and may be presented in unit dosage form in ampoules, prepregs, with an added preservative. Filled syringes, small-volume infusion bags, or multi-dose containers.
  • the compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulation ingredients such as suspending, stabilizing and/or dispersing agents.
  • the active ingredient may be in powder form, aseptically isolated from sterile solid or lyophilized from solution, for reconstitution with a suitable vehicle, eg sterile, pyrogen-free water, before use.
  • Aqueous solutions suitable for oral administration can be prepared by dissolving the active ingredient in water and adding required colorants, flavours, stabilizing and thickening agents.
  • Aqueous suspensions suitable for oral administration can be prepared by dispersing the finely divided active ingredient in viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other known suspensions. prepared in the water of the agent.
  • viscous material such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other known suspensions. prepared in the water of the agent.
  • liquid preparations designed to be converted, shortly before use, to liquid formulations for oral administration.
  • liquid preparations include solutions, suspensions and emulsions.
  • Such preparations may contain, in addition to the active ingredient, coloring agents, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like.
  • the compounds of the present application may be formulated as ointments, creams or lotions or as a transdermal patch.
  • Ointments and creams for example, may be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents.
  • Lotions may be formulated with an aqueous or oily base, and generally also contain one or more emulsifying, stabilizing, dispersing, suspending, thickening or coloring agents.
  • compositions suitable for topical administration in the oral cavity include lozenges containing the active ingredient in a flavored base, usually sucrose and acacia or tragacanth; inert bases such as gelatin and glycerin or sucrose and acacia pastilles containing the active ingredient; and mouthwashes containing the active ingredient in a suitable liquid carrier.
  • solutions or suspensions can be applied directly to the nasal cavity by conventional means, eg, with a dropper, pipette or spray.
  • the composition may be in single or multiple dose form.
  • Respiratory administration can also be achieved by aerosol, wherein the active ingredient is contained in a pressurized pack with a suitable propellant including chlorofluorocarbons (CFCs) such as dichlorodifluoromethane, trichlorofluoromethane, Methane or dichlorotetrafluoroethane, carbon dioxide or other suitable gases.
  • a suitable propellant including chlorofluorocarbons (CFCs) such as dichlorodifluoromethane, trichlorofluoromethane, Methane or dichlorotetrafluoroethane, carbon dioxide or other suitable gases.
  • Aerosol formulations may also suitably contain a surfactant, such as lecithin.
  • the dosage of the drug can be controlled by a dosage valve.
  • the active ingredient may be in the form of a dry powder, for example a powder mix of the compound with a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidone (PVP).
  • a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidone (PVP).
  • the powder carrier conveniently forms a gel in the nasal cavity.
  • Powder compositions may be presented in unit dosage form, for example, in capsules or cartridges, eg gelatin capsules or cartridges, or in blister packs from which the powder can be administered via an inhaler.
  • compositions for respiratory administration typically the compound has a small particle size, eg, on the order of 5 microns or less.
  • particle sizes can be obtained by methods known in the art, for example by micronization.
  • compositions suitable for sustained release of the active ingredient may be used, if desired.
  • compositions are optionally presented in unit dosage form.
  • the preparation is subdivided into unit doses of appropriate quantities of the active ingredient.
  • the unit dosage form can be a packaged preparation, where the hermetically sealed package contains discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules.
  • the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of such in packaged form.
  • Tablets or capsules for oral administration and liquids for intravenous administration and continuous infusion are preferred compositions.
  • the amount of the active ingredient in the unit dosage preparation can vary according to the specific application and the potency of the active ingredient, and can be adjusted from 0.01 mg to about 0.1 g.
  • the drug in pharmaceutical use, may be administered in capsules of 0.01 to about 100 mg three times daily, and the composition may also contain other compatible therapeutic agents if desired.
  • the compounds used in the present application are used in a starting dose of 0.001 mg/kg to 10 mg/kg body weight per day.
  • these dosages may vary according to the needs of the patient, the severity of the condition being treated, and the compound employed, and generally, treatment is initiated with smaller dosages less than the optimum dosage of the compound and thereafter, the dosage is increased by small amounts to reach the optimal dosage.
  • the total daily dosage may be subdivided into divided doses throughout the day if necessary.
  • the pharmaceutical composition of the present application can also be used in combination with other drugs for treating pain, epilepsy, anxiety and depression, including but not limited to morphine, gabapentin and the like. Therefore, the application provides a drug for the treatment of pain, epilepsy, anxiety and depression, which is not only effective, but also has no obvious side effects. Another purpose of the application is to provide a drug for special patient populations, such as It is a drug with high safety for the elderly, patients with liver or kidney function decline, or patients with cardiovascular diseases.
  • the present application also provides a method for treating or preventing a disease, comprising administering an effective dose of the above-mentioned compound or composition to a patient.
  • the present application also provides a method for treating or preventing diseases related to GABA A receptors, comprising administering an effective dose of the above-mentioned compound or the above-mentioned composition to the patient.
  • the present application also provides a use of the above-mentioned compound or composition in the preparation of medicines for treating or preventing the following diseases: pain, Alzheimer's disease, multi-infarct dementia and stroke.
  • Said pain is neuropathic pain, inflammatory pain and cancer pain.
  • the pain is selected from the group consisting of: headache, facial pain, neck pain, shoulder pain, back pain, chest pain, abdominal pain, back pain, lumbago, lower limb pain, musculoskeletal pain, vascular pain, gout, arthritis pain, Visceral Pain, Pain from Infectious Diseases, Pain in Bones, Sickle Cell Anemia, Autoimmune Diseases, Pain Associated with Multiple Sclerosis or Inflammation, Chronic Pain from Injury or Surgery, Nociceptive Pain, Painful Diabetes, Trigeminal Nerve pain, lumbar or cervical radiculopathy, glossopharyngeal neuralgia, autonomic reflex pain, reflex sympathetic dystrophy, nerve root avulsion, cancer, chemical injury, toxin, nutrient deficiency, viral or bacterial infection, or degenerative Pain associated with osteoarthritis.
  • the present application also provides a method for treating or preventing pain, Alzheimer's disease, multi-infarct dementia or stroke, by administering an effective dose of the compound as described above or the composition as described above to the patient.
  • the compound of the present application and its pharmaceutically acceptable salt or its prodrug have important pharmacological properties, and are positive allosteric modulators of ⁇ 2/3-GABA A receptor.
  • the compound of the present application has excellent affinity activity and positive regulation activity on ⁇ 2/3-GABA A receptor, and has good genotoxicity safety and druggability. Therefore, the compounds of the present application and their pharmaceutically acceptable salts or prodrugs thereof can be used alone or in combination with other drugs for the treatment or prevention of diseases related to ⁇ 2/3-GABA A.
  • the ratio of solvents used in the purification steps is volume ratio.
  • reaction solution was diluted with ethyl acetate (50 mL), washed with saturated aqueous sodium chloride (50 mL ⁇ 3), the organic phase was dried, concentrated, and purified by preparative HPLC to obtain the title product (30 mg, 17%) as a white solid.
  • 2-Fluoro-3-methoxypyridine (5g, 39.3mmol) was dissolved in anhydrous tetrahydrofuran (150mL), and the reaction solution was cooled to -60°C under nitrogen protection, and n-butyllithium (2.5 M) (23.6mL, 59mmol), control the temperature at -60 ⁇ -65°C for 10min, and stir at -60°C for 1h.
  • Example 2 The experimental operation is the same as in Example 2, with 7-iodo-8-methoxy-3-(3-methoxycyclobutyl)-[1,2,4]triazolo[4,3-a]pyridine ( 100.0mg, 0.278mmol), (2-fluoro-5-(7-isopropyl-7H-imidazo[4,5-c]pyridazin-4-yl)phenyl)boronic acid (100.3mg, 0.334mmol) As starting material, a mixture of the two titles was obtained as a white solid. Further chiral resolution was carried out to obtain Example 3 and Example 4.
  • Example 3 The first peak, 14 mg, white solid, yield 10%.
  • Example 4 The second peak, 38mg, white solid, yield 28%.
  • the experimental operation is the same as in Example 2, with 7-iodo-8-methoxy-3-(2-methyltetrahydro-2H-pyran-4-yl)-[1,2,4]triazolo[4 ,3-a]pyridine (100mg, 0.27mmol), (2-fluoro-5-(7-ethyl-7H-imidazo[4,5-c]pyridazin-4-yl)phenyl)boronic acid (77mg , 0.27mmol) as starting material to obtain the title compound (35mg, white solid) with a yield of 27.0%.
  • Methyl 6-bromo-7-methoxyimidazo[1,2-a]pyridine-2-carboxylate 400 mg, 1.4 mmol
  • (2M) aqueous sodium hydroxide solution 2 mL
  • 6-Bromo-7-methoxyimidazo[1,2-a]pyridine-2-carboxylic acid 300mg, 1.1mmol
  • 1-hydroxybenzotriazole (234mg, 1.1mmol)
  • 1-(3- Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride 95mg, 0.5mmol
  • N,N-diisopropylethylamine 425mg, 3.3mmol
  • pyrrolidine 213mg, 3.3mmol
  • Example 2 The experimental operation is the same as in Example 2, with 4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)- 7-Isopropyl-7H-imidazo[4,5-c]pyridazine (45 mg, 0.11 mmol) and 7-iodo-8-methoxy-3-(tetrahydro-2H-pyran-4-yl )-[1,2,4]triazolo[4,3-a]pyridine (50mg, 0.14mmol) was used as starting material to obtain the title compound (40mg, 74.5%).
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl ) phenyl)-7H-imidazo[4,5-c]pyridazine (100mg, 0.3mmol), 4-chloro-7-ethyl-7H-imidazo[4,5-c]pyridazine (5mg, 0.3 mmol) as starting material to give the title compound (17 mg, 16%) as a white solid.
  • Example 2 The experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)-7H-imidazo[4,5-c]pyridazine (42mg, 0.115mmol), 3-(4-fluoro-1-methylpiperidin-4-yl)-7-iodo- Starting from 8-methoxy-[1,2,4]triazolo[4,3-a]pyridine (35 mg, 0.115 mmol), the title compound was obtained as a yellow solid (13 mg, 27%).
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)-7H-imidazo[4,5-c]pyridazine (62mg, 0.17mmol), 7-iodo-8-methoxy-3-(1-(2,2,2-tri Fluoroethyl)azetidin-3-yl)-[1,2,4]triazolo[4,3-a]pyridine (70mg, 0.17mmol) as raw material, the title compound was obtained as a pale yellow solid (22mg ,25%).
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)-7H-imidazo[4,5-c]pyridazine (54mg, 0.15mmol), 7-iodo-8-methoxy-3-(4-methyltetrahydro-2H-pyridazine Using pyran-4-yl)-[1,2,4]triazolo[4,3-a]pyridine (55mg, 0.15mmol) as starting material, the title compound was obtained as a yellow solid (19mg, 27%).
  • the experimental operation is the same as in Example 2, with 6-bromo-7-methoxy-2-methylimidazo[1,2-a]pyridine (24.2mg, 0.1mmo) and 7-isopropyl-4-(4 -Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-7H-imidazo[4,5-c]pyridine Using oxazine (40 mg, 0.1 mmol) as starting material, the title compound (17 mg, 42%) was obtained.
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)-7H-imidazo[4,5-c]pyridazine (56mg, 0.151mmol), 7-iodo-8-methoxy-3-(oxetan-3-yl) -[1,2,4]triazolo[4,3-a]pyridine (50mg, 0.151mmol) was used as starting material to obtain the title compound as a yellow solid (26mg, 39%).
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)-7H-imidazo[4,5-c]pyridazine (58mg, 0.158mmol), 7-iodo-8-methoxy-3-isopropyl-[1,2,4] Triazolo[4,3-a]pyridine (50mg, 0.158 mmol) as starting material to obtain the title compound as a yellow solid (17 mg, 25%).
  • the experimental operation is the same as in Example 2, with 6-bromo-2-ethyl-7-methoxyimidazo[1,2-a]pyridine (50mg, 0.20mmol), 7-ethyl-4-(4-fluoro -3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-7H-imidazo[4,5-c]pyridazine ( 74mg, 0.20mmol) as starting material to obtain the title compound (10mg, 12%) as a brown solid.
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)-7H-imidazo[4,5-c]pyridazine (60mg, 0.165mmol), 7-iodo-8-methoxy-3-ethyl-[1,2,4]tri Azolo[4,3-a]pyridine (50mg, 0.165mmol) was used as starting material to give the title compound as a yellow solid (23mg, 33%).
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)phenyl)-7H-imidazo[4,5-c]pyridazine (67mg, 0.182mmol), 7-iodo-8-methoxy-3-methyl-[1,2,4]tri Azolo[4,3-a]pyridine (50mg, 0.182mmol) was used as starting material to give the title compound as a yellow solid (27mg, 37%).
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )phenyl)-7H-imidazo[4,5-c]pyridazine (60mg, 0.17mmol), 7-iodo-8-methoxy-3-(tetrahydrofuran-3-yl)-[1,2, 4] Starting from triazolo[4,3-a]pyridine (50mg, 0.17mmol), the title compound (47mg, 60%) was obtained as a yellow solid.
  • the experimental operation is the same as in Example 2, with 6-bromo-2-cyclopropyl-7-methoxyimidazo[1,2-a]pyridine (150mg, 0.56mmol), 7-ethyl-4-(4- Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-7H-imidazo[4,5-c]pyridazine (249mg, 0.67mmol) as starting material to obtain the title compound (83mg, 34%).
  • the experimental operation is the same as in Example 2, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )phenyl)-7H-imidazo[4,5-c]pyridazine (54mg, 0.15mmol), 5-bromo-6-methoxy-2-methylimidazo[1,2-a]pyridine ( 35 mg, 0.15 mmol) as starting material to give a brown solid (25 mg, 41% yield).
  • the experimental operation is the same as in Example 2, with 7-bromo-6-methoxy-[1,2,4]triazolo[4,3-a]pyridine (30mg, 0.13mmol), 7-ethyl-4- (4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-7H-imidazo[4,5-c ] Pyridazine (48 mg, 0.13 mmol) was used as starting material to obtain the title compound (3 mg, 6%) as a yellow solid.
  • the experimental operation is the same as in Example 2, with 6-bromo-7-methoxyimidazo[1,2-a]pyridine-3-methanol (40mg, 0.16mmol), 7-ethyl-4-(4-fluoro- 3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-7H-imidazo[4,5-c]pyridazine (59mg , 0.16mmol) as starting material to obtain the title compound (6mg, 8%) as a brown solid.
  • the experimental operation is the same as in Example 19, with 7-ethyl-4-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolin-2-yl ) phenyl)-7H-imidazo[4,5-c]pyridazine (74mg, 0.2mmol), 3-bromo-2-methoxy-1,8-naphthyridine (40mg, 0.17mmol) as raw materials, The title compound (38 mg, 48%) was obtained.
  • the experimental operation is the same as in Example 19, with 6-bromo-7-methoxyimidazo[1,2-a]pyrimidine (57mg, 0.251mmol), 7-ethyl-4-(4-fluoro-3-(4 ,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-7H-imidazo[4,5-c]pyridazine (92mg, 0.251mmol) As starting material, the title compound (50mg, 52%) was obtained.
  • the experimental operation is the same as in Example 2, with 6-bromo-7-methoxy-2-methylimidazo[1,2-a]pyridine (30mg, 0.12mmol), 7-ethyl-4-(4-fluoro -3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-7H-imidazo[4,5-c]pyridazine ( 44mg, 0.12mmol) as starting material to obtain the title compound (13mg, 27%) as a white solid.
  • Example 55 1 H NMR (400 MHz, DMSO-d6) ppm 1.50-1.57 (m, 3H) 2.26-2.32 (m, 3H) 3.80-3.85 (m, 3H) 3.89-3.97 (m, 3H) 4.47-4.54 (m,2H)7.12-7.17(m,1H)7.52-7.58(m,1H)8.44-8.59(m,3H)8.65-8.71(m,1H)8.83-8.89(m,1H)9.52-9.60(m ,1H).
  • LC- MS: m/z[M+H] + 460
  • Methylmagnesium chloride (620 mg, 8.35 mmol) and methyl 6-bromo-7-methoxyimidazo[1,2-a]pyridine-2-carboxylate (500 mg, 1.67 mmol) were added to tetrahydrofuran (20 mL) , and stirred at room temperature for 1 hour.
  • the reaction solution was adjusted to be acidic with aqueous hydrochloric acid (1N), concentrated, then adjusted to be alkaline with saturated aqueous sodium bicarbonate, extracted with dichloromethane (50mL ⁇ 3), and the organic phase was washed with saturated brine (50mL).
  • the experimental operation is the same as in Example 1, with 7-bromo-3-methyl-8-(methoxymethyl)-[1,2,4]triazolo[4,3-a]pyridine (143mg, 0.56mmol ), (5-(7-ethyl-7H-imidazo[4,5-c]pyridazin-4-yl)-2-fluorophenyl)boronic acid (159mg, 0.56mmol) as starting material, the title compound was obtained as brown Solid (82mg, 35%).
  • the ⁇ , ⁇ and ⁇ subunits are essential to form a fully functional GABA A receptor.
  • the present application constructs and selects stably expressing human ⁇ 1- HEK293 cells for GABA A receptors ( ⁇ 1-GABA AR ) and ⁇ 2-GABA A receptors ( ⁇ 2-GABA AR ).
  • the ⁇ 1-GABA A R-HEK293 cell model simultaneously expresses ⁇ 1 subunit (for protein sequence, see GenBank accession number: NM_000806.5), ⁇ 3 subunit (for protein sequence, see GenBank accession number: NM_000814.5) and ⁇ 2 subunit (for protein sequence, see GenBank accession number: NM_000816.3).
  • the ⁇ 2-GABA A R-HEK293 cell model simultaneously expresses ⁇ 2 subunit (for protein sequence, see GenBank accession number: NM_000807.4), ⁇ 3 subunit (for protein sequence, see GenBank accession number: NM_000814.5) and ⁇ 2 subunit (for protein sequence, see GenBank accession number: NM_000816.3).
  • the above cell lines were subcultured.
  • the ⁇ 2-GABA A R-HEK293 cells were expanded for the benzodiazepine site (BZD) affinity test of the compound on the GABA A receptor.
  • BZD benzodiazepine site
  • ⁇ 1-GABA A R-HEK293 cells and ⁇ 2-GABA A R-HEK293 cells were partly suspended in the process of subculture and spread on slides pretreated with Poly-D-Lysine (polylysine) for electrophysiological tests ( The method refers to paragraph 0586 of patent CN 107344936A).
  • the affinity of the compound to the ⁇ 2-GABA A receptor was determined by competing for the binding of 3 H-flunitrazepam (isotope 3 H-labeled flunitrazepam) to the membrane protein BZD site of R-HEK293 cells stably expressing human ⁇ 2-GABA A.
  • Tris buffer 50 mM Tris-HCl, 10 mM MgCl 2 , 0.5 mM EDTA, 10% glycerol.
  • Radioligand competition binding assay This assay was carried out in a 200 ⁇ L system (96-well plate) containing 100 ⁇ L of cell membranes. The concentration of 3 H-flunitrazepam was 1 nM, and the concentration of the test compound was in the range of 1x 10 -5 -10 -6 M. Take flumazenil (flumazenil) as the control. 1 ⁇ L of 2 mM flumazenil (final concentration 10 ⁇ M) was added to low signal control wells (Low control, LC), and 1 ⁇ L of dimethyl sulfoxide was added to high signal control wells (High control, HC). The final target membrane protein concentration was 5 ⁇ g/well.
  • All test compound sample stock solution is 10mM dimethyl sulfoxide solution.
  • the working concentration of the sample was to dilute all the samples to 0.2 mM with dimethyl sulfoxide, and then perform 4-fold serial dilution, with a total of 8 concentration gradients.
  • PEI polyethyleneimine
  • the cells were collected on the GF/C filter plate with a cell harvester, and washed 4 times with plate washing buffer (50 mM Tris-HCl, pH 7.4, stored at 4°C). After drying in an oven at 50°C for 1 hour, seal the bottom of the dried GF/C filter plate, detect the residual radioactivity of the filter membrane by liquid scintillation counting method, add 50 ⁇ L of scintillation liquid to each well, and seal it.
  • plate washing buffer 50 mM Tris-HCl, pH 7.4, stored at 4°C.
  • Table 1 shows representative detection results obtained by the method for determining the binding affinity of the compound to the BZD site of the cell membrane protein expressing human ⁇ 2-GABA A R-HEK293.
  • the positive regulatory activity of the drug to be tested on the ⁇ 1-GABA A receptor and the ⁇ 2-GABA A receptor is detected by an electrophysiological method.
  • the specific method is as follows:
  • Compound concentration setting the final concentration of the compound used in compound screening is 100nM.
  • concentration of GABA is 0.05 ⁇ 0.07 ⁇ M (about EC 2 ⁇ 4 );
  • concentration of GABA is 0.10 ⁇ 0.11 ⁇ M (about EC 7 ⁇ 8 ).
  • the whole-cell patch clamp technique was used for the electrophysiological test, which can refer to the method reported in the literature (Nickolls, SA, et al. British Journal of Pharmacology, 2018, 175:708-725).
  • the composition of extracellular solution (ECS) for electrophysiology is as follows: 150mM NaCl, 5mM KCl, 2.5mM CaCl 2 , 1mM MgCl 2 , 10mM HEPES and 10mM glucose (pH7.4); the formula of electrode inner solution (ICS) for electrophysiology is as follows: 140 mM CsCl, 11 mM EGTA, 10 mM HEPES, 2 mM CaCl 2 , 1 mM MgCl 2 , 4 mM MgATP, 2 mM TEA (pH 7.3).
  • the GABA ( ⁇ -aminobutyric acid) powder was prepared into a mother solution with pure water, and then diluted in ECS; the compound was first prepared with dimethyl sulfoxide into a 4mM mother solution, and then gradually diluted into GABA-ECS of the corresponding concentration. Solutions were prepared freshly before electrophysiological testing.
  • Electrophysiological signals were collected using EPC 10 amplifier and PatchMaster software (HEKA) or Axon700B amplifier and Clampex software (AXON).
  • the recording electrodes are drawn from borosilicate glass, and the electrode resistance is 4-6 M ⁇ .
  • ALA-VC-8PG TM system was used for extracellular drug delivery. Select a single cell that grows independently, and the glass electrode forms a good seal with the cell and then ruptures the membrane to form a whole-cell model.
  • the cell membrane potential was clamped at -60mV and recorded in Gap-free mode.
  • extracellular fluid was first applied outside the cells for about 20 seconds. After the baseline (I prebaseline ) stabilized, the extracellular fluid was switched to GABA-ECS.
  • a GABA-induced current (I gaba ) can be detected.
  • the extracellular fluid is switched to the mixed solution of the compound and GABA-ECS until the current (I treatment ) jointly induced by the compound and GABA can be detected.
  • switch the solution to extracellular fluid and record for 20-40 seconds to terminate the test. Only cells with a stable baseline, an absolute value of the control current (I gaba -I prebaseline ) greater than 40pA, and a relatively stable current within 10-20 seconds will be used for compound testing.
  • the value is negative, it means that the modulation effect of the test compound on the GABA receptor is inverse allosteric modulation. If the value is positive, it means that the modulation effect of the test compound on the GABA receptor is a positive allosteric modulation.
  • Table 1 shows the positive allosteric regulation activity test results of the compounds on ⁇ 1-GABA A receptor and ⁇ 2-GABA A receptor.
  • the experimental design is based on the experimental purpose and follows the following experimental design guidelines:
  • CHO-WBL cells Chinese hamster ovary cells
  • the karyotype of this cell line contains 21 chromosomes, and the growth cycle is 12 to 13 hours.
  • CHO-WBL cells were originally derived from Dr. S. Wolff of the University of California, San Francisco, USA, cloned in the laboratory of Dr. A. Bloom of Columbia University in New York, and then cloned again by Dr. S. Galloway of Litton Bionetics Laboratory in Maryland .
  • Cell stocks were stored in liquid nitrogen. Each batch of cell stock solution has its karyotype identified and has been tested to prove that it is not contaminated by mycoplasma.
  • Metabolic activation system The in vitro metabolic activation system (Maron and Ames, 1983) consists of rat liver microsomal homogenate (S9 for short) induced by ⁇ -naphthoflavone and phenobarbital and a NADPH production system (i.e. NADP disodium plus isosome trisodium citrate). S9 was purchased from Qishi Biotechnology Co., Ltd. and stored in a -75°C refrigerator before use. Preparation method of S9 homogenate: ⁇ -naphthoflavone and phenobarbital were intraperitoneally injected into male Sprague Dawley rats once, and the materials were prepared.
  • Preparation of the S9 metabolic activation system Dissolve NADP disodium salt and isocitrate trisodium salt in serum-free medium to prepare a core solution, and filter it with a 0.22 ⁇ m filter head to sterilize. Before use, the S9 homogenate was melted, and the components of the S9 metabolic activation system were mixed according to the ratios shown in Table 2 below.
  • Dose-finding experiment In the dose-finding experiment, 8 dosing concentrations were set up in each dosing treatment series. Set up 1 well of cells for each concentration of the test substance.
  • ICH guideline S2(R1) when not limited by the solubility of the test substance in the solvent/medium or the cytotoxicity of the test substance, the recommended maximum concentration is 1mM or 0.5mg/mL, whichever is lower By. However, if limited by solubility, the highest concentration tested was the lowest concentration at which a small amount of precipitate was visible in the medium under a microscope. To ensure that the upper limit of solubility in the medium is reached, up to 4 concentrations with visible precipitation can be tested.
  • Dosing treatment In the 3-hour dosing series of S9 metabolic activation, suck off the medium in each well, and add serum-free McCoy's 5A/S9 containing appropriate concentration of test substance/control substance according to the ICH S2(R1) guidelines mixed media.
  • McCoy's 5A complete medium containing cytochalasin B (final concentration in the medium was about 3 ⁇ g/mL) was added to each well, followed by culturing for 21 hours until harvesting.
  • the treatment is similar to the non-metabolically activated 3-hour dosing series, except that the cells will be mixed with the test substance/control substance in a culture medium containing cytochalasin B (final concentration in culture medium is approx. 3 ⁇ g/mL) in the whole medium 24 hours ( ⁇ 30 minutes).
  • Slide preparation when harvesting, discard the culture medium in the eight-well slide, blot dry with absorbent paper, add 75mM KCl hypotonic solution to each well, and then replace the cells with fixative solution (anhydrous methanol: glacial acetic acid) twice consecutively. , 3:1v/v) fixed for 5 minutes and air-dried. Stain with acridine orange and air dry in the dark.
  • fixative solution anhydrous methanol: glacial acetic acid
  • Counting Cytotoxicity By examining the apparent staining under a low power microscope, assess the likelihood that all wells of each slide will be analyzable (sufficient number of cells). Under a fluorescence microscope, at least 500 cells were analyzed per well, and the numbers of mononucleated, binucleated and multinucleated cells contained were counted to calculate the cytokinesis block proliferation index (CBPI).
  • CBPI cytokinesis block proliferation index
  • CBPI The cytotoxicity of the test substances was evaluated by CBPI.
  • the calculation formula of CBPI is as follows:
  • T test substance treatment culture group
  • C vehicle control culture group
  • Micronucleus main experiment In the micronucleus main experiment, CHO-WBL cells were exposed to 3-8 concentrations of the test substance, and a positive control group (cyclophosphamide monohydrate) and a vehicle control group were set up with the same treatment as the solvent group. (dimethyl sulfoxide), each dose group is equipped with 2 wells of cells. In the non-activated test system, the administration time was 3 and 24 hours, and in the S9 activated test system, the test substance exposure time was 3 hours. The upper limit of the test substance test depends on its solubility in the medium, but in general will not exceed the maximum concentration of 1mM or 0.5mg/mL, whichever is lower.
  • the cytotoxicity of the test substance dose group should not exceed 50% compared with the corresponding vehicle control group. If the highest concentration is not limited by cytotoxicity, in order to reach the upper limit of solubility in the medium, multiple doses with visible precipitate can be tested, and a small amount of identifiable precipitate can be seen in the medium of the highest dose group.
  • Dosage selection for micronucleus analysis The choice of dose for micronucleus analysis of CHO-WBL cells will depend on the cytotoxicity of the test substance. The highest dose chosen for evaluation will induce no more than 50% more cytotoxicity. At least two further lower dose groups (in the moderate cytotoxic to non-cytotoxic range, as applicable) were analyzed. If the test substance has no cytotoxicity, select the dose at which precipitation can be observed under the microscope at the end of administration, or the maximum concentration to be tested (1mM or 0.5mg/mL, whichever is lower) is the highest concentration for micronucleus analysis. concentration.
  • Counting the frequency of micronuclei All slides selected for micronucleus analysis were marked with randomly generated blind codes to reduce the subjectivity of slide reading. The blind codes were made by personnel who did not participate in the slide reading. Blindly labeled slide labels contain the following information: experiment number, treatment series, and random number.
  • Binuclear cells should meet the following criteria to be counted:
  • a. Has a complete cell membrane.
  • b. The two sub-nuclei are equal in size.
  • microkernel The criteria for judging the microkernel are as follows:
  • Micronuclei have similar fluorescence intensity as the main nuclei. b. Micronuclei should be free in the cytoplasm. c. The micronuclei have smooth edges, and the diameter is less than or equal to one-third of the diameter of the main nucleus.
  • Assay Validity Criteria An acceptable genotoxicity test must meet the following criteria: The frequency of vehicle/negative control MNs must be comparable to historical negative control data. There should be at least three concentrations that can be analyzed. The proportion of micronucleated cells in the positive control should have a statistically significant increase (p ⁇ 0.05) compared to the parallel vehicle/negative control group.
  • test substance If the test substance satisfies the following criteria at the same time, it can be considered as positive: the frequency of micronucleus cells in one or more dose groups is significantly increased compared with the parallel vehicle control group. The increase in the frequency of micronuclei has a dose-effect relationship. Micronucleus frequencies outside the range of laboratory negative historical data were observed in one or more dose groups.
  • test substance only partially meets the above three points, it should be judged scientifically. Signs of concentration-related effects were considered useful, but not necessary in evaluating a positive result. Biological relevance will be considered, such as consistency of responses within and between concentrations and, where applicable, between experiments, or effects that occur only at high or extremely cytotoxic concentrations.
  • the compounds of the examples of the present application have better selectivity, and make the electrophysiological activity of the ⁇ 1-GABA A receptor less than that of CVL-865 while maintaining an appropriate ⁇ 2-GABA A receptor affinity and electrophysiological activity 70% of the drug, so that while maintaining the efficacy of the drug in the body, there are fewer side effects.

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Abstract

提供了一种咪唑并哒嗪类衍生物、其制备方法、药物组合物和用途。具体提供了一种式(1)所示化合物、其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、同位素体、多晶型物或溶剂合物，含有该化合物的药物组合物以及所述化合物作为GABA A受体调节剂的用途。

Description

咪唑并哒嗪类衍生物、其制备方法、药物组合物和用途
相关申请的交叉引用
本申请要求在2022年2月25日提交中国专利局、申请号为202210178772.9、发明名称为“咪唑并哒嗪类衍生物、其制备方法、药物组合物和用途”的中国专利申请的优先权，其全部内容通过引用的方式并入本文中。
技术领域
本申请涉及对GABAA受体具有调节功能的咪唑并哒嗪类衍生物、其制备方法、药物组合物和它们作为药物的应用。
背景技术
γ-氨基丁酸(GABA)是哺乳动物中枢神经系统中重要的抑制性神经递质。调节GABA能神经传导的物质，被广泛地用于治疗各种病症，诸如癫痫症、焦虑症和抑郁症。自然界中，有两类GABA受体，一类是GABAA受体(GABAAR)，该类受体为配体门控离子通道超家族的成员，另一类是GABAB受体(GABABR)，该类受体是为G蛋白偶联受体超家族的成员。哺乳动物中的GABAA受体亚基已发现的有α1-6、β1-4、γ1-3、δ、ε、θ和ρ1-2等亚基，其中α亚基、β亚基和γ亚基对形成一个完整的功能型GABAA受体是必不可少的，而α亚基对苯二氮卓类化合物与GABAA受体的结合是至关重要的。
在别构结合位置上结合的药物可为增加受体活性的阳性别构调节剂(或正向别构调节剂)、降低受体活性性的阴性别构调节剂(或反向别构调节剂)或不改变受体活性的中性别构调节剂(该指与别构结合位置结合但不调节受体活性的化合物)。最近的证据示意包含α2或α3亚基的GABAA受体(在本文称为α2/3-GABAA受体)可能涉及某些疼痛状态，且这些受体的阳性别构调节剂可能为有效的镇痛剂(Mirza,N.R.和Munro,G.,Drug News and Perspectives,2010,23(6),351-360)。
国际专利申请案PCT/GB01/04948(公告为WO2002/038568)和PCT/GB02/03114(公告为WO2003/008418)描述7-苯基咪唑并[1,2-b][1,2,4]三嗪衍生物，其对α2、α3和/或α5亚基具有亲和性。国际专利申请案PCT/US99/14935(公告为WO2000/001697)尤其公开了4-苯基-7H-咪唑并[4,5-c]哒嗪衍生物，其为促肾上腺皮质素释放因子拮抗剂。国际专利申请案PCT/IB2013/060631(公告为WO2014/091368)和PCT/IB2015/054200(公告号为WO2015/189744)和Robert M.Owen的文章(Robert M.Owen的J.Med.Chem.2019,62,5773-5796)描述4-(联苯-3-基)-7H-咪唑并[4,5-c]哒嗪衍生物与α2/3-GABAA受体具有相互作用，用于治疗包括疼痛等多种疾病。该类化合物还被用于瘙痒症(国际专利申请案PCT/US2019/033598，公告号为WO2019/26820A1)以及癫痫(Duveau,V.的CNS Neurosci Ther.2019.25(2):p.255-260.)的治疗。
主流观点认为对含有α1亚基的GABAA受体的调节活性，是目前GABAA调节剂(诸如苯并二氮杂卓类)的副作用(诸如镇静、成瘾性、嗜睡、健忘)主要来源(Uwe Rudolph和Frederic Knoflach的Nature Reviews:Drug Discovery,2011,10(9),685-697)。寻找与GABAA受体相互作用且α1-GABAA受体相关副作用更少的新化合物将有巨大的治疗潜力。
发明内容
根据本申请的一个方面，提供了一种式(1)所示化合物、其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、多晶型物或溶剂合物：
其中，
R1为取代或者未取代的杂环与杂环形成的稠合基；
R2选自H、卤素、OH、C1-C6烷氧基或CN；
R3选自H、取代或者未取代的直链或者支链C1-C6烷基或取代或者未取代的取代的C3-C6环烷基。
除特别指明外，下列定义用于说明和定义在本文中用于描述本申请时使用的各种术语的意义和范围。
无论是单独出现还是组合出现，一般术语的下列定义均适用。
本申请中使用的命名规则基于IUPAC规则。
除特别说明外，术语“取代的”是指指定的基团或部分可以具有1、2、3、4、5或6个取代基。当基团上可以具有多个取代基并且给出了多种可能的取代基时，所述取代基独立选择，不必是相同的。
术语“未取代的”是指指定的基团上不具有取代基。
术语“任选取代的”是指指定的基团是未取代的或者是被一个或多个独立选自可能的取代基所取代的。
当指明取代基的数目时，术语“一个或多个”是指一个取代至取代的最多可能的数目，即取代一个氢至所有的氢均被取代基取代。除特别指明外，优选1、2、3、4或5个取代基。
具体的，R1和R3的基团定义中，“取代的”意为所述基团上一个或多个氢被选自C1-C4的烷基、C1-C4的烷氧基、C1-C4的卤素取代烷基或者卤素取代。
术语“卤素”是指氟、氯、溴和碘，优选氟。
本申请的化合物可以含有不对称中心或手性中心，并且因此存在不同的立体异构体形式。本申请化合物的所有立体异构体形式，包括但不局限于，非对映体、对映体和位阻异构体，以及它们的混合物例如外消旋混合物，将形成本申请的一部分。在本文中，当任何特定手性原子的立体化学未确定时，所有立体异构体均被考虑。此外，本申请涉及所有的几何和位置异构体。本申请化合物可以以不同的互变异构体形式存在，并且所有这些形式均包括在本申请范围内。本申请化合物的所有立体异构体预期包括混合物形式或纯的或基本上纯的形式。可以通过物理方法例如分步结晶、非对映体衍生物的分离或结晶、或者通过手性柱层析分离来拆分。
术语“前体药物”是式(1)的化合物的功能性衍生物，该衍生物在体内容易转化成式(1)的化合物。可以通过本领域技术人员众所周知的常规技术选择和制备合适该衍生物，例如参见Design of Prodrugs,ed.H.Bundgaard,Elsevier,1985。
本文所用术语“药学上可接受的盐”，是指本申请化合物的药学上可接受有机或无机盐。示例性的盐包括但是不局限于硫酸盐、枸橼酸盐、乙酸盐、草酸盐、氯化物、溴化物、碘化物、硝酸盐、酸式硫酸盐、异烟酸盐、乳酸盐、水杨酸盐、酸式枸橼酸盐、琥珀酸盐、马来酸盐、延胡索酸盐、葡萄糖酸盐、甲酸盐、甲磺酸盐和巴莫酸盐。“药学上可接受的盐”可涉及包括另一分子例如马来酸盐或其他平衡离子。平衡离子在母体化合物中稳定电荷。“药学上可接受的盐”可以有多于一个的荷电原子，多个荷电原子可具有多个平衡离子。
如果本申请化合物是碱，需要的“药学上可接受的盐”可通过适宜的方法制备，例如，用以下的无机酸处理该游离碱：盐酸、氢溴酸、硫酸、硝酸、磷酸；或者用如下的有机酸：乙酸、马来酸、琥珀酸、扁桃酸、富马酸、丙二酸、丙酮酸、水杨酸、吡喃糖苷基酸如葡萄糖醛酸或半乳糖醛酸、α-羟基酸如枸橼酸或酒石酸、氨基酸如谷氨酸、芳香族酸如苯甲酸或肉桂酸、磺酸如甲磺酸或对甲苯磺酸。
如果本申请化合物是酸，需要的“药学上可接受的盐”可通过适宜的方法制备，例如，用如下的无机碱或者有机碱处理该游离酸：胺、碱金属氢氧化物或碱土金属氢氧化物等。适宜的盐的示例性的示例包括但是不限于由氨基酸得到的有机盐，伯、仲、叔胺盐，以及环状胺例如哌啶、吗啉和哌嗪的盐，以及由钠、钙、钾、镁、锰、铁、铜、锌、铝和锂得到的无机盐。
本申请的化合物可以以从完全无定形到完全结晶的连续固态存在。术语“无定形”是指材料在分子水平上缺少长程有序性的状态，并且取决于温度，可以表现出固体或液体的物理性质。典型地，这样的材料不给出明显的X射线衍射图案，并且在显示出固体性质的同时，更正式地描述为液体。在加热时，发生从固体性质到液体性质的变化，其特征在于状态的变化，通常是二级(“玻璃化转变”)。 术语“晶体”指的是一种固相，其中材料在分子水平上具有规则有序的内部结构，并给出具有确定峰的独特的X射线衍射图。这样的材料在充分加热时也将表现出液体的性质，但是从固体到液体的变化的特征在于相变，通常是一级(“熔点”)。
本文所用术语“多晶型物”是指本申请的某些化合物在固体状态下由于存在两种或两种以上不同分子排列而产生的不同固体结晶相。本申请的某些化合物可以存在多于一种晶型，本申请旨在包括各种晶型及其混合物。
本文所用术语“溶剂合物”是指一个或多个溶剂分子与本申请化合物的结合物或络合物。形成溶剂合物的溶剂的示例包括但是不局限于水、异丙醇、乙醇、甲醇、二甲亚砜、乙酸乙酯、乙酸和乙醇胺。本申请化合物可以以非溶剂化形式存在，也可以与药学上可接受的溶剂如水、乙醇等以溶剂化形式存在，所以本申请将包括溶剂化和非溶剂化的形式。
本申请的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素，术语“同位素体”具有相同的原子数，但是其原子质量或质量数不同于在自然界中占优势地存在的原子质量或质量数。例如，可用放射性同位素标记化合物，比如氘(2H)、氚(3H)、碘-125(125I)或C-14(14C)。本申请的化合物的所有同位素组成的变换，无论放射性与否，都包括在本申请的范围之内。同位素变体可能会提高某些治疗优点，比如氘富集可以增加体内半衰期或减少剂量需求，或者可以提供可用作生物样品的表征的标准品化合物。通过本领域技术人员众所周知的常规技术，或者通过与在本文的路线和实施例中所述的那些类似的方法，使用适当的同位素富集的试剂和/或中间体，无需过多实验，可以制备式(1)内的同位素富集的化合物。
[R2]
在本申请所述化合物中，所述R2选自H、卤素、OH、C1-C6烷氧基或CN；优选的，所述R2优选为H或卤素，更优选为H或F。
[R3]
在本申请所述化合物中，所述R3选自H、取代或者未取代的直链或者支链C1-C6烷基或取代或者未取代的C3-C6环烷基。在R3中，所述C1-C6烷基可以是直链，也可以是支链，示例性的C1-C6烷基包括甲基、乙基、正丙基(1-丙基)、异丙基(2-丙基，1-甲基乙基)、正丁基(1-丁基)、仲丁基(2-丁基，1-甲基丙基)、异丁基(2-甲基丙基)或Fe/F-丁基(1，1-二甲基乙基)。上述烷基上1至多个氢原子可以被甲基、乙基、正丙基、甲氧基、乙氧基、丙氧基、氟代甲基、氟代乙基、氟、氯、溴等取代基所取代。
术语“环烷基”是指单价饱和的环状烃基，在R3中，示例性的C3-C5环烷基包括环丙基、环丁基或环戊基。
在R3中，示例性的C3-C6环烷基包括1-甲基环丙基、2-甲基环丙基、1-甲基环丁基、2-甲基环丁基、3-甲基环丁基、1-甲基环戊基、2-甲基环戊基或3-甲基环戊基。上述环烷基上1至多个氢原子可以被甲基、乙基、正丙基、甲氧基、乙氧基、丙氧基、氟代甲基、氟代乙基、氟、氯、溴等取代基所取代。
术语“烷氧基”是指烷基氧基，在R3中，示例性的C1-C6烷氧基包括甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、仲丁氧基等。上述烷氧基上1至多个氢原子可以被甲基、乙基、正丙基、甲氧基、乙氧基、丙氧基、氟代甲基、氟代乙基、氟、氯、溴等取代基所取代。
在本申请一种可选地实施方案中，R3选自直链或者支链C1-C6烷基、C1-C6烷氧基，更优选为C2-C4烷基，更优选乙基或异丙基。
[R1]
在本申请方案中，所述R1为杂环与杂环稠合基，两个杂环独立地是指具有杂原子的饱和或部分不饱和的单环或多环基团，可选地，所述杂原子为N、O或S。可选地，两个杂环独立地为含有1-3个选自N、O或S的环杂原子的3-7元饱和或部分不饱和的单环或多环基团。可选地，S和O可以以-SO或者SO2存在。
可选地，两个杂环独立地为含有1个、2个或3个选自N、O或S的环杂原子的5-7元饱和或不饱和的单环基。可选地，两个杂环独立地为含有1个、2个或3个选自N或O的环杂原子的5-6元饱和或不饱和的单环基。
可选地，R1具有式(2)、(3)、(4)、(5)、(6)或(7)所示的结构：
具体的，
所述R4选自氢、C1-C6烷基、C3-C6环烷基、C1-C6烷氧基、C1-C6烷硫基或者C1-C6烷基磺酰基；上述基团(包括涉及的烷基、环烷基、烷氧基、烷硫基或者烷基磺酰基)可选择的未被取代或者彼此独立的被1-4个分别选自卤素、羟基、C1-C3烷基、卤代C1-C3烷基、C1-C3烷氧基或者卤代C1-C3烷氧基中的至少一种所取代；
所述R5选自氢、卤素、羟基、氧代、C1-C6烷基酰基、C1-C6烷基酰胺基、C1-C6烷氧基亚胺基、C1-C6烷基、C2-C6烯基、C2-C6炔基、C3-C6环烷基、C1-C6烷氧基、C6-C10的芳基、含有1-3个杂原子的5-10元的杂芳基、含有1-3个杂原子的C1-C6的非芳族杂环；上述基团(包括涉及的所述烷基酰基、烷基酰胺基、烷氧基亚胺基、烷基、烯基、炔基、环烷基、烷氧基、芳基、杂芳基、非芳族杂环烷基)可选择的未被取代或者彼此独立的被1-4个分别选自卤素、羟基、C1-C3烷基、卤素取代的C1-C3烷基、C1-C3烷氧基或者含有1-3个N或O的杂原子的C1-C6的3-7元杂环烷基中的至少一种所取代；
所述A环是指具有含N的5-7元饱和或部分不饱和的单环基团，m为1、2或3，n为1或2，优选的，m为1及n为1，表示连接位点。
R1通过与式(1)化合物的苯基连接。
R4或者R5为杂环上的取代基，可以与杂原子连接，也可以与碳原子连接。
R5基团中，杂原子选自N、O或S。
在某些可选的实施方式中，R4选自氢、甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、乙烯基、乙炔基、环丙基、环丁基、甲氧基、乙氧基、正丙氧基、-CH2-O-CH3、甲基磺酰基或者乙基磺酰基；上述基团可选择的未被取代或者彼此独立的被C1-C3烷氧基或卤素取代。优选地，R4选自H、C1-C3烷基、C1-C3烷氧基取代的C1-C3烷基或者C1-C3烷氧基，最优选甲氧基甲基或者甲氧基。
在某些可选的实施方式中，R5选自氢、F、Cl、Br、羟基、甲酰基、乙酰基、甲酰胺基、乙酰胺基、甲氧亚胺基、乙氧亚胺基、甲基、乙基、异丙基、环丙基、环丁基、环戊基、环己基、甲氧基、乙氧基、苯基、苯甲基、四氢呋喃基、六氢吡喃基、六氢吡啶基、氧杂环丁烷、氮杂环丁烷；上述基团可选择的未被取代或者彼此独立的被1-4个分别选自F、Cl、Br、羟基、甲氧基、甲基、四氢吡咯、吗啉基或者-CH2-CF3中的至少一种所取代。
在可选的实施方式中，R5选自H、甲基、乙基、异丙基、 环丙基、氟代环丙基、 或者
可选地，A环是指含有1个、2个或3个N的环杂原子的5-6元饱和或不饱和的单环基。
在某些可选的实施方式中，当R1具有式(2)或(7)所示的结构时，A环具有如下结构：
或者
表示连接位点；所述R5如本申请前述任一项所定义。
在某些可选的实施方式中，当R1具有式(3)、(4)、(5)或(6)所示的结构时，A环具有如下结构：
或者
表示连接位点；所述R5如本申请前述任一项所定义。
在本申请方案中，可选地，所述R1为取代或者未取代的吡啶稠杂环；更优选的，所述R1为取代或者未取代的吡啶并三氮唑或者取代或者未取代的吡啶并咪唑。
作为更优选的方案，所述R1选自如下化合物中的任意一种：
或者
其中，表示连接位点。
所述R4和R5如本申请前述任一项所定义。
作为可选的结构，所述R1选自如下结构中的任意一种：
或者并且，
所述R4选自甲氧基C1-C3烷基或C1-C3烷氧基；更优选为甲氧基甲基或者甲氧基；
所述R5选自H、甲基、乙基、异丙基、环丙基、氟代环丙基、
更优选的，本申请所述化合物具有如下表1所示结构。
表1优选化合物结构






本申请还提供了一种药物混合物，所述药物混合物包含选自下组的两种或两种以上的化合物：本申请任一项所述的化合物或其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、同位素体、多晶型物或溶剂合物。
本申请还提供了一种药物组合物，其包含本申请任一项所述的化合物或其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、同位素体、多晶型物或溶剂合物中的至少一种，并可选择的添加药学上可接受的载体和/或辅助剂。
本申请任一项所述的化合物或其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、多晶型物、溶剂合物、所述的药物混合物，或所述的药物组合物在制备治疗或预防与GABAA受体有关的疾病的药物中的用途。
进一步地，所述与GABAA受体有关的疾病选自如下至少一种：疼痛、阿尔茨海默氏病、多梗塞性痴呆和中风。
进一步地，所述的疼痛是神经病理性疼痛、炎性疼痛和癌性痛。
进一步地，所述的疼痛选自：头痛，面部痛、颈痛、肩痛、背痛、胸痛、腹痛、背部痛、腰痛、下肢痛、肌肉与骨骼疼痛、血管疼痛、痛风、关节炎疼痛、内脏疼痛、感染性疾病导致的疼痛、多骨疼痛、镰刀细胞贫血、自身免疫性疾病、多发性硬化或炎症有关的疼痛，损伤或手术引起的慢性疼痛、伤害感受性疼痛、疼痛性糖尿病、三叉神经痛、腰部或子宫颈神经根病痛、舌咽神经痛、自主神经反射性疼痛、反射性交感神经营养不良、神经根撕脱、癌症、化学损伤、毒素、营养缺乏、病毒或细菌感染或退行性骨关节病有关的疼痛。
本申请还提供了一种治疗或预防与GABAA受体有关的疾病的方法，向患者施用有效剂量的本申请任一项所述的化合物或其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、多晶型物、溶剂合物、所述的药物混合物，或所述的药物组合物。
本申请还提供了一种治疗或预防疼痛、阿尔茨海默氏病、多梗塞性痴呆或中风的方法，向患者施用有效剂量的本申请任一项所述的化合物或其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、多晶型物、溶剂合物、所述的药物混合物，或所述的药物组合物。
本申请还涉及如上所述的式(1)化合物的制备方法，包括：

注：R3为乙基或者异丙基实验操作相同。
Z-1a：5-氯-N3-乙基哒嗪-3,4-二胺
将3,5-二氯哒嗪-4-胺(80.5g,0.5mol)和乙胺醇溶液(30％,600ml)放在高压釜中，150摄氏度反应16小时。反应液降至室温后有固体析出，过滤，滤饼用二氯甲烷洗(300ml)，母液浓缩(不蒸干)，有固体析出，再过滤，反复3次，收集所有的滤饼得到粗产物(含乙胺盐酸盐)。粗产物中加水搅拌打浆，过滤，滤饼旋干得到65g(76.5％yield)淡黄色固体目标产物。LC-MS:m/z[M+H]+＝173.
Z-2a：4-氯-7-乙基-7H-咪唑并[4,5-c]哒嗪
将5-氯-N3-乙基哒嗪-3,4-二胺(30g,0.17mol)加入原甲酸三甲酯(600ml)中。120摄氏度反应4小时。反应液直接旋干得到粗产物，粗产物加二氯甲烷甲醇溶解拌样过柱子(二氯甲烷,二氯甲烷/甲醇＝50/1,V/V).得到20g(63％yield)黄色固体目标产物LC-MS:m/z[M+H]+＝183.
Z-3a：4-(3-氯-4-氟苯基)-7-乙基-7H-咪唑并[4,5-c]哒嗪
将4-氯-7-乙基-7H-咪唑并[4,5-c]哒嗪(20g,108mmol)，(3-氯-4-氟苯基)硼酸(19.08g,108mmol)，碳酸钠(24g,216mmol),[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(9.2g,10.8mmol)加入1,4-二氧六环/水(160ml/40ml)中，90摄氏度反应2小时。反应液直接柱层层析纯化(石油醚/乙酸乙酯＝10/1，5/1,1/1,乙酸乙酯,V/V.)得到黄色固体标题化合物(26g，85.7％yield).LC-MS:m/z[M+H]+＝277.
Z-4a-1：7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼硼烷-2-基)苯基)-7H-咪唑并[4,5-c] 哒嗪
将4-(3-氯-4-氟苯基)-7-乙基-7H-咪唑并[4,5-c]哒嗪(15g,54mmol),联硼酸频那醇酯(27.6g,108mmol),乙酸钠(13g,163mmol),2-二环己基磷-2,4,6-三异丙基联苯(2.7g,5.4mmol),三(二亚芐基丙酮)二钯(5g,5.4mmol)加入无水1,4-二氧六环(150ml)中，110摄氏度搅拌过夜。反应液直接柱层层析纯化(二氯甲烷/无水甲醇＝50/1)得到淡黄色固体标题化合物(12g，60％yield)。1H NMR(400MHz,CHLOROFORM-d)ppm 1.40(s,12H)1.69(t,J＝7.09Hz,3H)4.58(q,J＝7.34Hz,2H)7.26(d,J＝6.36Hz,1H)8.29(s,1H)8.45(d,J＝1.96Hz,2H)9.39(s,1H).LC-MS:m/z[M+H]+＝287,369.
Z-4a-2：(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸
将7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(12g，32.5mmol)溶于200mL盐酸水溶液(2M)中，室温下搅拌2小时，冻干得标题化合物(9.3g，粗品)，外观为白色固体。
本申请的式(1)化合物及其药学上可接受的盐可以通过上述方法制备。
如果没有在实施例中描述其制备方法，那么式(1)化合物及其中间体产物可以根据类似的方法或者根据前述方法制备。本领域已知的原料可以得自商业，或者可以根据本领域已知的方法或已知方法的类似方法制备。
可以理解，本申请的式(1)化合物可以在官能团上衍生，从而得到能够在体内再转化为母体化合物的衍生物。
因此，本申请还涉及药用组合物，该药用组合物包括如上文所定义的化合物或其药学上可以接受的盐或其前药和药学上可接受的载体和/或辅助剂。
同样，本申请还包括如上所述的化合物或者组合物，用作制备治疗或预防与α2/3-GABAA受体 有关的疾病的药物的用途，尤其是治疗或预防下列疾病：疼痛、癫痫、焦虑、瘙痒和抑郁。
优选治疗或预防疼痛。
特别优选治疗或预防神经病理性痛、炎性疼痛和癌性痛。
如本文所用，“癌性痛”是指恶性肿瘤在其发展过程中出现的疼痛，癌性痛发生目前认为有三种机制，即：癌症发展直接造成的疼痛、癌症治疗后造成的疼痛和癌症患者并发疼痛性疾病。
如本文所用，“神经病理性痛”是为由神经系统原发性损害和功能障碍所激发或引起的疼痛。
如本文所用，“炎性疼痛”是局部急性炎症或是慢性炎症刺激神经所致的疼痛。
如本文所用，“治疗”也包括预防性给药，一旦所述病症建立后缓解或消除所述病症。
如本文所用，“患者”定义为任何温血动物，例如不限于小鼠、豚鼠、狗、马或人，所述患者最好是人。
如本文所用，“急性疼痛”定义为由皮肤、身体结构或内脏的损伤和/或疾病发生的有害刺激而引起的疼痛，或由不产生实际组织损害的肌肉或内脏的异常功能而引起的疼痛。
如本文所用，“慢性疼痛”定义为持续超出急性疾病通常的病程或损伤治愈的合理时间，或与引起持续疼痛的慢性病理过程有关，或疼痛以一定间隔复发数月或数年，如果在应该已经达到治愈后或超过通常的治疗过程后仍存在疼痛，则认为是慢性疼痛。在疼痛需要经过的时间长度取决于疼痛的性质和与疼痛有关的治疗过程，如果疼痛超过通常的治疗过程，则疼痛是慢性的。慢性疼痛包括但不限于头痛、面部痛、颈痛、肩痛、胸痛、腹痛、背部痛、腰痛、下肢痛、肌与骨骼疼痛、与躯体病样精神障碍有关的疼痛、内脏痛、疼痛性糖尿病性神经病、血管疼痛、痛风、关节炎性疼痛、癌症疼痛、自主神经反射性疼痛、感染性疾病(如艾滋病和带状疱疹)导致的疼痛、自身免疫性疾病(风湿)导致的疼痛、急慢性炎症导致的疼痛、手术后疼痛和烧伤后疼痛。
本申请揭示的药物能有效地治疗如上定义的慢性疼痛，而且本申请揭示的药物可用于治疗伴随其他病症的痛敏，包括痛觉过敏、异常性疼痛、痛觉增强和疼痛记忆增强，该申请将改善对其疼痛的治疗。
如本文所用，“头痛”可分为原发性头痛和继发性头痛，原发性头痛包括紧张性头痛、偏头痛和丛集性头痛，而继发性头痛是由于其他疾病引起的。头面部的痛敏感组织发生病变或受到刺激时，可引起各种头痛，这些痛敏感组织包括分布于头皮、面部、口腔及咽喉等，由于它们主要是头部的肌肉或血管，含有丰富的神经纤维，对疼痛比较敏感，所以当这些组织受到伤害时可引起头痛。
如本文所用，“面部痛”包括但不限于三叉神经痛、非典型面痛、面神经麻痹和面肌痉挛。
如本文所用，“三叉神经痛”是一种独特的慢性疼痛性疾病，又称痛性抽搐，是指在三叉神经分布区域出现短暂的、阵发性的和反复发作的电击样剧烈性疼痛，或伴有同侧面肌痉挛。三叉神经痛分为原发性和继发性两种类型，原发性三叉神经痛是指临床上未发现神经系统体征，检查未发现器质性病变；继发性三叉神经痛是指在临床上有神经系统体征，检查发现有器质性病变，如肿瘤和炎症等。
如本文所用，“非典型面痛”是指由多种病因引起的疼痛。表现为持续性烧灼样疼痛、无间歇性、与特殊的动作或触发刺激无关，疼痛多为双侧、疼痛常常超出三叉神经的分布范围甚至累及颈部皮肤。病因可由鼻窦炎、恶性肿瘤、颌及颅底感染等原因刺激或损伤三叉神经而引起疼痛。
如本文所用，“颈痛、背痛、肩痛”是指由于急慢性肌肉劳损及骨关节的退行性变和外伤等导致的疼痛。引起颈、肩及上肢疼痛的常见疾病有颈肩肌筋膜炎、项韧带炎、颈椎病、肩周炎、胸廓出口综合症、肱骨外上髁炎等，或由自身免疫性疾病引起的疼痛常见于类风湿性关节炎、强直性脊柱炎和风湿性关节炎等疾病，其他可能引起颈痛、背痛、肩痛的疾病还有颈、肩部的肿瘤、神经炎、动静脉疾病和各种感染以及胸、腹腔脏器病变引起的牵涉痛等。
如本文所用，“胸、腹及背部痛”指由于胸腹部内脏、胸腹壁组织的疾病导致的疼痛，包括但不限于肋间神经痛、肋间软骨炎、心绞痛、腹痛(急性腹部内脏痛)和腰背部肌筋膜综合征。
如本文所用，“腰、下肢痛”是指下腰、腰骶、骶髂、髋、臀及下肢痛。腰和下肢痛往往并不是独立的疾病，而是多种疾病的共有特征，临床表现多样，病因十分复杂，以退行性和损伤为多，包括但不限于腰椎间盘突出、急性腰扭伤、坐骨神经痛、骨质疏松症、第三腰椎横突综合症、梨状肌综合症、膝关节骨性关节炎、尾痛症和足跟痛等涉及的疼痛。
如本文所用，“肌与骨骼疼痛”包括但不限于肌筋膜疼痛、创伤引发的疼痛和慢性区域性疼痛综 合症。
如本文所用，“疼痛性糖尿病”是指糖尿病并发的神经损伤而引起的疼痛，糖尿病中的神经损伤至少部分是由于血流减少和高血糖引起的。某些糖尿病患者不发生神经病变，而其他患者早期就发生该疾病，糖尿病性神经病痛可以分为涉及一个或者多个病灶部位的单神经病和全身性多神经病，所述多神经病可以是扩散和对称的，通常主要涉及感觉方式(Merrit’s Textbook of Neurology，第9版，LPRowland LP编辑)。糖尿病性神经病的表现可以包括植物神经功能障碍，导致包括心脏、平滑肌和腺体在内的调节障碍，引起低血压、腹泻、便秘和性无能。糖尿病神经病往往分阶段发展，早期在神经末梢区，植物神经病或感觉神经病的时发生在足部，脑神经病时发生在面部和眼周围，出现间歇性疼痛和麻刺感，在随后的阶段中，疼痛更强和经常发生，最后，当某一区域痛觉丧失时，发生为无痛性神经病，由于没有疼痛作为损伤的指示，大大增加了发生严重组织损伤的风险。
如本文所用，“内脏疼痛”包括但不限于刺激性肠综合征(IBS)，伴有或不伴有慢性疲劳综合征(CFS)、炎性肠病(IBD)和间质性膀胱炎的疼痛。
如本文所用，“血管疼痛”是由以下一种或多种因素产生的疼痛。第一，组织的灌注不当。引起暂时或连续的局部缺血，诸如在运动期间发生肢体肌肉中的局部缺血；第二，迟发性变化。例如在皮肤或腹部内脏中的溃疡或坏疽；第三，大血管口径的突然或加速变化。例如动脉瘤发生的变化；第四，主动脉破裂。结果是血液溢出，刺激腹膜或胸膜壁层中的伤害感受纤维；第五，由于动脉内注射严重刺激动脉内皮而引起的强痉挛；第六，静脉血回流的损害，结果是迅速扩张筋膜隔室的大量水肿(Bonica等，The Management of Pain，第一卷(第二版)，Philadelphia；Lea&Feboger，1990)。实例包括但不限于闭塞性动脉硬化、闭塞性血栓脉管炎、急性动脉闭合、栓塞、先天性动静脉瘤、血管痉挛性疾病、Rayaud病、手足发绀、急性静脉闭合、血栓静脉炎、静脉曲张和淋巴水肿。
如本文所用，“自主神经反射性疼痛”是指由“反射性交感神经萎缩征”导致的疼痛。反射性交感神经萎缩征是指机体遭受急慢性损伤后，有剧烈的自发疼痛，对触觉和痛觉过敏，可伴有浮肿和血行障碍，随后可出现皮肤及肌肉骨骼的营养障碍和萎缩等症状。
如本文所用，“术后疼痛”是指机体对疾病本身和手术造成的组织损伤的一种复杂的生理反应，它表现为心理和行为上的一种不愉快的经历。
如本文所用，“关节炎性疼痛”包括但不限于骨关节炎、类风湿性关节炎、关节强直性脊椎炎、牛皮癣性关节病、痛风、假痛风、传染性关节炎、腱炎、粘液囊炎、骨损害和关节软组织炎症等疾病导致的疼痛。
如本文所用，“带状疱疹后的神经痛”是指带状疱疹的皮疹愈合后，在原来皮疹区的皮下长期存在的剧烈疼痛。
如本文所用，“伤害感受性疼痛”是由刺激伤害性感受器传入的组织损害过程引起的疼痛，或是由伤害性感受器延长的兴奋引起的疼痛。由伤害感受器延长的兴奋引起的疼痛可以是由于伤害感受器的持久有害刺激或其敏化或这两者共同引起，或它们可以由这些因素引起，并由其持久性、各种反射机制和其他因素而延长。
本申请提供含治疗有效量的α2/3-GABAA正向别构调节剂的药物化合物的用途。尽管用于本申请治疗的α2/3-GABAA正向别构调节剂可以原料化合物的形式给药，但优选将活性成分，任选地以生理上可接受的盐的形式，与一种或多种添加剂、赋形剂、载体、缓冲剂、稀释剂和/或其它常规的药物辅料一起混合成药物组合物。
在可选的实施方案中，本申请提供含α2/3-GABAA正向别构调节剂的药物组合物，其中α2/3-GABAA正向别构调节剂与一种或多种药学上可接受的载体、和任选地与其它本领域已知的或使用的治疗性的和/或预防性的组份混合。该载体必须是“可接受的”，即与制剂中的其它成分相容且不会对其接受者有害。
因此可将用于本申请的化合物与常规的添加剂、或稀释剂一起制成药物组合物及其单位剂量的形式。诸如此类的形式包括固体(尤其是片剂、填充胶囊、粉末以及丸剂的形式)、和液体(尤其是水溶液或非水溶液、混悬液、乳剂、酏剂)、和填充上述形式的胶囊、所有口服给药的形式、直肠给药的栓剂、以及肠胃外给药的无菌可注射的溶液。诸如此类药物组合物及其单位剂量形式可包括常规比例的常规成分、含或不含另外的活性化合物或成分，这类单位剂量形式可含与所需的每日应用剂量范围相当的任何合适的有效量的活性成分。
用于本申请的化合物可以各种的口服的和胃肠外的剂型给药。对本说领域的技术人员来说下述的剂型可含作为活性成分的本申请的化合物或其药学上可接受的盐。
为将用于本申请的化合物制成药物组合物，药学上可接受的载体可以是固体或者液体。固体形式的制剂包括粉末、片剂、九剂、胶囊、扁囊剂、栓剂、以及可分散的颗粒剂。固体载体可以是一种或多种还起稀释剂、调味剂、增溶剂、润滑剂、悬浮剂、粘合剂、防腐剂、片剂崩解剂、或囊化材料作用的物质。
粉末中，载体为细分的固体，它与细分的活性成分混合。
片剂中，活性成分与具有必要的粘合性能的载体以适当的比例混合并压缩成所需的形状和大小。
粉末和片剂优选地含5％或10％到约70％的活性化合物。合适的载体为碳酸镁、硬脂酸镁、滑石粉、糖、乳糖、果胶、糊精、淀粉、明胶、西黄蓍胶、甲基纤维素、羧甲基纤维素钠、低熔点的蜡、可可脂等等。术语“制剂”包括含与作为载体的囊化材料配制的活性化合物，囊化材料提供囊，其中含或不含载体的活性成分被载体包围，这样与其结合在一起。同样地，制剂包括扁囊剂和锭剂(lozenges)。片剂、散剂、胶囊、丸剂、扁囊剂和锭剂可以用作适合于口服给药的固体形式。
为制备栓剂，首先将低熔点的蜡，如脂肪酸甘油酯或可可脂的泥合物熔化，然后通过搅拌将活性成分均匀地分散于其中。然后将该熔化的均匀混合物倒入适当大小模具中，让其冷却并由此固化。
适合于阴道给药的组合物可以阴道栓剂、棉塞、乳膏剂、凝胶剂、糊剂、泡沫或喷雾剂的形式存在，组合物除含活性成分外还含本领域已知的合适的载体。
液体制剂包括溶液、混悬液和乳剂，例如，水溶液或水-丙二醇溶液。例如，肠胃外注射液体制剂可以配制成水-聚乙二醇的溶液。
因此用于本申请的化合物可配制成用于肠胃外给药(例如注射，如快速浓注或连续输注)的制剂，和可以与添加的防腐剂一起以单位剂量的形式存在于安瓿、预填充的注射器、小体积的输液袋中或多剂量容器中。该组合物可采取油性或水性载体的混悬液、溶液或乳剂的形式，并可含制剂成分，如悬浮剂、稳定剂和/或分散剂。另外，活性成分可以是粉末的形式，可由灭菌的固体无菌分离或由溶液冻干获得，用于临用前与合适的载体如无菌的、无热原的水重建。
适合于口服给药的水溶液可通过将活性成分溶解于水中和加入所需的着色剂、调味剂、稳定剂和增稠剂来制备。
适合于口服给药的水悬浮液可通过将细分的活性成分分散于含粘性物质，如天然的或合成的树胶、树脂、甲基纤维素、羧甲基纤维素钠、或其它公知的悬浮剂的水中而制备。
还包括为在临用前不久转化为用于口服给药的液态制剂而设计的固体制剂。这类液体制剂包括溶液、混悬液和乳剂。除活性成分之外，这类制剂可含着色剂、调味剂、稳定剂、缓冲剂、人造的和天然的甜味剂、分散剂增稠剂、增溶剂等。
为了局部施用到表皮，可将本申请的化合物配制成软膏剂、乳膏剂或洗剂或透皮贴剂。例如，软膏剂和乳膏剂可用水性或油性基质外加合适的增稠剂和/或胶凝剂配制而成。洗剂可用水性或油性基质配制而成，且通常还含一种或多种乳化剂、稳定剂、分散剂、悬浮剂、增稠剂或着色剂。
适合于口腔局部给药的组合物包括在调味基质通常为蔗糖和金合欢胶或西黄蓍胶中含有活性成分的锭剂(lozenges)；在惰性的基质如明胶和甘油或蔗糖和金合欢胶中含活性成分的锭剂(pastiIles)；以及在合适的液体载体中含活性成分的漱口药。
可将溶液或混悬液用常规方法例如用滴管、吸管或喷雾器直接应用到鼻腔。该组合物可以是单剂量或多剂量的形式。
呼吸道给药也可以通过气雾剂实现，其中活性成分与合适的推进剂一起装在加压的包装中，合适的推进剂包括氟氯化碳(CFC)例如二氯二氟甲烷、三氯氟甲烷或二氯四氟乙烷，二氧化碳或其它合适的气体。气雾剂还可适当地含表面活性剂，如卵磷脂。药物的剂量可通过量阀控制。
另外活性成分可以是干粉的形式，例如化合物与合适粉末基质如乳糖、淀粉、淀粉衍生物如羟丙基甲基纤维素和聚乙烯吡咯烷酮(PVP)的粉末混合物。粉末载体可方便地在鼻腔内形成凝胶。粉末组合物可以单位剂量的形式存在，例如存在于胶囊或药筒(如明胶的胶裘或药筒)中，或存在于粉末可经吸入器给药的泡罩包装中。
在用于呼吸道给药的组合物(包括鼻内用的组合物)中，通常化合物具有小的粒度，例如为5微米或更小数量级的粒度。这样的粒度可以用本领域已知的方法，例如通过微粉化获得。
需要时，可以应用适于活性成分缓释的组合物。
药物制剂可选为单位剂量形式。这类形式中，制剂被细分成合适量活性成分的单位剂量。单位剂量形式可以是封装的制剂，其中密封包装中含分离的大量制剂，如封装的片剂、胶囊和装入小瓶或安瓿中的粉末。此外，单位剂量形式可以是胶囊、片剂、扁裘剂或锭剂(lozenge)本身，或可以是任何封装形式的适量上述胶囊、片剂等。
用于口服给药的片剂或胶囊和用于静脉给药的液体以及连续的输液为优选的组合物。
关于制剂和给药技术的更详细的资料可以在Remington's Pharmaceutical Sciences(雷明顿药物科学)(Maack Publishing Co.,Easton,PA)的最新的版本上见到。
单位剂量制剂中活性组份的量可根据具体的应用和活性组份的效力而变化，可调节自0.01mg至约0.1g。例如，在医药用途中，该药物可以0.01至约100mg的胶囊每天给药三次，必要时该组合物还可以含其他相容的治疗剂。
治疗方法
在治疗用途中，用于本申请的化合物以起始剂量每天0.001mg/kg至10mg/kg体重。但是，这些剂量可以根据患者的需要、被治疗病症的严重性以及使用的化合物而变化，一般来说，开始以小于该化合物最佳剂量的较小剂量治疗，此后，小量增加此剂量达到最佳效果，方便起见，如果需要可将总日剂量再细分为一天内分次给药。
本申请的药物组合物还可同时与其它治疗疼痛、癫痫、焦虑和抑郁的药物联合使用，包括但不限于吗啡、加巴喷丁等。因此，本申请提供了一种用于治疗疼痛、癫痫、焦虑和抑郁的药物的药物，该药物不仅有效，而且没有明显的副作用，本申请的另一个目的是提供一种对于特殊病人群体，如老人、患有肝或肾功能衰退、或心脏血管性疾病的病人，具有高度安全性的药物。
本申请还提供了一种治疗或预防疾病的方法，向患者施用有效剂量的如上所述的化合物或组合物。
本申请还提供了一种治疗或预防与GABAA受体有关的疾病的方法，向患者施用有效剂量的如上所述的化合物或如上所述的组合物。
本申请还提供了一种如上所述的化合物或组合物在制备治疗或预防下列疾病的药物中的用途：疼痛、阿尔茨海默氏病、多梗塞性痴呆和中风。
所述的疼痛是神经病理性疼痛、炎性疼痛和癌性痛。可选地，所述疼痛选自：头痛，面部痛、颈痛、肩痛、背痛、胸痛、腹痛、背部痛、腰痛、下肢痛、肌肉与骨骼疼痛、血管疼痛、痛风、关节炎疼痛、内脏疼痛、感染性疾病导致的疼痛、多骨疼痛、镰刀细胞贫血、自身免疫性疾病、多发性硬化或炎症有关的疼痛，损伤或手术引起的慢性疼痛、伤害感受性疼痛、疼痛性糖尿病、三叉神经痛、腰部或子宫颈神经根病痛、舌咽神经痛、自主神经反射性疼痛、反射性交感神经营养不良、神经根撕脱、癌症、化学损伤、毒素、营养缺乏、病毒或细菌感染或退行性骨关节病有关的疼痛。
本申请还提供了一种治疗或预防疼痛、阿尔茨海默氏病、多梗塞性痴呆或中风的方法，向患者施用有效剂量的如商所述的化合物或如上所述的组合物。
有益效果：
本申请的化合物及其药学上可接受的盐或其前药具有重要的药理学性质，为α2/3-GABAA受体正向别构调节剂。本申请的化合物对α2/3-GABAA受体具有优异的亲和活性、正向调控活性，且具有良好的遗传毒安全性和成药性。因此，本申请化合物及其药学上可接受的盐或其前药可以单独使用或与其他药物组合使用，用于治疗或预防与α2/3-GABA A有关的疾病。
具体实施方式
下面对本申请的技术方案进行清楚、完整地描述，显然，所描述的实施例是本申请一部分实施例，而不是全部的实施例。基于本申请中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本申请保护的范围。其中，下述实施例中纯化步骤(例如制备薄层色谱、柱层析等)采用的溶剂比例为体积比。
实施例1
1-1：(4-溴-2-氟吡啶-3-基)甲醇
4-溴-2-氟烟醛(565mg,2.77mmol)加到四氢呋喃(3ml)中，硼氢化钠(63mg，1.67mmol)加到反应液，室温搅拌20分钟。反应液倒入甲醇(10ml)淬灭，倒入水(30ml)中，二氯甲烷萃取(30ml×3次)，有机相合并，无水硫酸钠干燥，浓缩，得粗品无色油状液体(508mg,89％)。LC-MS:m/z[M+H]+＝206,208。
1-2：(4-溴-2-肼吡啶-3-基)甲醇
(4-溴-2-氟吡啶-3-基)甲醇(505mg,2.45mmol)，水合肼(246mg,4.9mmol)加到乙醇(4ml)中，95℃反应2小时。反应液直接拌样，柱层析(二氯甲烷/甲醇＝10/1)得标题化合物白色固体(388mg,51％).LC-MS:m/z[M+H]+＝218,220。
1-3：(7-溴-3-环丙基-[1,2,4]三唑并[4,3-a]吡啶-8-基)甲醇
(4-溴-2-肼吡啶-3-基)甲醇(150mg,0.48mmol)，环丙基甲醛(50mg,0.72mmol)加到乙醇(2ml)中，90℃反应1小时，碳酸钾(66mg,0.66mmol)，碘(12mg,0.05mmol)加到反应液，室温搅拌4小时。反应液制备薄层色谱(二氯甲烷/甲醇＝25/1)得标题化合物棕色固体(65mg,51％).LC-MS:m/z[M+H]+＝268,270。
1-4：7-溴-3-环丙基-8-(甲氧基甲基)-[1,2,4]三唑并[4,3-a]吡啶
(7-溴-3-环丙基-[1,2,4]三唑并[4,3-a]吡啶-8-基)甲醇(60mg,0.22mmol)加到四氢呋喃(2ml)中，氢化钠(8mg,0.26mmol)加到反应液，室温搅拌10分钟，碘甲烷(47mg,0.33mmol)加到反应液中，室温搅拌30分钟。反应液制备薄层色谱(二氯甲烷/甲醇＝30/1)得标题化合物棕色固体(55mg,89％).LC-MS:m/z[M+H]+＝282,284。
化合物1：4-(3-(3-环丙基-8-(甲氧基甲基)-[1,2,4]三唑并[4,3-a]吡啶-7-基)-4-氟苯基)-7-乙基-7H- 咪唑[4,5-c]哒嗪
7-溴-3-环丙基-8-(甲氧基甲基)-[1,2,4]三唑并[4,3-a]吡啶(30mg,0.11mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(31mg,0.11mmol)，碳酸钠(23mg,0.22mmol)，二(三苯基膦)二氯化钯(8mg,0.01mmol)加到四氢呋喃(2ml)和水(0.1ml)中，氩气保护下，80℃反应16小时。反应液制备薄层色谱(二氯甲烷/甲醇＝15/1)得标题化合物棕色固体(8mg,16％).
1H NMR(400MHz,CHLOROFORM-d)9.40(s,1H),8.42(d,J＝6.8Hz,2H),8.28(s,1H),8.16-8.10(m,1H),7.46-7.38(m,1H),6.93(br.s.,1H),4.83(br.s.,2H),4.59(d,J＝7.3Hz,2H),3.41(s,3H),2.13-2.00(m,1H),1.71-1.68(m,3H),0.88(br.s.,4H).LC-MS:m/z[M+H]+＝444.
实施例2
2-1：5-溴-4-甲氧基吡啶-2-胺
将2-氨基-4-甲氧基吡啶(5g,40mmol)，溶于无水乙腈(80.0mL)中，并分批加入N-溴代丁二酰亚胺(NBS,7g,40mmol)，0℃搅拌2小时，用饱和碳酸氢钠水溶液(100mL)淬灭后，分层，收集有机层，水相再用二氯甲烷(100mL×3)萃取，旋干后得到粗品，正相硅胶柱层析(石油醚：乙酸乙酯＝10:1～2:1)纯化后，得到标题产物(4g，58％)白色固体。LC-MS:m/z[M+1]+＝203
2-2：6-溴-2-(1-氟环丙基)-7-甲氧基咪唑并[1,2-a]吡啶
将5-溴-4-甲氧基吡啶-2-胺(500mg,2.5mmol)，2-氯-1-(1-氟环丙基)乙-1-酮(670mg,5mmol)，碳酸铯(1.6g,5mmol)溶于乙醇(10.0mL)的溶液中，在微波反应器中加热到100℃反应1小时，将反应液经过制备HPLC纯化得到标题产物(100mg,27％)白色固体。LC-MS:m/z[M+1]+＝285
化合物2：7-异丙基-4-(4-氟-3-(2-(1-氟环丙基)-7-甲氧基咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑 并[4,5-c]哒嗪
将6-溴-2-(1-氟环丙基)-7-甲氧基咪唑并[1,2-a]吡啶(100.0mg,0.36mmol)，(5-(7-异丙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(105mg,0.36mmol)，碳酸铯(210mg,0.54mmol)，[1,1-双(二叔丁基膦)二茂铁]二氯化钯(II)(16.5mg,0.036mmol)，悬浮于1，4-二氧六环/水(5.5mL,v/v＝10/1)的溶液中，加热到100℃反应2小时。反应液用乙酸乙酯(50mL)稀释，用饱和氯化钠水溶液(50mL×3)水洗，有机相干燥，浓缩，再用制备HPLC纯化得到标题产物(30mg,17％)白色固体。
1H NMR(400MHz,CDCl3)9.64(s,1H),9.06(s,1H),8.98(s,1H),8.70–8.62(m,1H),8.58(dd,J＝7.0,2.2Hz,1H),8.25(s,1H),7.65(t,J＝9.2Hz,1H),7.30(s,1H),5.22–5.02(m,1H),4.02(s,3H),1.67-1.63(m,8H),1.42(q,J＝8.5Hz,2H).LC-MS:m/z[M+1]+＝461
实施例3和实施例4

3-1：2-氟-4-碘-3-甲氧基吡啶
将2-氟-3-甲氧基吡啶(5g,39.3mmol)溶于无水四氢呋喃(150mL)中，并于氮气保护下将反应液冷却至-60℃,缓慢滴加正丁基锂(2.5M)(23.6mL，59mmol)，控制温度在-60～-65℃，用时10min，并于-60℃搅拌1h。缓慢加入碘(11g,43mmol)的无水四氢呋喃溶液(100mL)，保持温度在-60℃左右，用时2h，然后于-60℃反应2h，加入饱和氯化铵水溶液(200mL)淬灭反应，用乙酸乙酯(100mL×3)萃取，饱和食盐水(100mL)洗涤，无水硫酸钠干燥，浓缩后正相纯化(石油醚：乙酸乙酯＝3：1)得到标题产物(6.8g，白色固体)产率68％。1H NMR(400MHz,CDCl3)7.56(m,2H),3.99(s,3H).
3-2：2-肼基-4-碘-3-甲氧基吡啶
将2-氟-4-碘-3-甲氧基吡啶(6.8g,26.88mmol)溶于乙醇(68.0mL)中，并加入水和肼(2.7g,53.8mmol)，于90℃反应16小时。浓缩后加入水(100mL)，搅拌30min，过滤，收集滤饼真空干燥后得到标题产物(5.5g，白色固体)产率78％。LCMS:m/z[M+1]+＝266
3-3：N'-(4-碘-3-甲氧基吡啶-2-基)-3-甲氧基环丁烷-1-碳酰肼
将2-肼基-4-碘-3-甲氧基吡啶(300mg,1.13mmol)，3-甲氧基环丁烷-1-羧酸(147mg,1.13mmol)，2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(472.6mg,1.24mmol)溶于无水二氯甲烷(12mL)中，并于氮气保护下缓慢滴加二异丙基乙基胺(366mg，2.83mmol)，并于30℃搅拌2h。将反应液倒入冰水中，用二氯甲烷(100ml×3)萃取，浓缩后反相纯化得到标题产物(270mg，黄色油状物)产率71％。LCMS:m/z[M+1]+＝378
3-4：7-碘-8-甲氧基-3-(3-甲氧基环丁基)-[1,2,4]三唑并[4,3-a]吡啶
将N'-(4-碘-3-甲氧基吡啶-2-基)-3-甲氧基环丁烷-1-碳酰肼(270mg，0.72mmol)溶入到无水四氢呋喃(10ml)中，并加入Burgess试剂(N-(三乙基铵磺酰)氨基甲酸甲酯，512mg,2.15mmol)，并将此反应于氮气保护下于60℃反应3h。将反应液倒入水(50ml)中。用乙酸乙酯(50ml×3)萃取，浓缩后得到一粗品，HPLC制备纯化后得到标题产物(170mg，白色固体)产率66％。LCMS:m/z[M+1]+＝360
化合物3：rel-4-(4-氟-3-(8-甲氧基-3-((1s,3s)-3-甲氧基环丁基)-[1,2,4]三唑并[4,3-a]吡 啶-7-基)苯基)-7-异丙基-7H-咪唑并[4,5-c]哒嗪
化合物4：rel-4-(4-氟-3-(8-甲氧基-3-((1r,3r)-3-甲氧基环丁基)-[1,2,4]三唑并[4,3-a]吡 啶-7-基)苯基)-7-异丙基-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-碘-8-甲氧基-3-(3-甲氧基环丁基)-[1,2,4]三唑并[4,3-a]吡啶(100.0mg,0.278mmol)，(2-氟-5-(7-异丙基-7H-咪唑并[4,5-c]哒嗪-4-基)苯基)硼酸(100.3mg,0.334mmol)为原料，得到两标题的混合物，外观为白色固体。再进行手性拆分得实施例3和实施例4。拆分方法：仪器：MG Ⅱ preparative SFC(超临界流体制备色谱仪，SFC-1)；色谱柱：ChiralPak AD,250×30mm I.D.,10μm；流动相：A为CO2，B为异丙醇；B为55vt％；流速：80mL/min；背压：100bar；柱温：38℃；波长：220nm。
实施例3：第一个峰，14mg，白色固体，产率10％。
1H NMR(400MHz,DMSO-d6)9.58(s,1H),8.95(s,1H),8.64–8.44(m,2H),8.13(d,1H), 7.67–7.46(m,1H),7.03(d,1H),5.24–4.92(m,1H),4.33(d,J＝3.2Hz,3H),4.22–3.88(m,2H),3.21(s,3H),2.72(m,2H),2.59–2.49(m,2H),1.68(d,J＝6.8Hz,6H).LCMS:m/z[M+H]+＝488
实施例4：第二个峰，38mg,白色固体，产率28％。
1H NMR(400MHz,DMSO-d6)9.58(s,1H),8.95(s,1H),8.65–8.43(m,2H),8.19(d,1H),7.70–7.44(m,1H),7.04(d,1H),5.22–5.00(m,1H),4.33(s,3H),4.02(t,J＝6.9Hz,1H),3.60(d,J＝7.9Hz,1H),3.21(s,3H),2.97–2.75(m,2H),2.32(dd,J＝10.5,8.8Hz,2H),1.68(d,J＝6.8Hz,6H).LCMS:m/z[M+H]+＝488.
实施例5
5-1：4-(甲氧基亚甲基)-2-甲基四氢-2H-吡喃
将(甲氧基甲基)三苯基氯化膦(22g,64.4mmol)悬浮于无水四氢呋喃(180mL)中,并于氮气保护下将反应液冷却至-10～-20℃，缓慢滴加双三甲基硅基胺基锂(1M，64.3mL，64.4mmol)，控制温度在-10～-20℃，用时30min，并于-10～-20℃搅拌2h。再滴加2-甲基四氢-4H-吡喃-4-酮(4.9g,42.9mmol)的无水四氢呋喃溶液(50mL)耗时10min,然后自然升至室温，反应16h。加入饱和氯化铵(200mL)淬灭反应，用乙酸乙酯(200mLx3)萃取，饱和食盐水(200mL)洗涤，无水硫酸钠干燥，浓缩后正相纯化(石油醚：乙酸乙酯＝10:1)得到标题产物(4.9g，黄色油状物)产率80％。1H NMR(400MHz,CDCl3)5.95–5.76(m,1H),4.07–3.95(m,1H),3.54(m,3H),3.43–3.26(m,2H),2.62(m,1H),2.01–1.78(m,2H),1.70–1.55(m,1H),1.20(m,3H).
5-2：2-甲基四氢-2H-吡喃-4-甲醛
将4-(甲氧基亚甲基)-2-甲基四氢-2H-吡喃(4.9g,34.4mmol)溶于甲酸(59mL)中，并加入水(30mL),加热到90℃反应6h。冷却后加入水(300mL)，搅拌30min，并用6N NaOH调节pH＝8～9，用二氯甲烷(150ml×3)萃取，饱和食盐水(100mL)洗涤，无水硫酸钠干燥，浓缩后正相纯化(石油醚：乙酸乙酯＝2:1)得到标题产物(0.7g，黄色油状物)产率16％。1H NMR(400MHz,CDCl3)9.67(s,1H),4.09(m,1H),3.60–3.36(m,2H),2.51(m,1H),1.84(m,2H),1.56(m,1H),1.24(d,J＝6.2Hz,3H),1.22–1.14(m,1H).
5-3：7-碘-8-甲氧基-3-(2-甲基四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶
将2-甲基四氢-2H-吡喃-4-甲醛(400mg，1.51mmol)，2-肼基-4-碘-3-甲氧基吡啶(193mg,1.51 mmol)溶入到乙醇(8ml)中并将此反应于氮气保护下加热到90℃反应2h。将反应液浓缩，溶于二氯甲烷(16mL)，加入碘(460mg,1.81mmol)，碳酸钾(625mg,4.53mmol)，并于室温下搅拌16h。加入水(100mL)，用二氯甲烷(50mL×3)萃取，饱和硫代硫酸钠(50mL)洗涤，饱和食盐水(50mL)洗涤，无水硫酸钠干燥，浓缩后反相纯化(0.5％HCOOH～CH3CN)后得到标题产物(241mg，白色固体)产率43％。LCMS:m/z[M+H]+＝374
化合物5：7-乙基-4-(4-氟-3-(8-甲氧基-3-(2-甲基四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶 -7-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-碘-8-甲氧基-3-(2-甲基四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶(100mg,0.27mmol)，(2-氟-5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)苯基)硼酸(77mg,0.27mmol)为原料，得到标题化合物(35mg,白色固体)产率27.0％。
1H NMR(400MHz,DMSO-d6)9.58(s,1H),8.88(s,1H),8.68–8.49(m,2H),8.44(d,1H),7.66–7.53(m,1H),7.04(d,1H),4.52(q,2H),4.31(s,3H),4.02(m,1H),3.61(m,3H),2.01(m,2H),1.84(m,1H),1.65–1.46(m,4H),1.18(d,3H).LCMS:m/z[M+H]+＝488
实施例6
化合物6：7-乙基-4-(4-氟-3-(8-甲氧基-3-(3-甲氧基环丁基)-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)- 7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-碘-8-甲氧基-3-(3-甲氧基环丁基)-[1,2,4]三唑并[4,3-a]吡啶(85mg,0.24mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(66mg,0.24mmol)为原料，得到标题化合物(41.0mg，白色固体)产率36.2％。
1H NMR(400MHz,DMSO-d6)9.58(s,1H),8.88(s,1H),8.63–8.46(m,2H),8.18(d,J＝7.0Hz,1H),7.59(t,J＝9.2Hz,1H),7.03(d,J＝7.0Hz,1H),4.52(q,J＝7.2Hz,2H),4.32(s,3H),4.02(m,1H),3.61(m,1H),3.21(s,3H),2.87(m,2H),2.31(m,2H),1.56(t,J＝7.3Hz,3H).LCMS:m/z[M+1]+＝474
实施例7
7-1：4-氯-2-(氯甲基)-3-甲氧基吡啶
4-氯-3-甲氧基-2-甲基吡啶1-氧化物(4.9g,28.31mmol)溶于无水二氯甲烷(250mL)中,并于氮气保护下同时缓慢滴加三氯氧磷(6.5g，42.47mmol)溶解于二氯甲烷(40mL)和三乙胺(5.7g,56.62mmol)溶解于二氯甲烷(40mL)，并保持同一速度，同时加完后，于30℃搅拌3h。将反应液倒入饱和碳酸氢钠溶液300mL中，用二氯甲烷(200ml×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩得到标题产物(4.9g，黄色固体)产率89％。LCMS:m/z[M+1]+＝192
7-2：2–((4-氯-3-甲氧基吡啶-2-基)甲基)异吲哚-1,3-二酮
将4-氯-2-(氯甲基)-3-甲氧基吡啶(5.4g，28.31mmol),邻苯二甲酰亚胺钾盐(5.2g,28.31mmol)溶入N，N-二甲基甲酰胺(100.0ml)中，并将此反应于氮气保护下于60℃反应16h。将反应液倒入水(100mL)中。用乙酸乙酯(100mlx3)萃取，分别用5％氯化锂和饱和食盐水洗涤，无水硫酸钠干燥，浓缩后得到一粗品，正相纯化(CH2Cl2～乙酸乙酯)后得到标题产物(3.7g，白色固体)产率48％。1H NMR(400MHz,CDCl3)8.09(d,J＝5.2Hz,1H),7.91–7.85(m,2H),7.78–7.71(m,2H),7.21(d,J＝5.2Hz,1H),5.08(s,2H),4.03(s,3H).LCMS:m/z[M+1]+＝303
7-3：(4-氯-3-甲氧基吡啶-2-基)甲胺
将2–((4-氯-3-甲氧基吡啶-2-基)甲基)异吲哚-1,3-二酮(3.8g，12.53mmol)溶入乙醇(38ml)中，并加入水和肼(7.6mL)并将此反应于80℃反应16h。将反应液旋去乙醇后，加入50mL水，并用浓盐酸调节pH＝1，有大量固体析出，过滤，收集滤液，并用NaOH调节PH＝9～10后用乙酸乙酯(100mlx3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩后得到标题产物(1.35g，黄色油状物)产率62％。LCMS:m/z[M+1]+＝173
7-4：N-((4-氯-3-甲氧基吡啶-2-基)甲基)四氢-2H-吡喃-4-甲酰胺
将(4-氯-3-甲氧基吡啶-2-基)甲胺(300mg，1.74mmol)溶于二氯甲烷(6mL)中，并加入二异丙基乙基胺(562mg,4.35mmol),然后将4-氢吡喃-4-甲酰氯(258.5mg,1.74mmol)缓慢滴加入反应液，并将此反应于30℃反应3h。将反应液倒入50mL水中，并用二氯甲烷(50mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩后得到标题产物(497mg，黄色油状物)。
7-5：7-氯-8-甲氧基-3-(四氢-2H-吡喃-4-基)咪唑并[1,5-a]吡啶
将N-((4-氯-3-甲氧基吡啶-2-基)甲基)四氢-2H-吡喃-4-甲酰胺(200mg，0.7mmol)溶入到无水四氢呋喃(4ml)中，并加入Burgess试剂(502mg,2.1mmol)，并将此反应于N2保护下于60℃反应3h。将反应液倒入水(50mL)中。用乙酸乙酯(50ml×3)萃取，浓缩后得到一粗品，反相纯化(0.5％HCOOH～CH3CN)后得到标题产物(102mg，白色固体)产率54％。LCMS:m/z[M+1]+＝267
7：7-乙基-4-(4-氟-3-(8-甲氧基-3-(四氢-2H-吡喃-4-基)咪唑并[1,5-a]吡啶-7-基)苯基)-7H-咪唑 并[4,5-c]哒嗪
实验操作同实施例2，以7-氯-8-甲氧基-3-(四氢-2H-吡喃-4-基)咪唑并[1,5-a]吡啶(102.1mg,0.38mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(109.5mg,0.38mmol)为原料，得到标题化合物(22mg，白色固体)产率12％。
1H NMR(400MHz,DMSO-d6)9.57(s,1H),8.87(s,1H),8.56(d,1H),8.52–8.38(m,1H),8.19(d,1H),7.61(s,1H),7.55(t,1H),6.71(d,1H),4.52(q,J＝7.2Hz,2H),4.04–3.88(m,5H),3.53(m,3H),1.96–1.75(m,4H),1.56(t,J＝7.2Hz,3H).LCMS:m/z[M+1]+＝473
实施例8

8-1：5-氨基-1-甲基-1H-吡唑-4-甲醛
将5-氨基-1-甲基-1H-吡唑-4-甲腈(5g，128mmol)加入四氢呋喃(20ml)中，氩气保护下，降温至-20℃，滴加二异丁基氢化铝(40mL，40mmol)，加完后室温搅拌16小时，将反应液倒入冰水(50mL)中，过滤，滤液用乙酸乙酯萃取(50mL×3)，浓缩，柱层析纯化(二氯甲烷:甲醇＝15:1),得到白色固体化合物(420mg,产率20％)。LC-MS:m/z[M+H]+＝126.0
8-2：3-(5-氨基-1-甲基-1H-吡唑-4-基)丙烯酸甲酯
将18-冠-6(2.9g，11.0mmol)用四氢呋喃(20mL)溶解后，降温到-40℃,再加入2-(二甲氧基磷酰基)乙酸甲酯(420mg，2.2mmol)，双(三甲基硅基)胺基锂(2.2mL，2.2mmol)，将5-氨基-1-甲基-1H-吡唑-4-甲醛(275mg，2.2mmol)用四氢呋喃(10ml)溶解后，滴加到体系中，然后室温搅拌16小时，将反应液倒入水(50mL)中，乙酸乙酯萃取(50mL×3)，浓缩，柱层析纯化(石油醚:乙酸乙酯＝1:1),得到白色固体化合物(200mg,产率50％)LC-MS:m/z[M+H]+＝182
8-3：1-甲基-1H-吡唑并[3,4-b]吡啶-6-醇
将3-(5-氨基-1-甲基-1H-吡唑-4-基)丙烯酸甲酯(200mg，1.1mmol)用稀盐酸(3mol/L,20mL)溶解后，100℃搅拌4小时。直接旋干，再用蒸馏水(5mL)加热溶解，静置在冰箱上层冷却，大量白色絮状沉淀析出，过滤，得到白色固体(150mg,产率91％)。LC-MS:m/z[M+H]+＝150
8-4：5-溴-1-甲基-1H-吡唑并[3,4-b]吡啶-6-醇
将1-甲基-1H-吡唑并[3,4-b]吡啶-6-醇(150mg，1.0mmol)加入到乙酸(15mL)中，滴加液溴(480mg，3.0mmol)，之后室温搅拌2h，反应液直接旋干，加入去离子水(20mL)，大量白色固体析出，过滤，得到白色固体(100mg,产率44％)。LC-MS:m/z[M+H]+＝228
8-5：5-溴-6-甲氧基-1-甲基-1H-吡唑并[3,4-b]吡啶
将5-溴-1-甲基-1H-吡唑并[3,4-b]吡啶-6-醇(114mg，0.5mmol)加入到N，N-二甲基甲酰胺(20mL)中，加入碳酸铯(326mg，1.0mmol)和碘甲烷(142mg,1.0mmol)，60℃反应2小时，将反应液倒入冰水(50mL)中，乙酸乙酯萃取(50mLx3)，浓缩，柱层析纯化(石油醚:乙酸乙酯＝8:1),得到白色固体化合物(120mg,产率99％)。LC-MS:m/z[M+H]+＝242
8：7-乙基-4-(4-氟-3-(6-甲氧基-1-甲基-1H-吡唑并[3,4-b]吡啶-5-基)苯基)-7H-咪唑并[4,5-c]哒嗪实验操作同实施2，以5-溴-6-甲氧基-1-甲基-1H-吡唑并[3,4-b]吡啶(100mg，0.40mmol)，7-乙基-4-(4-氟-3-(4，4，5，5-四甲基-1，3，2-二氧杂硼烷-2-基)苯基-7H-咪唑并[4,5-c]哒嗪(125mg,0.44mmol)为原料，得到白色固体(30mg，收率：20％)。
1H NMR(400MHz,CDCl3)9.42(s,1H),8.36(s,1H),8.30(s,2H),7.96(s,1H),7.92(s,1H),7.37(t,1H),4.59(d,J＝6.8Hz,2H),4.11(s,3H),4.06(s,3H),1.70(t,J＝6.4Hz,3H).LC-MS:m/z[M+H]+＝404
实施例9
9-1：6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-羧酸甲酯
5-溴-4-甲氧基吡啶-2-胺(1g,3.5mmol)和3-溴-2-氧代丙酸甲酯(762mg，4.2mmol)加到无水乙醇中(10mL)闷罐90℃搅拌过夜。将反应液直接上制备板分离纯化(石油醚/乙酸乙酯＝3/1)得标题化合物(600mg,60％)。
9-2：6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-羧酸
6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-羧酸甲酯(400mg,1.4mmol)和(2M)氢氧化钠水溶液(2mL)加到四氢呋喃中(4mL)室温搅拌过夜。将反应液浓缩，用乙酸乙酯萃取水相三次，有机相浓缩上制备板分离纯化(二氯甲烷/甲醇＝20/1)得标题化合物(300mg,79％)。
9-3：(6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-基)(吡咯烷-1-基)甲酮
6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-羧酸(300mg,1.1mmol),1-羟基苯并三唑(234mg,1.1mmol),1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(95mg,0.5mmol),N,N-二异丙基乙胺(425mg，3.3mmol)和吡咯烷(213mg,3.3mmol)加到N，N-二甲基甲酰胺(6mL)35℃搅拌过夜。反应直接上制备板分离纯化(二氯甲烷/甲醇＝20/1)得标题化合物(60mg，17％)。
9：(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑并[1,2-a]吡啶-2-基)(吡咯 烷-1-基)甲酮
实验操作同实施例2，以(6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-基)(吡咯烷-1-基)甲酮(60mg,0.19mmol)和7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(72mg,0.19mmol)为原料，得标题化合物(50mg，54％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H),8.30(br.s.,1H),8.15-8.04(m,1H),7.79-7.67(m,1H),7.53(br.s.,1H),7.38(br.s.,1H),4.59(d,J＝6.8Hz,1H),4.13(br.s.,1H),3.90(br.s.,3H),3.75(br.s.,1H),2.03-1.93(m,4H),1.70(d,J＝7.3Hz,1H).LC-MS:m/z[M+H]+＝486.
实施例10
10-1：1-(乙基磺酰基)-1,2,3,4-四氢吡啶并[2,3-b]吡嗪
将1,2,3,4-四氢吡啶并[2,3-b]吡嗪(500mg,3.7mmol)，溶于四氢呋喃(10mL)中，加入三乙胺(1.12g,11.1mmol)，再加入乙磺酰氯(480mg，3.7mmol)，室温搅拌12小时，用乙酸乙酯(200mL)稀释后，用水洗(150mLx2)后，干燥，浓缩，柱层析纯化(石油醚:乙酸乙酯＝1:1)后得到黄色固体(300mg,收率35.7％)。LC-MS:m/z[M+H]+＝228.0
10：4-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-1-(乙基磺酰基)-1,2,3,4-四氢吡啶并[2, 3-b]吡嗪
将1-(乙基磺酰基)-1,2,3,4-四氢吡啶并[2,3-b]吡嗪(200mg,0.88mmol)，4-(3-氯-4-氟苯基)-7-乙基-7H-咪唑并[4,5-c]哒嗪(364mg,1.32mmol)，磷酸钾(374mg,1.76mmol)，三(二苯亚甲基丙 酮)二钯(80mg,0.09mmol)，和甲苯(20mL)依次加入到反应瓶中，加热到90℃反应16小时。反应液直接浓缩，用prep-HPLC(0.5％NH3.H2O～乙腈)纯化得到白色固体(30mg，收率15％)。
1H NMR(400MHz,CDCl3)9.39(s,1H),8.32(m,,2H),8.28–8.19(m,1H),7.91(d,J＝4.8Hz,1H),7.85(d,J＝8.0Hz,1H),7.43–7.33(m,1H),6.74(m,1H),4.58(q,J＝7.3Hz,2H),4.14–4.00(m,2H),4.00–3.90(m,2H),3.18(q,J＝7.4Hz,2H),1.70(t,J＝7.3Hz,3H),1.44(t,J＝7.4Hz,3H).LC-MS:m/z[M+H]+＝468
实施例11
11-1：4-溴-2-硝基吡啶-3-醇
在氮气的惰性气氛下，将4-溴吡啶-3-醇(3g，17.3mmol)加入到8mL的浓硫酸中搅拌15分钟。在0℃下滴加发烟硝酸(1.64g，26mmol)搅拌过夜。反应液10g冰中，用乙酸乙酯(10mLx3)萃取，旋干后得到粗品，正相硅胶柱层析(石油醚：乙酸乙酯＝10:1～1:1)纯化后，得到标题产物(2.19g，产率56％)黄色固体。LC-MS:m/z[M+H]+＝219
11-2：4-溴-3-甲氧基-2-硝基吡啶
将4-溴-2-硝基吡啶-3-醇(2.19g,10mmol)，碳酸钾(1.39g,20mmol)溶于N，N-二甲基甲酰胺(17.0mL)中，滴加碘甲烷(1.43g,20mmol)，室温搅拌17小时，用水(20mL)淬灭后，再用乙酸乙酯(20mLx2)萃取,旋干后得到粗品，正相硅胶柱层析(石油醚：乙酸乙酯＝10:1～2:1)纯化后，得到标题产物(0.65g，产率28％)白色固体。LC-MS:m/z[M+H]+＝233
11-3：4-溴-3-甲氧基吡啶-2-胺
将4-溴-3-甲氧基-2-硝基吡啶(550mg,2.37mmol)和氯化铵(628mg,11.85mmol)溶于乙醇:水＝1:1(16.6mL)中，加热到60℃，分批加入铁粉(664mg，11.85mmol)，反应在60℃继续搅拌2小时，过滤，滤饼用甲醇(10mLx3)淋洗，将淋洗液合并旋干。用5mL水稀释，水相再用乙酸乙酯(10mLx3)萃取,旋干后得到标题产物(514mg，100％)棕色固体。LC-MS:m/z[M+H]+＝203
11-4：7-溴-8-甲氧基咪唑并[1,2-a]吡啶
将4-溴-3-甲氧基吡啶-2-胺(200mg,0.99mmol)，氯乙醛(295mg,40Wt％，1.5mmol)，碳酸氢钠(159mg,1.5mmol)溶于乙醇(10.0mL)的溶液中，在微波反应器中加热到90℃反应1小时,。将反应液浓缩，用2mL水稀释，水相再用二氯甲烷(2mLx3)萃取，有机相合并，干燥旋干，得到标题产物(290mg，100％)棕色固体。LC-MS:m/z[M+H]+＝227
11-5：7-溴-3-碘-8-甲氧基咪唑并[1,2-a]吡啶
将7-溴-8-甲氧基咪唑并[1,2-a]吡啶(400mg,1.28mmol)溶于N，N-二甲基甲酰胺(7.5mL)中，加入N-碘代丁二酰亚胺(375mg，1.67mmol)，反应在100℃搅拌1小时。冷却至室温，用10mL水稀释，水相再用甲基叔丁基醚(10mLx3)萃取,旋干后得到标题产物(220mg，100％)棕色固体。LC-MS:m/z[M+H]+＝353
11-6：7-溴-3-环丙基-8-甲氧基咪唑并[1,2-a]吡啶
将7-溴-3-碘-8-甲氧基咪唑并[1,2-a]吡啶(400mg,1.13mmol)，环丙基硼酸(97mg,1.13mmol)，四三苯基磷钯(131mg,0.113mmol)，磷酸钾(481mg,2.27mmol)，溶于二氧六环:H2O＝10:1(11mL)中，置换氩气三次，反应在95℃搅拌17小时。冷却至室温，正相硅胶柱层析(石油醚：乙酸乙酯＝10:1～1:1)纯化后，得到标题产物(100mg，33％)黄色固体。LC-MS:m/z[M+H]+＝267
11：4-(3-(3-环丙基-8-甲氧基咪唑并[1,2-a]吡啶-7-基)-4-氟苯基)-7-乙基-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-溴-3-环丙基-8-甲氧基咪唑并[1,2-a]吡啶(100mg，0.37mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(107mg,0.37mmol)为原料,得标题化合物(60mg，37％)类白色固体。
1H NMR(400MHz,CDCl3)9.37(s,1H),8.38–8.30(m,1H),8.29–8.20(m,2H),7.85(d,1H),7.36(dd,2H),6.78(d,1H),4.58(q,2H),4.24(s,3H),2.12(s,1H),1.69(t,3H),0.97(m,4H).LC-MS:m/z[M+H]+＝429
实施例12
12：7-乙基-4-(4-氟-3-(2-(1-氟环丙基)-7-甲氧基咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑并[4,5-c] 哒嗪
实验操作同实施例2，以6-溴-2-(1-氟环丙基)-7-甲氧基咪唑并[1,2-a]吡啶(100.0mg,0.36mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(100mg,0.36mmol)为原料，得到标题产物(30mg，产率17％)白色固体。
1H NMR(400MHz,DMSO-d6)9.38(s,1H),8.29(m,3H),8.05(s,1H),7.55(s,1H),7.37(t,1H),6.99(s,1H),4.59(q,2H),3.85(s,3H),1.70(t,3H),1.56–1.33(m,4H).LC-MS:m/z[M+H]+＝447
实施例13
13-1：3-溴-1-甲基-1H-吡咯并[3,2-b]吡啶-6-羧酸甲酯
将3-溴-1H-吡咯并[3,2-b]吡啶-6-羧酸甲酯(102mg,0.4mmol)用N，N-二甲基甲酰胺(5mL)溶解后，0℃下加入氰化钠(32mg，0.8mmol)搅拌30min后加入碘甲烷(65mg，0.6mmol)，室温搅拌16小时。将反应液倒入冰水(30mL)中，用乙酸乙酯(50mLx3)萃取，合并有机相，干燥，浓缩，柱层析纯化(石油醚:乙酸乙酯＝3:1)后得到白色固体(100mg,产率92.9％)。LC-MS:m/z[M+H]+＝269
13-2：3-溴-N,1-二甲基-1H-吡咯并[3,2-b]吡啶-6-甲酰胺
将3-溴-1-甲基-1H-吡咯并[3,2-b]吡啶-6-羧酸甲酯(110mg，0.41mmol)溶入甲胺的乙醇溶液(5mL)，80℃搅拌16小时后，直接浓缩，得到白色固体(80mg，产率73.1％)。LC-MS:m/z[M+H]+＝268
13：3-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-N,1-二甲基-1H-吡咯并[3,2-b]吡啶-6- 甲酰胺
实验操作同实施例2，以3-溴-N,1-二甲基-1H-吡咯并[3,2-b]吡啶-6-甲酰胺(80mg,0.30mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(101mg,0.36mmol)为原料，得到标题化合物(45.0mg，白色固体)产率35％.
1H NMR(400MHz,CDCl3)9.59–9.42(m,1H),9.41–9.25(m,1H),8.99–8.88(m,1H),8.30–8.18(m,2H),8.13–8.03(m,1H),8.00–7.92(m,1H),7.35–7.28(m,1H),4.64–4.45(m,2H),4.02–3.89(m,3H),3.15–2.92(m,3H),1.69–1.60(m,3H).LC-MS:m/z[M+H]+＝430
实施例14
14-1：7-碘-8-甲氧基-3-(四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(100mg,0.38mmol)和四氢-2H-吡喃-4-甲醛(50mg,0.38mmol)为原料，得到白色固体标题化合物(100mg,74％)。LC-MS:m/z[M+H]+＝360.
14：4-(4-氟-3-(8-甲氧基-3-(四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7- 异丙基-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)苯基)-7-异丙基-7H-咪唑并[4,5-c]哒嗪(45mg,0.11mmol)和7-碘-8-甲氧基-3-(四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.14mmol)为原料，得标题化合物(40mg，74.5％)。
1H NMR(400MHz,CHLOROFORM-d)9.37(s,1H),8.37-8.31(m,2H),8.27(dd,J＝3.2,5.6Hz,1H),7.74(d,J＝6.8Hz,1H),7.40(t,J＝9.0Hz,1H),6.90(d,J＝6.8Hz,1H),5.30-5.17(m,1H),4.50(s,3H),4.19(d,J＝11.7Hz,2H),3.67(t,J＝10.5Hz,2H),3.43-3.31(m,1H),2.33-2.22(m,2H),2.07(d,J＝13.7Hz,2H),1.77(d,J＝6.8Hz,6H).LC-MS:m/z[M+H]+＝488.
实施例15
15：4,4'-(4-氟-1,3-亚苯基)双(7-乙基-7H-咪唑并[4,5-c]哒嗪)
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(100mg,0.3mmol)，4-氯-7-乙基-7H-咪唑并[4,5-c]哒嗪(5mg,0.3mmol)为原料，得到白色固体标题化合物(17mg,16％)。
1H NMR(400MHz,CHLOROFORM-d)9.45(br.s.,2H)8.84-9.02(m,1H)8.37-8.49(m,1H)8.30(br.s.,2H)7.50(br.s.,1H)4.49-4.69(m,4H)1.71(q,J＝7.01Hz,6H).LC-MS:m/z[M+H]+＝389.
实施例16
16-1：3-(2,2-二甲基四氢-2H-吡喃-4-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
实验从操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(300mg,1.132mmol)，2,2-二甲基四氢-2H-吡喃-4-甲醛(161mg,1.132mmol)为原料，得标题化合物棕色固体(259mg,59％)。LC-MS:m/z[M+H]+＝388.
16：4-(3-(3-(2,2-二甲基四氢-2H-吡喃-4-基)-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)-4-氟苯 基)-7-乙基-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H- 咪唑并[4,5-c]哒嗪(50mg,0.13mmol)，3-(2,2-二甲基四氢-2H-吡喃-4-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(59mg,0.15mmol)为原料，得标题化合物黄色固体(34mg,53％)。
1H NMR(400MHz,CHLOROFORM-d)9.35(s,1H),8.32(dd,J＝2.2,7.1Hz,1H),8.29-8.23(m,2H),7.74(d,J＝7.3Hz,1H),7.37(t,J＝9.0Hz,1H),6.89(d,J＝7.3Hz,1H),4.57(q,J＝7.3Hz,2H),4.46(s,3H),3.97-3.84(m,2H),3.53-3.43(m,1H),2.16-2.07(m,2H),1.99-1.92(m,2H),1.67(t,J＝7.3Hz,3H),1.39(s,3H),1.32(s,3H).LC-MS:m/z[M+H]+＝502.
实施例17
17-1：3-环丁基-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
实验从操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(300mg,1.132mmol)，环丁基甲醛(95mg,1.132mmol)为原料，得标题化合物棕色固体(262mg,70％)。LC-MS:m/z[M+H]+＝330.
17：4-(3-(3-环丁基-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)-4-氟苯基)-7-乙基-7H-咪唑并 [4],5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(60mg,0.152mmol)，3-环丁基-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.152mmol)为原料，得标题化合物黄色固体(34mg,50％)。
1H NMR(400MHz,CHLOROFORM-d)d＝9.37(s,1H),8.35-8.31(m,1H),8.30-8.23(m,2H),7.58(d,J＝6.8Hz,1H),7.38(t,J＝9.0Hz,1H),6.85(d,J＝7.3Hz,1H),4.62-4.55(m,2H),4.48(s,3H),3.88(quin,J＝8.4Hz,1H),2.76-2.54(m,4H),2.29-2.09(m,2H),1.68(t,J＝7.3Hz,3H).LC-MS:m/z[M+H]+＝444.
实施例18
18-1：7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-3(2H)-酮
2-肼基-4-碘-3-甲氧基吡啶(450mg,1.698mmol)，羰基二咪唑(550mg,3.400mmol)加到乙腈(4ml)中，85℃反应2小时。反应液抽滤，滤得固体水洗，抽滤得标题化合物(370mg,75％)。LC-MS:m/z[M+H]+＝292.
18-2：7-碘-8-甲氧基-2-甲基-[1,2,4]三唑并[4,3-a]吡啶-3(2H)-酮
7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-3(2H)-酮(60mg,0.206mmol)，碘甲烷(32mg, 0.227mmol)，碳酸铯(134mg,0.412mmol)加到乙腈(1ml)中，室温搅拌30分钟。反应液抽滤，浓缩，制备薄层色谱(石油醚/乙酸乙酯＝1/1)得标题化合物白色固体(39mg,62％)。LC-MS:m/z[M+H]+＝306.
18：7-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-8-甲氧基-2-甲基-[1,2,4]三唑并[4,3-a] 吡啶-3(2H)-酮
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(42mg,0.115mmol)，3-(4-氟-1-甲基哌啶-4-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(35mg,0.115mmol)为原料，得标题化合物黄色固体(13mg,27％)。
1H NMR(400MHz,CHLOROFORM-d)9.36(s,1H),8.32-8.25(m,3H),7.62(d,J＝6.8Hz,1H),7.38(t,J＝9.0Hz,1H),6.56(d,J＝6.8Hz,1H),4.59(q,J＝7.2Hz,2H),4.14(s,3H),3.73(s,3H),1.69(t,J＝7.1Hz,3H).LC-MS:m/z[M+H]+＝420.
实施例19
19：(7-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-6-甲氧基咪唑并[1,5-a]吡啶-3-基)(吗啉) 甲酮
将7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(60mg,0.15mmol)，(7-溴-6-甲氧基咪唑并[1,5-a]吡啶-3-基)(吗啉)甲酮(50mg,0.15mmol)，[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(CAS:95464-05-4,12mg,0.02mmol)和碳酸铯(200mg,0.3mmol)依次加入到5mL的二氧六环和0.5mL的水中，然后95℃搅拌2小时。过滤反应液，浓缩，制备板纯化(二氯甲烷：甲醇＝20：1)，得到白色固体标题化合物(24mg,33％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H)9.06(s,1H)8.29(s,3H)7.51-7.65(m,2H)7.37(t,J＝8.80Hz,1H)4.59(q,J＝7.34Hz,4H)3.89(s,3H)3.84(br.s.,6H)1.70(t,J＝7.34Hz,3H).LC-MS:m/z[M+H]+＝502.
实施例20
20-1：N-((4-氯-3-甲氧基吡啶-2-基)甲基)环丙烷甲酰胺
将(4-氯-3-甲氧基吡啶-2-基)甲胺(900mg，5.2mmol)，碳酸钾(2.2g，15.7mmol)用乙腈(20mL)溶解后，冰水浴下加入环丙酰氯(816mg，7.8mmol)，室温搅拌2h，反应液直接旋干，柱层析分离(石油醚:乙酸乙酯＝1:1),得到黄色固体(300mg,产率24％)。LC-MS:m/z[M+H]+＝241
20-2：7-氯-3-环丙基-8-甲氧基咪唑并[1,5-a]吡啶
将N-((4-氯-3-甲氧基吡啶-2-基)甲基)环丙烷甲酰胺(200mg，0.83mmol)，Burgess试剂(500mg，2.1mmol)和二氯甲烷(5mL)加入反应瓶中，室温搅拌2小时后直接旋干，柱层析纯化(石油醚:乙酸乙酯＝1:1),得到黄色油状液体(110mg,产率60％)。LC-MS:m/z[M+H]+＝223
20：4-(3-(3-环丙基-8-甲氧基咪唑并[1,5-a]吡啶-7-基)-4-氟苯基)-7-乙基-7H-咪唑并[4,5-c]哒嗪
将7-氯-3-环丙基-8-甲氧基咪唑并[1,5-a]吡啶(50mg，0.23mmol)，7-乙基-4-(4-氟-3-(4，4，5，5-四甲基-1，3，2-二氧杂硼烷-2-基)苯基-7H-咪唑并[4,5-c]哒嗪(97mg,0.34mmol),[1,1-双(二叔丁基膦)二茂铁]二氯化钯(II)(15mg,0.023mmol),碳酸铯(150mg,0.46mmol)加入1，4-二氧六环/水(5ml/1ml)中。氩气保护下，100℃下微波反应2小时。过滤，滤液旋干送HPLC制备，冻干后得标题产物(26mg，产率27％)。
1H NMR(400MHz,DMSO-d6)9.57(s,1H),8.87(s,1H),8.57(dd,J＝7.2,2.3Hz,1H),8.52–8.36(m,1H),8.20(d,J＝7.2Hz,1H),7.62–7.45(m,2H),6.74(d,J＝7.2Hz,1H),4.52(q,J＝7.2Hz,2H),3.95(s,3H),2.41–2.30(m,1H),1.56(t,J＝7.3Hz,3H),1.15–1.03(m,2H),1.00–0.84(m,2H).LC-MS:m/z[M+H]+＝429
实施例21
7-氯-6-甲氧基-2-甲基咪唑并[1,2-a]吡啶
将4-氯-5-甲氧基吡啶-2-胺(20mg，0.13mmol)加入到乙醇(2mL)中，加入氯丙酮(35mg，0.38mmol)和碳酸钠(40mg,0.38mmol)，微波加热到100℃反应1小时，过滤，浓缩，柱层析纯化得到标题产物(5mg,产率20％)。LC-MS:m/z[M+H]+＝197
7-乙基-4-(4-氟-3-(6-甲氧基-2-甲基咪唑并[1,2-a]吡啶-7-基)苯基)-7H-咪唑并[4,5-c]哒嗪
将7-氯-6-甲氧基-2-甲基咪唑并[1,2-a]吡啶(72mg，0.367mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(105mg,0.367mmol),[1,1-双(二叔丁基膦)二茂铁]二氯化钯(II)(24mg,0.0367mmol),碳酸铯(239mg,0.735mmol)加入1，4-二氧六环/水(7.2mL/0.8mL)中。氩气保护下，95℃反应2小时。加入10mL水，用乙酸乙酯(20mLx2)萃取，有机相用无水硫酸钠干燥，浓缩，用prep-HPLC(0.5％NH3.H2O～乙腈)纯化得到白色固体(40mg，收率：31％)。
1H NMR(400MHz,CDCl3)9.38(s,1H),8.40–8.16(m,3H),7.71(s,2H),7.43–7.29(m,2H),4.58(q,J＝7.3Hz,2H),3.84(s,3H),2.51(s,3H),1.69(t,J＝7.3Hz,3H).LC-MS:m/z[M+H]+＝403.
实施例22
22-1：3-(4-氟哌啶-4-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
2-肼基-4-碘-3-甲氧基吡啶(300mg,0.822mmol)，4-氟-4-甲酰基哌啶-1-羧酸叔丁酯(285mg,1.233mmol)加到乙醇(2ml)中，25℃搅拌2小时。溴化铜(37mg,0.164mmol),过硫酸氢钾(606mg, 0.986mmol)加入反应液，25℃搅拌1.5小时。反应液抽滤，浓缩，溶于二氯甲烷(5ml)中，盐酸-乙酸乙酯溶液(4mol/L,2ml)加入反应液，25℃搅拌1小时。反应液浓缩，蒸干，制备薄层色谱(二氯甲烷/甲醇＝15/1)得标题化合物黄色固体(179mg,61％)。LC-MS:m/z[M+H]+＝377.
22-2：3-(4-氟-1-甲基哌啶-4-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
3-(4-氟哌啶-4-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(179mg,0.500mmol)，甲醛水溶液(60mg,1mmol)加到四氢呋喃(1ml)中，25℃搅拌5分钟。氰基硼氢化钠(63mg,1mmol)加到反应液，25℃搅拌1小时。反应液抽滤，浓缩，制备薄层色谱(二氯甲烷/甲醇＝20/1)得标题化合物棕色固体(79mg,42％)。LC-MS:m/z[M+H]+＝391.
22：7-乙基-4-(4-氟-3-(3-(4-氟-1-甲基哌啶-4-基)-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯 基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(71mg,0.195mmol)，3-(4-氟-1-甲基哌啶-4-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(76mg,0.195mmol)为原料，得标题化合物淡黄色固体(6mg,6％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H),8.36(d,J＝6.8Hz,1H),8.28(s,2H),8.15(d,J＝6.8Hz,1H),7.41(t,J＝9.0Hz,1H),6.93(d,J＝7.3Hz,1H),4.59(q,J＝7.3Hz,2H),4.48(s,3H),3.04(br.s.,2H),2.88-2.57(m,6H),2.53(s,3H),1.70(br.s.,3H).LC-MS:m/z[M+H]+＝505.
实施例23
23-1：3-(氮杂环丁烷-3-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例22中，22-1的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(250mg,0.94mmol)，3-甲酰基氮杂环丁烷-1-羧酸叔丁酯(349mg,1.89mmol)为原料，得标题化合物棕色固体(136mg,44％)。LC-MS:m/z[M+H]+＝331.
23-2：7-碘-8-甲氧基-3-(1-(2,2,2-三氟乙基)氮杂环丁烷-3-基)-[1,2,4]三唑并[4,3-a]吡啶
3-(氮杂环丁烷-3-基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(136mg,0.41mmol)，2,2,2-三氟乙基三氟甲磺酸酯(113mg,0.50mmol)，碳酸钾(114mg,0.82mmol)加到乙腈(3ml)中，50℃反应2小时。反应液抽滤，浓缩，制备薄层色谱(二氯甲烷/甲醇＝30/1)得标题化合物黄色胶状固体(70mg,41％)。LC-MS:m/z[M+H]+＝413.
23：7-乙基-4-(4-氟-3-(8-甲氧基-3-(1-(2,2,2-三氟乙基)氮杂环丁烷-3-基)-[1,2,4]三唑并[4,3-a]吡 啶-7-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(62mg,0.17mmol)，7-碘-8-甲氧基-3-(1-(2,2,2-三氟乙基)氮杂环丁烷-3-基)-[1,2,4]三唑并[4,3-a]吡啶(70mg,0.17mmol)为原料，得标题化合物淡黄色固体(22mg,25％)。
1H NMR(400MHz,CHLOROFORM-d)9.36(s,1H),8.34(d,J＝5.9Hz,1H),8.28(s,2H),7.98(d,J＝6.8Hz,1H),7.39(t,J＝9.0Hz,1H),6.93(d,J＝6.8Hz,1H),4.58(q,J＝7.3Hz,2H),4.46(s,3H),4.27(quin,J＝7.5Hz,1H),4.09(t,J＝7.6Hz,2H),3.88(t,J＝7.1Hz,2H),3.17(q,J＝9.3Hz,2H),1.68(t,J＝7.3Hz,3H).LC-MS:m/z[M+H]+＝527.
实施例24
24-1：7-碘-8-甲氧基-3-(4-甲基四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(100mg,0.38mmol)，4-甲基四氢-2H-吡喃-4-甲醛(96mg,0.75mmol)为原料，得标题化合物棕红色固体(113mg,80％)。LC-MS:m/z[M+H]+＝374.
24：7-乙基-4-(4-氟-3-(8-甲氧基-3-(4-甲基四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶-7-基) 苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(54mg,0.15mmol)，7-碘-8-甲氧基-3-(4-甲基四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶(55mg,0.15mmol)为原料，得标题化合物黄色固体(19mg,27％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H),8.35(d,J＝5.9Hz,1H),8.28(s,2H),7.94(d,J＝7.3Hz,1H),7.40(t,J＝9.0Hz,1H),6.87(d,J＝6.8Hz,1H),4.59(q,J＝7.3Hz,2H),4.49(s,3H),3.93-3.85(m,2H),3.83-3.73(m,2H),2.60(d,J＝14.2Hz,2H),2.00-1.91(m,2H),1.69(br.s.,3H),1.59(s,3H).LC-MS:m/z[M+H]+＝488.
实施例25
25-1：3-(4,4-二氟环己基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(100mg,0.38mmol)，4,4-二氟环己烷-1-甲醛(112mg,0.75mmol)为原料，得标题化合物棕红色固体(100mg,67％)。LC-MS:m/z[M+H]+＝394.
25：4-(3-(3-(4,4-二氟环己基)-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)-4-氟苯基)-7-乙基-7H- 咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(52mg,0.14mmol)，3-(4,4-二氟环己基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(55mg,0.14mmol)为原料，得标题化合物淡黄色固体(13mg,20％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(br.s.,1H),8.34(dd,J＝2.0,6.8Hz,1H),8.30-8.25 (m,2H),7.71(d,J＝6.8Hz,1H),7.39(t,J＝9.0Hz,1H),6.92(d,J＝7.3Hz,1H),4.61-4.56(m,2H),4.49(s,3H),3.25(br.s.,1H),2.39(d,J＝9.3Hz,2H),2.29-2.20(m,4H),1.88-1.78(m,2H),1.69(t,J＝7.3Hz,3H).LC-MS:m/z[M+H]+＝508.
实施例26
26-1：6-溴-7-甲氧基-2-甲基咪唑并[1,2-a]吡啶
5-溴-4-甲氧基吡啶-2-胺(200mg,1mmol)和溴丙酮(274mg,2mmol)加到无水EtOH(5mL)90℃搅拌过夜，反应液浓缩后，制备板分离纯化(石油醚/乙酸乙酯＝1/1)得标题化合物(120mg,50％)。
26：4-(4-氟-3-(7-甲氧基-2-甲基咪唑并[1,2-a]吡啶-6-基)苯基)-7-异丙基-7H-咪唑并[4,5-c] 哒嗪
实验操作同实施例2，以6-溴-7-甲氧基-2-甲基咪唑并[1,2-a]吡啶(24.2mg,0.1mmo)和7-异丙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(40mg,0.1mmol)为原料，得标题化合物(17mg,42％)。
1H NMR(400MHz,DMSO-d6)9.55-9.51(s,1H),8.93-8.89(s,1H),8.64-8.41(m,4H),7.60-7.49(m,2H),5.12-5.06(m,1H),3.87-3.78(m,3H),2.39-2.24(m,3H),1.66(d,J＝6.8Hz,6H).LC-MS:m/z[M+H]+＝417.
实施例27
27-1：N'-(4-碘-3-甲氧基吡啶-2-基)环丙烷碳酰肼
实验操作同实施例3中，3-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(200mg,0.76mmol)，环丙甲酸(65mg,0.76mmol)为原料,得到标题产物(190.1mg，白色固体)产率76％。LC-MS:m/z[M+H]+＝334
27-2：3-环丙基-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例3中，3-4的合成方法，以N'-(4-碘-3-甲氧基吡啶-2-基)环丙烷碳酰肼(150.0mg，0.45mmol)为原料，得到标题产物(60.1mg，白色固体)产率32％。LC-MS:m/z[M+H]+＝316
27：4-(3-(3-环丙基-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)-4-氟苯基)-7-异丙基-7H-咪唑并 [4],5-c]哒嗪
实验操作同实施例2，以3-环丙基-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(100.0mg,0.32mmol)，(2-氟-5-(7-异丙基-7H-咪唑并[4,5-c]哒嗪-4-基)苯基)硼酸(142.8mg,0.48mmol)为原料，得 到标题化合物(62.1mg,白色固体)产率37％。
1H NMR(400MHz,DMSO-d6)9.57(s,1H),8.94(s,1H),8.65–8.48(m,2H),8.40(d,J＝7.0Hz,1H),7.59(t,J＝9.2Hz,1H),7.07(d,J＝7.0Hz,1H),5.13(m,1H),4.31(s,3H),2.44(m,1H),1.68(d,J＝6.7Hz,6H),1.27–0.98(m,4H).LC-MS:m/z[M+H]+＝444
实施例28
28-1：7-碘-8-甲氧基-3-(氧杂环丁烷-3-基)-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(200mg,0.754mmol)，氧杂环丁烷-3-甲醛(207mg,3.77mmol)为原料，得标题化合物棕色固体(151mg,61％)。LC-MS:m/z[M+H]+＝332.
28：7-乙基-4-(4-氟-3-(8-甲氧基-3-(氧杂环丁烷-3-基)-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7H- 咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(56mg,0.151mmol)，7-碘-8-甲氧基-3-(氧杂环丁烷-3-基)-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.151mmol)为原料，得标题化合物黄色固体(26mg,39％)。
1H NMR(400MHz,CHLOROFORM-d)d＝9.38(br.s.,1H),8.39-8.24(m,3H),7.92-7.85(m,1H),7.40(t,J＝9.3Hz,1H),6.96(d,J＝4.9Hz,1H),5.22(d,J＝7.3Hz,4H),4.79(br.s.,1H),4.63-4.55(m,2H),4.49(d,J＝2.9Hz,3H),1.69(br.s.,3H).LC-MS:m/z[M+H]+＝446.
实施例29
29-1：7-碘-8-甲氧基-3-异丙基-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(200mg,0.754mmol)，异丁醛(271mg,3.77mmol)为原料，得标题化合物棕色固体(109mg,46％)。LC-MS:m/z[M+H]+＝318.
29：7-乙基-4-(4-氟-3-(8-甲氧基-3-异丙基-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7H-咪唑并[4,5-c] 哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(58mg,0.158mmol)，7-碘-8-甲氧基-3-异丙基-[1,2,4]三唑并[4,3-a]吡啶(50mg, 0.158mmol)为原料，得标题化合物黄色固体(17mg,25％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(br.s.,1H),8.37-8.23(m,3H),7.72(d,J＝6.4Hz,1H),7.43-7.35(m,1H),6.89(d,J＝6.8Hz,1H),4.59(d,J＝7.3Hz,2H),4.48(br.s.,3H),3.39(d,J＝5.9Hz,1H),1.70(d,J＝7.8Hz,3H),1.57(d,J＝5.9Hz,6H).LC-MS:m/z[M+H]+＝432.
实施例30
30-1：6-溴-2-乙基-7-甲氧基咪唑并[1,2-a]吡啶
将5-溴-4-甲氧基吡啶-2-胺(200mg,0.99mmol)和溴丁酮(747mg,4.95mmol)依次加到无水乙醇(5mL)中，在90℃下搅拌16小时。反应液浓缩，制备薄层色谱(石油醚/丙酮＝4/1)分离得油状液体标题化合物(150mg,60％)。LC-MS:m/z[M+H]+＝255,257。
30：7-乙基-4-(3-(2-乙基-7-甲氧基咪唑并[1,2-a]吡啶-6-基)-4-氟苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以6-溴-2-乙基-7-甲氧基咪唑并[1,2-a]吡啶(50mg,0.20mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(74mg,0.20mmol)为原料，得棕色固体标题化合物(10mg,12％)。
1H NMR(400MHz,CHLOROFORM-d)9.37(s,1H),8.28(br.s.,3H),8.02(br.s.,1H),7.36(t,J＝8.8Hz,1H),7.21(br.s.,1H),7.05(br.s.,1H),4.62-4.55(m,2H),3.87(br.s.,3H),2.82(d,J＝7.8Hz,2H),1.69(t,J＝7.1Hz,3H),1.34(d,J＝10.3Hz,3H).LC-MS:m/z[M+H]+＝417.
实施例31
31-1：7-碘-8-甲氧基-3-乙基-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(200mg,0.75mmol)，丙醛(219mg,3.77mmol)为原料，得标题化合物棕色固体(147mg,64％)。LC-MS:m/z[M+H]+＝304.
31：7-乙基-4-(4-氟-3-(8-甲氧基-3-乙基-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7H-咪唑并[4,5-c] 哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(60mg,0.165mmol)，7-碘-8-甲氧基-3-乙基-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.165mmol)为原料，得标题化合物黄色固体(23mg,33％)。
1H NMR(400MHz,CHLOROFORM-d)9.37(br.s.,1H),8.38-8.22(m,3H),7.68(d,J＝4.4Hz, 1H),7.38(t,J＝9.0Hz,1H),6.89(d,J＝5.9Hz,1H),4.58(d,J＝7.3Hz,2H),4.46(br.s.,3H),3.13(d,J＝7.3Hz,2H),1.72-1.66(m,3H),1.56-1.51(m,3H).LC-MS:m/z[M+H]+＝418.
实施例32
32-1：7-碘-8-甲氧基-3-甲基-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(200mg,0.75mmol)，乙醛-四氢呋喃溶液(0.75ml,3.77mmol)为原料，得标题化合物棕色固体(140mg,64％)。LC-MS:m/z[M+H]+＝290.
32：7-乙基-4-(4-氟-3-(8-甲氧基-3-甲基-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7H-咪唑并[4,5-c] 哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(67mg,0.182mmol)，7-碘-8-甲氧基-3-甲基-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.182mmol)为原料，得标题化合物黄色固体(27mg,37％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H),8.34(d,J＝6.4Hz,1H),8.28(s,2H),7.66(d,J＝6.8Hz,1H),7.39(t,J＝9.0Hz,1H),6.91(d,J＝6.8Hz,1H),4.58(q,J＝7.0Hz,2H),4.47(s,3H),2.79(s,3H),1.69(br.s.,3H).LC-MS:m/z[M+H]+＝404.
实施例33
33-1：7-碘-8-甲氧基-3-(四氢呋喃-3-基)-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(100mg,0.38mmol)和四氢呋喃-3-甲醛(50mg,0.38mmol)为原料，得到白色固体标题化合物(140mg,100％)。LC-MS:m/z[M+H]+＝346.
33：7-乙基-4-(4-氟-3-(8-甲氧基-3-(四氢呋喃-3-基)-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7H-咪 唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(60mg,0.17mmol)，7-碘-8-甲氧基-3-(四氢呋喃-3-基)-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.17mmol)为原料，得到黄色固体标题化合物(47mg,60％)。
1H NMR(400MHz,CHLOROFORM-d)9.39(s,1H)8.35(d,J＝6.36Hz,1H)8.29(s,2H)7.83(d,J＝6.85Hz,1H)7.38-7.43(m,1H)6.93(d,J＝6.85Hz,1H)4.59(q,J＝6.85Hz,2H)4.49(s,3H)4.30(d, J＝8.31Hz,1H)4.23(dd,J＝15.41,7.09Hz,2H)3.91-4.08(m,2H)2.53(d,J＝6.85Hz,2H)1.69-1.72(m,3H).LC-MS:m/z[M+H]+＝460.
实施例34
34：7-乙基-4-(4-氟-3-(8-甲氧基-3-(四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶-7-yl)苯 基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(70mg,0.2mmol)，7-碘-8-甲氧基-3-(四氢-2H-吡喃-4-基)-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.2mmol)为原料，得到棕色固体标题化合物(47mg,50％)。
1H NMR(400MHz,CHLOROFORM-d)9.39(s,1H)8.35(d,J＝5.87Hz,1H)8.29(s,2H)7.75(d,J＝6.85Hz,1H)7.40(t,J＝9.05Hz,1H)6.91(d,J＝6.85Hz,1H)4.59(q,J＝7.34Hz,2H)4.50(s,3H)4.19(d,J＝11.25Hz,2H)3.67(t,J＝11.00Hz,2H)3.37(t,J＝11.00Hz,1H)2.20-2.35(m,2H)2.08(d,J＝13.21Hz,2H)1.68(br.s.,3H).LC-MS:m/z[M+H]+＝474.
实施例35
35-1：3-(二氟甲基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
将2-肼基-4-碘-3-甲氧基吡啶(160mg,0.57mmol)溶于2mL二氟醋酸酐中，加热回流过夜。加水稀释，用碳酸氢钠溶液中和至中性，用二氯甲烷萃取，浓缩，制备板纯化(石油醚：乙酸乙酯＝1：1)得到白色固体标题化合物(150mg,82％)。LC-MS:m/z[M+H]+＝326.
35：4-(3-(3-(二氟甲基)-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)-4-氟苯基)-7-乙基-7H-咪唑 [4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(80mg,0.22mmol)，3-(二氟甲基)-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(75mg,0.22mmol)为原料，得到白色固体标题化合物(39mg,41％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H)8.38(d,J＝6.85Hz,1H)8.29(s,2H)8.18(d,J＝6.85Hz,1H)7.42(t,J＝9.05Hz,1H)7.30(br.s.,1H)7.08(d,J＝6.85Hz,1H)4.59(q,J＝7.17Hz,2H)4.50(s,3H)1.67-1.70(m,3H).LC-MS:m/z[M+H]+＝440.
实施例36
36-1：2-溴-1-(四氢-2H-吡喃-4-基)乙酮
1-(四氢-2H-吡喃-4-基)乙酮(1000mg 7.8mmol)加到甲醇(5mL)中，降温至0摄氏度，然后加入液溴(0.4ml,7.8mmol),0摄氏度搅拌45分钟，然后室温搅拌45分钟后加入硫酸(2.7ml,30mmol),室温搅拌过夜。反应液加亚硫酸氢钠水溶液淬灭，然后加入乙酸乙酯(300ml)，乙酸乙酯用水洗(200ml x3)，然后干燥后浓缩得到得到标题化合物(600mg,37％)。
36-2：6-溴-7-甲氧基-2-(四氢-2H-吡喃-4-基)咪唑并[1,2-a]吡啶
将5-溴-4-甲氧基吡啶-2-胺(200mg,1mmol),2-溴-1-(四氢-2H-吡喃-4-基)乙酮(600mg,3mmol)加入无水乙醇(10ml)中。80摄氏度搅拌过夜。反应液直接薄层层析(石油醚/乙酸乙酯＝3/1)得到标题化合物(300mg,98％)LC-MS:m/z[M+H]+＝311
36：7-乙基-4-(4-氟-3-(7-甲氧基-2-(四氢-2H-吡喃-4-基)咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑 并[4,5-c]哒嗪
实验操作同实施例2，以6-溴-7-甲氧基-2-(四氢-2H-吡喃-4-基)咪唑并[1,2-a]吡啶(80mg,0.26mmol),7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(50mg,0.13mmol)为原料,得到标题化合物(16mg,25％)。
1H NMR(400MHz,CHLOROFORM-d)1.70(d,J＝7.34Hz,3H)2.07(br.s.,4H)3.08(br.s.,1H)3.59(br.s.,2H)3.89(br.s.,3H)4.07(br.s.,2H)4.58(d,J＝6.85Hz,2H)7.12(br.s.,1H)7.22(br.s.,1H)7.35-7.41(m,1H)8.06(br.s.,1H)8.28(br.s.,3H)9.37(s,1H).LC-MS:m/z[M+H]+＝473
实施例37
37-1：6-溴-2-环丙基-7-甲氧基咪唑并[1,2-a]吡啶
将5-溴-4-甲氧基吡啶-2-胺(2000mg,9.85mmol),2-溴-1-环丙基乙酮(3211mg,19.7mmol)加入无水乙醇(20ml)中。封管，90摄氏度搅拌过夜。反应液直接柱层层析(石油醚/乙酸乙酯＝3/1)得到标题化合物(1900mg,72％)LC-MS:m/z[M+H]+＝267
37：4-(3-(2-环丙基-7-甲氧基咪唑并[1,2-a]吡啶-6-基)-4-氟苯基)-7-乙基-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以6-溴-2-环丙基-7-甲氧基咪唑并[1,2-a]吡啶(150mg,0.56mmol),7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(249mg,0.67mmol)为原料,得到标题化合物(83mg,34％)。
1H NMR(400MHz,DMSO-d6)0.78-0.91(m,4H)1.54(s,3H)1.94-2.00(m,1H)3.79(s,3H) 4.46-4.54(m,2H)6.96(s,1H)7.54(s,2H)8.11-8.16(m,1H)8.47(s,1H)8.50-8.56(m,1H)8.85(s,1H)9.55(s,1H).LC-MS:m/z[M+H]+＝429
实施例38
38-1：3-环丙基-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
实验操作同实施例5中，5-3的合成方法，以2-肼基-4-碘-3-甲氧基吡啶(200mg,0.76mmol)和环丙烷甲醛(53mg,0.76mmol)为原料，得到白色固体标题化合物(160mg,67％)。LC-MS:m/z[M+H]+＝316.
38：4-(3-(3-环丙基-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)-4-氟苯基)-7-乙基-7H-咪唑并 [4],5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(70mg,0.2mmol)，3-环丙基-7-碘-8-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(50mg,0.2mmol)为原料，得到黄色固体标题化合物(43mg,50％)。
1H NMR(400MHz,CHLOROFORM-d)δppm 9.38(s,1H)8.34(d,J＝5.87Hz,1H)8.29(br.s.,2H)7.91(d,J＝6.85Hz,1H)7.39(t,J＝9.05Hz,1H)6.92(d,J＝6.85Hz,1H)4.59(q,J＝6.85Hz,2H)4.38-4.52(m,3H)2.08(br.s.,1H)1.70(t,J＝6.85Hz,3H)1.21(d,J＝7.83Hz,2H)0.88(br.s,2H).LC-MS:m/z[M+H]+＝430.
实施例39
39-1：6-溴-2-(溴甲基)-7-甲氧基咪唑并[1,2-a]吡啶
5-溴-4-甲氧基吡啶-2-胺(300mg,1.48mmol)和二溴丙酮(635mg，2.95mmol)加到无水乙醇(4ml)中，90℃搅拌两个小时。将反应液浓缩，直接上制备板分离纯化(石油醚/乙酸乙酯＝1/1)得标题化合物(210mg，45％)。LC-MS:m/z[M+H]+＝321
39-2：6-溴-7-甲氧基-2-(甲氧基甲基)咪唑并[1,2-a]吡啶
6-溴-2-(溴甲基)-7-甲氧基咪唑并[1,2-a]吡啶(200mg,0.63mmol)和甲醇钠(0.5ml，4mmol)加到甲醇(4ml)中，室温搅拌两个小时。将反应液浓缩，直接上制备板分离纯化(石油醚/乙酸乙酯＝1/1)得标题化合物(35mg，21％)。LC-MS:m/z[M+H]+＝271，273.
39：7-乙基-4-(4-氟-3-(7-甲氧基-2-(甲氧基甲基)咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑 并[4,5-c]哒嗪
实验操作同实施例2，以6-溴-7-甲氧基-2-(甲氧基甲基)咪唑并[1,2-a]吡啶(35mg,0.13mmol)和7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(50mg,0.13mmol)为原料，得标题化合物(21mg，38％)。
1H NMR(400MHz,CDCl3)δ9.37(s,1H),8.31-8.22(m,3H),8.05(s,1H),7.45(s,1H),7.40-7.33(m,1H),7.00(br.s.,1H),4.65-4.54(m,4H),3.87(s,3H),3.48(br.s.,3H),1.69-1.67(m,3H).LC-MS:m/z[M+H]+＝433.
实施例40
40-1：5-溴-6-甲氧基-2-甲基咪唑并[1,2-a]吡啶
将6-溴-5-甲氧基吡啶-2-胺(90mg，0.45mmol)和溴丙酮(120mg，0.88mmol)加到无水乙醇(10mL)中，在90℃反应16小时，浓缩，制备板制备(石油醚：乙酸乙酯＝1：2)得到黄色油状物(35mg,产率32％)。LC-MS:m/z[M+H]+＝241,243.
40：7-乙基-4-(4-氟-3-(6-甲氧基-2-甲基咪唑并[1,2-a]吡啶-5-基)苯基)-7H-咪唑并[4,5-c]哒 嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(54mg,0.15mmol)，5-溴-6-甲氧基-2-甲基咪唑并[1,2-a]吡啶(35mg，0.15mmol)为原料，得到棕色固体(25mg,产率41％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H),8.43(br.s.,2H),8.28(s,2H),7.50(t,J＝9.0Hz,1H),7.36-7.28(m,1H),7.11(br.s.,1H),4.61-4.56(m,2H),3.85(br.s.,3H),2.46-2.32(m,3H),1.71-1.67(m,3H).LC-MS:m/z[M+H]+＝403.
实施例41
41-1：6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-甲醛
5-溴-4-甲氧基吡啶-2-胺(610mg，3.0mmol)，1,1,3-三氯丙酮(720mg，4.5mmol)加到1,2-二甲氧基乙烷(2ml)中，室温搅拌24小时，反应液抽滤，滤得白色固体溶于乙醇(4ml)中，95℃反应24小时，反应液浓缩，加到二氯甲烷(5ml)，三氟醋酸(0.5ml)中，室温搅拌一小时，反应液浓缩，制备薄层色谱(二氯甲烷/甲醇＝10/1)得标题化合物(60mg，7.8％).LC-MS:m/z[M+H]+＝255,257
41-2：6-溴-2-(二氟甲基)-7-甲氧基咪唑并[1,2-a]吡啶
6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-甲醛(60mg，0.235mmol)加到二氯甲烷(2ml)中,(二乙氨基)三氟化硫(0.05ml)加到反应液中，室温搅拌1小时，反应液加入0.5ml淬灭，浓缩，制备薄层色谱(石油醚/乙酸乙酯/二氯甲烷＝3/1/0.1)得标题化合物(17mg，26％)。LC-MS:m/z[M+H]+＝277,279.
41：4-(3-(2-(二氟甲基)-7-甲氧基咪唑并[1,2-a]吡啶-6-基)-4-氟苯基)-7-乙基-7H-咪唑并 [4,5-c]哒嗪
实验操作同实施例19，以6-溴-2-(二氟甲基)-7-甲氧基咪唑并[12-a]吡啶(9mg，0.032mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(12mg，0.032mmol) 为原料，得标题化合物(7mg，50％).
1H NMR(400MHz,CHLOROFORM-d)d＝9.38(s,1H),8.29(br.s.,3H),8.10(s,1H),7.69(s,1H),7.38(s,1H),7.05-6.67(m,2H),4.59(q,J＝7.0Hz,2H),3.88(s,3H),1.70(br.s.,3H).LC-MS:m/z[M+H]+＝439.
实施例42
42-1：5-溴-2-(甲氧羰基)-3-甲基吡啶1-氧化物
将5-溴-3-甲基吡啶甲酸甲酯(1.2g，5.22mmol)和间氯过氧苯甲酸(3g，17.44mmol)加到二氯甲烷(100mL)中，在室温搅拌16小时，浓缩，柱层析(石油醚：乙酸乙酯＝5：1)得到标题化合物白色固体(1.06g,产率83％)。LC-MS:m/z[M+H]+＝246,248.
42-2：5-溴-6-氯-3-甲基吡啶甲酸甲酯
将5-溴-2-(甲氧羰基)-3-甲基吡啶1-氧化物(1.05g，4.30mmol)和三乙胺(5g，49.5mmol)加到二氯甲烷(100mL)中，冰浴下滴加草酰氯(2.5g，20.5mmol)在室温搅拌1小时，过滤，母液浓缩，柱层析(石油醚：乙酸乙酯＝5：1)得到标题化合物浅黄色固体(0.52g,产率44％)。LC-MS:m/z[M+H]+＝264,266.
42-3：5-溴-3-(溴甲基)-6-氯吡啶甲酸甲酯
将5-溴-6-氯-3-甲基吡啶甲酸甲酯(0.52g，1.90mmol)、NBS(0.48g，2.7mmol)和AIBN(30mg)加到四氯化碳(10mL)中，65℃下，搅拌16小时，浓缩，制备板制备(石油醚：乙酸乙酯＝5：1)得到含标题化合物和原料的混合物(0.5g,1:1粗品)。LC-MS:m/z[M+H]+＝342,344,346.
42-4：3-溴-2-氯-6-甲基-5,6-二氢-7H-吡咯并[3,4-b]吡啶-7-酮
将5-溴-3-(溴甲基)-6-氯吡啶甲酸甲酯(0.5g，粗品)、甲胺盐酸盐(200mg，3mmol)和三乙胺(0.5g，4.95mmol)加到乙腈(10mL)中，在室温搅拌2小时，加入氯化铵饱和溶液100mL，乙酸乙酯萃取(3×100mL)，母液浓缩，制备板制备得到标题化合物白色固体(200mg)。LC-MS:m/z[M+H]+＝261,263.
42-5：3-溴-2-甲氧基-6-甲基-5,6-二氢-7H-吡咯并[3,4-b]吡啶-7-酮
将3-溴-2-氯-6-甲基-5,6-二氢-7H-吡咯并[3,4-b]吡啶-7-酮(120mg，0.46mmol)、磷酸钠(250mg，1.52mmol)加到甲醇(5mL)中，60℃下，搅拌16小时，过滤，母液浓缩，制备板制备得到标题化合物白色固体(17mg，14％)。LC-MS:m/z[M+H]+＝257,259.
42：3-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-2-甲氧基-6-甲基-5,6-二氢-7H- 吡咯并[3,4-b]吡啶-7-酮
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(25mg,0.07mmol)，3-溴-2-甲氧基-6-甲基-5,6-二氢-7H-吡咯并[3,4-b]吡啶-7-酮 (15mg，0.06mmol)为原料，得到棕色固体(12mg,产率48％)。
1H NMR(400MHz,CHLOROFORM-d)9.37(s,1H),8.28(s,3H),7.78(s,1H),7.37(t,J＝9.3Hz,1H),4.67-4.52(m,2H),4.38(s,2H),4.10(s,3H),3.28(s,3H),1.71-1.68(m,3H).LC-MS:m/z[M+H]+＝419.
实施例43
43-1：6-溴-7-甲氧基-2-(三氟甲基)咪唑并[1,2-a]吡啶
5-溴-4-甲氧基吡啶-2-胺(202mg，1.0mmol)加到乙醇(1.5ml)和1,4-二氧六环(0.5ml)混合溶剂中，3-氯-1,1,1-三氟丙酮(161mg，1.1mmol)缓慢加到反应液，室温搅拌6小时，三乙胺(202mg，2.0mmol)加到反应液中，95℃反应过夜，反应液抽滤，浓缩，制备薄层色谱(石油醚/乙酸乙酯＝20/1)得标题化合物(50mg，16.9％).LC-MS:m/z[M+H]+＝295,297.
43：7-乙基-4-(4-氟-3-(7-甲氧基-2-(三氟甲基)咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑并 [4,5-c]哒嗪
实验操作同实施例19，以6-溴-7-甲氧基-2-(三氟甲基)咪唑并[1,2-a]吡啶(50mg，0.170mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(58mg，0.155mmol)为原料，得标题化合物(32mg，45％).
1H NMR(400MHz,CHLOROFORM-d)9.36(br.s.,1H),8.27(br.s.,3H),8.09(s,1H),7.76(s,1H),7.36(t,J＝8.6Hz,1H),7.01(s,1H),4.57(d,J＝7.3Hz,2H),3.86(s,3H),1.68(br.s.,3H).LC-MS:m/z[M+H]+＝457.
实施例44
44-1：4-溴-2-肼基-5-甲氧基吡啶
将4-溴-2-氟-5-甲氧基吡啶(560mg,2.72mmol)和水合肼(1360mg,27.20mmol)加到乙醇(10mL)中，在80℃下搅拌16小时。反应液浓缩，制备薄层色谱(乙酸乙酯)分离得黄色油状液体标题化合物(110mg,20％)。LC-MS:m/z[M+H]+＝218,220.
44-2：7-溴-6-甲氧基-[1,2,4]三唑并[4,3-a]吡啶
将4-溴-2-肼基-5-甲氧基吡啶(110mg,0.50mmol)加到甲酸(5mL)中，在110℃下搅拌16小时后。反应液过滤后浓缩，制备薄层色谱(二氯甲烷/甲醇＝20/1)分离得白色固体标题化合物(30mg,26％)。LC-MS:m/z[M+H]+＝228,230.
44：7-乙基-4-(4-氟-3-(6-甲氧基-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-溴-6-甲氧基-[1,2,4]三唑并[4,3-a]吡啶(30mg,0.13mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(48mg,0.13mmol)为原料，得黄色固体标题化合物(3mg,6％)。
1H NMR(400MHz,CHLOROFORM-d)9.37(s,1H),8.83(s,1H),8.34(d,J＝6.4Hz,1H),8.28(s,2H),7.82(s,1H),7.71(s,1H),7.37(t,J＝9.0Hz,1H),4.58(q,J＝7.3Hz,2H),3.86(s,3H),1.69(t,J＝7.3Hz,3H).LC-MS:m/z[M+H]+＝390.
实施例45
45-1：(6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-基)甲醇
将6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-羧酸甲酯(800mg,2.81mmol)加到二氯甲烷(4mL)中，再加入二异丁基氢化铝的正己烷溶液(1M,4mL)，在室温下搅拌16小时后。反应液过滤后浓缩，制备薄层色谱(二氯甲烷/甲醇＝10/1)分离得黄色固体标题化合物(190mg,26％)。LC-MS:m/z[M+H]+＝257,259.
45：(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑并[1,2-a]吡啶-2-基)甲醇
实验操作同实施例2，以(6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-基)甲醇(50mg,0.19mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(70mg,0.19mmol)为原料，得棕色固体标题化合物(15mg,18％)。
1H NMR(400MHz,CHLOROFORM-d)9.23(s,1H),8.32(s,1H),8.20-8.10(m,3H),7.45(s,1H),7.30(t,J＝9.3Hz,1H),7.20-6.98(m,1H),4.67(s,2H),4.50(q,J＝7.2Hz,2H),3.80(s,3H),1.60(t,J＝7.3Hz,3H).LC-MS:m/z[M+H]+＝419.
实施例46
46-1：6-溴-7-甲氧基咪唑并[1,2-a]吡啶-3-甲醛
将5-溴-4-甲氧基吡啶-2-胺(300mg,1.48mmol)和2-溴丙二醛(335mg,2.22mmol)加到乙醇(10mL)中，在80℃下搅拌0.5小时后，再加入碳酸氢钠(249mg,2.96mmol)在80℃下搅拌16小时。反应液过滤后浓缩，制备薄层色谱(乙酸乙酯)分离得黄色固体标题化合物(120mg,25％)。LC-MS:m/z[M+H]+＝255,257.
46-2：6-溴-7-甲氧基咪唑并[1,2-a]吡啶-3-甲醇
将6-溴-7-甲氧基咪唑并[1,2-a]吡啶-3-甲醛(120mg,0.47mmol)加到二氯甲烷(4mL)中，再加 入二异丁基氢化铝的正己烷溶液(1M,4mL)，在室温下搅拌16小时后。反应液过滤后浓缩，制备薄层色谱(二氯甲烷/甲醇＝10/1)分离得白色固体标题化合物(40mg,33％)。LC-MS:m/z[M+H]+＝257,259.
46：(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑并[1,2-a]吡啶-3-基)甲 醇
实验操作同实施例2，以6-溴-7-甲氧基咪唑并[1,2-a]吡啶-3-甲醇(40mg,0.16mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(59mg,0.16mmol)为原料，得棕色固体标题化合物(6mg,8％)。
1H NMR(400MHz,DMSO-d6)9.57-9.54(m,1H),8.87-8.84(m,1H),8.59-8.53(m,1H),8.53-8.47(m,2H),7.58-7.53(m,1H),7.52-7.49(m,1H),7.26-7.15(m,1H),5.33-5.26(m,1H),4.79-4.75(m,2H),4.53-4.47(m,2H),3.86(s,3H),1.54(s,3H).LC-MS:m/z[M+H]+＝419.
实施例47
47-1：3-溴-1,8-萘啶-2(1H)-酮
2-氧-1,2-二氢-1,8-萘啶-3-羧酸(490mg,2.58mmol)加到N，N-二甲基甲酰胺(10mL)中，加入液溴(2g,12.9mmol)，吡啶(5ml)，100摄氏度搅拌1小时。反应液中滴加5ml饱和亚硫酸氢钠水溶液，然后加乙酸乙酯萃取(3 x 200ml)，乙酸乙酯用水洗三次后用饱和食盐水洗一次，然后用无水硫酸钠干燥，浓缩后粗产品用薄层析纯化(石油醚/乙酸乙酯＝4/1)得到标题化合物(140mg,21％)。LC-MS:m/z[M+H]+＝225
47-2：3-溴-2-甲氧基-1,8-萘啶
3-溴-1,8-萘啶-2(1H)-酮(140mg,0.63mmol)加到二氯甲烷/甲醇(5mL/2ml)中，然后加三甲基硅烷化重氮甲烷(2ml)，室温搅拌过夜。反应液直接浓缩，然后用薄层析纯化(石油醚/乙酸乙酯＝4/1)得到标题化合物(40mg,27％)。LC-MS:m/z[M+H]+＝239
47：3-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-2-甲氧基-1,8-萘啶
实验操作同实施例19，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(74mg,0.2mmol)，3-溴-2-甲氧基-1,8-萘啶(40mg,0.17mmol)为原料,得到标题化合物(38mg,48％).
1H NMR(400MHz,CHLOROFORM-d)9.39(s,1H)8.66(d,J＝3.42Hz,1H)8.43(d,J＝4.89Hz,1H)8.29(br.s.,1H)8.27(s,1H)7.95(d,J＝6.36Hz,1H)7.90(s,1H)7.39(t,J＝9.05Hz,1H)7.23(d,J＝7.83Hz,1H)4.58(q,J＝7.34Hz,2H)3.94(s,3H)1.69(t,J＝7.34Hz,3H).LC-MS:m/z[M+H]+＝401.
实施例48
48-1：6-溴-7-甲氧基咪唑并[1,2-a]嘧啶
5-溴-4-甲氧基嘧啶-2-胺(203mg，1.0mmol)，氯乙醛(94mg，1.2mmol)加到乙醇(2ml)中，室温搅拌过夜。碳酸氢钠(168mg，2.0mmol)加到反应液，60℃反应2小时.反应液抽滤，浓缩，制备薄层色谱(二氯甲烷/甲醇＝20/1)得标题化合物(57mg，25％).LC-MS:m/z[M+H]+＝228,230.
48：7-乙基-4-(4-氟-3-(7-甲氧基咪唑并[1,2-a]嘧啶-6-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例19，以6-溴-7-甲氧基咪唑并[1,2-a]嘧啶(57mg，0.251mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(92mg，0.251mmol)为原料，得标题化合物(50mg，52％).
1H NMR(400MHz,CHLOROFORM-d)9.37(s,1H),8.31(d,J＝16.6Hz,4H),7.57(br.s.,1H),7.44-7.34(m,2H),4.59(q,J＝7.3Hz,2H),4.08(s,3H),1.72-1.68(m,3H).LC-MS:m/z[M+H]+＝390.
实施例49
49-1：5-溴-6-甲氧基咪唑并[1,2-a]吡啶
将6-溴-5-甲氧基吡啶-2-胺(200mg，0.99mmol)和氯乙醛(40％，290mg，1.49mmol)加到无水甲醇(10mL)中，室温下搅拌1小时，再加入碳酸氢钠(200mg，2.38mmol)，在60℃反应16小时，浓缩，制备板制备(石油醚：乙酸乙酯＝3：1)得到浅黄色固体(130mg,产率58％)。LC-MS:m/z[M+H]+＝227,229.
49：7-乙基-4-(4-氟-3-(6-甲氧基咪唑并[1,2-a]吡啶-5-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(36mg,0.1mmol)，5-溴-6-甲氧基咪唑并[1,2-a]吡啶(23mg，0.1mmol)为原料，得到棕色固体(7mg,产率18％)。
1H NMR(400MHz,CHLOROFORM-d)9.38(s,1H),8.47-8.39(m,2H),8.28(s,1H),7.84-7.59(m,1H),7.50(t,J＝8.8Hz,1H),7.41-7.26(m,2H),7.25-6.97(m,1H),4.58(q,J＝7.3Hz,2H),3.85(s,3H),1.70-1.65(m,3H).LC-MS:m/z[M+H]+＝389.
实施例50
50：7-乙基-4-(4-氟-3-(7-甲氧基-2-甲基咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以6-溴-7-甲氧基-2-甲基咪唑并[1,2-a]吡啶(30mg,0.12mmol)，7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(44mg,0.12mmol)为原料，得白色固体标题化合物(13mg,27％)。
1H NMR(400MHz,CHLOROFORM-d)9.35(s,1H),8.32-8.16(m,3H),7.98(s,1H),7.34(t,J＝8.8Hz,1H),7.20(s,1H),6.90(br.s.,1H),4.57(q,J＝7.3Hz,2H),3.93-3.74(m,3H),2.47-2.33(m,3H),1.68(t,J＝7.1Hz,3H).LC-MS:m/z[M+H]+＝403.
实施例51
51-1：6-溴-5-甲氧基咪唑并[1,2-a]吡啶
将5-溴-6-甲氧基吡啶-2-胺(200mg，0.99mmol)和氯乙醛(40％，290mg，1.49mmol)加到无水甲醇(10mL)中，室温下搅拌1小时，再加入碳酸氢钠(200mg，2.38mmol)，在70℃反应16小时，浓缩，制备板制备(石油醚：乙酸乙酯＝3：1)得到浅黄色固体(70mg,产率31％)。LC-MS:m/z[M+H]+＝227,229.
51：7-乙基-4-(4-氟-3-(5-甲氧基咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(36mg,0.1mmol)，6-溴-5-甲氧基咪唑并[1,2-a]吡啶(23mg，0.1mmol)为原料，得到棕色固体(4mg,产率10％)。
1H NMR(400MHz,CHLOROFORM-d)9.39(s,1H),8.41(d,J＝4.9Hz,1H),8.29(br.s.,2H),7.76(br.s.,2H),7.49-7.27(m,3H),4.68-4.50(m,2H),3.84(s,3H),1.69(br.s.,3H).LC-MS:m/z[M+H]+＝389.
实施例52
52：7-乙基-4-(4-氟-3-(7-甲氧基咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑并[4,5-c]哒嗪
实验操作同实施例2，以7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(50mg,0.14mmol)，6-溴-7-甲氧基咪唑并[1,2-a]吡啶(30mg,0.14mmol)为原料，得到黄色固体标题化合物(8mg,15％)。
1H NMR(400MHz,DMSO-d6)δ9.57(s,1H)8.87(s,1H)8.67(s,1H)8.48-8.61(m,2H)7.82(br.s.,1H)7.46-7.65(m,2H)7.17(br.s.,1H)4.51(q,J＝6.85Hz,2H)3.84(s,3H)1.55(t,J＝7.09Hz,3H).LC-MS:m/z[M+H]+＝389.
实施例53
53-1：N-(4-(甲氧基甲基)吡啶-2-基)-1,1-二苯基甲胺
将2-氯-4-(甲氧基甲基)吡啶(2.87g,18.28mmol)，二苯甲胺(3.3g,18.28mmol)，R-(+)-1.1-联联萘-2,2-二苯膦(1.10g,1.82mmol)，碳酸铯(8.30g,25.59mmol)加入到1，4-二氧六环(50ml)溶液中，换3次氩气，加热至80℃反应15小时。冷却至室温，将反应液倒入冰水(40ml)中，乙酸乙酯(50mL×3)萃取，用饱和氯化钠水溶液(50mL×3)水洗，有机相干燥，浓缩经过prep-HPLC制备纯化得到(3.4g，61.8％)黄色油状液体。LC-MS:m/z[M+1]+:303
53-2：4-(甲氧基甲基)吡啶-2-胺
将N-(4-(甲氧基甲基)吡啶-2-基)-1,1-二苯基甲胺(3.3g,11mmol)溶于四氢呋喃(50ml)溶液中，再加入盐酸的1，4-二氧六环(17ml)，室温下反应2h,反应完全后，往反应液中加入饱和碳酸氢钠(30ml)中调节PH＝7左右，乙酸乙酯(50mLx3)萃取，用饱和氯化钠水溶液(50mLx3)水洗，有机相干燥，浓缩经过prep-HPLC制备纯化得到(800mg，53.3％)白色固体。LC-MS:m/z[M+1]+:139
53-3：5-溴-4-(甲氧基甲基)吡啶-2-胺
将5-溴-4-(甲氧基甲基)吡啶-2-胺(260mg,1.88mmol)，乙酸铵(14.5mg,0.188mmol)，加入到乙腈(25ml)溶液中，在温度0℃下分批加入NBS(167.7mg,0.94mmol)，撤走冰水浴，室温反应40分钟。加入(20ml)二氯甲烷溶解，饱和碳酸氢钠(40ml)调节PH＝7左右，二氯甲烷(50mlx4)萃取,有机相干燥，浓缩经过prep-HPLC制备纯化得到(220mg，40％)白色固体。LC-MS:m/z[M+1]+:219
53-4：6-溴-2-(1-氟环丙基)-7-(甲氧基甲基)咪唑并[1,2-a]吡啶
将5-溴-4-(甲氧基甲基)吡啶-2-胺(524mg,2.5mmol)，2-氯-1-(1-氟环丙基)乙-1-酮(670mg,5mmol)，碳酸铯(1.6g,5mmol)溶于乙醇(10.0mL)的溶液中，在微波反应器中加热到100℃反应1小时,将反应液经过制备HPLC纯化得到(100mg，26％)白色固体。LC-MS:m/z[M+1]+＝300
53：7-乙基-4-(4-氟-3-(2-(1-氟环丙基)-7-(甲氧基甲基)咪唑并[1,2-a]吡啶-6-基)苯基)-7H-咪唑并[4,5-c]哒嗪
将6-溴-2-(1-氟环丙基)-7-(甲氧基甲基)咪唑并[1,2-a]吡啶(50.0mg,0.17mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(105mg,0.36mmol)为原料，得到标题化合物(20mg，26％)类 白色固体。
1H NMR(400MHz,CHLOROFORM-d)d＝9.39(s,1H),8.30(d,J＝10.8Hz,3H),8.13(br.s.,1H),7.81(br.s.,1H),7.70(s,1H),7.40(t,J＝8.6Hz,1H),4.59(q,J＝7.0Hz,2H),4.38(s,2H),3.31(s,3H),1.70(t,J＝7.1Hz,3H),1.54(d,J＝19.6Hz,2H),1.26(d,J＝14.5Hz,2H).LC-MS:m/z[M+1]+＝461
实施例54
54-1：(2,4-二氯吡啶-3-基)甲醇
在氮气保护下将2，4-二氯烟醛(5g，28.6mmol)溶解在50ml乙醇中，将溶液降温至0℃，在此温度下，缓慢加入硼氢化钠(1.4g，37.2mmol)，恢复至室温，反应16小时，加入冰的氯化铵水溶液中，二氯甲烷萃取(100ml x3),有机相浓缩，得(2,4-二氯吡啶-3-基)甲醇(4.8g,94.1％)黄色固体。LC-MS:m/z[M+H]+＝178。
54-2：2,4-二氯-3-(甲氧基甲基)吡啶
在氮气保护下，将(2,4-二氯吡啶-3-基)甲醇(4.8g，27mmol)溶解在50ml N,N-二甲基甲酰胺中，将溶液降温至0℃，在此温度下，缓慢加入NaH(2.2g，54mmol)。在冰浴下反应半小时，再加入碘甲烷(4.6g，32.4mmol)，室温16小时，反应结束后将反应液加入100ml冰水中，乙酸乙酯萃取(100ml x3),水(50mLx2)反洗，有机相旋干，得到2,4-二氯-3-(甲氧基甲基)吡啶(2.3g，44.5％)淡黄色液体。1H NMR(400MHz,CDCl3)8.16(d,J＝5.1Hz,1H),7.23(d,J＝5.2Hz,1H),4.64(s,2H),3.38(s,3H).LC-MS:m/z[M+H]+＝192
54-3：N-(4-氯-3-(甲氧基甲基)吡啶-2-基)-1,1-二苯基甲基苯胺
在氮气保护下，将2,4-二氯-3-(甲氧基甲基)吡啶(1g,5.8mmol)，二苯甲胺(1.1g，5.8mmol)溶解在15mL，再加入1,1'-联萘-2,2'-双二苯膦(361mg，0.58mmol)，碳酸铯(2.83g，8.7mmol)，最后加入醋酸钯(130mg.0.58mmol)在90℃反应16小时。反应结束后，将反应液加入60ml水中，用乙酸乙酯萃取(50mLx3),有机相旋干，柱层析(石油醚/乙酸乙酯＝10/1),旋干有机相得到N-(4-氯-3-(甲氧 基甲基)吡啶-2-基)-1，1-二苯基甲基苯胺(700mg，40％)黄色油状液体。LC-MS:m/z[M+H]+＝337
54-4：4-氯-3-(甲氧基甲基)吡啶-2-胺
在氮气保护下，将N-(4-氯-3-(甲氧基甲基)吡啶-2-基)-1,1-二苯基甲基苯胺(0.7g，2.1mmol)加入10ml四氢呋喃中，再加入8ml盐酸二氧六环溶液。室温下16小时，反应结束后，将反应液加入20ml碳酸氢钠溶液中，乙酸乙酯萃取(30ml x3)，旋干有机相，柱层析(石油醚/乙酸乙酯＝2/1)，得到4-氯-3-(甲氧基甲基)吡啶-2-胺(220mg,20％)白色固体。
1HNMR(400MHz,CDCl3)7.85(d,J＝5.7Hz,1H),6.71(d,J＝5.8Hz,1H),5.72(s,2H),4.69(s,2H),3.37(s,3H).LC-MS:m/z[M+H]+＝173
54-5：7-氯-8-(甲氧基甲基)-3-甲基咪唑并[1,2-a]吡啶
在氮气保护下，将4-氯-3-(甲氧基甲基)吡啶-2-胺(50mg，0.29mmol)，1,1-二甲氧基-2-溴丙烷(265mg，1.45mmol)溶解在2mL乙醇中，再加入0.2ml盐酸甲醇溶液，微波120℃反应2小时。直接旋干用于下一步，得到7-氯-8-(甲氧基甲基)-3-甲基咪唑并[1,2-a]吡啶(70mg,100％)白色固体。LC-MS:m/z[M+H]+＝211
54：7-乙基-4-(4-氟-3-(8-(甲氧基甲基)-3-甲基咪唑[1,2-a]吡啶-7-基)苯基)-7H-咪唑[4,5-c]哒嗪
实验操作同实施例2，以7-氯-8-(甲氧基甲基)-3-甲基咪唑并[1,2-a]吡啶(70mg,0.33mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(95mg,0.33mmol)为原料，得标题化合物(16.1mg,11.6％)白色固体。
1HNMR(400MHz,DMSO-d6)9.54(s,1H),8.87(s,1H),8.61–8.53(m,2H),8.35(d,J＝7.1Hz,1H),7.59(t,J＝9.3Hz,1H),7.46(d,J＝0.7Hz,1H),7.00(d,J＝7.0Hz,1H),4.72(s,2H),4.52(q,J＝7.3Hz,2H),3.11(d,J＝3.8Hz,3H),2.53(s,3H),1.64–1.47(m,3H).LC-MS:m/z[M+H]+＝417
实施例55和实施例56
55-1：1-(6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-基)乙烷-1-酮
将5-溴-4-甲氧基吡啶-2-胺(500mg,2.46mmol)，1-溴丁烷-2,3-二酮(406mg,2.46mmol)加到乙醇(10mL)中，90摄氏度搅拌过夜。反应液直接薄层层析(石油醚/乙酸乙酯＝1/1)得到黄色固体标题化合 物(150mg,22.66％)。LC-MS:m/z[M+H]+＝269
55-2：1-(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑并[1,2-a]吡啶-2-基) 乙烷-1-酮
实验操作同实施例2，以1-(6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-基)乙烷-1-1(150mg,0.56mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(161mg,0.56mmol)为原料，得到黄色固体标题化合物(100mg,41.29％)。LC-MS:m/z[M+H]+＝431
55：(Z)-1-(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑[1,2-a]吡啶-2-基) 乙烷-1-酮氧-甲基肟
56：(E)-1-(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑并[1,2-a]吡啶-2- 基)乙烷-1-酮氧-甲基肟
将1-(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑并[1,2-a]吡啶-2-基)乙烷-1-酮(100mg,0.23mmol),甲氧基胺盐酸盐(58mg,0.69mmol)加入乙醇(10ml)中。室温搅拌1小时。反应液直接薄层析纯化(二氯甲烷/甲醇＝20/1)得到黄色固体实施例55(6mg,5％)和实施例56(15mg,13％)。
实施例55：1H NMR(400MHz,DMSO-d6)ppm 1.50-1.57(m,3H)2.26-2.32(m,3H)3.80-3.85(m,3H)3.89-3.97(m,3H)4.47-4.54(m,2H)7.12-7.17(m,1H)7.52-7.58(m,1H)8.44-8.59(m,3H)8.65-8.71(m,1H)8.83-8.89(m,1H)9.52-9.60(m,1H).LC-MS:m/z[M+H]+＝460
实施例56：1H NMR(400MHz,DMSO-d6)ppm 1.54(t,J＝7.34Hz,3H)2.21-2.29(m,3H)3.83(s,3H)3.89(s,3H)4.49-4.53(m,2H)7.11(s,1H)7.54(t,J＝8.80Hz,1H)8.02(s,1H)8.48-8.60(m,3H)8.86(s,1H)9.56(s,1H).LC-MS:m/z[M+H]+＝460
实施例57
57-1：2-(6-溴-7-甲氧基[1,2-a]吡啶-2-基)丙-2-醇
将甲基氯化镁(620mg,8.35mmol)和6-溴-7-甲氧基咪唑并[1,2-a]吡啶-2-羧酸甲酯(500mg,1.67mmol)加到四氢呋喃(20mL)中，在室温下搅拌1小时。将反应液用盐酸水溶液(1N)调至酸性，浓缩，再用饱和碳酸氢钠水溶液调至碱性，用二氯甲烷萃取(50mL×3)，有机相用饱和食盐水(50mL)洗涤后，再用硫酸钠干燥，浓缩后经制备薄层色谱(二氯甲烷/甲醇＝20/1)分离得标题化合物(130mg,27％)。LC-MS:m/z[M+H]+＝285,287.
57：2-(6-(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)-7-甲氧基咪唑并[1,2-a]吡啶-2-基)丙 -2-醇
实验操作同实施例2，以2-(6-溴-7-甲氧基[1,2-a]吡啶-2-基)丙-2-醇(120mg,0.42mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(120mg,0.42mmol)为原料，得标题化合物(25mg,13％)。
1H NMR(400MHz,CHLOROFORM-d)9.37(s,1H),8.31-8.23(m,3H),8.03(s,1H),7.40-7.33(m,2H),6.97(s,1H),4.59(q,J＝7.3Hz,2H),3.86(s,3H),1.71-1.67(m,9H).LC-MS:m/z[M+H]+＝447.
实施例58
58-1：4-溴-3-(((叔丁基二甲基甲硅烷基)氧基)甲基)-2-氟吡啶
(4-溴-2-氟吡啶-3-基)甲醇(2.0g,9.71mmol)，叔丁基氯二甲基硅烷(2.93g,19.42mmol)，咪唑(1.32g,19.42mmol)加到二氯甲烷(15ml)中，室温搅拌30分钟.反应液拌样，柱层析(石油醚/乙酸乙酯＝20/1)得标题化合物无色油状液体(3.8g,粗品)。LC-MS:m/z[M+H]+＝320,322.
58-2：4-溴-3-(((叔丁基二甲基甲硅烷基)氧基)甲基)-2-肼基吡啶
4-溴-3-(((叔丁基二甲基甲硅烷基)氧基)甲基)-2-氟吡啶(3.8g,11.86mmol)，水合肼(5.94g,118.6mmol)加到1,4-二氧六环(20ml)中，80℃反应2小时，反应液拌样，柱层析(石油醚/乙酸乙酯/氨水＝20/1/0.05)得标题化合物灰色固体(2.1g,53％)。LC-MS:m/z[M+H]+＝332,334.
58-3：7-溴-8-(((叔丁基二甲基甲硅烷基)氧基)甲基)-3-甲基-[1,2,4]三唑并[4,3-a]吡啶
4-溴-3-(((叔丁基二甲基甲硅烷基)氧基)甲基)-2-肼基吡啶(1.03g,3.01mmol)，5M乙醛四氢呋喃溶液(1.6ml)加到1,4-二氧六环(6ml)中，80℃反应1小时，溴化铜(280mg,1.24mmol)，单过硫酸氢钾(2.29g,3.72mmol)加到反应液，室温反应4小时。反应液抽滤，拌样，柱层析(二氯甲烷/甲醇＝30/1)得标题化合物白色固体(262mg,23.54％)。LC-MS:m/z[M+H]+＝356,358.
58-4：(7-溴-3-甲基-[1,2,4]三唑并[4,3-a]吡啶-8-基)甲醇
7-溴-8-(((叔丁基二甲基甲硅烷基)氧基)甲基)-3-甲基-[1,2,4]三唑并[4,3-a]吡啶(260mg,0.61mmol)加到二氯甲烷(10ml)中，4M盐酸二氧六环溶液(2ml)加入，室温搅拌20分钟.反应液浓缩，拌样，柱层析(二氯甲烷/甲醇＝20/1)得标题化合物棕色固体(160mg,96％).LC-MS:m/z[M+H]+＝242,244.
58-5：7-溴-8-(甲氧基甲基)-3-甲基-[1,2,4]三唑并[4,3-a]吡啶
(7-溴-3-甲基-[1,2,4]三唑并[4,3-a]吡啶-8-基)甲醇(160mg,0.66mmol)加到N,N-二甲基甲酰胺(2ml)中，氢化钠(52mg,1.32mmol)加到反应液，室温搅拌15分钟，碘甲烷(190mg,1.31mmol)加到反应液，室温反应30分钟。反应液制备薄层色谱(二氯甲烷/甲醇＝20/1)得标题化合物(143mg,85％)。LC-MS:m/z[M+H]+＝256,258.
58：7-乙基-4-(4-氟-3-(8-(甲氧基甲基)-3-甲基-[1,2,4]三唑并[4,3-a]吡啶-7-基)苯基)-7H-咪唑并 [4,5-c]哒嗪
实验操作同实施例1，以7-溴-3-甲基-8-(甲氧基甲基)-[1,2,4]三唑并[4,3-a]吡啶(143mg,0.56mmol)，(5-(7-乙基-7H-咪唑并[4,5-c]哒嗪-4-基)-2-氟苯基)硼酸(159mg,0.56mmol)为原料，得标题化合物棕色固体(82mg,35％).
1H NMR(400MHz,CHLOROFORM-d)9.40(s,1H),8.43(d,J＝5.9Hz,2H),8.28(s,1H),7.89(d,J＝7.3Hz,1H),7.42(t,J＝9.3Hz,1H),6.95(d,J＝6.8Hz,1H),4.84(s,2H),4.59(q,J＝7.3Hz,2 H),3.41(s,3H),2.81(s,3H),1.70(t,J＝7.3Hz,3H).LC-MS:m/z[M+H]+＝418.
实施例59
59-1：3-甲氧基苯并[b]噻吩
3-溴苯并[b]噻吩(500mg,2.35mmol)，甲醇钠(1267mg,23.5mmol)，氧化铜(93mg,1.18mmol)，碘化亚铜(22mg,0.12mmol)，碘化钾(19mg,0.12mmol)加到N,N-二甲基甲酰胺(5ml)，甲醇(5ml)中，密封管中120℃反应过夜。反应液加入水(30ml)，乙酸乙酯萃取，有机相合并，饱和氯化钠水溶液洗，无水硫酸钠干燥，浓缩，柱层析(纯石油醚)得标题化合物无色油状液体(308mg,80％)。LC-MS:m/z[M+H]+＝165.
59-2：2-溴-3-甲氧基苯并[b]噻吩
3-甲氧基苯并[b]噻吩(308mg,1.88mmol)，N-溴代丁二酰亚胺(368mg,2.07mmol)加到二氯甲烷(5ml)中，25℃搅拌30分钟。反应液抽滤，浓缩，制备薄层色谱(纯石油醚)得标题化合物粉色油状液体(311mg,68％)。1H NMR(400MHz,CHLOROFORM-d)d＝7.72(d,J＝7.3Hz,1H),7.66(d,J＝7.8Hz,1H),7.39-7.31(m,2H),4.05(s,3H).
59：7-乙基-4-(4-氟-3-(3-甲氧基苯并[b]噻吩-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪
7-乙基-4-(4-氟-3-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)苯基)-7H-咪唑并[4,5-c]哒嗪(59mg,0.16mmol)，2-溴-3-甲氧基苯并[b]噻吩(97mg,0.4mmol)，碳酸铯(104mg,0.32mmol)，四(三苯基膦)钯(9mg,0.008mmol)加到1,4-二氧六环(2ml)和水(0.2ml)中，氩气保护下，100℃反应4小时。反应液抽滤，浓缩，制备薄层色谱(石油醚/二氯甲烷/甲醇＝20/30/1)得标题化合物淡黄色固体(6mg,9％)。
1H NMR(400MHz,CHLOROFORM-d)9.43(br.s.,1H),8.68(d,J＝5.4Hz,1H),8.31(br.s.,2H),7.90-7.76(m,2H),7.41(br.s.,3H),4.60(d,J＝7.3Hz,2H),3.93(br.s.,3H),1.70(t,J＝6.4Hz,3H).LC-MS:m/z[M+H]+＝405.
细胞系构建和传代培养
α亚基、β亚基和γ亚基对形成一个完整的功能型GABAA受体是必不可少的。在该实施例中，本申请通过脂质体转染法(Felgner,P.L.,et al.Proceedings of the National Academy of Sciences,1987,84:7413-7417)构建并分别筛选出稳定表达人源α1-GABAA受体(α1-GABAAR)和α2-GABAA受体(α2-GABAAR)的HEK293细胞。α1-GABAAR-HEK293细胞模型同时表达α1亚基(蛋白序列见GenBank登录号：NM_000806.5)、β3亚基(蛋白序列见GenBank登录号：NM_000814.5)和γ2亚基(蛋白序列见GenBank登录号：NM_000816.3)。α2-GABAAR-HEK293细胞模型同时表达α2亚基(蛋白序列见GenBank登录号：NM_000807.4)、β3亚基(蛋白序列见GenBank登录号：NM_000814.5)和γ2亚基(蛋白序列见GenBank登录号：NM_000816.3)。
将以上细胞系进行传代培养。扩增α2-GABAAR-HEK293细胞用于化合物对GABAA受体的苯二氮卓位点(BZD)亲和力测试。α1-GABAAR-HEK293细胞和α2-GABAAR-HEK293细胞在传代过程中部分悬浮细胞铺在用Poly-D-Lysine(多聚赖氨酸)预处理的玻片上用于电生理测试(方法参照专利CN 107344936A第0586段)。
本申请化合物对α2-GABAA受体的亲和活性
通过竞争3H-flunitrazepam(同位素3H标记的氟硝西泮)与稳定表达人源α2-GABAAR-HEK293细胞的膜蛋白BZD位点结合来测定化合物对α2-GABAA受体的亲和力。
膜制备：细胞悬浮于50mM Tris-HCl缓冲液(pH＝7.4)中，在冰上经匀浆机匀浆20秒10次，4℃,1000g离心10min。取上清，重复以上步骤。将上清在4℃(33800g；Thermo(赛默飞)， rotor(转子)：A27-8x50)离心60min，取沉淀重悬于Tris缓冲液(50mM Tris-HCl,10mM MgCl2,0.5mM EDTA，10％glycerol)中。测定蛋白质含量(基于铜离子还原法的BCA(二辛可宁酸)蛋白定量测试法,BCA试剂盒购自Pierce(安诺伦生物))，制备1mL等分物，-80℃保存。
放射性配体竞争结合试验：本试验在在200μL体系中(96孔板)进行，其中有装有100μL细胞膜。3H-flunitrazepam的浓度为1nM，待测化合物的浓度在1x 10-5-10-6M范围内。以flumazenil(氟马泽尼)为对照。低信号对照孔(Low control，LC)加入1μL 2mM flumazenil(终浓度10μM)，高信号对照孔(High control，HC)加入1μL二甲基亚砜。最终靶点膜蛋白的浓度为5μg/孔。所有待测化合物样品储存液均为10mM二甲基亚砜溶液。样品工作浓度为将所有样品用二甲基亚砜稀释至0.2mM，然后进行4倍连续梯度稀释，共8个浓度梯度。用封板膜将96孔板密封，然后置于室温摇床孵育1小时。同时用浸板缓冲液(0.3％PEI(聚乙烯亚胺，购自：sigmaaldrich(西格玛奥德里奇)，型号：P314)，4℃保存)浸泡GF/C过滤板，至少0.5小时。结合孵育完毕用细胞收集器收集到GF/C过滤板上，用洗板缓冲液(50mM Tris-HCl,pH7.4，4℃保存)洗4次。50℃烘箱干燥1小时后把干燥好的GF/C过滤板底部封膜，用液体闪烁计数法检测滤膜残留放射性，每孔加入50μL闪烁液，并密封，使用Microbeta2(微孔板计数器，购自：PerkinElmer(珀金埃尔默仪器)，型号：CNLL0153)读数。计算待测样品对3H-flunitrazepam与GABAA受体膜蛋白结合的抑制活性，通过量效曲线拟合(GraphPad Prism 5软件)计算各待测样品的IC50，并通过IC50计算样品的Ki，从而评估化合物与α2-GABAA受体BZD位点的结合能力。
通过上述测定化合物与表达人源α2-GABAAR-HEK293细胞膜蛋白BZD位点结合亲和力的方法获得的代表性检测结果如表1所示。
本申请化合物对不同亚型GABAA受体的功能活性
通过电生理的方法检测待测药物对α1-GABAA受体和α2-GABAA受体的正向调控活性。具体方法如下所示：
化合物浓度设定：化合物筛选使用的化合物终浓度均为100nM。对于表达α1-GABAA受体的细胞系，GABA浓度为0.05～0.07μM(约EC2～4)；对于表达α2-GABAA受体的细胞系，GABA浓度为0.10～0.11μM(约EC7～8)。电生理试验采用全细胞膜片钳技术,该方法可参照文献(Nickolls,S.A.,et al.British Journal of Pharmacology,2018,175:708–725)报道的方法。电生理用细胞外液(ECS)成分如下：150mM NaCl，5mM KCl，2.5mM CaCl2，1mM MgCl2，10mM HEPES和10mM glucose(pH7.4)；电生理用电极内液(ICS)配方如下：140mM CsCl，11mM EGTA，10mM HEPES，2mM CaCl2，1mM MgCl2，4mM MgATP，2mM TEA(pH 7.3)。将GABA(γ-氨基丁酸)粉末用纯水配制成母液，在稀释于ECS中；化合物先用二甲基亚砜配制成为4mM的母液，再逐步稀释相应浓度的GABA-ECS中。溶液均在电生理测试前新鲜配制。
电生理信号采集使用EPC 10放大器及PatchMaster软件(HEKA)或Axon700B放大器及Clampex软件(AXON)。记录电极使用硼硅酸盐(borosilicate)玻璃拉制，电极电阻为4～6MΩ。胞外给药采用ALA-VC-8PGTM系统。选取单个独立生长的细胞，玻璃电极与细胞形成良好封接后破膜，形成全细胞模式。将细胞膜电位被钳制在-60mV，Gap-free模式下记录。试验时，先在胞外施加约20秒的细胞外液。待基线(Iprebaseline)稳定后，将胞外液切换至GABA-ECS。此时，可以检测到GABA引起的电流(Igaba)。大约10～20秒，待电流稳定后，将胞外液切换至化合物和GABA-ECS的混合溶液，直到此时可以检测到化合物和GABA共同引起的电流(Itreatment)。最后，将溶液切换至细胞外液，记录20～40秒终止试验。只有基线平稳、对照电流大小(Igaba-Iprebaseline)绝对值大于40pA且10～20秒内电流相对平稳的细胞才会用于化合物测试。
α1-GABAAR和α2-GABAAR的正向别构调节活性的实验结果分析采用PatchMaster v2x90.1或PatchMaster v2x90.3软件(EPC 10放大器)和Clampex10.6软件(Axon700B放大器)。各阶段的电流值按相应条件下稳定后电流的平均值计算。我们将对照电流定义为Igaba-Iprebaseline，化合物处理后电流定义为Itreatment-Iprebaseline。化合物的别构调节活性以百分比表示，按以下公式计算：功能活性＝[(Itreatment-Igaba)/(Igaba-Iprebaseline)]×100％。如果数值为负，则表示测试化合物对GABA受体的调节作用为反向别构调节。如果数值为正，则表示测试化合物对GABA受体的调节作用为正向别构调节。为了方便比较，以阳性化合物CVL-865作为参照物，进行归一化，即正向别构调节活性＝[化合物的功 能活性/阳性化合物的功能活性]×100％。化合物对α1-GABAA受体和α2-GABAA受体的正向别构调节活性测试结果如表1所示。
体外微核试验
1、遗传毒性研究指导原则
本实验设计基于实验目的，遵循下列实验设计指导原则：
(1)2016年修订的OECD化学品测试准则487，体外哺乳动物细胞微核试验；
(2)2011年11月生效的ICH S2(R1)：人用药物遗传毒性试验和结果分析指导原则。
2、材料和方法
实验系统和选择理由：本实验使用中国仓鼠卵巢细胞(CHO-WBL细胞)作为实验系统，该细胞系核型含有21条染色体，生长周期为12到13小时。CHO-WBL细胞最初来源于美国三藩市加利福利亚大学的S.Wolff博士，后在纽约哥伦比亚大学A.Bloom博士的实验室克隆，后被马里兰州Litton Bionetics实验室S.Galloway博士再次克隆。细胞储备液储存在液氮中。每一批次的细胞储备液都鉴定了细胞核型，并经检测证明未被支原体污染。细胞克隆后传代数将不会超过15次。之所以选择中国仓鼠卵巢细胞，是因为在OECD化学品测试指导原则487中推荐使用该细胞系，并被证明对许多种致染色体断裂剂/非整倍体诱变剂敏感(Marilyn J.et al.,2006)。
培养基和细胞生长条件：中国仓鼠卵巢细胞培养在McCoy’s 5A全培养基中(McCoy’s 5A全培养基补充加入10vt％胎牛血清、2mM GlutaMAXTM培养基、100units/mL青霉素和100μg/mL链霉素)，在标准环境中生长(温度37℃、CO2浓度为5％的高湿度培养箱)。给药前一天，将处于指数生长期的CHO-WBL细胞按照8 x 103个细胞/400μL McCoy’s 5A全培养基/孔接种到八孔细胞培养玻片上。
代谢活化系统：体外代谢活化系统(Maron and Ames,1983)包含β-萘黄酮和苯巴比妥诱导的大鼠肝脏微粒体匀浆(简称S9)和一个NADPH生成系统(即NADP二钠加异柠檬酸三钠)。S9从齐氏生物科技有限公司购买，使用前保存在-75℃冰箱里。S9匀浆的制备方法：将β-萘黄酮和苯巴比妥单次腹腔注射雄性Sprague Dawley大鼠，取材制备。
S9代谢活化系统的制备：先将NADP二钠盐和异柠檬酸三钠盐溶解在不含血清培养基中，制成核心溶液，并用0.22μm的滤头过滤除菌。临用前，将S9匀浆融化，将S9代谢活化系统的各组分按照如下表2所示比例混匀。
表2 S9代谢活化系统组分及配比
剂量探索实验：在剂量探索实验中，每个给药处理系列中设立8个给药浓度。受试物每个浓度设1孔细胞。在ICH指导原则S2(R1)中，当不受限于受试物在溶剂/培养基中的溶解度或受试物的细胞毒性时，推荐的最高浓度为1mM或0.5mg/mL，取较低者。然而，若受限于溶解度，测试的最高浓度为在显微镜下培养基中可见少量沉淀的最低浓度。为了确保能够达到在培养基中的溶解度上限，可以测试多达4个含有可见沉淀的浓度。
给药处理：S9代谢活化3小时给药系列中，将每孔中培养基吸掉，参照ICH S2(R1)指导原则，添加含有合适浓度受试物/对照品的不含血清McCoy’s 5A/S9混合物培养基。
非代谢活化3小时给药系列中，将每孔中培养基吸掉，添加含有合适浓度受试物/对照品的McCoy’s 5A全培养基。
将细胞放在标准条件下孵育3小时。3小时后，将各孔中旧培养基吸掉，用McCoy’s 5A全培养基洗2次。每孔加入含有细胞松弛素B(培养基中终浓度为约3μg/mL)的McCoy’s 5A全培养基，接着培养21小时直至收获。
对于非代谢活化24小时给药系列，处理方式与非代谢活化3小时给药系列类似，不同之处在于细胞将与受试物/对照品在含有细胞松弛素B(培养基中终浓度为约3μg/mL)的全培养基中培养 24小时(±30分钟)。
玻片制备：收获时，倒掉八孔玻片中的培养基，用吸水纸吸干，每孔中加入75mM KCl低渗液，然后将细胞连续更换两次固定液(无水甲醇：冰醋酸，3:1v/v)固定5分钟，风干。避光条件下，用吖啶橙染色并风干。
计数细胞毒性：通过低倍镜检查呈现出来的染色情况，评估每张玻片的所有孔的可被分析的可能性(足够数量的细胞)。在荧光显微镜下，每孔分析至少500个细胞，分别计数含有的单核、双核和多核细胞的数目，以计算胞质分裂阻断增殖指数(CBPI)。
用CBPI评估受试物的细胞毒性。CBPI的计算公式如下：
％细胞毒性＝100-100{(CBPIT-1)/(CBPIC-1)}
其中，T＝受试物处理培养组，C＝溶媒对照培养组。
当CBPI为1时，等于100％的细胞毒性。
微核主实验：微核主实验将CHO-WBL细胞暴露于受试物的3-8个浓度下，同时设立与溶剂组相同处理的阳性对照组(环磷酰胺一水合物)和溶媒对照组(二甲基亚砜)，每个剂量组均设2孔细胞。在非活化测试系统中，给药时间为3和24小时，在S9活化测试系统中，受试物暴露时间为3小时。受试物测试的上限取决于它在培养基中的溶解度，但总的来说不会超过最大浓度1mM或0.5mg/mL，取较低者。选择微核频率分析的最高剂量时，该受试物剂量组与相应溶媒对照组相比细胞毒性应不超过50％太多。若最高浓度不受限于细胞毒性，为了达到培养基中溶解度上限，可以测试多个含有可见沉淀的剂量，且最高剂量组的培养基中可见少量可识别的沉淀。
选择微核分析的剂量：CHO-WBL细胞微核分析剂量的选择将取决于受试物的细胞毒性。选择用于评估的最高剂量将诱导不超过50％太多的细胞毒性。至少再分析两个更低的剂量组(在中等细胞毒性到非细胞毒性范围内，如适用)。若受试物没有细胞毒性，则选择在给药末期在显微镜下可以观察到沉淀的剂量，或者最大待测浓度(1mM或0.5mg/mL，取较低者)为用于微核分析的最高浓度。
计数微核频率：所有选择作微核分析的玻片均用随机生成的盲码标注，以减少读片的主观性，盲码由不参与读片的人员制作。盲码标记的片子标签中包含以下信息：实验编号、处理系列和随机数字。
每一剂量组分析2000个双核细胞(每孔1000个)，以计数含有微核的双核细胞频率。
双核细胞应符合下列标准才可计数：
a.具有完整的细胞膜。b.两个子核大小相等。
微核的判断标准如下：
a.微核具有与主核类似的荧光强度。b.微核应游离于细胞质中。c.微核边缘光滑，直径小于等于主核直径的三分之一。
玻片在签署实验报告前废弃。
数据
数据报告：给药组的细胞毒性根据与对应的溶媒对照相比的细胞毒性来判定并作呈列。微核主实验中每一孔和每一剂量组的微核频率也作呈列。
统计分析：使用Fisher’s确切概率法比较给药组与相应溶媒对照组含有微核的双核细胞发生频率差异的显著性。作多重比较时，P值经过Bonferroni校正。多组数据比较时，用Cochran-Armitage检验来检测剂量效应关系。
当P≤0.05时，可认为差异具有显著性。
实验有效性标准：一个可接受的遗传毒性测试必须满足以下标准：溶媒/阴性对照微核细胞的频率必须与历史阴性对照数据具有可比性。应该至少有三个可被分析的浓度。阳性对照微核细胞的比例应该和平行溶媒/阴性对照组相比有统计学上的显著的升高(p≤0.05)。
实验结果的评判标准：在该实验有效前提下，根据指导原则ICH S2(R1)，试验结果判定，需结合不同给药处理条件下(加S9给药3小时，不加S9给药3小时，不加S9给药24小时)，不同 受试物浓度下，微核率的结果，参照试验结果的评判标准，综合分析判定受试物的体外微核结果为阴性，阳性或可疑阳性，结果以阴性表示为佳。具体标准评估实验结果如下：
阳性：若受试物同时满足以下标准，可认为该受试物为阳性：在一个或多个剂量组中观察到微核细胞频率与平行溶媒对照组相比显著性增加。微核频率的增加具有剂量效应关系。在一个或多个剂量组中观察到微核细胞频率超出实验室阴性历史数据范围。
阴性：若以上三点均不符合，可认为该受试物为阴性。
可疑阳性：若一个受试物仅部分满足以上三点，应科学地判断。浓度相关效应的迹象被认为是有用的，但在评价阳性结果时并不是必要的。将考虑生物相关性，例如浓度范围内和浓度之间的反应一致性，以及(如适用)实验之间的反应一致性，或仅在高或极强细胞毒性浓度下发生的效应。
本申请各化合物对α2-GABAA受体的亲和活性以及对α1、α2-GABAA受体的正向别构调节活性结果见下表3，体外微核测试的结果如表4所示。
需要注意的是，由于本申请所述化合物的优势在于受体较高的选择性而不仅仅在于药物的绝对活性，同时考虑到在不同的检测体系下，化合物的功能活性绝对值会有一定差异，不具有可比性，因此，为了方便比较，本申请实验在同一检测系统下同步检测对照和涉及的化合物，并以阳性化合物CVL-865作为参照物，进行归一化处理，即正向别构调节活性＝[化合物的功能活性/阳性化合物的功能活性]×100％，具体结果见下表3。
表3部分化合物对α2-GABAA受体的亲和活性及对α1、α2-GABAA受体的正向别构调节活性
本申请实施例化合物对比参考化合物CVL-865，选择性更好，在保持适当的α2-GABAA受体亲和力和电生理活性的情况下，使α1-GABAA受体电生理活性小于CVL-865的70％，这样在维持体内药效的同时，有更小的副作用。
表4体外微核测试的结果
由体外微核结果可见，本申请化合物具有更好的遗传毒安全性，有更好的成药性。
显然，上述实施例仅仅是为清楚地说明所作的举例，而并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。

Claims (18)

  1. 一种式(1)所示化合物及其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、同位素体、多晶型物或溶剂合物：
    其中，
    R1为取代或者未取代的杂环与杂环形成的稠合基；
    R2选自H、卤素、OH、C1-C6烷氧基或CN；
    R3选自H、取代或者未取代的直链或者支链C1-C6烷基或取代或者未取代的C3-C6环烷基。
  2. 如权利要求1所述的化合物，其特征在于，形成所述R1所示稠合基的两个杂环彼此独立地为含有1-3个选自N、O或S的环杂原子的5-7元饱和或不饱和的单环基。
  3. 如权利要求1或2所述的化合物，其特征在于，所述R1具有式(2)、(3)、(4)、(5)、(6)或(7)所示的结构：
    其中，
    所述R4选自氢、C1-C6烷基、C3-C6环烷基、C1-C6烷氧基、C1-C6烷硫基或者C1-C6烷基磺酰基；上述基团可选择的未被取代或者彼此独立的被1-4个分别选自卤素、羟基、C1-C3烷基、卤代C1-C3烷基、C1-C3烷氧基或者卤代C1-C3烷氧基中的至少一种所取代；
    所述R5选自氢、卤素、羟基、氧代、C1-C6烷基酰基、C1-C6烷基酰胺基、C1-C6烷氧基亚胺基、C1-C6烷基、C2-C6烯基、C2-C6炔基、C3-C6环烷基、C1-C6烷氧基、C6-C10的芳基、含有 1-3个杂原子的5-10元的杂芳基、含有1-3个杂原子的C1-C6的非芳族杂环；上述基团可选择的未被取代或者彼此独立的被1-4个分别选自卤素、羟基、C1-C3烷基、卤素取代的C1-C3烷基、C1-C3烷氧基、卤代C1-C3烷氧基或者含有1-3个N或O的杂原子的3-7元杂环烷基中的至少一种所取代；
    所述A环是指具有含N的5-7元饱和或部分不饱和的单环；m为1、2或3，n为1或2，表示连接位点。
  4. 根据权利要求3所述的化合物，其特征在于，所述R4选自氢、甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、乙烯基、乙炔基、环丙基、环丁基、甲氧基、乙氧基、正丙氧基、-CH2-O-CH3、哌啶、甲基肟、甲基磺酰基或者乙基磺酰基；上述基团可选择的未被取代或者彼此独立的被C1-C3烷氧基或卤素取代。
  5. 根据权利要求3或4所述的化合物，其特征在于，所述R5选自氢、F、Cl、Br、羟基、甲酰基、乙酰基、甲酰胺基、乙酰胺基、甲氧亚胺基、乙氧亚胺基、甲基、乙基、异丙基、环丙基、环丁基、环戊基、环己基、甲氧基、乙氧基、苯基、苯甲基、四氢呋喃基、四氢吡喃基、六氢吡啶基、氧杂环丁烷、氮杂环丁烷；上述基团可选择的未被取代或者彼此独立的被1-4个分别选自F、Cl、Br、羟基、甲氧基、甲基、四氢吡咯、吗啉基或者-CH2-CF3中的至少一种所取代。
  6. 根据权利要求5所述的化合物，其特征在于，所述R5选自H、甲基、乙基、异丙基、 环丙基、氟代环丙基、 或者
  7. 如权利要求3-6任一项所述的化合物，其特征在于，所述A环为含有1个、2个或3个N的环杂原子的5-6元饱和或不饱和的单环基；其中，
    当R1具有式(2)或(7)所示的结构时，所述A环具有如下结构：
    或者
    当R1具有式(3)、(4)、(5)或(6)所示的结构时，A环具有如下结构：
    或者
    其中，表示连接位点。
  8. 如权利要求3-7任一项所述的化合物，其特征在于，所述R1为取代或者未取代的吡啶稠杂环。
  9. 如权利要求8所述的化合物，其特征在于，所述R1为取代或者未取代的吡啶并三氮唑或者取 代或者未取代的吡啶并咪唑。
  10. 如权利要求3-7任一项所述的化合物，其特征在于，所述R1选自如下化合物中的任意一种：
    或者
  11. 如权利要求3-7任一项所述的化合物，其特征在于，所述R1选自如下结构中的任意一种：
    或者并且，
    所述R4选自甲氧基C1-C3烷基或C1-C3烷氧基；优选为甲氧基甲基或者甲氧基；
    所述R5选自H、甲基、乙基、异丙基、环丙基、氟代环丙基、
  12. 如权利要求1-11任一项所述的化合物，其特征在于：
    所述R2选自H或F；
    所述R3选自H或直链或者支链C1-C6烷基。
  13. 如权利要求1-12任一项所述的化合物，其特征在于，所述化合物选自以下化合物中的任意一种：



  14. 一种药物组合物，其特征在于，其包含如权利要求1-13中任一项所述的化合物或其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、同位素体、多晶型物或溶剂合物中的至少一种，并可选择的添加药学上可接受的载体和/或辅助剂。
  15. 如权利要求1-13中任一项所述的化合物或其立体异构体、互变异构体、前体药物、药学上可接受的盐、无定形物、同位素体、多晶型物或溶剂合物，或权利要求10所述的药物组合物用于制备治疗或预防与GABAA受体有关的疾病的药物中的用途。
  16. 根据权利要求15所述的用途，其特征在于，所述与GABAA受体有关的疾病选自如下至少一种：疼痛、阿尔茨海默氏病、多梗塞性痴呆、癫痫、瘙痒或中风。
  17. 如权利要求16所述的用途，其特征在于，所述的疼痛包括神经病理性疼痛、炎性疼痛和癌性痛。
  18. 如权利要求16或17所述的用途，其特征在于，所述的疼痛选自：头痛，面部痛、颈痛、肩痛、背痛、胸痛、腹痛、背部痛、腰痛、下肢痛、肌肉与骨骼疼痛、血管疼痛、痛风、关节炎疼痛、内脏疼痛、感染性疾病导致的疼痛、多骨疼痛、镰刀细胞贫血、自身免疫性疾病、多发性硬化或炎症有关的疼痛，损伤或手术引起的慢性疼痛、伤害感受性疼痛、疼痛性糖尿病、三叉神经痛、腰部或子宫颈神经根病痛、舌咽神经痛、自主神经反射性疼痛、反射性交感神经营养不良、神经根撕脱、癌症、化学损伤、毒素、营养缺乏、病毒或细菌感染或退行性骨关节病有关的疼痛。
PCT/CN2023/076805 2022-02-25 2023-02-17 咪唑并哒嗪类衍生物、其制备方法、药物组合物和用途 WO2023160475A1 (zh)

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Citations (4)

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Publication number Priority date Publication date Assignee Title
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CN104837843A (zh) * 2012-12-14 2015-08-12 辉瑞有限公司 作为gabaa受体调节剂的咪唑并哒嗪衍生物
WO2015189744A1 (en) * 2014-06-12 2015-12-17 Pfizer Limited Imidazopyridazine derivatives as modulators of the gabaa receptor activity.
WO2017098367A1 (en) * 2015-12-10 2017-06-15 Pfizer Limited 4-(biphen-3-yl)-1h-pyrazolo[3,4-c]pyridazine derivatives of formula (i) as gaba receptor modulators for use in the treatment of epilepsy and pain

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Publication number Priority date Publication date Assignee Title
CN101384559A (zh) * 2006-03-24 2009-03-11 神经研究公司 苯并咪唑衍生物及其在调节gabaa受体复合物中的应用
CN104837843A (zh) * 2012-12-14 2015-08-12 辉瑞有限公司 作为gabaa受体调节剂的咪唑并哒嗪衍生物
WO2015189744A1 (en) * 2014-06-12 2015-12-17 Pfizer Limited Imidazopyridazine derivatives as modulators of the gabaa receptor activity.
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