WO2022100614A1 - 苯并脲环衍生物及其制备方法和应用 - Google Patents

苯并脲环衍生物及其制备方法和应用 Download PDF

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WO2022100614A1
WO2022100614A1 PCT/CN2021/129783 CN2021129783W WO2022100614A1 WO 2022100614 A1 WO2022100614 A1 WO 2022100614A1 CN 2021129783 W CN2021129783 W CN 2021129783W WO 2022100614 A1 WO2022100614 A1 WO 2022100614A1
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compound
reaction solution
mmol
pharmaceutically acceptable
acceptable salt
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French (fr)
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吴凌云
魏霞蔚
肖哲明
黎健
陈曙辉
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Medshine Discovery Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/416—1,2-Diazoles condensed with carbocyclic ring systems, e.g. indazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
    • C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond

Definitions

  • the invention relates to the field of medicine, in particular to a class of benzourea ring derivatives and a preparation method and application thereof, in particular to a compound represented by formula (I) and a pharmaceutically acceptable salt thereof.
  • RHO-associated protein kinase a serine/threonine protein kinase
  • ROCK RHO-related protein kinase
  • MLC myosin light chain
  • ROCK kinase is involved in the regulation of the immune response of TH17 cells and the activation of fibroblasts, which can expand adaptation It is used for pulmonary fibrosis, asthma and other lung diseases, and further indications include autoimmune diseases.
  • the ROCK kinase family includes two isoforms, ROCK1 and ROCK2.
  • ROCK2 kinase is associated with inflammation and fibrosis. Selective ROCK2 inhibitors do not induce vasodilation at high concentrations in isolated vasodilation experiments, which can reduce cardiovascular side effects.
  • ROCK1 knockout mice have low embryonic mortality, most of them die after birth due to cytoskeletal variation caused by reduced MLC phosphorylation, while 90% of ROCK2 knockout mice die in the embryonic stage, but surviving mice and wild-type mice There was no difference, and selective inhibition of ROCK2 activity may have a higher safety profile. Therefore, selective ROCK2 protein kinase inhibitors can avoid the cardiovascular side effects of the drug.
  • KD025 (WO2006105081; WO2008054599; WO2010104851; WO2014055996) is an oral ROCK2 kinase selective inhibitor developed by Kadmon Company. Studies have shown that KD025 represents a novel mechanism for the treatment of idiopathic pulmonary fibrosis (IPF) by inhibiting fibrosis-regulating proteins such as RHO kinase. Idiopathic pulmonary fibrosis (IPF) may be caused by physical damage.
  • IPF idiopathic pulmonary fibrosis
  • the body's response to injury involves the reorganization of the actin cytoskeleton of various cells (such as epithelial cells, fibroblasts, endothelial cells, and macrophages), and the assembly of actin filaments and actomyosin
  • actomyosin The contraction of actomyosin is guided and regulated by RHO kinase family proteins (including ROCK1 and ROCK2).
  • RHO kinase family proteins include ROCK1 and ROCK2
  • Previous studies have shown that RHO kinase family proteins are activated in the lungs of IPF patients and animal models of the disease, and RHO kinase inhibitors can prevent tissue fibrosis in these models and induce regression of established fibrosis . It has completed a Phase II clinical trial for the treatment of moderate to severe psoriasis and is in the Phase II clinical study phase for the treatment of idiopathic pulmonary fibrosis (IPF).
  • IPPF idiopathic pulmonary
  • WO2014134388 and WO2016028971 also disclose a class of compounds whose general structural formulas are shown in formula (a) and (b), which can also be used for cardiovascular diseases, neuropathological diseases, tumors, autoimmune diseases, and pulmonary fibrosis. , treatment of inflammatory diseases, etc.
  • the compounds of the present invention are ROCK2 inhibitors, which have significant kinase inhibitory activity.
  • the present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
  • T 1 is N or CH;
  • T 3 is N or CR 3 ;
  • T 4 is N or CH
  • R 1 , R 2 , R 3 and R 4 are each independently H, F, Cl, Br, I, -OH, -CN, -NH 2 , -NO 2 , C 1-3 alkyl or C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy groups are optionally 1, 2 or 3 independently selected from F, Cl, Br, I, -OH, -OCH 3 , -CN , -NH 2 or -NO 2 substituents;
  • R 6 and R 7 are each independently H, F, Cl or C 1-3 alkyl, wherein said C 1-3 alkyl is optionally selected from 1, 2 or 3 independently selected from F, Cl, Br, I , -OH, -OCH 3 , -CN, -NH 2 or -NO 2 substituent;
  • Each R 8 is independently H, F, Cl, Br, I, -OH, -CN, -NH 2 , -NO 2 , C 1-3 alkyl or C 1-3 alkoxy, wherein said C 1 -3 alkyl and C 1-3 alkoxy optionally by 1, 2 or 3 independently selected from F, Cl, Br, I, -OH, -OCH 3 , -CN, -NH 2 or -NO 2 Substituent substituted; n is 1, 2, 3 or 4.
  • the above-mentioned compound or a pharmaceutically acceptable salt thereof has the structure represented by formula (I-1) or (I-2):
  • R 1 , R 2 , R 3 , R 4 , R 6 , R 7 , R 8 , T 1 , T 3 , T 4 and n are as defined in the present invention.
  • R 1 , R 2 , R 3 and R 4 are each independently H, F, Cl, Br and -CH 3 , and other variables are as defined herein.
  • R 6 and R 7 are each independently H and -CH 3 , wherein the -CH 3 is optionally 1, 2 or 3 independently selected from F, Cl, Br, -OH or -NH 2 substituents, other variables are as defined in the present invention.
  • R6 and R7 above are each independently H and -CH2NH2 , and other variables are as defined herein .
  • R 8 is independently H, F, Cl, Br, -OH, -CN, -NH 2 , -NO 2 or -OCH 3 , and other variables are as defined herein.
  • n, R8 and other variables are as defined in the present invention.
  • the above-mentioned compounds or pharmaceutically acceptable salts thereof have the structures represented by formulae (I-3) to (I-7):
  • R 1 , R 2 , R 3 , R 4 , R 6 , R 7 , R 8 and n are as defined in the present invention.
  • the above-mentioned compounds or pharmaceutically acceptable salts thereof have the structures represented by formulae (I-8) to (I-12):
  • R 1 , R 6 , R 7 , R 8 and n are as defined in the present invention.
  • the above-mentioned compounds or pharmaceutically acceptable salts thereof have the structures represented by formulae (I-13) to (I-17):
  • R 1 , R 8 and n are as defined in the present invention; the carbon atom with "*" is a chiral carbon atom, in the form of (R) or (S) single enantiomer or enriched in one enantiomer form exist.
  • the above compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:
  • the compounds of the present invention have high kinase inhibitory activity against ROCK2.
  • the term "pharmaceutically acceptable” refers to those compounds, materials, compositions and/or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue , without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • salts refers to salts of the compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases.
  • base addition salts can be obtained by contacting the neutral forms of such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts.
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent.
  • Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts including, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, and methanesulfonic acids; also include salts of amino acids such as arginine, etc. , and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and thus can be converted into either base
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the acid or base containing parent compound by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.
  • the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and racemic mixtures thereof and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this within the scope of the invention.
  • Additional asymmetric carbon atoms may be present in substituents such as alkyl. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.
  • enantiomers or “optical isomers” refer to stereoisomers that are mirror images of each other.
  • cis-trans isomer or “geometric isomer” result from the inability to rotate freely due to double bonds or single bonds to ring carbon atoms.
  • diastereomer refers to a stereoisomer in which the molecule has two or more chiral centers and the molecules are in a non-mirror-image relationship.
  • tautomer or “tautomeric form” refers to isomers of different functional groups that are in dynamic equilibrium and are rapidly interconverted at room temperature.
  • a chemical equilibrium of tautomers can be achieved if tautomers are possible (eg, in solution).
  • proton tautomers also called prototropic tautomers
  • prototropic tautomers include interconversions by migration of protons, such as keto-enol isomerization and imine-ene Amine isomerization.
  • Valence tautomers include interconversions by recombination of some bonding electrons.
  • keto-enol tautomerization is the interconversion between two tautomers, pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
  • the terms “enriched in one isomer”, “enriched in isomers”, “enriched in one enantiomer” or “enriched in one enantiomer” refer to one of the isomers or pairs
  • the enantiomer content is less than 100%, and the isomer or enantiomer content is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or Greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
  • isomeric excess or “enantiomeric excess” refer to the difference between two isomers or relative percentages of two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80% .
  • Optically active (R)- and (S)-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure desired enantiomer.
  • a diastereomeric salt is formed with an appropriate optically active acid or base, followed by conventional methods known in the art
  • the diastereoisomers were resolved and the pure enantiomers recovered.
  • separation of enantiomers and diastereomers is usually accomplished by the use of chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (eg, from amines to amino groups) formate).
  • Compounds of the invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound.
  • compounds can be labeled with radioisotopes, such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C).
  • deuterated drugs can be formed by replacing hydrogen with deuterium, and the bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All transformations of the isotopic composition of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.
  • substituted means that any one or more hydrogen atoms on a specified atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence of the specified atom is normal and the substituted compound is stable.
  • oxygen it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on aromatic groups.
  • optionally substituted means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be arbitrary on a chemically achievable basis.
  • any variable eg, R
  • its definition in each case is independent.
  • the group may optionally be substituted with up to two Rs, with independent options for R in each case.
  • combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
  • linking group When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
  • substituents When a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A.
  • substituents do not specify through which atom it is attached to the substituted group, such substituents may be bonded through any of its atoms, for example, pyridyl as a substituent may be through any one of the pyridine rings. The carbon atom is attached to the substituted group.
  • the direction of attachment is arbitrary, for example,
  • the linking group L in the middle is -MW-, at this time -MW- can connect ring A and ring B in the same direction as the reading order from left to right. It is also possible to connect ring A and ring B in the opposite direction to the reading order from left to right.
  • Combinations of the linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
  • any one or more sites in the group can be linked to other groups by chemical bonds.
  • connection method of the chemical bond is not located, and there is an H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will decrease correspondingly with the number of the connected chemical bond. the group.
  • the chemical bond connecting the site to other groups can be represented by straight solid line bonds straight dotted key or wavy lines express.
  • a straight solid bond in -OCH 3 indicates that it is connected to other groups through the oxygen atom in this group;
  • the straight dashed bond in the group indicates that it is connected to other groups through the two ends of the nitrogen atom in the group;
  • the wavy lines in the phenyl group indicate connections to other groups through the 1 and 2 carbon atoms in the phenyl group.
  • C 1-3 alkyl is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms.
  • the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (eg methyl), divalent (eg methylene) or multivalent (eg methine) .
  • Examples of C1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.
  • C1-3alkoxy refers to those alkyl groups containing 1 to 3 carbon atoms attached to the remainder of the molecule through an oxygen atom.
  • the C 1-3 alkoxy group includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy and the like.
  • Examples of C 1-3 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
  • Cn-n+m or Cn - Cn+m includes any particular instance of n to n+ m carbons, eg C1-12 includes C1 , C2 , C3, C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , also including any range from n to n+ m , eg C 1-12 includes C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 , etc.; in the same way, n yuan to n +m-membered means that the number of atoms in the ring is from n to n+m, for example, 3-12-membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membere
  • leaving group refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (eg, affinity substitution reaction).
  • representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonic acid Esters, etc.; acyloxy, such as acetoxy, trifluoroacetoxy, and the like.
  • protecting group includes, but is not limited to, "amino protecting group", “hydroxy protecting group” or “thiol protecting group”.
  • amino protecting group refers to a protecting group suitable for preventing side reactions at the amino nitrogen position.
  • Representative amino protecting groups include, but are not limited to: formyl; acyl groups, such as alkanoyl groups (eg, acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl groups, such as tert-butoxycarbonyl (Boc) ; Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); Arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-di -(4'-Methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS), 2-(trimethylsilyl (TMS),
  • hydroxy protecting group refers to a protecting group suitable for preventing hydroxyl side reactions.
  • Representative hydroxy protecting groups include, but are not limited to: alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (eg acetyl); arylmethyl groups such as benzyl (Bn), p-methyl Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and the like.
  • alkyl groups such as methyl, ethyl and tert-butyl
  • acyl groups such as alkanoyl (eg acetyl)
  • arylmethyl groups such as benzyl (Bn), p-methyl Oxybenzyl (PMB), 9-fluorenyl
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.
  • the structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction method (SXRD), the cultured single crystal is collected by Bruker D8 venture diffractometer, the light source is CuK ⁇ radiation, and the scanning mode is: After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.
  • SXRD single crystal X-ray diffraction method
  • the cultured single crystal is collected by Bruker D8 venture diffractometer
  • the light source is CuK ⁇ radiation
  • the scanning mode is: After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.
  • the solvent used in the present invention is commercially available.
  • g stands for grams; mg stands for milligrams; ⁇ L stands for microliters; mL stands for milliliters; mol stands for moles; mmol stands for millimoles; /L; ⁇ M stands for micromoles/liter; nM stands for nanomoles/liter.
  • Me represents methyl; Boc represents tert-butoxycarbonyl; DMSO represents dimethyl sulfoxide; DMSO-d 6 represents deuterated dimethyl sulfoxide.
  • Urotropine (30.6 g, 218 mmol) was added to a solution of compound B-1 (50.0 g, 218 mmol) in dichloromethane (625 mL), and the reaction solution was stirred at 30 degrees Celsius for 1 hour. The reaction solution was filtered, and the filter cake was collected to obtain the crude compound B-2.
  • Urotropine (2.68 g, 19.1 mmol) was added to a solution of compound 1-2 (4.40 g, 19.1 mmol) in dichloromethane (60 mL), and the reaction solution was stirred at 20 degrees Celsius for 1 hour. The reaction solution was filtered, and the filter cake was collected to obtain the crude compound 1-3.
  • the hydrochloride salt of compound 1-4 (2.80 g, 13.8 mmol) was dissolved in water (25 mL) and tetrahydrofuran (25 mL), then di-tert-butyl dicarbonate (2.41 g, 11.1 mmol, 2.54 mL) and hydrogen carbonate were added Sodium (3.58 g, 41.5 mmol).
  • the reaction solution was stirred at 20°C for 1 hour.
  • the reaction solution was diluted with water (60 mL) and extracted with ethyl acetate (100 mL x 2).
  • the combined organic phases were washed once with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure.
  • the crude product concentrated under reduced pressure was separated and purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate, 30/1 ⁇ 10/1, V/V) to obtain compound 1-5.
  • the fifth step of reference example 2 obtains the crude product, and the obtained crude product is subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150 x 30 mm x 4 ⁇ m; mobile phase: [0.05% aqueous hydrochloric acid solution-acetonitrile]; B%: 7%-27 %, 11 minutes) separation and purification to obtain the hydrochloride of compound 3.
  • Assay buffer solution 20 mM 4-hydroxyethylpiperazineethanesulfonic acid (pH 7.5), 10 mM magnesium chloride, 1 mM ethylene glycol diethyl ether diamine tetraacetic acid, 0.02% polyoxyethylene lauryl ether, 0.02 mg/mL bovine serum albumin , 0.1 mM sodium vanadate, 2 mM dithiothreitol, 1% DMSO.
  • the protein kinase inhibitory activity of the compounds was expressed as the residual protein kinase activity relative to the blank substrate (DMSO alone). IC50 values and curves were calculated using the Prism software package (GraphPad Software, San Diego California, USA).
  • the compounds of the present invention have significant and even unexpected ROCK2 inhibitory activity.

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Abstract

式(I)所示的苯并脲环衍生物或其药学上可接受的盐以及其制备方法和应用。

Description

苯并脲环衍生物及其制备方法和应用
本申请主张如下优先权：
CN202011259175.6，2020年11月11日。
技术领域
本发明涉及药物领域，具体涉及一类苯并脲环衍生物及其制备方法和应用，具体涉及式(I)所示化合物及其药学上可接受的盐。
背景技术
RHO相关蛋白激酶(Rho associated kinase,简称ROCK)，属于丝氨酸/苏氨酸蛋白激酶，是RHO的下游靶效应分子，在人体内广泛表达。RHO相关蛋白激酶(ROCK)参与肌球蛋白轻链(MLC)的调节，适用于血管舒张的治疗，新的研究支持ROCK激酶参与TH17细胞的免疫反应的调控和纤维母细胞的活化，可以扩展适应症用于肺纤维化，哮喘等肺部疾病，进一步的适应症拓展包括自身免疫疾病。ROCK激酶家族包括ROCK1和ROCK2两个亚型，ROCK2激酶和炎症作用和纤维化作用相关，选择性ROCK2抑制剂在离体血管舒张实验中在高浓度也没有引起血管舒张，可以减少心血管副作用。ROCK1敲除小鼠虽然胚胎死亡率不高，但是出生后大部分死于MLC磷酸化降低造成的细胞骨架变异，而ROCK2敲除小鼠90％在胚胎期死亡，但是存活的小鼠和野生型没有区别，选择性抑制ROCK2的活性可能具有更高的安全性。因此，选择性ROCK2蛋白激酶抑制剂可以避免药物的心血管副作用。
KD025(WO2006105081；WO2008054599；WO2010104851；WO2014055996)是Kadmon公司开发的口服ROCK2激酶选择性的抑制剂。研究表明，KD025是通过抑制RHO激酶这类调节纤维化的蛋白来治疗特发性肺纤维化(IPF)这种新机制的代表。特发性肺纤维化(IPF)的诱因可能是肌体损伤。肌体对损伤的应答涉及多种细胞(比如上皮细胞、成纤维细胞、内皮细胞和巨噬细胞等)的肌动蛋白细胞骨架的重组，而肌动蛋白丝(actin filament)的组装和肌动球蛋白(actomyosin)的收缩则由RHO激酶家族蛋白质(包括ROCK1和ROCK2)指导调控。之前的研究显示，RHO激酶家族蛋白会在IPF患者以及这种疾病的动物模型肺部激活，而RHO激酶抑制剂可以预防这些模型中的组织纤维化过程，并且能诱导已经建立的纤维化出现消退。目前已完成中度至重度银屑病治疗的II期临床试验，并处于特发性肺纤维化(IPF)治疗的II期临床研究阶段。
Figure PCTCN2021129783-appb-000001
WO2014134388和WO2016028971也公开了一类化合物，其结构通式如式(a)和式(b)所示，这类化合物也可用于心血管疾病、神经病理学疾病、肿瘤、自免疫疾病、肺纤维化、炎症疾病等的治疗。
Figure PCTCN2021129783-appb-000002
本发明化合物为ROCK2抑制剂，其具有显著的激酶抑制活性。
发明内容
一方面，本发明提供了式(I)所示化合物或其药学上可接受的盐，
Figure PCTCN2021129783-appb-000003
其中，
T 1为N或CH；T 3为N或CR 3；
当T 2为O时，T 3为N，结构单元
Figure PCTCN2021129783-appb-000004
为
Figure PCTCN2021129783-appb-000005
当T 2为N时，结构单元
Figure PCTCN2021129783-appb-000006
为
Figure PCTCN2021129783-appb-000007
T 4为N或CH；
R 1、R 2、R 3和R 4各自独立地为H、F、Cl、Br、I、-OH、-CN、-NH 2、-NO 2、C 1-3烷基或C 1-3烷氧基，其中所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN、-NH 2或-NO 2的取代基所取代；
R 6和R 7各自独立地为H、F、Cl或C 1-3烷基，其中所述C 1-3烷基任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN、-NH 2或-NO 2的取代基所取代；
各R 8独立地为H、F、Cl、Br、I、-OH、-CN、-NH 2、-NO 2、C 1-3烷基或C 1-3烷氧基，其中所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN、-NH 2或-NO 2的取代基所取代；n为1、2、3或4。
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其化合物具有式(I-1)或(I-2)所示结构：
Figure PCTCN2021129783-appb-000008
其中，R 1、R 2、R 3、R 4、R 6、R 7、R 8、T 1、T 3、T 4和n如本发明所定义。
在本发明的一些方案中，上述R 1、R 2、R 3和R 4各自独立地为H、F、Cl、Br和-CH 3，其他变量如本 发明所定义。
在本发明的一些方案中，上述R 6和R 7各自独立地为H和-CH 3，其中所述-CH 3任选被1、2或3个独立选自F、Cl、Br、-OH或-NH 2的取代基所取代，其他变量如本发明所定义。
在本发明的一些方案中，上述R 6和R 7各自独立地为H和-CH 2NH 2，其他变量如本发明所定义。
在本发明的一些方案中，上述R 8独立地为H、F、Cl、Br、-OH、-CN、-NH 2、-NO 2或-OCH 3，其他变量如本发明所定义。
在本发明的一些方案中，上述结构单元
Figure PCTCN2021129783-appb-000009
为
Figure PCTCN2021129783-appb-000010
Figure PCTCN2021129783-appb-000011
n、R 8及其他变量如本发明所定义。
在本发明的一些方案中，上述结构单元
Figure PCTCN2021129783-appb-000012
为
Figure PCTCN2021129783-appb-000013
Figure PCTCN2021129783-appb-000014
R 8及其他变量如本发明所定义。
在本发明的一些方案中，上述结构单元
Figure PCTCN2021129783-appb-000015
为
Figure PCTCN2021129783-appb-000016
Figure PCTCN2021129783-appb-000017
其他变量如本发明所定义。
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其化合物具有式(I-3)～(I-7)所示结构：
Figure PCTCN2021129783-appb-000018
Figure PCTCN2021129783-appb-000019
其中，R 1、R 2、R 3、R 4、R 6、R 7、R 8和n如本发明所定义。
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其化合物具有式(I-8)～(I-12)所示结构：
Figure PCTCN2021129783-appb-000020
其中，R 1、R 6、R 7、R 8和n如本发明所定义。
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其化合物具有式(I-13)～(I-17)所示结构：
Figure PCTCN2021129783-appb-000021
Figure PCTCN2021129783-appb-000022
其中，R 1、R 8和n如本发明所定义；带“*”的碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在。
本发明还有一些方案是由上述变量任意组合而来。
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其中化合物选自：
Figure PCTCN2021129783-appb-000023
Figure PCTCN2021129783-appb-000024
技术效果
本发明化合物对ROCK2具有很高的激酶抑制活性。
定义和说明
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠 的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物的中性形式接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机胺或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物的中性形式接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，所述无机酸包括例如盐酸、氢溴酸、硝酸、碳酸，碳酸氢根，磷酸、磷酸一氢根、磷酸二氢根、硫酸、硫酸氢根、氢碘酸、亚磷酸等；以及有机酸盐，所述有机酸包括如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸和甲磺酸等类似的酸；还包括氨基酸(如精氨酸等)的盐，以及如葡糖醛酸等有机酸的盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。
除非另有说明，术语“对映异构体”或者“旋光异构体”是指互为镜像关系的立体异构体。
除非另有说明，术语“顺反异构体”或者“几何异构体”系由因双键或者成环碳原子单键不能自由旋转而引起。
除非另有说明，术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。
除非另有说明，“(D)”或者“(+)”表示右旋，“(L)”或者“(-)”表示左旋，“(DL)”或者“(±)”表示外消旋。
除非另有说明，用楔形实线键
Figure PCTCN2021129783-appb-000025
和楔形虚线键
Figure PCTCN2021129783-appb-000026
表示一个立体中心的绝对构型，用直形实线键
Figure PCTCN2021129783-appb-000027
和直形虚线键
Figure PCTCN2021129783-appb-000028
表示立体中心的相对构型，用波浪线
Figure PCTCN2021129783-appb-000029
表示楔形实线键
Figure PCTCN2021129783-appb-000030
或楔形虚线键
Figure PCTCN2021129783-appb-000031
或用波浪线
Figure PCTCN2021129783-appb-000032
表示直形实线键
Figure PCTCN2021129783-appb-000033
和直形虚线键
Figure PCTCN2021129783-appb-000034
本发明的化合物可以存在特定的。除非另有说明，术语“互变异构体”或“互变异构体形式”是指在室温下，不同官能团异构体处于动态平衡，并能很快的相互转化。若互变异构体是可能的(如在溶液中)，则可以达到互变异构体的化学平衡。例如，质子互变异构体(proton tautomer)(也称质子转移互变异构体(prototropic tautomer))包括通过质子迁移来进行的互相转化，如酮-烯醇异构化和亚胺-烯胺异构化。价键异构体(valence tautomer)包括一些成键电子的重组来进行的相互转化。其中酮-烯醇互变异构化的具体实例 是戊烷-2,4-二酮与4-羟基戊-3-烯-2-酮两个互变异构体之间的互变。
除非另有说明，术语“富含一种异构体”、“异构体富集”、“富含一种对映体”或者“对映体富集”指其中一种异构体或对映体的含量小于100％，并且，该异构体或对映体的含量大于等于60％，或者大于等于70％，或者大于等于80％，或者大于等于90％，或者大于等于95％，或者大于等于96％，或者大于等于97％，或者大于等于98％，或者大于等于99％，或者大于等于99.5％，或者大于等于99.6％，或者大于等于99.7％，或者大于等于99.8％，或者大于等于99.9％。
除非另有说明，术语“异构体过量”或“对映体过量”指两种异构体或两种对映体相对百分数之间的差值。例如，其中一种异构体或对映体的含量为90％，另一种异构体或对映体的含量为10％，则异构体或对映体过量(ee值)为80％。
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。
发明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。例如，可用放射性同位素标记化合物，比如氚( 3H)，碘-125( 125I)或C-14( 14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。
术语“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。
术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，取代基可以包括重氢和氢的变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧(即＝O)时，意味着两个氢原子被取代。氧取代不会发生在芳香基上。术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。
当一个连接基团的数量为0时，比如-(CRR) 0-，表示该连接基团为单键。
当其中一个变量选自单键时，表示其连接的两个基团直接相连，比如A-L-Z中L代表单键时表示该结构实际上是A-Z。
一个取代基为空缺时，表示该取代基是不存在的，比如A-X中X为空缺时表示该结构实际上是A。 当所列举的取代基中没有指明其通过哪一个原子连接到被取代的基团上时，这种取代基可以通过其任何原子相键合，例如，吡啶基作为取代基可以通过吡啶环上任意一个碳原子连接到被取代的基团上。
当所列举的连接基团没有指明其连接方向，其连接方向是任意的，例如，
Figure PCTCN2021129783-appb-000035
中连接基团L为-M-W-，此时-M-W-既可以按与从左往右的读取顺序相同的方向连接环A和环B构成
Figure PCTCN2021129783-appb-000036
也可以按照与从左往右的读取顺序相反的方向连接环A和环B构成
Figure PCTCN2021129783-appb-000037
所述连接基团、取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。
除非另有规定，当某一基团具有一个或多个可连接位点时，该基团的任意一个或多个位点可以通过化学键与其他基团相连。当该化学键的连接方式是不定位的，且可连接位点存在H原子时，则连接化学键时，该位点的H原子的个数会随所连接化学键的个数而对应减少变成相应价数的基团。所述位点与其他基团连接的化学键可以用直形实线键
Figure PCTCN2021129783-appb-000038
直形虚线键
Figure PCTCN2021129783-appb-000039
或波浪线
Figure PCTCN2021129783-appb-000040
表示。例如-OCH 3中的直形实线键表示通过该基团中的氧原子与其他基团相连；
Figure PCTCN2021129783-appb-000041
中的直形虚线键表示通过该基团中的氮原子的两端与其他基团相连；
Figure PCTCN2021129783-appb-000042
中的波浪线表示通过该苯基基团中的1和2位碳原子与其他基团相连。
Figure PCTCN2021129783-appb-000043
表示该哌啶基上的任意可连接位点可以通过1个化学键与其他基团相连，至少包括
Figure PCTCN2021129783-appb-000044
Figure PCTCN2021129783-appb-000045
这4种连接方式，即使-N-上画出了H原子，但是
Figure PCTCN2021129783-appb-000046
仍包括
Figure PCTCN2021129783-appb-000047
这种连接方式的基团，只是在连接1个化学键时，该位点的的H会对应减少1个变成相应的一价哌啶基。
除非另有规定，术语“C 1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C 1-3烷基包括C 1-2和C 2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-3烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)等。
除非另有规定，术语“C 1-3烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至3个碳原子的烷基基团。所述C 1-3烷氧基包括C 1-2、C 2-3、C 3和C 2烷氧基等。C 1-3烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)等。
除非另有规定，C n-n+m或C n-C n+m包括n至n+m个碳的任何一种具体情况，例如C 1-12包括C 1、C 2、C 3、C 4、C 5、C 6、C 7、C 8、C 9、C 10、C 11、和C 12，也包括n至n+m中的任何一个范围，例如C 1-12包括C 1- 3、C 1-6、C 1-9、C 3-6、C 3-9、C 3-12、C 6-9、C 6-12、和C 9-12等；同理，n元至n+m元表示环上原子数为n至n+m个，例如3-12元环包括3元环、4元环、5元环、6元环、7元环、8元环、9元环、10元环、11元环、和12元环，也包括n至n+m中的任何一个范围，例如3-12元环包括3-6元环、3-9元环、5-6元环、5-7元环、6-7元环、6-8元环、和6-10元环等。
术语“离去基团”是指可以被另一种官能团或原子通过取代反应(例如亲和取代反应)所取代的官能团或原子。例如，代表性的离去基团包括三氟甲磺酸酯；氯、溴、碘；磺酸酯基，如甲磺酸酯、甲苯磺酸酯、对溴苯磺酸酯、对甲苯磺酸酯等；酰氧基，如乙酰氧基、三氟乙酰氧基等等。
术语“保护基”包括但不限于“氨基保护基”、“羟基保护基”或“巯基保护基”。术语“氨基保护基”是指适合用于阻止氨基氮位上副反应的保护基团。代表性的氨基保护基包括但不限于：甲酰基；酰基，例如链烷酰基(如乙酰基、三氯乙酰基或三氟乙酰基)；烷氧基羰基，如叔丁氧基羰基(Boc)；芳基甲氧羰基，如苄氧羰基(Cbz)和9-芴甲氧羰基(Fmoc)；芳基甲基，如苄基(Bn)、三苯甲基(Tr)、1,1-二-(4'-甲氧基苯基)甲基；甲硅烷基，如三甲基甲硅烷基(TMS)，2-(三甲基硅)乙氧基甲基(SEM)和叔丁基二甲基甲硅烷基(TBS)等等。术语“羟基保护基”是指适合用于阻止羟基副反应的保护基。代表性羟基保护基包括但不限于：烷基，如甲基、乙基和叔丁基；酰基，例如链烷酰基(如乙酰基)；芳基甲基，如苄基(Bn)，对甲氧基苄基(PMB)、9-芴基甲基(Fm)和二苯基甲基(二苯甲基，DPM)；甲硅烷基，如三甲基甲硅烷基(TMS)和叔丁基二甲基甲硅烷基(TBS)等等。
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。
本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：
Figure PCTCN2021129783-appb-000048
扫描，收集相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。
本发明所使用的溶剂可经市售获得。
本发明采用下述缩略词：g代表克；mg代表毫克；μL代表微升；mL代表毫升；mol代表摩尔；mmol代表毫摩尔；μmol代表微摩尔；M代表摩尔/升；mM代表毫摩尔/升；μM代表微摩尔/升；nM代表纳摩尔/升。Me代表甲基；Boc代表叔丁氧羰基；DMSO代表二甲亚砜；DMSO-d 6代表氘代二甲亚砜。
本发明化合物依据本领域常规命名原则或者使用
Figure PCTCN2021129783-appb-000049
软件命名，市售化合物采用供应商目录名称。
具体实施方式
下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。
中间体A
Figure PCTCN2021129783-appb-000050
合成路线：
Figure PCTCN2021129783-appb-000051
第一步
将化合物A-1(3.45g,16.4mmol)，化合物A-2(4.76g,24.5mmol)，三(二亚苄基丙酮)二钯(1.50g,1.63mmol)，2-二环己基膦-2’,4’,6’-三异丙基联苯(779mg,1.63mmol)，碳酸钾(6.78g,49.0mmol)溶于二氧六环(80mL)和水(20mL)中，反应液于95摄氏度下搅拌12小时。反应完成后，将反应液直接过滤，滤液减压浓缩，残留物经硅胶柱层析色谱(洗脱剂:石油醚/乙酸乙酯,15/1～1/1,V/V)分离纯化得到化合物A-3。MS-ESI计算值[M+H] +281,实测值281。
第二步
将化合物A-3(3g,9.55mmol)溶于四氢呋喃(32mL)，水(8mL)和甲醇(8mL)中，加入一水合氢氧化锂(1.20g,28.7mmol)，并在28摄氏度下搅拌12小时。反应完成后，将反应液减压浓缩除去四氢呋喃和甲醇，加入水(100mL)进行稀释，用甲基叔丁基醚(60mL×1)萃取。水相用稀盐酸水溶液(1M)调节pH至3，抽滤，滤饼经水洗，干燥得化合物中间体A。MS-ESI计算值[M+H] +253,实测值253。
中间体B
Figure PCTCN2021129783-appb-000052
合成路线：
Figure PCTCN2021129783-appb-000053
第一步
将乌洛托品(30.6g,218mmol)加入到化合物B-1(50.0g,218mmol)的二氯甲烷(625mL)溶液中，反应液在30摄氏度下搅拌1小时。将反应液过滤，收集滤饼得到粗品化合物B-2。
第二步
将化合物B-2(65.0g,176mmol)溶解在乙醇(429mL)中，然后加入35％的浓盐酸(52.0mL)，反应液在30摄氏度下搅拌72小时。反应液过滤，滤液减压浓缩得到化合物B-3的盐酸盐。
第三步
将化合物B-3的盐酸盐(30g,149mmol)溶解在水(375mL)中，然后加入碳酸氢钠(31.2g,372mmol)，甲醇(375mL)和二碳酸二叔丁酯(48.7g,223mmol,51.3mL)。反应液在30摄氏度下搅拌1小时。反应液用乙酸乙酯(600mL x 2)萃取。合并的有机相用无水硫酸钠干燥，过滤，滤液减压浓缩。减压浓缩所得粗品经硅胶柱层析色谱(洗脱剂:石油醚/乙酸乙酯,1/0～5/1,V/V)分离纯化得到化合物B-4。
第四步
将醋酸铵(116g,1.51mol)和氰基硼氢化钠(9.47g,151mmol)加到化合物B-4(40.0g,151mmol)的甲醇(400mL)溶液中。反应液在50摄氏度下搅拌18个小时。将反应液减压浓缩，往剩余物中加入二氯甲烷(600mL)搅拌过滤，滤液减压浓缩。减压浓缩所得粗品使用反相制备色谱柱(氨水体系)纯化得到中间体B。MS-ESI计算值[M+H] +267,实测值267。
中间体C
Figure PCTCN2021129783-appb-000054
合成路线：
Figure PCTCN2021129783-appb-000055
第一步
将化合物C-1(8.00g,40.6mmol)溶于N,N-二甲基乙酰胺(80mL)中，在氮气氛围下向反应液中加入1-氯甲基-4-氟-1,4-二氮杂双环[2.2.2]辛烷二(四氟硼酸)盐(28.8g,81.2mmol)，反应液在60摄氏度下搅拌14小时。将反应液减压浓缩，所得粗品经硅胶柱层析色谱(洗脱剂:石油醚/乙酸乙酯,50/1～25/1,V/V)分离纯化得到化合物C-2。MS-ESI计算值[M+H] +215和217,实测值215和217。
第二步
将化合物C-2(2.30g,10.7mmol)和联硼酸频哪醇酯(5.43g,21.4mmol)溶于二氧六环(50mL)中，加入[1,1′-双(二苯基膦)二茂铁]二氯化钯(II)的二氯甲烷复合物(873mg,1.07mmol)和醋酸钾(2.10g,21.4mmol)，反应液于95摄氏度下搅拌14小时。反应液减压浓缩，所得粗品经柱层析色谱(洗脱剂:石油醚/乙酸乙酯,30/1～10/1,V/V)分离纯化得到中间体C。MS-ESI计算值[M+H] +263,实测值263。
实施例1
Figure PCTCN2021129783-appb-000056
合成路线：
Figure PCTCN2021129783-appb-000057
第一步
将化合物1-1(6.00g,39.7mmol)溶于二氯甲烷(20mL)中，加入N-溴代琥珀酰亚胺(7.06g,39.7mmol)和对甲基苯磺酸(1.37g,7.94mmol),反应液在50摄氏度下搅拌反应12小时。将反应液用水(150mL)稀释，加入乙酸乙酯(250mL ×2)萃取，合并有机相。有机相用饱和食盐水(200mL)洗涤一次，用无水硫酸钠干燥，过滤，滤液减压浓缩。减压浓缩后的粗品经硅胶柱层析色谱(洗脱剂:石油醚/乙酸乙酯,100/1～20/1,V/V)分离纯化得到化合物1-2。
第二步
将乌洛托品(2.68g,19.1mmol)加入到化合物1-2(4.40g,19.1mmol)的二氯甲烷(60mL)溶液中，反应液在20摄氏度下搅拌1小时。将反应液过滤，收集滤饼得到粗品化合物1-3。
第三步
将化合物1-3(3.00g,8.17mmol)溶解在乙醇(30mL)中，然后加入35％的浓盐酸(2.41mL)，反应液在80摄氏度下搅拌2小时。反应液过滤，将滤液减压浓缩干燥得到化合物1-4的盐酸盐。
第四步
将化合物1-4的盐酸盐(2.80g,13.8mmol)溶解在水(25mL)和四氢呋喃(25mL)中，然后加入二碳酸二叔丁酯(2.41g,11.1mmol,2.54mL)和碳酸氢钠(3.58g,41.5mmol)。反应液在20摄氏度下搅拌1小时。反应液加水(60mL)稀释，用乙酸乙酯(100mL x 2)萃取。合并的有机相用饱和食盐水(100mL)洗涤一次，用无水硫酸钠干燥，过滤，滤液减压浓缩。减压浓缩后的粗品经硅胶柱层析色谱(洗脱剂:石油醚/乙酸乙酯,30/1～10/1,V/V)分离纯化得到化合物1-5。
第五步
将醋酸铵(409mg,5.30mol)和氰基硼氢化钠(33.3mg,530mmol)加到化合物1-5(300mg,530mmol)的甲醇(10mL)溶液中。反应液在50摄氏度下搅拌1个小时。将反应液过滤，滤液减压浓缩。剩余物经制备高效液相色谱(色谱柱：Waters Xbridge 150 x 25mm x 5μm；流动相：[10mM的碳酸氢铵水溶液-乙腈]；B％：20％-50％，10分钟)分离纯化得到化合物1-6。MS-ESI计算值[M+H] +268,实测值268。
第六步
将化中间体A(151mg,599μmol)溶于N,N-二甲基甲酰胺(8mL)中，然后加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(296mg,778μmol)，N,N-二异丙基乙胺(232mg,1.80mmol,313μL)和化合物1-6(160mg,599μmol)。反应液在20摄氏度下搅拌12小时。将反应液用水(80mL)稀释，用乙酸乙酯(100mL x 2)萃取，合并有机相。有机相用饱和食盐水(80mL)洗涤一次，用无水硫酸钠干燥，过滤，滤液减压浓缩。剩余物用薄层层析法(展开剂:乙酸乙酯)分离得到化合物1-7。
第七步
将化合物1-7(220mg,420μmol)溶于二氯甲烷(5mL)中，加入三氟乙酸(2mL)，反应液在20摄氏度下搅拌反应1个小时。将反应液减压浓缩，剩余物经制备高效液相色谱(色谱柱：Phenomenex Synergi C18 150 x 30mm x 4μm；流动相：[0.05％的盐酸水溶液-乙腈]；B％：20％-40％，10分钟)分离纯化得到化合物1的盐酸盐。
MS-ESI计算值[M+H] +402,实测值402。 1H NMR(400MHz,DMSO-d 6)δ9.31(d,J＝8.0Hz,1H),8.40(s,1H),8.32(s,3H),8.16(s,1H),7.99-7.88(m,2H),7.78(dd,J＝1.6,8.8Hz,1H),7.72(dd,J＝7.6,8.0Hz,1H),7.63-7.53(m,2H),7.06(d,J＝7.3Hz,1H),6.76(d,J＝8.0Hz,1H),5.48-5.37(m,1H),3.90(s,3H),3.61-3.24(m,2H),2.56(s,3H)ppm。
实施例2
Figure PCTCN2021129783-appb-000058
合成路线：
Figure PCTCN2021129783-appb-000059
第一步
将化合物2-1(1g,7.29mmol)和叔丁基亚磺酰胺(928mg,7.66mmol)溶于二氯甲烷(10mL)中，加入碳酸铯(4.75g,14.6mmol)，反应液在20摄氏度下搅拌反应15个小时。反应液用硅藻土过滤，滤液减压浓缩得粗品化合物2-2。MS-ESI计算值[M+H] +241,实测值241。
第二步
将化合物硝基甲烷(3.55mL,65.8mmol)溶于二甲亚砜(15mL)中，加入叔丁醇钠(160mg,1.66mmol)，反应液在30摄氏度下搅拌反应1个小时，加入化合物2-2(2g,8.32mmol)，反应液在30摄氏度下继续搅拌反应15个小时。反应液加入饱和氯化铵水溶液(100mL)，用乙酸乙酯(30mL x 4)萃取，合并有机相。有机相用无水硫酸钠干燥，过滤，滤液减压浓缩得粗品化合物2-3。MS-ESI计算值[M+H] +302,实测值302。
第四步
将化合物2-3(2g,6.64mmol)溶于乙酸乙酯(3mL)中，加入4M的氯化氢乙酸乙酯溶液(13.3mL)，反应液在15摄氏度下搅拌反应16个小时。将反应液减压浓缩，浓缩所得粗品加入乙酸乙酯(50mL)，搅拌0.5个小时，过滤得到化合物2-4的盐酸盐。
第五步
将化合物2-4的盐酸盐(250mg,1.07mmol)，中间体A(270mg,1.07mmol)，1-乙基-(3-二甲基氨基丙基)碳二亚胺盐酸盐(410mg,2.14mmol)和1-羟基苯并三唑(217mg,1.60mmol)溶于四氢呋喃(4mL)和N,N-二甲基甲酰胺(1mL)中，向反应液中添加N,N-二异丙基乙胺(553mg,4.28mmol,745μL)。反应液在30摄氏度下搅拌1小时。将反应液减压浓缩，所得粗品用薄层层析法(展开剂:二氯甲烷/甲醇,10/1,V/V))分离得到化合物2-5。MS-ESI计算值[M+H] +432,实测值432。
第六步
将化合物2-5(120mg,220μmol)溶于乙醇(2mL)和水(0.5mL)中，加入铁粉(77.7mg,1.39mmol)和氯化铵(149mg,2.78mmol)，反应液在95摄氏度下搅拌反应2个小时。反应液加压浓缩，剩余物加入二氯甲烷/甲醇(10/1,V/V)溶液(50mL)，过滤，滤液减压浓缩。减压浓缩得到的粗品，经过制备高效液相色谱(色谱柱：Xtimate C18 150x 25mm x 5μm；流动相：[0.05％的碳酸氢铵水溶液-乙腈]；B％：21％-41％，10分钟)分离纯化得到化合物2。
MS-ESI计算值[M+H] +402,实测值402。 1H NMR(400MHz,DMSO-d 6)δ8.36(d,J＝6.0Hz,1H),8.24(s,1H),8.05(s,1H),7.93-7.81(m,2H),7.74(dd,J＝1.6,8.8Hz,1H),7.61-7.54(m,2H),7.04-6.94(m,1H),6.87(dd,J＝2.4,5.6Hz,1H),5.14-4.93(m,1H),3.82(s,3H),3.17-2.92(m,2H),2.56(s,3H)ppm。
实施例3
Figure PCTCN2021129783-appb-000060
合成路线：
Figure PCTCN2021129783-appb-000061
第一步
参考实施例2第一步得到化合物3-2。MS-ESI计算值[M+H] +211,实测值211。
第二步
参考实施例2第二步得到化合物3-3。
第三步
参考实施例2第三步得到化合物3-4的盐酸盐。
第四步
参考实施例2第四步得到化合物3-5。MS-ESI计算值[M+H] +402,实测值402。
第五步
参考实施例2第五步得到粗品，所得粗品经过制备高效液相色谱(色谱柱：Phenomenex Synergi C18 150x 30mm x 4μm；流动相：[0.05％的盐酸水溶液-乙腈]；B％：7％-27％，11分钟)分离纯化得到化合物3的盐酸盐。
MS-ESI计算值[M+H] +372,实测值372。 1H NMR(400MHz,DMSO-d 6)δ9.92(d,J＝7.6Hz,1H),9.17(s,1H),8.98-8.76(m,2H),8.60(s,3H),8.50(s,1H),8.28(s,1H),8.18-8.05(m,1H),7.93(t,J＝8.4Hz,2H),7.83(d,J＝8.8Hz,1H),7.66-7.47(m,2H),5.76-5.59(m,1H),3.74-3.39(m,2H),2.58(s,3H)ppm。
实施例4
Figure PCTCN2021129783-appb-000062
合成路线：
Figure PCTCN2021129783-appb-000063
第一步
参考实施例2第一步得到化合物4-2。MS-ESI计算值[M+H] +241,实测值241。
第二步
参考实施例2第二步得到化合物4-3。MS-ESI计算值[M+H] +302,实测值302。
第三步
参考实施例2第三步得到化合物4-4的盐酸盐。
第四步
参考实施例2第四步得到化合物4-5。MS-ESI计算值[M+H] +432,实测值432。
第五步
参考实施例2第五步得到化合物4。
MS-ESI计算值[M+H] +402,实测值402。 1H NMR(400MHz,DMSO-d 6)δ12.71(s,1H),8.85(d,J＝8.0Hz,1H),8.28-8.15(m,3H),8.04(s,1H),7.93-7.81(m,2H),7.72(dd,J＝1.6,8.4Hz,1H),7.61-7.49(m,2H),7.45-7.38(m,1H),5.08-4.91(m,1H),3.83(s,3H),3.09-2.81(m,2H),2.55(s,3H)ppm。
实施例5
Figure PCTCN2021129783-appb-000064
合成路线：
Figure PCTCN2021129783-appb-000065
第一步
参考实施例1第六步得到化合物5-2。MS-ESI计算值[M+H] +451,453,实测值451,453。
第二步
将化合物5-2(60.0mg,133μmol)和，化合物5-3(41.2mg,160μmol)碳酸钾(36.8mg,266μmol)加入二氧六环(4.5mL)和水(0.5mL)中，然后再加入[1,1′-双(二苯基膦)二茂铁]二氯化钯(II)(9.73mg,13.3μmol)，反应液于90摄氏度下搅拌16小时。反应液减压浓缩得粗品，粗品经硅胶柱层析色谱(洗脱剂:二氯甲烷/甲醇,200/1～10/1,V/V)分离纯化得到化合物5-4。MS-ESI计算值[M+H] +503,实测值503。
第三步
参考实施例1第七步得到化合物5的盐酸盐。
MS-ESI计算值[M+H] +403,实测值403。 1H NMR(400MHz,DMSO-d 6)δ9.56(d,J＝8.8Hz,1H),8.75(d,J＝5.2Hz,1H),8.56-8.42(m,1H),8.38-8.27(m,1H),8.21-8.03(m,3H),7.85(d,J＝8.4Hz,1H),7.74(t,J＝7.6Hz,1H),7.62(d,J＝8.8Hz,1H),7.06(d,J＝7.6Hz,1H),6.80(d,J＝8.0Hz,1H),5.54-5.40(m,1H),3.92(s,3H),3.58-3.42(m,2H),2.64-2.54(m,3H)ppm.。
实施例6
Figure PCTCN2021129783-appb-000066
合成路线：
Figure PCTCN2021129783-appb-000067
第一步
将化合物6-1(1g,3.80mmol)溶于N,N-二甲基甲酰胺(8mL)中，加入1-乙基-(3-二甲基氨基丙基)碳二亚胺盐酸盐(1.12g,5.85mmol)，1-羟基苯并三唑(579mg,4.29mmol)和氨水(1.82g,13.0mmol,2.00mL,25％纯度)。反应液在20摄氏度下搅拌15小时。往反应液中加入水(60mL)，反应液在0摄氏度下搅拌1个小时，过滤，收集滤饼得到粗品化合物6-2。
第二步
将化合物6-2(470mg,1.79mmol)加入三甲氧基甲烷(4.84g,45.6mmol,5mL)和N-甲基吡咯烷酮(1mL)中，再加入对甲基苯磺酸(34.1mg,179μmol)。反应液在110摄氏度下搅拌反应12个小时。往反应液中加入水(60mL)，反应液继续在0摄氏度下搅拌1个小时。反应液过滤，收集滤饼得到粗品化合物6-3。
第三步
将化合物6-3(370mg,1.36mmol)加入N,N-二甲基甲酰胺(0.1mL)中，再加入氯化亚砜(4mL)，反应液在90摄氏度下搅拌反应14个小时。将反应液冷却至室温后减压浓缩。往减压浓缩所得剩余物中加入甲苯(20mL)，用饱和碳酸氢钠(20mL)洗涤一次。有机相用无水硫酸钠干燥，过滤，滤液减压浓缩得到粗品化合物6-4。
MS-ESI计算值[M+H] +291,实测值291。
第四步
将化合物6-4(220mg,757μmol)和中间体B(205mg,757μmol)溶于异丙醇(5mL)中，加入碳酸钠(161mg,1.51mmol)。反应液在60摄氏度下搅拌反应15个小时。将反应液减压浓缩，浓缩所得粗品经柱层析色谱(洗脱剂:石油醚/乙酸乙酯,10/1～4/1,V/V)分离纯化得到化合物6-5。
MS-ESI计算值[M+H] +521,实测值521。
第五步
将化合物6-5(200mg,384μmol)，中间体C(101mg,384μmol)，磷酸钾(163mg,769μmol)和[1,1′-双(二苯基膦)二茂铁]二氯化钯(II)的二氯甲烷复合物(62.8mg,76.9μmol)加入到N,N-二甲基甲酰胺(3mL)和水(0.3mL)中。反应液在85摄氏度下搅拌反应16个小时。将反应液减压浓缩，浓缩所得粗品经柱层析色谱(洗脱剂:石油醚/乙酸乙酯,10/1～1/1,V/V)分离纯化得到化合物6-6。
MS-ESI计算值[M+H] +529,实测值529。
第六步
将化合物6-6(75mg,142μmol)加入到4M的氯化氢乙酸乙酯溶液(5mL)中，反应液在20摄氏度下搅拌反应40分钟。将反应液减压浓缩，浓缩所得粗品经制备高效液相色谱(色谱柱：Phenomenex Synergi C18150x 25mm 10μm；流动相：[0.05％的盐酸水溶液-乙腈]；B％：8％-28％，10分钟)分离纯化得到化合物6的盐酸盐。
MS-ESI计算值[M+H] +429,实测值429。 1H NMR(400MHz,DMSO-d 6)δ13.04-12.67(m,1H),9.42-9.06(m,1H),8.68(s,1H),8.59-8.44(m,3H),8.01(d,J＝7.2Hz,1H),7.89-7.80(m,2H),7.67(d,J＝8.8Hz,1H),7.58-7.49(m,1H),7.38-7.23(m,2H),7.19-7.09(m,1H),6.96-6.85(m,1H),5.98(d,J＝6.4Hz,1H),4.09-3.82(m,1H),3.79-3.70(m,3H),3.45-3.21(m,1H)ppm。
实施例7
Figure PCTCN2021129783-appb-000068
合成路线：
Figure PCTCN2021129783-appb-000069
第一步
参考实施例6第五步得到化合物7-1。
MS-ESI计算值[M+H] +525,实测值525。
第二步
参考实施例6第六步得到化合物7的盐酸盐。
MS-ESI计算值[M+H] +425,实测值425。 1H NMR(400MHz,DMSO-d 6)δ11.42(d,J＝7.6Hz,1H),9.29(d,J＝8.4Hz,1H),8.69(s,1H),8.58(s,3H),8.02(dd,J＝1.0,7.6Hz,1H),7.92-7.82(m,2H),7.65(d,J＝8.4Hz,1H),7.42(dd,J＝1.6,8.4Hz,1H),7.34-7.27(m,2H),7.18(d,J＝7.8Hz,1H),6.90(dd,J＝2.0,8.0Hz,1H),6.10-5.93(m,1H),4.10-3.87(m,1H),3.76(s,3H),3.35(dd,J＝4.0,7.2Hz,1H),2.52(s,3H)ppm。
实施例8
Figure PCTCN2021129783-appb-000070
合成路线：
Figure PCTCN2021129783-appb-000071
第一步
将化合物8-1(4.4g,17.3mmol)溶于二氯甲烷(40mL)中，反应液冷却至0摄氏度，分批加入间氯过氧苯甲酸(5.25g,25.9mmol,85％纯度)。反应液在20摄氏度下搅拌反应15个小时。将反应液用二氯甲烷(100mL)稀释，依次用10％碳酸氢钠水溶液(100mL)、10％的硫代硫酸钠水溶液(100mL)和饱和食盐水(100mL) 洗涤。有机相用无水硫酸钠干燥，过滤，滤液减压浓缩。浓缩所得粗品经硅胶柱层析色谱(洗脱剂:二氯甲烷/甲醇,100/1～10/1,V/V)分离纯化得到化合物8-2。
MS-ESI计算值[M+H] +272,实测值272。
第二步
将化合物8-2(1g,3.69mmol)溶于二氯甲烷(20mL)中，加入三乙胺(747mg,7.38mmol,1.03mL)，将反应液冷却至零下70摄氏度，缓慢滴加草酰氯(937mg,7.38mmol,646μL)。反应液在零下70摄氏度搅拌反应0.5个小时，升温至20摄氏度，继续搅拌反应2个小时。往反应液中加入甲醇(5mL)稀释，所得有机相依次用饱和碳酸氢钠(50mL)和水(50mL)洗涤。有机相用无水硫酸钠干燥，过滤，滤液减压浓缩。浓缩所得粗品经柱层析色谱(洗脱剂:石油醚/乙酸乙酯,100/1～50/1,V/V)分离纯化得到化合物8-3。
MS-ESI计算值[M+H] +290,实测值290。
第三步
参考实施例6第五步得到化合物8-4。
MS-ESI计算值[M+H] +298,实测值298。
第四步
参考实施例6第四步得到化合物8-5。
MS-ESI计算值[M+H] +528,实测值528。
第五步
参考实施例6第六步得到化合物8的盐酸盐。
MS-ESI计算值[M+H] +428,实测值428。 1H NMR(400MHz,DMSO-d 6)δ10.33(s,1H),9.31(d,J＝8.4Hz,1H),8.55(s,3H),8.03-7.96(m,1H),7.95-7.88(m,1H),7.77(s,1H),7.62(dd,J＝3.6,7.6Hz,2H),7.45-7.26(m,4H),7.08(d,J＝7.2Hz,1H),6.92(d,J＝7.6Hz,1H),5.85-5.75(m,1H),4.02-3.85(m,1H),3.77(s,3H),3.47-3.35(m,1H)ppm。
实施例9
Figure PCTCN2021129783-appb-000072
合成路线：
Figure PCTCN2021129783-appb-000073
第一步
参考实施例6第五步得到化合物9-1。
MS-ESI计算值[M+H] +294,实测值294。
第二步
参考实施例6第四步得到化合物9-2。
MS-ESI计算值[M+H] +524,实测值524。
第三步
参考实施例6第六步得到化合物9的盐酸盐。
MS-ESI计算值[M+H] +424,实测值424。 1H NMR(400MHz,DMSO-d 6)δ12.81(d,J＝7.6Hz,1H),9.29-9.02(m,1H),8.76-8.29(m,3H),8.01-7.84(m,2H),7.76(s,1H),7.71-7.61(m,2H),7.55-7.44(m,1H),7.39-7.20(m,3H),7.02(d,J＝5.6Hz,1H),6.94-6.88(m,1H),5.84-5.70(m,1H),3.99-3.71(m,4H),3.50-3.35(m,1H)ppm。
活性测试
1.体外评价ROCK蛋白激酶抑制活性
实验目的：检测化合物的ROCK蛋白激酶抑制IC 50值。
实验材料:
测定緩沖溶液:20mM 4-羟乙基哌嗪乙磺酸(pH 7.5)，10mM氯化镁，1mM乙二醇二乙醚二胺四乙酸，0.02％聚氧乙烯月桂醚，0.02mg/mL牛血清白蛋白，0.1mM钒酸钠，2mM二硫苏糖醇，1％DMSO。实验操作:
将新鲜制备的缓冲溶液中加入ROCK2蛋白激酶底物Long S6 Kinase substrate peptide，浓度20μM，然后加入1nM ROCK2蛋白激酶，均匀搅拌。使用Echo550加入含有待测化合物的系列DMSO稀释液(始于10μM，按3倍系列稀释)，室温下预温育20分钟，加入 33P-ATP(放射强度10μCi/μL)引发反应，室温反应两小时。然后使用P81离子交换纸(Whatman#3698-915)过滤，用0.75％磷酸洗涤。使用Filter-Binding方法检测放射强度。
化合物的蛋白激酶抑制活性表达为相对空白底物(单纯DMSO)残存的蛋白激酶活性。利用Prism软件包(GraphPad Software,San Diego California,USA)计算IC 50值和曲线。
实验结果：
表1 ROCK抑制活性测试结果
化合物 对ROCK2的IC 50(nM)
1(盐酸盐) 8
3(盐酸盐) 45
4 54
5(盐酸盐) 27
6(盐酸盐) 56
7(盐酸盐) 50
8(盐酸盐) 63
9(盐酸盐) 72
本发明化合物具有显著甚至意料不到的ROCK2抑制活性。

Claims (13)

  1. 式(I)所示化合物或其药学上可接受的盐，
    Figure PCTCN2021129783-appb-100001
    其中，
    T 1为N或CH；T 3为N或CR 3；
    当T 2为O时，T 3为N，结构单元
    Figure PCTCN2021129783-appb-100002
    为
    Figure PCTCN2021129783-appb-100003
    当T 2为N时，结构单元
    Figure PCTCN2021129783-appb-100004
    为
    Figure PCTCN2021129783-appb-100005
    T 4为N或CH；
    R 1、R 2、R 3和R 4各自独立地为H、F、Cl、Br、I、-OH、-CN、-NH 2、-NO 2、C 1-3烷基或C 1-3烷氧基，其中所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN、-NH 2或-NO 2的取代基所取代；
    R 6和R 7各自独立地为H、F、Cl或C 1-3烷基，其中所述C 1-3烷基任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN、-NH 2或-NO 2的取代基所取代；
    各R 8独立地为H、F、Cl、Br、I、-OH、-CN、-NH 2、-NO 2、C 1-3烷基或C 1-3烷氧基，其中所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN、-NH 2或-NO 2的取代基所取代；n为1、2、3或4。
  2. 根据权利要求1所述的化合物或其药学上可接受的盐，其化合物具有式(I-1)或(I-2)所示结构：
    Figure PCTCN2021129783-appb-100006
    其中，
    R 1、R 2、R 3、R 4、R 6、R 7、R 8、T 1、T 3、T 4和n如权利要求1所定义。
  3. 根据权利要求1或2所述的化合物或其药学上可接受的盐，其中R 1、R 2、R 3和R 4各自独立地为H、F、Cl、Br和-CH 3。
  4. 根据权利要求1或2所述的化合物或其药学上可接受的盐，其中R 6和R 7各自独立地为H和-CH 3，其中所述-CH 3任选被1、2或3个独立选自F、Cl、Br、-OH或-NH 2的取代基所取代。
  5. 根据权利要求4所述的化合物或其药学上可接受的盐，其中R 6和R 7各自独立地为H和-CH 2NH 2。
  6. 根据权利要求1或2所述的化合物或其药学上可接受的盐，其中各R 8独立地为H、F、Cl、Br、-OH、-CN、-NH 2、-NO 2或-OCH 3。
  7. 根据权利要求1或2所述的化合物或其药学上可接受的盐，其中结构单元
    Figure PCTCN2021129783-appb-100007
    为
    Figure PCTCN2021129783-appb-100008
  8. 根据权利要求7所述的化合物或其药学上可接受的盐，其中结构单元
    Figure PCTCN2021129783-appb-100009
    为
    Figure PCTCN2021129783-appb-100010
    Figure PCTCN2021129783-appb-100011
  9. 根据权利要求8所述的化合物或其药学上可接受的盐，其中结构单元
    Figure PCTCN2021129783-appb-100012
    为
    Figure PCTCN2021129783-appb-100013
    Figure PCTCN2021129783-appb-100014
  10. 根据权利要求1～7任一项所述的化合物或其药学上可接受的盐，其化合物具有式(I-3)～(I-7)所示结构：
    Figure PCTCN2021129783-appb-100015
    Figure PCTCN2021129783-appb-100016
    其中，R 1、R 2、R 3和R 4如权利要求1、2或3所定义；
    R 6和R 7如权利要求1、2、4或5所定义；
    R 8如权利要求1、2或6所定义；
    n如权利要求1所定义。
  11. 根据权利要求10所述的化合物或其药学上可接受的盐，其化合物具有式(I-8)～(I-12)所示结构：
    Figure PCTCN2021129783-appb-100017
    其中，R 1、R 6、R 7、R 8和n如权利要求10所定义。
  12. 根据权利要求11所述的化合物或其药学上可接受的盐，其化合物具有式(I-13)～(I-17)所示结构：
    Figure PCTCN2021129783-appb-100018
    Figure PCTCN2021129783-appb-100019
    其中，R 1、R 8和n如权利要求11所定义；带“*”的碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在。
  13. 下式化合物或其药学上可接受的盐，其中化合物选自：
    Figure PCTCN2021129783-appb-100020
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