Benzourea ring derivatives, their preparation methods and applications

Abstract

The benzourea ring derivative represented by formula (I) or a pharmaceutically acceptable salt thereof, and a preparation method and application thereof.

Description

[0001] This application claims the following priority rights:

[0002] CN202011259175.6, November 11, 2020. Technical Field

[0003] The present invention relates to the field of drugs, and particularly to a class of benzourea ring derivatives, their preparation methods and applications, and particularly to the compounds shown in formula (I) and their pharmaceutically acceptable salts. Background Art

[0004] Rho-associated protein kinase (Rho associated kinase, abbreviated as ROCK), belonging to serine / threonine protein kinases, is a downstream target effector molecule of Rho and is widely expressed in the human body. Rho-associated protein kinase (ROCK) is involved in the regulation of myosin light chain (MLC) and is applicable to the treatment of vasodilation. New research supports that ROCK kinase is involved in the regulation of the immune response of TH17 cells and the activation of fibroblasts, and the indication can be extended to the treatment of pulmonary diseases such as pulmonary fibrosis and asthma. Further indication expansion includes autoimmune diseases. The ROCK kinase family includes two subtypes, ROCK1 and ROCK2. ROCK2 kinase is related to inflammatory and fibrotic effects. Selective ROCK2 inhibitors do not cause vasodilation even at high concentrations in in vitro vasodilation experiments, and can reduce cardiovascular side effects. Although the embryonic mortality rate of ROCK1 knockout mice is not high, most of them die after birth due to cytoskeletal variation caused by reduced MLC phosphorylation. However, 90% of ROCK2 knockout mice die during the embryonic period, but the surviving mice have no difference from the wild type. Selective inhibition of the activity of ROCK2 may have higher safety. Therefore, selective ROCK2 protein kinase inhibitors can avoid the cardiovascular side effects of drugs.

[0005] KD025 (WO2006105081; WO2008054599; WO2010104851; WO2014055996) is an orally active and selective inhibitor of ROCK2 kinase developed by Kadmon. Research has shown that KD025 represents a new mechanism for treating idiopathic pulmonary fibrosis (IPF) by inhibiting proteins such as RHO kinases that regulate fibrosis. The cause of idiopathic pulmonary fibrosis (IPF) may be tissue damage. The body's response to injury involves reorganization of the actin cytoskeleton in a variety of cells (such as epithelial cells, fibroblasts, endothelial cells, and macrophages), and the assembly of actin filaments and the contraction of actomyosin are regulated by proteins of the RHO kinase family (including ROCK1 and ROCK2). Previous studies have shown that proteins of the RHO kinase family are activated in the lungs of IPF patients and animal models of this disease, and RHO kinase inhibitors can prevent the process of tissue fibrosis in these models and can induce regression of established fibrosis. Currently, a Phase II clinical trial for the treatment of moderate to severe psoriasis has been completed, and it is in the Phase II clinical research stage for the treatment of idiopathic pulmonary fibrosis (IPF).

[0006]

[0007] WO2014134388 and WO2016028971 also disclose a class of compounds with the general structural formulas shown in Formulas (a) and (b), and such compounds can also be used for the treatment of cardiovascular diseases, neuropathological diseases, tumors, autoimmune diseases, pulmonary fibrosis, inflammatory diseases, etc.

[0008]

[0009] The compounds of the present invention are ROCK2 inhibitors and have significant kinase inhibitory activity. SUMMARY OF THE INVENTION

[0010] On the one hand, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0011]

[0012] wherein,

[0013] T1 is N or CH; T3 is N or CR3;

[0014] When T2 is O, T3 is N, and the structural unit is

[0015] When T2 is N, the structural unit is

[0016] T4 is N or CH;

[0017] R1, R2, R3 and R4 are each independently H, F, Cl, Br, I, -OH, -CN, -NH2, -NO2, C 1-3 alkyl or C 1-3 alkoxy, wherein said C 1-3 alkyl and C 1-3 alkoxy are optionally substituted by 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH, -OCH3, -CN, -NH2 or -NO2;

[0018] R6 and R7 are each independently H, F, Cl or C 1-3 alkyl, wherein said C 1-3 alkyl is optionally substituted by 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH, -OCH3, -CN, -NH2 or -NO2;

[0019] Each R8 is independently H, F, Cl, Br, I, -OH, -CN, -NH2, -NO2, C 1-3 alkyl or C 1-3 alkoxy, wherein said C 1-3 alkyl and C 1-3 alkoxy are optionally substituted by 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH, -OCH3, -CN, -NH2 or -NO2; n is 1, 2, 3 or 4.

[0020] In some embodiments of the present invention, the above compound or its pharmaceutically acceptable salt has the structure shown in formula (I-1) or (I-2):

[0021]

[0022] wherein, R1, R2, R3, R4, R6, R7, R8, T1, T3, T4 and n are as defined in the present invention.

[0023] In some embodiments of the present invention, the above R1, R2, R3 and R4 are each independently H, F, Cl, Br and -CH3, and other variables are as defined in the present invention.

[0024] In some embodiments of the present invention, the above R6 and R7 are each independently H and -CH3, wherein said -CH3 is optionally substituted by 1, 2 or 3 substituents independently selected from F, Cl, Br, -OH or -NH2, and other variables are as defined in the present invention.

[0025] In some embodiments of the present invention, R6 and R7 are each independently H and -CH2NH2, and other variables are as defined in the present invention.

[0026] In some embodiments of the present invention, R8 is independently H, F, Cl, Br, -OH, -CN, -NH2, -NO2 or -OCH3, and other variables are as defined in the present invention.

[0027] In some embodiments of the present invention, the above structural unit is n, R8 and other variables are as defined in the present invention.

[0028] In some embodiments of the present invention, the above structural unit is R8 and other variables are as defined in the present invention.

[0029] In some embodiments of the present invention, the above structural unit is Other variables are as defined in the present invention.

[0030] In some embodiments of the present invention, the above compound or a pharmaceutically acceptable salt thereof has a structure represented by formula (I-3) to (I-7):

[0031]

[0032]

[0033] Wherein, R1, R2, R3, R4, R6, R7, R8 and n are as defined in the present invention.

[0034] In some embodiments of the present invention, the above compound or a pharmaceutically acceptable salt thereof has a structure represented by formula (I-8) to (I-12):

[0035]

[0036] Wherein, R1, R6, R7, R8 and n are as defined in the present invention.

[0037] In some embodiments of the present invention, the above compound or a pharmaceutically acceptable salt thereof has a structure represented by formula (I-13) to (I-17):

[0038]

[0039]

[0040] Wherein, R1, R8 and n are as defined in the present invention; the carbon atom with an asterisk (*) is a chiral carbon atom, existing in the form of (R) or (S) single enantiomer or rich in one enantiomer form.

[0041] Some embodiments of the present invention are arbitrarily combined by the above variables.

[0042] In some embodiments of the present invention, the above compound or its pharmaceutically acceptable salt, wherein the compound is selected from:

[0043] Technical effects

[0044] The compounds of the present invention have high kinase inhibitory activity against ROCK2.

[0045] Definitions and explanations

[0046] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered indeterminate or unclear without a special definition, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0047] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are within the scope of sound medical judgment, suitable for use in contact with human and animal tissues, without excessive toxicity, irritation, allergic reaction or other problems or complications, and commensurate with a reasonable benefit / risk ratio.

[0048] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, which are prepared from compounds with specific substituents discovered in the present invention and relatively non-toxic acids or bases. When a compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When a compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, where the inorganic acids include, 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, where the organic acids include, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid and similar 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 can thus be converted into either base or acid addition salts.

[0049] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or basic groups by conventional chemical methods. Generally, the preparation method of such salts is to react these compounds in the form of free acids or bases with a stoichiometric amount of appropriate bases or acids in water or organic solvents or a mixture of both.

[0050] The compounds of the present invention can 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, diastereoisomers, (D)-isomers, (L)-isomers, and their racemic mixtures and other mixtures, such as enantiomer- or diastereomer-enriched mixtures, all of which mixtures are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All these isomers and their mixtures are included within the scope of the present invention.

[0051] Unless otherwise specified, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of each other.

[0052] Unless otherwise specified, the terms "cis-trans isomers" or "geometric isomers" are caused by the inability of double bonds or single bonds of ring carbon atoms to rotate freely.

[0053] Unless otherwise specified, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and are non-mirror images of each other.

[0054] Unless otherwise specified, "(D)" or "(+)" indicates dextrorotation, "(L)" or "(-)" indicates levorotation, and "(DL)" or "(±)" indicates racemization.

[0055] Unless otherwise specified, a solid wedge bond and a dashed wedge bond are used to represent the absolute configuration of a stereocenter, and a solid straight bond and a dashed straight bond are used to represent the relative configuration of a stereocenter. A wavy line represents a solid wedge bond or a dashed wedge bond or a wavy line represents a solid straight bond and a dashed straight bond

[0056] The compounds of the present invention may exist in specific forms. Unless otherwise specified, the term "tautomer" or "tautomeric form" refers to different functional group isomers that are in dynamic equilibrium at room temperature and can rapidly interconvert. If tautomers are possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also known as prototropic tautomers) include interconversions that occur through proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions that occur through the reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between the two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0057] Unless otherwise specified, the terms "enriched in one isomer", "isomer enrichment", "enriched in one enantiomer", or "enantiomer enrichment" mean that the content of one isomer or enantiomer is less than 100%, and the content of this isomer or enantiomer 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%.

[0058] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or 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%, then the isomer or enantiomer excess (ee value) is 80%.

[0059] The optically active (R)- and (S)-isomers as well as D and L 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 the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), it forms a diastereomeric salt with an appropriate optically active acid or base, and then the diastereomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which employs a chiral stationary phase and optionally in combination with chemical derivatization methods (such as generating carbamates from amines).

[0060] The compounds of the invention may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound. For example, the compound can be labeled with a radioactive isotope, such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C). Also, for example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with the non-deuterated drug, the deuterated drug has advantages such as reducing toxic and side effects, increasing drug stability, enhancing efficacy, and prolonging the biological half-life of the drug. 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.

[0061] The term "optionally" or "optionally" means that the subsequently described event or condition may but does not necessarily occur, and this description includes the cases where the described event or condition occurs and the cases where the described event or condition does not occur.

[0062] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by substituents, which may include deuterium and variants of hydrogen, provided that the valence state of the particular atom is normal and the resulting compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it can be substituted or not substituted. Unless otherwise specified, the type and number of substituents can be arbitrary based on what is chemically achievable.

[0063] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted with 0 - 2 Rs, the group may optionally be substituted with up to two Rs, and each R has independent options in each case. In addition, combinations of substituents and / or their variants are only permitted if such combinations result in stable compounds.

[0064] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0065] When one of the variables is selected from a single bond, it means that the two groups it connects are directly linked. For example, when L represents a single bond in A - L - Z, it means that the structure is actually A - Z.

[0066] When a substituent is vacant, it means that the substituent is absent. For example, when X is vacant in A - X, it means that the structure is actually A. When it is not specified which atom of the listed substituent is connected to the group being substituted, such a substituent can be bonded through any of its atoms. For example, a pyridyl group as a substituent can be connected to the group being substituted through any carbon atom on the pyridine ring.

[0067] When the listed linking group does not specify its connection direction, the connection direction is arbitrary. For example, in which the linking group L is -M - W -, at this time -M - W - can connect ring A and ring B in the same direction as the reading order from left to right to form or connect ring A and ring B in the opposite direction to the reading order from left to right to form The combinations of the said linking groups, substituents and / or their variants are only permitted if such combinations result in stable compounds.

[0068] Unless otherwise specified, when a group has one or more connectable sites, any one or more of these sites of the group can be connected to other groups through chemical bonds. When the connection mode of the chemical bond is non - specific and there are H atoms at the connectable sites, then when connecting the chemical bonds, the number of H atoms at this site will correspondingly decrease according to the number of connected chemical bonds to form groups with corresponding valences. The chemical bonds connecting the said site to other groups can be represented by a straight solid line bond a straight dashed line bond or a wavy line For example, the straight solid line bond in -OCH3 represents connection to other groups through the oxygen atom in this group; the straight dashed line bond in The wavy line in [reference] indicates connection to other groups through the 1- and 2-carbon atoms in this phenyl group. It indicates that any connectable site on this piperidinyl group can be connected to other groups through one chemical bond, including at least these 4 connection modes. Even if a hydrogen atom is drawn on the -N-, however it still includes groups with this connection mode. It's just that when connecting one chemical bond, the hydrogen at this site will correspondingly decrease by one to become the corresponding monovalent piperidinyl group.

[0069] Unless otherwise specified, the term "C 1-3 alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 alkyl includes C 1-2 and C 2-3 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene), or polyvalent (such as methine). Examples of C 1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

[0070] Unless otherwise specified, the term "C 1-3 alkoxy" denotes those alkyl groups containing 1 to 3 carbon atoms that are connected to the rest of the molecule through an oxygen atom. The C 1-3 alkoxy includes C 1-2 , C 2-3 , C3, and C2 alkoxy, etc. Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

[0071] Unless otherwise specified, C n-n+m or C n -C n+m includes any specific case of n to n + m carbons. For example, C 1-12 includes C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 , and also includes any range within n to n + m. For example, 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 9-12etc.; similarly, n- to n+m-membered rings mean rings having n to n+m atoms in the ring. For example, 3- to 12-membered rings include 3-membered rings, 4-membered rings, 5-membered rings, 6-membered rings, 7-membered rings, 8-membered rings, 9-membered rings, 10-membered rings, 11-membered rings, and 12-membered rings, and also include any range within n to n+m. For example, 3- to 12-membered rings include 3- to 6-membered rings, 3- to 9-membered rings, 5- to 6-membered rings, 5- to 7-membered rings, 6- to 7-membered rings, 6- to 8-membered rings, and 6- to 10-membered rings, etc.

[0072] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (such as a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chlorine, bromine, iodine; sulfonate groups, such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate, etc.; acyloxy groups, such as acetoxy, trifluoroacetoxy, and so on.

[0073] The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group", or "mercapto protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the nitrogen position of the amino group. Representative amino protecting groups include, but are not limited to: formyl; acyl groups, such as alkanoyl groups (such as acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl groups, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl groups, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl groups, such as benzyl (Bn), trityl (Tr), 1,1-di-(4'-methoxyphenyl)methyl; silyl groups, such as trimethylsilyl (TMS), 2-(trimethylsilyl)ethoxymethyl (SEM), and tert-butyldimethylsilyl (TBS), and so on. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions of the hydroxy group. Representative hydroxy protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanoyl groups (such as acetyl); arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (diphenylmethyl, DPM); silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and so on.

[0074] 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 listed below, the embodiments formed by their combination with other chemical synthesis methods, and equivalent replacement methods well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of the present invention.

[0075] The structure of the compounds 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 a compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD), where the grown single crystal is used to collect diffraction intensity data with a Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode: Scanning. After collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97), and the absolute configuration can be confirmed.

[0076] The solvents used in the present invention are commercially available.

[0077] The following abbreviations are used in the present invention: g represents gram; mg represents milligram; μL represents microliter; mL represents milliliter; mol represents mole; mmol represents millimole; μmol represents micromole; M represents mole per liter; mM represents millimole per liter; μM represents micromole per liter; nM represents nanomole per liter. Me represents methyl; Boc represents tert-butoxycarbonyl; DMSO represents dimethyl sulfoxide; DMSO-d6 represents deuterated dimethyl sulfoxide.

[0078] The compounds of the present invention are named according to the conventional naming principles in the art or using software, and commercially available compounds use the supplier catalog names. Detailed Description of the Invention

[0079] The present invention will be described in detail below by way of examples, but this does not mean any adverse limitation to the present invention. The present invention has been described in detail herein, and specific embodiments thereof have also been disclosed. It will be obvious to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0080] Intermediate A

[0081]

[0082] Synthesis Route:

[0083]

[0084] The First Step

[0085] Compound A-1 (3.45 g, 16.4 mmol), compound A-2 (4.76 g, 24.5 mmol), tris(dibenzylideneacetone)dipalladium(0) (1.50 g, 1.63 mmol), 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (779 mg, 1.63 mmol), and potassium carbonate (6.78 g, 49.0 mmol) were dissolved in dioxane (80 mL) and water (20 mL), and the reaction mixture was stirred at 95 °C for 12 h. After completion of the reaction, the reaction mixture was directly filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 15 / 1 - 1 / 1, V / V) to obtain compound A-3. MS-ESI calculated value [M+H] + 281, found 281.

[0086] The second step

[0087] Compound A-3 (3 g, 9.55 mmol) was dissolved in tetrahydrofuran (32 mL), water (8 mL), and methanol (8 mL). Lithium hydroxide monohydrate (1.20 g, 28.7 mmol) was added, and the mixture was stirred at 28 °C for 12 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran and methanol. Water (100 mL) was added for dilution, and the mixture was extracted with methyl tert-butyl ether (60 mL×1). The aqueous phase was adjusted to pH 3 with dilute hydrochloric acid aqueous solution (1 M), filtered by suction, and the filter cake was washed with water and dried to obtain compound intermediate A. MS-ESI calculated value [M+H] + 253, found 253.

[0088] Intermediate B

[0089]

[0090] Synthetic route:

[0091]

[0092] The first step

[0093] Hexamethylenetetramine (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 mixture was stirred at 30 °C for 1 h. The reaction mixture was filtered, and the filter cake was collected to obtain crude compound B-2.

[0094] The second step

[0095] Compound B-2 (65.0 g, 176 mmol) was dissolved in ethanol (429 mL), and then 35% concentrated hydrochloric acid (52.0 mL) was added. The reaction mixture was stirred at 30 °C for 72 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the hydrochloride of compound B-3.

[0096] The third step

[0097] The hydrochloride of compound B-3 (30 g, 149 mmol) was dissolved in water (375 mL), and then sodium bicarbonate (31.2 g, 372 mmol), methanol (375 mL) and di-tert-butyl dicarbonate (48.7 g, 223 mmol, 51.3 mL) were added. The reaction mixture was stirred at 30 °C for 1 hour. The reaction mixture was extracted with ethyl acetate (600 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product obtained by concentration under reduced pressure was separated and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 1 / 0 - 5 / 1, V / V) to obtain compound B-4.

[0098] The fourth step

[0099] Ammonium acetate (116 g, 1.51 mol) and sodium cyanoborohydride (9.47 g, 151 mmol) were added to a solution of compound B-4 (40.0 g, 151 mmol) in methanol (400 mL). The reaction mixture was stirred at 50 °C for 18 hours. The reaction mixture was concentrated under reduced pressure, dichloromethane (600 mL) was added to the residue and stirred, filtered, and the filtrate was concentrated under reduced pressure. The crude product obtained by concentration under reduced pressure was purified using a reversed-phase preparative chromatography column (ammonia system) to obtain intermediate B. MS-ESI calculated value [M+H] + 267, found 267.

[0100] Intermediate C

[0101]

[0102] Synthetic route:

[0103]

[0104] The first step

[0105] Compound C-1 (8.00 g, 40.6 mmol) was dissolved in N,N-dimethylacetamide (80 mL). Under a nitrogen atmosphere, 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane bis(tetrafluoroborate) (28.8 g, 81.2 mmol) was added to the reaction solution, and the reaction solution was stirred at 60 °C for 14 hours. The reaction solution was concentrated under reduced pressure, and the obtained crude product was separated and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 50 / 1 - 25 / 1, V / V) to obtain compound C-2. MS-ESI calculated values [M+H] + 215 and 217, found 215 and 217.

[0106] The second step

[0107] Compound C-2 (2.30 g, 10.7 mmol) and bis(pinacolato)diboron (5.43 g, 21.4 mmol) were dissolved in dioxane (50 mL). [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (873 mg, 1.07 mmol) and potassium acetate (2.10 g, 21.4 mmol) were added, and the reaction solution was stirred at 95 °C for 14 hours. The reaction solution was concentrated under reduced pressure, and the obtained crude product was separated and purified by column chromatography (eluent: petroleum ether / ethyl acetate, 30 / 1 - 10 / 1, V / V) to obtain intermediate C. MS-ESI calculated value [M+H] + 263, found 263.

[0108] Example 1

[0109]

[0110] Synthetic route:

[0111]

[0112] The first step

[0113] Compound 1-1 (6.00 g, 39.7 mmol) was dissolved in dichloromethane (20 mL). N-Bromosuccinimide (7.06 g, 39.7 mmol) and p-toluenesulfonic acid (1.37 g, 7.94 mmol) were added, and the reaction solution was stirred at 50 °C for 12 hours. The reaction solution was diluted with water (150 mL), extracted with ethyl acetate (250 mL × 2), and the organic phases were combined. The organic phase was washed once with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product after concentration under reduced pressure was separated and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 100 / 1 - 20 / 1, V / V) to obtain compound 1-2.

[0114] The second step

[0115] Add hexamethylenetetramine (2.68 g, 19.1 mmol) to a solution of compound 1-2 (4.40 g, 19.1 mmol) in dichloromethane (60 mL). The reaction mixture was stirred at 20 °C for 1 hour. The reaction mixture was filtered, and the filter cake was collected to obtain crude compound 1-3.

[0116] The third step

[0117] Dissolve compound 1-3 (3.00 g, 8.17 mmol) in ethanol (30 mL), then add 35% concentrated hydrochloric acid (2.41 mL). The reaction mixture was stirred at 80 °C for 2 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain the hydrochloride salt of compound 1-4.

[0118] The fourth step

[0119] Dissolve the hydrochloride salt of compound 1-4 (2.80 g, 13.8 mmol) in water (25 mL) and tetrahydrofuran (25 mL), then add di-tert-butyl dicarbonate (2.41 g, 11.1 mmol, 2.54 mL) and sodium bicarbonate (3.58 g, 41.5 mmol). The reaction mixture was stirred at 20 °C for 1 hour. The reaction mixture 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 after concentration 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.

[0120] The fifth step

[0121] Add ammonium acetate (409 mg, 5.30 mmol) and sodium cyanoborohydride (33.3 mg, 530 mmol) to a solution of compound 1-5 (300 mg, 530 mmol) in methanol (10 mL). The reaction mixture was stirred at 50 °C for 1 hour. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by preparative high performance liquid chromatography (column: Waters Xbridge 150 x 25 mm x 5 μm; mobile phase: [10 mM ammonium bicarbonate aqueous solution - acetonitrile]; B%: 20% - 50%, 10 minutes) to obtain compound 1-6. MS-ESI calculated value [M+H] + 268, found 268.

[0122] The sixth step

[0123] The chemical intermediate A (151 mg, 599 μmol) was dissolved in N,N-dimethylformamide (8 mL), and then 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (296 mg, 778 μmol), N,N-diisopropylethylamine (232 mg, 1.80 mmol, 313 μL) and compound 1-6 (160 mg, 599 μmol) were added. The reaction mixture was stirred at 20 °C for 12 hours. The reaction mixture was diluted with water (80 mL), extracted with ethyl acetate (100 mL x 2), and the organic phases were combined. The organic phase was washed once with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by thin-layer chromatography (eluent: ethyl acetate) to obtain compound 1-7.

[0124] The seventh step

[0125] Compound 1-7 (220 mg, 420 μmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added, and the reaction mixture was stirred at 20 °C for 1 hour. The reaction mixture was concentrated under reduced pressure, and the residue was separated and purified by preparative high performance liquid chromatography (column: Phenomenex Synergi C18 150 x 30 mm x 4 μm; mobile phase: [0.05% aqueous hydrochloric acid - acetonitrile]; B%: 20% - 40%, 10 minutes) to obtain the hydrochloride of compound 1.

[0126] MS-ESI calculated value [M+H] + 402, found 402. 1 H NMR (400 MHz, DMSO-d6) δ 9.31 (d, J = 8.0 Hz, 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, 8.8 Hz, 1H), 7.72 (dd, J = 7.6, 8.0 Hz, 1H), 7.63 - 7.53 (m, 2H), 7.06 (d, J = 7.3 Hz, 1H), 6.76 (d, J = 8.0 Hz, 1H), 5.48 - 5.37 (m, 1H), 3.90 (s, 3H), 3.61 - 3.24 (m, 2H), 2.56 (s, 3H) ppm.

[0127] Example 2

[0128]

[0129] Synthetic route:

[0130]

[0131] The first step

[0132] Dissolve compound 2-1 (1 g, 7.29 mmol) and tert-butylsulfinamide (928 mg, 7.66 mmol) in dichloromethane (10 mL), add cesium carbonate (4.75 g, 14.6 mmol), and stir the reaction mixture at 20 °C for 15 hours. Filter the reaction mixture through diatomaceous earth, and concentrate the filtrate under reduced pressure to obtain the crude compound 2-2. MS-ESI calculated value [M+H] + 241, found 241.

[0133] The second step

[0134] Dissolve nitromethane (3.55 mL, 65.8 mmol) in dimethyl sulfoxide (15 mL), add sodium tert-butoxide (160 mg, 1.66 mmol), stir the reaction mixture at 30 °C for 1 hour, add compound 2-2 (2 g, 8.32 mmol), and continue to stir the reaction mixture at 30 °C for 15 hours. Add saturated ammonium chloride aqueous solution (100 mL) to the reaction mixture, extract with ethyl acetate (30 mL x 4), and combine the organic phases. Dry the organic phases over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain the crude compound 2-3. MS-ESI calculated value [M+H] + 302, found 302.

[0135] The fourth step

[0136] Dissolve compound 2-3 (2 g, 6.64 mmol) in ethyl acetate (3 mL), add 4M hydrogen chloride in ethyl acetate solution (13.3 mL), and stir the reaction mixture at 15 °C for 16 hours. Concentrate the reaction mixture under reduced pressure, add ethyl acetate (50 mL) to the concentrated crude product, stir for 0.5 hour, and filter to obtain the hydrochloride salt of compound 2-4.

[0137] The fifth step

[0138] Dissolve the hydrochloride salt of compound 2-4 (250 mg, 1.07 mmol), intermediate A (270 mg, 1.07 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (410 mg, 2.14 mmol), and 1-hydroxybenzotriazole (217 mg, 1.60 mmol) in tetrahydrofuran (4 mL) and N,N-dimethylformamide (1 mL), add N,N-diisopropylethylamine (553 mg, 4.28 mmol, 745 μL) to the reaction mixture. Stir the reaction mixture at 30 °C for 1 hour. Concentrate the reaction mixture under reduced pressure, and separate the obtained crude product by thin layer chromatography (eluent: dichloromethane / methanol, 10 / 1, V / V) to obtain compound 2-5. MS-ESI calculated value [M+H]+ 432, measured value 432.

[0139] The sixth step

[0140] Dissolve compound 2-5 (120 mg, 220 μmol) in ethanol (2 mL) and water (0.5 mL), add iron powder (77.7 mg, 1.39 mmol) and ammonium chloride (149 mg, 2.78 mmol), and stir the reaction solution at 95 °C for 2 hours. The reaction solution was concentrated under pressure, the residue was added to dichloromethane / methanol (10 / 1, V / V) solution (50 mL), filtered, and the filtrate was concentrated under reduced pressure. The crude product obtained by concentration under reduced pressure was separated and purified by preparative high performance liquid chromatography (column: Xtimate C18 150x 25mm x 5μm; mobile phase: [0.05% aqueous ammonium bicarbonate solution - acetonitrile]; B%: 21% - 41%, 10 minutes) to obtain compound 2.

[0141] MS-ESI calculated value [M+H] + 402, measured value 402. 1 H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 6.0 Hz, 1H), 8.24 (s, 1H), 8.05 (s, 1H), 7.93 - 7.81 (m, 2H), 7.74 (dd, J = 1.6, 8.8 Hz, 1H), 7.61 - 7.54 (m, 2H), 7.04 - 6.94 (m, 1H), 6.87 (dd, J = 2.4, 5.6 Hz, 1H), 5.14 - 4.93 (m, 1H), 3.82 (s, 3H), 3.17 - 2.92 (m, 2H), 2.56 (s, 3H) ppm.

[0142] Example 3

[0143]

[0144] Synthetic route:

[0145]

[0146] The first step

[0147] Refer to the first step of Example 2 to obtain compound 3-2. MS-ESI calculated value [M+H] + 211, measured value 211.

[0148] The second step

[0149] Refer to the second step of Example 2 to obtain compound 3-3.

[0150] The third step

[0151] The hydrochloride salt of compound 3-4 was obtained in the third step of Reference Example 2.

[0152] The fourth step

[0153] Compound 3-5 was obtained in the fourth step of Reference Example 2. MS-ESI calculated value [M+H] + 402, found 402.

[0154] The fifth step

[0155] The crude product was obtained in the fifth step of Reference Example 2. The obtained crude product was separated and purified by preparative high performance liquid chromatography (column: Phenomenex Synergi C18 150x 30mm x 4μm; mobile phase: [0.05% hydrochloric acid aqueous solution - acetonitrile]; B%: 7% - 27%, 11 minutes) to obtain the hydrochloride salt of compound 3.

[0156] MS-ESI calculated value [M+H] + 372, found 372. 1 H NMR (400 MHz, DMSO-d6) δ 9.92 (d, J = 7.6 Hz, 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 = 8.4 Hz, 2H), 7.83 (d, J = 8.8 Hz, 1H), 7.66 - 7.47 (m, 2H), 5.76 - 5.59 (m, 1H), 3.74 - 3.39 (m, 2H), 2.58 (s, 3H) ppm.

[0157] Example 4

[0158]

[0159] Synthetic route:

[0160]

[0161] The first step

[0162] Compound 4-2 was obtained in the first step of Reference Example 2. MS-ESI calculated value [M+H] + 241, found 241.

[0163] The second step

[0164] Compound 4-3 was obtained in the second step of Reference Example 2. MS-ESI calculated value [M+H] + 302, found 302.

[0165] The third step

[0166] The hydrochloride of compound 4-4 was obtained in the third step of Reference Example 2.

[0167] The fourth step

[0168] Compound 4-5 was obtained in the fourth step of Reference Example 2. MS-ESI calculated value [M+H] + 432, found 432.

[0169] The fifth step

[0170] Compound 4 was obtained in the fifth step of Reference Example 2.

[0171] MS-ESI calculated value [M+H] + 402, found 402. 1 H NMR (400 MHz, DMSO-d6) δ 12.71 (s, 1H), 8.85 (d, J = 8.0 Hz, 1H), 8.28 - 8.15 (m, 3H), 8.04 (s, 1H), 7.93 - 7.81 (m, 2H), 7.72 (dd, J = 1.6, 8.4 Hz, 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.

[0172] Example 5

[0173]

[0174] Synthetic route:

[0175]

[0176] The first step

[0177] Compound 5-2 was obtained in the sixth step of Reference Example 1. MS-ESI calculated value [M+H] + 451, 453, found 451, 453.

[0178] The second step

[0179] Compound 5-2 (60.0 mg, 133 μmol), compound 5-3 (41.2 mg, 160 μmol), and potassium carbonate (36.8 mg, 266 μmol) were added to dioxane (4.5 mL) and water (0.5 mL), and then [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (9.73 mg, 13.3 μmol) was added. The reaction mixture was stirred at 90 °C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel column chromatography (eluent: dichloromethane / methanol, 200 / 1 - 10 / 1, V / V) to obtain compound 5-4. MS-ESI calculated value [M+H] + 503, found 503.

[0180] The third step

[0181] The hydrochloride salt of compound 5 was obtained according to the seventh step of Reference Example 1.

[0182] MS-ESI calculated value [M+H] + 403, found 403. 1 H NMR (400 MHz, DMSO-d6) δ 9.56 (d, J = 8.8 Hz, 1H), 8.75 (d, J = 5.2 Hz, 1H), 8.56 - 8.42 (m, 1H), 8.38 - 8.27 (m, 1H), 8.21 - 8.03 (m, 3H), 7.85 (d, J = 8.4 Hz, 1H), 7.74 (t, J = 7.6 Hz, 1H), 7.62 (d, J = 8.8 Hz, 1H), 7.06 (d, J = 7.6 Hz, 1H), 6.80 (d, J = 8.0 Hz, 1H), 5.54 - 5.40 (m, 1H), 3.92 (s, 3H), 3.58 - 3.42 (m, 2H), 2.64 - 2.54 (m, 3H) ppm.

[0183] Example 6

[0184]

[0185] Synthetic route:

[0186]

[0187] The first step

[0188] Compound 6-1 (1 g, 3.80 mmol) was dissolved in N,N-dimethylformamide (8 mL), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.12 g, 5.85 mmol), 1-hydroxybenzotriazole (579 mg, 4.29 mmol) and ammonia water (1.82 g, 13.0 mmol, 2.00 mL, 25% purity) were added. The reaction mixture was stirred at 20 °C for 15 h. Water (60 mL) was added to the reaction mixture, and the mixture was stirred at 0 °C for 1 h, filtered, and the filter cake was collected to obtain crude compound 6-2.

[0189] The second step

[0190] Compound 6-2 (470 mg, 1.79 mmol) was added to trimethoxymethane (4.84 g, 45.6 mmol, 5 mL) and N-methylpyrrolidone (1 mL), and p-toluenesulfonic acid (34.1 mg, 179 μmol) was further added. The reaction mixture was stirred at 110 °C for 12 h. Water (60 mL) was added to the reaction mixture, and the mixture was continuously stirred at 0 °C for 1 h. The reaction mixture was filtered, and the filter cake was collected to obtain crude compound 6-3.

[0191] The third step

[0192] Compound 6-3 (370 mg, 1.36 mmol) was added to N,N-dimethylformamide (0.1 mL), and thionyl chloride (4 mL) was further added. The reaction mixture was stirred at 90 °C for 14 h. After the reaction mixture was cooled to room temperature, it was concentrated under reduced pressure. Toluene (20 mL) was added to the residue obtained by concentration under reduced pressure, and it was washed once with saturated sodium bicarbonate (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude compound 6-4.

[0193] MS-ESI calculated value [M+H] + 291, found 291.

[0194] The fourth step

[0195] Compound 6-4 (220 mg, 757 μmol) and intermediate B (205 mg, 757 μmol) were dissolved in isopropanol (5 mL), and sodium carbonate (161 mg, 1.51 mmol) was added. The reaction mixture was stirred at 60 °C for 15 h. The reaction mixture was concentrated under reduced pressure, and the obtained crude product was separated and purified by column chromatography (eluent: petroleum ether / ethyl acetate, 10 / 1 - 4 / 1, V / V) to obtain compound 6-5.

[0196] MS-ESI calculated value [M+H] + 521, found 521.

[0197] The fifth step

[0198] Compound 6-5 (200 mg, 384 μmol), intermediate C (101 mg, 384 μmol), potassium phosphate (163 mg, 769 μmol) and dichloromethane complex of [1,1′-bis(diphenylphosphino)ferrocene] palladium(II) dichloride (62.8 mg, 76.9 μmol) were added to N,N-dimethylformamide (3 mL) and water (0.3 mL). The reaction mixture was stirred at 85 °C for 16 hours. The reaction mixture was concentrated under reduced pressure, and the resulting crude product was separated and purified by column chromatography (eluent: petroleum ether / ethyl acetate, 10 / 1 - 1 / 1, V / V) to obtain compound 6-6.

[0199] MS-ESI calculated value [M+H] + 529, found 529.

[0200] The sixth step

[0201] Compound 6-6 (75 mg, 142 μmol) was added to 4M hydrogen chloride in ethyl acetate solution (5 mL). The reaction mixture was stirred at 20 °C for 40 minutes. The reaction mixture was concentrated under reduced pressure, and the resulting crude product was separated and purified by preparative high performance liquid chromatography (column: Phenomenex Synergi C18 150x 25mm 10μm; mobile phase: [0.05% hydrochloric acid aqueous solution - acetonitrile]; B%: 8% - 28%, 10 minutes) to obtain the hydrochloride salt of compound 6.

[0202] MS-ESI calculated value [M+H] + 429, found 429. 1 1H NMR (400 MHz, DMSO-d6) δ 13.04 - 12.67 (m, 1H), 9.42 - 9.06 (m, 1H), 8.68 (s, 1H), 8.59 - 8.44 (m, 3H), 8.01 (d, J = 7.2 Hz, 1H), 7.89 - 7.80 (m, 2H), 7.67 (d, J = 8.8 Hz, 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 = 6.4 Hz, 1H), 4.09 - 3.82 (m, 1H), 3.79 - 3.70 (m, 3H), 3.45 - 3.21 (m, 1H) ppm.

[0203] Example 7

[0204]

[0205] Synthesis route:

[0206]

[0207] The first step

[0208] Compound 7-1 was obtained by referring to the fifth step of Example 6.

[0209] Calculated value of MS-ESI [M+H] + 525, measured value 525.

[0210] The second step

[0211] The hydrochloride salt of compound 7 was obtained by referring to the sixth step of Example 6.

[0212] Calculated value of MS-ESI [M+H] + 425, measured value 425. 1 1H NMR (400 MHz, DMSO-d6) δ 11.42 (d, J = 7.6 Hz, 1H), 9.29 (d, J = 8.4 Hz, 1H), 8.69 (s, 1H), 8.58 (s, 3H), 8.02 (dd, J = 1.0, 7.6 Hz, 1H), 7.92 - 7.82 (m, 2H), 7.65 (d, J = 8.4 Hz, 1H), 7.42 (dd, J = 1.6, 8.4 Hz, 1H), 7.34 - 7.27 (m, 2H), 7.18 (d, J = 7.8 Hz, 1H), 6.90 (dd, J = 2.0, 8.0 Hz, 1H), 6.10 - 5.93 (m, 1H), 4.10 - 3.87 (m, 1H), 3.76 (s, 3H), 3.35 (dd, J = 4.0, 7.2 Hz, 1H), 2.52 (s, 3H) ppm.

[0213] Example 8

[0214]

[0215] Synthesis route:

[0216]

[0217] The first step

[0218] Compound 8-1 (4.4 g, 17.3 mmol) was dissolved in dichloromethane (40 mL). The reaction solution was cooled to 0 °C, and m-chloroperoxybenzoic acid (5.25 g, 25.9 mmol, 85% purity) was added portionwise. The reaction solution was stirred at 20 °C for 15 hours. The reaction solution was diluted with dichloromethane (100 mL) and washed successively with 10% aqueous sodium bicarbonate solution (100 mL), 10% aqueous sodium thiosulfate solution (100 mL), and saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product obtained by concentration was separated and purified by silica gel column chromatography (eluent: dichloromethane / methanol, 100 / 1 - 10 / 1, V / V) to obtain compound 8-2.

[0219] MS-ESI calculated value [M+H] + 272, found 272.

[0220] The second step

[0221] Compound 8-2 (1 g, 3.69 mmol) was dissolved in dichloromethane (20 mL), triethylamine (747 mg, 7.38 mmol, 1.03 mL) was added, and the reaction solution was cooled to -70 °C. Oxalyl chloride (937 mg, 7.38 mmol, 646 μL) was slowly added dropwise. The reaction solution was stirred at -70 °C for 0.5 hour, warmed to 20 °C, and stirred for an additional 2 hours. Methanol (5 mL) was added to dilute the reaction solution, and the resulting organic phase was washed successively with saturated sodium bicarbonate (50 mL) and water (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product obtained by concentration was separated and purified by column chromatography (eluent: petroleum ether / ethyl acetate, 100 / 1 - 50 / 1, V / V) to obtain compound 8-3.

[0222] MS-ESI calculated value [M+H] + 290, found 290.

[0223] The third step

[0224] Compound 8-4 was obtained by referring to the fifth step of Reference Example 6.

[0225] MS-ESI calculated value [M+H] + 298, found 298.

[0226] The fourth step

[0227] Compound 8-5 was obtained by referring to the fourth step of Reference Example 6.

[0228] MS-ESI calculated value [M+H] + 528, found 528.

[0229] The fifth step

[0230] Refer to the sixth step of Example 6 to obtain the hydrochloride salt of Compound 8.

[0231] MS-ESI calculated value [M+H] + 428, found 428. 1 H NMR (400 MHz, DMSO-d6) δ 10.33 (s, 1H), 9.31 (d, J = 8.4 Hz, 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, 7.6 Hz, 2H), 7.45 - 7.26 (m, 4H), 7.08 (d, J = 7.2 Hz, 1H), 6.92 (d, J = 7.6 Hz, 1H), 5.85 - 5.75 (m, 1H), 4.02 - 3.85 (m, 1H), 3.77 (s, 3H), 3.47 - 3.35 (m, 1H) ppm.

[0232] Example 9

[0233]

[0234] Synthetic route:

[0235]

[0236] The first step

[0237] Refer to the fifth step of Example 6 to obtain Compound 9-1.

[0238] MS-ESI calculated value [M+H] + 294, found 294.

[0239] The second step

[0240] Refer to the fourth step of Example 6 to obtain Compound 9-2.

[0241] MS-ESI calculated value [M+H] + 524, found 524.

[0242] The third step

[0243] Refer to the sixth step of Example 6 to obtain the hydrochloride salt of Compound 9.

[0244] MS-ESI calculated value [M+H] + 424, found 424. 11H NMR (400 MHz, DMSO-d6) δ 12.81 (d, J = 7.6 Hz, 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 = 5.6 Hz, 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.

[0245] Activity Test

[0246] 1. In vitro evaluation of ROCK protein kinase inhibitory activity

[0247] Experimental purpose: To detect the ROCK protein kinase inhibitory IC 50 value.

[0248] Experimental materials:

[0249] Assay buffer: 20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.5), 10 mM magnesium chloride, 1 mM ethylene glycol bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid, 0.02% Brij 35, 0.02 mg / mL bovine serum albumin, 0.1 mM sodium orthovanadate, 2 mM dithiothreitol, 1% DMSO. Experimental procedure:

[0250] Add the ROCK2 protein kinase substrate Long S6 Kinase substrate peptide at a concentration of 20 μM to the freshly prepared buffer, and then add 1 nM ROCK2 protein kinase and stir evenly. Use an Echo 550 to add a series of DMSO dilutions (starting from 10 μM, diluted in a 3-fold series) containing the compound to be tested, pre-incubate at room temperature for 20 minutes, and add 33 P-ATP (radioactivity 10 μCi / μL) to initiate the reaction, and react at room temperature for two hours. Then filter using a P81 ion exchange paper (Whatman #3698-915) and wash with 0.75% phosphoric acid. Detect the radioactivity using the Filter-Binding method.

[0251] The protein kinase inhibitory activity of the compound is expressed as the remaining protein kinase activity relative to the blank substrate (pure DMSO). Use the Prism software package (GraphPad Software, San Diego, California, USA) to calculate the IC 50 value and the curve.

[0252] Experimental results:

[0253] Table 1 Test results of ROCK inhibitory activity

[0254] Compound <![CDATA[IC for ROCK2 50 (nM)]]> 1 (hydrochloride) 8 3 (hydrochloride) 45 4 54 5 (hydrochloride) 27 6 (hydrochloride) 56 7 (hydrochloride) 50 8 (hydrochloride) 63 9 (hydrochloride) 72

[0255] The compounds of the present invention have significant and even unexpected ROCK2 inhibitory activity.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein, T1 is CH; T3 is CH; Structural unit is T4 is N or CH; R1 is H, F, Cl or C 1-3 alkyl; R2, R3 and R4 are H; R6 is -CH2NH2 and R7 is H; Each R8 is independently H, F, Cl, C 1-3 alkyl or C 1-3 alkoxy; n is 1 or 2.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound has the formula (I-16): Among them, R1, R8 and n are as defined in claim 1; the carbon atom marked with "*" is a chiral carbon atom and exists in the form of a single (R) or (S) enantiomer or an enantiomer-rich form.

3. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound has the formula (I-17): Among them, R1, R8 and n are as defined in claim 1; the carbon atom marked with "*" is a chiral carbon atom and exists in the form of a single (R) or (S) enantiomer or an enantiomer-rich form.

4. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein R1 is F or -CH3.

5. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein each R8 is independently H or -OCH3.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit is 7. The compound or a pharmaceutically acceptable salt thereof according to claim 6, wherein the structural unit is 8. A compound of the following formula or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

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