Indole compound, preparation method therefor and use thereof

Abstract

A compound represented by formula I, or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotopic derivative, and pharmaceutically acceptable salt thereof. The compound has activity in preventing and / or treating skin-related diseases.

Description

Indole compound, preparation method and use thereof

[0001] The present disclosure claims priority to the Chinese patent application filed on July 5, 2023, with the invention title “A kind of indole compound, preparation method and use thereof” and application number 202310814278.1, the Chinese patent application filed on March 6, 2024, with the invention title “A kind of indole compound, preparation method and use thereof” and application number 202410257182.4, and the Chinese patent application filed on March 25, 2024, with the invention title “A kind of indole compound, preparation method and use thereof” and application number 202410340315.4. The entire contents of the above applications are incorporated into the present disclosure by reference. Technical Field

[0002] The present invention relates to the field of biomedicine, and more particularly to the use of an indole compound for preventing and / or treating skin-related diseases, especially diseases related to hyperkeratosis of the skin. Background Art

[0003] The epidermis is the outermost layer of the skin, and the stratum corneum is the outermost layer of the epidermis, which evolves from the basal cells of the skin. The stratum corneum is the first line of defense between the human body and the external environment. Its main functions are to retain moisture, obtain mechanical strength, selectively transport molecules, and resist external infections. Hyperkeratosis refers to excessive thickening of the stratum corneum of the mucous membrane or skin, which is mainly related to variations in the production of keratin in the outer layer of the skin. Generally speaking, the increase in the thickness of the stratum corneum is caused by exogenous factors (such as chronic physical or chemical damage) and endogenous factors (such as chronic inflammation and side effects of drugs) (Maria Vastarella et al. Drug Safety 2020, 43(5), 395-408). Hyperkeratosis is common in a variety of skin diseases, such as acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, and various dermatitis. It can also occur as a side effect of various tumor treatments and medications, such as chemotherapy drugs, kinase inhibitors, immunomodulators, and immune checkpoint inhibitors (Austin Greco et al. Cancers 2019, 11(12), 1950; Samantha R. Ellis et al. Journal of the American Academy of Dermatology 2022, 83(4), 1130-1143). Currently, there are no drugs targeting hyperkeratosis, and clinically it can only be alleviated.

[0004] m6A methylation is the most common mRNA modification in mammals, regulating multiple signaling pathways and cellular processes (such as growth, development, and disease), thereby playing a key biological role (Frye M., et al. Science 2018, 361, 2073-2092; Yang C., et al. Cell Death & Disease 2020, 11, 960; Meyer KD & Jaffrey SR Nature Review Molecular Cell Biology 2014, 15, 313-326). m6A methylation of mRNA, miRNA, circRNA, and lncRNA is a dynamic and reversible process. Methyltransferases (such as METTL3, METTL14, and METTL16) attach methyl groups to RNA, while demethylases (FTO and ALKBH5) remove methyl groups from RNA, forming the regulatory basis of m6A. m6A affects RNA processing, translation, and degradation by recruiting specific binding proteins (such as YTHDF1, YTHDF2, YTHDC1, and IGF2BP), thereby leading to changes in downstream protein function and cell biological behavior (Hsu PJ, et al. Journal of Biological Chemistry 2019, 294, 19889-19895; Yao Y., et al. FASEB Journal 2019, 33, 7529-7544).

[0005] Existing studies have shown that m6A is closely associated with the development and progression of skin-related diseases. METTL3 and ALKBH5 mRNA expression levels are significantly higher in tumor tissues of patients with acral melanoma than in adjacent tissues, and METTL3 mRNA expression levels are significantly higher in patients with advanced acral melanoma than in those with early-stage disease (Le Zhanghui, Preliminary Study on the Pathogenesis of LncRNA and RNA m6A Methylation in Acral Melanoma, Dissertation, 2019). In melanoma diagnosis, the m6A-specific binding proteins YTHDF1 and HNRNPA2B1 can serve as novel biomarkers (Li TD, et al. Cancer Cell 2020, 20, 239). In keratinocytes, long-term, low-level arsenic exposure can inhibit m6A expression and induce selective autophagy, thereby inducing skin tumorigenesis (Cui YH, et al. Nature Communications 2021, 12, 2183). After knockout of m6A methyltransferase, the induced proliferation of epidermal cells was inhibited, while the m6A binding protein YTHDC1 can affect epidermal autophagy by regulating the stability of mRNA in keratinocytes (Lee J. et al., EMBO J. 2021, 40(8): e106276; Liang DF et al., Autophage 2022, 18(6): 1318-1337). In a psoriasis mouse model, m6A can regulate the expression of cytokines IL-17A and TNFalpha (Wang Y. et al., Front. Cell. Dev. Biol. 2020, 8, 591629). The m6A level in the skin tissue of psoriasis mice was significantly reduced, while the protein content of m6A demethylase FTO was significantly increased, which played a protective role against psoriasis (Cui L. et al., J. Investig. Dermatol. 2020, 140(7): S6). In addition, m6A methylases, demethylases, and binding proteins are also considered blood markers for systemic lupus erythematosus (Li L. et al., Mol. Immunol. 2018, 93, 55-63). However, there is currently no method to prevent or treat skin diseases by regulating RNA m6A methylation.

[0006] Summary of the Invention

[0007] Problems to be solved by the invention

[0008] In order to solve the above problems, the present invention provides an indole compound, which can be used to prevent and / or treat skin-related diseases.

[0009] Solutions for solving problems

[0010] The present invention provides a compound represented by Formula I or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts:

[0011] in:

[0012] X is selected from NR a , O and S;

[0013] Y is selected from C and N;

[0014] R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 1 replaced by;

[0015] R 2 Selected from hydrogen, deuterium, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 2 replaced by;

[0016] R 3 Selected from hydrogen, deuterium, hydroxyl, -NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 3 replaced by;

[0017] R 4 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 4 replaced by;

[0018] R a is selected from hydrogen, deuterium, alkyl, and haloalkyl;

[0019] Every R bEach independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 5 replaced by;

[0020] Every R c Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 6 replaced by;

[0021] Every R i 、R j are each independently selected from hydrogen, deuterium, -S(O)2-R d , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 7 replaced by;

[0022] Every R d Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 8 replaced by;

[0023] Every R e Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 9 replaced by;

[0024] Q 1 , Q 2 , Q 3 , Q 4 , Q 5 , Q 6 , Q 7 , Q 8 , Q 9 Each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, mercapto, amino, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 10 replaced by;

[0025] Q 10 Selected from hydrogen, deuterium, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl

[0026] m and n are each independently 0, 1 or 2;

[0027] p and q are each independently 0, 1, 2, 3 or 4.

[0028] Effects of the Invention

[0029] The present invention provides a compound shown in Formula I that can be used to prevent and / or treat diseases related to abnormal methylation of RNA m6A, wherein the diseases related to abnormal methylation of RNA m6A are skin-related diseases. The skin-related diseases are diseases related to hyperkeratosis of the skin, including but not limited to rash, pruritus, folliculitis, paronychia, pigmentation disorders, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcers and dermatitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] FIG1 depicts exemplary results of cellular m6A levels and NAP1L2 m6A levels measured by nanopore sequencing 24 hours after administration of sorafenib, capecitabine, docetaxel, and osimertinib to keratinocytes HaCat.

[0031] FIG2 depicts exemplary results of cellular m6A levels and NAP1L2 m6A levels in mouse hind limb skin tissue in Example 15.

[0032] FIG3 shows exemplary results of NAP1L2 mRNA expression measured by RT-PCR 24 hours after administration of human recombinant METTL3 protein, human recombinant FTO protein, or transfection of METTL3 siRNA and FTO siRNA to keratinocytes HaCat.

[0033] FIG4 shows exemplary results of mRNA and protein expression of matrix metalloproteinases measured by RT-PCR and Western blotting 24 hours after administration of human recombinant NAP1L2 protein, sorafenib, capecitabine, docetaxel, and osimertinib, or transfection of NAP1L2 siRNA to keratinocytes HaCat.

[0034] FIG5 depicts exemplary results of mRNA and protein expression levels of keratinocyte differentiation markers KRT1, KRT10, Loricrin, and Involucrin measured by RT-PCR and Western blotting 24 hours after administration of human recombinant HBEGF protein, HBEGF antibody, or transfection of METTL3 and NAP1L2 siRNA to keratinocytes HaCat.

[0035] FIG6 depicts exemplary results of LC-MS determination of cellular m6A methylation inhibition levels 24 hours after administration of compounds 1, 2, 2b, 5a, 8b, nicotinamide, and UZH2 to THP-1 cells.

[0036] FIG7 shows exemplary results of mRNA expression of keratinocyte differentiation markers KRT1, KRT10, Loricrin, and Involucrin measured by RT-PCR 24 hours after administration of human recombinant HBEGF protein and compounds 2, 2b, 3a, 3b, 5a, and 8b to keratinocytes HaCaT.

[0037] FIG8 : depicts exemplary results of the measurement of stratum corneum thickness of rat paw plantar skin in Example 22.

[0038] FIG9 depicts exemplary histopathological results of epithelial blisters, inflammatory cell infiltration, and skin tissue hyperemia in rat paw plantar skin after tissue staining in Example 22.

[0039] FIG10 shows exemplary results of histopathological scoring of epithelial blisters, inflammatory cell infiltration, and skin tissue hyperemia in rat paw plantar skin after tissue staining in Example 22.

[0040] FIG. 11 depicts exemplary results of mouse hind limb skin in Example 23.

[0041] FIG. 12 depicts exemplary results of the degree of swelling in the hind limb toes of mice in Example 23.

[0042] FIG13 shows exemplary histopathological results of mouse hind limb skin tissue in Example 23. FIG.

[0043] FIG14 shows exemplary results of immunohistochemistry for IL-8 in the skin tissues of the hind limbs and toes of mice in the capecitabine model series in Example 23. ...

[0044] FIG15 shows exemplary results of immunohistochemistry for IL-6 in the skin tissues of the hind limbs and toes of mice in the docetaxel modeling series in Example 23.

[0045] FIG. 16 depicts exemplary results of mouse hind limb skin in Example 24.

[0046] FIG. 17 depicts exemplary results of the degree of swelling in the hind limb toes of mice in Example 24.

[0047] FIG. 18 shows exemplary histopathological results of mouse hind limb skin tissue in Example 24.

[0048] FIG19 shows exemplary results of immunohistochemistry for IL-1β in the skin tissues of the hind limbs and toes of mice in the Sorafenib model series in Example 24.

[0049] Figure 20 shows exemplary results of immunohistochemistry for IL-1β in the skin tissues of the hind limbs and toes of mice in the osimertinib model series in Example 24. DETAILED DESCRIPTION

[0050] To make the technical solutions and beneficial effects of the present invention more clearly understood, the following detailed description is given by way of specific embodiments. The accompanying drawings are not necessarily drawn to scale, and local features may be enlarged or reduced to more clearly illustrate the details of the local features. Unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application belongs.

[0051] In one aspect, the present invention provides a compound of Formula I or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives, and pharmaceutically acceptable salts:

[0052] in:

[0053] X is selected from NR a , O and S;

[0054] Y is selected from C and N;

[0055] R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 1 replaced by;

[0056] R 2 Selected from hydrogen, deuterium, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 2 replaced by;

[0057] R 3 Selected from hydrogen, deuterium, hydroxyl, -NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 3 replaced by;

[0058] R 4Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 4 replaced by;

[0059] R a is selected from hydrogen, deuterium, alkyl, and haloalkyl;

[0060] Every R b Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 5 replaced by;

[0061] Every R c Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 6 replaced by;

[0062] Every R i 、R j are each independently selected from hydrogen, deuterium, -S(O)2-R d , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 7 replaced by;

[0063] Every R d Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 8 replaced by;

[0064] Every R e Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 9 replaced by;

[0065] Q 1 , Q 2 , Q 3 , Q 4 , Q 5 , Q 6 , Q 7 , Q 8 , Q 9Each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, mercapto, amino, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 10 replaced by;

[0066] Q 10 is selected from the group consisting of hydrogen, deuterium, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl;

[0067] m and n are each independently 0, 1 or 2;

[0068] p and q are each independently 0, 1, 2, 3 or 4.

[0069] In certain embodiments, X is selected from NR a and O.

[0070] In certain embodiments, X is selected from NR a .

[0071] In certain embodiments, the R a is selected from hydrogen, deuterium and alkyl.

[0072] In certain embodiments, the R a Selected from hydrogen and C 1-3 alkyl.

[0073] In certain embodiments, the R a is selected from hydrogen and methyl.

[0074] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups.

[0075] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclyl.

[0076] In certain embodiments, the R 1 Selected from hydrogen, deuterium, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl.

[0077] In certain embodiments, the R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , substituted or unsubstituted methyl, cyclopropyl.

[0078] In certain embodiments, the Q 1 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0079] In certain embodiments, the Q 1 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0080] In certain embodiments, the Q 1 Selected from hydrogen and halogen.

[0081] In certain embodiments, the Q 1 Selected from hydrogen.

[0082] In certain embodiments, the R b is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0083] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl.

[0084] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0085] In certain embodiments, the R b is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0086] In certain embodiments, the Q 5 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0087] In certain embodiments, the Q 5 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0088] In certain embodiments, the Q 5is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0089] In certain embodiments, the Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0090] In certain embodiments, the R c is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0091] In certain embodiments, the R c Selected from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl.

[0092] In certain embodiments, the R c Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0093] In certain embodiments, the R c is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0094] In certain embodiments, the Q 6 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0095] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0096] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0097] In certain embodiments, the Q 6 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0098] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0099] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10aryl and 5-10 membered heteroaryl.

[0100] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0101] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0102] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0103] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0104] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0105] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0106] In certain embodiments, the Q 7 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0107] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0108] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0109] In certain embodiments, the Q 7 is selected from hydrogen and pyridyl.

[0110] In certain embodiments, the R d is selected from hydrogen, substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0111] In certain embodiments, the R d is selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0112] In certain embodiments, the R d Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl and C 6-10 Aryl.

[0113] In certain embodiments, the R d Selected from methyl, cyclopropyl and phenyl.

[0114] In certain embodiments, the Q 8 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0115] In certain embodiments, the Q 8 is selected from hydrogen, halogen and amino.

[0116] In certain embodiments, the Q 8 Selected from hydrogen, fluorine and chlorine.

[0117] In certain embodiments, the Q 8 Selected from hydrogen.

[0118] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0119] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-Re , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl.

[0120] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl.

[0121] In certain embodiments, the R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl.

[0122] In certain embodiments, the Q 2 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0123] In certain embodiments, the Q 2 is selected from hydrogen, halogen and amino.

[0124] In certain embodiments, the Q 2 Selected from hydrogen, fluorine and chlorine.

[0125] In certain embodiments, the Q 2 Selected from hydrogen.

[0126] In certain embodiments, the R e is selected from hydrogen, substituted or unsubstituted alkyl, aryl and heterocyclic groups.

[0127] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 3-10 membered heterocyclic group.

[0128] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 aryl and 3-6 membered heterocyclic group.

[0129] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl.

[0130] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, aryl and heteroaryl.

[0131] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0132] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S.

[0133] In certain embodiments, the Q 9 Selected from hydrogen, fluorine, methyl, pyrrolyl.

[0134] In certain embodiments, the Q 10 is selected from hydrogen, halogen, and substituted or unsubstituted alkyl.

[0135] In certain embodiments, the Q 10 Selected from hydrogen and halogen.

[0136] In certain embodiments, the Q 10 Selected from hydrogen and fluorine.

[0137] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j , substituted or unsubstituted alkyl and alkoxy.

[0138] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j .

[0139] In certain embodiments, the R 3 Selected from-NR i R j .

[0140] In certain embodiments, the Q 3 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0141] In certain embodiments, the Q 3 is selected from hydrogen, halogen and amino.

[0142] In certain embodiments, the Q 3 Selected from hydrogen, fluorine and chlorine.

[0143] In certain embodiments, the Q 3 Selected from hydrogen.

[0144] In certain embodiments, the R 4 Selected from hydrogen, halogen, -OR b and -NR i R j .

[0145] In certain embodiments, the R 4 Selected from hydrogen and halogen.

[0146] In certain embodiments, the R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine.

[0147] In certain embodiments, the R 4 Selected from hydrogen and fluorine.

[0148] In certain embodiments, the Q 4 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0149] In certain embodiments, the Q 4 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0150] In certain embodiments, the Q 4 Selected from hydrogen and halogen.

[0151] In certain embodiments, the Q 4 Selected from hydrogen.

[0152] In certain embodiments, m is 0 or 1.

[0153] In certain embodiments, n is 1 or 2.

[0154] In certain embodiments, p is 1, 2 or 3.

[0155] In certain embodiments, q is 0, 1 or 2.

[0156] In certain embodiments, X is selected from NR a ;

[0157] The R a Selected from hydrogen and C 1-3 alkyl;

[0158] The R 1 Selected from hydrogen, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10Aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl; said substitution is by one or more Q 1 replaced by;

[0159] The Q 1 is selected from hydrogen and halogen;

[0160] The R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl; said substitution is by one or more Q 5 replaced by;

[0161] The Q 5 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0162] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0163] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0164] The Q 7 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0165] The R d Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-5 Cycloalkyl and C 6-10 Aryl; said substitution is by one or more Q 8 replaced by;

[0166] The Q8 is selected from hydrogen, fluorine and chlorine;

[0167] The R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl;

[0168] The R e Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 Aryl and 3-6 membered heterocyclic group; said substitution is by one or more Q 9 replaced by;

[0169] The Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S; said substitution is by one or more Q 10 replaced by;

[0170] The Q 10 selected from hydrogen and halogen;

[0171] The R 3 Selected from-NR i R j ;

[0172] The R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine;

[0173] The m is 0 or 1;

[0174] Said n is 1 or 2;

[0175] Said p is 1, 2 or 3;

[0176] The q is 0, 1 or 2.

[0177] In certain embodiments, X is selected from NR a ;

[0178] The R a is selected from hydrogen and methyl;

[0179] The R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , methyl, cyclopropyl;

[0180] The R bis selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl; said substitution is by one or more Q 5 replaced by;

[0181] The Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl; said substitution is by one or more Q 10 replaced by;

[0182] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0183] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0184] The Q 7 is selected from the group consisting of hydrogen and pyridyl;

[0185] The R d is selected from methyl, cyclopropyl and phenyl;

[0186] The R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl;

[0187] The R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl; said substitution is by one or more Q 9 replaced by;

[0188] The Q 9 is selected from hydrogen, fluorine, methyl, pyrrolyl;

[0189] The Q 10 selected from hydrogen and fluorine;

[0190] The R 3 Selected from-NR i R j ;

[0191] The R 4 is selected from hydrogen and fluorine;

[0192] The m is 0 or 1;

[0193] Said n is 1 or 2;

[0194] Said p is 1, 2 or 3;

[0195] The q is 0, 1 or 2.

[0196] In another aspect, the present invention provides a compound represented by formula I-1 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives, and pharmaceutically acceptable salts:

[0197] in:

[0198] X is selected from NR a , O and S;

[0199] R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 1 replaced by;

[0200] R 2 Selected from hydrogen, deuterium, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 2 replaced by;

[0201] R 3 Selected from hydrogen, deuterium, hydroxyl, -NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 3 replaced by;

[0202] R 4 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 4 replaced by;

[0203] R a is selected from hydrogen, deuterium, alkyl, and haloalkyl;

[0204] Every R b Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 5 replaced by;

[0205] Every R c Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 6 replaced by;

[0206] Every R i 、R j are each independently selected from hydrogen, deuterium, -S(O)2-R d , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 7 replaced by;

[0207] Every R d Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 8 replaced by;

[0208] Every R e Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 9 replaced by;

[0209] Q 1 , Q 2 , Q 3 , Q 4 , Q 5 , Q 6 , Q 7 , Q 8 , Q 9 Each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, mercapto, amino, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 10 replaced by;

[0210] Q 10 is selected from the group consisting of hydrogen, deuterium, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl;

[0211] p and q are each independently 0, 1, 2, 3 or 4.

[0212] In certain embodiments, X is selected from NR a and O.

[0213] In certain embodiments, X is selected from NR a .

[0214] In certain embodiments, the R a is selected from hydrogen, deuterium and alkyl.

[0215] In certain embodiments, the R a Selected from hydrogen and C 1-3 alkyl.

[0216] In certain embodiments, the R a is selected from hydrogen and methyl.

[0217] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups.

[0218] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclyl.

[0219] In certain embodiments, the R 1 Selected from hydrogen, deuterium, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl.

[0220] In certain embodiments, the R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , substituted or unsubstituted methyl, cyclopropyl.

[0221] In certain embodiments, the Q 1is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0222] In certain embodiments, the Q 1 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0223] In certain embodiments, the Q 1 Selected from hydrogen and halogen.

[0224] In certain embodiments, the Q 1 Selected from hydrogen.

[0225] In certain embodiments, the R b is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0226] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl.

[0227] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0228] In certain embodiments, the R b is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0229] In certain embodiments, the Q 5 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0230] In certain embodiments, the Q 5 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0231] In certain embodiments, the Q 5 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0232] In certain embodiments, the Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0233] In certain embodiments, the R c is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0234] In certain embodiments, the R c Selected from substituted or unsubstituted C1-6 Alkyl and C 3-7 Cycloalkyl.

[0235] In certain embodiments, the R c Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0236] In certain embodiments, the R c is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0237] In certain embodiments, the Q 6 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0238] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0239] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0240] In certain embodiments, the Q 6 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0241] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0242] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0243] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0244] In certain embodiments, the R i Selected from hydrogen, -S(O)2-Rd , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0245] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0246] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0247] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0248] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0249] In certain embodiments, the Q 7 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0250] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0251] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0252] In certain embodiments, the Q 7 is selected from hydrogen and pyridyl.

[0253] In certain embodiments, the R d is selected from hydrogen, substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0254] In certain embodiments, the Rd Selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0255] In certain embodiments, the R d Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl and C 6-10 Aryl.

[0256] In certain embodiments, the R d Selected from methyl, cyclopropyl and phenyl.

[0257] In certain embodiments, the Q 8 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0258] In certain embodiments, the Q 8 is selected from hydrogen, halogen and amino.

[0259] In certain embodiments, the Q 8 Selected from hydrogen, fluorine and chlorine.

[0260] In certain embodiments, the Q 8 Selected from hydrogen.

[0261] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0262] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl.

[0263] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl.

[0264] In certain embodiments, the R 2Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl.

[0265] In certain embodiments, the Q 2 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0266] In certain embodiments, the Q 2 is selected from hydrogen, halogen and amino.

[0267] In certain embodiments, the Q 2 Selected from hydrogen, fluorine and chlorine.

[0268] In certain embodiments, the Q 2 Selected from hydrogen.

[0269] In certain embodiments, the R e is selected from hydrogen, substituted or unsubstituted alkyl, aryl and heterocyclic groups.

[0270] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 3-10 membered heterocyclic group.

[0271] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 aryl and 3-6 membered heterocyclic group.

[0272] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl.

[0273] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, aryl and heteroaryl.

[0274] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0275] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S.

[0276] In certain embodiments, the Q9 Selected from hydrogen, fluorine, methyl, pyrrolyl.

[0277] In certain embodiments, the Q 10 is selected from hydrogen, halogen, and substituted or unsubstituted alkyl.

[0278] In certain embodiments, the Q 10 Selected from hydrogen and halogen.

[0279] In certain embodiments, the Q 10 Selected from hydrogen and fluorine.

[0280] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j , substituted or unsubstituted alkyl and alkoxy.

[0281] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j .

[0282] In certain embodiments, the R 3 Selected from-NR i R j .

[0283] In certain embodiments, the Q 3 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0284] In certain embodiments, the Q 3 is selected from hydrogen, halogen and amino.

[0285] In certain embodiments, the Q 3 Selected from hydrogen, fluorine and chlorine.

[0286] In certain embodiments, the Q 3 Selected from hydrogen.

[0287] In certain embodiments, the R 4 Selected from hydrogen, halogen, -OR b and -NR i R j .

[0288] In certain embodiments, the R 4 Selected from hydrogen and halogen.

[0289] In certain embodiments, the R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine.

[0290] In certain embodiments, the R4 Selected from hydrogen and fluorine.

[0291] In certain embodiments, the Q 4 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0292] In certain embodiments, the Q 4 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0293] In certain embodiments, the Q 4 Selected from hydrogen and halogen.

[0294] In certain embodiments, the Q 4 Selected from hydrogen.

[0295] In certain embodiments, p is 1, 2 or 3.

[0296] In certain embodiments, q is 0, 1 or 2.

[0297] In certain embodiments, X is selected from NR a ;

[0298] The R a Selected from hydrogen and C 1-3 alkyl;

[0299] The R 1 Selected from hydrogen, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl; said substitution is by one or more Q 1 replaced by;

[0300] The Q 1 is selected from hydrogen and halogen;

[0301] The R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl; said substitution is by one or more Q 5 replaced by;

[0302] The Q 5 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0303] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0304] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0305] The Q 7 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0306] The R d Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-5 Cycloalkyl and C 6-10 Aryl; said substitution is by one or more Q 8 replaced by;

[0307] The Q 8 is selected from hydrogen, fluorine and chlorine;

[0308] The R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl;

[0309] The R e Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 Aryl and 3-6 membered heterocyclic group; said substitution is by one or more Q 9 replaced by;

[0310] The Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S; said substitution is by one or more Q 10 replaced by;

[0311] The Q 10 selected from hydrogen and halogen;

[0312] The R 3 Selected from-NR i R j ;

[0313] The R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine;

[0314] Said p is 1, 2 or 3;

[0315] The q is 0, 1 or 2.

[0316] In certain embodiments, X is selected from NR a ;

[0317] The R a is selected from hydrogen and methyl;

[0318] The R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , methyl, cyclopropyl;

[0319] The R b is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl; said substitution is by one or more Q 5 replaced by;

[0320] The Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl; said substitution is by one or more Q 10 replaced by;

[0321] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0322] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0323] The Q 7is selected from the group consisting of hydrogen and pyridyl;

[0324] The R d is selected from methyl, cyclopropyl and phenyl;

[0325] The R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl;

[0326] The R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl; said substitution is by one or more Q 9 replaced by;

[0327] The Q 9 is selected from hydrogen, fluorine, methyl, pyrrolyl;

[0328] The Q 10 selected from hydrogen and fluorine;

[0329] The R 3 Selected from-NR i R j ;

[0330] The R 4 is selected from hydrogen and fluorine;

[0331] Said p is 1, 2 or 3;

[0332] The q is 0, 1 or 2.

[0333] In another aspect, the present invention provides a compound represented by formula I-2 or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof:

[0334] in:

[0335] X is selected from NR a , O and S;

[0336] R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 1 replaced by;

[0337] R 2 Selected from hydrogen, deuterium, halogen, -S(O)2-R d、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 2 replaced by;

[0338] R 3 Selected from hydrogen, deuterium, hydroxyl, -NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 3 replaced by;

[0339] R 4 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 4 replaced by;

[0340] R a is selected from hydrogen, deuterium, alkyl, and haloalkyl;

[0341] Every R b Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 5 replaced by;

[0342] Every R c Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 6 replaced by;

[0343] Every R i 、R j are each independently selected from hydrogen, deuterium, -S(O)2-R d , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 7 replaced by;

[0344] Every R d Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 8 replaced by;

[0345] Every R e Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 9 replaced by;

[0346] Q 1 , Q 2 , Q 3 , Q 4 , Q 5 , Q 6 , Q 7 , Q 8 , Q 9 Each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, mercapto, amino, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 10 replaced by;

[0347] Q 10 is selected from the group consisting of hydrogen, deuterium, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl;

[0348] p and q are each independently 0, 1, 2, 3 or 4.

[0349] In certain embodiments, X is selected from NR a and O.

[0350] In certain embodiments, X is selected from NR a .

[0351] In certain embodiments, the R a is selected from hydrogen, deuterium and alkyl.

[0352] In certain embodiments, the R a Selected from hydrogen and C 1-3 alkyl.

[0353] In certain embodiments, the R a is selected from hydrogen and methyl.

[0354] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups.

[0355] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b、-NR i R j , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclyl.

[0356] In certain embodiments, the R 1 Selected from hydrogen, deuterium, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl.

[0357] In certain embodiments, the R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , substituted or unsubstituted methyl, cyclopropyl.

[0358] In certain embodiments, the Q 1 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0359] In certain embodiments, the Q 1 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0360] In certain embodiments, the Q 1 Selected from hydrogen and halogen.

[0361] In certain embodiments, the Q 1 Selected from hydrogen.

[0362] In certain embodiments, the R b is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0363] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl.

[0364] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0365] In certain embodiments, the R bis selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0366] In certain embodiments, the Q 5 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0367] In certain embodiments, the Q 5 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0368] In certain embodiments, the Q 5 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0369] In certain embodiments, the Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0370] In certain embodiments, the R c is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0371] In certain embodiments, the R c Selected from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl.

[0372] In certain embodiments, the R c Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0373] In certain embodiments, the R c is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0374] In certain embodiments, the Q 6 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0375] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0376] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0377] In certain embodiments, the Q 6 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0378] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0379] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0380] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0381] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0382] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0383] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0384] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0385] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d, substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0386] In certain embodiments, the Q 7 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0387] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0388] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0389] In certain embodiments, the Q 7 is selected from hydrogen and pyridyl.

[0390] In certain embodiments, the R d is selected from hydrogen, substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0391] In certain embodiments, the R d Selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0392] In certain embodiments, the R d Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl and C 6-10 Aryl.

[0393] In certain embodiments, the R d Selected from methyl, cyclopropyl and phenyl.

[0394] In certain embodiments, the Q 8 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0395] In certain embodiments, the Q 8 is selected from hydrogen, halogen and amino.

[0396] In certain embodiments, the Q 8 Selected from hydrogen, fluorine and chlorine.

[0397] In certain embodiments, the Q 8 Selected from hydrogen.

[0398] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0399] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl.

[0400] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl.

[0401] In certain embodiments, the R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl.

[0402] In certain embodiments, the Q 2 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0403] In certain embodiments, the Q 2 is selected from hydrogen, halogen and amino.

[0404] In certain embodiments, the Q 2 Selected from hydrogen, fluorine and chlorine.

[0405] In certain embodiments, the Q 2 Selected from hydrogen.

[0406] In certain embodiments, the R e is selected from hydrogen, substituted or unsubstituted alkyl, aryl and heterocyclic groups.

[0407] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 3-10 membered heterocyclic group.

[0408] In certain embodiments, the R eSelected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 aryl and 3-6 membered heterocyclic group.

[0409] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl.

[0410] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, aryl and heteroaryl.

[0411] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0412] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S.

[0413] In certain embodiments, the Q 9 Selected from hydrogen, fluorine, methyl, pyrrolyl.

[0414] In certain embodiments, the Q 10 is selected from hydrogen, halogen, and substituted or unsubstituted alkyl.

[0415] In certain embodiments, the Q 10 Selected from hydrogen and halogen.

[0416] In certain embodiments, the Q 10 Selected from hydrogen and fluorine.

[0417] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j , substituted or unsubstituted alkyl and alkoxy.

[0418] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j .

[0419] In certain embodiments, the R 3 Selected from-NR i R j .

[0420] In certain embodiments, the Q 3Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0421] In certain embodiments, the Q 3 is selected from hydrogen, halogen and amino.

[0422] In certain embodiments, the Q 3 Selected from hydrogen, fluorine and chlorine.

[0423] In certain embodiments, the Q 3 Selected from hydrogen.

[0424] In certain embodiments, the R 4 Selected from hydrogen, halogen, -OR b and -NR i R j .

[0425] In certain embodiments, the R 4 Selected from hydrogen and halogen.

[0426] In certain embodiments, the R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine.

[0427] In certain embodiments, the R 4 Selected from hydrogen and fluorine.

[0428] In certain embodiments, the Q 4 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0429] In certain embodiments, the Q 4 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0430] In certain embodiments, the Q 4 Selected from hydrogen and halogen.

[0431] In certain embodiments, the Q 4 Selected from hydrogen.

[0432] In certain embodiments, p is 1, 2 or 3.

[0433] In certain embodiments, q is 0, 1 or 2.

[0434] In certain embodiments, X is selected from NR a ;

[0435] The R a Selected from hydrogen and C 1-3 alkyl;

[0436] The R 1Selected from hydrogen, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl; said substitution is by one or more Q 1 replaced by;

[0437] The Q 1 is selected from hydrogen and halogen;

[0438] The R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl; said substitution is by one or more Q 5 replaced by;

[0439] The Q 5 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0440] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0441] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0442] The Q 7 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0443] The R dSelected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-5 Cycloalkyl and C 6-10 Aryl; said substitution is by one or more Q 8 replaced by;

[0444] The Q 8 is selected from hydrogen, fluorine and chlorine;

[0445] The R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl;

[0446] The R e Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 Aryl and 3-6 membered heterocyclic group; said substitution is by one or more Q 9 replaced by;

[0447] The Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S; said substitution is by one or more Q 10 replaced by;

[0448] The Q 10 selected from hydrogen and halogen;

[0449] The R 3 Selected from-NR i R j ;

[0450] The R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine;

[0451] Said p is 1, 2 or 3;

[0452] The q is 0, 1 or 2.

[0453] In certain embodiments, X is selected from NR a ;

[0454] The R a is selected from hydrogen and methyl;

[0455] The R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , methyl, cyclopropyl;

[0456] The R b is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl; said substitution is by one or more Q 5 replaced by;

[0457] The Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl; said substitution is by one or more Q 10 replaced by;

[0458] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0459] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0460] The Q 7 is selected from the group consisting of hydrogen and pyridyl;

[0461] The R d is selected from methyl, cyclopropyl and phenyl;

[0462] The R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl;

[0463] The R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl; said substitution is by one or more Q 9 replaced by;

[0464] The Q 9 is selected from hydrogen, fluorine, methyl, pyrrolyl;

[0465] The Q 10 selected from hydrogen and fluorine;

[0466] The R 3 Selected from-NR i R j ;

[0467] The R 4 is selected from hydrogen and fluorine;

[0468] Said p is 1, 2 or 3;

[0469] The q is 0, 1 or 2.

[0470] In another aspect, the present invention provides a compound represented by formula I-3 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives, and pharmaceutically acceptable salts:

[0471] in:

[0472] X is selected from NR a , O and S;

[0473] R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 1 replaced by;

[0474] R 2 Selected from hydrogen, deuterium, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 2 replaced by;

[0475] R 3 Selected from hydrogen, deuterium, hydroxyl, -NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 3 replaced by;

[0476] R 4 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 4 replaced by;

[0477] R a is selected from hydrogen, deuterium, alkyl, and haloalkyl;

[0478] Every R bEach independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 5 replaced by;

[0479] Every R c Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 6 replaced by;

[0480] Every R i 、R j are each independently selected from hydrogen, deuterium, -S(O)2-R d , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups; the substitution is by one or more Q 7 replaced by;

[0481] Every R d Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 8 replaced by;

[0482] Every R e Each independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 9 replaced by;

[0483] Q 1 , Q 2 , Q 3 , Q 4 , Q 5 , Q 6 , Q 7 , Q 8 , Q 9 Each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, mercapto, amino, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; said substitution is by one or more Q 10 replaced by;

[0484] Q 10 is selected from the group consisting of hydrogen, deuterium, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl;

[0485] p and q are each independently 0, 1, 2, 3 or 4.

[0486] In certain embodiments, X is selected from NRa and O.

[0487] In certain embodiments, X is selected from NR a .

[0488] In certain embodiments, the R a is selected from hydrogen, deuterium and alkyl.

[0489] In certain embodiments, the R a Selected from hydrogen and C 1-3 alkyl.

[0490] In certain embodiments, the R a is selected from hydrogen and methyl.

[0491] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic groups.

[0492] In certain embodiments, the R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclyl.

[0493] In certain embodiments, the R 1 Selected from hydrogen, deuterium, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl.

[0494] In certain embodiments, the R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , substituted or unsubstituted methyl, cyclopropyl.

[0495] In certain embodiments, the Q 1 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0496] In certain embodiments, the Q1 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0497] In certain embodiments, the Q 1 Selected from hydrogen and halogen.

[0498] In certain embodiments, the Q 1 Selected from hydrogen.

[0499] In certain embodiments, the R b is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0500] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl.

[0501] In certain embodiments, the R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0502] In certain embodiments, the R b is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0503] In certain embodiments, the Q 5 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0504] In certain embodiments, the Q 5 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0505] In certain embodiments, the Q 5 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0506] In certain embodiments, the Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0507] In certain embodiments, the R c is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl.

[0508] In certain embodiments, the R c Selected from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl.

[0509] In certain embodiments, the R c Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl.

[0510] In certain embodiments, the R c is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

[0511] In certain embodiments, the Q 6 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0512] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0513] In certain embodiments, the Q 6 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0514] In certain embodiments, the Q 6 is selected from hydrogen and substituted or unsubstituted pyridyl.

[0515] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0516] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0517] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0518] In certain embodiments, the R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0519] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0520] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0521] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0522] In certain embodiments, the R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

[0523] In certain embodiments, the Q 7 is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted heteroaryl.

[0524] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-10 membered heteroaryl.

[0525] In certain embodiments, the Q 7 is selected from hydrogen, halogen, and substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S.

[0526] In certain embodiments, the Q 7 is selected from hydrogen and pyridyl.

[0527] In certain embodiments, the R d is selected from hydrogen, substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0528] In certain embodiments, the R d Selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C6-10 aryl and 5-10 membered heteroaryl.

[0529] In certain embodiments, the R d Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl and C 6-10 Aryl.

[0530] In certain embodiments, the R d Selected from methyl, cyclopropyl and phenyl.

[0531] In certain embodiments, the Q 8 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0532] In certain embodiments, the Q 8 is selected from hydrogen, halogen and amino.

[0533] In certain embodiments, the Q 8 Selected from hydrogen, fluorine and chlorine.

[0534] In certain embodiments, the Q 8 Selected from hydrogen.

[0535] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl.

[0536] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl.

[0537] In certain embodiments, the R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl.

[0538] In certain embodiments, the R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl.

[0539] In certain embodiments, the Q 2 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0540] In certain embodiments, the Q 2 is selected from hydrogen, halogen and amino.

[0541] In certain embodiments, the Q 2 Selected from hydrogen, fluorine and chlorine.

[0542] In certain embodiments, the Q 2 Selected from hydrogen.

[0543] In certain embodiments, the R e is selected from hydrogen, substituted or unsubstituted alkyl, aryl and heterocyclic groups.

[0544] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 3-10 membered heterocyclic group.

[0545] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 aryl and 3-6 membered heterocyclic group.

[0546] In certain embodiments, the R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl.

[0547] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, aryl and heteroaryl.

[0548] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 aryl and 5-10 membered heteroaryl.

[0549] In certain embodiments, the Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S.

[0550] In certain embodiments, the Q 9 Selected from hydrogen, fluorine, methyl, pyrrolyl.

[0551] In certain embodiments, the Q 10is selected from hydrogen, halogen, and substituted or unsubstituted alkyl.

[0552] In certain embodiments, the Q 10 Selected from hydrogen and halogen.

[0553] In certain embodiments, the Q 10 Selected from hydrogen and fluorine.

[0554] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j , substituted or unsubstituted alkyl and alkoxy.

[0555] In certain embodiments, the R 3 Selected from hydrogen, hydroxyl, -NR i R j .

[0556] In certain embodiments, the R 3 Selected from-NR i R j .

[0557] In certain embodiments, the Q 3 Selected from hydrogen, halogen, cyano, hydroxy, mercapto and amino.

[0558] In certain embodiments, the Q 3 is selected from hydrogen, halogen and amino.

[0559] In certain embodiments, the Q 3 Selected from hydrogen, fluorine and chlorine.

[0560] In certain embodiments, the Q 3 Selected from hydrogen.

[0561] In certain embodiments, the R 4 Selected from hydrogen, halogen, -OR b and -NR i R j .

[0562] In certain embodiments, the R 4 Selected from hydrogen and halogen.

[0563] In certain embodiments, the R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine.

[0564] In certain embodiments, the R 4 Selected from hydrogen and fluorine.

[0565] In certain embodiments, the Q 4is selected from hydrogen, halogen, cyano, hydroxy, amino, and substituted or unsubstituted alkyl.

[0566] In certain embodiments, the Q 4 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl.

[0567] In certain embodiments, the Q 4 Selected from hydrogen and halogen.

[0568] In certain embodiments, the Q 4 Selected from hydrogen.

[0569] In certain embodiments, p is 1, 2 or 3.

[0570] In certain embodiments, q is 0, 1 or 2.

[0571] In certain embodiments, X is selected from NR a ;

[0572] The R a Selected from hydrogen and C 1-3 alkyl;

[0573] The R 1 Selected from hydrogen, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl; said substitution is by one or more Q 1 replaced by;

[0574] The Q 1 is selected from hydrogen and halogen;

[0575] The R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl; said substitution is by one or more Q 5 replaced by;

[0576] The Q 5 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0577] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0578] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by;

[0579] The Q 7 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by;

[0580] The R d Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-5 Cycloalkyl and C 6-10 Aryl; said substitution is by one or more Q 8 replaced by;

[0581] The Q 8 is selected from hydrogen, fluorine and chlorine;

[0582] The R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、C 1-3 Alkyl, C 3-6 Cycloalkyl;

[0583] The R e Selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 Aryl and 3-6 membered heterocyclic group; said substitution is by one or more Q 9 replaced by;

[0584] The Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S; said substitution is by one or more Q 10 replaced by;

[0585] The Q 10 selected from hydrogen and halogen;

[0586] The R 3 Selected from-NR i R j ;

[0587] The R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine;

[0588] Said p is 1, 2 or 3;

[0589] The q is 0, 1 or 2.

[0590] In certain embodiments, X is selected from NR a ;

[0591] The R a is selected from hydrogen and methyl;

[0592] The R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , methyl, cyclopropyl;

[0593] The R b is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl; said substitution is by one or more Q 5 replaced by;

[0594] The Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl; said substitution is by one or more Q 10 replaced by;

[0595] The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0596] The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by;

[0597] The Q 7 is selected from the group consisting of hydrogen and pyridyl;

[0598] The R d is selected from methyl, cyclopropyl and phenyl;

[0599] The R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl;

[0600] The R e Selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl; said substitution is by one or more Q 9 replaced by;

[0601] The Q 9 is selected from hydrogen, fluorine, methyl, pyrrolyl;

[0602] The Q 10 selected from hydrogen and fluorine;

[0603] The R 3 Selected from-NR i R j ;

[0604] The R 4 is selected from hydrogen and fluorine;

[0605] Said p is 1, 2 or 3;

[0606] The q is 0, 1 or 2.

[0607] In another aspect, the present invention provides a compound or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof, wherein the compound is any one of the following:

[0608] The present invention also provides a method for preparing the compound represented by formula I, which is as follows:

[0609] After mixing raw materials A and B, heat to 100-200°C and react for 3-8 hours. After cooling to room temperature, extract and concentrate with an appropriate solvent (such as dichloromethane), add LiOH solution, stir at room temperature for 1-3 hours, extract and concentrate again, add a reaction reagent (such as ammonia water), heat to 50-100°C and react for 12-36 hours, and separate and purify to obtain the compound shown in Formula I.

[0610] The present invention also provides a pharmaceutical composition comprising the aforementioned compound or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts.

[0611] In certain embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

[0612] In certain embodiments, the pharmaceutical composition comprises 0.01% to 99.99% of the aforementioned compound, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition comprises 0.1% to 99.9% of the aforementioned compound. In certain embodiments, the pharmaceutical composition comprises 0.5% to 99.5% of the aforementioned compound. In certain embodiments, the pharmaceutical composition comprises 1% to 99% of the aforementioned compound. In certain embodiments, the pharmaceutical composition comprises 2% to 98% of the aforementioned compound.

[0613] In certain embodiments, the pharmaceutical composition contains 0.01%-99.99% of a pharmaceutically acceptable carrier, diluent, or excipient, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1%-99.9% of a pharmaceutically acceptable carrier, diluent, or excipient. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of a pharmaceutically acceptable carrier, diluent, or excipient. In certain embodiments, the pharmaceutical composition contains 1%-99% of a pharmaceutically acceptable carrier, diluent, or excipient. In certain embodiments, the pharmaceutical composition contains 2%-98% of a pharmaceutically acceptable carrier, diluent, or excipient.

[0614] All compounds disclosed herein, as well as mixtures and compositions comprising the compounds disclosed herein, can be administered to a living organism via any route of administration. Routes of administration include oral administration, intravenous injection, intramuscular injection, subcutaneous injection, rectal administration, vaginal administration, sublingual administration, nasal inhalation, oral inhalation, eye drops, and topical or systemic transdermal administration.

[0615] All compounds disclosed herein, as well as mixtures and compositions containing the compounds disclosed herein, can be formulated into single doses containing the active compounds disclosed herein as well as carriers, excipients, and the like. The dosage forms can be tablets, capsules, injections, granules, powders, suppositories, pills, creams, pastes, gels, powders, oral solutions, inhalants, suspensions, dry suspensions, patches, lotions, and the like. These dosage forms can contain ingredients commonly used in pharmaceutical preparations, such as diluents, absorbents, wetting agents, binders, disintegrants, colorants, pH adjusters, antioxidants, antibacterial agents, isotonicity adjusters, and anti-adherent agents.

[0616] Suitable formulations of the above-mentioned various dosage forms are available from public sources, such as Remington: The Science and Practice of Pharmacy, 21st edition, published by Lippincott Williams & Wilkins in 2006 and Rowe, Raymond C. Handbook of Pharmaceutical Excipients, Chicago, Pharmaceutical Press in 2005. Therefore, those skilled in the art can easily prepare them.

[0617] Different dosages can be selected based on the nature and intensity of the disease suffered by different individuals, the patient's age, gender, weight, administration route and other factors. The dosage of the compound of the present disclosure can be 0.01 to 500 mg / kg per day, preferably 1-100 mg / kg per day, and can be administered in a single or multiple doses.

[0618] The present invention also provides the compound of the present invention or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the aforementioned pharmaceutical composition, for use in preventing and / or treating skin-related diseases.

[0619] In certain embodiments, the skin-related disease is selected from diseases related to hyperkeratosis of the skin.

[0620] In certain embodiments, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, pruritus, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis.

[0621] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin, and doxorubicin.

[0622] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel and capecitabine.

[0623] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor, a VEGFR inhibitor, c-kit, a PDGFR, an ALK inhibitor, a RET inhibitor, and a FGFR inhibitor.

[0624] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor and a VEGFR inhibitor.

[0625] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, fruquintinib, anlotinib, erlotinib, afatinib, and osimertinib.

[0626] The present invention also provides the use of the compound of the present invention or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the aforementioned pharmaceutical compositions for preventing and / or treating skin-related diseases.

[0627] In certain embodiments, the skin-related disease is selected from diseases related to hyperkeratosis of the skin.

[0628] In certain embodiments, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, pruritus, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis.

[0629] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin, and doxorubicin.

[0630] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel and capecitabine.

[0631] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor, a VEGFR inhibitor, c-kit, a PDGFR, an ALK inhibitor, a RET inhibitor, and a FGFR inhibitor.

[0632] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor and a VEGFR inhibitor.

[0633] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, fruquintinib, anlotinib, erlotinib, afatinib, and osimertinib.

[0634] The present invention also provides use of the compound of the present invention or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the aforementioned pharmaceutical compositions in the preparation of drugs for preventing and / or treating skin-related diseases.

[0635] In certain embodiments, the skin-related disease is selected from diseases related to hyperkeratosis of the skin.

[0636] In certain embodiments, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, pruritus, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis.

[0637] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin, and doxorubicin.

[0638] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel and capecitabine.

[0639] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor, a VEGFR inhibitor, c-kit, a PDGFR, an ALK inhibitor, a RET inhibitor, and a FGFR inhibitor.

[0640] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor and a VEGFR inhibitor.

[0641] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, fruquintinib, anlotinib, erlotinib, afatinib, and osimertinib.

[0642] The present invention also provides the compound of the present invention or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the aforementioned pharmaceutical composition, which is used to prevent and / or treat diseases related to abnormal RNA m6A methylation.

[0643] In certain embodiments, the disease associated with abnormal RNA m6A methylation is selected from skin-related diseases.

[0644] In certain embodiments, the skin-related disease is selected from diseases related to hyperkeratosis of the skin.

[0645] In certain embodiments, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, pruritus, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis.

[0646] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin, and doxorubicin.

[0647] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel and capecitabine.

[0648] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor, a VEGFR inhibitor, c-kit, a PDGFR, an ALK inhibitor, a RET inhibitor, and a FGFR inhibitor.

[0649] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor and a VEGFR inhibitor.

[0650] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, fruquintinib, anlotinib, erlotinib, afatinib, and osimertinib.

[0651] The present invention also provides the use of the compound of the present invention or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the aforementioned pharmaceutical compositions for preventing and / or treating diseases related to abnormal RNA m6A methylation.

[0652] In certain embodiments, the disease associated with abnormal RNA m6A methylation is selected from skin-related diseases.

[0653] In certain embodiments, the skin-related disease is selected from diseases related to hyperkeratosis of the skin.

[0654] In certain embodiments, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, pruritus, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis.

[0655] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin, and doxorubicin.

[0656] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel and capecitabine.

[0657] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor, a VEGFR inhibitor, c-kit, a PDGFR, an ALK inhibitor, a RET inhibitor, and a FGFR inhibitor.

[0658] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor and a VEGFR inhibitor.

[0659] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, fruquintinib, anlotinib, erlotinib, afatinib, and osimertinib.

[0660] The present invention also provides use of the compound of the present invention or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating diseases related to abnormal RNA m6A methylation.

[0661] In certain embodiments, the disease associated with abnormal RNA m6A methylation is selected from skin-related diseases.

[0662] In certain embodiments, the skin-related disease is selected from diseases related to hyperkeratosis of the skin.

[0663] In certain embodiments, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, pruritus, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis.

[0664] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, adriamycin, oxaliplatin, and doxorubicin.

[0665] In certain embodiments, the hand-foot syndrome is caused by administration of one or more selected from the group consisting of docetaxel and capecitabine.

[0666] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor, a VEGFR inhibitor, c-kit, a PDGFR, an ALK inhibitor, a RET inhibitor, and a FGFR inhibitor.

[0667] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of an EGFR inhibitor and a VEGFR inhibitor.

[0668] In certain embodiments, the hand-foot skin reaction is caused by administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, fruquintinib, anlotinib, erlotinib, afatinib, and osimertinib.

[0669] Explanation of terms:

[0670] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0671] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, Methylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents selected from deuterium atoms, halogen, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0672] The residue derived from the above alkyl group by removing a hydrogen atom from the parent atom is referred to as "alkylene".

[0673] The term "alkenyl" refers to an alkyl compound containing at least one carbon-carbon double bond in the molecule, wherein alkyl is as defined above. Alkenyl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from alkoxy, halogen, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0674] The term "alkynyl" refers to an alkyl compound containing at least one carbon-carbon triple bond in the molecule, wherein alkyl is as defined above. Alkynyl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more groups independently selected from alkoxy, halogen, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0675] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, preferably 3 to 8 (e.g., 3, 4, 5, 6, 7, and 8) carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls.

[0676] The term " spiroalkyl " refers to a polycyclic group of 5 to 20 yuan, a carbon atom (called spiral atom) shared between the monocycle, which can contain one or more double bonds. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan (for example, 7, 8, 9 or 10 yuan). According to the number of spiral atoms shared between the rings, spiroalkyl is divided into single spiroalkyl, double spiroalkyl or multiple spiroalkyl, preferably single spiroalkyl and double spiroalkyl. More preferably, it is 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiroalkyl. Non-limiting examples of spiroalkyl include:

[0677] Term " fused cycloalkyl " refers to 5 to 20 yuan, and each ring in the system shares the full carbon polycyclic group of a pair of carbon atoms adjacent to other rings in the system, and wherein one or more rings can contain one or more double bonds.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan (such as 7,8,9 or 10 yuan).Can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl according to the number of composition ring, preferably bicyclic or tricyclic, more preferably 3 yuan / 4 yuan, 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 4 yuan, 5 yuan / 5 yuan, 5 yuan / 6 yuan, 6 yuan / 3 yuan, 6 yuan / 4 yuan, 6 yuan / 5 yuan and 6 yuan / 6 yuan bicyclic alkyl.Non-limiting examples of fused cycloalkyl include:

[0678] The term "bridged cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, and which may contain one or more double bonds. Preferably, it is 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9, or 10 members). Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl group, preferably a bicyclic, tricyclic, or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged cycloalkyl groups include:

[0679] The cycloalkyl group includes the cycloalkyl group as described above (including monocyclic, spirocyclic, fused and bridged rings) fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, non-limiting examples include etc.; preferred

[0680] The cycloalkyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0681] The term "alkoxy" refers to -O-(alkyl) and -O-(cycloalkyl), wherein alkyl and cycloalkyl are as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, propoxy, and butoxy. Alkoxy may be optionally substituted or unsubstituted, and when substituted, it is preferably one or more of the following groups independently selected from deuterium atoms, halogen, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0682] The term "heterocyclyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic cyclic substituent containing 3 to 20 ring atoms, one or more of which is a heteroatom selected from nitrogen, oxygen and sulfur, wherein the sulfur may be optionally oxoed (i.e., forming a sulfoxide or sulfone), but excluding the ring portion of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12), of which 1 to 4 (e.g., 1, 2, 3 and 4) are heteroatoms; more preferably, it contains 3 to 8 ring atoms (e.g., 3, 4, 5, 6, 7 and 8), of which 1 to 3 (e.g., 1, 2 and 3) are heteroatoms; more preferably, it contains 3 to 6 ring atoms, of which 1 to 3 are heteroatoms; and most preferably, it contains 5 or 6 ring atoms, of which 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, tetrahydropyranyl, 1,2,3,6-tetrahydropyridinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclyls include spiro, fused, and bridged heterocyclyls.

[0683] The term "spiro heterocyclic radical" refers to a non-aromatic polycyclic heterocyclic group of 5 to 20 members, one atom (called spiral atom) shared between the monocycles, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, and the sulfur can be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon. It can contain one or more double bonds. It is preferably 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members). According to the number of shared spiral atoms between the rings, the spiro heterocyclic radical is divided into a monospiro heterocyclic radical, a dispiro heterocyclic radical or a polyspiro heterocyclic radical, preferably a monospiro heterocyclic radical and a dispiro heterocyclic radical. It is more preferably a 3 / 5 member, 3 / 6 member, 4 / 4 member, 4 / 5 member, 4 / 6 member, 5 / 5 member or a 5 / 6 member monospiro heterocyclic radical. Non-limiting examples of spiro heterocyclic radicals include:

[0684] The term "fused heterocyclic radical" refers to 5 to 20 yuan, and each ring in the system shares a pair of adjacent atoms of the non-aromatic polycyclic heterocyclic group with other rings in the system, and one or more rings can contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, and the sulfur can be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 yuan, and more preferably 7 to 10 yuan (e.g., 7, 8, 9 or 10 yuan). According to the number of the rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic radicals, preferably bicyclic or tricyclic, more preferably 3 yuan / 4 yuan, 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 4 yuan, 5 yuan / 5 yuan, 5 yuan / 6 yuan, 6 yuan / 3 yuan, 6 yuan / 4 yuan, 6 yuan / 5 yuan and 6 yuan / 6 yuan bicyclic fused heterocyclic radicals. The limiting examples of fused heterocyclic radicals include:

[0685] The term "bridged heterocyclic group" refers to a 5 to 14-membered, non-aromatic polycyclic heterocyclic group of any two rings sharing two atoms that are not directly connected, which may contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, and the sulfur may be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members). According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

[0686] The heterocyclyl ring includes a heterocyclyl as described above (including monocyclic, spiro heterocyclic, fused heterocyclic and bridged heterocyclic rings) fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclyl, non-limiting examples of which include:

[0687] wait.

[0688] The heterocyclyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0689] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (fused polycyclic is a ring that shares adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. The aryl group includes an aryl ring as described above fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

[0690] The aryl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0691] The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 (e.g., 1, 2, 3, and 4) heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl is preferably 5 to 10 yuan (e.g., 5, 6, 7, 8, 9, or 10 yuan), more preferably 5 or 6 yuan, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, etc. The heteroaryl includes a heteroaryl fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. Non-limiting examples of heteroaryl include:

[0692] The heteroaryl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0693] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0694] The term "hydroxy" refers to -OH.

[0695] The term "amino" refers to -NH2.

[0696] The term "cyano" refers to -CN.

[0697] The term "nitro" refers to -NO2.

[0698] The term "stereoisomers" refers to compounds that have identical chemical constitution but differ in the way the atoms or groups are arranged in space. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric (cis / trans) isomers, atropisomers, and the like.

[0699] The term "tautomer" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine, lactam-lactim isomerizations, pyrazolyl isomerizations, and the like.

[0700] The compounds of the present disclosure include isotopic derivatives thereof. The term "isotopic derivative" refers to a compound whose structure differs only in the presence of one or more isotopically enriched atoms. For example, a compound having a structure of the present disclosure, with "deuterium" or "tritium" replacing hydrogen, or with 18 F-fluorine labeling ( 18 F isotope) instead of fluorine, or with 11 C-,13 C-, or 14 C-enriched carbon ( 11 C-, 13 C-, or 14 C-carbon labeling; 11 C-, 13 C-, or 14 Compounds in which a carbon atom is replaced by a deuterium atom (C-isotope) are within the scope of the present disclosure. Such compounds can be used as analytical tools or probes in, for example, biological assays, or as in vivo diagnostic imaging tracers for diseases, or as tracers for pharmacodynamic, pharmacokinetics, or receptor studies. The various deuterated forms of the compounds disclosed herein refer to compounds in which each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of compounds with reference to relevant literature. Commercially available deuterated starting materials can be used when preparing deuterated compounds, or they can be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated iodoethane, and deuterated iodomethane. Deuterated compounds generally retain activity comparable to undeuterated compounds, and when deuterated at certain specific sites, better metabolic stability can be achieved, thereby obtaining certain therapeutic advantages.

[0701] "Substituted" means that one or more hydrogen atoms, preferably 1 to 5, more preferably 1 to 3 hydrogen atoms, in a group are independently replaced by a corresponding number of substituents. Those skilled in the art will be able to determine (by experiment or theory) whether substitution is possible or not without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0702] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0703] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present disclosure that are safe and effective for use in mammals and possess the desired biological activity. Salts can be prepared during the final isolation and purification of the compound, or separately by reacting a suitable group with a suitable base or acid. Bases commonly used to form pharmaceutically acceptable salts include inorganic bases and organic bases. Acids commonly used to form pharmaceutically acceptable salts include inorganic acids and organic acids.

[0704] With respect to a drug or pharmacologically active agent, the term "therapeutically effective amount" refers to a non-toxic amount of the drug or agent sufficient to achieve the desired effect. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active agent. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

[0705] As used herein, the term "solvate" refers to a physical association of a compound of the present disclosure with one or more, preferably 1-3, solvent molecules, whether organic or inorganic. This physical association includes hydrogen bonding. In some cases, for example, when one or more, preferably 1-3, solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate will be isolated. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Solvation methods are well known in the art.

[0706] "Prodrug" refers to a compound that can be transformed in vivo under physiological conditions, for example, by hydrolysis in the blood, to yield the active prodrug.

[0707] The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with patient tissues without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio, and effective for the intended use.

[0708] As used herein, the singular form "a," "an," and "the" include plural references and vice versa unless the context clearly dictates otherwise.

[0709] The above definitions are given for the terms involved in the present invention. Those skilled in the art can also understand the above terms in combination with the existing technology. The following is a further description based on the content of the present invention and the definitions of the terms.

[0710] The following examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. Experimental methods in the following examples, where specific conditions are not specified, are generally performed under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise indicated, parts and percentages are by weight. In the preparation processes, each reaction is carried out in an inert solvent at room temperature to reflux temperature. The reaction time is generally 0.25 to 48 hours.

[0711] Furthermore, the LC-MS liquid chromatography-mass spectrometry instrument uses a Waters ACQUITY Arc or equivalent equipment. The mass spectrometry (MS) uses an ESI source and only indicates the molecular weight M of the parent molecule, usually reporting [M+H] +NMR spectra were obtained using a Varian 400 MHz nuclear magnetic resonance spectrometer or equivalent, typically using CDCl₃ or DMSO-d₆ as solvents, and chemical shifts are reported in ppm. Peaks are described as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd (doublet of doublets). Coupling constants are expressed in Hz.

[0712] The abbreviations used in the experimental section are listed below: AA ammonium acetate AcOH acetic acid Boc tert-butoxycarbonyl DAST diethylaminosulfur trifluoride DIEA N,N-diisopropylethylamine DCM dichloromethane DMSO dimethyl sulfoxide DMF dimethylamide THF tetrahydrofuran TFA trifluoroacetic acid HATU 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0713] Example 1: Preparation of 6-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 1)

[0714] 3-Bromo-2-oxocyclohexane-1-carboxylic acid ethyl ester (20 mg, 0.08 mmol) and 4-fluoroaniline (22 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1 M HCl and once with saturated 100 mL of NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The mixture was separated and purified by silica gel column chromatography to obtain 6-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylic acid ethyl ester (15.6 mg). 10 mg of ethyl 6-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.038 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried, and concentrated, and then ammonia water, HATU (0.038 mmol), and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 6-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 1, 6.2 mg) as a white solid. LCMS [M+H] + :233. 1 H-NMR (400MHz, CDCl3): δ8.52(s,1H),7.32-7.41(m,1H),7.24-7.30(m,1H),6.76- 6.82(m,1H),5.76-5.81(m,2H),3.68(m,1H),2.67-2.72(m,2H),1.64-1.90(m,4H).

[0715] Example 2: Preparation of 6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 2), (R)-6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 2a), and (S)-6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 2b)

[0716] Ethyl 3-bromo-2-oxocyclohexane-1-carboxylate (20 mg, 0.08 mmol) and 4-chloroaniline (25 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1N HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The mixture was separated and purified by silica gel column chromatography to obtain ethyl 6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (16 mg). 10 mg of ethyl 6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.036 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2 M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried, and concentrated, and then ammonia water, HATU (0.036 mmol), and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (compound 2, 7.6 mg) as a white solid. The (R)-6-chloro-2,3,4,9-tetrahydro-1H-carbazolecarboxamide (compound 2a) and (S)-6-chloro-2,3,4,9-tetrahydro-1H-carbazolecarboxamide (compound 2b) were separated using a Chiralpak AD chiral column. Compounds 2a and 2b LCMS [M+H] + :249. 1 H-NMR (400MHz, DMSO-d6): δ10.79 (s, 1H), 7.37-7.39 (m, 2H), 7.28 (d, J = 8.0Hz, 1H), 7. 08(s,1H),6.98-7.01(m,1H),3.64-3.67(m,1H),2.58-2.61(m,2H),1.66-2.04(m,4H).

[0717] Example 3: Preparation of 6-chloro-5-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 3a)

[0718] Ethyl 3-bromo-2-oxocyclohexane-1-carboxylate (20 mg, 0.08 mmol) and 4-chloro-3-fluoroaniline (29 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1 M HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The product was separated and purified by silica gel column chromatography to obtain ethyl 6-chloro-5-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (10 mg). 10 mg of ethyl 6-chloro-5-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.034 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried, and concentrated, and then ammonia water, HATU (0.036 mmol), and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was purified by preparative liquid chromatography (SFC) to obtain 6-chloro-5-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (compound 3a, 7 mg) as a white solid. LCMS [M+H] + :267. 1 H-NMR (400MHz, CDCl3): δ8.63(s,1H),7.45(d,J=8Hz1H),7.10(d,J=8.0Hz,1H),5 .52-5.68(m,2H),3.75(t,J=6.2Hz,1H),2.71(t,J=5.5Hz,2H),1.66-2.24(m,4H).

[0719] Example 4: Preparation of 6-chloro-7-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 3b)

[0720] Ethyl 3-bromo-2-oxocyclohexane-1-carboxylate (20 mg, 0.08 mmol) and 4-chloro-3-fluoroaniline (29 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1 M HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The product was separated and purified by silica gel column chromatography to obtain ethyl 6-chloro-7-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (9 mg). 9 mg of ethyl 6-chloro-7-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.03 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried, and concentrated, and then ammonia water, HATU (0.036 mmol), and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. 6-chloro-7-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (compound 3b, 6 mg) was obtained as a white solid by SFC preparative liquid phase separation and purification. LCMS [M+H] + :267. 1 H-NMR (400MHz, CDCl3): δ 8.71(s,1H),7.31-7.43(m,1H),7.23(s,1H),5.51-5.71(m,2H),3.62-3.65(m,1H),2.59-2.63(m,2H),1.66-2.01(m,4H).

[0721] Example 5: Preparation of 6-chloro-5,7-difluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 4)

[0722] Ethyl 3-bromo-2-oxocyclohexane-1-carboxylate (20 mg, 0.08 mmol) and 4-chloro-3,5-difluoroaniline (32.7 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1 M HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The mixture was separated and purified by silica gel column chromatography to obtain ethyl 6-chloro-5,7-difluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (16 mg). 10 mg of ethyl 6-chloro-5,7-difluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.032 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried, and concentrated, and then ammonia water, HATU (0.036 mmol), and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 6-chloro-5,7-difluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (compound 4, 6.8 mg) as a white solid. LCMS [M+H] + :285. 1 H-NMR (400MHz, CDCl3): δ8.72(s,1H),7.28-7.35(m,1H),5.35-5.41(m,2H),3.58-3.65(m,1H),2.58-2.65(m,2H),1.67-2.04(m,4H).

[0723] Example 6: Preparation of 2-chloro-5,6,7,8,9,10-hexahydro-7,10-methoxycyclohepta[b]indole-6-carboxamide (Compound 5), (S)-2-chloro-5,6,7,8,9,10-hexahydro-7,10-methoxycyclohepta[b]indole-6-carboxamide (Compound 5a) and (R)-2-chloro-5,6,7,8,9,10-hexahydro-7,10-methoxycyclohepta[b]indole-6-carboxamide (Compound 5b)

[0724] Ethyl 4-bromo-3-oxobicyclo[3.2.1]octane-2-carboxylate (22 mg, 0.08 mmol) and 4-chloroaniline (25 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1M HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The mixture was separated and purified by silica gel chromatography to give ethyl 2-chloro-5,6,7,8,9-hexahydro-7,10-methylcyclohepta[b]indole-6-carboxylate (18.5 mg). 10 mg of 2-chloro-5,6,7,8,9-hexahydro-7,10-methylcyclohepta[b]indole-6-carboxylic acid ethyl ester (0.033 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried and concentrated, and then ammonia water, HATU (0.036 mmol) and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 2-chloro-5,6,7,8,9,10-hexahydro-7,10-methylcyclohepta[b]indole-6-carboxamide (compound 5, 7.3 mg) as a white solid. The product was purified by Chiralpak AD chiral column separation gave (S)-2-chloro-5,6,7,8,9,10-hexahydro-7,10-methanocyclohepta[b]indole-6-carboxamide (compound 5a) and (R)-2-chloro-5,6,7,8,9,10-hexahydro-7,10-methanocyclohepta[b]indole-6-carboxamide (compound 5b). Compounds 5a and 5b LCMS [M+H] + :275. 1 H-NMR (400MHz, DMSO-d6): δ10.74(s,1H),7.30-7.33(m,2H),7.24(d,J=8.0Hz,1H),7.18(s,1H),6 .93(d,J=8.0Hz,1H),3.52-3.58(m,1H),2.77-2.80(m,1H),2.17-2.20(m,1H),1.56-1.92(m,6H).

[0725] Example 7: Preparation of 6-chloro-4-methyl-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 6)

[0726] 3-Bromo-4-methyl-2-oxocyclohexane-1-carboxylic acid ethyl ester (21 mg, 0.08 mmol) and 4-chloroaniline (25 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1M HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The mixture was separated and purified by silica gel column chromatography to obtain 6-chloro-4-methyl-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylic acid ethyl ester (18 mg). 10 mg of ethyl 6-chloro-4-methyl-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.034 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried and concentrated, and then ammonia water, HATU (0.036 mmol) and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 6-chloro-4-methyl-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (compound 6, 7.1 mg) as a white solid. LCMS [M+H] + :263. 1 H-NMR (400MHz, CDCl3): δ8.72-8.77(m,1H),8.44(s,1H),7.54-7.56(m,1H),7.45(s,1H),7.21-7.23(m,2H),7.09-7.12(m,2 H),5.62-5.75(m,4H),3.71(s,1H),3.37(m,1H),3.10-3.12(m,1H),2.72-2.73(m,1H),2.03-2.27(m,4H),1.19-1.37(m,6H).

[0727] Example 8: Preparation of 6-chloro-4-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 7)

[0728] Ethyl 3-bromo-4-fluoro-2-oxocyclohexane-1-carboxylate (21.4 mg, 0.08 mmol) and 4-chloroaniline (25 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1 M HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The product was separated and purified by silica gel column chromatography to give ethyl 6-chloro-4-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (19 mg). 10 mg of ethyl 6-chloro-4-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.034 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried and concentrated, and then ammonia water, HATU (0.036 mmol) and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 6-chloro-4-fluoro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (compound 7, 7.3 mg) as a white solid. LCMS [M+H] + :267. 1 H-NMR (400MHz, CDCl3): δ8.53(s,1H),7.36-7.39(m,1H),7.28-7.31(m,1H),7.10-7.15(m, 1H), 5.64-5.78 (m, 2H), 5.51 (td, J = 51.6, 4.1Hz, 1H), 3.63-3.66 (m, 1H), 1.63-1.95 (m, 4H).

[0729] Example 9: Preparation of 8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxamide (Compound 8), (R)-8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxamide (Compound 8a) and (S)-8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxamide (Compound 8b)

[0730] Ethyl 2-bromo-1-methyl-3-oxopiperidine-4-carboxylate (21.1 mg, 0.08 mmol) and 4-chloroaniline (25 mg, 0.2 mmol) were mixed and heated to 150°C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed three times with 100 mL of 1M HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The mixture was separated and purified by silica gel column chromatography to give ethyl 8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxylate (16.8 mg). 10 mg of 8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxylic acid ethyl ester (0.036 mmol) was dissolved in 10 mL of ethanol, and 2 mL of 2M LiOH solution was added. The mixture was stirred at room temperature for 1 hour. After rotary distillation, 20 mL of water was added to dilute the mixture and the pH was adjusted to 2. The mixture was then extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried and concentrated, and then ammonia water, HATU (0.036 mmol) and acetonitrile (5 mL) were added. The mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxamide (compound 8, 6.8 mg) as a white solid. The product was purified by Chiralpak AD chiral column separation gave (R)-8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxamide (Compound 8a) and (S)-8-chloro-1-methyl-2,3,4,5-tetrahydropyrido[3,2-b]indole-4-carboxamide (Compound 8b). Compounds 8a and 8b LCMS [M+H] + :264. 1 H-NMR (400MHz, DMSO-d6): δ10.90(s,1H),7.59(s,1H),7.33-7.37(m,3H),6.92(d,J =8.0Hz,1H),3.60-3.63(m,1H),2.96-3.03(m,2H),2.79(s,3H),1.92-2.10(m,2H).

[0731] Example 10: Preparation of 6-chloro-7-phenylamino-2,3,4,9-tetrahydro-1H-carbazolecarboxamide (Compound 9)

[0732] Ethyl 3-bromo-2-oxocyclohexane-1-carboxylate (1.3 g, 6.07 mmol), 6-chloro-1N-benzene-1,3-diamine (0.5 g, 2.02 mmol) and ethylene glycol (1 ml) were heated to 110°C and reacted for 2 h. The mixture was quenched with water and extracted with ethyl acetate. The organic phase was desolvated under reduced pressure and subjected to column chromatography to obtain 0.22 g of crude product 9-1. 9-1 (0.15 g, 0.406 mmol, 1.0 eq), 0.6 ml (4V) of a THF / MeOH / H2O (4:4:1) mixture, and LiOH (97.5 mg, 4.07 mmol, 10.0 eq) were taken and stirred at room temperature overnight, then concentrated under reduced pressure. Ethyl acetate (2 ml) and water (2 ml) were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine, dried, and concentrated under reduced pressure. Ammonia water, HATU (0.44 mmol), and acetonitrile (5 mL) were added to the residue, and the mixture was reacted at room temperature for 24 hours. The mixture was separated and purified by silica gel column chromatography to obtain 6-chloro-7-phenylamino-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (compound 9, 6.9 mg) as a white solid. LCMS [M+H] + :340. 1 H-NMR (400MHz, CD3OD): δ7.38(s,1H),7.16-7.23(m,3H),6.94-6.96(m,2H), 6.78-6.82(m,1H),3.64-3.72(m,1H),2.73-2.77(m,2H),1.68-2.10(m,4H).

[0733] Example 11: Preparation of 3-fluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide (Compound 10)

[0734] (2-Bromophenyl)hydrazine hydrochloride (7.83 g, 35.03 mmol), 4-hydroxycyclohexane-1-one (4 g, 35.03 mmol), and acetic acid (80 mL) were added to a reaction flask, the atmosphere was replaced with nitrogen, and the reaction was continued at 110°C for 2 hours. The mixture was concentrated and diluted with dichloromethane. The mixture was extracted with saturated sodium bicarbonate solution, and the organic phase was washed with saturated brine. The organic phase was concentrated and purified by silica gel column chromatography to obtain 8-bromo-2,3,4,9-tetrahydro-1H-carbazol-3-yl acetate (7.04 g). 8-Bromo-2,3,4,9-tetrahydro-1H-carbazol-3-yl acetate (7.04 g), methanol (30 mL), and a 30% aqueous sodium hydroxide solution (10 mL) were added to the reaction flask and stirred at 50°C for 1 hour. The reaction was complete after TLC monitoring. The methanol was removed by rotary evaporation under reduced pressure, and the mixture was extracted three times with ethyl acetate. The organic phase was concentrated. 8-Bromo-2,3,4,9-tetrahydro-1H-carbazole-3-ol (5.09 g) was purified by silica gel column chromatography. 8-Bromo-2,3,4,9-tetrahydro-1H-carbazole-3-ol (5.09 g) and dichloromethane (50 mL) were added to a reaction flask and stirred until dissolved. The mixture was replaced with nitrogen and cooled to -10°C. DAST (1.21 g) was added and reacted at -10°C for 1.5 hours. The reaction solution was quenched by adding dropwise to an ice-saturated sodium bicarbonate solution and extracted with dichloromethane. The organic phase was concentrated and purified by silica gel column chromatography to give 8-bromo-3-fluoro-2,3,4,9-tetrahydro-1H-carbazole (758 mg). 8-Bromo-3-fluoro-2,3,4,9-tetrahydro-1H-carbazole (758 mg), DMF (10 mL), Pd(dppf)Cl2 (414 mg, 0.566 mmol), and triethylamine (859 mg, 8.49 mmol) were placed in a reaction flask, replaced with CO, and reacted at 80°C for 16 hours. The reaction was monitored by TLC. The reaction was cooled to room temperature, and ethyl acetate (30 mL) was added to dilute the reaction solution. The reaction solution was washed three times with saturated brine, and the organic phase was concentrated and purified by silica gel column chromatography to obtain 3-fluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxylic acid (78 mg). 3-Fluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxylic acid (78 mg), ammonia (24 mg), TCFH (140 mg), NMI (55 mg), and acetonitrile (5 mL) were placed in a reaction flask, replaced with nitrogen, and stirred at room temperature for 2 hours. The reaction solution was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by preparative liquid phase to obtain 0.915 mg of 3-fluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide (Compound 10). LCMS [M+H] + :233. 1H-NMR (400MHz, CDCl3): δ7.97(s,1H),7.32-7.41(m,1H),7.24-7.30(m,1H),6.96-7.00(m,1H),5.76-5.8 1(m,2H),5.09-5.26(m,1H),2.95-3.16(m,3H),2.81-2.88(m,1H),2.26-2.34(m,1H),2.06-2.21(m,1H).

[0735] Example 12: Synthesis of 3,3-difluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide (Compound 11)

[0736] (2-Bromophenyl)hydrazine hydrochloride (1 g, 4.474 mmol), 4,4-difluorocyclohexane-1-one (0.6 g, 4.474 mmol) and acetic acid (10 mL) were added to a reaction flask, replaced with nitrogen, stirred at 110 ° C for 2 hours, concentrated the reaction solution, diluted with dichloromethane, and extracted with saturated sodium bicarbonate aqueous solution. The organic phase was washed with saturated brine, concentrated, and purified by silica gel column chromatography to give 8-bromo-3,3-difluoro-2,3,4,9-tetrahydro-1H-carbazole (991 mg). 8-Bromo-3,3-difluoro-2,3,4,9-tetrahydro-1H-carbazole (891 mg), DMF (10 mL), Pd(dppf)Cl2 (456 mg, 0.623 mmol), and triethylamine (946 mg, 9.34 mmol) were placed in a reaction flask, replaced with CO, and reacted at 100°C for 4 hours. The reaction was monitored by TLC. The reaction was cooled to room temperature, and ethyl acetate (30 mL) was added to dilute the reaction solution. The reaction solution was washed three times with saturated brine, and the organic phase was concentrated and purified by silica gel column chromatography to obtain 3,3-difluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxylic acid (57 mg). 3,3-Difluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxylic acid (57 mg), ammonia (16 mg), TCFH (96 mg), NMI (38 mg) and acetonitrile (5 mL) were placed in a reaction flask, replaced with nitrogen, and stirred at room temperature for 2 hours. The reaction solution was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by preparative liquid phase to obtain 3,3-difluoro-2,3,4,9-tetrahydro-1H-carbazole-8-carboxamide (10 mg). LCMS [M+H] + :251. 1H NMR(400MHz,DMSO-d6)δ11.31(s,1H),8.09(br.s.,1H),7.53(br.s.,1H),7.38-7.40(m,1H),7.28-7.29 (m,1H),6.91-7.00(m,1H),3.51(t,J=14.4Hz,2H),2.89(t,J=6.6Hz,2H),2.23(tt,J=13.9,6.7Hz,2H).

[0737] Example 13: 5-chloro-9,12-diazatetracycle (10.2.1.0 2,10 .0 3,8 ) Synthesis of Pentadeca-2(10),3,5,7-tetraene-11-carboxamide (Compound 12)

[0738] 5-Chloro-1H-indole (25 g, 164.9 mmol), 1H-pyrrole-2,5-dione (24 g, 247.35 mmol), and acetic acid (150 mL) were added to a reaction flask, the atmosphere was purged with nitrogen, and the mixture was reacted at 120°C for 48 hours. The reaction solution was concentrated and purified by silica gel column chromatography to obtain 3-(5-chloro-1H-indol-3-yl)pyrrolidine-2,5-dione (6 g). Lithium aluminum hydride (4.554 g, 120 mmol) and tetrahydrofuran (120 mL) were placed in a reaction flask, the atmosphere was purged with nitrogen, the temperature was lowered to 0°C, and 3-(5-chloro-1H-indol-3-yl)pyrrolidine-2,5-dione (6 g, 24 mmol) was added portionwise. The mixture was stirred for 10 minutes, then the temperature was raised to reflux and the reaction was allowed to react for 16 hours. The reaction was then cooled to 0°C and quenched by the addition of water followed by a 10% aqueous solution of NaOH. The mixture was stirred at room temperature for 4 hours, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain 5-chloro-3-(pyrrolidin-3-yl)-1H-indole (4.6 g). 5-chloro-3-(pyrrolidin-3-yl)-1H-indole (4.6 g, 20.84 mmol), ethanol (46 mL) and a toluene solution of ethyl glyoxylate (50%) (5.1 g, 25 mmol) were added to a microwave reaction tube, nitrogen was replaced, and microwave reaction was carried out at 95°C for 30 minutes. The reaction solution was concentrated and purified by silica gel column chromatography to obtain 5-chloro-9,12-diazatetracyclic (10.2.1.0 2,10 .0 3,8 ) ethyl pentadecan-2(10),3,5,7-tetraene-11carboxylate (500 mg). Add 5-chloro-9,12-diazatetracyclo (10.2.1.0 2,10 .0 3,8) ethyl pentadecan-2(10),3,5,7-tetraene-11carboxylate (450 mg, 1.477 mmol), tetrahydrofuran (10 mL), methanol (10 mL), lithium hydroxide (353 mg) and water (5 mL) were reacted at room temperature for 12 hours. The reaction was completed after monitoring by TLC. The reaction solution was concentrated, 3M hydrochloric acid was added to adjust the pH value to 4-5, and ethyl acetate was added for extraction. The organic phase was concentrated and purified by silica gel column chromatography to obtain 5-chloro-9,12-diazatetracyclic (10.2.1.0 2,10 .0 3,8 )pentadecan-2(10),3,5,7-tetraene-11carboxylic acid (56 mg). 5-Chloro-9,12-diazatetracyclo (10.2.1.0 2,10 .0 3,8 )pentadecan-2(10),3,5,7-tetraene-11carboxylic acid (56 mg, 0.202 mmol), acetonitrile (2 mL), DMF (1 mL), NH4Cl (21.6 mg, 0.404 mmol) and DIPEA (78.3 mg, 0.606 mmol), replaced with nitrogen, cooled to 0 ° C, added with HATU (153.6 mg, 0.404 mmol), stirred for 4 h, concentrated the reaction solution, purified by silica gel column chromatography to obtain a crude product, and then purified by preparative HPLC to obtain 5-chloro-9,12-diazatetracyclic (10.2.1.0 2,10 .0 3,8 )pentadecan-2(10),3,5,7-tetraene-11carboxamide (1.6 mg). LCMS [M+H] + :275. 1 H-NMR (400MHz, CD3OD): δ8.39 (br, 1H), 7.48-7.50 (m, 1H), 7.26-7.32 (m, 1H), 7.03-7.06 (m, 1H), 4.63 ( s, 1H), 3.49-3.52 (m, 2H), 3.34-3.36 (m, 1H), 2.98-3.08 (m, 2H), 2.17-2.24 (m, 1H), 2.03-2.09 (m, 1H).

[0739] Example 14: Drug-induced changes in keratinocyte mRNA methylation levels

[0740] 1*10 8HaCat cells were plated in culture dishes and treated with 50 nM of each drug (sorafenib, capecitabine, docetaxel, and osimertinib). The cells were cultured at 37°C in a 5% CO2 incubator for 24 hours. The culture medium was removed, the cells were washed with PBS, centrifuged, and the supernatant discarded. 400 μL of RLT cell lysis buffer (QIAGEN, Cat. #79216) was added to the cells. A portion of the sample was added with 400 μL of 70% ethanol, stirred thoroughly, and transferred to an RNeasy mini column. RNA was extracted using the RNeasy mini kit (QIAGEN, Cat. #74104). 2 μL of RNA sample was dissolved in 98 μL of 10 mM Tris-HCl buffer, and the absorbance at 260 nm was measured using a Nanodrop (Thermo Fisher Scientific) to determine RNA concentration.

[0741] 40 μg of RNA sample was taken and mRNA (PolyA+RNA) was enriched using the NEBNext Poly(A) mRNA Magnetic Separation Module (NEB, Cat.#E7490). The 3' end of the enriched mRNA (PolyA+RNA) molecule was then connected to the reverse transcription adapter RTA using the Direct RNA Sequencing Kit (Oxfod Nanopore Technologies, Cat.#SQK-RNA002). The mRNA (PolyA+RNA) molecule was used as a template for reverse transcription reaction to synthesize its complementary chain. The reverse transcription adapter RTA end was connected to the sequencing adapter (RNA adapter) to form the final sequencing library, and the library was sequenced using Qubit TM dsDNA HS assay kit (Invitrogen, Cat.# LOT2133187) was used for quantitative detection.

[0742] After library quality control, the prepared sequencing library was loaded onto a PromethION Flow Cell chip (Oxford Nanopore Technologies, Cat.# FLO-MIN106D) and sequenced using a PromethION sequencer (Oxford Nanopore Technologies) in the matching sequencing mode. Sequencing data were base-called using guppy, and m6A methylation analysis was performed using the original Fast5 data.

[0743] The results, as shown in Figure 1, show that compared to the blank control group, the m6A methylation levels of HaCat cells increased by 5.05-fold, 6.01-fold, 3.97-fold, and 5.13-fold after addition of sorafenib, capecitabine, docetaxel, and osimertinib, respectively. This suggests that anti-cancer drugs (sorafenib, capecitabine, docetaxel, and osimertinib) can induce aberrant m6A methylation in human keratinocyte mRNA. Further bioinformatics analysis revealed significant differences in the m6A methylation levels of NAP1L2, primarily manifested in m6A methylation of the UGAGGACUCA fragment.

[0744] Example 15: Changes in mRNA methylation levels in a drug-induced mouse skin adverse reaction model

[0745] ICR male mice (5-6 weeks old, weighing approximately 35 grams) were acclimated for 7 days and then randomly divided into 6 groups: blank control, sorafenib, capecitabine, docetaxel, and osimertinib. Each group was gavaged with the corresponding modeling drug (sorafenib 100 mg / kg, capecitabine 200 mg / kg, docetaxel 25 mg / kg, osimertinib 10 mg / kg) once daily for 30 consecutive days before being sacrificed. Skin tissue from the hind limbs and toes of the mice was collected. A small amount of tissue sample was ground into a powder in a mortar filled with liquid nitrogen. The single-phase lysis buffer was added and the sample was allowed to stand at room temperature for 5 minutes before centrifugation (12,000 rpm) for 5 minutes. 1 mL of the supernatant was removed, 200 μL of chloroform was added, and the mixture was shaken and allowed to stand at room temperature for 15 minutes. The sample was then centrifuged (12,000 rpm, 4°C) for 15 minutes. The upper aqueous phase was aspirated and an equal volume of isopropanol was added. The sample was allowed to stand at -20°C for 1 hour and then centrifuged (12,000 rpm, 4°C). The supernatant was discarded. 400 μL of 70% ethanol was added to the pellet, mixed thoroughly, and transferred to an RNeasy mini column. RNA was extracted using the RNeasy mini kit (QIAGEN, Cat. #74104). 2 μL of RNA sample was dissolved in 98 μL of 10 mM Tris-HCl buffer, and the absorbance at 260 nm was measured using a Nanodrop (Thermo Fisher Scientific) to determine RNA concentration.

[0746] 40 μg of RNA sample was taken and mRNA (PolyA+RNA) was enriched using the NEBNext Poly(A) mRNA Magnetic Separation Module (NEB, Cat.#E7490). The 3' end of the enriched mRNA (PolyA+RNA) molecule was then connected to the reverse transcription adapter RTA using the Direct RNA Sequencing Kit (Oxfod Nanopore Technologies, Cat.#SQK-RNA002). The mRNA (PolyA+RNA) molecule was used as a template for reverse transcription reaction to synthesize its complementary chain. The reverse transcription adapter RTA end was connected to the sequencing adapter (RNA adapter) to form the final sequencing library, and the library was sequenced using Qubit TM dsDNA HS assay kit (Invitrogen, Cat.# LOT2133187) was used for quantitative detection.

[0747] After library quality control, the prepared sequencing library was loaded onto a PromethION Flow Cell chip (Oxford Nanopore Technologies, Cat.# FLO-MIN106D) and sequenced using a PromethION sequencer (Oxford Nanopore Technologies) in the matching sequencing mode. Sequencing data were base-called using guppy, and m6A methylation analysis was performed using the original Fast5 data.

[0748] The results are shown in Figure 2. Compared with the positive control group, the m6A methylation level in the skin tissue of the hind limbs and toes of mice in the drug modeling group was significantly increased. The m6A methylation level of NAP1L2 in the drug-treated group was significantly different from that in the blank control group.

[0749] Example 16: m6A regulates the mRNA expression level of NAP1L2

[0750] 1*10 8HaCat cells were plated in culture dishes and treated with recombinant m6A methylase METTL3 protein (Abcam, Cat.#ab271611) or m6A demethylase FTO protein (Abcam, Cat.#ab271525), or transfected with siRNA for METTL3 (sh-METTL3, GeneChip) or siRNA for FTO (sh-FTO, GeneChip). The cells were cultured at 37°C in an incubator containing 5% carbon dioxide for 24 hours, washed with PBS, and collected by high-speed centrifugation. Total RNA was extracted using Trizol reagent (Sigma Aldrich). 1 μg of RNA was reverse-transcribed into cDNA using a cDNA reverse transcription kit (Transgene Biotech, Cat. #AT311-03). RT-PCR was performed by adding 1.25 μL of primers (Beyotime, Cat. #QH18721S), 10 μL of iTag Universal SYBR Green supermix (Bio-Rad, Cat. #172-5125), and an appropriate amount of DEPC ultrapure water to a 20 μL reaction mixture. After completion of the reaction, the reaction mixture was subjected to agarose gel electrophoresis to determine the expression level of NAP1L2 mRNA.

[0751] The results are shown in Figure 3. When the m6A methylase METTL3 or the m6A methylase FTO function was inhibited (sh-FTO) in keratinocytes HaCat, the mRNA expression of NAP1L2 increased significantly; when the m6A methylase FTO or the m6A methylase METTL3 function was inhibited (sh-METTL3) in keratinocytes HaCat, the mRNA expression of NAP1L2 decreased significantly. This indicates that m6A methylation can regulate the mRNA expression of NAP1L2.

[0752] Example 17: NAP1L2 regulates the expression of matrix metal proteins in human keratinocytes HaCat

[0753] 1*10 8 HaCat cells were plated in culture dishes and treated with recombinant human NAP1L2 protein (Abcam, Cat.#ab117213), sorafenib, capecitabine, docetaxel, and osimertinib, or transfected with NAP1L2 siRNA (sh-NAP1L2, GeneChip). The cells were incubated at 37°C in a 5% CO2 incubator for 24 hours. After washing with PBS, the cells were harvested by high-speed centrifugation. RLT lysis buffer was added to the harvested cells, shaken at 4°C for half an hour, and the supernatant was collected after centrifugation for measurement of matrix metal protein expression by Western blotting.

[0754] Total RNA was extracted from the collected cells using Trizol reagent (Sigma Aldrich). 1 μg of RNA was reverse-transcribed into cDNA using a cDNA reverse transcription kit (Transgene Biotech, Cat. #AT311-03). RT-PCR was performed by adding 1.25 μL of primers (SinoBiological, Cat. #HP100168 & HP100367), 10 μL of iTag Universal SYBR Green supermix (Bio-Rad, Cat. #172-5125), and an appropriate amount of DEPC ultrapure water to a 20 μL reaction solution. Following completion of the reaction, the reaction solution was subjected to agarose gel electrophoresis to determine the mRNA expression level of the metal matrix protein.

[0755] The results are shown in Figure 4. When NAP1L2 recombinant protein was added to keratinocytes HaCat, the expression levels of the mRNA and protein of the metalloproteinases MMP2 and MMP9 increased significantly. When NAP1L2 siRNA was added to keratinocytes HaCat to inhibit the expression of NAP1L2, the expression levels of the mRNA and protein of the metalloproteinases MMP2 and MMP9 decreased significantly. Many studies have confirmed that abnormal expression of metalloproteinases is closely related to various skin-related diseases (Kumper M. et al., Am. J. Physiol. Cell. Physiol. 2022, 323(4): 1290-1303).

[0756] Example 18: m6A regulates the expression of human keratinocyte differentiation markers

[0757] 1*10 8 HaCat cells were plated in culture dishes and incubated with recombinant HBEGF protein (Thermo Fisher Scientific, Cat.#100-47-1mg) at 37°C in an incubator containing 5% CO2 for 24 hours. HBEGF antibody (Invitrogen, Cat.#406316) or transfection with siRNA for METTL3 (sh-METTL3, GeneChip) or NAP1L2 (sh-NAP1L2, GeneChip) were then added and incubated at 37°C in an incubator containing 5% CO2 for 24 hours. After washing with PBS, the cells were harvested by high-speed centrifugation. RLT lysis buffer was added to the harvested cells, shaken at 4°C for half an hour, and the supernatant was collected after centrifugation. Protein expression of the keratinocyte differentiation markers KRT1, KRT10, Loricrin, and Involucrin was measured by Western blotting.

[0758] Total RNA was extracted using Trizol reagent (Sigma Aldrich), and 1 μg of RNA was reverse transcribed into cDNA using a cDNA reverse transcription kit (Transgene Biotech, Cat. #AT311-03). RT-PCR was performed by adding 1.25 μL of primers (primer sequences are shown in Table 2), 10 μL of iTag Universal SYBR Green supermix (Bio-Rad, Cat. #172-5125), and an appropriate amount of DEPC ultrapure water to a 20 μL reaction solution. After completion of the reaction, the reaction solution was subjected to agarose gel electrophoresis to measure the mRNA expression levels of the keratinocyte differentiation markers KRT1, KRT10, loricrin, and involucrin.

[0759] The results are shown in Figure 5. When HBEGF recombinant protein was added to keratinocytes HaCat, the mRNA and protein expression levels of keratinocyte differentiation markers KRT1, KRT10, Loricrin, and Involucrin significantly increased, indicating that the keratinocytes HaCat were in a highly differentiated state. However, when HBEGF antibodies were added or siRNAs targeting METTL3 and NAP1L2 were used to inhibit intracellular m6A methylation, the mRNA and protein expression levels of keratinocyte differentiation markers KRT1, KRT10, Loricrin, and Involucrin significantly decreased. This suggests that m6A can regulate the expression of keratinocyte differentiation markers, thereby regulating keratinocyte differentiation.

[0760] Example 19: Effects of Compounds on m6A Methylation Levels in THP-1 Cells

[0761] 1*10 8THP-1 cells were seeded in culture dishes and treated with 400 nM compound 1, 4 nM and 400 nM compounds 2b, 5a, and 8b, 4 nM and 4 μM nicotinamide, and 5 μM of the m6A methylase inhibitor UZH2. The cells were cultured at 37°C in a 5% CO2 incubator for 24 hours. The medium was discarded, the cells were washed with PBS, and the supernatant was discarded by centrifugation. 400 μL of RLT cell lysis buffer was added and the cells were stored at -80°C. A portion of the sample was taken, 400 μL of 70% ethanol was added, and the mixture was mixed thoroughly. 700 μL of the sample was transferred to an RNeasy spin column and centrifuged for 15 seconds (8000 g, 25°C). 700 μL of RW1 buffer was added and centrifuged for 15 seconds (8000 g, 25°C). Finally, 500 μL of RPE buffer was added and centrifuged for 16 seconds (8000 g, 25°C). This step was repeated twice, followed by centrifugation for 2 minutes to completely remove the eluate. Add 30 μL of nuclease-free water to the column, incubate for 5 minutes, and centrifuge for 2 minutes (12,000 g, 25°C). Repeat this step three times. Dissolve 2 μL of RNA sample in 98 μL of 10 mM Tris buffer and measure the absorbance at 260 nm using a Nanodrop to determine the RNA concentration.

[0762] Dissolve 50 μg of total RNA in 100 μL of nuclease-free water. Resuspend the Oligo(dT) magnetic beads, transfer 50 μL of beads to a 1.5 mL tube, add 500 μL of binding buffer, let stand, and remove the supernatant. Add 100 μL of RNA sample to 200 μL of magnetic bead suspension, mix, and incubate in a Theromixer at 25°C for 5 minutes. Discard the supernatant and add 200 μL of wash buffer. Repeat twice, then centrifuge for 10 seconds (2000 g, 25°C). Add 50 μL of elution buffer and mix thoroughly. Heat to 75°C, transfer the supernatant to a new 1.5 mL tube, and purify using the RNA Clean & Concentrator kit to determine the RNA concentration.

[0763] Add 20 μL (about 200 ng) of mRNA sample and 20 μL of Nuclease P1 digestion premix (0.5 μL of 2 unit / μL Nuclease P1, 0.4 μL of 5 M NaCl, 2 μL of 0.1 M ZnCl2 and 17.1 μL of PCR-grade water) to each test tube, stir and incubate at 37°C in a Theromixer for 2 hours, add 2 μL of 2M NH4HCO3 solution and 1 unit of alkaline phosphatase, stir and incubate at 37°C in a Theromixer for 2 hours, add 1 μL of 1.2 M HCl neutralization solution, centrifuge for 30 minutes (16000g, 4°C), take 20 μL of supernatant and analyze m6A methylation fragments by LC-MS, and calculate the m6A inhibition rate.

[0764] m6A inhibition rate (%) = (measured ion peak area - blank control) / (positive ion peak area - blank control) * 100

[0765] The results, as shown in Figure 6, show that compounds 2b, 5a, and 8b significantly inhibited m6A methylation levels in cellular mRNA in a concentration-dependent manner. At a concentration of 400 nM, compound 2b exhibited comparable m6A inhibitory activity to the m6A methylase inhibitor UZH2 at a concentration of 5 μM. However, nicotinamide had no inhibitory effect on m6A methylation levels in cellular mRNA at either low (4 nM) or high (4 μM) concentrations.

[0766] Example 20: Effects of compounds on m6A methylation levels in human keratinocytes

[0767] HaCat cells were cultured in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin. The culture dishes were placed in an incubator containing 5% CO2 and cultured at 37°C for 24 hours. An equal volume of DMSO was added to control group 1, and 50 nM sorafenib, capecitabine, docetaxel, and osimertinib were added to control groups A, B, C, and D, respectively. Sample groups A1-17, B1-17, C1-17, and D1-17 were added with 400 nM of the test compound or nicotinamide after adding 50 nM of the corresponding compound. The cells were cultured continuously for 24 hours, washed with PBS, and collected by high-speed centrifugation. Total RNA was extracted using Trizol reagent (Sigma Aldrich), and m6A methylation in the sample RNA was quantitatively detected using the EpiQuik m6A RNA Methylation Quantification Kit. The standard curve used positive control samples at concentrations of 0.01 ng / μl, 0.02 ng / μl, 0.05 ng / μl, 0.1 ng / μl, 0.2 ng / μl, and 0.5 ng / μl. The absorbance at 450 nm was read using a Tecan GENios microplate reader. The m6A quantitative calculation formula is as follows:

[0768] m6A (ng) = (absorbance of sample well – absorbance of background well) / slope of standard curve

[0769] m6A (%) = m6A (ng) / sample RNA amount (ng) * 100%

[0770] The results are shown in Table 1. Treatment with sorafenib, capecitabine, docetaxel, and osimertinib resulted in a significant increase in m6A methylation levels in human keratinocytes (HaCat). Compounds 2, 2b, 5a, and 8b effectively inhibited the abnormal increase in RNA m6A levels in human keratinocytes induced by chemotherapy drugs and multikinase inhibitors.

[0771] Table 1. Fold change of m6A methylation level relative to control group 1. Calculation method: Fold change = measured m6A amount / measured m6A amount in control group 1.

[0772] Example 21: Inhibitory effect of compounds on keratinocyte HaCaT differentiation

[0773] Human keratinocytes HaCaT were cultured in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin. 6 Cells were plated in 96-well plates at a density of 10 cells per square centimeter and incubated at 37°C in a 5% CO2 incubator for 24 hours. An equal volume of DMSO was added to control well 1, and 2.5 ng / mL human recombinant HBEGF protein (Abcam, ab205523) was added to control well 2. Sample wells were then incubated for 24 hours. After washing with PBS, cells were harvested by high-speed centrifugation. Total RNA was extracted using Trizol reagent (Sigma Aldrich). 1 μg of RNA was reverse-transcribed into cDNA using a cDNA reverse transcription kit (Transgene Biotech, AT311-03). RT-PCR reactions were performed by adding 1.25 μL of primers (primer sequences are shown in Table 2), 10 μL of iTag Universal SYBR Green Supermix (Bio-Rad, 172-5125), and an appropriate amount of DEPC ultrapure water to a 20 μL reaction mixture. After the reaction, the reaction solution was subjected to agarose gel electrophoresis to determine the mRNA expression levels of keratinocyte differentiation markers KRT1, KRT10, Loricrin, and Involucrin.

[0774] Table 2. Some primer sequence information

[0775] As shown in Figure 7, after induction with human recombinant HBEGF protein, the mRNA expression levels of HaCaT differentiation markers KRT1, KRT10, Loricrin, and Involucrin significantly increased. However, after the addition of some compounds (such as compounds 2, 2b, 3a, 3b, 5a, and 8b), the mRNA expression levels of HaCaT differentiation markers KRT1, KRT10, Loricrin, and Involucrin were significantly suppressed.

[0776] Example 22: Inhibitory effect of the compound on keratinocyte differentiation in the rat hand-foot skin reaction model

[0777] After SD rats (weighing approximately 200 g) were bred and adapted for one week, they were divided into groups according to their weight, with 12 rats in each group. The modeling drugs (sorafenib, erlotinib, afatinib and osimertinib) were dissolved in solutions containing 5% DMSO, 45% PEG400 and 50% H2O, respectively, and the modeling drugs were diluted to the required concentrations and administered orally once a day according to the doses shown in Table 3. One hour after oral administration, 0.05 g of ointment containing different mass ratios of the test compound was evenly applied to the left paw of the rat, while the right paw was applied with a blank matrix ointment as a self-control. The rats in the blank control group were not applied with the drug. After applying the drug, the limbs were fixed for 2 hours, and then the residual drug was wiped off with clean water, the fixation was released and free movement was restored. The modeling drug, blank matrix ointment, and test compound ointment were all administered once daily for 30 consecutive days. The rats were euthanized, and paw plantar skin tissue was obtained, fixed in 10% neutral formaldehyde, cut into 5 μm sections, dehydrated, and embedded in paraffin. The test compound ointment was prepared by mixing the compound, white beeswax, white petrolatum, and light liquid paraffin in specific ratios (weight ratios of the three ointments were: 1:18:58:23, 3:18:57:22, and 10:20:40:30, respectively). The blank matrix ointment was prepared by mixing white beeswax, white petrolatum, and light liquid paraffin in specific ratios (weight ratio: 18:60:22).

[0778] Tissue staining: After dewaxing and rehydrating, the paw and plantar skin tissue sections of the above rats were stained in a hematoxylin solution for several minutes. After washing, the sections were immersed in 1% acidic alcohol until the sections faded to a light blue-red color. After washing with running water for 5 minutes, the sections were stained with eosin for 2-3 minutes. After washing to remove excess dye, the sections were dehydrated and transparentized in xylene for several minutes. The sections were then sealed with a neutral resin. The stratum corneum was observed under an optical microscope, and the thickness of the epidermal stratum corneum was measured using Dmetrix software.

[0779] Immunohistochemical staining: After dewaxing and hydration, the paw plantar skin tissue sections of the above rats were incubated with 3% H2O2 at room temperature for 30 minutes. After antigen retrieval, they were blocked with 10% goat serum for 30 minutes. KRT1 antibodies (Abcam, ab93652) and Loricrin antibodies (Abcam, ab183646) were added dropwise and incubated overnight at 4°C. HRP secondary antibodies (ZSGB-BIO, PV-6001) and DAB kits (ZSG-BIO, ZLI9017) were added for color development. The sections were counterstained with hematoxylin, washed, and mounted with central resin. The expression levels of KRT1 and Loricrin were observed under a light microscope.

[0780] The criteria for successful rat model establishment are: (i) the appearance of symptoms such as erythema, swelling, desquamation, ulceration, or blisters on the paw; and / or (ii) a significantly higher stratum corneum thickness than that of normal rats as determined by tissue staining; and / or (iii) a significant increase in markers such as KRT1, KRT5, and Loricrin. The incidence rate was calculated as the proportion of animals in each group meeting the above criteria for successful model establishment: incidence rate = (number of rats with successful model establishment / total number of rats in the group) * 100%.

[0781] The rat tissue staining pathology scoring criteria are as follows: no blisters, 0 points; 1-3 blisters, 1 point; 4-6 blisters, 2 points; 7-9 blisters, 3 points; and more than 10 blisters, 4 points. Inflammation occupying less than 10% of the total section area is scored as 0 points; inflammation occupying 10-25% of the total section area is scored as 1 point; inflammation occupying 25-50% of the total section area is scored as 2 points; inflammation occupying 50-75% of the total section area is scored as 3 points; and inflammation occupying more than 75% of the total section area is scored as 4 points. A score of 0 was assigned if the hyperemia area accounted for less than 10% of the total slice area; a score of 1 was assigned if the hyperemia area accounted for 10-25% of the total slice area; a score of 2 was assigned if the hyperemia area accounted for 25-50% of the total slice area; a score of 3 was assigned if the hyperemia area accounted for 50-75% of the total slice area; and a score of 4 was assigned if the hyperemia area accounted for more than 75% of the total slice area. Scoring was performed using a double-blind scoring system, and data from each group were presented as mean ± SEM (N = 12).

[0782] Table 3. Drug dosage and experimental results in rat models

[0783] As shown in Table 3, sorafenib, erlotinib, afatinib, and osimertinib were administered orally once daily in SD rats, with success rates of establishing hand-foot skin reaction models of 75%, 83.3%, 66.7%, and 75%, respectively. Some compounds (e.g., 2, 2b, 5a, and 8b) significantly reduced the incidence of hand-foot skin reaction at the application site at low doses (e.g., 1% compound content). Some compounds (e.g., 2, 2b, 5a, and 8b) significantly reduced the incidence of hand-foot skin reaction at all paws of rats at high concentrations (e.g., 3% and 10% compound content).

[0784] As shown in Figure 8, the stratum corneum thickness measurements of rats successfully treated with sorafenib, erlotinib, afatinib, and osimertinib showed significantly higher stratum corneum thickness in the paw and plantar skin than in normal rats. Some compounds (e.g., 2, 2b, 5a, and 8b) were able to effectively reduce stratum corneum thickness in the paw and plantar skin of rats undergoing the treatment.

[0785] As shown in Figure 9, the tissue staining results show that drug-induced skin toxicity can cause blister formation in the subcutaneous and epidermal regions of rat paws, inflammatory cell infiltration in the dermis, and congestion in the skin tissue. Compounds 2b and 5a effectively ameliorated drug-induced subcutaneous and epidermal blister formation, inhibited inflammatory cell infiltration in the dermis, and significantly improved skin congestion.

[0786] As shown in Figure 10, the pathological scoring results indicate that the drug-induced blister formation in the subcutaneous and epidermal regions of the rat paws, inflammatory cell infiltration in the dermis, and skin congestion. Compounds 2, 2b, 5a, and 8b effectively ameliorated the drug-induced blister formation in the subcutaneous and epidermal regions, inhibited inflammatory cell infiltration in the dermis, and significantly improved skin congestion.

[0787] Immunohistochemical staining also showed that KRT1 and Loricrin expression increased significantly in rat tissues after successful treatment with sorafenib, erlotinib, afatinib, and osimertinib. However, with the addition of the compounds, KRT1 and Loricrin expression decreased significantly. In summary, compounds 2, 2b, 5a, and 8b can effectively inhibit keratinocyte differentiation in vitro and in vivo, demonstrating potential for the treatment of hyperkeratosis-related diseases.

[0788] Example 23: Effect of Compound 2b in a Chemotherapy-Induced Mouse Hand-Foot Syndrome Model

[0789] After 7 days of adaptive feeding, SPF ICR mice (male, 5-6 weeks old, weighing approximately 35 grams) were randomly divided into the following groups: blank control group (6 mice), capecitabine model group (6 mice), docetaxel model group (6 mice), capecitabine + compound 2b group (6 mice), docetaxel + compound 2b group (6 mice), capecitabine + nicotinamide group (6 mice), and docetaxel + nicotinamide group (6 mice). Each group was given the corresponding modeling drug (capecitabine 200 mg / kg, docetaxel 25 mg / kg) by oral gavage once a day; the blank control group was given an equal volume of normal saline. After 2 weeks of chemotherapy drug administration, each group was topically applied with the corresponding compound ointment used in Example 22 (compound content 3%) or orally administered with nicotinamide (100 mg / kg) once a day for 16 consecutive days.

[0790] Measurement of toe swelling: The degree of toe swelling of the mouse hind limbs was measured using a toe swelling tester (KW-7C, Nanjing Calvin Biotechnology Co., Ltd.) before administration of the modeling drug, 2 weeks after administration of the modeling drug, and before sacrifice. The same position was marked on the joint of the mouse hind foot before administration, 2 weeks after administration, and before sacrifice. During measurement, the water was exactly on the same horizontal line as the marked position, and the value was recorded after it stabilized.

[0791] Skin histopathological staining: Skin tissue samples from the hind limbs of mice were fixed in 4% paraformaldehyde at room temperature for 4 hours. The tissues were then removed and rinsed with running water for several hours. After dehydration with 70%, 80%, and 90% ethanol solutions, they were treated with a mixture of equal parts pure alcohol and xylene for 15 minutes. Permeabilization was performed twice with xylene for 15 minutes each, until the sample became transparent. The samples were then immersed in a mixture of 50% xylene and 50% paraffin for 15 minutes, followed by permeabilization with Paraffin I and Paraffin II for one hour each. After paraffin embedding, the samples were sliced, baked, dewaxed, and hydrated. The hydrated sections were then stained in hematoxylin solution (Zhongshan Jinqiao, Catalog No. 23041001) for 3 minutes, differentiated in hydrochloric acid ethanol solution for 15 seconds, washed with water, and blued in Scott blue solution (Servicebio, Catalog No. 20230801) for 15 seconds. After rinsing with running water, the sections were stained with eosin stain (Solarbio, Catalog No. 33535) for 3 minutes. After rinsing with running water, the sections were dehydrated, transparentized, mounted, and examined under a microscope.

[0792] Immunohistochemical staining: Paraffin sections of mouse hind limb skin tissue were baked, dewaxed, and hydrated, followed by antigen retrieval using 0.2 M citric acid buffer (1.534 g citric acid, 3.1 g sodium citrate, 100 mL distilled water, pH 4.7). Endogenous hydrogen peroxide was removed with 3% hydrogen peroxide (Shandong Anjie High-Tech Disinfection Technology Co., Ltd., Catalog No. 20290902). After incubation at room temperature for 10 minutes, sections were rinsed thoroughly with PBS buffer (8 g NaCl, 0.2 g KCl, 1.44 g Na₂HPO₄, 1 M HCl, pH 7.4). Sections were permeabilized with 0.5% Triron-X-100 (Aimejie Technology, Catalog No. A-CSH436-100 mL). Mouse anti-IL-1β and rabbit anti-IL-8 were permeabilized for 10 minutes each, while rabbit anti-IL-6 was not permeabilized. After blocking with 5% BSA antigen, a 1 / 150 dilution of mouse anti-IL-1β (Affinity Biosciences, Catalog No. BF8021), a 1 / 200 dilution of rabbit anti-IL-8 (Affinity Biosciences, Catalog No. DF6998), or a 1 / 150 dilution of rabbit anti-IL-6 (Affinity Biosciences, Catalog No. DF6087) was added to each slide and incubated in a humidified chamber at 4°C overnight. Remove the wet chamber after overnight incubation and let it stand at room temperature for 45 minutes. Wash the slides three times with PBS buffer for 15 minutes each. Add a 1 / 100 dilution of horseradish enzyme-conjugated goat anti-rabbit IgG (H+L) (Zhongshan Jinqiao, Cat. No. 234750811) to rabbit anti-IL-8 and rabbit anti-IL-6 markers. Add a 1 / 100 dilution of horseradish enzyme-conjugated goat anti-mouse IgG (H+L) (Zhongshan Jinqiao, Cat. No. 226700804) to mouse anti-IL-1β markers. Incubate at 37°C for 30 minutes and then rinse thoroughly with PBS buffer. Add DAB (CWBIO, Cat. No. 13723) for color development for 3-5 minutes, wash with PBS buffer for 1 minute, counterstain with hematoxylin (Zhongshan Jinqiao, Cat. No. 23041001) for 3 minutes, differentiate with hydrochloric acid and alcohol to turn blue, and wash with water. The sections were sequentially placed in 80% alcohol, 95% alcohol, anhydrous ethanol, and anhydrous ethanol II for rapid dehydration, and then transparentized with xylene I and xylene II. The sections were then sealed with neutral resin glue (Solarbio, product number 20230725) and examined under a microscope.

[0793] Figure 11 shows the skin appearance of mice. Fourteen days after the capecitabine and docetaxel treatment, significant lesions appeared on the skin of the hind limbs and toes of the mice. Compared with the normal control group, the capecitabine and docetaxel groups showed significant redness, swelling, cracking, and blisters on the skin of the hind limbs and toes of the mice.

[0794] Figure 12 shows the extent of hind paw swelling in mice. Following 14 consecutive days of oral administration of the modeling drugs capecitabine and docetaxel, hind paw swelling was observed in mice. Topical application of compound 2b significantly improved drug-induced paw swelling in mice (p < 0.05). However, oral administration of 100 mg / kg nicotinamide did not significantly improve hind paw swelling in mice.

[0795] The results of pathological staining of the toe skin tissue are shown in Figure 13. Compared with the normal control group, the modeling drugs capecitabine and docetaxel, after 14 consecutive days of oral administration, caused significant thickening of the epidermis and stratum corneum in the skin tissue of the hind limbs and toes of mice, an increase in basal granular cells, and visible inflammatory cell infiltration. Topical application of compound 2b to the hind limbs and toes of mice can effectively improve the thickening of the epidermis and stratum corneum of the skin tissue, and the morphology of the basal cells returned to normal, with no inflammatory cell infiltration. However, oral administration of 100 mg / kg nicotinamide slightly improved the thickening of the epidermis and stratum corneum of the skin tissue of the hind limbs and toes of mice, with a slightly increased number of basal granular cells and still visible inflammatory cell infiltration.

[0796] The immunohistochemical results of the capecitabine modeling series are shown in Figure 14. Compared with the normal control group, the modeling drug capecitabine caused a significant increase in the cytokine IL-8 in the skin tissue of the mouse hind limbs and toes after 14 consecutive days of oral administration (p<0.05). This shows that oral administration of capecitabine can cause skin inflammation in mice, which is consistent with the results of clinical trial observations. Topical application of compound 2b to the hind limbs and toes of mice can significantly reduce the expression level of the cytokine IL-8 in the skin tissue of the mouse hind limbs and toes (p<0.05). However, oral administration of 100 mg / kg nicotinamide had no significant effect on the expression level of the cytokine IL-8 in the skin tissue of the mouse hind limbs and toes.

[0797] The immunohistochemical results of the docetaxel modeling series are shown in Figure 15. Compared with the normal control group, the modeling drug docetaxel caused a significant increase in the cytokine IL-6 in the skin tissue of the mouse hind limbs and toes after 14 consecutive days of oral administration (p < 0.05). This shows that oral administration of docetaxel can cause skin inflammation in mice, which is consistent with the results of clinical trial observations. Topical application of compound 2b to the hind limbs and toes of mice can significantly reduce the expression level of the cytokine IL-6 in the skin tissue of the mouse hind limbs and toes (p < 0.05). However, oral administration of 100 mg / kg nicotinamide had no significant effect on the expression level of the cytokine IL-6 in the skin tissue of the mouse hind limbs and toes.

[0798] Example 24: Effect of Compound 2b in a kinase inhibitor-induced mouse hand-foot skin reaction model

[0799] SPF ICR mice (male, 5-6 weeks old, weighing approximately 35 grams) were randomly divided into the following groups after 7 days of adaptive feeding: blank control group (6 mice), sorafenib modeling group (6 mice), osimertinib modeling group (6 mice), sorafenib + compound 2b group (6 mice), osimertinib + compound 2b group (6 mice), sorafenib + nicotinamide group (6 mice), and osimertinib + nicotinamide group (6 mice). Each group was given the corresponding modeling drug (sorafenib 100 mg / kg, osimertinib 10 mg / kg) by oral gavage once a day; the blank control group was given an equal volume of normal saline. After 2 weeks of chemotherapy drug administration, each group was topically applied with the corresponding compound ointment used in Example 22 (compound content 3%) or orally administered with nicotinamide (100 mg / kg) once a day for 16 consecutive days.

[0800] Measurement of toe swelling: The degree of toe swelling of the mouse hind limbs was measured using a toe swelling tester (KW-7C, Nanjing Calvin Biotechnology Co., Ltd.) before administration of the modeling drug, 2 weeks after administration of the modeling drug, and before sacrifice. The same position was marked on the joint of the mouse hind foot before administration, 2 weeks after administration, and before sacrifice. During measurement, the water was exactly on the same horizontal line as the marked position, and the value was recorded after it stabilized.

[0801] Skin histopathological staining: Skin tissue samples from the hind limbs of mice were fixed in 4% paraformaldehyde at room temperature for 4 hours. The tissues were then removed and rinsed with running water for several hours. After dehydration with 70%, 80%, and 90% ethanol solutions, they were treated with a mixture of equal parts pure alcohol and xylene for 15 minutes. Permeabilization was performed twice with xylene for 15 minutes each, until the sample became transparent. The samples were then immersed in a mixture of 50% xylene and 50% paraffin for 15 minutes, followed by permeabilization with Paraffin I and Paraffin II for one hour each. After paraffin embedding, the samples were sliced, baked, dewaxed, and hydrated. The hydrated sections were then stained in hematoxylin solution (Zhongshan Jinqiao, Catalog No. 23041001) for 3 minutes, differentiated in hydrochloric acid ethanol solution for 15 seconds, washed with water, and blued in Scott blue solution (Servicebio, Catalog No. 20230801) for 15 seconds. After rinsing with running water, the sections were stained with eosin stain (Solarbio, Catalog No. 33535) for 3 minutes. After rinsing with running water, the sections were dehydrated, transparentized, mounted, and examined under a microscope.

[0802] Immunohistochemical staining: Paraffin sections of mouse hind limb skin tissue were baked, dewaxed, and hydrated, followed by antigen retrieval using 0.2 M citric acid buffer (1.534 g citric acid, 3.1 g sodium citrate, 100 mL distilled water, pH 4.7). Endogenous hydrogen peroxide was removed with 3% hydrogen peroxide (Shandong Anjie High-Tech Disinfection Technology Co., Ltd., Catalog No. 20290902). After incubation at room temperature for 10 minutes, sections were rinsed thoroughly with PBS buffer (8 g NaCl, 0.2 g KCl, 1.44 g Na₂HPO₄, 1 M HCl, pH 7.4). Sections were permeabilized with 0.5% Triron-X-100 (Aimejie Technology, Catalog No. A-CSH436-100 mL). Mouse anti-IL-1β and rabbit anti-IL-8 were permeabilized for 10 minutes each, while rabbit anti-IL-6 was not permeabilized. After blocking with 5% BSA antigen, a 1 / 150 dilution of mouse anti-IL-1β (Affinity Biosciences, Catalog No. BF8021), a 1 / 200 dilution of rabbit anti-IL-8 (Affinity Biosciences, Catalog No. DF6998), or a 1 / 150 dilution of rabbit anti-IL-6 (Affinity Biosciences, Catalog No. DF6087) was added to each slide and incubated in a humidified chamber at 4°C overnight. Remove the wet chamber after overnight incubation and let it stand at room temperature for 45 minutes. Wash the slides three times with PBS buffer for 15 minutes each. Add a 1 / 100 dilution of horseradish enzyme-conjugated goat anti-rabbit IgG (H+L) (Zhongshan Jinqiao, Cat. No. 234750811) to rabbit anti-IL-8 and rabbit anti-IL-6 markers. Add a 1 / 100 dilution of horseradish enzyme-conjugated goat anti-mouse IgG (H+L) (Zhongshan Jinqiao, Cat. No. 226700804) to mouse anti-IL-1β markers. Incubate at 37°C for 30 minutes and then rinse thoroughly with PBS buffer. Add DAB (CWBIO, Cat. No. 13723) for color development for 3-5 minutes, wash with PBS buffer for 1 minute, counterstain with hematoxylin (Zhongshan Jinqiao, Cat. No. 23041001) for 3 minutes, differentiate with hydrochloric acid and alcohol to turn blue, and wash with water. The sections were sequentially placed in 80% alcohol, 95% alcohol, anhydrous ethanol, and anhydrous ethanol II for rapid dehydration, and then transparentized with xylene I and xylene II. The sections were then sealed with neutral resin glue (Solarbio, product number 20230725) and examined under a microscope.

[0803] The results, as shown in Figure 16, showed that 14 days after the sorafenib and osimertinib treatments, the skin of the hind limbs and toes of the mice developed significant lesions. Compared with the normal control group, the skin of the hind limbs and toes of the mice in the sorafenib and osimertinib groups showed significant redness, swelling, and cracking.

[0804] Figure 17 shows the extent of hind paw swelling in mice. The modeling drugs sorafenib and osimertinib caused significant hind paw swelling after 14 consecutive days of oral administration. Topical application of compound 2b to the paws of mice effectively alleviated drug-induced paw swelling. However, oral administration of 100 mg / kg nicotinamide did not significantly improve hind paw swelling in mice.

[0805] The results of pathological staining of the toe skin tissue are shown in Figure 18. Compared with the normal control group, the modeling drugs sorafenib and osimertinib, after 14 consecutive days of oral administration, caused significant thickening of the epidermis and stratum corneum in the skin tissue of the mouse hind limb toes, an increase in basal granular cells, and visible inflammatory cell infiltration. Topical application of compound 2b to the mouse hind limb toes can effectively improve the thickening of the epidermis and stratum corneum of the skin tissue, and the morphology of the basal layer cells returned to normal, with no inflammatory cell infiltration. However, oral administration of 100 mg / kg nicotinamide did not significantly improve the thickening of the epidermis and stratum corneum of the mouse hind limb toe skin tissue, and the morphology of the basal layer cells slightly recovered, but inflammatory cell infiltration was still visible.

[0806] The immunohistochemical results of the Sorafenib modeling series are shown in Figure 19. Compared with the normal control group, the modeling drug Sorafenib caused a significant increase in the cytokine IL-1β in the skin tissue of the mouse hind limbs and toes after 14 consecutive days of oral administration (p<0.05). This shows that oral administration of Sorafenib can cause skin inflammation in mice, which is consistent with the results of clinical trial observations. Topical application of compound 2b to the hind limbs and toes of mice can significantly reduce the expression level of the cytokine IL-1β in the skin tissue of the mouse hind limbs and toes (p<0.05). However, oral administration of 100 mg / kg nicotinamide had no significant effect on the expression level of the cytokine IL-1β in the skin tissue of the mouse hind limbs and toes.

[0807] The immunohistochemical results of the osimertinib modeling series are shown in Figure 20. Compared with the normal control group, the modeling drug osimertinib caused a significant increase in the cytokine IL-1β in the skin tissue of the mouse hind limbs and toes after 14 consecutive days of oral administration (p<0.05). This indicates that oral administration of osimertinib can cause skin inflammation in mice, which is consistent with the results of clinical trial observations. Topical application of compound 2b to the hind limbs and toes of mice can significantly reduce the expression level of the cytokine IL-1β in the skin tissue of the mouse hind limbs and toes (p<0.05). However, oral administration of 100mg / kg nicotinamide had no significant effect on the expression level of the cytokine IL-1β in the skin tissue of the mouse hind limbs and toes.

[0808] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and to the same extent as if it were set forth in its entirety herein.

[0809] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations of the claims. Various modifications and variations may be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments may be arbitrarily combined to form additional embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments merely illustrate several implementations of the present invention and do not limit the scope of protection of the patent of this invention.

Claims

1. A compound of formula I or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts: in: X is selected from NR a , O and S; Y is selected from C and N; R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 1 replaced by; R 2 Selected from hydrogen, deuterium, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 2 replaced by; R 3 Selected from hydrogen, deuterium, hydroxyl, -NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 3 replaced by; R 4 Selected from hydrogen, deuterium, halogen, hydroxyl, cyano, -OR b 、-SR c 、-NR i R j , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 4 replaced by; R a is selected from hydrogen, deuterium, alkyl, and haloalkyl; Every R b Each is independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 5 replaced by; Every R c Each is independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 6 replaced by; Every R i , R j are each independently selected from hydrogen, deuterium, -S(O)2-R d , substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 7 replaced by; Every R d Each is independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 8 replaced by; Every R e Each is independently selected from hydrogen, deuterium, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; the substitution is by one or more Q 9 replaced by; Q 1 , Q 2 , Q 3 , Q 4 , Q 5 , Q 6 , Q 7 , Q 8 , Q 9 Each is independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, mercapto, amino, substituted or unsubstituted alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl and heterocyclic; the substitution is by one or more Q 10 replaced by; Q 10 is selected from the group consisting of hydrogen, deuterium, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl; m and n are each independently 0, 1 or 2; p and q are each independently 0, 1, 2, 3 or 4.

2. The compound of formula I according to claim 1 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The compound of formula I is selected from the following structures:

3. The compound of formula I according to claim 1 or 2, or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The X is selected from NR a and O; preferably selected from NR a .

4. The compound of formula I according to any one of claims 1 to 3 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R a is selected from hydrogen, deuterium and alkyl; preferably from hydrogen and C 1-3 Alkyl; more preferably selected from hydrogen and methyl.

5. The compound of formula I according to any one of claims 1 to 4 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R 1 Selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl; preferably selected from hydrogen, deuterium, halogen, -OR b 、-NR i R j , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl and 3-10 membered heterocyclyl; more preferably selected from hydrogen, deuterium, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclic group; most preferably selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , substituted or unsubstituted methyl, cyclopropyl.

6. The compound of formula I according to any one of claims 1 to 5 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The Q 1 is selected from hydrogen, halogen, cyano, hydroxyl, amino, substituted or unsubstituted alkyl; preferably selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl; more preferably selected from hydrogen, halogen; most preferably selected from hydrogen.

7. A compound of formula I according to any one of claims 1 to 6 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R b is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl; preferably from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl; more preferably substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl; most preferably selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

8. The compound of formula I according to any one of claims 1 to 7 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The Q 5 is selected from hydrogen, halogen, cyano, hydroxyl, amino, substituted or unsubstituted heteroaryl; preferably selected from hydrogen, halogen, substituted or unsubstituted 5-10 membered heteroaryl; more preferably selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; and most preferably selected from hydrogen and substituted or unsubstituted pyridyl.

9. The compound of formula I according to any one of claims 1 to 8 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R c is selected from hydrogen, substituted or unsubstituted alkyl and cycloalkyl; preferably from substituted or unsubstituted C 1-6 Alkyl and C 3-7 Cycloalkyl; more preferably substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl; most preferably selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl.

10. The compound of formula I according to any one of claims 1 to 9 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The Q 6 is selected from hydrogen, halogen, cyano, hydroxyl, amino, substituted or unsubstituted heteroaryl; preferably selected from hydrogen, halogen, substituted or unsubstituted 5-10 membered heteroaryl; more preferably selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; and most preferably selected from hydrogen and substituted or unsubstituted pyridyl.

11. The compound of formula I according to any one of claims 1 to 10, or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl; preferably selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 Aryl and 5-10 membered heteroaryl; more preferably selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the most preferred is selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

12. The compound of formula I according to any one of claims 1 to 11 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl; preferably selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 Aryl and 5-10 membered heteroaryl; more preferably selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the most preferred is selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl.

13. The compound of formula I according to any one of claims 1 to 12, or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The Q 7 Selected from hydrogen, halogen, cyano, hydroxyl, amino, substituted or unsubstituted heteroaryl; preferably selected from hydrogen, halogen, substituted or unsubstituted 5-10 membered heteroaryl; more preferably selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, said heteroaryl containing 1-4 heteroatoms selected from N, O and S; most preferably selected from hydrogen and pyridyl.

14. The compound of formula I according to any one of claims 1 to 13 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R d is selected from hydrogen, substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl; preferably selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl, C 6-10 Aryl and 5-10 membered heteroaryl; more preferably selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl and C 6-10 Aryl; most preferably selected from methyl, cyclopropyl and phenyl.

15. The compound of formula I according to any one of claims 1 to 14 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The Q 8 Selected from hydrogen, halogen, cyano, hydroxyl, mercapto and amino; preferably selected from hydrogen, halogen and amino; more preferably selected from hydrogen, fluorine and chlorine; most preferably selected from hydrogen.

16. The compound of formula I according to any one of claims 1 to 15, or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e 、-NR i R j , substituted or unsubstituted alkyl, cycloalkyl, aryl and heteroaryl; preferably selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e , substituted or unsubstituted C 1-6 Alkyl, C 3-7 Cycloalkyl; more preferably selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e , C 1-3 Alkyl, C 3-6 Cycloalkyl; most preferably selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl.

17. A compound of formula I according to any one of claims 1 to 16, or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof, characterized in that: The Q 2 Selected from hydrogen, halogen, cyano, hydroxyl, mercapto and amino; preferably selected from hydrogen, halogen and amino; more preferably selected from hydrogen, fluorine and chlorine; most preferably selected from hydrogen.

18. The compound of formula I according to any one of claims 1 to 17, or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The R e is selected from hydrogen, substituted or unsubstituted alkyl, aryl and heterocyclic groups; preferably selected from hydrogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 Aryl and 3-10 membered heterocyclic group; more preferably selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 Aryl and 3-6 membered heterocyclic group; most preferably selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl.

19. The compound of formula I according to any one of claims 1 to 18, or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The Q 9 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, aryl and heteroaryl; preferably selected from hydrogen, halogen, substituted or unsubstituted C 1-6 Alkyl, C 6-10 Aryl and 5-10 membered heteroaryl; more preferably selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl and 5-6 membered heteroaryl, the heteroaryl contains 1-4 heteroatoms, the heteroatoms are selected from N, O and S; the most preferred is selected from hydrogen, fluorine, methyl, pyrrolyl.

20. The compound of formula I according to any one of claims 1 to 19 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The Q 10 is selected from hydrogen, halogen, substituted or unsubstituted alkyl; preferably selected from hydrogen and halogen; more preferably selected from hydrogen and fluorine.

21. A compound of formula I according to any one of claims 1 to 20 or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof, characterized in that: The R 3 Selected from hydrogen, hydroxyl, -NR i R j , substituted or unsubstituted alkyl and alkoxy; preferably selected from hydrogen, hydroxy, -NR i R j ; More preferably -NR i R j .

22. A compound of formula I according to any one of claims 1 to 21 or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof, characterized in that: The Q 3 Selected from hydrogen, halogen, cyano, hydroxyl, mercapto and amino; preferably selected from hydrogen, halogen and amino; more preferably selected from hydrogen, fluorine and chlorine; most preferably selected from hydrogen.

23. A compound of formula I according to any one of claims 1 to 22, or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof, characterized in that: The R 4 Selected from hydrogen, halogen, -OR b and-NR i R j ; preferably selected from hydrogen and halogen; more preferably selected from hydrogen, fluorine, chlorine and bromine; most preferably selected from hydrogen and fluorine.

24. A compound of formula I according to any one of claims 1 to 23 or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof, characterized in that: The Q 4 is selected from hydrogen, halogen, cyano, hydroxyl, amino, substituted or unsubstituted alkyl; preferably selected from hydrogen, halogen, substituted or unsubstituted C 1-3 Alkyl; more preferably selected from hydrogen, halogen; most preferably selected from hydrogen.

25. A compound of formula I according to any one of claims 1 to 24 or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof, characterized in that: The m is 0 or 1.

26. A compound of formula I according to any one of claims 1 to 25, or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof, characterized in that: The n is 1 or 2.

27. A compound of formula I according to any one of claims 1 to 26, or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof, characterized in that: The p is 1, 2 or 3.

28. A compound of formula I according to any one of claims 1 to 27, or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof, characterized in that: The q is 0, 1 or 2.

29. A compound of formula I according to any one of claims 1 to 28, or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative, and pharmaceutically acceptable salt thereof, characterized in that: The X is selected from NR a ; The R a Selected from hydrogen and C 1-3 alkyl; The R 1 Selected from hydrogen, fluorine, chlorine, bromine, -OR b 、-NR i R j , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl; the substitution is by one or more Q 1 replaced by; The Q 1 is selected from hydrogen and halogen; The R b Selected from substituted or unsubstituted C 1-3 Alkyl and C 3-6 Cycloalkyl; said substitution is by one or more Q 5 replaced by; The Q 5 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by; The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, the heteroaryl containing 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by; The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, C 6-10 Aryl and 5-6 membered heteroaryl, the heteroaryl containing 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 7 replaced by; The Q 7 is selected from hydrogen, halogen, substituted or unsubstituted 5-6 membered heteroaryl, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by; The R d is selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 3-5 Cycloalkyl and C 6-10 Aryl; said substitution is by one or more Q 8 replaced by; The Q 8 is selected from hydrogen, fluorine and chlorine; The R 2 Selected from hydrogen, halogen, -S(O)2-R d 、-C(O)-R e , C 1-3 Alkyl, C 3-6 Cycloalkyl; The R e is selected from hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 6-10 Aryl and 3-6 membered heterocyclic group; the substitution is by one or more Q 9 replaced by; The Q 9 is selected from hydrogen, halogen, substituted or unsubstituted C 1-3 alkyl and 5-6 membered heteroaryl, the heteroaryl containing 1-4 heteroatoms selected from N, O and S; the substitution is by one or more Q 10 replaced by; The Q 10 is selected from hydrogen and halogen; The R 3 Selected from -NR i R j ; The R 4 is selected from the group consisting of hydrogen, fluorine, chlorine and bromine; The m is 0 or 1; The n is 1 or 2; Said p is 1, 2 or 3; The q is 0, 1 or 2.

30. A compound of formula I according to any one of claims 1 to 29 or a stereoisomer, tautomer, solvate, hydrate, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof, characterized in that: The X is selected from NR a ; The R a is selected from hydrogen and methyl; The R 1 Selected from hydrogen, fluorine, chlorine, -OR b 、-NR i R j , methyl, cyclopropyl; The R b is selected from substituted or unsubstituted methyl, cyclopropyl and cyclohexyl; the substitution is by one or more Q 5 replaced by; The Q 5 is selected from hydrogen and substituted or unsubstituted pyridyl; said substitution is by one or more Q 10 replaced by; The R i Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by; The R j Selected from hydrogen, -S(O)2-R d , substituted or unsubstituted methyl, ethyl, cyclopentyl, phenyl, pyridyl, furyl, imidazolyl; the substitution is by one or more Q 7 replaced by; The Q 7 is selected from the group consisting of hydrogen and pyridinyl; The R d is selected from methyl, cyclopropyl and phenyl; The R 2 Selected from hydrogen, fluorine, -S(O)2-R d 、-C(O)-R e , methyl, isopropyl, cyclopropyl; The R e is selected from hydrogen, substituted or unsubstituted methyl, ethyl, phenyl, piperidinyl; the substitution is by one or more Q 9 replaced by; The Q 9 is selected from hydrogen, fluorine, methyl, pyrrolyl; The Q 10 is selected from hydrogen and fluorine; The R 3 Selected from -NR i R j ; The R 4 is selected from hydrogen and fluorine; The m is 0 or 1; The n is 1 or 2; Said p is 1, 2 or 3; The q is 0, 1 or 2.

31. A compound or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, characterized in that: The compound is any of the following:

32. A pharmaceutical composition comprising the compound of any one of claims 1-31 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts.

33. The compound according to any one of claims 1 to 31 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the pharmaceutical composition according to claim 32, for use in preventing and / or treating skin-related diseases; Preferably, the skin-related disease is selected from diseases related to skin hyperkeratosis; Preferably, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, itching, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis; Preferably, the hand-foot syndrome is caused by the administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin and doxorubicin, preferably caused by the administration of one or more selected from the group consisting of docetaxel and capecitabine; Preferably, the hand-foot skin reaction is caused by the administration of one or more of the group consisting of EGFR inhibitors, VEGFR inhibitors, c-kit, PDGFR, ALK inhibitors, RET inhibitors and FGFR inhibitors, preferably caused by the administration of one or more of the group consisting of EGFR inhibitors and VEGFR inhibitors, preferably caused by the administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, furiquintinib, anlotinib, erlotinib, afatinib and osimertinib.

34. Use of the compound according to any one of claims 1 to 31 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the pharmaceutical composition according to claim 32 for preventing and / or treating skin-related diseases; Preferably, the skin-related disease is selected from diseases related to skin hyperkeratosis; Preferably, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, itching, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis; Preferably, the hand-foot syndrome is caused by the administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin and doxorubicin, preferably caused by the administration of one or more selected from the group consisting of docetaxel and capecitabine; Preferably, the hand-foot skin reaction is caused by the administration of one or more of the group consisting of EGFR inhibitors, VEGFR inhibitors, c-kit, PDGFR, ALK inhibitors, RET inhibitors and FGFR inhibitors, preferably caused by the administration of one or more of the group consisting of EGFR inhibitors and VEGFR inhibitors, preferably caused by the administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, furiquintinib, anlotinib, erlotinib, afatinib and osimertinib.

35. Use of the compound according to any one of claims 1 to 31 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the pharmaceutical composition according to claim 32 in the preparation of a medicament for preventing and / or treating skin-related diseases; Preferably, the skin-related disease is selected from diseases related to skin hyperkeratosis; Preferably, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, itching, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis; Preferably, the hand-foot syndrome is caused by the administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin and doxorubicin, preferably caused by the administration of one or more selected from the group consisting of docetaxel and capecitabine; Preferably, the hand-foot skin reaction is caused by the administration of one or more of the group consisting of EGFR inhibitors, VEGFR inhibitors, c-kit, PDGFR, ALK inhibitors, RET inhibitors and FGFR inhibitors, preferably caused by the administration of one or more of the group consisting of EGFR inhibitors and VEGFR inhibitors, preferably caused by the administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, furiquintinib, anlotinib, erlotinib, afatinib and osimertinib.

36. A compound according to any one of claims 1 to 31 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or a pharmaceutical composition according to claim 32, for use in preventing and / or treating diseases associated with abnormal methylation of RNA m6A; Preferably, the RNA m6A abnormal methylation-related disease is selected from skin-related diseases; Preferably, the skin-related disease is selected from diseases related to skin hyperkeratosis; Preferably, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, itching, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis; Preferably, the hand-foot syndrome is caused by the administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin and doxorubicin, preferably caused by the administration of one or more selected from the group consisting of docetaxel and capecitabine; Preferably, the hand-foot skin reaction is caused by the administration of one or more of the group consisting of EGFR inhibitors, VEGFR inhibitors, c-kit, PDGFR, ALK inhibitors, RET inhibitors and FGFR inhibitors, preferably caused by the administration of one or more of the group consisting of EGFR inhibitors and VEGFR inhibitors, preferably caused by the administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, furiquintinib, anlotinib, erlotinib, afatinib and osimertinib.

37. Use of the compound according to any one of claims 1 to 31 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the pharmaceutical composition according to claim 32 for preventing and / or treating diseases related to abnormal methylation of RNA m6A; Preferably, the RNA m6A abnormal methylation-related disease is selected from skin-related diseases; Preferably, the skin-related disease is selected from diseases related to skin hyperkeratosis; Preferably, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, itching, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis; Preferably, the hand-foot syndrome is caused by the administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin and doxorubicin, preferably caused by the administration of one or more selected from the group consisting of docetaxel and capecitabine; Preferably, the hand-foot skin reaction is caused by the administration of one or more of the group consisting of EGFR inhibitors, VEGFR inhibitors, c-kit, PDGFR, ALK inhibitors, RET inhibitors and FGFR inhibitors, preferably by the administration of one or more of the group consisting of EGFR inhibitors and VEGFR inhibitors, preferably by the administration of Sora The group consisting of one or more of fenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, fruquintinib, anlotinib, erlotinib, afatinib and osimertinib.

38. Use of the compound according to any one of claims 1 to 31 or its stereoisomers, tautomers, solvates, hydrates, prodrugs, stable isotope derivatives and pharmaceutically acceptable salts, or the pharmaceutical composition according to claim 32 in the preparation of a drug for preventing and / or treating diseases related to abnormal methylation of RNA m6A; Preferably, the RNA m6A abnormal methylation-related disease is selected from skin-related diseases; Preferably, the skin-related disease is selected from diseases related to skin hyperkeratosis; Preferably, the skin hyperkeratosis-related disease is selected from skin keratin abnormal growth diseases; preferably selected from rash, itching, folliculitis, paronychia, pigmentation disorder, peeling, acne, urticaria, acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, oral ulcer and dermatitis; Preferably, the hand-foot syndrome is caused by the administration of one or more selected from the group consisting of docetaxel, capecitabine, fluorouracil, paclitaxel, cytarabine, adriamycin, oxaliplatin and doxorubicin, preferably caused by the administration of one or more selected from the group consisting of docetaxel and capecitabine; Preferably, the hand-foot skin reaction is caused by the administration of one or more of the group consisting of EGFR inhibitors, VEGFR inhibitors, c-kit, PDGFR, ALK inhibitors, RET inhibitors and FGFR inhibitors, preferably caused by the administration of one or more of the group consisting of EGFR inhibitors and VEGFR inhibitors, preferably caused by the administration of one or more of the group consisting of sorafenib, lenvatinib, axitinib, donafenib, regorafenib, apatinib, furiquintinib, anlotinib, erlotinib, afatinib and osimertinib.

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