Azacycloalkane compounds, pharmaceutical compositions and uses thereof

Abstract

The present application relates to an azacycloalkane compound having the following structural formula I, a pharmaceutical composition, and uses thereof. The azacycloalkane compound and pharmaceutical composition thereof provided by the present invention have significant Rho kinase inhibitory activity, and their enzymatic and cellular activities are superior to those of existing ROCK inhibitors such as ripasudil, netarsudil, and vermosudil, and have great applicability. JPEG2026504901000133.jpg166156

Description

[Technical Field]

[0001] The present application provides azacycloalkane compounds, stereoisomers containing the compounds, pharmaceutical compositions, and uses of the compounds and pharmaceutical compositions. The azacycloalkane compounds are inhibitors that target ROCK and can be used to regulate diseases mediated by Rho kinase. [Background technology]

[0002] The Rho kinase / ROCK (Rho-associated kinase) signaling pathway induces cytoskeletal reorganization, cell migration, adhesion, and stress fiber formation, and is involved in various physiological functions. The ROCK family consists of ROCK1 and ROCK2. ROCK1 is highly expressed in the lung, liver, spleen, kidney, and testis, while ROCK2 is highly expressed in the brain and heart. ROCK1 indirectly interacts with the cadherin-catenin complex by binding to p120-catenin, a scaffolding protein of E-cadherin. ROCK1 can concentrate at the center of microtubule organization and at the edges of pseudopodia in motile cells, and is involved in cell migration. ROCK2 is primarily localized in the cytoplasm and localizes to the plasma membrane via its C-terminal domain, where it associates with vimentin and actin stress fibers.

[0003] ROCK mediates numerous pathophysiological signals and is involved in various physiological functions, including endothelial permeability, tissue contraction, and growth. ROCK inhibitors have potential applications in diseases such as asthma, cancer, glaucoma, insulin resistance, renal failure, neurodegeneration, and osteoporosis. Two ROCK1 / ROCK2 inhibitors, ripasudil and netarsudil, are approved for the treatment of glaucoma, and one ROCK2 inhibitor, belmosudil, is approved for the treatment of chronic graft-versus-host disease (cGVHD). Summary of the Invention [Problem to be solved by the invention]

[0004] However, the activity of currently available ROCK inhibitors is weak, and the functional and clinical value of ROCK is not fully realized. Therefore, there is an urgent need to develop drugs that target ROCK to address unmet clinical needs. [Means for solving the problem]

[0005] In this application, highly active azacycloalkane compounds targeting ROCK with a novel scaffold and pharmaceutical compositions thereof are developed for use in modulating Rho kinase-mediated diseases.

[0006] One aspect of the present invention provides an azacycloalkane compound of Formula I, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled form, isomer, or prodrug thereof: JPEG2026504901000002.jpg166156 Yet another aspect of the present invention provides methods for preparing compounds of Formula I, isotopic isomers, or pharmaceutically acceptable salts, solvates, active metabolites, crystalline polymorphs, isotopically labeled forms, isomers, or prodrugs thereof, and their use for treating diseases mediated by ROCK. [Brief explanation of the drawings]

[0007] [Figure 1] Figure 1 shows the changes in body weight of animals in each group during the test period. [Figure 2] Figure 2 shows the 24-hour intraocular pressure change curves of animals in each group at different times after administration at D1. [Figure 3] FIG. 3 shows the results of RGCs from animals in each group at the end of the experiment. DETAILED DESCRIPTION OF THE INVENTION

[0008] The present application will be described in more detail through the following embodiments, through which the features and advantages of the present application will be more clearly defined.

[0009] The word "exemplary" is used herein to mean "serving as an example, embodiment, or illustration." An embodiment described herein as "exemplary" should not be construed as superior or advantageous over other embodiments.

[0010] Furthermore, the technical features in different embodiments of the present application described below can be combined with each other if they are not contradictory.

[0011] The following embodiments are provided for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art should readily recognize that a variety of non-critical parameters can be changed or modified to achieve substantially the same results.

[0012] definition Unless otherwise defined, all technical terms in this specification have the same meaning as commonly understood by those skilled in the art to which the claimed subject matter belongs. Unless otherwise specified, all patents, patent applications, and disclosure materials cited throughout this specification are incorporated herein by reference in their entirety. When a trade name is mentioned in this specification, it is intended to refer to that product or active ingredient.

[0013] It is to be understood that the foregoing brief description and the following detailed description are exemplary and explanatory only and are not intended to limit the subject matter of the present invention in any way. It is important to note that, as used in the specification and claims, the singular forms "a," "an," and "the" include the plural of the referenced item unless the context clearly dictates otherwise. It is also to be noted that, unless otherwise indicated, the terms "or" and "alternative" are used to indicate "and / or." Furthermore, the use of the term "including," such as "comprising," "containing," and "containing," is not limiting.

[0014] Definitions of standard chemical terms are found in Carey and Sundberg's "Advanced Organic Chemistry 4 th Reference can be made to literature such as "The Journal of Chemistry, Vol. A (2000) and B (2001), Plenum Press, New York." Unless otherwise specified, conventional methods within the skill of the art, such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacological methods, are used. Unless specific definitions are provided, the nomenclature, laboratory procedures, and techniques associated with analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry used herein are known to those skilled in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, drug delivery, and patient treatment. Standard techniques can be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipid transfection). For example, reactions and purification techniques can be performed using kits accompanied by manufacturer-provided instructions, according to methods known in the art, or according to the methods described herein. In general, the techniques and steps can be performed by conventional methods well known in the art and described in various general or more specific publications. Such documents are cited and discussed in this application.

[0015] When a substituent is represented by a conventional chemical formula written from left to right, the substituent also includes the chemically equivalent substituent that results when the structural formula is written from right to left. For example, CHO is equivalent to OCH.

[0016] The term "substituted or unsubstituted" encompasses both "substituted" and "unsubstituted," where "substituted" means that any one or more hydrogen atoms on a given atom are replaced with a substituent, provided that the valence of the given atom is normal and the substituted compound is stable, and "unsubstituted" means that no hydrogen atoms on a given atom are replaced with a substituent. For example, "substituted or unsubstituted ethyl" (e.g., where the substituent is a halogen) includes unsubstituted (-CH2CH3), monosubstituted (e.g., -CH2CH2F), polysubstituted (e.g., -CHFCH2F, -CH2CHF2, etc.), or fully substituted (-CF2CF3). As will be understood by those skilled in the art, any group containing one or more substituents will not introduce a substituent or substitution pattern that is spatially impossible and / or cannot be synthesized. When a substituent is an oxo group (i.e., =0), this means that two hydrogen atoms on the same atom are replaced.

[0017] When any variable (e.g., R) occurs more than once in any composition or structure of a compound, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0 to 2 R, then that group may optionally be substituted with up to 2 R, and each occurrence of R is an independent option. Furthermore, combinations of substituents and / or variants thereof are permissible only if such combinations result in stable compounds. The term "optionally" or "may" means that the subsequently described event or circumstance may or may not occur, and the description includes both the occurrence and non-occurrence of the event or circumstance.

[0018] As used herein, C m~n means that the moiety has m to n carbon atoms. For example, 1~8 " group has 1 to 8 carbon atoms in the moiety, i.e., the group is said to contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, ... 8 carbon atoms. Thus, for example, "C 1~8"Alkyl" refers to an alkyl group containing 1 to 8 carbon atoms. That is, the alkyl is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl...octyl, etc. Numeric ranges herein, such as "1 to 8," refer to each integer within the range; for example, "1 to 8 carbon atoms" means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, or 8 carbon atoms.

[0019] The term "alkyl" refers to an optionally substituted straight-chain or an optionally substituted branched-chain saturated aliphatic hydrocarbon group connected to the rest of the molecule by a single bond. As used herein, "alkyl" can have from 1 to about 8 carbon atoms, such as from 1 to 6 carbon atoms, or from 1 to 4 carbon atoms, or from 1 to 3 carbon atoms. Examples of "alkyl" as used herein include, but are not limited to, alkyl groups such as methyl, ethyl, n-propyl, isopropyl, 2-methyl-l-propyl, 2-methyl-2-propyl, 2-methyl-l-butyl, 3-methyl-l-butyl, 2-methyl-3-butyl, 2,2-dimethyl-l-propyl, 2-methyl-l-pentyl, 3-methyl-l-pentyl, 4-methyl-l-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-l-butyl, 3,3-dimethyl-l-butyl, 2-ethyl-l-butyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, iso-pentyl, neo-pentyl, tert-pentyl, hexyl, and longer alkyl groups such as heptyl and octyl. When a numerical range exists for a group defined herein, e.g., "alkyl," it is preferred to use a range such as "C 1~8 "Alkyl" refers to alkyls consisting of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, and also includes, for example, "C 1~4"Alkyl" refers to alkyls composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, or 4 carbon atoms. In this specification, alkyls also include those without a specified numerical range.

[0020] The term "alkenyl" refers to an optionally substituted straight-chain or optionally substituted branched-chain monovalent hydrocarbon group having at least one C=C double bond. The alkenyl may have from 2 to about 8 carbon atoms, such as, but not limited to, from 2 to about 6 carbon atoms, or from 2 to about 4 carbon atoms. The double bond in these groups may be in either the cis or trans configuration, and both isomers should be understood to be included. Examples of alkenyl include, but are not limited to, ethylenyl (CH=CH), 1-propenyl (CHCH=CH), isopropenyl (C(CH)=CH), butenyl, and 1,3-butadienyl. When a numerical range exists for alkenyl as defined herein, for example, "C 2-8 "Alkenyl" refers to alkenyl consisting of 2, 3, 4, 5, 6, 7, or 8 carbon atoms. In this specification, alkenyl also includes those without a specified numerical range.

[0021] The term "alkynyl" refers to an optionally substituted, straight- or branched-chain monovalent hydrocarbon group having at least one C≡C triple bond. The alkynyl may have, for example, but not limited to, 2 to about 8 carbon atoms, such as 2 to about 6 carbon atoms, or 2 to about 4 carbon atoms. Examples of alkynyl herein include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and 1,3-butadinyl. When a numerical range exists for alkynyl as defined herein, for example, "C 2~8 "Alkynyl" refers to alkynyl consisting of 2, 3, 4, 5, 6, 7, or 8 carbon atoms. In this specification, alkynyl also includes those without a specified numerical range.

[0022] The term "cycloalkyl" refers to a non-aromatic carbocyclic ring, including a saturated carbocyclic ring (e.g., cycloalkyl) or an unsaturated carbocyclic ring (e.g., cycloalkenyl). Carbocyclic rings may be monocyclic carbocyclic rings (having one ring), e.g., monocyclic cycloalkyl, bicyclic carbocyclic rings (having two rings), e.g., bicyclic cycloalkyl, or polycyclic carbocyclic rings (having two or more rings). Rings may be bridged or spiro-connected. Carbocyclic rings (e.g., cycloalkyl or cycloalkenyl) may have 3 to 8 carbon atoms, e.g., 3 to about 6 ring carbon atoms or 3 to about 5 ring carbon atoms.

[0023] The term "aryl" refers to an optionally substituted aromatic hydrocarbon group having 6 to about 20, e.g., 6 to 12 or 6 to 10, ring carbon atoms, and may be a monocyclic aryl, a bicyclic aryl, or a polycyclic aryl. A bicyclic aryl or a polycyclic aryl may be a monocyclic aryl fused to another independent ring, such as an aliphatic ring, a heterocyclic ring, an aryl ring, or an aryl heterocyclic ring. Non-limiting examples of monocyclic aryls include monocyclic aryls having 6 to about 12, 6 to about 10, or 6 to about 8 ring carbon atoms, such as phenyl; an example of a bicyclic aryl is naphthyl; and examples of polycyclic aryls include phenanthryl, anthracenyl, and azulenyl.

[0024] The term "heteroaryl" refers to an optionally substituted heteroaryl containing about 5 to about 20, e.g., 5-12 or 5-10, skeletal ring atoms, wherein at least one (e.g., 1-4, 1-3, 1-2) ring atom is a heteroatom. The heteroatoms are independently selected from, but not limited to, oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin heteroatoms. Heteroaryl includes monocyclic heteroaryl (having one ring), bicyclic heteroaryl (having two rings), or polycyclic heteroaryl (having two or more rings). In embodiments where two or more heteroatoms are present in a ring, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different from each other. A bicyclic heteroaryl or higher polycyclic heteroaryl may be a monocyclic heteroaryl fused to another independent ring, such as an aliphatic ring, a heterocyclic ring, an aryl ring, or an aryl heterocyclic ring (collectively referred to as a fused cycloheteroaryl). Non-limiting examples of heteroaryl include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolyl, isoquinolyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothiophenyl, indolyl, isoindolyl, and the like.

[0025] The terms "heterocyclyl" and "heterocyclic group" refer to non-aromatic heterocycles, including saturated or unsaturated heterocycles (containing unsaturated bonds) that do not have a completely conjugated π-electron system, and are classified as non-aromatic monocyclic, fused polycyclic, bridged, or spiro rings. Among these, one or more (e.g., 1 to 4, 1 to 3, or 1 to 2) of the ring atoms are heteroatoms, such as oxygen, nitrogen, or sulfur atoms. Heterocycles include monocyclic heterocycles (having one ring), bicyclic heterocycles (having two bridged rings), and polycyclic heterocycles (having two or more bridged rings), including spiro rings. A heterocyclyl may have 3 to about 20, e.g., 3 to about 10, 3 to about 8, 4 to 8, 4 to 7, 5 to about 8, or 5 to about 6 ring atoms. Non-limiting examples of heterocyclyl include oxiranyl, thioethanethiol, aziridinyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, pyrrolidinyl, oxazolidinyl, tetrahydropyrazolyl, pyrrolinyl, dihydrofuranyl, dihydrothienyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, dihydropyridyl, tetrahydropyridyl, dihydropyranyl, tetrahydropyranyl, dihydrothiopyranyl, azepanyl, oxetanyl, thiepanyl, oxazabicyclo[2.2.1]heptyl, and azapyro[3.3]heptyl, and the like.

[0026] The term "halo" or "halogen" refers to an optionally substituted group (e.g., alkyl, alkenyl, alkynyl, alkoxy, etc.) in which at least one hydrogen atom is replaced with a halogen (e.g., fluorine, chlorine, bromine, iodine, or a combination thereof). In some embodiments, two or more hydrogens are replaced with halogens that are identical to each other (e.g., difluoromethyl, trifluoromethyl); in other embodiments, two or more hydrogens are replaced with halogens that are not identical to each other (e.g., 1-chloro-1-fluoro-1-iodoethyl).

[0027] The term "alkoxy" refers to an alkyl ether (O-alkyl), and non-limiting examples of alkoxy include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

[0028] The term "alkyl acyl" refers to a group having an alkyl group attached to -CO-, and non-limiting examples of the term include formyl, acetyl, propanoyl, butyryl, and the like. For example, the term "C 1~6 "Alkyl acyl" means C 1-6 It refers to a group in which an alkyl group is bonded to a -CO- group. 1~4 "Alkyl acyl" means C 1~4 It refers to a group formed by bonding an alkyl group to -CO-.

[0029] The term "alkylsulfonyl" refers to an alkyl group attached to -SO2-, and non-limiting examples of the term include methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl, and the like. For example, the term "C 1~6 "Alkylsulfonyl" means -SO2- with C 1~6 It refers to a group to which an alkyl group is attached. 1-4 "Alkylsulfonyl" means -SO2- with C 1-4 It refers to a group to which an alkyl group is bonded.

[0030] The term "heteroarylacyl" refers to a group in which a heteroaryl group is attached to -CO-. For example, the term "C 5~20 "Heteroarylsulfonyl" means -CO- to C 5~20 Refers to a group to which a heteroaryl group is attached. 5~20 "Heteroaryl" is defined above.

[0031] Other radical terms used herein include "hydroxyl" which refers to an --OH group, "sulfhydryl" which refers to an --SH group, "cyano" which refers to a --CN group, and "carboxy" which refers to a --COOH group.

[0032] The term "membered" refers to the number of atoms that make up the backbone of the ring. For example, pyridine is a six-membered ring and pyrrole is a five-membered ring.

[0033] The term "pharmaceutically acceptable" means that the compound, material, composition and / or dosage form is, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals and does not cause excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0034] The term "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical component or agent, i.e., a "carrier," that facilitates the introduction of the compound into cells or tissues, including, but not limited to, stabilizers, diluents, suspending agents, thickeners, and / or excipients.

[0035] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acid and free base of a particular compound without any undesirable biological or other effects. Unless otherwise specified, salts in this invention can refer to metal salts, ammonium salts, salts formed with organic bases, salts formed with inorganic acids, salts formed with organic acids, salts formed with basic or acidic amino acids, and the like. Non-limiting examples of metal salts include, but are not limited to, alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts, magnesium salts, and barium salts; and aluminum salts. Non-limiting examples of salts formed with organic bases include, but are not limited to, salts formed with trimethylamine, triethylamine, pyridine, methylpyridine, 2,6-dimethylpyridine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexyleneamine, and the like. Non-limiting examples of salts formed with inorganic acids include, but are not limited to, salts formed with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Non-limiting examples of salts formed with organic acids include, but are not limited to, salts formed with formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, maleic acid, tartaric acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfinic acid, p-toluenesulfonic acid, etc. Non-limiting examples of salts formed with alkaline amino acids include, but are not limited to, salts formed with arginine, lysine, ornithine, etc. Non-limiting examples of salts formed with acidic amino acids include, but are not limited to, salts formed with aspartic acid, glutamic acid, etc.

[0036] Pharmaceutically acceptable salts can be synthesized from parent compounds containing acid or basic groups by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or free base form of the compound with a stoichiometrically appropriate base or acid in water or an organic solvent, or a mixture of both. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropyl alcohol, or acetonitrile are preferably used.

[0037] The term "solvate," as used herein, refers to a physical association of a compound with one or more solvent molecules, which may contain varying degrees of ionic and covalent bonding (e.g., hydrogen bonding). It has been found that such solvates can be isolated. For example, one or more solvent molecules may be present in the crystalline lattice of a crystal. The term "solvate" encompasses both solvent phases and separable solvates. Numerous examples of corresponding solvates exist, including ethanol solvates and methanol solvates. A "hydrate" is a solvate in which water (HO) molecules are the solvent. One or more compounds of the present invention may optionally be prepared as a solvate. The preparation of solvates is well known. For example, M. Caira et al., J. Pharmaceutical Sci., 93(3), 601-611 (2004), describes the preparation of a solvate of the antifungal drug fluconazole, namely, a preparation using ethyl acetate and water. Similar methods for preparing solvates and hydrates are also described in EC van Tonder et al., AAPS PharmSciTech., 5(1), article 12 (2004); and ALBingham et al., Chem. Commun., 603-604 (2001). A typical, non-limiting example of a preparation process involves dissolving a compound of the present invention in a desired amount of a desired solvent (organic solvent, water, or a mixture thereof) at room temperature or higher, cooling, and allowing the solution to stand to precipitate crystals, which are then isolated and separated by standard methods. The presence of the solvate- (hydrate-) forming solvent (water) in the crystals can be verified by IR spectroscopic techniques.

[0038] The term "active metabolite" refers to an active derivative of a compound that is produced when the compound is metabolized.

[0039] The term "polymorphs" refers to compounds of the present invention that exist in different crystalline structures.

[0040] The term "isotopically labeled compound" refers to a compound of the present invention that is labeled with an isotope. For example, the isotopes in the compound of the present invention include various isotopes of elements such as H, C, N, O, P, F, and S. 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F and 36 S is included.

[0041] The term "pharmaceutically acceptable prodrug" or "prodrug" refers to any pharmaceutically acceptable salt, ester, salt of an ester, or other derivative of a compound of the present invention, which, after administration to a subject, is capable of directly or indirectly providing a compound of the present invention or its pharmaceutically active metabolite or residue. Particularly preferred derivatives or prodrugs are compounds that, when administered to a patient, can enhance the bioavailability of the compounds of the present application (e.g., allow an oral compound to be more easily absorbed into the bloodstream) or contribute to the delivery of the parent compound to a biological organ or site of action (e.g., the brain or lymphatic system). Prodrugs can be prepared by modifying functional groups present in the compound in such a way that they are capable of breaking down to the parent compound by routine manipulation or in vivo. Various prodrug forms are well known in the art. See T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems (1987), Vol. 14 of the ACS Symposium Series; Bioreversible Carriers in Drug Design, (1987), Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press, also provides a discussion of prodrugs. Design of Prodrugs, Bundgaard, A. Ed., Elseview, 1985 and Method in Enzymology, Widder, K. et al., Ed.; Academic, 1985, vol. 42, pp. 309-396; Bundgaard, H. "Design and Application of Prodrugs" in A Textbook of Drug Design and Development, Krosgaard-Larsen and H. Bundgaard, Ed., 1991, Chapter 5, pp. 113-191; and Bundgaard, H., Advanced Drug Delivery Review, 1992, 8, 1-38, are incorporated herein by reference.

[0042] The term "stereoisomer" refers to an isomer resulting from differences in the spatial arrangement of atoms in a molecule. The compounds of the present invention contain structures such as asymmetric or chiral centers, double bonds, and the like. Therefore, the compounds of the present invention may include various isomers, such as optical isomers, geometric isomers, tautomers, and blocking isomers. These isomers, as well as their single isomers and racemates, are also within the scope of the present invention. For example, optical isomers can be prepared as optically active (R)- and (S)-isomers, as well as D- and L-isomers, by chiral resolution, chiral synthesis, chiral reagents, or other conventional techniques. For example, diastereoisomers can be separated by reaction with an appropriate optically active substance (e.g., chiral alcohol or Mosher's acid chloride) and converted into diastereoisomers, which can then be separated and converted (e.g., hydrolyzed) into the corresponding single isomers. Separation can also be achieved, for example, by chromatographic columns.

[0043] The "pharmaceutical compositions" herein can be prepared by methods well known in the pharmaceutical art and can be administered or administered by various routes. This depends on whether local or systemic treatment is required and the area to be treated. The "pharmaceutical compositions" herein can be administered topically (e.g., transdermally, via the skin, eye, and mucous membranes, including intranasal, vaginal, and rectal), pulmonary (e.g., inhalation or insufflation of powders or aerosols, including administration via a nebulizer; intratracheal, intranasal), orally, or parenterally. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration. Parenteral administration can be in the form of a single bolus dose or, for example, by a continuous infusion pump. Pharmaceutical compositions herein may be in the form of, but are not limited to, tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or dissolved in a liquid vehicle); ointments containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders, and the like.

[0044] The pharmaceutical compositions herein can be formulated in unit dosage form, with a single dosage containing about 0.1 to 1000 mg, usually about 5 to 1000 mg, and more usually about 100 to 500 mg of active ingredient. The term "unit dosage form" refers to physically discrete, single dosage units suitable for use in human patients and other mammals, each unit containing a predetermined amount of active material calculated to produce a desired therapeutic effect in admixture with a suitable pharmaceutical carrier.

[0045] The term "individual" refers to an individual, including mammals and non-mammals, suffering from a disease, condition, or pathology, etc. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); farm animals, such as cows, horses, sheep, goats, and pigs; domesticated animals, such as rabbits, dogs, and cats; and laboratory animals, including rodents, such as rats, mice, and guinea pigs.

[0046] The term "treatment" and other similar synonyms include ameliorating, alleviating, or improving symptoms of a disease or condition, preventing other symptoms, improving or preventing the underlying metabolic causes of a symptom, inhibiting a disease or condition, e.g., halting the progression of a disease or condition, ameliorating a disease or condition, improving a disease or condition, ameliorating symptoms caused by a disease or condition, or halting the symptoms of a disease or condition. Furthermore, the term may also encompass prophylactic purposes. The term also encompasses achieving a therapeutic and / or prophylactic effect. The therapeutic effect refers to curing or ameliorating the underlying disease being treated. Furthermore, curing or ameliorating one or more physiological symptoms associated with an underlying disease is also a therapeutic effect, e.g., when an improvement in the patient's condition is observed even though the patient is still affected by the underlying disease. For prophylactic effects, the composition or compound can be administered to a patient at risk of developing a particular disease, or to a patient who is experiencing one or more physiological symptoms of the disease, even if the disease has not yet been diagnosed.

[0047] The term "amount required to achieve the desired therapeutic effect" or "therapeutically effective amount" refers to an amount of at least one drug or compound sufficient, upon administration, to ameliorate to some extent one or more symptoms of the disease or condition being treated. The result may be reduction and / or amelioration of the signs, symptoms, or etiology of the disease, or any other desired change in a biological system. Techniques such as dose escalation studies can be used to determine the appropriate effective amount for any individual case. The actual dosage of a compound, pharmaceutical composition, or drug will typically be determined by a physician based on relevant circumstances, such as the condition being treated, the selected route of administration, the compound actually administered, the age, weight, and response of the individual patient, and the severity of the patient's symptoms.

[0048] The proportion or concentration of a compound of the present invention in a pharmaceutical composition does not need to be fixed and will depend on various factors, such as dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, for parenteral administration, the compound of the present invention may be provided in an aqueous physiological buffer solution containing about 0.1 to 10% w / v of the compound. A typical dosage range is about 1 μg / kg to about 1 g / kg body weight / day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg body weight / day. Dosage will likely depend on variables such as the type and progression of the disease or condition, the general health of the particular patient, the relative biological potency of the selected compound, the excipient formulation, and its route of administration.

[0049] The term "administration" refers to a method that allows a compound or composition to be delivered to a desired site of biological action. These methods include, but are not limited to, oral, intraduodenal, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, and intraarterial injection or infusion), topical, and rectal administration. Those skilled in the art are familiar with administration techniques that can be used for the compounds and methods described herein. For example, see Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.

[0050] The term “IC 50 " means a 50% inhibition of the maximal effect obtained in the assay measuring such effect.

[0051] compound Meanwhile, the present application provides an azacycloalkane compound represented by formula I or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled form, isomer, or prodrug thereof: JPEG2026504901000003.jpg155154 where R1 is -NHR 11 , -OR 11 , -SR 11 or -C(=O)NHR 11 where R 11 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~6 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, substituted or unsubstituted C 5~20 Heteroaryl, substituted or unsubstituted C1~6 Alkyl acyl, substituted or unsubstituted C 1~6 Alkyl sulfonyl, substituted or unsubstituted C 5~20 Heteroarylacyl, and substituted or unsubstituted C 1~6 alkoxy; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , -C(=O)R 33 , hydroxyl, sulfhydryl, substituted C 3~8 Cycloalkyl, substituted C 3~8 Heterocyclyl, substituted C 6~20 Aryl and substituted C 5~20 selected from the group consisting of heteroaryl; R2 is hydrogen, halogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 1~6 Haloalkyl, substituted or unsubstituted C 1~6 Alkoxy, cyano, -NR 31 R 32 , hydroxyl, carboxy, and sulfhydryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R3 is selected from the group consisting of cyano, -CONH2 and carboxy; L is -S(=O)2-, -C(=O)-, -CH2-, and -S(=O)(=N)R L -, where R L is hydrogen, and substituted or unsubstituted C 1~8 alkyl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R4 is halogen, substituted or unsubstituted C 1~8 Alkyl, -NR 31 R 32 , substituted or unsubstituted C 1~8 Alkoxy, substituted or unsubstituted C 2-8 Alkenyl, substituted or unsubstituted C 2-8 Alkynyl, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, or substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, hydrazide group, C 1~8 Alkylsulfonyl, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, -COR 33 , selected from the group consisting of carboxy and sulfhydryl; A1 is selected from C atoms; W1, W2, W3, W4 and W5 are each independently a C atom or an N atom, and one, two or three of W1, W2, W3, W4 and W5 are N atoms; a dotted circle represents a bond that forms an aromatic ring; R 21 is hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 22 is hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 23 is hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 24 is hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 25 is hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; Alternatively, R 24 , R 24 W4 to which is bonded, and R 25 , and R 25 are joined together to form a 5-membered aryl, cycloalkyl, heteroaryl, or heterocycloalkylcyclyl fused to a 6-membered ring consisting of A, W, W, W, W, and W; R 31 and R 32 are each independently hydrogen, sulfonamido, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, hydroxyl, -C(=O)OR 34 and sulfhydryl; R 33 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Alkoxy, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 34 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Haloalkyl, substituted or unsubstituted C 1~8 Alkoxy, and substituted or unsubstituted C 3~8 cycloalkyl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy, and sulfhydryl; n is 1, 2 or 3; wherein the compound is not: JPEG2026504901000004.jpg245161JPEG2026504901000005.jpg84159The inventors have found through their research that the compounds of the present invention have good Rho kinase inhibitory activity, can be used as inhibitors targeting ROCK, and can be used to regulate diseases mediated by Rho kinase.

[0052] In one embodiment, R1 is -NHR 11 where R 11 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~6 Cycloalkyl, substituted or unsubstituted C 3~8Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, substituted or unsubstituted C 5~20 Heteroaryl, substituted or unsubstituted C 1~6 Alkyl acyl, substituted or unsubstituted C 1~6 Alkyl sulfonyl, substituted or unsubstituted C 5~20 Heteroarylacyl, and substituted or unsubstituted C 1~6 alkoxy; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , -C(=O)R 33 , hydroxyl, sulfhydryl, substituted C 3~8 Cycloalkyl, substituted C 3~8 Heterocyclyl, substituted C 6~20 Aryl and substituted C 5~20 selected from the group consisting of heteroaryl; where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20Heteroaryl, cyano, hydroxyl, -C(=O)OR 34 and sulfhydryl; R 33 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Alkoxy, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 34 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Haloalkyl, substituted or unsubstituted C 1~8 Alkoxy, and substituted or unsubstituted C 3~8 cycloalkyl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 It is selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy and sulfhydryl.

[0053] In one embodiment, R1 is -NHR 11 where R 11 is hydrogen and substituted or unsubstituted C 1~6 alkyl, and the substituents are selected from the group consisting of -NR31 R 32 , -C(=O)R 33 , and substituted or unsubstituted C 3~8 heterocyclyl. For example, R 11 can be selected from the group consisting of hydrogen, optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted isopropyl, optionally substituted butyl, optionally substituted tert-butyl, optionally substituted pentyl, and optionally substituted hexyl, and the substituents are selected from the group consisting of -NR 31 R 32 , -C(=O)R 33 , and substituted or unsubstituted C 3~8 Heterocyclyl (substituents are C 1~6 alkyl, such as methyl, ethyl, propyl, isopropyl, etc. Preferably, R 11 is a substituted or unsubstituted C 1~6 alkyl, and the substituents are selected from the group consisting of -NR 31 R 32 where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 3~8 Heterocyclyl (substituents are C 1~6 alkyl, such as methyl, ethyl, propyl, isopropyl, etc. Preferably, R 11 is a substituted or unsubstituted C 1~6 alkyl, and the substituent is selected from the group consisting of -C(=O)R 33 where R 33 are independently substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 3~8 Heterocyclyl (substituents are C 1~6 alkyl, which may be, for example, methyl, ethyl, propyl, isopropyl, etc.

[0054] JPEG2026504901000006.jpg69165 In one embodiment, R1 is -OR 11 where R 11 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~6 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, substituted or unsubstituted C 5~20 Heteroaryl, substituted or unsubstituted C 1~6 Alkyl acyl, substituted or unsubstituted C 1~6 Alkyl sulfonyl, substituted or unsubstituted C 5~20 Heteroarylacyl, and substituted or unsubstituted C 1~6 alkoxy; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , -C(=O)R 33 , selected from the group consisting of hydroxyl and sulfhydryl; Preferably, R 11 is hydrogen, and substituted or unsubstituted C 1~6 alkyl, and the substituents are selected from the group consisting of -NR 31 R 32 , -C(=O)R 33 , and substituted or unsubstituted C 3~8 heterocyclyl; where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, hydroxyl, -C(=O)OR 34 and sulfhydryl; R 33 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Alkoxy, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 34 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Haloalkyl, substituted or unsubstituted C 1~8 Alkoxy, and substituted or unsubstituted C 3~8 cycloalkyl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 It is selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy and sulfhydryl.

[0055] In one embodiment, R1 is -OR 11 where R 11 is hydrogen, or C 1~6 Alkyl (e.g., C such as methyl, ethyl, propyl, and isopropyl) 1~3 alkyl).

[0056] In one embodiment, R1 is -C(=O)NHR 11 where R 11 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~6 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, substituted or unsubstituted C 5~20 Heteroaryl, substituted or unsubstituted C 1~6 Alkyl acyl, substituted or unsubstituted C 1~6 Alkyl sulfonyl, substituted or unsubstituted C 5~20 Heteroarylacyl, and substituted or unsubstituted C 1~6 alkoxy; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , -C(=O)R 33, selected from the group consisting of hydroxyl and sulfhydryl; Preferably, R 11 is hydrogen, and substituted or unsubstituted C 1~6 alkyl, and the substituents are selected from the group consisting of -NR 31 R 32 , -C(=O)R 33 , and substituted or unsubstituted C 3~8 heterocyclyl; where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 2~6 Alkenyl, substituted or unsubstituted C 2~6 Alkynyl, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, hydroxyl, -C(=O)OR 34 and sulfhydryl; R 33 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Alkoxy, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , selected from the group consisting of hydroxyl, carboxy, and sulfhydryl; R 34 is hydrogen, substituted or unsubstituted C 1~8 Alkyl, substituted or unsubstituted C 1~8 Haloalkyl, substituted or unsubstituted C 1~8 Alkoxy, and substituted or unsubstituted C 3~8 cycloalkyl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 It is selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy and sulfhydryl.

[0057] In one embodiment, R1 is -C(=O)NHR 11 where R 11 is hydrogen, or C 1~6 Alkyl (e.g., C such as methyl, ethyl, propyl, and isopropyl) 1~3 alkyl).

[0058] In each of the above embodiments relating to R, R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, or substituted or unsubstituted C 3~8 Heterocyclyl (wherein the substituents are C substituted, e.g., methyl, ethyl, propyl, isopropyl, etc.) 1~6 R may be alkyl. 33 are independently substituted or unsubstituted C 1~6 Alkyl, or substituted or unsubstituted C 3~8Heterocyclyl (wherein the substituents are C substituted, e.g., methyl, ethyl, propyl, isopropyl, etc.) 1~6 It can be alkyl.

[0059] In one embodiment, R2 is selected from hydrogen and R3 is selected from cyano.

[0060] The dotted circle in Formula I indicates a bond that forms an aromatic ring. In one embodiment, W1, W2, W3, W4, and W5 are each independently selected from the group consisting of a C atom and an N atom, and one, two, or three of W1, W2, W3, W4, and W5 are N atoms. In one embodiment, two of W1, W2, W3, W4, and W5 are N atoms. In one embodiment, three of W1, W2, W3, W4, and W5 are N atoms. In one embodiment, W3 is an N atom, and one or two of W1, W2, W4, and W5 are N atoms.

[0061] In one aspect, JPEG2026504901000007.jpg83159In the structures A1 to A7, R7 is independently hydrogen, halogen, or -NR 31 R 32 , and substituted or unsubstituted C 5~20 heteroaryl; where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 selected from the group consisting of heteroaryl; where the substituents are halogen, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20It is selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy and sulfhydryl.

[0062] In the above structural formula, * indicates the position of connection to the pyrazole ring structure.

[0063] In one embodiment, each R7 is independently hydrogen, -NR 31 R 32 , and substituted or unsubstituted C 5~20 heteroaryl; where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 5~20 Heteroaryl, and substituted or unsubstituted C 3~8 heterocyclyl; where the substituent is C 1~8 is selected from alkyl.

[0064] JPEG2026504901000008.jpg140165Here, in the A2 structure, R7 is -NR 31 R 32 , or substituted or unsubstituted C 5~20 is heteroaryl, where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 5~20 selected from the group consisting of heteroaryl; where the substituent is C 1~8 It is selected from the group consisting of alkyl and hydroxyl.

[0065] JPEG2026504901000009.jpg60156Here, in the A3 structure, R7 is -NR 31 R 32 , and substituted or unsubstituted C 5~20 is heteroaryl, where R 31 and R 32are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 5~20 selected from the group consisting of heteroaryl; where the substituent is C 1~8 It is selected from the group consisting of alkyl and hydroxyl.

[0066] JPEG2026504901000010.jpg66161Here, in the A4 structure, R7 is -NR 31 R 32 , or substituted or unsubstituted C 5~20 is heteroaryl, where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 5~20 selected from the group consisting of heteroaryl; where the substituent is C 1~8 It is selected from the group consisting of alkyl and hydroxyl.

[0067] JPEG2026504901000011.jpg62154Here, in the A5 structure, R7 is -NR 31 R 32 , or substituted or unsubstituted C 5~20 is heteroaryl, where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 5~20 selected from the group consisting of heteroaryl; where the substituent is C 1~8 It is selected from the group consisting of alkyl and hydroxyl.

[0068] JPEG2026504901000012.jpg65150Here, in the A6 structure, R7 is -NR 31 R 32 , or substituted or unsubstituted C 5~20 is heteroaryl, where R31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 5~20 selected from the group consisting of heteroaryl; where the substituent is C 1~8 It is selected from the group consisting of alkyl and hydroxyl.

[0069] JPEG2026504901000013.jpg66144Here, in the A7 structure, R7 is -NR 31 R 32 , or substituted or unsubstituted C 5~20 is heteroaryl, where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1~6 Alkyl, and substituted or unsubstituted C 5~20 selected from the group consisting of heteroaryl; where the substituent is C 1~8 It is selected from the group consisting of alkyl and hydroxyl.

[0070] In the above embodiments, R7 is -NHCH3, -NHCH2CH2OH, Cl, It could also be JPEG2026504901000014.jpg49164.

[0071] In one embodiment, L is -S(=O)2-, -C(=O)- and -S(=O)(=N)R L -, where R L is hydrogen and C 1~6 In one embodiment, L is -C(=O)-. In one embodiment, L is -S(=O)(=N)R. L - where R L is hydrogen or a C alkyl group such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, pentyl, and hexyl. 1~6alkyl.

[0072] In one embodiment, R4 is a substituted or unsubstituted C 1~8 Alkyl, -NR 31 R 32 , substituted or unsubstituted C 1~8 Alkoxy, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, hydrazide group, C 1~8 Alkylsulfonyl, C 1~8 Alkyl, C 1~8 Haloalkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, C 3~8 Heterocyclyl, C 6~20 Aryl, C 5~20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, -COR 33 , selected from the group consisting of carboxy and sulfhydryl; where R 31 and R 32 are each independently hydrogen, sulfonamido, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, cyano, -NR 31 R 32 , hydroxyl, -C(=O)OR 34 and sulfhydryls.

[0073] In one embodiment, R4 is a substituted or unsubstituted C 1~8 alkyl (e.g., methyl, ethyl, propyl, etc.), where the substituents are halogen, hydroxyl, cyano, C 1~8 It may be alkylsulfonyl, etc., for example, R4 may be methyl, ethyl, trifluoroethyl, trifluoromethyl, methylsulfonylmethyl, etc.

[0074] In one embodiment, R4 is a substituted or unsubstituted C 3~8 and selected from the group consisting of cycloalkyl (e.g., cyclopropyl, cyclobutyl, etc.), where the substituents may be halogen, hydroxyl, cyano, etc., e.g., R4 may be 2,2-difluorocyclopropyl, 2-cyanocyclopropyl, etc.

[0075] In one embodiment, R4 is -NR 31 R 32 where R 31 and R 32 are each independently hydrogen, sulfonamido, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 1~6 Alkyl, substituted or unsubstituted C 3~8 Cycloalkyl, substituted or unsubstituted C 3~8 Heterocyclyl, substituted or unsubstituted C 6~20 Aryl, and substituted or unsubstituted C 5~20 heteroaryl; the substituents are selected from the group consisting of halogen, C 1~8 Alkyl, C 1~8 Alkoxy, C 3~8 Cycloalkyl, cyano, -NR 31 R 32 , hydroxyl, -C(=O)OR 33 and sulfhydryl; R 33 is hydrogen and C 1~6 In one embodiment, R4 is selected from the group consisting of -NR 31 R 32 where R 31 is hydrogen and R32 is a substituted or unsubstituted C 1~8 Alkyl (e.g., methyl, ethyl, propyl, etc.), substituted or unsubstituted C 3~8 Cycloalkyl (e.g., cyclopropyl), and substituted or unsubstituted C 5~20 In one embodiment, R4 is selected from the group consisting of -NR 31 R 32 where R 31 is hydrogen and R 32 are independently substituted or unsubstituted C 1~6 alkyl (e.g., methyl, ethyl, propyl, etc.), and the substituent is -C(=O)OR 34 where R 34 is hydrogen or C 1~6 It is selected from the group consisting of alkyl (e.g., methyl, ethyl, propyl, etc.).

[0076] In one embodiment, n is 1, 2 or 3, in particular 1 or 2. When n is 1, the azacycloalkane ring is azetidinyl; when n is 2, the azacycloalkane ring is azacyclopentyl; and when n is 3, the azacycloalkane ring is azacyclohexyl.

[0077] In one embodiment, the azacycloalkane compound is selected from the group consisting of the following compounds: JPEG2026504901000015.jpg211170JPEG2026504901000016.jpg245170JPEG2026504901000017.jpg230170JPEG2026504901000018.jpg219170JPEG2026504901000019.jpg184166In a preferred embodiment, the azacycloalkane compound is selected from the group consisting of the following compounds: JPEG2026504901000020.jpg217170JPEG2026504901000021.jpg110170In the present invention, the compounds described in the present invention can be prepared by the following methods. The following methods and examples are intended to illustrate these methods. These processes and examples should not be construed as limiting the present invention in any way. The compounds described herein can be synthesized by standard synthetic techniques known to those skilled in the art, or can be synthesized by combining methods known in the art with the methods described herein.

[0078] The chemical reactions of the examples of the present invention are carried out in suitable solvents, which must be appropriate for the chemical transformations of the present invention and the reagents and materials required therefor. To obtain the compounds of the present invention, it may be necessary for those skilled in the art to modify or select synthetic steps or reaction processes based on existing embodiments.

[0079] An important consideration in planning any synthetic route in this field is the selection of an appropriate protecting group for the reactive functional group (e.g., amino in the present invention). For the trained practitioner, Greene and Wuts (Protective Groups In Organic Synthesis, Wiley and Sons, 1991) is an authority in this area. All references cited herein are incorporated herein in their entirety.

[0080] The reactions described herein can be monitored according to any suitable method known in the art. For example, product formation can be monitored by optical methods such as nuclear magnetic resonance spectroscopy (e.g., H or C), infrared spectroscopy, spectrophotometry (e.g., UV-visible light), mass spectrometry, or chromatography, such as high performance liquid chromatography (HPLC) or thin layer chromatography.

[0081] The compounds of general formula I of the present invention can be prepared by one skilled in the art of organic synthesis using standard methods in the art according to the following process.

[0082] Process 1: JPEG2026504901000022.jpg47166 Compound a1 undergoes a Michael addition reaction with compound a2 to produce compound a3, compound a3 undergoes removal of the protecting group to give compound a4, and compound a4 undergoes a substitution reaction with compound a5 to give the compound of formula I of the present invention.

[0083] Process 2: JPEG2026504901000023.jpg68143 Compound b1 undergoes a coupling reaction with compound b2 to produce compound b3, compound b3 reacts with a substituted boronic acid or a substituted amino compound to produce compound b4, compound b4 undergoes removal of the protecting group to produce compound b5, and compound b5 undergoes a Michael addition reaction with compound b6 to produce the compound of formula I of the present invention.

[0084] Pharmaceutical Compositions and Uses The present invention also provides a pharmaceutical composition comprising any one of the compounds of the above technical configurations, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, and a pharmaceutically acceptable carrier.

[0085] The pharmaceutical composition may be in the form of, but not limited to, oral dosage forms, parenteral dosage forms, topical dosage forms, rectal dosage forms, etc. For example, the pharmaceutical composition may be oral tablets, capsules, pills, powders, sustained-release formulations, solutions and suspensions, sterile solutions, suspensions or emulsions for parenteral gastrointestinal injection, ointments, creams, gels, etc. for external use, eye drops for external use, inhalants for external use, or suppositories for rectal administration.

[0086] The pharmaceutical composition may also contain other active ingredients or drugs, which are used in combination (drug combination) with the compound or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled form, isomer, or prodrug thereof.

[0087] The present invention also provides use of the above-mentioned compound or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled product, isomer, or prodrug thereof, as well as the above-mentioned pharmaceutical composition, in the preparation of a medicament for treating a disease mediated by Rho kinase, wherein Rho kinase includes types such as ROCK1 and ROCK2.

[0088] The present application also relates to a method for treating a disease mediated by Rho kinase, comprising administering to a patient in need thereof a therapeutically effective amount of the above-described azacycloalkane compound or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, or the above-described pharmaceutical composition, wherein Rho kinase includes types such as ROCK1 and ROCK2.

[0089] In one embodiment, the disease mediated by Rho kinase is asthma, cancer, glaucoma, insulin resistance, renal failure, neurodegeneration, or osteoporosis.

[0090] The azacycloalkane compounds and pharmaceutical compositions thereof provided by the present invention have significant Rho kinase inhibitory activity, and their enzymatic and cellular activities are superior to those of existing ROCK inhibitors such as ripasudil, netarsudil, and vermosudil, and have great potential for application.

[0091] To make the objectives, technical configurations and advantages of the present invention clearer, the technical configurations of exemplary embodiments of the present invention are further described below.

[0092] Example 1: 2-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)acetonitrile JPEG2026504901000024.jpg75106Synthetic route: JPEG2026504901000025.jpg66170 Step A: tert-Butyl 3-(cyanomethyl)-3-(3-(1,3-dioxoisoindol-2-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate JPEG2026504901000026.jpg8112746g (0.1 mol, 1.0 eq) 2-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-3-yl)isoindole-1,3-dione and 19.5 g (0.1 mol, 1.0 eq) tert-butyl 3-(cyanomethyl)azetidine-1-carboxylate were dissolved in 200mL N,N-dimethylformamide, and 1.5 g (0.01 mol, 0.1 eq) 1,8-diazabicyclo[5.4.0]undec-7-ene was slowly added dropwise to the reaction mixture at room temperature, followed by stirring overnight. The reaction was quenched with water, the aqueous phase was extracted with ethyl acetate, the aqueous phase was washed twice with water, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the product (61 g, yield=93%).

[0093] Step B: 2-(3-(3-(1,3-dioxoisoindol-2-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile hydrochloride JPEG2026504901000027.jpg6811861 g (93 mmol, 1.0 eq) tert-butyl 3-(cyanomethyl)-3-(3-(1,3-dioxoisoindol-2-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate was dissolved in 300 mL dichloromethane and 50 mL 4N hydrochloric acid in dioxane was added at room temperature. The reaction was stirred overnight at room temperature. TLC monitored the completion of the reaction. The reaction was evaporated to dryness to give the product (55 g, yield = 100%).

[0094] Step C: 2-(3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)acetonitrile 2-(3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (3 g, 5.41 mmol, 1.0 eq) was dissolved in dichloromethane (30 mL) in an ice bath. Triethylamine (3.2 g, 32.46 mmol, 6.0 eq) was slowly added dropwise to the reaction mixture and stirred for 15 min. Next, 2,2,2-trifluoroethane-1-sulfonyl chloride (987 mg, 5.41 mmol, 1.0 eq) dissolved in 5 mL of dichloromethane was added dropwise to the reaction mixture and reacted at 25 °C for 2 h. After completion of the reaction, the reaction mixture was evaporated under reduced pressure to dryness. The residue obtained by concentrating the reaction mixture was purified by column chromatography (PE:EA=1:2) to obtain the product, 2-(3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(2-trimethylsilylethoxy)methyl)-7H-pyrrolopyrrolidin-4-yl-1H-pyrazoline-1-(2,2,2-trifluoroethylsulfonyl)azetidin-3-yl)acetonitrile (2.1 g, 55.41%), as a yellow oil.

[0095] LC-MS (ESI), m / z: [M+H] + =700.9.

[0096] Step D: 2-(3-(3-(1,3-dioxoisoindol-2-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)acetonitrile JPEG2026504901000029.jpg831212-(3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)acetonitrile (700 mg, 1.0 mmol, 1.0 eq) was dissolved in acetonitrile (6 mL). Boron trifluoride-ethyl ether solution (2 mL) was added dropwise in an ice bath. The reaction mixture was stirred at 25°C for 2 h. After completion of the reaction, the mixture was evaporated to dryness under reduced pressure. The crude product, 2-(3-amino-4-methylol-7H-pyrrolyl)pyrimidin-4-yl-1-pyrazole-1-(2,2,2-trifluoroethylsulfonyl)azetidin-3-ylacetonitrile (450 mg, crude) was obtained as a yellow oil.

[0097] LC-MS (ESI), m / z: [M+H] + =601.

[0098] Step E: 2-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)acetonitrile JPEG2026504901000030.jpg761152-(3-(3-(1,3-dioxoisoindol-2-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)acetonitrile (450 mg, 0.75 mmol) was dissolved in methanol (7 mL) and ethylenediamine (1 mL) was added in an ice bath. After the reaction was completed, water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL*3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The residue was purified by Prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-HO (0.1% FA), gradient 20%-50%, flow rate: 20 mL / min) to obtain the product (45 mg, yield = 11%).

[0099] LC-MS (ESI), m / z: [M+H] + =440.9.

[0100] 1 H-NMR(400MHz, D2O) δ 8.61 (s, 1H), 8.29 (s, 1H), 7.51 (d, J = 3.6 Hz, 1H), 6.76 (d, J = 3.6 Hz, 1H), 4.33 (d, J = 9.3 Hz, 2H), 3.88 (s, 4H), 3.44 (s, 2H). The following examples were prepared with reference to the experimental routes and methods of Example 1: JPEG2026504901000031.jpg254166JPEG2026504901000032.jpg253165JPEG2026504901000033.jpg252165JPEG2026504901000034.jpg132164Example 10: 3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)-3-(cyanomethyl)-N-methylazetidine-1-formamide JPEG2026504901000035.jpg75101Synthetic route: JPEG2026504901000036.jpg89153 Step A: 2-{3-[3-(1,3-dioxoisoindole)-4-[7-(methylol)pyrrolidine[2,3-d]pyrimidine]pyrazole]3-azetidine}acetonitrile JPEG2026504901000037.jpg69112 tert-Butyl 3-(cyanomethyl)-3-[3-(1,3-dioxoisoindol-2-yl)-4-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]azetidine-1-carboxylate (10.1 g, 0.015 mol) was added to dichloromethane (200 mL) at room temperature and completely dissolved. After this, trifluoroacetic acid (100 mL) was added and the mixture was stirred overnight in an oil bath at 30°C. Rotary evaporation to dryness gave the product 2-{3-[3-(1,3-dioxoisoindole)-4-[7-(methylol)pyrrolidine[2,3-d]pyrimidine]pyrazole]3-azetidine}acetonitrile (12.0 g, crude).

[0101] LC-MS (ESI), m / z: [M+H] + = 455.0.

[0102] Step B: 2-[3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)azetidin-3-yl]acetonitrile JPEG2026504901000038.jpg65872-{3-[3-(1,3-dioxoisoindole)-4-[7-(methylol)pyrrolidine[2,3-d]pyrimidine]pyrazole]3-azetidine}acetonitrile (12 g, 0.026 mol) was added to methanol (100 mL) at room temperature. After complete dissolution, ethylenediamine (4.68 g, 0.078 mol) was added and the mixture was allowed to react at room temperature for 4 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL * 5). The organic phase was collected, dried, and concentrated under reduced pressure. The residue was purified on a C18 column to give the product 2-[3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)azetidin-3-yl]acetonitrile (1.9 g, 25.1%).

[0103] LC-MS (ESI), m / z: [M+H] + = 295.0.

[0104] Step C: Phenyl N-methylcarbamate To a solution of methylamine (2 g, 0.03 mol) in tetrahydrofuranol (15 mL) was added sodium bicarbonate (2.77 g, 0.03 mol) at 25°C. The reaction mixture was stirred at 25°C for 15 minutes. Phenylchloroformate (4.5 g, 0.03 mol) was added, and the reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched with water (30 mL) and extracted with ethyl acetate (20 mL*3). The organic phase was dried and concentrated. The residue obtained by concentrating the reaction mixture was purified by column chromatography (DCM:MeOH=20:1) to give the product (1 g, 22%).

[0105] LC-MS (ESI), m / z: [M+H] + = 152.1.

[0106] Step D: 3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)-3-(cyanomethyl)-N-methylazetidine-1-formamide JPEG2026504901000040.jpg7498 At room temperature, phenyl N-methylcarbamate (51 mg, 0.339 mmol) and 2-[3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)azetidin-3-yl]acetonitrile (99.77 mg, 0.339 mmol) were added to N,N-dimethylformamide (2.5 mL) and stirred for 5 minutes. N,N-Diisopropylethylamine (131 mg, 1.017 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated, and the residue was purified by prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% NH3H2O), gradient 25%-30%, flow rate: 20 mL / min) to obtain the product (20 mg, 16.72%).

[0107] LC-MS (ESI), m / z: [M+H] + = 352.0.

[0108] 1 H-NMR(400MHz, DMSO-d6) δ 12.07 (s, 1H), 8.66 (s, 1H), 8.57 (s, 1H), 7.59 - 7.52 (m, 1H), 7.11 - 7.05 (m, 1H), 6.50 (d, J = 4.5 Hz, 1H), 6.38 (s, 2H), 4.38 (d, J = 9.0 Hz, 2H), 4.03 (d, J = 8.9 Hz, 2H), 3.58 (s, 2H), 2.57 (d, J = 4.5 Hz, 3H).

[0109] The following examples were prepared with reference to the experimental route and methods of Example 10: JPEG2026504901000041.jpg253157JPEG2026504901000042.jpg255153JPEG2026504901000043.jpg238157JPEG2026504901000044.jpg56158Example 21: 3-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidin-1-yl]-3-oxopropionitrile JPEG2026504901000045.jpg76116Synthetic route: JPEG2026504901000046.jpg50158 Step A: 3-(3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-1-yl)-3-oxopropionitrile JPEG2026504901000047.jpg771362-Cyanoacetic acid (92 mg, 1.08 mmol, 2.0 eq), triethylamine (220 mg, 2.2 mmol, 4.0 eq), and dichloromethane (5 mL) were mixed with 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (410.4 mg, 1.08 mmol, 2.0 eq). The reaction mixture was heated at 25°C for 0.5 min. The reaction mixture was stirred for 2 h, and then 2-(3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4))-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (300 mg, 0.54 mmol, 1.0 eq) was added and the reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with water (20 mL) and extracted with dichloromethane (20 mL*3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The crude product was passed through a Combiflash column (dichloromethane:methanol=35:1) to give the product 3-(3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl))-1H-pyrazol-1-yl)azetidin-1-yl)-3-oxopropionitrile (220 mg, 65% yield) as a white solid.

[0110] LC-MS(ESI), m / z: [M+H] + =622.0.

[0111] Step B: 3-(3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl))-1H-pyrazol-1-yl)azetidin-1-yl)-3-oxopropionitrile 3-(3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-1-yl)-3-oxopropionitrile (220 mg, 0.35 mmol, 1.0 eq) was added to dichloromethane (5 mL) at room temperature, and boron trifluoride ethyl ether solution (0.5 mL) was added at 0 °C. The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (10 mL*3), the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the crude product (140 mg, yield 76%).

[0112] LC-MS(ESI), m / z: [M+H] + =522.0.

[0113] Step C: 3-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidin-1-yl)-3-oxopropionitrile 3-(3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl))-1H-pyrazol-1-yl)azetidin-1-yl)-3-oxopropionitrile (140 mg, 0.27 mmol, 1.0 eq) was dissolved in methanol (5 mL) at room temperature, and ethylenediamine (0.5 mL) was added in an ice bath to adjust the pH to 8–9. The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated and purified by Prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-HO (0.1% TFA), gradient 25%-40%, flow rate: 20 mL / min) to obtain the product (31.9 mg, 33%).

[0114] LC-MS(ESI), m / z: [M+H] + =362.0.

[0115] 1 H-NMR(400MHz, DMSO-d6) δ 8.63 (d, J = 9.9 Hz, 1H), 8.57 (d, J = 7.4 Hz, 1H), 7.54 (d, J = 5.3 Hz, 1H), 7.03 (d, J = 9.7 Hz, 1H), 4.67 (d, J = 12.9 Hz, 1H), 4.52 (d, J = 10.6 Hz, 1H), 4.38 (d, J = 10.6 Hz, 1H), 4.14 (d, J = 12.1 Hz, 1H), 3.78 (s, 2H), 3.56 (d, J = 9.3 Hz, 2H).

[0116] The following examples were prepared with reference to the experimental route and methods of Example 21: JPEG2026504901000050.jpg253161JPEG2026504901000051.jpg254162JPEG2026504901000052.jpg225161JPEG2026504901000053.jpg255158JPEG2026504901000054.jpg146161Example 36: 2-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-(S-methylsulfonylimino)azetidin-3-yl)acetonitrile JPEG2026504901000055.jpg7497Synthetic route: JPEG2026504901000056.jpg61156 Step A: N-(tert-butyldimethylsilyl)methanesulfonamide Under nitrogen gas protection, methylsulfonamide (1 g, 1.53 mmol, 1.0 eq) was dissolved in tetrahydrofuran (15 mL), triethylamine (464 mg, 4.59 mmol, 3.0 eq) was added, and a solution of tert-butyldimethylchlorosilane (462 mg, 3.06 mmol, 2.0 eq) in tolyl (50 mL) was added dropwise. The mixture was allowed to react overnight at room temperature, filtered, and the solid was washed with ethyl ether (200 mL). The filtrate was added with ethyl ether (20 mL), allowed to stand for 30 min, filtered again, and concentrated to give the crude product. The crude product was purified through a Combiflash column (petroleum ether:ethyl acetate = 1:1) to give the product N-(tert-butyldimethylsilyl)methanesulfonamide (1.2 g, 39% yield) as a white solid.

[0117] LC-MS(ESI), m / z: [M+H] + =210.0.

[0118] Steps B & C: 2-(1-N-(tert-butyldimethylsilyl)-S-methylsulfonylimino)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy))methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile JPEG2026504901000058.jpg88157Under nitrogen gas protection, a 0.36 M dichlorotriphenylphosphine / chloroform suspension (15 mL, 5.4 mmol, 6.0 eq) was cooled to 0°C, triethylamine (810 mg, 8.1 mmol, 9.0 eq) was added, and the mixture was stirred for 15 min. After that, N-(tert-butyldimethylsilyl)methanesulfonamide (1.13 g, 5.4 mmol, 6.0 eq) was added at 0°C, and the mixture was stirred for 20 min. After that, the reaction mixture was cooled to 0°C and 2-(3-(3-(1,3-dioxoisoindolin-2-yl))-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4))-yl)-1-hydro-pyrazol-1-yl)azetidin-3-yl)acetonitrile (500 The mixture was added dropwise to a solution of 1 mg (0.90 mmol, 1.0 eq) of methyl methylpropional in dichloromethane (2 mL). After the addition, the mixture was warmed to room temperature and stirred for 2 h. The reaction was quenched by adding water and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The crude product was purified by passing through a Combiflash column (dichloromethane:methanol = 97:3) to give the crude product (1.1 g, purity: 50%, yield: 82%) as a yellow solid.

[0119] LC-MS(ESI), m / z: [M+H] + =746.0.

[0120] Step D: 2-(3-(3-(1,3-dioxoisoindolin-2-yl))-4-(7-(methylol)-7hydro-pyrrolo[2,3-d]pyrimidin-4-yl)-1-hydro-pyrazol-1-yl)-1-(S-methylsulfonylimino)azetidin-3-yl)acetonitrile 2-(1-N-(tert-butyldimethylsilyl)-S-methylsulfonylimino)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy))methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (1.1 g, 1.34 mmol, 1.0 eq) was added to dichloromethane (10 mL) at room temperature, and boron trifluoride ethyl ether solution (1 mL) was added at 0 °C. The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the crude product (500 mg, crude product).

[0121] LC-MS(ESI), m / z: [M+H] + =531.9.

[0122] Step E: 2-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-(S-methylsulfonylimino)azetidin-3-yl)acetonitrile 2-(3-(3-(1,3-dioxoisoindolin-2-yl))-4-(7-(methylol)-7-hydro-pyrrolo[2,3-d]pyrimidin-4-yl)-1-hydro-pyrazol-1-yl)-1-(S-methylsulfonylimino)azetidin-3-yl)acetonitrile (500 mg, crude) was dissolved in methanol (10 mL) at room temperature, and ethylenediamine (0.5 mL) was added in an ice bath to adjust the pH to 8–9. The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated and purified by Prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-HO (0.1% TFA), gradient 30%-35%, flow rate: 20 mL / min) to obtain the product (76 mg, 26%).

[0123] LC-MS(ESI), m / z: [M+H] + =372.0.

[0124] 1 H-NMR(400 MHz, DMSO-d6) δ 11.96 (d, 1H), 8.66 (s, 1H), 8.62 (s, 1H), 7.56 (d, J = 3.6 Hz, 1H), 7.12 (d, J = 3.6 Hz, 1H), 6.40 (s, 2H), 4.43 (dd, J = 11.7, 9.3 Hz, 2H), 4.03 (d, J = 9.6 Hz, 2H), 3.92 (s, 1H), 3.53 (s, 2H), 2.96 (s, 3H).

[0125] The following examples were prepared with reference to the experimental route and methods of Example 36: JPEG2026504901000061.jpg230155 Example 40: 3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)-N-(2,2,2-trifluoroethyl)azetidine-1-sulfonamide JPEG2026504901000062.jpg8597Synthetic route: JPEG2026504901000063.jpg74164 Step A: tert-Butyl (chlorosulfonyl)carbamate To a solution of tert-butyl alcohol (2.73 g, 0.037 mol, 1.3 eq) in dichloromethane (20 mL) was added [(chlorosulfonyl)imino]methanone (4 g, 0.028 mol, 1.0 eq) at 30 °C under nitrogen gas protection, and the resulting reaction mixture was stirred at 25 °C for 20 min. After the reaction was complete, the mixture was concentrated to half its original volume and placed in a refrigerator under nitrogen gas protection for 50 min. n-Hexane (10 mL) was added, and the reaction mixture was filtered. The precipitate was removed by filtration with n-hexane. The filter cake was collected and dried to give the product, tert-butyl (chlorosulfonyl)carbamate (2 g, crude), which was used directly in the next step.

[0126] Step B: tert-butyl [3-(cyanomethyl)-3-[3-(1,3-dioxoisoindol-2-yl)-4-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]azetidine-1-sulfonyl]carbamate 2-{3-[3-(1,3-dioxoindole-2-yl)-4-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]azetidin-3-yl}acetonitrile (3 g, 5.4 mmol, 1.0 eq) was dissolved in dichloromethane (20 mL) at 80°C. After dissolving, triethylamine (1.64, 16.2 mmol, 2.0 eq) was added and stirred for 15 minutes. tert-Butyl (chlorosulfonyl)carbamate (1.75 g, 8.1 mmol, 1.5 eq) was added and stirred at room temperature for 2 hours. After the reaction was completed, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 * 3 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified on a silica gel column (dichloromethane: methanol = 10:1) to give tert-butyl [3-(cyanomethyl)-3-[3-(1,3-dioxoisoindol-2-yl)-4-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]azetidine-1-sulfonyl]carbamate (2.2 g, yield = 71%).

[0127] LC-MS (ESI), m / z: [M+H] + = 733.9.

[0128] Step C: tert-Butyl ((3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-1-yl)sulfonyl)(2,2,2-trifluoroethyl)carbamate To a solution of tert-butyl ((3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-1-yl)sulfonyl)carbamate (200 mg, 0.27 mmol, 1.0 eq) in N,N-dimethylformamide (5 mL) was added cesium carbonate (264 mg, 0.81 mmol, 3.0 eq) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (175 mg, 0.54 mmol, 2.0 eq), and the reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL*3). The combined organic phase was washed with saturated brine (20 mL). The combined organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the crude product. The crude product was purified through a Combiflush column (petroleum ether:ethyl acetate = 3:1) to give the product tert-butyl ((3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-1-yl)sulfonyl)(2,2,2-trifluoroethyl)carbamate (95 mg, yield = 43%) as a white solid.

[0129] LC-MS(ESI), m / z: [M+H] + =815.9.

[0130] Step D: 3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-N-(2,2,2-trifluoroethyl)azetidine-1-sulfonamide tert-Butyl ((3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-1-yl)sulfonyl)(2,2,2-trifluoroethyl)carbamate (95 mg, 0.12 mmol, 1.0 eq) was added to dichloromethane (5 ml) at room temperature, and boron trifluoride ethyl ether solution (0.5 ml) was added at 0 °C. The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was extracted with ethyl acetate and water, and the organic phase was dried and concentrated to give the crude product (50 mg, yield = 58%).

[0131] LC-MS(ESI), m / z: [M+H] + =615.9.

[0132] Step E: 3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)-N-(2,2,2-trifluoroethyl)azetidine-1-sulfonamide 3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-N-(2,2,2-trifluoroethyl)azetidine-1-sulfonamide (50 mg, 0.07 mmol, 1.0 eq) was dissolved in methanol (2 mL) at room temperature, and ethylenediamine (0.2 mL) was added in an ice bath to adjust the pH to 8–9. The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated and purified by Prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-HO (0.1% TFA), gradient 35%-40%, flow rate: 20 mL / min) to obtain the product (17.5 mg, yield = 55%).

[0133] LC-MS(ESI), m / z: [M+H] + =455.7.

[0134] 1 H-NMR(400 MHz, CD3OD) δ 8.58 (s, 1H), 8.40 (s, 1H), 7.34 (d, J = 3.6 Hz, 1H), 6.86 (d, J = 3.6 Hz, 1H), 4.39 (d, J = 9.3 Hz, 2H), 4.02 (d, J = 9.4 Hz, 2H), 3.68 (q, J = 9.1 Hz, 2H), 3.38 (s, 2H).

[0135] The following examples were prepared with reference to the experimental route and methods of Example 40: JPEG2026504901000069.jpg253160JPEG2026504901000070.jpg231160JPEG2026504901000071.jpg246160JPEG2026504901000072.jpg230160JPEG2026504901000073.jpg140161Example 55: 3-(cyanomethyl)-3-(3-hydroxy-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-N-(2,2,2-trifluoroethyl)azetidine-1-formamide JPEG2026504901000074.jpg83129Synthetic route: JPEG2026504901000075.jpg83162 Step A: 2-cyano-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)acetic acid ethyl ester 4-Chloro-7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidine (1 g, 3.5 mmol, 1.0 eq), ethyl 2-cyanoacetate (1196 mg, 10.6 mmol, 3.0 eq), and potassium carbonate (974 mg, 7 mmol, 2.0 eq) were added to N,N-dimethylformamide (10 mL) and stirred at 60 °C for 0.5 h. The solution was heated to 130 °C and stirred for 1 h. After the reaction was completed, it was diluted with water (50 mL), extracted with ethyl acetate (20 mL*3), the organic phase was dried and concentrated, evaporated to dryness, and purified with silica gel column (petroleum ether: ethyl acetate = 5:1) to obtain the product ethyl 2-cyano-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)acetate (1.1 g, yield = 88%).

[0136] LC-MS (ESI), m / z: [M+H] + = 361.0. Step B: Ethyl (2E)-3-amino-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)propan-2-enoate JPEG2026504901000077.jpg4787 Ethyl 2-cyano-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)acetate (1000 mg, 2.8 mmol, 1.0 eq) and triethylamine (310 mg, 3.0 mmol, 1.1 eq) were added to dry tetrahydrofuran (20 mL), and diisobutylaluminum hydride (1 M in THF, 8.0 mL) was added dropwise at 0°C. The solution was stirred at 25°C for 3 h. After the reaction was completed, it was diluted with water (50 mL), extracted with ethyl acetate (30 mL*3), the organic phase was dried and concentrated, evaporated to dryness, and purified on a silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain the product ethyl (2E)-3-amino-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)propan-2-enoate (700 mg, yield = 70.0%).

[0137] LC-MS (ESI), m / z: [M+H] + = 363.1.

[0138] Step C: 4-{7-[(2-methoxyethyl)trimethyl-{5}-silanyl]pyrrolo[2,3-d]pyrimidin-4-yl}-1H-pyrazole-3-alcohol JPEG2026504901000078.jpg4991 (2E)-3-amino-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)propan-2-enoate (700 mg, 1.9 mmol, 1.0 eq), water, and hydrazine (193 mg, 3.86 mmol, 2.0 eq) were added to dioxane (10 mL) and stirred at 80 °C for 2 h. The reaction mixture was concentrated, evaporated to dryness, and purified on a silica gel column (dichloromethane:methanol = 20:1) to give the product 4-{7-[(2-methoxyethyl)trimethyl-{5}-silanyl]pyrrolo[2,3-d]pyrimidin-4-yl}-1H-pyrazole-3-alcohol (600 mg, yield = 76%).

[0139] LC-MS (ESI), m / z: [M+H] + = 332.1.

[0140] Step D: 3-cyanomethylene-N-(2,2,2-trifluoroethyl)azetidine-1-formamide JPEG2026504901000079.jpg50912-(Azetidin-3-ylidene)acetonitrile hydrochloride (200 mg, 1.53 mmol, 1.0 eq) was dissolved in dichloromethane (8 ml). N,N-diisopropylethylamine (593 mg, 4.59 mmol, 3.0 eq) was added dropwise to the reaction mixture at 0°C. The reaction mixture was stirred at 25°C for 15 minutes. (2,2,2-Trifluoroethyl)phenylcarbamate (337 mg, 1.53 mmol, 1.0 eq) was then added to the reaction mixture. The reaction mixture was stirred at 25°C for 18 hours. After the reaction was completed, the reaction mixture was evaporated under reduced pressure until dry. The residue obtained by concentrating the reaction mixture was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain the product, 3-cyamethylene-N-(2,2,2-trifluoroethyl)azetidine-1-formamide (200 mg, yield = 95%) as a white solid.

[0141] LC-MS (ESI), m / z: [M+H] + =262.0.

[0142] Step E: 3-cyanomethyl-3-hydroxy-4-(7-(2-trimethylsilylethoxymethyl)-7H-pyrrolidin-4-ylpyrimidin-1-pyrazol-1-yl-N-(2,2,2-trifluoroethyl)azetidine-1-formamide 4-(2-Trimethylsilylethoxymethyl)pyrrolopyrrolidin-4-ylpyrazole (302 mg, 0.91 mmol, 1.0 eq) and 3-cyanomethylene-N-(2,2,2-trifluoroethyl)azetidine-1-formamide (200 mg, 0.91 mmol, 1.0 eq) were dissolved in N,N-dimethylformamide (8 mL). 1,8-Diazabicyclo[5.4.0]undecen-7-ene (92 mg, 0.91 mmol, 1.0 eq) was then added. The reaction mixture was incubated at 25°C for 18 hours. After the reaction was completed, water (20 mL) was added to the reaction mixture, which was then extracted with dichloromethane (20 mL * 3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate, and the reaction mixture was concentrated. The resulting residue was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to give the product, 3-cyanomethyl-3-hydroxy-4-(7-(2-trimethylsilylethoxymethyl)-7H-pyrrolidin-4-ylpyrimidin-1-pyrazol-1-yl-N-(2,2,2-trifluoroethyl)azetidine-1-formamide (100 mg, yield = 30%) as a yellow oil.

[0143] LC-MS (ESI), m / z: [M+H] + =551.0.

[0144] Step F: 3-cyanomethyl-3-(3-hydroxy-4-methylol-7H-pyrrolylpyrrolyl)-2,3-d-pyrrolidin-4-yl-1-pyrazolyl-N-(2,2,2-trifluoroethyl)azetidine-1-formamide JPEG2026504901000081.jpg881143-Cyanomethyl-3-hydroxy-4-(7-(2-trimethylsilylethoxymethyl)-7H-pyrrolidin-4-ylpyrimidin-1-pyrazol-1-yl-N-(2,2,2-trifluoroethyl)azetidine-1-formamide (150 mg, 0.27 mmol, 1.0 eq) was dissolved in dichloromethane (6 ml). In an ice bath, boron trifluoride-ethyl ether solution (2 mL) was added dropwise. The reaction mixture was stirred at 25°C for 2 h. After completion of the reaction, the mixture was evaporated to dryness under reduced pressure. The crude product, 3-cyanomethyl-3-(3-hydroxy-4-methylol-7H-pyrrolylpyrrolyl)-2,3-d-pyrrolidin-4-yl-1-pyrazolyl-N-(2,2,2-trifluoroethyl)azetidine-1-formamide (82 mg, crude) was obtained as a yellow oil.

[0145] LC-MS (ESI), m / z: [M+H] + =451.0.

[0146] Step D: 3-cyanomethyl-3-hydroxy-4-pyrrolidin-4-ylpyrimidin-2,3-dimethylpyrazol-1-yl-N-(2,2,2-trifluoroethyl)azetidine-1-formamide JPEG2026504901000082.jpg2531553-Cyanomethyl-3-(3-hydroxy-4-methylol-7H-pyrrolylpyrrolyl)-2,3-d-pyrrolidin-4-yl-1-pyrazolyl-N-(2,2,2-trifluoroethyl)azetidine-1-formamide (82 mg, 0.18 mmol, 10.0 eq) was dissolved in methanol (5 mL) and ethylenediamine (0.5 mL) was added in an ice bath. After completion of the reaction, water (20 mL) was added to the reaction solution, which was then extracted with dichloromethane (10 mL*3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The residue was purified by Prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-HO (0.1% FA), gradient 10%-40%, flow rate: 20 mL / min) to obtain the product (5.3 mg, yield = 7%).

[0147] LC-MS (ESI), m / z: [M+H] + =421.0.

[0148] 1 H-NMR(400MHz, CD3OD) δ 8.67 (s, 1H), 8.60 (s, 1H), 7.49 (d, J = 3.6 Hz, 1H), 7.05 (d, J = 3.6 Hz, 1H), 4.56 (d, J = 9.3 Hz, 2H), 4.28 (d, J = 9.3 Hz, 2H), 3.82 (d, J = 9.3 Hz, 2H), 3.48 (s, 2H).

[0149] The following examples were prepared with reference to the experimental route and methods of Example 55: JPEG2026504901000083.jpg253155JPEG2026504901000084.jpg63155Example 60: 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-formamide JPEG2026504901000085.jpg73104Synthetic route: JPEG2026504901000086.jpg88156 Step A: 4-Bromo-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylic acid methyl ester JPEG2026504901000087.jpg4388 To a solution of methyl 4-bromo-1H-pyrazole-3-carboxylate (5 g, 25 mmol, 1.0 eq) in acetonitrile (50 mL) was added potassium carbonate (6.78 g, 50 mmol, 2.0 eq) and p-methoxybenzyl chloride (7.69 g, 25 mmol, 1.0 eq). The reaction mixture was stirred at 25 °C for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure to give the product, 4-bromo-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylic acid methyl ester (5.88 g, crude) as a yellow solid.

[0150] LC-MS(ESI), m / z: [M+H] + =325.0.

[0151] Step B: 1-(4-Methoxybenzyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-3-carboxylic acid methyl ester To a solution of 4-bromo-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylic acid methyl ester (5.88 g, 18 mmol, 1.0 eq), bis(pinacolato)diboron (4.58 g, 18 mmol, 1.0 eq), and potassium acetate (5.292 g, 54 mmol l, 3.0 eq) in 1,4-dioxane (200 mL) was added [1,1'-bis(diphenylphosphinoyl)ferrocene]dichloropalladium(II) (1.316 g, 1.8 mmol, 0.1 eq). The reaction mixture was stirred at 90°C under nitrogen gas protection for 16 h, diluted with water (20 mL), extracted with ethyl acetate (20 mL*3), and the combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The crude product was purified through a Combiflash column (petroleum ether:ethyl acetate = 2:1) to give the crude product (3.74 g) as a yellow solid.

[0152] LC-MS(ESI), m / z: [M+H] + =373.1.

[0153] Step C: 1-(4-Methoxybenzyl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylic acid methyl ester To a solution of 1-(4-methoxybenzyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-3-carboxylic acid methyl ester (3.74 g, 10 mmol, 1.0 eq), 4-chloro-7-((2-trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (2.83 g, 10 mmol, 1.0 eq), and potassium carbonate (4.14 g, 30 mmol, 3.0 eq) in 1,4-dioxane was added bistriphenylphosphinepalladium dichloride (0.7 g, 1 mmol, 0.1 eq). The reaction mixture was stirred at 90°C under nitrogen gas protection for 16 h, diluted with water (20 mL), extracted with ethyl acetate (20 mL*3), and the combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The crude product was purified through a Combiflash column (dichloromethane:methanol = 20:1) to give the crude product (1.85 g) as a yellow solid.

[0154] LC-MS(ESI), m / z: [M+H]+ =494.1.

[0155] Step D: Methyl 4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylate JPEG2026504901000090.jpg51941-(4-Methoxybenzyl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylic acid methyl ester (1.85 g, 3.7 mmol, 1.0 eq) in methanol (20 mL) was added cerium ammonium nitrate (9.8 g, 18.5 mmol, 5.0 eq). The reaction mixture was stirred overnight at 25 °C. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL*3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The crude product was obtained and purified through a Combiflash column (dichloromethane:methanol=15:1) to give the crude product (900 mg, yield=65%) as a yellow solid.

[0156] LC-MS(ESI), m / z: [M+H] + =374.1.

[0157] Step E: 1-(1-(tert-butoxycarbonyl)-3-(cyanomethyl)azetidin-3-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylic acid methyl ester JPEG2026504901000091.jpg761354-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylic acid methyl ester (900 mg, 2.41 mmol, 1.0 eq), 1,8-diazobispyro[5.4.0]undec-7-ene (367 mg, 2.41 mmol, 1.0 eq) in N,N-dimethylformamide (5 mL) was added tert-butyl 3-(cyanomethylene)azetidine-1-carboxylate (472 mg, 2.41 mmol, 1.0 eq), and the reaction mixture was stirred at 25 °C overnight. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL*3). The combined organic phase was washed with saturated brine (20 mL). The combined organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the crude product. The crude product was purified through a Combiflash column (dichloromethane:methanol = 20:1) to give the product 1-(1-(tert-butoxycarbonyl)-3-(cyanomethyl)azetidin-3-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylic acid methyl ester (800 mg, yield = 58%) as a white solid.

[0158] LC-MS(ESI), m / z: [M+H] + =568.2.

[0159] Step F: tert-Butyl 3-(3-carbamoyl-4-(7-(2-(trimethylsilyl)ethoxy)-7h-pyrrolo[2,3-3-]pyrimidin-4-yl)-1h-pyrazol-1-yl)-3-(cyanomethyl)azepine-1-carboxylate JPEG2026504901000092.jpg75127 A solution of methyl 1-(1-(tert-butoxycarbonyl)-3-(cyanomethyl)azetidin-3-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylate (250 mg, 0.44 mmol, 1.0 eq) in ammonia and methanol (5 mL) was stirred at 25°C overnight. Evaporation to dryness under reduced pressure gave crude tert-butyl 3-(3-carbamoyl-4-(7-(2-(trimethylsilyl)ethoxy)-7H-pyrrolo[2,3-3-]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azepine-1-carboxylate (195 mg, yield=80%).

[0160] LC-MS(ESI), m / z: [M+H] + =553.2.

[0161] Step G: 1-(3-(cyanomethyl)azapyridin-3-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide JPEG2026504901000093.jpg67126 A solution of tert-butyl 3-(3-carbamoyl-4-(7-(2-(trimethylsilyl)ethoxy)-7H-pyrrolo[2,3-3-]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azepine-1-carboxylate (195 mg, 0.35 mmol, 1.0 eq) in 4 M hydrochloric acid-1,4-dioxane (5 mL) was stirred at 25°C for 2 h. Evaporation to dryness under reduced pressure gave crude 1-(3-(cyanomethyl)azapyridin-3-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (135 mg, crude).

[0162] LC-MS(ESI), m / z: [M+H] +=453.1.

[0163] Step H: 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azepin-3-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide JPEG2026504901000094.jpg811361-(3-(cyanomethyl)azapyridin-3-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (135 mg, 0.3 mmol, 1.0 eq) and triethylamine (60 mg, 0.6 mmol, 2.0 eq) in acetonitrile (5 mL) was added 2,2,2-trifluoroethane-1-sulfonyl chloride (54.3 mg, 0.3 mmol, 1.0 eq). The reaction was stirred at 25 °C for 2 h. The reaction was stirred at 25 °C overnight. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL*3). The combined organic phase was washed with saturated brine (20 mL). The combined organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the crude product. The crude product was purified through a Combiflash column (dichloromethane:methanol = 20:1) to give the product 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azepin-3-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (100 mg, yield = 56%) as a yellow solid.

[0164] LC-MS(ESI), m / z: [M+H] + =599.0.

[0165] Step I: 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azapyridin-3-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide JPEG2026504901000095.jpg901421-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azepin-3-yl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (100 mg, 0.17 mmol, 1.0 eq) in dichloromethane (5 mL) was added boron trifluoride-ethyl ether solution (0.5 mL). The reaction mixture was stirred at 25°C for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the product 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azapyridin-3-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (63 mg, crude) as a yellow solid.

[0166] LC-MS(ESI), m / z: [M+H] + =499.0.

[0167] Step J: 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azapyridin-3-yl)-4-(7h-pyrrolo[2,3-d]pyrimidin-4-yl)-1h-pyrazole-3-carboxamide JPEG2026504901000096.jpg831231-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azapyridin-3-yl)-4-(7-(methylol)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (63 mg, 0.13 mmol, 1.0 eq) in methanol (2 mL) was added ethylenediamine (0.2 mL). The reaction mixture was stirred at 25 °C for 2 h. The solution was concentrated in vacuo, and the reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The crude product was obtained, and the combined organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The residue was passed through Prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-HO (0.1% TFA), gradient 25%-30%, flow rate: 20 mL / min) to give the product (3.2 mg, yield = 5.2%).

[0168] LC-MS(ESI), m / z: [M+H] + =469.0.

[0169] 1 H-NMR(400MHz, D2O) δ 8.72 (s, 1H), 8.69 (s, 1H), 8.30 (s, 1H), 7.57 (d, J = 3.6 Hz, 1H), 6.74 (d, J = 3.5 Hz, 1H), 4.77 (d, J = 9.4 Hz, 2H), 4.46 (d, J = 9.3 Hz, 2H), 4.39 (q, J = 9.2 Hz, 2H), 3.60 (s, 2H).

[0170] The following examples were prepared with reference to the experimental route and methods of Example 60: JPEG2026504901000097.jpg238155JPEG2026504901000098.jpg230156JPEG2026504901000099.jpg124154Example 69: 2-[3-(3-amino-4-{6-[(1-methylpyrazol-4-yl)amino]pyrimidin-4-yl}pyrazol-1-yl)-1-methylsulfonylpropidin-3-yl]acetonitrile JPEG2026504901000100.jpg77105Synthetic route: JPEG2026504901000101.jpg35163 Step A: tert-Butyl [3-amino-4-(2-chloropyrimidin-4-yl)pyrazol-1-yl]carboxylate JPEG2026504901000102.jpg5186 tert-Butyl [3-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboralkane-2-yl)pyrazol-1-yl]carboxylate (418 mg, 2.85 mmol, 1.0 eq) in 1,4-dioxane (10 mL) was added 2,4-dichloropyrimidine (644.44 mg, 4.28 mmol, 1.5 eq), sodium carbonate (611 mg, 5.82 mmol, 2.0 eq), and dichlorobis(triphenylphosphine)palladium(II) (141.81 mg, 0.194 mmol, 0.07 eq) was added, and the reaction mixture was stirred in an oil bath at 110°C under nitrogen gas protection for 3 hours. After completion of the reaction, the reaction mixture was evaporated to dryness and purified on a silica gel column (petroleum ether:ethyl acetate=1:1) to obtain tert-butyl [3-amino-4-(2-chloropyrimidin-4-yl)pyrazol-1-yl]carboxylate (150 mg, yield=35%).

[0171] LC-MS (ESI), m / z: [M+H] + = 296.1.

[0172] Step B: 6-(3-amino-1H-pyrazol-4-yl)-N-1-methylpyrazol-4yl)pyrimidin-4-amine tert-Butyl [3-amino-4-(2-chloropyrimidin-4-yl)pyrazol-1-yl]carboxylate (100 mg, 0.33 mmol, 1.0 eq) and 1-methylpyrazol-4-amine (46.7 mg, 0.495 mmol, 1.5 eq) were dissolved in n-butanol (5 mL) at room temperature, followed by the addition of p-toluenesulfonic acid (170 mg, 0.99 mmol, 3.0 eq). The mixture was stirred at 110 °C for 4 h, then concentrated and evaporated to dryness. The product was purified on a silica gel column (dichloromethane:methanol = 10:1) to give the product 6-(3-amino-1H-pyrazol-4-yl)-N-1-methylpyrazol-4-yl)pyrimidin-4-amine (70 mg, yield = 80%).

[0173] LC-MS (ESI), m / z: [M+H] + = 257.0.

[0174] Step C: 2-[3-(3-amino-4-{6-[(1-methylpyrazol-4-yl)amino]pyrimidin-4-yl}pyrazol-1-yl)-1-methylsulfonylpropidin-3-yl]acetonitrile JPEG2026504901000104.jpg911386-(3-Amino-1H-pyrazol-4-yl)-N-1-methylpyrazol-4-yl)pyrimidin-4-amine (70 mg, 0.27 mmol, 1.0 eq) and 2-(1-methylsulfonylated aziridin-3-ylidene)acetonitrile (70.5 mg, 0.41 mmol, 1.5 eq) were dissolved in acetonitrile (5 mL). 1,8-Diazabicyclo[5.4.0]undec-7-ene (41.04 mg, 0.27 mmol, 1.0 eq) was added and the mixture was stirred at 60 °C for 3 h. The reaction mixture was concentrated, and the residue was purified by prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-HO (0.1% TFA), gradient 25%-30%, flow rate: 20 mL / min) to give the product, 2-[3-(3-amino-4-{6-[(1-methylpyrazol-4-yl)amino]pyrimidin-4-yl}pyrazol-1-yl)-1-methylsulfonylpropidin-3-yl]acetonitrile (2 mg, yield=2%).

[0175] LC-MS (ESI), m / z: [M+H] + = 429.0.

[0176] 1 H-NMR(400MHz, CD3OD) δ 8.66 (s, 1H), 8.39 (s, 1H), 8.05 (s, 1H), 7.66 (s, 1H), 7.00 (s, 1H), 4.47 (d, J = 9.2 Hz, 2H), 4.20 (d, J = 9.2 Hz, 2H), 3.91 (s, 3H), 3.51 (s, 2H), 3.04 (s, 3H).

[0177] The following examples were prepared with reference to the experimental route and methods of Example 69: JPEG2026504901000105.jpg253158JPEG2026504901000106.jpg253155JPEG2026504901000107.jpg244156JPEG2026504901000108.jpg49157Example 80: 2-(3-(3-amino-4-(6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-b]pyridazin-8-yl)-1H-pyridin-1-yl)-1-(ethylsulfonyl)azetidin-3-yl)acetonitrile JPEG2026504901000109.jpg83112Synthetic route: JPEG2026504901000110.jpg39164 Step A: 4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-3-amine tert-Butyl 3-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate (300 mg, 0.97 mmol, 1.0 eq), 8-bromo-6-chloroimidazo[1,2-b]pyridazine (338 mg, 1.46 mmol, 1.5 eq), sodium carbonate (305.8 mg, 2.9 mmol, 3.0 eq), and [1,1'-bis(diphenylphosphinoyl)ferrocene]dichloropalladium(II) (71 mg, 0.09 mmol, 0.1 eq) were dissolved in 1,4-dioxane (10 ml). The reaction mixture was stirred at 80 °C for 2 h. After completion of the reaction, the reaction mixture was evaporated under reduced pressure to dryness. The residue obtained by concentrating the reaction mixture was purified by column chromatography (DCM:MeOH=20:1) to obtain the product 4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-3-amine (131 mg, yield=58%) as a yellow oil.

[0178] LC-MS (ESI), m / z: [M+H] + =235.0.

[0179] Step B: 2-(3-(3-amino-4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-1-yl)-1-(ethylsulfonyl)azetidin-3-yl)acetylnitrile JPEG2026504901000112.jpg751174-(6-Chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-3-amine (131 mg, 0.59 mmol, 1.0 eq) and 2-(1-(ethylsulfonyl)azetidin-3-ylidene)acetylnitrile (104 mg, 0.59 mmol, 1.0 eq) were dissolved in acetonitrile (5 ml). 1,8-Diazepinebicyclo[5.4.0]undec-7-ene (85 mg, 0.59 mmol, 1.0 eq) was added dropwise to the reaction mixture and stirred at room temperature for 4 h. After the reaction was completed, ice water (20 ml) was added to the reaction mixture and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The residue was passed through Prep-HPLC (column model: gemini-C18 150 x 19 mm, 5 μm, mobile phase: ACN-HO (0.1% TFA), gradient 10%-55%, flow rate: 20 mL / min) to obtain the product (200 mg, yield = 80.1%).

[0180] LC-MS (ESI), m / z: [M+H] + =420.8.

[0181] 1 H-NMR(400MHz,CD3OD) δ 8.68 (s, 1H), 8.05 (s, 1H), 7.74 (s, 1H), 7.43 (s, 1H), 4.54 (d, J = 8.8 Hz, 2H), 4.16 (d, J = 9.0 Hz, 2H), 3.48 (s, 2H), 3.17 - 3.11 (m, 2H), 1.33 (t, J = 7.3 Hz, 3H).

[0182] Step C: 2-(3-(3-amino-4-(6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-b]pyridazin-8-yl)-1H-pyridin-1-yl)-1-(ethylsulfonyl)azepan-3-yl)acetonitrile JPEG2026504901000113.jpg901302-(3-(3-amino-4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-1-yl)-1-(ethylsulfonyl)azepan-3-yl)acetonitrile (88 mg, 0.21 mmol, 1.0 eq), (1-methyl-1H-pyrazol-4-yl)boronic acid (27 mg, 0.21 mmol, 1.0 eq), and sodium tert-butyl alcohol (61 mg, 0.63 mmol, 3.0 eq) in tolyl / methanol (1:1, 5 To a solution of (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenylyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (5 mg, 0.005 mmol, 0.03 eq) was added. The reaction mixture was stirred at 110 °C under nitrogen gas protection for 16 h. The mixture was then diluted with water (20 mL) and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The crude product was purified through a Combiflash column (petroleum ether:ethyl acetate = 5:1) to give the product (12.8 mg, yield = 13%).

[0183] LC-MS(ESI), m / z: [M+H] + =467.0.

[0184] 1H-NMR (400MHz, CD3OD) δ 8.67 (s, 1H), 8.31 (s, 1H), 8.14 (s, 1H), 8.07 (d, J = 1.2 Hz, 1H), 7.72 (d, J = 1.2 Hz, 1H), 7.67 (s, 1H), 4.61 (d, J = 9.2 Hz, 2H), 4.22 (d, J = 9.3 Hz, 2H), 4.01 (s, 3H), 3.54 (s, 2H), 3.18 (dt, J = 12.3, 6.2 Hz, 2H), 1.38 (m, 3H).

[0185] The following examples were prepared with reference to the experimental route and methods of Example 80: JPEG2026504901000114.jpg253158JPEG2026504901000115.jpg139157Example 86: 2-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2-aminopropyl)sulfonyl)azetidin-3-yl)acetonitrile JPEG2026504901000116.jpg79133Synthetic route: JPEG2026504901000117.jpg57158 Step A: Benzyl (1-hydroxylpropan-2-yl)carbamate JPEG2026504901000118.jpg2111010.0 g (133 mmol, 1.0 eq) of 2-aminopropane-1-alcohol was dissolved in 250 mL of tetrahydrofuran. 18.7 mL (133 mmol, 1.0 eq) of benzyl chloroformate and 22.2 mL (160 mmol, 1.2 eq) of triethylamine were slowly added dropwise in an ice bath and stirred overnight at room temperature. After confirming complete reaction by TLC, 200 mL of water was added and the mixture was extracted with ethyl acetate (100 mL*3). The organic layer was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The residue was purified by column chromatography (PE / EA = 10 / 1 to 1:1) to obtain the product (13.6 g, yield = 43%).

[0186] LC-MS (ESI), m / z: [M+H] + =210.0.

[0187] 1 H-NMR(400MHz, DMSO-d6) δ 7.40-7.03 (m, 5H), 7.02 (d, J = 8.0 Hz, 1H), 5.01 (s, 2H), 4.64 (t, J = 5.8 Hz, 1H), 3.48-3.55 (m, 1H), 3.32-3.37 (m, 1H), 3.17-3.23 (m, 1H), 1.02 (d, J = 6.8 Hz, 3H). Step B: 2-((benzyloxy)carbonyl)amino)propyl methanesulfonate 13.6 g (64.12 mmol, 1.0 eq) of benzyl (1-hydroxylpropan-2-yl)carbamate was dissolved in 50 mL of tetrahydrofuran. 8.8 g of methanesulfonyl chloride (76.94 mmol, 1.2 eq) and 20.7 mL of triethylamine (160.3 mmol, 2.5 eq) were slowly added dropwise in an ice bath and stirred overnight at room temperature. After confirming complete reaction by TLC, 100 mL of water was added and the mixture was extracted with ethyl acetate (100 mL x 3). The organic layer was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The residue was purified by column chromatography (PE / EA = 0 to 2:1) to obtain the product (13.4 g, yield = 60.1%).

[0188] LC-MS (ESI), m / z: [M+H] + =310.0.

[0189] 1H-NMR(400MHz, CDCl3) δ 7.41-7.32 (m, 5H), 5.13 (s, 2H), 4.89 (s, 1H), 4.29-4.27 (m, 1H), 4.21-4.15 (m, 1H), 4.09-4.06 (m, 1H), 3.00 (s, 3H), 1.29 (d, J = 7.0 Hz, 3H).

[0190] Step C: S-2-((benzyloxy)carbonyl)amino)propyl)ethyl sulfate 10.0 g (34.8 mmol, 1.0 eq) of 2-(benzyloxycarbonyl)aminomethylsulfonate propyl ester was dissolved in 60 mL of N,N-dimethylformamide, followed by the addition of 22.7 g (69.6 mmol, 2.0 eq) of cesium carbonate and 5.30 g (69.6 mmol, 2.0 eq) of sulfonioacetic acid. The mixture was stirred overnight at room temperature. After confirming the formation of the target product by LC-MS, 150 mL of water was added and the mixture was extracted three times with ethyl acetate. The organic layer was washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The residue was purified by column chromatography (PE / EA = 0 to 5:1) to obtain the product (3.34 g, yield = 28%).

[0191] LC-MS (ESI), m / z: [M+H] + =268.0.

[0192] 1 H-NMR(400MHz, CDCl3) δ 8.04 (s, 1H), 7.40-7.32 (m, 5H), 5.11 (s, 2H), 3.99-3.92 (m, 1H), 3.08-3.06 (m, 2H), 2.36 (s, 3H), 1.22 (d, J = 6.8 Hz, 3H).

[0193] Step D: Benzyl (1-(chlorosulfonyl)propan-2-yl)carbamate JPEG2026504901000121.jpg2210 A solution of N-chlorosuccinimide (6.68 g, 50 mmol, 3.0 eq) in acetonitrile (20 mL) was added to a solution of hydrochloric acid (2 M, 3.34 mL, 6.7 mmol, 0.5 eq) at 30 °C. The mixture was stirred for 15 min. Next, S-2-((benzyloxy)carbonyl)amino)propyl)ethyl sulfate (3.34 g, 13 mmol, 1.0 eq) was dissolved in acetonitrile (20 mL) and the solution was added dropwise to the mixture. The mixture was stirred at room temperature for 1 h. After TLC confirmed complete reaction, the reaction mixture was diluted with sodium bicarbonate (20 mL) and extracted with ethyl acetate (20 mL*3). The combined organic phase was washed with saturated brine (20 mL). The combined organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to obtain the crude product. The crude product was purified by Combiflash column (petroleum ether:ethyl acetate=5:1) to give the product benzyl (1-(chlorosulfonyl)propan-2-yl)carbamate (1.2 g, yield=29%) as a white solid.

[0194] LC-MS (ESI), m / z: [M+H] + =291.9.

[0195] Step E: Benzyl (1-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidin-1-yl)sulfonyl)propan-2-yl)carbamate JPEG2026504901000122.jpg761492-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (600 mg, 2.04 mmol, 1.0 eq) in N,N-dimethylformamide (10 mL) was added N,N-diisopropylethylamine (787 mg, 6.1 mmol, 3.0 eq) and benzyl (1-(chlorosulfonyl)propan-2-yl)carbamate (1.2 g, 2.04 mmol, 1.0 eq), and the reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by LC-MS, the reaction solution was rotary evaporated to dryness with an oil pump, and the crude product was purified by reverse-phase Combiflash column (acetonitrile:water=3:7) to give the product benzyl (1-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidin-1-yl)sulfonyl)propan-2-yl)carbamate (500 mg, yield=45%) as a pale yellow solid.

[0196] LC-MS (ESI), m / z: [M+H] + =549.9.

[0197] Step F: 2-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-(2-aminopropyl)sulfonyl)azetidin-3-yl)acetonitrile JPEG2026504901000123.jpg6713520 mg (0.036 mmol, 1.0 eq) benzyl (1-(3-(3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidin-1-yl)sulfonyl)propan-2-yl)carbamate was dissolved in 5 mL of methanol, followed by the addition of 19 mg (0.18 mmol, 5.0 eq) of 10% palladium on carbon, followed by stirring at room temperature for 3 h under a hydrogen gas atmosphere. After confirming the formation of the target product by LC-MS, the reaction mixture was rotary evaporated to dryness, purified (mobile phase A: 0.1% formic acid + 0.05% triethylamine; mobile phase B: acetonitrile; gradient: 30%-70% B, 55 min; flow rate: 70 mL / min), and then desalted and purified (95% acetonitrile:5% water, 5 min, then 90% acetonitrile:70% water:methanol, flow rate: 70 mL / min) to give a white solid (3.7 mg, yield = 24%).

[0198] LC-MS: (M+H) + ; m / z=416.0.

[0199] 1H-NMR (400MHz, CD3OD) δ 8.70 (s, 1H), 8.52 (s, 1H), 7.46 (d, J = 3.6 Hz, 1H), 6.98 (d, J = 4.0 Hz, 1H), 4.68-4.64 (m, 2H), 4.28 (d, J = 9.2 Hz, 2H), 3.65-3.58 (m, 1H), 3.55 (s, 2H), 3.40-3.34 (m, 1H), 3.29-3.23 (m, 1H), 1.31 (d, J =6.8 Hz, 3H).

[0200] The following examples were prepared with reference to the experimental route and methods of Example 86: JPEG2026504901000124.jpg157158Bioactivity experiment 1. Enzyme activity (IC50) test of compounds (1) ROCK1 / 2 test experiment method (a) On an Echo 655, 50 nL of diluted compound working solution was transferred per well to a reaction plate (784075, Greiner).

[0201] (b) The reaction plate was sealed with sealing film and centrifuged at 1000 g for 1 minute.

[0202] (c) A kinase solution was prepared.

[0203] (d) 5 μL of kinase solution was added to each well of the reaction plate, which was then sealed with a sealing film, centrifuged at 1000 g for 30 seconds, and left at room temperature for 10 minutes.

[0204] (e) STK2-substrate-biotin A mixture of kinase substrate and ATP was prepared.

[0205] (f) 5 μL of a mixture of STK2-substrate-biotin and ATP was added to the reaction plate, and the plate was centrifuged at 1000 g for 30 seconds to initiate the reaction.

[0206] (g) ROCK1 kinase was reacted at room temperature for 20 minutes, and ROCK2 kinase was reacted at room temperature for 30 minutes.

[0207] (h) A mixture of Sa-XL 665 (125 nM) and STK-antibody-Cryptate was prepared using HTRF test buffer.

[0208] (i) 10 μL of a mixture of Sa-XL 665 and STK-antibody-Cryptate was added to each well, centrifuged at 1000 g for 30 seconds, and incubated at room temperature for 1 hour.

[0209] The signals at 615 nm (Cryptate) and 665 nm (XL665) were read on an Envision 2104, and the degree of kinase activity was characterized using the signal intensity.

[0210] Kinase activity data were expressed as the alignment of the kinase activity of the test compound with that of the blank group (containing only DMSO), and IC50 values ​​were obtained by curve fitting with Prism software (GraphPad 7.0).

[0211] The specific test results are shown in Table 1 below: Table 1. Enzyme activity of compounds (IC 50 )) JPEG2026504901000125.jpg252155JPEG2026504901000126.jpg255154JPEG2026504901000127.jpg219155

[0212] As can be seen from the data in the table, the enzymatic activity of the compounds in the examples of the present invention was superior to that of commercially available ROCK inhibitors, ripasudil, netarsudil, and vermosudil. Some compounds have potent activity against ROCK1 / 2 kinase and can be used to regulate the treatment of diseases mediated by ROCK kinase.

[0213] wherein Ripasudil has the following structural formula: JPEG2026504901000128.jpg135136

[0214] 2. Cellular activity of compounds (IC 50 ) Test (Myosin light chain phosphorylation test by cell in situ Western blotting (In-CellWestern) ROCK induces cytoskeletal changes by phosphorylating two amino acid sites, T18 and S19, of myosin light chain. The rat smooth muscle cell line A7r5 was used. A7r5 cells were incubated in DMEM containing 10% FBS. Phospho-MLC-T18 / S19-specific antibodies and secondary detection antibodies were used to detect myosin light chain phosphorylation levels by in situ Western blotting. Cells treated with positive compounds served as positive controls, while a group treated with the compound's solvent alone served as a negative control. Cell nuclei were stained with DRAQ5 and used as an internal standard. Absolute IC50 values ​​were determined by fitting a nonlinear regression curve with a variable slope using GraphPad Prism 7.0 software.

[0215] On day 1, A7r5 cells were resuspended in serum-free medium and seeded at a density of 5,000 cells per well into a PDL-coated, 384-well, clear-bottom black plate. The cells were serum-starved for 4 hours, followed by incubation with the cells and compounds in serum-free medium for 1 hour. Each well was fixed with 50 μL of 8% PFA (paraformaldehyde) for 1 hour at room temperature. The liquid was discarded, and the cells were permeabilized with 90 μL of ice-cold methanol for 1 hour at 4°C. The plate was then washed three times with PBST (0.1% Tween 20-PBS) using an automated dispenser. After gently tapping the plate dry, 50 μL of blocking solution was added to each well and blocked for 1 hour at room temperature. Phospho-MLC-T18 / S19-specific antibodies were diluted 1:200 in blocking solution and added in 20 μL to each well. The wells were sealed with membranes and incubated overnight at 4°C.

[0216] The next day, the liquid in the wells was discarded, and the plate was washed five times with PBST using an automated dispenser. After gently tapping the plate dry, the secondary detection antibody was diluted 1:800 in blocking solution, and DRAQ5 was diluted 1:1000. 20 μL was added to each well and incubated at room temperature for 1 hour. The liquid in the wells was discarded, and the plate was washed three times with PBST using an automated dispenser, followed by three washes with ddH2O. After gently tapping the plate dry, it was scanned using a LICOR Odyssey near-infrared imaging scanner.

[0217] The specific test results are shown in Table 2 below: Table 2 Cellular activity of compounds (IC 50 ) JPEG2026504901000129.jpg94165

[0218] The compounds in the examples of the present invention have superior cellular activity to commercially available ROCK inhibitors, ripasudil, netarsudil and vermosudil, and also have stronger ability to induce cytoskeletal changes by phosphorylating two amino acid sites T18 / S19 of myosin light chain, thus the compounds of the present invention have broader application value.

[0219] 3. Effect of the compound on a glaucoma model induced by episcleral vein ablation in SD rats Test method: Male SD rats that passed quarantine inspection were collected and subjected to general anesthesia with isoflurane. The experimental eye (right eye) was topically anesthetized with proparacaine hydrochloride eye drops. The eyelids were opened, and the bulbar conjunctiva was incised along the superior corneoscleral rim. The fascia was bluntly peeled to expose the episcleral veins. Two episcleral veins adjacent to the superior rectus muscle and one episcleral vein at the temporal region were cauterized. Signs of successful cauterization were dilated blood vessels near the corneoscleral rim, loss of blood flow distal to the corneoscleral rim, and no bleeding. After surgery, the conjunctiva was sutured intermittently with 9-0 non-absorbable surgical sutures. Levofloxacin eye drops were administered after suturing. After the rats woke up, they were returned to their cages.

[0220] The wound was disinfected with iodine before and after surgery. After surgery, levofloxacin eye drops were instilled twice daily for 1 week. Carprofen was administered subcutaneously (10 mg / kg, 1 mL / kg) once 30 minutes before surgery and twice at 24-hour intervals after surgery.

[0221] Three days after model creation, animals were randomly assigned to groups based on intraocular pressure and body weight. Each group consisted of five rats, and the rats were randomly assigned to a normal control group, a model control group, a positive control group, and a test substance group. After model creation, the positive control (commercially available Rhopressa eye drops: 0.02% netarsudil eye drops) or the test substance (0.01% Example 2) was instilled into the conjunctival sac at a dose of one drop once daily for 14 consecutive days (a total of 14 doses). The first day of instillation was designated the first day of the study (D1).

[0222] During the experimental period, the general condition of the animals in each group was observed daily. After administration, body weight was measured at least once a week, and intraocular pressure was measured within 24 hours on days 1, 3, 7, 10, and 14 of administration. At the end of drug administration, animals were dissected as planned and subjected to ophthalmological examination. After euthanasia, gross anatomical observation and histopathological examination (BRN3A immunofluorescence staining: using a 20x objective lens at a distance of 2 mm from the optic nerve, the number of RGCs in one field of the retina was counted, and the number of RGCs in the next circle of the retina in one section) were performed.

[0223] Test results: Body weight: As shown in Figure 1, by the end of the experiment, the body weight of rats in each group showed a steady increasing trend, and there was no significant difference in the body weight and weight gain of rats among the groups (P>0.05).

[0224] Intraocular pressure: Data on intraocular pressure changes at different time points within 24 hours on days 1, 3, 7, 10, and 14 of administration showed that up to 8 hours after administration, the intraocular pressure of animals in each administration group tended to decrease, and at 8 hours it was significantly lower than that of the model control group. By 24 hours, the intraocular pressure tended to stabilize, and the intraocular pressure of animals in the model group was consistently higher than that of each administration group. The intraocular pressure data are shown in Figure 2.

[0225] Histopathology: As shown in Figure 3, compared with the model control group, the number of RGCs in the animals of the positive control group and test substance group was elevated to some extent.

[0226] As a result of the experiment, it was confirmed that the compound of the present invention has a certain ameliorative effect on glaucoma and a certain protective effect on rat RGCs when administered by instillation into the conjunctival sac of one eye of the SD rat glaucoma model induced by episcleral vein ablation once daily for 14 consecutive days.

[0227] Although the present invention has been described in connection with the above preferred embodiments, these embodiments are merely exemplary and for the purpose of illustration only, and on this basis various alternatives and improvements can be made to the present application, all of which fall within the scope of protection of the present application.

Claims

1. An azacycloalkane compound of formula I or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof: Here, R １ Ha-NHR １１ , -OR １１ , -SR １１ or -C(=O)NHR １１ where R １１ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～６ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, substituted or unsubstituted C ５～２０ Heteroaryl, substituted or unsubstituted C １～６ Alkyl acyl, substituted or unsubstituted C １～６ Alkylsulfonyl, substituted or unsubstituted C ５～２０ Heteroarylacyl, and substituted or unsubstituted C １～６ alkoxy; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , -C(=O)R ３３ , hydroxyl, sulfhydryl, substituted C ３～８ Cycloalkyl, substituted C ３～８ Heterocyclyl, substituted C ６～２０ Aryl and substituted C ５～２０ selected from the group consisting of heteroaryl; R ２ is hydrogen, halogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C １～６ Haloalkyl, substituted or unsubstituted C １～６ Alkoxy, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ３ is cyano, -CONH ２ and carboxy; L is -S(=O) ２ -, -C(=O)-, -CH ２ - and -S(=O)(=N)R Ｌ -, where R Ｌ represents hydrogen and substituted or unsubstituted C １～８ alkyl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ４ is halogen, substituted or unsubstituted C １～８ Alkyl, —NR ３１ R ３２ , substituted or unsubstituted C １～８ Alkoxy, substituted or unsubstituted C ２－８ Alkenyl, substituted or unsubstituted C ２－８ Alkynyl, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, hydrazide group, C １～８ Alkylsulfonyl, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, -COR ３３ , carboxy, and sulfhydryl; A １ is selected from C atoms; W １ , W ２ , W ３ , W ４ and W ５ are each independently selected from a C atom or a N atom, and W １ , W ２ , W ３ , W ４ and W ５ where one, two or three are N atoms; the dotted circle indicates a bond that forms an aromatic ring; R ２１ represents hydrogen, halogen, -NR ３１ R ３２ , substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ２２ represents hydrogen, halogen, -NR ３１ R ３２ , substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ２３ represents hydrogen, halogen, -NR ３１ R ３２ , substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ２４ represents hydrogen, halogen, -NR ３１ R ３２ , substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ２５ represents hydrogen, halogen, -NR ３１ R ３２ , substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; Or, R ２４ , R ２４ W is bonded ４ , and R ２５ , and R ２５ W is bonded ５ are tied together, and A １ , W １ , W ２ , W ３ , W ４ and W ５ forming a 5-membered aryl, cycloalkyl, heteroaryl or heterocycloalkylcyclyl fused to a 6-membered ring consisting of R ３１ and R ３２ are each independently hydrogen, sulfonamido, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, hydroxyl, —C(═O)OR ３４ and sulfhydryl; R ３３ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Alkoxy, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ３４ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Haloalkyl, substituted or unsubstituted C １～８ Alkoxy, and substituted or unsubstituted C ３～８ cycloalkyl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy, and sulfhydryl; n is 1, 2 or 3; wherein the compound is not:

2. R １ is -NHR １１ where R １１ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～６ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, substituted or unsubstituted C ５～２０ Heteroaryl, substituted or unsubstituted C １～６ Alkyl acyl, substituted or unsubstituted C １～６ Alkylsulfonyl, substituted or unsubstituted C ５～２０ Heteroarylacyl, and substituted or unsubstituted C １～６ alkoxy; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , -C(=O)R ３３ , hydroxyl, sulfhydryl, substituted C ３～８ Cycloalkyl, substituted C ３～８ Heterocyclyl, substituted C ６～２０ Aryl and substituted C ５～２０ selected from the group consisting of heteroaryl; Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, hydroxyl, —C(═O)OR ３４ and sulfhydryl; R ３３ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Alkoxy, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ３４ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Haloalkyl, substituted or unsubstituted C １～８ Alkoxy, and substituted or unsubstituted C ３～８ cycloalkyl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy, and sulfhydryl; 2. The azacycloalkane compound according to claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof.

3. R １１ represents hydrogen and substituted or unsubstituted C １～６ alkyl, and the substituents are selected from the group consisting of -NR ３１ R ３２ , -C(=O)R ３３ and substituted or unsubstituted C ３～８ 3. The azacycloalkane compound of claim 2, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, selected from the group consisting of heterocyclyls.

4. R １１ is a substituted or unsubstituted C １～６ alkyl, and the substituents are selected from the group consisting of -NR ３１ R ３２ where R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, and substituted or unsubstituted C ３～８ 3. The azacycloalkane compound of claim 2, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, selected from the group consisting of heterocyclyls.

5. R １１ is a substituted or unsubstituted C １～６ alkyl, and the substituent is selected from the group consisting of -C(=O)R ３３ where R ３３ are independently substituted or unsubstituted C １～６ Alkyl, and substituted or unsubstituted C ３～８ 3. The azacycloalkane compound of claim 2, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, selected from the group consisting of heterocyclyls.

6. R １ は、-NH ２ 、-NHC(=O)CH ３ 、 ****c1 3. The azacycloalkane compound of claim 2, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, selected from the group consisting of:

7. R １ is -OR １１ where R １１ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～６ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, substituted or unsubstituted C ５～２０ Heteroaryl, substituted or unsubstituted C １～６ Alkyl acyl, substituted or unsubstituted C １～６ Alkylsulfonyl, substituted or unsubstituted C ５～２０ Heteroarylacyl, and substituted or unsubstituted C １～６ alkoxy; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , -C(=O)R ３３ , hydroxyl, and sulfhydryl; Preferably, R １１ represents hydrogen and substituted or unsubstituted C １～６ alkyl, and the substituents are selected from the group consisting of -NR ３１ R ３２ , -C(=O)R ３３ and substituted or unsubstituted C ３～８ heterocyclyl; Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, hydroxyl, —C(═O)OR ３４ and sulfhydryl; R ３３ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Alkoxy, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ３４ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Haloalkyl, substituted or unsubstituted C １～８ Alkoxy, and substituted or unsubstituted C ３～８ cycloalkyl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ 2. The azacycloalkane compound of claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy, and sulfhydryl.

8. R １ is -C(=O)NHR １１ where R １１ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～６ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, substituted or unsubstituted C ５～２０ Heteroaryl, substituted or unsubstituted C １～６ Alkyl acyl, substituted or unsubstituted C １～６ Alkylsulfonyl, substituted or unsubstituted C ５～２０ Heteroarylacyl, and substituted or unsubstituted C １～６ alkoxy; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , -C(=O)R ３３ , hydroxyl, and sulfhydryl; Preferably, R １１ represents hydrogen and substituted or unsubstituted C １～６ alkyl, and the substituents are selected from the group consisting of -NR ３１ R ３２ , -C(=O)R ３３ and substituted or unsubstituted C ３～８ heterocyclyl; Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ２～６ Alkenyl, substituted or unsubstituted C ２～６ Alkynyl, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, hydroxyl, —C(═O)OR ３４ and sulfhydryl; R ３３ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Alkoxy, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ３４ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Haloalkyl, substituted or unsubstituted C １～８ Alkoxy, and substituted or unsubstituted C ３～８ cycloalkyl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy, and sulfhydryl; 2. The azacycloalkane compound according to claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof.

9. R ２ is selected from hydrogen, and R ３ is selected from cyano; or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof according to claim 1 .

10. ****c2 Here, in the structures A1 to A7, R ７ are each independently hydrogen, halogen, or —NR ３１ R ３２ and substituted or unsubstituted C ５～２０ heteroaryl; Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ６～２０ Aryl, substituted or unsubstituted C ５～２０ Heteroaryl, and substituted or unsubstituted C ３～８ heterocyclyl; Here, the substituents are halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy, and sulfhydryl; 2. The azacycloalkane compound according to claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof.

11. R ７ are each independently hydrogen, —NR ３１ R ３２ and substituted or unsubstituted C ５～２０ heteroaryl; Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, and substituted or unsubstituted C ５～２０ selected from the group consisting of heteroaryl; where the substituent is C １～８ 11. The azacycloalkane compound according to claim 10, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer or prodrug thereof, wherein the azacycloalkane compound is selected from the group consisting of alkyl, aryl, arylsulfonyl ...

12. ****c3 Here, in the A1 structure, R ７ 11. The azacycloalkane compound of claim 10, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, wherein is hydrogen.

13. ****c4 Here, in the A2 structure, R ７ Ha-NR ３１ R ３２ or substituted or unsubstituted C ５～２０ is heteroaryl, Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ５～２０ Heteroaryl, and substituted or unsubstituted C ３～８ heterocyclyl; where the substituent is C １～８ selected from the group consisting of alkyl and hydroxyl; ****c5 Here, in the A3 structure, R ７ Ha-NR ３１ R ３２ or substituted or unsubstituted C ５～２０ is heteroaryl, Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ５～２０ Heteroaryl, and substituted or unsubstituted C ３～８ heterocyclyl; where the substituent is C １～８ selected from the group consisting of alkyl and hydroxyl; ****c6 Here, in the A4 structure, R ７ Ha-NR ３１ R ３２ or substituted or unsubstituted C ５～２０ is heteroaryl, Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ５～２０ Heteroaryl, and substituted or unsubstituted C ３～８ heterocyclyl; where the substituent is C １～８ selected from the group consisting of alkyl and hydroxyl; 11. The azacycloalkane compound according to claim 10, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof.

14. ****c7 Here, in the A5 structure, R ７ Ha-NR ３１ R ３２ or substituted or unsubstituted C ５～２０ is heteroaryl, Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, or substituted and unsubstituted C ５～２０ selected from the group consisting of heteroaryl; where the substituent is C １～８ selected from the group consisting of alkyl and hydroxyl; ****c8 Here, in the A6 structure, R ７ is halogen, -NR ３１ R ３２ or substituted or unsubstituted C ５～２０ is heteroaryl, Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, and substituted or unsubstituted C ５～２０ selected from the group consisting of heteroaryl; where the substituent is C １～８ selected from the group consisting of alkyl and hydroxyl; ****c9 Here, in the A7 structure, R ７ Ha-NR ３１ R ３２ or substituted or unsubstituted C ５～２０ is heteroaryl, Here, R ３１ and R ３２ are each independently hydrogen, substituted or unsubstituted C １～６ Alkyl, and substituted or unsubstituted C ５～２０ selected from the group consisting of heteroaryl; where the substituent is C １～８ 11. The azacycloalkane compound of claim 10, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, wherein the azacycloalkane compound is selected from the group consisting of alkyl and hydroxyl.

15. ( ７ は!\H ３ 、!\H ２ ( ２ ௨、|、、 ****c10 The azacycloalkane compound according to any one of claims 10 to 11 or 13 to 14, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof.

16. L is -S(=O) ２ -, -C(=O)- and -S(=O)(=N)R Ｌ -, where R Ｌ is hydrogen and C １～６ 2. The azacycloalkane compound of claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, selected from the group consisting of alkyl.

17. R ４ is a substituted or unsubstituted C １～８ Alkyl, —NR ３１ R ３２ , substituted or unsubstituted C １～８ Alkoxy, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, hydrazide group, C １～８ Alkylsulfonyl, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, -COR ３３ , carboxy, and sulfhydryl; Here, R ３１ and R ３２ are each independently hydrogen, sulfonamido, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C １～６ Alkyl, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, cyano, hydroxyl, —C(═O)OR ３４ and sulfhydryl; R ３３ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Alkoxy, substituted or unsubstituted C ３～８ Cycloalkyl, substituted or unsubstituted C ３～８ Heterocyclyl, substituted or unsubstituted C ６～２０ Aryl, and substituted or unsubstituted C ５～２０ heteroaryl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ Heteroaryl, cyano, -NR ３１ R ３２ , hydroxyl, carboxy, and sulfhydryl; R ３４ is hydrogen, substituted or unsubstituted C １～８ Alkyl, substituted or unsubstituted C １～８ Haloalkyl, substituted or unsubstituted C １～８ Alkoxy, and substituted or unsubstituted C ３～８ cycloalkyl; the substituents are selected from the group consisting of halogen, C １～８ Alkyl, C １～８ Haloalkyl, C １～８ Alkoxy, C ３～８ Cycloalkyl, C ３～８ Heterocyclyl, C ６～２０ Aryl, C ５～２０ 2. The azacycloalkane compound of claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotopically labeled compound, isomer, or prodrug thereof, selected from the group consisting of heteroaryl, cyano, hydroxyl, carboxy, and sulfhydryl.

18. The azacycloalkane compound is selected from the group consisting of: ****24－28 2. The azacycloalkane compound according to claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof.

19. The azacycloalkane compound is selected from the group consisting of: ****29－30 2. The azacycloalkane compound according to claim 1, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof.

20. A pharmaceutical composition comprising the azacycloalkane compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer or prodrug thereof, and a pharmaceutically acceptable carrier.

21. Use of the azacycloalkane compound according to any one of claims 1 to 19 or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof, and the pharmaceutical composition according to claim 20, in the preparation of a medicament for treating a disease mediated by Rho kinase.

22. 22. The use according to claim 21, wherein the disease mediated by Rho kinase is asthma, cancer, glaucoma, insulin resistance, renal failure, neurodegeneration or osteoporosis.

23. A method for treating a disease mediated by Rho kinase, comprising administering to a patient in need thereof a therapeutically effective amount of the azacycloalkane compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt, solvate, active metabolite, crystalline polymorph, isotope-labeled compound, isomer, or prodrug thereof, or the pharmaceutical composition according to claim 20.

24. 24. The method of claim 23, wherein the Rho kinase-mediated disease is asthma, cancer, glaucoma, insulin resistance, renal failure, neurodegeneration, or osteoporosis.

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