Azacycloalkane compound, pharmaceutical composition and application of azacycloalkane compound

Abstract

The invention relates to an azacycloalkane compound, a pharmaceutical composition and application of the azacycloalkane compound. The azacycloalkane compound has a structural formula as shown in a formula (I). The azacycloalkane compound and the pharmaceutical composition thereof provided by the invention have remarkable Rho kinase inhibition activity, and the enzymatic activity and the cell activity of the azacycloalkane compound are superior to those of existing ROCK inhibitors such as Ripasubl, Netarsubl and belumosubl, so that the azacycloalkane compound and the pharmaceutical composition thereof have very high application potential. # imgabs0 #

Description

A kind of azacycloalkane compound, pharmaceutical composition and use thereof Technical Field

[0001] The present invention provides an azacycloalkane compound, a stereoisomer comprising the compound, a pharmaceutical composition, and uses of the compound and the pharmaceutical composition. The compound is an inhibitor targeting ROCK and can be used to regulate Rho kinase-mediated diseases. Background Art

[0002] The Rho kinase / ROCK (Rho-associated kinase) signaling pathway induces cytoskeletal reorganization, cell migration, adhesion, and stress fiber formation, and is associated with a variety of physiological functions. The ROCK family includes 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 acts on the E-cadherin complex by binding to the E-cadherin scaffolding protein p120-catenin. ROCK1 is localized to microtubule organizing centers and the edges of pseudopodia in motile cells, implicated in cell migration. ROCK2 is primarily localized in the cytoplasm and localized to the plasma membrane via its C-terminal region, where it associates with vimentin and actin tension fibers.

[0003] ROCK mediates many pathophysiological signals and is related to various physiological functions such as endothelial permeability, tissue contraction and growth. ROCK inhibitors have potential application value in diseases including asthma, cancer, glaucoma, insulin resistance, renal failure, neuronal degeneration and osteoporosis. Two ROCK1 / ROCK2 inhibitors, Ripasudil and Netarsudil eye drops, have been approved for the treatment of glaucoma, and a ROCK2 inhibitor, belumosudil, has been approved for the treatment of chronic graft-versus-host disease (cGVHD). However, the activity of existing marketed ROCK inhibitors is weak, and the function and clinical application value of ROCK are far from being fully developed. Therefore, it is urgent to develop a drug targeting ROCK to address unmet clinical needs.

[0004] Summary of the Invention

[0005] The present application develops a highly active azacycloalkane compound with a new skeleton targeting ROCK and a pharmaceutical composition thereof for regulating Rho kinase-mediated diseases.

[0006] The present application provides an azacycloalkane compound represented by Formula I or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof:

[0007] Another aspect of the present invention is to provide a method for preparing a compound as shown in Formula I, an isotope isomer or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof, and its use in treating ROCK-mediated diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 shows the changes in body weight of animals in each group during the experimental period;

[0009] Figure 2 shows the 24-hour intraocular pressure change curves of the animals in each group at different times after administration of D1;

[0010] FIG3 shows the RGCs results of animals in each group at the end of the experiment. DETAILED DESCRIPTION

[0011] The present invention will be further described in detail below through the following examples, through which the features and advantages of the present invention will become more clearly understood.

[0012] The word “exemplary” is used exclusively herein to mean “serving as an example, example, or illustration.” Any embodiment described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments.

[0013] In addition, the technical features involved in different embodiments of the present application described below can be combined with each other as long as they do not conflict with each other.

[0014] The following examples are provided for illustrative purposes and should not be construed as limiting the present invention in any manner. One skilled in the art will readily recognize that a variety of noncritical parameters can be changed or modified to yield essentially the same results.

[0015] definition

[0016] Unless otherwise defined, all technical and scientific terms herein have the same meanings as commonly understood by those skilled in the art to which the claimed subject matter belongs. Unless otherwise indicated, all patents, patent applications, and publications cited herein are incorporated by reference in their entirety. When trade names appear in this document, they are intended to refer to the corresponding commercial products or their active ingredients.

[0017] It should be understood that the above summary and the following detailed description are exemplary and explanatory only and do not limit the subject matter of the present invention in any way. In this application, it must be noted that unless otherwise clearly indicated, the singular forms used in this specification and claims include the plural forms of the referents. It should also be noted that unless otherwise indicated, the use of "or" and "or" means "and / or". In addition, the use of the term "including" and other forms, such as "comprising", "including" and "containing" are not limiting.

[0018] Definitions of standard chemical terms can be found in the literature, including Advanced Organic Chemistry by Carey and Sundberg. th Ed., Vol. A (2000) and B (2001), Plenum Press, New York. Unless otherwise indicated, conventional methods within the skill of the art, such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacological methods, are employed. Unless specifically defined, nomenclature and laboratory procedures and techniques in analytical chemistry, organic synthetic chemistry, and medicinal and pharmaceutical chemistry are known to those skilled in the art. Standard techniques can be used for chemical syntheses, chemical analyses, 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 filtration). For example, reactions and purification techniques can be performed using kits with manufacturer's instructions, or according to methods known in the art, or as described herein. In general, the aforementioned techniques and steps can be performed using conventional methods well known in the art and described in various general or more specific literature, which are cited and discussed herein.

[0019] When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents that would result if the formula were written from right to left. For example, CH2O is equivalent to OCH2.

[0020] The term "substituted or unsubstituted" includes both "substituted" and "unsubstituted", where "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, as long as the valence state of the specific atom is normal and the compound after substitution is stable; "unsubstituted" means that the hydrogen atoms on the specific atom are not replaced by a substituent. For example, "substituted or unsubstituted ethyl" (for example, when the substituent is a halogen) includes unsubstituted (-CH2CH3), monosubstituted (such as -CH2CH2F), polysubstituted (such as -CHFCH2F, -CH2CHF2, etc.) or fully substituted (-CF2CF3). It will be understood by those skilled in the art that for any group containing one or more substituents, no substitution or substitution pattern that is sterically impossible to exist and / or cannot be synthesized will be introduced. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms on the same atom are replaced.

[0021] When any variable (e.g., R) occurs more than once in a composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may optionally be substituted with up to two Rs, and each instance of R has independent options. In addition, combinations of substituents and / or their variants are permitted only if such combinations result in stable compounds. The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes both the occurrence of the event or circumstance and the non-occurrence of the event or circumstance.

[0022] The C m～n It means that there are m to n carbon atoms in the part. For example, the “C 1～8 " group means that the part has 1-8 carbon atoms, that is, the group contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms...8 carbon atoms. Therefore, for example, "C 1～8 "Alkyl" refers to an alkyl group containing 1 to 8 carbon atoms, that is, the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl...octyl, etc. The numerical ranges herein, such as "1-8", refer to each integer in the given range, for example, "1-8 carbon atoms" means that the group can 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.

[0023] The term "alkyl" refers to an optionally substituted straight chain or optionally substituted branched saturated aliphatic hydrocarbon group that is attached to the rest of the molecule by a single bond. "Alkyl" herein can have 1 to about 8 carbon atoms, for example, 1 to 6 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. Examples of "alkyl" herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, and the like, as well as longer alkyl groups such as heptyl and octyl, and the like. When a group defined herein, such as "alkyl," appears in a numerical range, for example, "C1-8 alkyl" refers to an alkyl group that can be composed of 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. For another example, "C1-4 alkyl" refers to an alkyl group that can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, or 4 carbon atoms. Alkyl groups herein also include those where no numerical range is specified.

[0024] The term "alkenyl" refers to an optionally substituted straight chain or optionally substituted branched monovalent hydrocarbon group having at least one C=C double bond. The alkenyl group has, but is not limited to, 2 to about 8 carbon atoms, such as 2 to about 6 carbon atoms, 2 to about 4 carbon atoms. The double bonds in these groups can be in cis or trans conformations and should be understood to include the two isomers. Examples of alkenyl groups include, but are not limited to, vinyl (CH=CH2), 1-propenyl (CH2CH=CH2), isopropenyl (C(CH3)=CH2), butenyl and 1,3-butadienyl, etc. When a numerical range appears in the alkenyl group defined herein, for example, "C2-8 alkenyl" refers to an alkenyl group that can be composed of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, or 8 carbon atoms, and the alkenyl group herein also encompasses the case where no numerical range is specified.

[0025] The term "alkynyl" refers to an optionally substituted straight or branched monovalent hydrocarbon group having at least one C≡C triple bond. The alkynyl group has, but is 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 groups herein include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and 1,3-butadiynyl. When a numerical range appears in the alkynyl group defined herein, for example, "C2-8 alkynyl" refers to an alkynyl group that can be composed of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, or 8 carbon atoms. The alkynyl group herein also encompasses situations where no numerical range is specified.

[0026] The term "cycloalkyl" refers to a non-aromatic carbon ring, including a saturated carbocyclic ring (such as a cycloalkyl group) or an unsaturated carbocyclic ring (such as a cycloalkenyl group). Carbocyclic rings include monocarbocyclic rings (having one ring), for example, a monocyclic cycloalkyl group; bicarbocyclic rings (having two rings), for example, a bicyclic cycloalkyl group; and polycarbocyclic rings (having more than two rings). The rings may be bridged or spirocyclic. Carbocyclic rings (such as cycloalkyl or cycloalkenyl groups) may have 3 to 8 carbon atoms, for example, 3 to about 6 ring-forming carbon atoms or 3 to about 5 ring-forming carbon atoms.

[0027] The term "aryl" refers to an optionally substituted aromatic hydrocarbon group having 6 to about 20, such as 6 to 12 or 6 to 10, ring-forming carbon atoms, which can be a monocyclic aromatic group, a bicyclic aromatic group, or a polycyclic aromatic group. A bicyclic aromatic group or a polycyclic aromatic group can be a monocyclic aromatic group fused to another independent ring, such as an alicyclic ring, a heterocyclic ring, an aromatic ring, or an aromatic heterocyclic ring. Non-limiting examples of monocyclic aromatic groups include monocyclic aromatic groups having 6 to about 12, 6 to about 10, or 6 to about 8 ring-forming carbon atoms, such as phenyl; bicyclic aromatic groups are exemplified by naphthyl; and polycyclic aromatic groups are exemplified by phenanthrenyl, anthracenyl, and azulenyl.

[0028] The term "heteroaryl" refers to an arbitrarily substituted heteroaryl group comprising from about 5 to about 20, such as 5 to 12 or 5 to 10 skeleton ring atoms, wherein at least one (such as 1-4, 1-3, 1-2) ring atoms is a heteroatom, and the heteroatoms are independently selected from the heteroatoms of oxygen, nitrogen, sulfur, phosphorus, silicon, selenium and tin, but are not limited thereto. Heteroaryl groups include monocyclic heteroaryl groups (having one ring), bicyclic heteroaryl groups (having two rings) or polycyclic heteroaryl groups (having more than two rings). In embodiments where two or more heteroatoms appear 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. Bicyclic heteroaryl groups or polycyclic heteroaryl groups may be a monocyclic heteroaryl group fused to other independent rings, such as alicyclic, heterocyclic, aromatic, or aromatic heterocyclic rings (collectively referred to as fused ring heteroaryl groups). Non-limiting examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, quinolyl, isoquinolyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, and the like.

[0029] The term "heterocyclyl" refers to a non-aromatic heterocycle, including a saturated heterocycle or an unsaturated heterocycle (containing an unsaturated bond), which does not have a completely conjugated π-electron system and can be classified as a monocyclic, fused polycyclic, bridged or spirocyclic system without aromaticity. One or more (e.g., 1-4, 1-3, 1-2) of the atoms forming the ring are heteroatoms, such as oxygen, nitrogen or sulfur atoms. The heterocycle can include a monocyclic heterocycle (having one ring), a bicyclic heterocycle (having two bridged rings) or a polycyclic heterocycle (having more than two bridged rings); spirocycles are also included. The heterocyclic group can have 3 to about 20, such as 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 groups include oxiranyl, thioranyl, aziridinyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, pyrrolidinyl, oxazolidinyl, tetrahydropyrazolyl, pyrrolinyl, dihydrofuranyl, dihydrothiophenyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyranyl, tetrahydropyranyl, dihydrothiopyranyl, azepanyl, oxepanyl, thiepanyl, oxazabicyclo[2.2.1]heptyl, and azaspiro[3.3]heptyl, and the like.

[0030] The term "halo" or "halogen" refers to an optionally substituted group (such as an alkyl, alkenyl, alkynyl, alkoxy group, etc.) in which at least one hydrogen atom is replaced with a halogen (such as fluorine, chlorine, bromine, iodine, or a combination thereof). In some embodiments, two or more hydrogen atoms are replaced with halogen atoms that are identical to each other (e.g., difluoromethyl, trifluoromethyl); in other embodiments, two or more hydrogen atoms are replaced with halogen atoms that are not identical to each other (e.g., 1-chloro-1-fluoro-1-iodoethyl).

[0031] The term "alkoxy" refers to an alkyl ether group (O-alkyl). Non-limiting examples of alkoxy include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, and the like.

[0032] The term "alkyl acyl" refers to a group consisting of an alkyl group connected to a -CO- group. Non-limiting examples of this term include formyl, acetyl, propionyl, butyryl, etc. For example, the term "C 1-6 "Alkyl acyl" refers to C 1-6 An alkyl group connected to -CO-. For example, the term "C 1-4 "Alkyl acyl" refers to C 1-4 A group formed by connecting an alkyl group to a -CO- group.

[0033] The term "alkylsulfonyl" refers to a group consisting of an alkyl group connected to -SO2-, and non-limiting examples of the term include methylsulfonyl, ethylsulfonyl, propanesulfonyl, butanesulfonyl, etc. For example, the term "C 1-6 "Alkylsulfonyl" refers to C 1-6 An alkyl group connected to -SO2-. For example, the term "C 1-4 "Alkylsulfonyl" refers to C 1-4 A group formed by connecting an alkyl group to a -SO2- group.

[0034] The term "heteroaryl acyl" refers to a group consisting of a heteroaryl group linked to a -CO- group. For example, the term "C 5～20 "Heteroarylsulfonyl" refers to C 5～20 A group consisting of a heteroaryl group and a -CO- group. 5～20 "Heteroaryl" is as defined above.

[0035] Other group terms herein include: "hydroxy" refers to an -OH group, "thiol" refers to a -SH group, "cyano" refers to a -CN group, and "carboxyl" refers to a -COOH group.

[0036] The term "membered" refers to the number of atoms that make up the ring. For example, pyridine is a six-membered ring, and pyrrole is a five-membered ring.

[0037] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0038] The term "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical component or agent, namely a "carrier", which facilitates the introduction of the compound into cells or tissues, including but not limited to stabilizers, diluents, suspending agents, thickening agents and / or excipients.

[0039] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological efficacy of the free acid and free base of the specified compound and has no adverse biological or other effects. Unless otherwise indicated, salts herein may include 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, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, and dicyclohexylamine. 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, and phosphoric acid. 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, benzenesulfonic acid, p-toluenesulfonic acid, etc. Non-limiting examples of salts formed with basic 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.

[0040] 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 base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or a mixture of the two. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

[0041] The term "solvate" refers to a physical aggregate formed by a compound of the present invention and one or more solvent molecules, which physical aggregate includes varying degrees of ionic and covalent bonds, such as hydrogen bonds. It has been shown that such solvates can be isolated, for example, when one or more solvent molecules are mixed in the crystal lattice. A "solvate" includes two parts: a solvent phase and a separable solvate. There are many examples of corresponding solvates, including ethanol solvates, methanol solvates, etc. A "hydrate" is a solvate with water (H2O) molecules as solvent. One or more compounds of the present invention can be prepared as a solvate at will. 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, which was prepared using ethyl acetate and water. Similar methods for preparing solvates and hydrates are also described in EC van Tonder et al, AAPS Pharm Sci Tech., 5(1), article 12 (2004); and AL Bingham et al, Chem. Commun., 603-604 (2001). A typical, non-limiting preparation process is to dissolve the compound of the invention in a desired amount of a desired solvent (organic solvent or water or a mixture thereof) at a temperature above room temperature, cool the solution, allow the solution to crystallize, and then separate and select the crystals using standard methods. IR spectroscopy can be used to confirm the presence of the solvent (water) that forms the solvate (hydrate) in the crystals.

[0042] The term "active metabolite" refers to an active derivative of a compound that is formed when the compound is metabolized.

[0043] The term "polymorphs" refers to compounds of the invention that exist in different crystal lattice forms.

[0044] The term "isotopically labeled" refers to a compound of the present invention that is isotopically labeled. For example, the isotopes in the compound of the present invention may include various isotopes of elements such as H, C, N, O, P, F, and S, such as 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.

[0045] 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 invention that, upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the invention or a pharmaceutically active metabolite or residue thereof. Particularly preferred derivatives or prodrugs are those that, when administered to a patient, can increase the bioavailability of the compound of the invention (e.g., by making an orally administered compound more readily absorbed into the blood), or those that promote the delivery of the parent compound to biological organs or sites of action (e.g., the brain or lymphatic system). Prodrugs can be prepared by modifying functional groups present in the compound in a manner that allows for decomposition to the parent compound, either by conventional manipulation or in vivo. Various prodrug forms are well known in the art. See, for a discussion of prodrugs, T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) Vol. 14 of the ACSSymposium Series, Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and in Pergamon Press. Design of Prodrugs, Bundgaard, A. Ed., Elseview, 1985 and Method in Enzymology, Widder, K. et al., Ed.; Academic, 1985, vol. 42, p. 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, the above documents are incorporated herein by reference.

[0046] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule. The compounds of the present invention contain structures such as asymmetric or chiral centers, double bonds, etc. Therefore, the compounds of the present invention may include multiple isomeric forms such as optical isomers, geometric isomers, tautomers, atropisomers, etc. These isomers and their single isomers, racemates, etc. are all included within the scope of the present invention. For example, for optical isomers, optically active (R)- and (S)-isomers as well as D and L isomers can be prepared by chiral resolution, chiral synthesis or chiral reagents or other conventional techniques. For example, diastereomers can be converted into diastereomers by reaction with an appropriate optically active substance (such as a chiral alcohol or Mosher's acyl chloride), which can be separated and converted (such as hydrolyzed) into the corresponding single isomers. For another example, separation can also be performed by chromatographic column.

[0047] The "pharmaceutical compositions" herein can be prepared in a manner well known in the pharmaceutical art and can be administered or applied by a variety of routes, depending on whether local or systemic treatment is required and the area to be treated. Administration can be topical (e.g., transdermal, skin, eye and mucous membranes including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, such as intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus, or can be administered, for example, by a continuous infusion pump. The pharmaceutical compositions herein include, but are not limited to, the following forms: tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or dissolved in a liquid vehicle); ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0048] The pharmaceutical compositions herein can be formulated in unit dosage form, each dose containing about 0.1 to 1000 mg, usually about 5 to 1000 mg, more usually about 100 to 500 mg of active ingredient. The term "unit dosage form" refers to physically discrete units suitable as single dosage units for human patients and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in admixture with a suitable pharmaceutical carrier.

[0049] The term "subject" refers to an individual suffering from a disease, disorder, condition, etc., including mammals and non-mammals. 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); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, etc.

[0050] The term "treat" and other similar synonyms include alleviating, reducing or ameliorating the symptoms of a disease or condition, preventing other symptoms, ameliorating or preventing the underlying metabolic causes of symptoms, inhibiting the disease or condition, such as preventing the development of the disease or condition, alleviating the disease or condition, making the disease or condition better, alleviating the symptoms caused by the disease or condition, or stopping the symptoms of the disease or condition. In addition, the term may also include the purpose of prevention. The term also includes obtaining a therapeutic effect and / or a prophylactic effect. The therapeutic effect refers to curing or improving the underlying disease being treated. In addition, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, for example, although the patient may still be affected by the underlying disease, the patient's condition is observed to improve. In terms of prophylactic effect, the composition or compound can be administered to a patient at risk for a particular disease, or even if a diagnosis of the disease has not yet been made, the composition or compound can be administered to a patient who has one or more physiological symptoms of the disease.

[0051] The term "amount to achieve the necessary therapeutic effect" or "therapeutically effective amount" refers to the amount of at least one pharmaceutical agent or compound that, after administration, is sufficient to relieve to some extent one or more symptoms of the disease or condition being treated. The result can be a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other desired change in a biological system. Techniques such as dose escalation studies can be used to determine the effective amount appropriate for any individual case. The actual amount of compound, pharmaceutical composition, or medicament administered is generally determined by the physician based on relevant circumstances, including the condition being treated, the route of administration chosen, the actual compound administered; the age, weight, and response of the individual patient; the severity of the patient's symptoms, etc.

[0052] The proportion or concentration of the compound of the invention in the pharmaceutical composition may not be fixed and depends on a variety of factors, including dosage, chemical properties (e.g., hydrophobicity), route of administration, etc. For example, the compound of the invention can be provided in a physiologically buffered aqueous solution containing about 0.1 to 10% w / v of the compound for parenteral administration. Some typical dosage ranges are from about 1 μg / kg to about 1 g / kg body weight / day. In certain embodiments, the dosage range is from about 0.01 mg / kg to about 100 mg / kg body weight / day. The dosage is likely to depend on such variables as the type and extent of the disease or condition, the general health of the particular patient, the relative biological efficacy of the selected compound, the excipient formulation, and its route of administration.

[0053] The term "administering" refers to a method for delivering a compound or composition to a desired site for a biological effect. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, 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, such as those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.

[0054] The term "IC 50 ” refers to a 50% inhibition of the maximal effect in the assay measuring such effect.

[0055] Compound

[0056] In one aspect, the present application provides an azacycloalkane compound represented by Formula I or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer, or prodrug thereof:

[0057] Wherein, R1 is -NHR 11 、-OR 11 、-SR 11 or -C(=O)NHR 11 ; Among them, R 11 are each independently selected from 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 Heterocyclic group, 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 Heteroaryl acyl, or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl, thiol, substituted C 3～8 Cycloalkyl, substituted C 3～8 Heterocyclic, substituted C 6～20 Aryl or substituted C 5～20 heteroaryl;

[0058] R2 is selected from 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, carboxyl or mercapto; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0059] R3 is selected from cyano, -CONH2 or carboxyl;

[0060] L is selected from -S(=O)2-, -C(=O)-, -CH2-, or -S(=O)(=N)R L -, where R L Selected from hydrogen or substituted or unsubstituted C 1～8 Alkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0061] R4 is selected from halogen, substituted or unsubstituted C 1～8 Alkyl, -NR 31 R32 , 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, hydrazide, C 1～8 Alkylsulfonyl, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, -COR 33 , carboxyl or thiol;

[0062] A1 is selected from C atoms;

[0063] W1, W2, W3, W4 and W5 are each independently selected from C atoms or N atoms, and one, two or three of W1, W2, W3, W4 and W5 are N atoms; the dotted circle represents the bond forming the aromatic ring;

[0064] R 21 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0065] R 22 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0066] R 23 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0067] R 24 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0068] R 25 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0069] Or, R 24 、R 24 The connected W4 and R 25 and R 25 The attached W5, taken together, forms a 5-membered aryl, cycloalkyl, heteroaryl or heterocycloalkyl ring fused to the 6-membered ring formed by A1, W1, W2, W3, W4 and W5;

[0070] R 31 and R 32 are each independently selected from hydrogen, sulfonamide, 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, -C(=O)OR 34 or sulfhydryl;

[0071] R 33 is selected from 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0072] R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, carboxyl or thiol;

[0073] n is 1, 2, or 3;

[0074] Furthermore, the compound is not the following compound:

[0075] The present inventors have found through research that the compound of the present invention has good Rho kinase inhibitory activity, can be used as an inhibitor targeting ROCK, and can be used to regulate Rho kinase-mediated diseases.

[0076] In one embodiment, R1 is selected from -NHR 11 , where R 11 are each independently selected from 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 Heterocyclic group, 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 Heteroaryl acyl, or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl, thiol, substituted C 3～8 Cycloalkyl, substituted C 3～8 Heterocyclic, substituted C 6～20 Aryl or substituted C 5～20 heteroaryl;

[0077] Among them, R 31 and R 32 are each independently selected from 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, -C(=O)OR 34 or sulfhydryl;

[0078] R 33 is selected from 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR31 R 32 , hydroxyl, carboxyl or thiol;

[0079] R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, carboxyl or mercapto.

[0080] In one embodiment, R1 is selected from -NHR 11 , where R 11 is selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl, substituent selected from -NR 31 R 32 、-C(=O)R 33 or substituted or unsubstituted C 3～8 Heterocyclic group. For example, R 11 can be selected from hydrogen, optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted isopropyl, optionally substituted butyl, optionally substituted tert-butyl, optionally substituted pentyl, optionally substituted hexyl, etc., and the substituent can be selected from -NR 31 R 32 、-C(=O)R 33 or substituted or unsubstituted C 3～8 Heterocyclic group (substituents may be C 1～6 alkyl, such as methyl, ethyl, propyl, isopropyl, etc.) etc. Preferably, R 11 Selected from substituted or unsubstituted C 1～6 Alkyl, substituent selected from -NR 31 R 32 , where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 3～8 Heterocyclic group (substituents may be C 1～6 alkyl, such as methyl, ethyl, propyl, isopropyl, etc.). Preferably, R 11 Selected from substituted or unsubstituted C 1～6 Alkyl, substituent selected from -C(=O)R33 , where R 33 are independently substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 3～8 Heterocyclic group (substituents may be C 1～6 alkyl, such as methyl, ethyl, propyl, isopropyl, etc.).

[0081] In one embodiment, R1 is selected from -NH2, -NHC(=O)CH3, or

[0082] In one embodiment, R1 is selected from -OR 11 , where R 11 are each independently selected from 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 Heterocyclic group, 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 Heteroaryl acyl, or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl or thiol;

[0083] Preferably, R 11 Selected from hydrogen or substituted or unsubstituted C 1～6 Alkyl, substituent selected from -NR 31 R 32 、-C(=O)R 33 or substituted or unsubstituted C 3～8 heterocyclic group;

[0084] Among them, R 31 and R 32are each independently selected from 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, -C(=O)OR 34 or sulfhydryl;

[0085] R 33 is selected from 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0086] R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C5～20 heteroaryl, cyano, hydroxy, carboxyl or mercapto.

[0087] In one embodiment, R1 is selected from -OR 11 , where R 11 is hydrogen or C 1-6 Alkyl (e.g. C 1-3 alkyl groups such as methyl, ethyl, propyl, isopropyl).

[0088] In one embodiment, R1 is selected from -C(=O)NHR 11 , where R 11 are each independently selected from 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 Heterocyclic group, 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 Heteroaryl acyl, or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl or thiol;

[0089] Preferably, R 11 Selected from hydrogen or substituted or unsubstituted C 1～6 Alkyl, substituent selected from -NR 31 R 32 、-C(=O)R 33 or substituted or unsubstituted C 3～8 heterocyclic group;

[0090] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C2-6 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl, substituted or unsubstituted C 3～8 Cycloalkyl, substituted or unsubstituted C 3～8 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, -C(=O)OR 34 or sulfhydryl;

[0091] R 33 is selected from 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol;

[0092] R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, carboxyl or mercapto.

[0093] In one embodiment, R1 is selected from -C(=O)NHR 11 , where R 11 is hydrogen or C 1-6 Alkyl (e.g. C 1-3 alkyl groups such as methyl, ethyl, propyl, isopropyl).

[0094] In the above embodiments regarding R1, R 31 and R 32 can be independently hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 3～8 Heterocyclic group (substituents may be C 1～6 alkyl, such as methyl, ethyl, propyl, isopropyl, etc.). R 33 may be independently substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 3～8 Heterocyclic group (substituents may be C 1～6 alkyl, such as methyl, ethyl, propyl, isopropyl, etc.).

[0095] In one embodiment, R2 is selected from hydrogen and R3 is selected from cyano.

[0096] The dotted circle in Formula I represents the bond forming the aromatic ring. In one embodiment, W1, W2, W3, W4 and W5 are each independently selected from a C atom or a 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 a N atom, and one or two of W1, W2, W4 and W5 are N atoms.

[0097] In one embodiment,

[0098] In formula I Can be selected from the following A1-A7 structures:

[0099] or

[0100] Wherein, in structures A1-A7, R7 are each independently selected from hydrogen, halogen, -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0101] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C1～6 Alkyl, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl;

[0102] Wherein, the substituent is selected from halogen, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxy, carboxyl or mercapto.

[0103] In the above structural formula, * indicates the position of connection to the pyrazole ring structure.

[0104] In one embodiment, each R7 is independently selected from hydrogen, -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0105] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 5～20 Heteroaryl or substituted or unsubstituted C 3～8 heterocyclic group;

[0106] Wherein, the substituent is selected from C 1～8 alkyl.

[0107] In one embodiment, in Formula I The following A1 structures can be used:

[0108] Among them, in the A1 structure, R7 is hydrogen.

[0109] In one embodiment, in Formula I For the following A2 structure:

[0110] Among them, in the A2 structure, R7 is -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0111] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl;

[0112] Wherein, the substituent is selected from C 1～8 Alkyl or hydroxyl.

[0113] In one embodiment, in Formula I For the following A3 structure:

[0114] Among them, in the A3 structure, R7 is -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0115] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl;

[0116] Wherein, the substituent is selected from C 1～8 Alkyl or hydroxyl.

[0117] In one embodiment, in Formula I For the following A4 structure:

[0118] Among them, in the A4 structure, R7 is -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0119] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl;

[0120] Wherein, the substituent is selected from C 1～8 Alkyl or hydroxyl.

[0121] In one embodiment, in Formula I For the following A5 structure:

[0122] Among them, in the A5 structure, R7 is -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0123] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C5～20 heteroaryl;

[0124] Wherein, the substituent is selected from C 1～8 Alkyl or hydroxyl.

[0125] In one embodiment, in Formula I For the following A6 structure:

[0126] Among them, in the A6 structure, R7 is -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0127] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl;

[0128] Wherein, the substituent is selected from C 1～8 Alkyl or hydroxyl.

[0129] In one embodiment, in Formula I For the following A7 structure:

[0130] Among them, in the A7 structure, R7 is -NR 31 R 32 or substituted or unsubstituted C 5～20 heteroaryl,

[0131] Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl;

[0132] Wherein, the substituent is selected from C 1～8 Alkyl or hydroxyl.

[0133] In the above embodiment, R7 can be -NHCH3, -NHCH2CH2OH, Cl,

[0134] or

[0135] In one embodiment, L is selected from -S(=O)2-, -C(=O)-, or -S(=O)(=N)R L -, where R L Selected from hydrogen or C 1～6In one embodiment, L is selected from -S(=O)2-. In one embodiment, L is selected from -C(=O)-. In one embodiment, L is selected from -S(=O)(=N)R L -, where R L Selected from hydrogen or C 1～6 Alkyl groups, such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, pentyl or hexyl, etc.

[0136] In one embodiment, R4 is selected from 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, hydrazide, C 1～8 Alkylsulfonyl, C 1～8 Alkyl, C 1～8 Halogenated alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, -COR 33 , carboxyl or thiol;

[0137] Among them, R 31 and R 32 are each independently selected from hydrogen, sulfonamide, 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, cyano, -NR 31 R 32 , hydroxyl, -C(=O)OR 34 or sulfhydryl.

[0138] In one embodiment, R4 is selected from substituted or unsubstituted C 1～8Alkyl (such as methyl, ethyl, propyl, etc.), wherein the substituent can be halogen, hydroxyl, cyano, C 1～8 Alkylsulfonyl, etc., for example, R4 can be methyl, ethyl, trifluoroethyl, trifluoromethyl, methylsulfonylmethyl, etc.

[0139] In one embodiment, R4 is selected from substituted or unsubstituted C 3～8 Cycloalkyl (e.g., cyclopropyl, cyclobutyl, etc.), wherein the substituent may be halogen, hydroxyl, cyano, etc., for example, R4 may be 2,2-difluorocyclopropyl, 2-cyanocyclopropyl, etc.

[0140] In one embodiment, R4 is selected from -NR 31 R 32 , where R 31 and R 32 are each independently selected from hydrogen, sulfonamide, 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 Heterocyclic group, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, cyano, -NR 31 R 32 , hydroxyl, -C(=O)OR 33 or thiol; R 33 Selected from hydrogen or C 1～6 Alkyl (eg, methyl, ethyl, propyl, etc.). In one embodiment, R4 is selected from -NR 31 R 32 , where R 31 is hydrogen, R 32 Selected from substituted or unsubstituted C 1～8 Alkyl (such as methyl, ethyl, propyl, etc.), substituted or unsubstituted C 3～8 Cycloalkyl (e.g. cyclopropyl), or substituted or unsubstituted C 5～20 Heteroaryl (e.g., thiadiazolyl, etc.), wherein the substituent may be halogen, hydroxyl, cyano, etc. In one embodiment, R4 is selected from -NR 31 R 32 , where R 31 is hydrogen, R 32 Independently selected from substituted or unsubstituted C 1～6 Alkyl (such as methyl, ethyl, propyl, etc.), substituent selected from -C(=O)OR 34 , where R34 Selected from hydrogen or C 1～6 Alkyl (eg, methyl, ethyl, propyl, etc.).

[0141] 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; when n is 3, the azacycloalkane ring is azacyclohexyl.

[0142] In one embodiment, the azacycloalkane compound is selected from the following compounds:

[0143] In a preferred embodiment, the azacycloalkane compound is selected from the following compounds:

[0144] The compounds described herein can be prepared by the following methods. The following methods and examples are intended to illustrate these methods. These schemes and examples should not be construed as limiting the present invention in any way. The compounds described herein can also be synthesized using standard synthetic techniques known to those skilled in the art, or a combination of methods known in the art and the methods described herein.

[0145] The chemical reactions described in the examples of the present invention are carried out in suitable solvents that are compatible with the chemical transformations of the present invention and the reagents and materials required. To obtain the compounds of the present invention, it may sometimes be necessary for those skilled in the art to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0146] An important consideration in planning any synthetic route in this field is the selection of an appropriate protecting group for a reactive functional group, such as the amino group in the present invention. For the trained practitioner, Greene and Wuts (Protective Groups In Organic Synthesis, Wiley and Sons, 1991) is an authority on this subject. All references cited herein are incorporated herein in their entirety.

[0147] The reactions described herein can be monitored by any suitable method known in the art. For example, the reactions can be monitored by broad spectrum methods such as nuclear magnetic resonance spectroscopy (e.g. 1 H or 13C), infrared spectroscopy, spectrophotometry (eg, UV-visible), mass spectrometry, or the like, or by chromatography, such as high performance liquid chromatography (HPLC) or thin layer chromatography, to monitor product formation.

[0148] The compounds of the general formula I of the present invention can be prepared by those skilled in the art of organic synthesis using the following schemes and standard methods in the art.

[0149] Process 1:

[0150] Compound a1 and compound a2 undergo Michael addition reaction to generate compound a3, compound a3 is deprotected to generate compound a4, and compound a4 undergoes substitution reaction with compound a5 to generate the compound of formula I of the present invention.

[0151] Process 2:

[0152] Compound b1 and compound b2 undergo a coupling reaction to generate compound b3, compound b3 reacts with a substituted boronic acid or a substituted amino compound to generate compound b4, compound b4 is deprotected to generate compound b5, and compound b5 and compound b6 undergo a Michael addition reaction to obtain the compound of formula I of the present invention.

[0153] Pharmaceutical compositions and uses

[0154] The present invention also provides a pharmaceutical composition comprising the compound described in any one of the above technical solutions or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and a pharmaceutically acceptable carrier.

[0155] The pharmaceutical composition includes, but is not limited to, oral dosage forms, parenteral dosage forms, external dosage forms, rectal dosage forms, etc. For example, the pharmaceutical composition can be oral tablets, capsules, pills, powders, sustained-release preparations, solutions and suspensions, sterile solutions, suspensions or emulsions for parenteral injection, ointments, creams, gels, etc. for external use, eye drops for external use, inhalants for external use, or suppositories for rectal administration.

[0156] The pharmaceutical composition may further include other active ingredients or drugs, which are used in combination with the compound or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope label, isomer or prodrug.

[0157] The present invention also provides the use of the above-mentioned compound or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled form, isomer or prodrug, and the above-mentioned pharmaceutical composition in the preparation of a medicament for treating Rho kinase-mediated diseases. Rho kinase may include types such as ROCK1 and ROCK2.

[0158] The present application also relates to a method for treating a disease mediated by Rho kinase, comprising administering a therapeutically effective amount of the aforementioned azacycloalkane compound, or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled compound, isomer, or prodrug thereof, or the aforementioned pharmaceutical composition, to a patient in need thereof. Rho kinases may include types such as ROCK1 and ROCK2.

[0159] In one embodiment, the Rho kinase-mediated disease is asthma, cancer, glaucoma, insulin resistance, renal failure, neuronal degeneration, or osteoporosis.

[0160] The azacycloalkane compound and the pharmaceutical composition thereof provided by the present invention have significant Rho kinase inhibitory activity, and their enzymatic activity and cell activity are superior to those of existing ROCK inhibitors such as Ripasudil, Netarsudil and belumosudil, and therefore have great application potential.

[0161] In order to make the objectives, technical solutions and advantages of the present invention more clear, the technical solutions of exemplary embodiments of the present invention will be further described below.

[0162] Example 1:

[0163] 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

[0164] Synthesis route:

[0165] 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

[0166] 46 g (0.1 mol, 1.0 eq) of 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) of tert-butyl 3-(cyanomethyl)azetidine-1-carboxylate were dissolved in 200 mL of N,N-dimethylformamide. 1.5 g (0.01 mol, 0.1 eq) of 1,8-diazabicyclo[5.4.0]undec-7-ene was slowly added dropwise to the reaction system at room temperature. The reaction was stirred at room temperature overnight. The reaction was quenched with water, and the aqueous phase was extracted with ethyl acetate and washed twice with water. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to obtain the product (61 g, yield = 93%).

[0167] 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

[0168] 61 g (93 mmol, 1.0 eq) of 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 of dichloromethane. 50 mL of 4N hydrochloric acid in dioxane was added at room temperature and the mixture was stirred overnight. The reaction was monitored by TLC. The reaction solution was evaporated to dryness to obtain the product (55 g, yield = 100%).

[0169] 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

[0170] Under ice, 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). Triethylamine (3.2 g, 32.46 mmol, 6.0 eq) was slowly added dropwise to the reaction solution and stirred for 15 min. 2,2,2-Trifluoroethane-1-sulfonyl chloride (987 mg, 5.41 mmol, 1.0 eq) dissolved in 5 mL of dichloromethane was then added dropwise to the reaction solution. The mixture was allowed to react at 25°C for 2 h. After completion of the reaction, the reaction solution was evaporated to dryness under reduced pressure, and the reaction solution was concentrated and the residue was purified by column chromatography (PE:EA=1:2) to give 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.

[0171] LC-MS (ESI), m / z: [M+H] + =700.9.

[0172] Step D: 2-(3-(3-(1,3-dioxoisoindol-2-yl)-4-(7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-1-((2,2,2-trifluoroethyl)sulfonyl)azetidin-3-yl)acetonitrile

[0173] Dissolve 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 (700 mg, 1.0 mmol, 1.0 eq) in acetonitrile (6 mL). Add boron trifluoride-ether solution (2 mL) dropwise in an ice bath. Stir the reaction mixture at 25°C for 2 h. After completion of the reaction, evaporate to dryness under reduced pressure. The crude product, 2-(3-amino-4-hydroxymethyl-7H-pyrrolyl)pyrimidin-4-yl-1-pyrazol-1-(2,2,2-trifluoroethylsulfonyl)azetidin-3-ylacetonitrile (450 mg, crude), was obtained as a yellow oil.

[0174] LC-MS (ESI), m / z: [M+H] + =601.

[0175] 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

[0176] 2-(3-(3-(1,3-dioxoisoindol-2-yl)-4-(7-(hydroxymethyl)-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 under ice-cooling. After completion of the reaction, water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL*3). The combined organic phases were 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 um, mobile phase: ACN-H2O (0.1% FA), gradient 20%-50%, flow rate: 20 mL / min) to give the product (45 mg, yield = 11%).

[0177] LC-MS (ESI), m / z: [M+H] + =440.9.

[0178] 1 H-NMR (400MHz, D2O) δ8.61(s,1H),8.29(s,1H),7.51(d,J=3.6Hz,1H),6.76(d,J=3.6Hz,1H),4.33(d,J=9.3Hz,2H),3.88(s,4H),3.44(s,2H).

[0179] The following examples were prepared with reference to the experimental route and method in Example 1:

[0180] Example 10: 3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)-3-(cyanomethyl)-N-methylazetidine-1-carboxamide

[0181] Synthesis route:

[0182] Step A: 2-{3-[3-(1,3-dioxoisoindole)-4-[7-(hydroxymethyl)pyrrolidin[2,3-d]pyrimidine]pyrazole]3-azetidine}acetonitrile

[0183] 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. After complete dissolution, trifluoroacetic acid (100 mL) was added and the mixture was stirred in an oil bath at 30°C overnight. The product, 2-{3-[3-(1,3-dioxoisoindol)-4-[7-(hydroxymethyl)pyrrolidino[2,3-d]pyrimidinyl]pyrazole]3-azetidine}acetonitrile (12.0 g, crude) was obtained after spin drying.

[0184] LC-MS (ESI), m / z: [M+H] + =455.0.

[0185] Step B: 2-[3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)azetidin-3-yl]acetonitrile

[0186] 2-{3-[3-(1,3-dioxoisoindole)-4-[7-(hydroxymethyl)pyrrolidino[2,3-d]pyrimidinyl]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 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 x 5). The organic phase was collected, dried, concentrated under reduced pressure, and the residue was purified by C18 column to obtain 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%).

[0187] LC-MS (ESI), m / z: [M+H] + =295.0.

[0188] Step C: Phenyl N-methylcarbamate

[0189] To a solution of methylamine (2 g, 0.03 mol) in tetrahydrofuran (15 mL) at 25°C was added sodium bicarbonate (2.77 g, 0.03 mol). The reaction mixture was stirred at 25°C for 15 minutes. Phenyl chloroformate (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 x 3). The organic phase was dried and concentrated. The residue was purified by column chromatography (DCM:MeOH = 20:1) to afford the product (1 g, 22%).

[0190] LC-MS (ESI), m / z: [M+H] + =152.1.

[0191] Step D: 3-(3-amino-4-{7H-pyrrolo[2,3-d]pyrimidin-4-yl}pyrazol-1-yl)-3-(cyanomethyl)-N-methylazetidine-1-carboxamide

[0192] 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 then added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction solution 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%).

[0193] LC-MS (ESI), m / z: [M+H] + =352.0.

[0194] 1 H-NMR (400 MHz, 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). The following examples were prepared with reference to the experimental route and method in Example 10:

[0195] Example 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

[0196] Synthesis route:

[0197] 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

[0198] To a solution of 2-cyanoacetic acid (92 mg, 1.08 mmol, 2.0 eq), triethylamine (220 mg, 2.2 mmol, 4.0 eq) in dichloromethane (5 mL) was added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (410.4 mg, 1.08 mmol, 2.0 eq), and the reaction solution was stirred at 25 ° C for 0.5 h. 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 to the reaction solution, and the reaction solution was stirred at 25 ° C for 2 h. The reaction mixture was diluted with water (20 mL) and then extracted with dichloromethane (20 mL*3). The combined organic phases were washed with saturated brine (20 mL), 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 afford the product 3-(3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(hydroxymethyl)-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.

[0199] LC-MS (ESI), m / z: [M+H] + =622.0.

[0200] Step B: 3-(3-(cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl))-1H-pyrazol-1-yl)azetidin-1-yl)-3-oxopropionitrile

[0201] 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 etherate (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 x 3). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the crude product (140 mg, 76% yield).

[0202] LC-MS (ESI), m / z: [M+H] + =522.0.

[0203] 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

[0204] 3-(3-(Cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(hydroxymethyl)-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. Ethylenediamine (0.5 mL) was added under ice-cooling to adjust the pH to 8-9. The reaction system was stirred at 25°C for 1 h. The reaction solution was concentrated and analyzed by Prep-HPLC (column model: Gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% TFA), gradient 25%-40%, flow rate: 20 mL / min) to obtain the product (31.9 mg, 33%).

[0205] LC-MS (ESI), m / z: [M+H] + =362.0.

[0206] 1H-NMR (400MHz, DMSO-d6) δ8.63(d,J=9.9Hz,1H),8.57(d,J=7.4Hz,1H),7.54(d,J=5.3Hz,1H),7.03(d,J=9.7Hz,1H),4.67(d ,J=12.9Hz,1H),4.52(d,J=10.6Hz,1H),4.38(d,J=10.6Hz,1H),4.14(d,J=12.1Hz,1H),3.78(s,2H),3.56(d,J=9.3Hz,2H).

[0207] The following examples were prepared with reference to the experimental route and method in Example 21:

[0208] Example 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

[0209] Synthesis route:

[0210] Step A: N-(tert-Butyldimethylsilyl)methanesulfonamide

[0211] Under nitrogen protection, methylsulfonamide (1g, 1.53mmol, 1.0eq) was dissolved in tetrahydrofuran (15mL), triethylamine (464mg, 4.59mmol, 3.0eq) was added at room temperature, and a toluene (50mL) solution of tert-butyldimethylsilyl chloride (462mg, 3.06mmol, 2.0eq) was added dropwise. The reaction was allowed to proceed overnight at room temperature, filtered, and the solid was washed with ether (200ml). Ether (20mL) was added to the filtrate, allowed to stand for 30min, and filtered again. The filtrate was concentrated to obtain a crude product. The crude product was purified by Combiflash column (petroleum ether: ethyl acetate = 1:1) to obtain the product N-(tert-butyldimethylsilyl) methanesulfonamide (1.2g, 39% yield) as a white solid.

[0212] LC-MS (ESI), m / z: [M+H] + =210.0.

[0213] 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

[0214] Under nitrogen protection, 0.36M 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 stirred for 15 min. Then, N-(tert-butyldimethylsilyl)methanesulfonamide (1.13 g, 5.4 mmol, 6.0 eq) was added at 0 ° C and stirred for 20 min. The reaction solution was added dropwise to 2-(3-(3-(1,3-dioxoisoindolin-2-yl))-4- To a solution of (7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (500 mg, 0.90 mmol, 1.0 eq) in dichloromethane (2 mL) was added dropwise. The mixture was then warmed to room temperature and stirred for 2 h. Water was added to quench the reaction, followed by extraction with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The crude product was purified via a Combiflash column (dichloromethane:methanol = 97:3) to obtain the crude product as a yellow solid (1.1 g, purity: 50%, yield: 82%).

[0215] LC-MS (ESI), m / z: [M+H] + =746.0.

[0216] Step D: 2-(3-(3-(1,3-dioxoisoindolin-2-yl))-4-(7-(hydroxymethyl)-7-hydro-pyrrolo[2,3-d]pyrimidin-4-yl)-1-hydro-pyrazol-1-yl)-1-(S-methylsulfonylimino)azetidin-3-yl)acetonitrile

[0217] At room temperature, 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), and boron trifluoride etherate (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 phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to give the crude product (500 mg, crude product).

[0218] LC-MS (ESI), m / z: [M+H] + =531.9.

[0219] 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

[0220] At room temperature, 2-(3-(3-(1,3-dioxoisoindolin-2-yl))-4-(7-(hydroxymethyl)-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) and ethylenediamine (0.5 mL) was added under ice bath to adjust the pH to 8-9. The reaction system was stirred at 25°C for 1 h. The reaction solution was concentrated and the product was obtained by Prep-HPLC (column model: gemini-C18 150x 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% TFA), gradient 30%-35%, flow rate: 20 mL / min) to obtain the product (76 mg, 26%).

[0221] LC-MS (ESI), m / z: [M+H] + =372.0.

[0222] 1H-NMR (400MHz, DMSO-d6) δ11.96(d,1H),8.66(s,1H),8.62(s,1H),7.56(d,J=3.6Hz,1H),7.12(d,J=3.6Hz,1 H), 6.40 (s, 2H), 4.43 (dd, J = 11.7, 9.3Hz, 2H), 4.03 (d, J = 9.6Hz, 2H), 3.92 (s, 1H), 3.53 (s, 2H), 2.96 (s, 3H).

[0223] The following examples were prepared with reference to the experimental route and method in Example 36:

[0224] 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

[0225] Synthesis route:

[0226] Step A: tert-Butyl (chlorosulfonyl)carbamate

[0227] 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 0°C under nitrogen. The resulting reaction mixture was stirred at 25°C for 20 minutes. After completion of the reaction, the mixture was concentrated to half its original volume, protected with nitrogen, and refrigerated for 50 minutes. Then, n-hexane (10 mL) was added, the reaction mixture was filtered, and the precipitate was washed with n-hexane. The filter cake was collected and dried to obtain tert-butyl (chlorosulfonyl)carbamate (2 g, crude product), which was used directly in the next step.

[0228] 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

[0229] At 0 ° C, 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 added to dichloromethane (20 mL) and dissolved. After that, 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, 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 by silica gel column chromatography (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%).

[0230] LC-MS (ESI), m / z: [M+H] + =733.9.

[0231] 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

[0232] 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) were 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). The reaction solution was stirred at 25°C for 2 h. The reaction solution was diluted with water (20 mL) and then extracted with ethyl acetate (20 mL*3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by Combiflush column (petroleum ether:ethyl acetate = 3:1) to obtain 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.

[0233] LC-MS (ESI), m / z: [M+H] + =815.9.

[0234] Step D: 3-(Cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-N-(2,2,2-trifluoroethyl)azetidine-1-sulfonamide

[0235] 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 etherate (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%).

[0236] LC-MS (ESI), m / z: [M+H] + =615.9.

[0237] 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

[0238] 3-(Cyanomethyl)-3-(3-(1,3-dioxoisoindolin-2-yl)-4-(7-(hydroxymethyl)-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. Ethylenediamine (0.2 mL) was added under ice-cooling to adjust the pH to 8-9. The reaction system was stirred at 25°C for 1 h. The reaction solution was concentrated and analyzed by Prep-HPLC (column model: Gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% TFA), gradient 35%-40%, flow rate: 20 mL / min) to obtain the product (17.5 mg, yield = 55%).

[0239] LC-MS (ESI), m / z: [M+H] + =455.7.

[0240] 1 H-NMR (400MHz, CD3OD) δ8.58(s,1H),8.40(s,1H),7.34(d,J=3.6Hz,1H),6.86(d,J=3.6H z, 1H), 4.39 (d, J = 9.3Hz, 2H), 4.02 (d, J = 9.4Hz, 2H), 3.68 (q, J = 9.1Hz, 2H), 3.38 (s, 2H).

[0241] The following examples were prepared with reference to the experimental route and method in Example 40:

[0242] Example 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-carboxamide

[0243] Synthesis route:

[0244] Step A: Ethyl 2-cyano-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)acetate

[0245] 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 an additional 1 h. After the reaction, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (20 mL x 3). The organic phase was dried, concentrated, evaporated, and purified on a silica gel column (petroleum ether:ethyl acetate = 5:1) to obtain ethyl 2-cyano-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)acetate (1.1 g, yield = 88%).

[0246] LC-MS (ESI), m / z: [M+H] + =361.0.

[0247] Step B: Ethyl (2E)-3-amino-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)prop-2-enoate

[0248] 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), diisobutylaluminum hydride (1 M in THF, 8.0 mL) was added dropwise at 0 ° C., and the solution was stirred at 25 ° C. for 3 h. After the reaction, the mixture was diluted with water (50 mL) and extracted with ethyl acetate (30 mL*3). The organic phase was dried, concentrated and evaporated to dryness, and purified on a silica gel column (petroleum ether: ethyl acetate = 1:1) to give the product ethyl (2E)-3-amino-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)prop-2-enoate (700 mg, yield = 70.0%).

[0249] LC-MS (ESI), m / z: [M+H] + =363.1.

[0250] Step C: 4-{7-[(2-methoxyethyl)trimethyl-{5}-silanyl]pyrrolo[2,3-d]pyrimidin-4-yl}-1H-pyrazol-3-ol

[0251] (2E)-3-Amino-2-(7-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-d]pyrimidin-4-yl)prop-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 hours. The reaction solution was concentrated and evaporated to dryness, and then purified on a silica gel column (dichloromethane:methanol = 20:1) to obtain the product 4-{7-[(2-methoxyethyl)trimethyl-{5}-silanyl]pyrrolo[2,3-d]pyrimidin-4-yl}-1H-pyrazol-3-ol (600 mg, yield = 76%).

[0252] LC-MS (ESI), m / z: [M+H] + =332.1.

[0253] Step D: 3-cyanomethylene-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide

[0254] 2-(Azetidin-3-ylidene)acetonitrile hydrochloride (200 mg, 1.53 mmol, 1.0 eq) was dissolved in dichloromethane (8 ml), and N,N-diisopropylethylamine (593 mg, 4.59 mmol, 3.0 eq) was slowly added dropwise to the reaction mixture at 0°C. The reaction system was stirred at 25°C for 15 minutes, and then (2,2,2-trifluoroethyl)phenylcarbamate (337 mg, 1.53 mmol, 1.0 eq) was added to the reaction mixture. The reaction system was allowed to react at 25°C for 18 hours. After completion of the reaction, the reaction mixture was evaporated to dryness under reduced pressure. The residue was concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain 3-cyanomethylene-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide (200 mg, yield = 95%) as a white solid.

[0255] LC-MS (ESI), m / z: [M+H] + =262.0.

[0256] 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-carboxamide

[0257] 4-(2-Trimethylsilylethoxymethyl)pyrrolopyrrolidin-4-ylpyrazole (302 mg, 0.91 mmol, 1.0 eq) and 3-cyanomethylene-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide (200 mg, 0.91 mmol, 1.0 eq) were dissolved in N,N-dimethylformamide (8 mL). 1,8-diazabicyclo[5.4.0]undecene-7-ene (92 mg, 0.91 mmol, 1.0 eq) was then added to the reaction mixture. The reaction system was allowed to react at 25°C for 18 hours. After the reaction was completed, water (20 mL) was added to the reaction liquid, and the mixture was extracted with dichloromethane (20 mL * 3). The combined organic phases were washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The reaction solution was concentrated and the 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-carboxamide (100 mg, yield = 30%) as a yellow oil.

[0258] LC-MS (ESI), m / z: [M+H] + =551.0.

[0259] Step F: 3-cyanomethyl-3-(3-hydroxy-4-hydroxymethyl-7H-pyrrolylpyrrolyl)-2,3-d-pyrrolidin-4-yl-1-pyrazolyl-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide

[0260] 3-Cyanomethyl-3-hydroxy-4-(7-(2-trimethylsilylethoxymethyl)-7H-pyrrolidin-4-ylpyrimidin-1-pyrazol-1-yl-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide (150 mg, 0.27 mmol, 1.0 eq) was dissolved in dichloromethane (6 ml). Boron trifluoride-ether solution (2 mL) was added dropwise under ice-cooling. The reaction solution was stirred at 25° C. for 2 h. After completion of the reaction, the solution was evaporated to dryness under reduced pressure. The crude product, 3-cyanomethyl-3-(3-hydroxy-4-hydroxymethyl-7H-pyrrolylpyrrolyl)-2,3-d-pyrrolidin-4-yl-1-pyrazol-1-yl-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide (82 mg, crude) was obtained as a yellow oil.

[0261] LC-MS (ESI), m / z: [M+H] + =451.0.

[0262] Step D: 3-cyanomethyl-3-hydroxy-4-pyrrolidin-4-ylpyrimidine-2,3-dimethylpyrazol-1-yl-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide

[0263] 3-Cyanomethyl-3-(3-hydroxy-4-hydroxymethyl-7H-pyrrolylpyrrolyl)-2,3-d-pyrrolidin-4-yl-1-pyrazolyl-N-(2,2,2-trifluoroethyl)azetidine-1-carboxamide (82 mg, 0.18 mmol, 10.0 eq) was dissolved in methanol (5 mL) and ethylenediamine (0.5 mL) was added under ice-cooling. After the reaction was completed, water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL*3). The combined organic phases were 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 um, mobile phase: ACN-H2O (0.1% FA), gradient 10%-40%, flow rate: 20 mL / min) to give the product (5.3 mg, yield = 7%).

[0264] LC-MS (ESI), m / z: [M+H] + =421.0.

[0265] 1 H-NMR (400MHz, CD3OD) δ8.67(s,1H),8.60(s,1H),7.49(d,J=3.6Hz,1H),7.05(d,J=3.6H z, 1H), 4.56 (d, J = 9.3Hz, 2H), 4.28 (d, J = 9.3Hz, 2H), 3.82 (d, J = 9.3Hz, 2H), 3.48 (s, 2H).

[0266] The following examples were prepared with reference to the experimental route and method in Example 55:

[0267] Example 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-carboxamide

[0268] Synthesis route:

[0269] Step A: Methyl 4-bromo-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate

[0270] To a solution of methyl 4-bromo-1H-pyrazole-3-carboxylate (5 g, 25 mmol, 1.0 eq) in acetonitrile (50 mL) were 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 reaction mixture was concentrated under reduced pressure to afford the product, methyl 4-bromo-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate (5.88 g, crude product), as a yellow solid.

[0271] LC-MS (ESI), m / z: [M+H] + =325.0.

[0272] Step B: 1-(4-methoxybenzyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-3-carboxylic acid methyl ester

[0273] To a solution of methyl 4-bromo-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylate (5.88 g, 18 mmol, 1.0 eq), pinacol diboron (4.58 g, 18 mmol, 1.0 eq), and potassium acetate (5.292 g, 54 mmol, 3.0 eq) in 1,4-dioxane (200 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (1.316 g, 1.8 mmol, 0.1 eq). The reaction mixture was stirred at 90°C under nitrogen for 16 h. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The crude product was obtained and purified by Combiflash column (petroleum ether:ethyl acetate=2:1) ​​to obtain the crude product (3.74 g, crude product) in the form of a yellow solid.

[0274] LC-MS (ESI), m / z: [M+H] + =373.1.

[0275] Step C: Methyl 1-(4-methoxybenzyl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylate

[0276] 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 bistriphenylphosphine palladium dichloride (0.7 g, 1 mmol, 0.1 eq). The reaction mixture was stirred at 90°C under nitrogen for 16 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The crude product was obtained and purified by Combiflash column (dichloromethane:methanol=20:1) to obtain the crude product (1.85 g, crude product) in the form of a yellow solid.

[0277] LC-MS (ESI), m / z: [M+H]+=494.1.

[0278] Step D: Methyl 4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylate

[0279] To a solution of methyl 1-(4-methoxybenzyl)-4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylate (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 at 25°C overnight. The reaction mixture was diluted with water (20 mL) and then extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The crude product was purified by Combiflash column (dichloromethane:methanol = 15:1) to obtain the crude product as a yellow solid (900 mg, yield = 65%).

[0280] LC-MS (ESI), m / z: [M+H] + =374.1.

[0281] Step E: 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

[0282] To a solution of methyl 4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxylate (900 mg, 2.41 mmol, 1.0 eq) and 1,8-diazobisspiro[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). 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 x 3). The combined organic phases were washed with saturated brine (20 mL), 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 (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.

[0283] LC-MS (ESI), m / z: [M+H] + =568.2.

[0284] 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

[0285] 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 methanol (5 mL) with ammonia was stirred at 25° C. overnight. The mixture was evaporated to dryness under reduced pressure to give the crude product tert-butyl 3-(3-carbamoyl-4-(7-(2-(trimethylsilyl)ethoxy)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidin-1-carboxylate (195 mg, yield = 80%).

[0286] LC-MS (ESI), m / z: [M+H] + =553.2.

[0287] 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

[0288] A solution of tert-butyl 3-(3-carbamoyl-4-(7-(2-(trimethylsilyl)ethoxy)-7H-pyrrolo[2,3-d]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. The mixture was evaporated to dryness under reduced pressure to give the crude product 1-(3-(cyanomethyl)azepine-3-yl)-4-(7-(2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (135 mg, crude).

[0289] LC-MS (ESI), m / z: [M+H] + =453.1.

[0290] 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

[0291] To a solution of 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, 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 mixture was stirred at 25°C for 2 h. 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 x 3). The combined organic phases were washed with saturated brine (20 mL), 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 (dichloromethane:methanol=20:1) to give the product 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azepine-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.

[0292] LC-MS (ESI), m / z: [M+H] + =599.0.

[0293] Step I: 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azapyridin-3-yl)-4-(7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide

[0294] To a solution of 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, 0.17 mmol, 1.0 eq) in dichloromethane (5 mL) was added a boron trifluoride-ether solution (0.5 mL). The reaction solution was stirred at 25° C. for 2 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain the product 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azapyridin-3-yl)-4-(7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-3-carboxamide (63 mg, crude) as a yellow solid.

[0295] LC-MS (ESI), m / z: [M+H] + =499.0.

[0296] 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

[0297] To a solution of 1-(3-(cyanomethyl)-1-((2,2,2-trifluoroethyl)sulfonyl)azepine-3-yl)-4-(7-(hydroxymethyl)-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 under vacuum and stirred at 25°C for 2 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine (20 mL), 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 purified by Prep-HPLC (column model: gemini-C18 150 x 21.2 mm, 5 μm, mobile phase: ACN-H2O (0.1% TFA), gradient 25%-30%, flow rate: 20 mL / min) to give the product (3.2 mg, yield = 5.2%).

[0298] LC-MS (ESI), m / z: [M+H] + =469.0.

[0299] 1 H-NMR(400MHz,D2O)δ8.72(s,1H),8.69(s,1H),8.30(s,1H),7.57(d,J=3.6Hz,1H),6.74(d,J =3.5Hz, 1H), 4.77 (d, J = 9.4Hz, 2H), 4.46 (d, J = 9.3Hz, 2H), 4.39 (q, J = 9.2Hz, 2H), 3.60 (s, 2H).

[0300] The following examples were prepared with reference to the experimental route and method in Example 60:

[0301] Example 69: 2-[3-(3-amino-4-{6-[(1-methylpyrazol-4-yl)amino]pyrimidin-4-yl}pyrazol-1-yl)-1-methanesulfonylpropidin-3-yl]acetonitrile

[0302] Synthesis route:

[0303] Step A: tert-Butyl [3-amino-4-(2-chloropyrimidin-4-yl)pyrazol-1-yl]carboxylate

[0304] To a solution of tert-butyl [3-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]carboxylate (418 mg, 2.85 mmol, 1.0 eq) in 1,4-dioxane (10 mL) were added 2,4-dichloropyrimidine (644.44 mg, 4.28 mmol, 1.5 eq), sodium carbonate (611 mg, 5.82 mmol, 2.0 eq) and bistriphenylphosphine palladium dichloride (141.81 mg, After the reaction was completed, the reaction solution was evaporated to dryness and purified by silica gel column purification (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%).

[0305] LC-MS (ESI), m / z: [M+H] + =296.1.

[0306] Step B: 6-(3-amino-1H-pyrazol-4-yl)-N-(1-methylpyrazol-4-yl)pyrimidin-4-amine

[0307] 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 added to n-butanol (5 mL) at room temperature. After dissolution, p-toluenesulfonic acid (170 mg, 0.99 mmol, 3.0 eq) was added and stirred at 110°C for 4 hours. The mixture was then concentrated and evaporated to dryness. Purification was performed on a silica gel column (dichloromethane:methanol = 10:1) to obtain the product 6-(3-amino-1H-pyrazol-4-yl)-N-(1-methylpyrazol-4-yl)pyrimidin-4-amine (70 mg, yield = 80%).

[0308] LC-MS (ESI), m / z: [M+H] + =257.0.

[0309] Step C: 2-[3-(3-amino-4-{6-[(1-methylpyrazol-4-yl)amino]pyrimidin-4-yl}pyrazol-1-yl)-1-methanesulfonylpropidin-3-yl]acetonitrile

[0310] 6-(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 aziridine-3-ylidene)acetonitrile (70.5 mg, 0.41 mmol, 1.5 eq) were added to acetonitrile (5 mL), dissolved, and then 1,8-diazabicyclo[5.4.0]undec-7-ene (41.04 mg, 0.27 mmol, 1.0 eq) was added and stirred at 60°C for 3 hours. The reaction solution was concentrated, and the residue was purified by Prep-HPLC (column model: gemini-C18 150x21.2 mm, 5 μm, mobile phase: ACN-H2O (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%).

[0311] LC-MS (ESI), m / z: [M+H] + =429.0.

[0312] 1 H-NMR(400MHz,CD3OD)δ8.66(s,1H),8.39(s,1H),8.05(s,1H),7.66(s,1H),7.00(s,1H ), 4.47 (d, J = 9.2Hz, 2H), 4.20 (d, J = 9.2Hz, 2H), 3.91 (s, 3H), 3.51 (s, 2H), 3.04 (s, 3H).

[0313] The following examples were prepared with reference to the experimental route and method in Example 69:

[0314] Example 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

[0315] Synthesis route:

[0316] Step A: 4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-3-amine

[0317] 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(diphenylphosphino)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 mixture was evaporated to dryness under reduced pressure. The reaction solution was concentrated and the residue was purified by column chromatography (DCM:MeOH=20:1) to give the product 4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-3-amine (131 mg, yield = 58%) as a yellow oil.

[0318] LC-MS (ESI), m / z: [M+H] + =235.0.

[0319] Step B: 2-(3-(3-amino-4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-1-yl)-1-(ethylsulfonyl)azetidin-3-yl)acetonitrile

[0320] 4-(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)acetonitrile (104 mg, 0.59 mmol, 1.0 eq) were dissolved in acetonitrile (5 ml), and 1,8-diazabicyclo[5.4.0]undec-7-ene (85 mg, 0.59 mmol, 1.0 eq) was added dropwise to the reaction solution and stirred at room temperature for 4 h. After completion of the reaction, ice water (20 ml) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL * 3). The combined organic phases were 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 (chromatographic column model: gemini-C18 150 x 19 mm, 5 um, mobile phase: ACN-H2O (0.1% TFA), gradient 10%-55%, flow rate: 20 mL / min) to give the product (200 mg, yield = 80.1%).

[0321] LC-MS (ESI), m / z: [M+H] + =420.8.

[0322] 1 H-NMR (400MHz, CD3OD) δ8.68(s,1H),8.05(s,1H),7.74(s,1H),7.43(s,1H),4.54(d,J=8 .8Hz, 2H), 4.16 (d, J = 9.0Hz, 2H), 3.48 (s, 2H), 3.17-3.11 (m, 2H), 1.33 (t, J = 7.3Hz, 3H).

[0323] 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

[0324] To a solution of 2-(3-(3-amino-4-(6-chloroimidazo[1,2-b]pyridazin-8-yl)-1H-pyrazol-1-yl)-1-(ethylsulfonyl)azacycl-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-butoxide (61 mg, 0.63 mmol, 3.0 eq) in toluene / methanol (1:1, 5 mL) was added methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (5 mg, 0.005 mmol, 0.03 eq). The reaction mixture was stirred at 110°C under nitrogen for 16 h. The reaction mixture was diluted with water (20 mL) and then extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The crude product was purified by Combiflash column (petroleum ether:ethyl acetate = 5:1) to obtain the product (12.8 mg, yield = 13%).

[0325] LC-MS (ESI), m / z: [M+H] + =467.0.

[0326] 1H-NMR (400MHz, CD3OD) δ8.67(s,1H),8.31(s,1H),8.14(s,1H),8.07(d,J=1.2Hz,1H),7.72(d,J=1.2Hz,1H),7.67(s, 1H), 4.61 (d, J = 9.2Hz, 2H), 4.22 (d, J = 9.3Hz, 2H), 4.01 (s, 3H), 3.54 (s, 2H), 3.18 (dt, J = 12.3, 6.2Hz, 2H), 1.38 (m, 3H).

[0327] The following examples were prepared with reference to the experimental route and method in Example 80:

[0328] Example 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

[0329] Synthesis route:

[0330] Step A: Benzyl (1-hydroxypropan-2-yl) carbamate

[0331] 10.0 g (133 mmol, 1.0 eq) of 2-aminopropan-1-ol 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 then slowly added dropwise under an ice bath. The mixture was stirred at room temperature overnight. TLC confirmed the complete reaction of the starting material. 200 mL of water was added, followed by extraction 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 = 10 / 1 to 1:1) to yield the product (13.6 g, yield = 43%).

[0332] LC-MS (ESI), m / z: [M+H] + =210.0.

[0333] 1H-NMR (400MHz, DMSO-d6) δ7.40-7.03(m,5H),7.02(d,J=8.0Hz,1H),5.01(s,2H),4.64(t,J= 5.8Hz,1H),3.48-3.55(m,1H),3.32-3.37(m,1H),3.17-3.23(m,1H),1.02(d,J=6.8Hz,3H).

[0334] Step B: 2-((Benzyloxy)carbonyl)amino)propylmethanesulfonate

[0335] 13.6 g (64.12 mmol, 1.0 eq) of benzyl(1-hydroxypropan-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 under an ice bath. The mixture was stirred at room temperature overnight. TLC confirmed the complete reaction of the starting material. 100 mL of water was added, followed by extraction 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 = 0-2:1) to obtain the product (13.4 g, yield = 60.1%).

[0336] LC-MS (ESI), m / z: [M+H] + =310.0.

[0337] 1 H-NMR (400MHz, CDCl3) δ7.41-7.32(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=7.0Hz,3H).

[0338] Step C: S-(2-((Benzyloxy)carbonyl)amino)propyl)ethylsulfate

[0339] 10.0 g (34.8 mmol, 1.0 eq) of propyl 2-(benzyloxycarbonyl)amino)methanesulfonate 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 thioacetic acid, and the mixture was stirred at room temperature overnight. LC-MS confirmed the formation of the desired product, and 150 mL of water was added. The mixture was then extracted three times with ethyl acetate. The organic layer was washed with 100 mL of saturated saline solution, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The residue was purified by column chromatography (PE / EA = 0-5:1) to afford the product (3.34 g, yield = 28%).

[0340] LC-MS (ESI), m / z: [M+H] + =268.0.

[0341] 1 H-NMR (400MHz, CDCl3) δ8.04 (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 = 6.8Hz, 3H).

[0342] Step D: Benzyl (1-(chlorosulfonyl)propan-2-yl)carbamate

[0343] To a solution of N-chlorosuccinimide (6.68 g, 50 mmol, 3.0 eq) in acetonitrile (20 mL) at 0°C was added a 2M hydrochloric acid solution (3.34 mL, 6.7 mmol, 0.5 eq). The mixture was stirred for 15 minutes. S-(2-((Benzyloxy)carbonyl)amino)propyl)ethylsulfate (3.34 g, 13 mmol, 1.0 eq) was then dissolved in acetonitrile (20 mL) and added dropwise to the mixture. Stir at room temperature for 1 hour. TLC confirmed complete reaction of the starting material. The reaction mixture was diluted with sodium bicarbonate (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to yield 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.

[0344] LC-MS (ESI), m / z: [M+H] + =291.9.

[0345] 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

[0346] To a solution of 2-(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). The reaction mixture was stirred at room temperature for 2 h. LC-MS monitored the reaction. The reaction mixture was spin-dried using an oil pump and the crude product was purified by reverse phase Combiflash column (acetonitrile:water = 3:7) to afford benzyl(1-(chlorosulfonyl)propan-2-yl)carbamate (500 mg, yield = 45%) as a pale yellow solid.

[0347] LC-MS (ESI), m / z: [M+H] + =549.9.

[0348] 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

[0349] 20 mg (0.036 mmol, 1.0 eq) of 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 solution, and 19 mg (0.18 mmol, 5.0 eq) of 10% palladium on carbon was added, and the mixture was stirred at room temperature for 3 h under a hydrogen atmosphere. The formation of the target product was confirmed by LC-MS, and the reaction solution was spin-dried and purified (mobile phase A: 0.1% formic acid + 0.05% triethylamine; mobile phase B: acetonitrile; gradient: 30%-70% B, 55 minutes; flow rate: 70 mL / min), followed by desalting and purification (95% acetonitrile:5% water, 5 minutes, then 90% acetonitrile:70% water:methanol flow rate: 70 mL / min) to give a white solid (3.7 mg, yield = 24%).

[0350] LC-MS: (M+H) + ; m / z = 416.0.

[0351] 1H-NMR (400MHz, CD3OD) δ8.70 (s, 1H), 8.52 (s, 1H), 7.46 (d, J = 3.6Hz, 1H), 6.98 (d, J=4.0Hz,1H),4.68-4.64(m,2H),4.28(d,J=9.2Hz,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 = 6.8Hz, 3H).

[0352] The following examples were prepared with reference to the experimental route and method in Example 86:

[0353] Biological activity experiments

[0354] 1. Enzymatic activity of compounds (IC 50 ) detection

[0355] (1) ROCK1 / 2 detection experimental method

[0356] (a) Use Echo 655 to transfer 50 nL of diluted compound working solution to each well of a reaction plate (784075, Greiner).

[0357] (b) Seal the reaction plate with sealing film and centrifuge at 1000 g for 1 minute.

[0358] (c) Prepare kinase solution.

[0359] (d) Add 5 μL of kinase solution to each well of the reaction plate. Seal the plate with film and centrifuge at 1000 g for 30 seconds. Incubate at room temperature for 10 minutes.

[0360] (e) Prepare a mixture of STK2-substrate-biotin kinase substrate and ATP.

[0361] (f) Add 5 μL of STK2-substrate-biotin and ATP mixture to the reaction plate and centrifuge at 1000 g for 30 seconds to start the reaction.

[0362] (g) ROCK1 kinase was reacted at room temperature for 20 minutes, and ROCK2 kinase was reacted at room temperature for 30 minutes.

[0363] (h) A mixture of Sa-XL 665 (125 nM) and STK-antibody-Cryptate was prepared in HTRF detection buffer.

[0364] (i) Add 10 μL of Sa-XL 665 and STK-antibody-Cryptate mixture to each well, centrifuge at 1000 g for 30 seconds, and incubate at room temperature for 1 hour.

[0365] The signals at 615 nm (Cryptate) and 665 nm (XL665) were read using Envision 2104, and the signal intensity was used to characterize the activity of the kinase.

[0366] The kinase activity data were expressed as the ratio of the kinase activity of the test compound to the kinase activity of the blank group (containing only DMSO), and the IC was obtained by curve fitting using Prism software (GraphPad 7.0). 50 value.

[0367] The specific test results are shown in Table 1 below:

[0368] Table 1 Enzymatic activities of compounds (IC 50 )

[0369] As can be seen from the data in the table, the enzymatic activity of the compounds in the examples of the present invention is better than that of the marketed ROCK inhibitors Ripasudil, Netarsudil and Belumosudil. Some compounds have strong activity against ROCK1 / 2 kinases and can be used to regulate the treatment of ROCK kinase-mediated diseases.

[0370] Among them, Ripasudil has the following structural formula:

[0371] Netarsudil has the following structural formula:

[0372] Belumosudil has the following structural formula:

[0373] 2. Compound cell activity (IC 50 ) Detection (In-Cell Western blotting to detect myosin light chain phosphorylation)

[0374] ROCK phosphorylates myosin light chain at amino acid sites T18 / S19, leading to cytoskeletal changes. Rat smooth muscle cell line A7r5 cells were cultured in DMEM supplemented with 10% FBS. Myosin light chain phosphorylation levels were assessed by in situ immunoblotting using a phopho-MLC-T18 / S19-specific antibody and a secondary detection antibody. Compound-treated cells served as positive controls, while vehicle-treated cells served as negative controls. DRAQ5 was used to stain nuclei as an internal control. Absolute IC50 values ​​were determined using GraphPad Prism 7.0 software using nonlinear regression curve fitting with a variable slope.

[0375] On the first day, A7r5 cells were resuspended in serum-free medium and seeded into PDL-coated 384-well black plates with a clear bottom at a density of 5000 cells per well. Serum starvation culture was performed for 4 hours, and the cells were incubated with the compound in serum-free medium for 1 hour. 50 μL of 8% PFA (paraformaldehyde) was added to each well and fixed at room temperature for 1 hour. The liquid in the well was discarded, and 90 μL of ice methanol was added to each well to permeabilize the cells at 4°C for 1 hour. The plate was then washed 3 times with PBST (0.1% Tween20-PBS) using an automatic dispenser. After patting the plate dry, 50 μL of blocking solution was added to each well and blocked at room temperature for 1 hour. The phspho-MLC-T18 / S19 specific antibody was diluted 1:200 with blocking solution, 20 μL was added to each well, and the film was sealed and incubated at 4°C overnight.

[0376] The next day, the liquid in the wells was discarded and the plates were washed five times with PBST using an automated dispenser. After patting the plates dry, the secondary detection antibody was diluted 1:800 in blocking buffer, 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 then discarded and the plates were washed three times with PBST and three times with ddH2O using an automated dispenser. After patting the plates dry, the plates were scanned using a LICOR Odyssey near-infrared imaging scanner.

[0377] The specific test results are shown in Table 2 below:

[0378] Table 2 Cellular activity of compounds (IC 50 )

[0379] Compared to the marketed ROCK inhibitors Ripasudil, Netarsudil, and Belumosudil, the compounds described in the examples of the present invention exhibit superior cellular activity and are more potent in altering the cytoskeleton by phosphorylating the myosin light chain at amino acid sites T18 / S19. Therefore, the compounds of the present invention have a wider range of applications.

[0380] 3. Effects of the compounds in the SD rat glaucoma model induced by episcleral vein cauterization

[0381] Experimental Methods: After passing quarantine, male Sprague-Dawley rats were obtained and general anesthetized with isoflurane. Topical anesthesia with proparacaine hydrochloride eye drops was administered to the experimental eye (right eye). The eyelids were held open, and the bulbar conjunctiva was incised along the superior limbus. The fascia was bluntly dissected to expose the superior scleral veins. Two superior scleral veins adjacent to the superior rectus muscle and one temporal superior scleral vein were then cauterized. Successful cauterization was indicated by vascular distension proximal to the limbus and absence of blood flow distal to the limbus, with no bleeding. Postoperatively, the conjunctiva was sutured with interrupted 9-0 non-absorbable surgical sutures. Levofloxacin eye drops were administered after suturing, and the rats were returned to their cages after recovery.

[0382] The wound was disinfected with iodine tincture before and after surgery. Levofloxacin eye drops were applied twice a day for one week after surgery. Carprofen was given once 30 minutes before surgery and twice after surgery, with an interval of 24 hours (subcutaneous injection, 10 mg / kg, 1 mL / kg).

[0383] On the third day after modeling, animals were randomly divided into a normal control group, a model control group, a positive control group, and a test substance group, with 5 animals in each group. After modeling, the positive control (commercially available Rhopressa eye drops: 0.02% netarsudil ophthalmic solution) or the test substance (0.01% Example 2) was administered via conjunctival sac eye drops, one drop per day, for 14 consecutive days (a total of 14 doses). The day of the first administration was defined as experimental day 1 (D1).

[0384] During the experiment, the general condition of the animals in each group was observed every day; after administration, the body weight was measured at least once a week, and the intraocular pressure within 24 hours was measured on the 1st, 3rd, 7th, 10th, and 14th days of administration; at the end of administration, the animals were dissected according to the plan for ophthalmological examination, and after euthanasia, gross anatomical observation and histopathological examination were performed (BRN3A immunofluorescence staining: counting the number of RGCs in the retina under a 20x objective lens in one field of view 2 mm away from the optic nerve, and counting the number of RGCs in one circle of the retina in one section).

[0385] Test results:

[0386] Body weight: By the end of the experiment, the body weight of rats in each group showed a steady upward trend, and no significant differences were found in body weight or weight gain among the groups (P>0.05), as shown in Figure 1.

[0387] Intraocular pressure: Data on intraocular pressure changes at different time points (24 hours) on days 1, 3, 7, 10, and 14 of drug administration showed that intraocular pressure in animals in each drug administration group showed a decreasing trend up to 8 hours after drug administration, and was significantly lower at 8 hours compared with the model control group. By 24 hours, intraocular pressure tended to stabilize, while intraocular pressure in animals in the model group remained higher than in all drug administration groups. The intraocular pressure data are shown in Figure 2.

[0388] Histopathology: Compared with the model control group, the number of RGCs in the positive control group and the test substance group increased to a certain extent, as shown in Figure 3.

[0389] The experimental results showed that the compound of the present invention had a certain degree of improvement on glaucoma in the SD rat glaucoma model induced by episcleral vein cauterization, and had a certain protective effect on rat RGCs after eye drops were administered unilaterally into the conjunctival sac of the model eye once a day for 14 consecutive days.

[0390] The present application has been described above in conjunction with preferred embodiments, but these embodiments are merely exemplary and serve only as an illustrative example. On this basis, various replacements 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, polymorph, isotope-labeled substance, isomer or prodrug thereof: in, R 1 NHR 11 、-OR 11 、-SR 11 or -C(=O)NHR 11 ; Among them, R 11 are each independently selected from 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 Heterocyclic, 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 heteroaryl acyl, or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl, thiol, substituted C 3～8 Cycloalkyl, substituted C 3～8 Heterocyclic, substituted C 6～20 Aryl or substituted C 5～20 heteroaryl; R 2 is selected from 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, carboxyl or mercapto; the substituent is selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 3 Selected from cyano, -CONH 2 or carboxyl; L is selected from -S(=O) 2 -, -C(=O)-, -CH 2 -, or -S(=O)(=N)R L -, where R L is selected from hydrogen or substituted or unsubstituted C 1～8 Alkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 4 is selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; The substituent is selected from halogen, hydrazide, C 1～8 Alkylsulfonyl, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, -COR 33 , carboxyl or thiol; 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 One, two or three of are N atoms; the dotted circle represents the bond forming the aromatic ring; R 21 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 22 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 23 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 24 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 25 Selected from hydrogen, halogen, -NR 31 R 32 , substituted or unsubstituted C 3～8 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; Or, R 24 , R 24 Connected W 4 With R 25 and R 25 Connected W 5 , combined together to form A 1 , W 1 , W 2 , W 3 , W 4 and W 5 A 6-membered ring formed by fusion of a 5-membered aryl, cycloalkyl, heteroaryl or heterocycloalkyl ring; R 31 and R 32 are each independently selected from hydrogen, sulfonamide, 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, -C(=O)OR 34 or sulfhydryl; R 33 is selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 ring Alkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, carboxyl or thiol; n is 1, 2 or 3; Furthermore, the compound is not the following compound:

2. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 1 Selected from -NHR 11 , where R 11 are each independently selected from 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 Heterocyclic, 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 heteroaryl acyl, or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl, thiol, substituted C 3～8 Cycloalkyl, substituted C 3～8 Heterocyclic, substituted C 6～20 Aryl or substituted C 5～20 heteroaryl; Among them, R 31 and R 32 are each independently selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, -C(=O)OR 34 or sulfhydryl; R 33 is selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, carboxyl or mercapto.

3. The azacycloalkane compound according to claim 2 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 11 is selected from hydrogen, or substituted or unsubstituted C 1～6 Alkyl, substituted by -NR 31 R 32 、-C(=O)R 33 or substituted or unsubstituted C 3～8 Heterocyclic group.

4. The azacycloalkane compound according to claim 2 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 11 Selected from substituted or unsubstituted C 1～6 Alkyl, substituted by -NR 31 R 32 , where R 31 and R 32 are each independently hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 3～8 Heterocyclic group.

5. The azacycloalkane compound according to claim 2 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 11 Selected from substituted or unsubstituted C 1～6 Alkyl, substituted by -C(=O)R 33 , where R 33 are independently substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 3～8 Heterocyclic group.

6. The azacycloalkane compound according to claim 2 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 1 Selected from -NH 2 、-NHC(=O)CH 3 , or 7. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 1 Select from -OR 11 , where R 11 are each independently selected from 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 Heterocyclic, 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 Heteroaryl acyl or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl or thiol; Preferably, R 11 is selected from hydrogen or substituted or unsubstituted C 1～6 Alkyl, substituted with -NR 31 R 32 、-C(=O)R 33 or substituted or unsubstituted C 3～8 Heterocyclic group; Among them, R 31 and R 32 are each independently selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, -C(=O)OR 34 or sulfhydryl; R 33 is selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, carboxyl or mercapto.

8. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 1 Selected from -C(=O)NHR 11 , where R 11 are each independently selected from 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 Heterocyclic, 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 heteroaryl acyl, or substituted or unsubstituted C 1～6 Alkoxy; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 、-C(=O)R 33 , hydroxyl or thiol; Preferably, R 11 is selected from hydrogen or substituted or unsubstituted C 1～6 Alkyl, substituted by -NR 31 R 32 、-C(=O)R 33 or substituted or unsubstituted C 3～8 Heterocyclic group; Among them, R 31 and R 32 are each independently selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, -C(=O)OR 34 or sulfhydryl; R 33 is selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl may be substituted or unsubstituted Substituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, carboxyl or mercapto.

9. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 2 Selected from hydrogen, R 3 Selected from cyano.

10. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, Selected from the following A1-A7 structures: or Among them, in the A1-A7 structure, R 7 are each independently selected from hydrogen, halogen, -NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 6～20 Aryl, substituted or unsubstituted C 5～20 Heteroaryl or substituted or unsubstituted C 3～8 Heterocyclic group; Wherein the substituent is selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, carboxyl or mercapto.

11. The azacycloalkane compound according to claim 10 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 7 are each independently selected from hydrogen, -NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl; Wherein the substituent is selected from C 1～8 alkyl.

12. The azacycloalkane compound according to claim 10 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, For the following A1 structure: Among them, in the A1 structure, R 7 For hydrogen.

13. The azacycloalkane compound according to claim 10 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, For the following A2 structure: Among them, in the A2 structure, R 7 For-NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 5～20 Heteroaryl or substituted or unsubstituted C 3～8 Heterocyclic group; Wherein the substituent is selected from C 1～8 Alkyl or hydroxyl; or, For the following A3 structure: Among them, in the A3 structure, R 7 For-NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 5～20 Heteroaryl or substituted or unsubstituted C 3～8 Heterocyclic group; Wherein the substituent is selected from C 1～8 Alkyl or hydroxyl; or, For the following A4 structure: Among them, in the A4 structure, R 7 For-NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl, substituted or unsubstituted C 5～20 Heteroaryl or substituted or unsubstituted C 3～8 Heterocyclic group; Wherein the substituent is selected from C 1～8 Alkyl or hydroxyl.

14. The azacycloalkane compound according to claim 10 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, For the following A5 structure: Among them, in the A5 structure, R 7 For-NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl; Wherein the substituent is selected from C 1～8 Alkyl or hydroxyl; or, For the following A6 structure: Among them, in the A6 structure, R 7 Halogen, -NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl; Wherein the substituent is selected from C 1～8 Alkyl or hydroxyl; or, For the following A7 structure: Among them, in the A7 structure, R 7 For-NR 31 R 32 or substituted or unsubstituted C 5～20 Heteroaryl, Among them, R 31 and R 32 are each independently selected from hydrogen, substituted or unsubstituted C 1～6 Alkyl or substituted or unsubstituted C 5～20 heteroaryl; Wherein the substituent is selected from C 1～8 Alkyl or hydroxyl.

15. The azacycloalkane compound according to any one of claims 10-11 or 13-14, or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 7 -NHCH 3 , -NHCH 2 CH 2 OH, Cl, or 16. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, L is selected from -S(=O) 2 -, -C(=O)- or -S(=O)(=N)R L -, where R L Selected from hydrogen or C 1～6 alkyl.

17. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, R 4 Selected from 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 Heteroaryl; substituents selected from halogen, hydrazide, C 1～8 Alkylsulfonyl, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, -COR 33 , carboxyl or thiol; Among them, R 31 and R 32 are each independently selected from hydrogen, sulfonamide, 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 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, cyano, hydroxyl, -C(=O)OR 34 or sulfhydryl; R 33 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Alkoxy, substituted or unsubstituted Generation C 3～8 Cycloalkyl, substituted or unsubstituted C 3～8 Heterocyclic, substituted or unsubstituted C 6～20 Aryl or substituted or unsubstituted C 5～20 heteroaryl; substituents selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 Heteroaryl, cyano, -NR 31 R 32 , hydroxyl, carboxyl or thiol; R 34 is selected from hydrogen, substituted or unsubstituted C 1～8 Alkyl, substituted or unsubstituted C 1～8 Haloalkyl, substituted or unsubstituted C 1～8 Alkoxy or substituted or unsubstituted C 3～8 Cycloalkyl; substituents are selected from halogen, C 1～8 Alkyl, C 1～8 Haloalkyl, C 1～8 Alkoxy, C 3～8 Cycloalkyl, C 3～8 Heterocyclic group, C 6～20 Aryl, C 5～20 heteroaryl, cyano, hydroxyl, carboxyl or mercapto.

18. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, The azacycloalkane compound is selected from the following compounds:

19. The azacycloalkane compound according to claim 1 or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, in, The azacycloalkane compound is selected from the following compounds:

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, polymorph, isotope-labeled substance, 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, polymorph, isotope-labeled substance, isomer or prodrug thereof, and the pharmaceutical composition according to claim 20 in the preparation of a medicament for treating a Rho kinase-mediated disease.

22. The use according to claim 21, in, The Rho kinase-mediated disease is asthma, cancer, glaucoma, insulin resistance, renal failure, neuronal degeneration or osteoporosis.

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

24. The method according to claim 23, in, The Rho kinase-mediated disease is asthma, cancer, glaucoma, insulin resistance, renal failure, neuronal degeneration or osteoporosis.

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