WO2024012534A1 - Heterocyclic fused benzene ring compounds, preparation method therefor, and use thereof - Google Patents

Abstract

Provided are heterocyclic fused benzene ring compounds, a preparation method therefor, and the use thereof. The heterocyclic fused benzene ring compounds are heterocyclic compounds represented by formula I, tautomers thereof, stereoisomers thereof, hydrates thereof, solvates thereof, pharmaceutically acceptable salts thereof, or prodrugs thereof. The compounds have a good cell proliferation-inhibiting effect on cells of cutaneous squamous cell carcinoma, thus having good application prospects. (I)

Description

Heterocyclic acene compounds and their preparation methods and uses

This application claims priority from Chinese patent application 2022108285545 with a filing date of July 13, 2022. This application cites the full text of the above-mentioned Chinese patent application.

Technical field

The present invention belongs to the field of medicine. Specifically, the present invention relates to a heterocyclic acene ring compound and its preparation method and use.

Background technique

Microtubules are an important component of the cytoskeleton in eukaryotic cells and play an important role in maintaining cell morphology, signal transmission, organelle transport, cell movement, cell division and mitosis and other cellular functions.

Microtubules are composed of two types of tubulin subunits, α-tubulin and β-tubulin. α-tubulin and β-tubulin form tubulin heterodimers, which are microtubule The basic unit of assembly. Microtubule-targeting agents (MTAs) can destroy the dynamic stability and structure of microtubules, interfere with the formation of mitotic spindles, induce cell cycle arrest in the G2/M phase, and promote cell apoptosis.

Microtubules participate in many important cellular processes and have become one of the most important drug targets for the treatment of hyperproliferative diseases. Several microtubule-targeting agents approved by the US FDA, such as vinblastine and taxanes, are widely used. Treatment of multiple solid tumors and hematological malignancies, but drug resistance and dose-limiting toxicity of microtubule-targeted drugs limit their clinical efficacy. Compared with single-target drugs, dual-target inhibitors can overcome drug resistance and can improve therapeutic effects, and have become a research hotspot, such as: tubulin-SRC dual-target inhibitors, tubulin-receptor tyrosine Kinase (receptor tyrosine kinases inhibitor, RTK) dual-target inhibitor, tubulin-histone deacetylase (histone deacetylases inhibitor, HDAC) dual-target inhibitor, etc.

Actinic keratosis (AK) is a skin disease associated with prolonged exposure to ultraviolet light. AK is the second most common disease among dermatologists in the United States and is characterized by uncontrolled, mutated keratinocytes. The proliferation is considered a precancerous lesion and, if left untreated, 20% of cases may develop into cutaneous squamous cell carcinoma (SCC). Current tubulin-SRC dual-target inhibitor Tirbanibulin Clinically, local treatment of AK has significant effects and has been approved by the FDA for marketing. This indicates that the development of new tubulin-SRC dual-target inhibitors with better effects for local treatment of AK may be a promising direction.

The development of new compounds that inhibit tubulin polymerization or Src kinase is currently of great significance for the treatment of many diseases including actinic keratosis.

Contents of the invention

The object of the present invention is to provide a heterocyclic acene ring compound that is completely different from the prior art. The heterocyclic acene ring compound of the present invention has a good inhibitory effect on cell proliferation of skin squamous cancer cells, and is particularly fast in metabolism in the body and has little irritation to the skin.

In the first aspect of the present invention, there is provided a compound represented by formula I, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug:

in,

Ring B is a saturated, unsaturated or partially saturated 5-6 membered heterocyclic ring;

Y is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R ¹ ; the R ¹ is selected from halogen, hydroxyl, amino, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl and C ₁ -C ₆ alkoxy group; when R ¹ is multiple, the R ^1s are the same or different;

Z is C ₃ -C ₈ cycloalkyl, 3-11 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl, wherein the C ₃ -C ₈ cycloalkyl, 3- 11-membered heterocycloalkyl, 6-10-membered aryl, 5-10-membered heteroaryl are optionally substituted by one or more R ² ; when R ² is multiple, the R ² is the same or different;

R ² is each independently selected from halogen, hydroxyl, amino, cyano, carbonyl, oxo (=O), carboxyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl , C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkyl-OH, C ₁ -C ₆ alkyl -C(=O)-, C ₁ -C ₆ alkylcyano, C ₁ -C ₆ alkyl Oxygen, C ₁ -C ₆ haloalkoxy, C ₁ -C ₆ alkoxycarbonyl, C ₁ -C ₆ alkoxy C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy -C(=O )-C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ halocycloalkyl, C ₃ -C ₈ cycloalkylcarbonyl, C ₃ -C ₈ cycloalkyl -C ₁ -C ₆ alkyl-carbonyl, C ₃ -C ₈ cycloalkoxy, C ₃ -C ₈ halocycloalkoxy, optionally aminocarbonyl with 1 or 2 C ₁ -C ₆ alkyl groups Groups, C ₁ -C ₆ alkylsulfonyl groups, aminosulfonyl groups optionally having 1 or 2 C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkylsulfonylamino groups and any Choose an amino group with 1 or 2 C ₁ -C ₆ alkyl groups;

Q is a 6-10-membered aryl group that is unsubstituted or substituted by one or more R ³ , or a 5-10-membered heteroaryl group that is unsubstituted or substituted with one or more R ³ ;

The heteroatoms of the heteroaryl group are N, O or S, and the number of heteroatoms is 1, 2 or 3;

Each of the R ³ is independently selected from halogen, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ halocycloalkyl, C ₃ -C ₈ cycloalkoxy and C ₃ -C ₈ halocycloalkoxy; or when R ³ is multiple, substituted 6-10 membered aryl group Or a 5-10 membered heteroaryl group and R ³ together form an O-containing saturated heterocyclic ring;

R ^a , R ^b and R ^c are absent, or are each independently halogen, hydroxyl, cyano, carbonyl, oxo (=O), C ₂ -C ₆ alkenyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkyl -OH, C ₁ -C ₆ alkyl -C(=O)-, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ halocycloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy, -COO-C ₁ -C ₆ alkyl or -C(O)NR ⁴ R ⁵ ; wherein, The R ⁴ and R ⁵ are each independently hydrogen or C ₁ -C ₆ alkyl;

m, n, p, q are independently 0, 1, 2 or 3.

In the present invention, the definitions of certain substituents in the compound represented by Formula I can be as follows, and the definitions of unmentioned substituents are as described in any of the above schemes.

In a preferred embodiment of the present invention, the heteroatoms in each of the above-mentioned heterocycles, the above-mentioned heteroaryl groups and each of the above-mentioned heterocycloalkyl groups are independently selected from one of N, O and S, and the number can be 1 , 2 or 3.

Preferably, when the heteroatom contains an S atom, the lone pair of electrons on the S atom is not replaced by an O atom to form

In a preferred embodiment of the present invention, the above-mentioned O-containing saturated heterocyclic ring may be a 3-11 membered O-containing saturated heterocyclic ring.

In a preferred embodiment of the present invention, in ring B, the saturated, unsaturated or partially saturated 5-6-membered heterocyclic ring is a 5-6-membered heteroaromatic ring or a 5-6-membered heteroolefin ring. The atoms are selected from one or more of N, O and S, and the number is 1, 2 or 3.

In a preferred embodiment of the present invention, Ring B is a saturated, unsaturated or partially saturated 5-6 membered heterocyclic ring containing 1, 2 or 3 heteroatoms;

Preferably, the heteroatoms are selected from heteroatoms of N, O and S; when there are multiple heteroatoms, the heteroatoms are the same or different;

Preferably, Ring B is dihydrofuran (e.g. Indicates the formation of a paracyclic ring with a benzene ring), furan (for example Indicates the formation of a paracyclic ring with the benzene ring here), dihydrothiophene (such as Indicates the formation of a paracyclic ring with the benzene ring here), thiophene (e.g. means forming a parallel ring with a benzene ring), pyrrole, dihydropyrrole, imidazole, dihydrimidazole, oxazole, dihydroxazole, thiazole, dihydrothiazole, piperidine, tetrahydropyridine, morpholine, piperazine , hexahydropyrimidine, tetrahydropyrimidine, triazine, tetrahydrotriazine, tetrahydropyran, dihydropyran, tetrahydrothiopyran or dihydrothiopyran;

More preferably, Ring B is dihydrofuran, furan, dihydrothiophene or thiophene.

In a preferred embodiment of the present invention, in _Y , the C 1 -C 6 alkyl group in the C _{1 -C 6} alkyl group that is unsubstituted or substituted by one or more R ¹ is methyl, ethyl, n- Propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

In a preferred embodiment of the present invention, Y is an unsubstituted or methyl-substituted C ₁ -C ₆ alkyl group;

Preferably, Y is methyl.

In a preferred embodiment of the present invention, Z is a 3-11 membered saturated heterocyclic ring that is unsubstituted or substituted by R ² ;

The R ² is C ₁ -C ₆ alkyl or C ₃ -C ₈ cycloalkyl-C ₁ -C ₆ alkyl-carbonyl, and the substitution is 1 or 2; when the substituent is 2, The substituents are the same or different;

Preferably, the 3-11 membered saturated heterocyclic ring has 1 to 3 heteroatoms selected from N, O and S.

In a preferred embodiment of the present invention, Z is a 5-10 membered saturated heterocyclic ring that is unsubstituted or substituted by R ² ;

The 5-10 membered saturated heterocyclic ring is a single ring, a paracyclic ring, a spiro ring or a bridged ring.

In a preferred embodiment of the present invention, Z is a 5-8-membered monocyclic ring, a 6-10-membered spirocyclic ring, a 6-10-membered bridged ring or an 8-10-membered paracyclic ring that is unsubstituted or substituted by ^R2 .

In a preferred embodiment of the present invention, in Z, the 3-11-membered heterocycloalkyl group is a 5, 6, 7 or 8-membered saturated monocyclic heterocyclic ring, and the 6-, 7-, 8-, 9- or 10-membered heterocycloalkyl group is Saturated spiro or bridged heterocycle, or 8-, 9- or 10-membered saturated pendant heterocycle, having 1 to 3 A heteroatom selected from N, O and S, for example, a 6-, 7- or 8-membered saturated monocyclic heterocycle.

Preferably, the 5, 6, 7 or 8-membered saturated monocyclic heterocyclic ring is morpholinyl, 1,4-oxaheptanyl, azetidinyl, pyrrolidinyl or piperazinyl , such as morpholinyl.

In a preferred embodiment of the present invention, in R ² , the halogen is fluorine, chlorine or bromine, such as fluorine.

In a preferred embodiment of the present invention, in R ² , _C 1 -C 6 in the C ₁ -C ₆ alkyl group and the C _{3 -C 8} cycloalkyl-C ₁ -C ₆ alkyl-carbonyl group Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

In a preferred embodiment of the present invention, in R ² , the C ₁ -C ₆ alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy group, sec-butoxy or tert-butoxy, such as methoxy.

In a preferred embodiment of the present invention, in R ² , the C ₃ -C ₈ cycloalkyl group in the C 3 _-C 8 cycloalkyl _{- C 1} -C ₆ alkyl-carbonyl group is cyclopropanyl, cyclobutyl Alkyl, cyclopentyl or cyclohexyl, for example cyclopropyl.

Preferably, R ² is each independently selected from halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy and C ₃ -C ₈ cycloalkyl-C ₁ -C ₆ alkyl-carbonyl.

Preferably, R ² is each independently selected from fluorine, methyl, methoxy and The number is 1 or 2.

In a preferred embodiment of the present invention, Z is

Preferably, Z is

In a preferred embodiment of the present invention, in Q, the 6-10-membered aryl group in the 6-10-membered aryl group that is unsubstituted or substituted by one or more R ³ is phenyl or naphthyl, such as phenyl base.

In a preferred embodiment of the present invention, in Q, the 5-10-membered heteroaryl group that is unsubstituted or substituted by one or more R ³ is a 5-6-membered heteroaryl group, and the 5-6-membered heteroaryl group The aryl heteroatom is one or more of N, O or S, and the number of heteroatoms is 1, 2 or 3.

In a preferred embodiment of the present invention, in R ³ , the halogen is fluorine, chlorine or bromine, such as fluorine.

In a preferred embodiment of the present invention, Q is a phenyl group that is unsubstituted or substituted by R ³ , a 5-6-membered heteroaryl group that is unsubstituted or substituted by R ³ ;

The heteroatoms of the heteroaryl group are N, O or S, and the number of heteroatoms is 1, 2 or 3;

The R ³ substitution is one or more substitutions, and the R ³ is each independently selected from hydrogen, halogen, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy; or, when When there are multiple R ^3s , the substituted phenyl group or 5-6-membered heteroaryl group and R ³ together form an O-containing saturated heterocyclic ring;

Preferably, Q is unsubstituted or phenyl substituted by R ³ ;

Preferably, Q is unsubstituted or fluorine-substituted phenyl.

In a preferred embodiment of the invention, Q is

In a preferred embodiment of the present invention, in R ^a , the halogen is fluorine, chlorine or bromine, such as fluorine or chlorine.

In a preferred embodiment of the present invention, in R ^a , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert. Butyl, such as methyl.

In a preferred embodiment of the present invention, in R ^a , the C ₂ -C ₆ alkynyl group is ethynyl, propynyl or butynyl, for example

In a preferred embodiment of the invention, R ^a does not exist, or is independently halogen, hydroxyl, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl, C ₂ -C ₆ alkyne base or C ₁ -C ₆ haloalkyl;

Preferably, R ^a does not exist, or is independently halogen, cyano, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkynyl;

Preferably, R ^a does not exist, or is independently fluorine, chlorine, cyano, methyl or propynyl.

In a preferred embodiment of the present invention, R ^b does not exist, or is independently halogen, hydroxyl, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl, C ₂ -C ₆ alkyne base or C ₁ -C ₆ haloalkyl;

Preferably, R ^b does not exist, or is independently halogen, cyano or C ₁ -C ₆ alkyl;

Preferably, R ^b does not exist or is fluorine.

In a preferred embodiment of the present invention, in R ^c , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert. Butyl, such as methyl.

In a preferred embodiment of the present invention, R ^c is absent, or is independently halogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl, C ₁ -C ₆ haloalkyl or C ₁ -C ₆ alkoxy;

Preferably, R ^c does not exist, or is independently halogen or C ₁ -C ₆ alkyl;

Preferably, R ^c is absent or is methyl.

In a preferred embodiment of the invention, for

In a preferred embodiment of the invention,

In a preferred embodiment of the present invention, the compound represented by formula I has the structure represented by formula I-1:

where the dotted lines represent optional double bonds ( be a double bond or a single bond);

When the dashed line does not represent a double bond (i.e. is a single bond), X ₁ and X ₂ are independently C, NH, O or S; at least one of X ₁ and X ₂ is not C;

When the dotted line represents a double bond, X ₁ is C and X ₂ is NH, O or S;

Ra, ^Rb , ^Rc , ^Y , Z, Q, m, n, p and q are as defined in the first aspect of the invention.

Preferably, X ₁ is C.

More preferably, X ₁ is C, and X ₂ is O or S.

In a preferred embodiment of the present invention, the compound represented by formula I has the structure:

Among them, X ₁ and X ₂ are independently C, NH, O or S; at least one of X ₁ and X ₂ is not C;

Ra, ^Rb , ^Rc , ^Y , Z, Q, m, n, p and q are as defined in the first aspect of the invention.

In a preferred embodiment of the present invention, the compound represented by formula I has the structure represented by formula II-1:

Preferably, it has the structure:

Wherein, Ra, ^Rb , ^Rc , Y, Z, Q, m, ⁿ , p and q are defined as described in the first aspect of the present invention.

More preferably, in the formula II-1, formula II-1a and formula II-1b, R ^c is absent or C ₁ -C ₆ alkyl; Z is morpholinyl; Q is phenyl.

optimally, for

R ^b does not exist.

In a preferred embodiment of the present invention, the compound represented by formula I has the structure represented by formula II-2:

Wherein, Ra, ^Rb , ^Rc , Y, Z, Q, m, ⁿ , p and q are defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the compound represented by formula I has the structure represented by formula II-3:

Wherein, Ra, ^Rb , ^Rc , Y, Z, Q, m, ⁿ , p and q are defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the formula II-1 has the structure shown in formula II-1c:

In formula II-1c, n is 0 or 1; p is 0 or 1; Ra and Rc are each independently methyl, fluorine, chlorine or cyano.

Preferably, the formula II-1c has the structure shown in formula II-1c-1 or II-1c-2

Wherein, n is 0 or 1; p is 0 or 1; Ra and Rc are each independently methyl, fluorine, chlorine or cyano.

In a preferred embodiment of the present invention, the compound represented by formula I is selected from any of the following compounds:

In a preferred embodiment of the present invention, the compound represented by formula I is: The compound that peaks first in chiral separation or chiral analysis;

or, Compounds that elute late in chiral separations or chiral analyses.

The chiral separation and chiral analysis conditions are particularly preferably the following detection parameters:

Detection parameter 1: Chromatographic column: Chiralcel OD; mobile phase A: CO ₂ , mobile phase B: MeOH and acetonitrile containing 0.05% diethylamine (for example, the volume ratio of MeOH to acetonitrile is 2:1);

More preferably, the separation gradient is supercritical fluid carbon dioxide of a mixed solution of 40% MeOH and acetonitrile containing 0.05% diethylamine; flow rate: 3mL/min; detector: PDAColumn; column temperature: 35°C; column pressure: 100Bar .

Detection parameter 2: Chromatographic column: Chiralcel OD-3 50×4.6mm I.D., 3μm; mobile phase A: supercritical fluid carbon dioxide, mobile phase B: ethanol containing 0.05% diethylamine; retention of the compound that peaks first The time is 1.204min; the retention time of the compound that peaks later is 2.010min;

More preferably, gradient elution: supercritical fluid carbon dioxide containing 0.05% diethylamine in ethanol, with a ratio from 5% to 40%; flow rate: 3mL/min; detector: PDAColumn; column temperature: 35°C; column Pressure: 100Bar;

The % in the above detection conditions are all volume percentages.

Those skilled in the art can correspondingly select and adjust chiral analysis or separation conditions based on the above detection parameters to obtain the above-mentioned compounds that peak first and the above-mentioned compounds that peak later. For example, when preparing by high-performance liquid phase method, they can be The high-performance liquid phase separation conditions among the above-mentioned detection parameters can also be used for other separation conditions, as long as the detection parameters of the obtained products are consistent with the above-mentioned detection parameters.

In a preferred embodiment of the present invention, the compound represented by formula I is any of the following compounds:

A second aspect of the present invention provides a pharmaceutical composition, characterized in that the pharmaceutical composition includes: the compound represented by formula I described in the first aspect of the present invention, its tautomers, and its stereoisomers. body, its hydrates, its solvates, its pharmaceutically acceptable salt or prodrug thereof; and a pharmaceutically acceptable carrier.

The third aspect of the present invention provides the compound represented by formula I as described in the first aspect of the present invention, its tautomers, its stereoisomers, its hydrates, its solvates, and its pharmaceutically acceptable The use of salts or prodrugs thereof, or the use of the pharmaceutical composition according to the second aspect of the present invention, the use includes: inhibiting tubulin polymerization or Src kinase; and/or preventing and/or treating tubulin Polymerization-mediated or Src kinase-related diseases; and/or, preparation of drugs or drugs for inhibiting tubulin polymerization or Src kinase, and/or preventing and/or treating tubulin polymerization-mediated or Src kinase-related diseases. Composition or preparation.

Preferably, the tubulin polymerization-mediated or Src kinase-related diseases include: one, two or more of tumors, skin diseases and/or other diseases.

Preferably, the tumors include but are not limited to: solid tumors, sarcomas, hematological cancers, subtypes include breast cancer, ovarian cancer, prostate cancer, cervical cancer, testicular cancer, colon cancer, colorectal cancer, liver cancer, non- Small cell lung cancer, squamous cell carcinoma, small cell lung cancer, gastric cancer, gastrointestinal stromal tumor, pancreatic cancer, bladder cancer, germ cell tumor, mast cell tumor, mastocytosis, glioblastoma, neuroblastoma tumour, astrocytoma, melanoma, B-cell lymphoma, T-cell lymphoma, slowly progressive lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, Chronic myeloid leukemia, chronic lymphocytic leukemia, myeloma, and/or myelodysplastic syndrome

Preferably, the skin diseases include but are not limited to: actinic keratosis, psoriasis, atopic dermatitis, psoriasis, vitiligo, roseola and/or systemic lupus erythematosus.

Preferably, the other diseases include but are not limited to: autoimmune diabetes, diabetic retinopathy, liver fibrosis, pulmonary fibrosis (including idiopathic pulmonary fibrosis, etc.), renal fibrosis, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinocerebellar degeneration, atherosclerosis, anemia, sickle cell anemia, thalassemia, osteoarthritis, rheumatoid arthritis, malaria, contusion Anthroplasmosis, helminthiasis, protozoal infections, multiple sclerosis, lupus, asthma, allergic rhinitis and/or inflammatory bowel disease.

In a fourth aspect of the present invention, there is provided a method for inhibiting tubulin polymerization or Src kinase, or preventing and/or treating microtubule oligomerization or Src kinase-related diseases, including the step of: administering the first aspect of the present invention to a subject in need. In one aspect, the compound represented by formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug.

In the fifth aspect of the present invention, there is provided a compound represented by formula I as described in the first aspect of the present invention, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutical The use of acceptable salts or prodrugs thereof, or the use of the pharmaceutical composition according to the second aspect of the present invention, for the preparation of medicaments for the treatment and/or prevention of actinic keratosis.

The actinic keratosis is preferably squamous skin cancer.

In the sixth aspect of the present invention, there is provided a method for treating and/or preventing the above-mentioned anti-actinic keratosis, which includes administering a compound of formula I as described in the first aspect of the present invention to a subject in need. , its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or the use of its prodrug, or the pharmaceutical composition described in the second aspect of the present invention.

In the seventh aspect of the present invention, there is provided a compound represented by the above formula I, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug. Preparation method, which includes the following steps:

In the presence of a basic reagent (such as sodium carbonate) and a palladium catalyst (such as tetrakis(triphenylphosphine)palladium), compound II and compound II Carry out Suzuki reaction in a solvent to obtain the compound represented by formula I;

_Where ^{, _ _}

In an eighth aspect of the present invention, a compound represented by formula II or a compound represented by formula IV is provided:

Wherein, X ₄ is halogen (such as chlorine, bromine, iodine), and ring B, ^Ra , R ^c , n, p and q are as defined above.

The compound represented by formula II is preferably

The compound represented by formula IV is preferably

Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.

Terms and definitions

Unless otherwise stated, the definitions of groups and terms recorded in the specification and claims of this application include their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples. etc., can be arbitrarily combined and combined with each other. Such combinations and combined group definitions and compound structures should fall within the scope described in the specification of this application.

Unless otherwise defined, all technical and scientific terms herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. Unless otherwise indicated, all patents, patent applications, and published materials cited in their entirety are hereby incorporated by reference in their entirety. If there are multiple definitions for a term herein, the definitions in this chapter shall prevail.

It should be understood that the foregoing brief description and the following detailed description are exemplary and are for explanation only, and do not limit the subject matter of the present invention in any way. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, unless otherwise clearly stated in the text, As used in the specification and claims, the singular forms include plural referents. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the use of the term "includes" and other forms such as "includes,""contains," and "contains" is not limiting.

Unless otherwise stated, conventional methods within the skill in the art, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy and pharmacological methods, were used. Unless specific definitions are given, the terminology employed herein in connection with the description of analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry is known in the art. Standard techniques may be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and treatment of patients. For example, the manufacturer's instructions for using the kit can be used, or the reaction and purification can be carried out in accordance with methods known in the art or the instructions of the present invention. The above techniques and methods can generally be implemented in accordance with conventional methods well known in the art, as described in the various general and more specific documents cited and discussed in this specification. In this specification, groups and their substituents may be selected by those skilled in the art to provide stable moieties and compounds.

When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes substituents that are chemically equivalent when the structural formula is written from right to left. For example, CH ₂ O is equivalent to OCH ₂ . As used in this article, Indicates the attachment site of the group.

The section headings used in this article are for the purpose of organizing the article only and should not be construed as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and theses, are hereby incorporated by reference in their entirety.

In addition to the foregoing, when used in the description and claims of this application, the following terms have the meanings shown below unless otherwise specified.

When the numerical range described in the specification and claims of this application is understood as an "integer," it should be understood that the two endpoints of the range and each integer within the range are recorded. For example, "integers from 1 to 6" should be understood as describing each integer of 0, 1, 2, 3, 4, 5, and 6. When a numerical range is understood to mean a "number," it should be understood that both endpoints of the range are recited as well as every integer within the range and every decimal within the range. For example, "numbers from 1 to 10" should be understood as not only recording every integer from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, but also recording at least each of the integers corresponding to The sum of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9.

In this application, "substitution" means that a hydrogen atom in a molecule is replaced by a different atom or group, or that a lone pair of electrons of an atom in a molecule is replaced by another atom or group. For example, the lone pair of electrons on the S atom can be replaced by an O atom to form

In this application, "saturated, partially saturated or unsaturated" includes substituents that are saturated with hydrogen, substituents that are completely unsaturated with hydrogen, and substituents that are partially saturated with hydrogen.

In this application, the term "halogen" means fluorine, chlorine, bromine, iodine, alone or as part of another substituent.

As used herein, the term "cyano", alone or as part of another substituent, means -CN.

As used herein, the term "amino", alone or as part of another substituent, means _-NH2 .

As used herein, the term "alkyl", alone or as part of another substituent, means consisting solely of carbon atoms and hydrogen atoms, A straight or branched hydrocarbon chain group containing no unsaturated bonds, having, for example, 1 to 6 carbon atoms and being connected to the rest of the molecule by a single bond. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl and hexyl. Alkyl groups may be unsubstituted or substituted with one or more suitable substituents. The alkyl group may also be an isotopomer of the naturally abundant alkyl group that is rich in isotopes of carbon and/or hydrogen (ie, deuterium or tritium). As used herein, the term "alkenyl" refers to an unbranched or branched monovalent hydrocarbon chain containing one or more carbon-carbon double bonds. As used herein, the term "alkynyl" refers to an unbranched or branched monovalent hydrocarbon chain containing one or more carbon-carbon triple bonds.

The term "C ₁ -C ₆ alkyl" when used alone or as part of other substituents is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-Methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl base, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. or their isomers. In particular, said groups have 1, 2 or 3 carbon atoms ("C ₁ -C ₃ alkyl"), for example methyl, ethyl, n-propyl or isopropyl.

"Haloalkyl," when used alone or as part of other substituents, refers to branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms substituted with one or more halogens (e.g., -CvFw, where v=1 to 3, w=1 to (2v+1)). Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2,2,2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl.

The term "cycloalkyl" or "carbocyclyl" when used alone or as part of another substituent refers to a cyclic alkyl group. The term "mn-membered cycloalkyl" or "C _m -C _n cycloalkyl" is understood to mean a saturated, unsaturated or partially saturated carbocyclic ring having m to n atoms. For example, "3-15 membered cycloalkyl" or "C ₃ -C ₁₅ cycloalkyl" refers to a cyclic alkyl group containing 3 to 15, 3 to 9, 3 to 6 or 3 to 5 carbon atoms, which May contain 1 to 4 rings. "5-8 membered cycloalkyl" contains 5-8 carbon atoms. Including single ring, two rings, three rings, spiro rings or bridged rings. Examples of unsubstituted cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and adamantyl, or bicyclic hydrocarbon groups such as decalin rings. Cycloalkyl groups may be substituted by one or more substituents. In some embodiments, a cycloalkyl group can be a cycloalkyl group fused to an aryl or heteroaryl group. The term "C ₃ -C ₆ cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms, including fused or bridged polycyclic systems. For example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

The term "heterocycloalkyl" when used alone or as part of other substituents refers to a cycloalkyl group in which one or more (in some embodiments 1 to 3) carbon atoms are substituted with a heteroatom. Atoms such as, but not limited to, N, O, S, and P. The term "mn-membered heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms, wherein the heterocyclic atoms are selected from N, O, S, P, preferably from N, O or S. For example, the term "4-8 membered heterocycloalkyl" or "C ₄ -C ₈ heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having 4 to 8 atoms, where 1, 2 , 3, or 4 ring atoms are selected from N, O, S, P, preferably selected from N, O or S. "4-10 membered heterocyclyl" means a saturated, unsaturated or partially saturated ring with 4 to 10 atoms. In some embodiments, the heterocycloalkyl group may be a heterocycloalkyl group fused to an aromatic or heteroaromatic group. When a prefix such as 4-8 membered or 4-10 membered is used to indicate heterocycloalkyl, the number of carbons is also meant to include heteroatoms. Including single ring, two rings, three rings, spiro rings or bridged rings. Examples of heterocycloalkyl groups are: pyrrolidinyl, tetrahydrofuryl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrrolyl, azetidinyl, thiazolidinyl, oxazolidinyl , piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, azepanyl, diazepanyl, Oxazepanyl, etc. The term "heterocycloalkyl" may be used interchangeably with the terms "heterocycloalkyl", "heterocycle" and "heterocyclocycloalkane".

The term "partially saturated heterocycle" when used alone or as part of other substituents refers to a monocyclic group having heteroatoms (the monocyclic group has a double bond but is not aromatic), preferably containing 1, A monocyclic ring with 2 or 3 ring heteroatoms independently selected from N, O and S. Examples of partially saturated heterocycles are: dihydrofuryl, dihydrothienyl, dihydropyrrolyl, dioxolyl, dihydroimidazolyl, dihydropyrazolyl, dihydrothiazolyl, dihydrogen Isothiazolyl, dihydroxadiazolyl, dihydrothiazolyl, dihydrotriazolyl, dihydrotetrazolyl, tetrahydropyridinyl, 3,4-dihydro-2H-pyran, pyranyl , Thiopyranyl, dihydropyridyl, dihydropyrazinyl, dihydropyrimidinyl, oxazinyl, dihydrotetrazolyl, etc.

The term "alkenyl" when used alone or as part of other substituents refers to a straight or branched chain monovalent hydrocarbon radical of two to forty carbon atoms having at least one carbon-carbon sp2 double bond (e.g., C ₂ - C ₆ alkenyl, another example is C ₂ -C ₄ alkenyl), and includes groups having "cis" and "trans" orientations or "E" and "Z" orientations. Examples of alkenyl groups include, but are not limited to, vinyl and allyl.

The term "alkynyl" when used alone or as part of other substituents refers to a straight or branched chain monovalent hydrocarbon radical of two to forty carbon atoms (e.g., C ₂ -C ₆ ) having at least one carbon-carbon sp triple bond. Alkynyl, another example is C ₂ -C ₄ alkynyl). Examples of alkynyl groups include, but are not limited to, ethynyl and propynyl.

The term "alkoxy" when used alone or as part of another substituent refers to the group ^-ORx , where ^Rx is "alkyl" as defined above.

The term "oxo" when used alone or as part of other substituents means that the two hydrogens on the methylene group are replaced by oxygen, that is, the methylene group is replaced by a carbonyl group.

The term "aryl" when used alone or as part of other substituents, refers to a monocyclic or polycyclic carbocyclic ring having from 6 to 20 carbon atoms, at least one of which is aromatic. When one of the rings is non-aromatic, the group can be attached through the aromatic ring or through the non-aromatic ring. Examples of aryl groups include, but are not limited to: phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthrenyl, anthracenyl, and acenaphthyl.

The term "heteroaryl ring" when used alone or as part of other substituents refers to a monocyclic or polycyclic carbocyclic ring in which at least one ring atom is a heteroatom independently selected from oxygen, sulfur and nitrogen and the remaining ring atoms are C, at least one of the rings is aromatic. The group may be a carbon group or a heteroatom group (ie it may be C-linked or N-linked, wherever possible). When one of the rings is non-aromatic, the group can be attached through the aromatic ring or through the non-aromatic ring. Examples of heteroaryl groups include, but are not limited to: imidazolyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furyl, thienyl, Benzothienyl, benzofuryl, quinolyl, isoquinolyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, N -Methyl pyrrolyl and tetrahydroquinoline. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic," "heteroaryl" or "heteroaryl."

The term "cyclo-and" when used alone or as part of other substituents, refers to fused bicyclic compounds in which the two rings share two adjacent atoms. In other words, the rings share a covalent bond, i.e. the bridgehead atoms are directly connected (e.g. α-thujane and decalin). Examples of parallel loops include but are not limited to:

The term "spiro", when used alone or as part of other substituents, refers to a polycyclic group in which the single rings share one carbon atom (called a spiro atom) and may contain one or more double bonds, but no single ring has A fully conjugated π electron system. According to the number of shared spiro atoms between the rings, the spirocycloalkyl group is divided into a single spirocycloalkyl group, a double spirocycloalkyl group or a polyspirocycloalkyl group, and is preferably a single spirocycloalkyl group and a double spirocycloalkyl group. Non-limiting examples of spirocycloalkyl groups include:

Also included are spirocycloalkyl groups in which a single spirocycloalkyl group and a heterocycloalkyl group share a spiro atom. Non-limiting examples include:

The term "bridged ring" refers to a cyclic hydrocarbon in which any two rings in a compound share two carbon atoms that are not directly connected. According to the number of constituent rings, it is divided into bicyclic hydrocarbons, tricyclic hydrocarbons, tetracyclic hydrocarbons, etc. Non-limiting examples include:

"Haloalkyl," when used alone or as part of other substituents, refers to branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms substituted with one or more halogens (e.g., -CvFw, where v=1 to 3, w=1 to (2v+1)). Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2,2,2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl.

The term "inert solvent" includes, but is not limited to: toluene, benzene, water, methanol, ethanol, isopropyl alcohol, ethylene glycol, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran dichloromethane, chloroform, 1 , 2-dichloroethane, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, dioxane, or combinations thereof.

The compounds provided herein include intermediates useful in preparing the compounds provided herein, which contain reactive functional groups (such as, but not limited to, carboxyl, hydroxyl and amino moieties), and also include protected derivatives thereof. "Protected derivatives" are those compounds in which one or more reactive sites are blocked by one or more protecting groups (also called protecting groups). Suitable protecting groups for the carboxyl moiety include benzyl base, tert-butyl, etc., as well as isotopes, etc. Suitable amino and amido protecting groups include acetyl, trifluoroacetyl, tert-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable hydroxyl protecting groups include benzyl and the like. Other suitable protecting groups are known to those of ordinary skill in the art.

As used herein, "optional" or "optionally" means that the subsequently described event or condition may or may not occur, and that the description includes both instances in which the event or condition does and does not occur. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl groups.

In this application, the term "salt" or "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue without multiple toxicity, irritation, allergic reactions, or other problems or complications, commensurate with a reasonable benefit/risk ratio.

"Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that can maintain the biological effectiveness of the free acid without other side effects. In addition to pharmaceutically acceptable salts, other salts are contemplated by the present invention. They may serve as intermediates in the purification of the compounds or in the preparation of other pharmaceutically acceptable salts or may be used in the identification, characterization or purification of the compounds of the invention.

The term "amine salt" refers to the product obtained by neutralizing an alkyl primary, secondary or tertiary amine with an acid. The acid includes an inorganic acid or an organic acid as described in this application.

The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereoisomers and conformational isomers.

Depending on the choice of starting materials and methods, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, for example as pure optical isomers, or as a mixture of isomers, for example as racemic and non-racemic isomers. A mixture of enantiomers, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or centers) in the molecule. The prefixes D and L or (+) and (–) are symbols used to designate the rotation of plane-polarized light caused by a compound, where (–) or L indicates that the compound is levorotatory. Compounds prefixed with (+) or D are dextrorotatory.

When the bond to a chiral carbon in the formula of the present invention is drawn as a straight line, it should be understood that the two configurations of the chiral carbon (R) and (S) and the enantiomerically pure compounds thereof and Both mixtures are included within the scope of this general formula. The schematic representation of racemic or enantiopure compounds herein is taken from Maehr, J. Chem. Ed. 1985, 62: 114-120. Use wedge-shaped bonds and dashed-line bonds to represent the absolute configuration of a stereocenter.

The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible species. Proton transfer tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually yield a mixture whose physical and chemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form is dominant; in phenols, the enol form is dominant. The present invention encompasses all tautomeric forms of the compounds.

As used herein, "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for delivering a biologically active compound to a mammal, such as a human. The medium includes a pharmaceutically acceptable carrier. The purpose of pharmaceutical compositions is to facilitate administration to organisms and facilitate the absorption of active ingredients to exert biological activity.

In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener approved by the relevant government regulatory authorities as acceptable for human or livestock use , diluents, preservatives, dyes/colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents or emulsifiers.

The term "solvate" means that the compound of the present invention or its salt includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces, and when the solvent is water, it is a hydrate.

The term "prodrug" refers to a compound of the invention that can be converted to a biologically active compound under physiological conditions or by solvolysis. The prodrugs of the present invention are prepared by modifying the functional groups in the compound, and the modifications can be removed by conventional procedures or in vivo to obtain the parent compound. Prodrugs include compounds in which a hydroxyl group or amino group in the compound of the present invention is connected to any group. When the prodrug of the compound of the present invention is administered to a mammalian individual, the prodrug is cleaved to form free hydroxyl groups and free hydroxyl groups, respectively. of amino.

The compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, compounds can be labeled with radioactive isotopes such as deuterium ( ² H), tritium ( ³ H), iodine-125 ( ¹²⁵ I) or C-14 ( ¹⁴ C). All variations in the isotopic composition of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.

The term "excipient" means a pharmaceutically acceptable inert ingredient. Examples of types of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients can enhance the handling properties of pharmaceutical formulations, i.e. make the formulation more suitable for direct compression by increasing flowability and/or viscosity.

As used herein, the term "treatment" and other similar synonyms include the following meanings:

(i) prevent the occurrence of a disease or condition in mammals, particularly where such mammals are susceptible to but have not been diagnosed as suffering from the disease or condition;

(ii) To inhibit a disease or condition, that is, to arrest its progression;

(iii) Alleviation of a disease or condition, i.e., resolution of the disease or condition; or

(iv) Reduce the symptoms of the disease or condition.

The term "prevention" refers to the reduction of the risk of acquiring or developing a disease or disorder.

The term "patient" refers to any animal, preferably a mammal, to which a compound or composition is or has been administered in accordance with embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being preferred.

The term "therapeutically effective amount" refers to an amount of a compound that, when administered to a patient, is sufficient to effectively treat the disease or condition described herein. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, and can be adjusted as necessary by one skilled in the art.

In the reaction of each step, the reaction temperature can be appropriately selected based on the solvent, starting materials, reagents, etc., and the reaction time can also be appropriately selected based on the reaction temperature, solvent, starting materials, reagents, etc. After the reaction of each step is completed, the target compound can be separated and purified from the reaction system according to common methods, such as filtration, extraction, recrystallization, washing, silica gel column chromatography and other methods. Without affecting the next step of the reaction, the target compound can also directly enter the next step of the reaction without separation and purification.

On the basis of not violating common sense in the field, the above preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

The reagents and raw materials used in the present invention are all commercially available.

The positive progressive effect of the present invention is that: after extensive and in-depth research, the inventor unexpectedly developed a compound with a completely new structure or a pharmaceutically acceptable salt thereof, as well as preparation methods and uses;

Through cell proliferation inhibition test, the compound of the present invention can significantly inhibit the proliferation of human skin squamous cells; further through mouse pharmacokinetics and skin irritation test, the compound of the present invention shows pharmacokinetic properties consistent with topical administration to the skin. , rapid metabolism in the body, low skin irritation reaction, low potential toxicity, and good medicinal properties;

The method of the compound shown in I, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug of the present invention is simple to operate, has high yield and purity. High, it can be used for industrial production of medicine.

Description of drawings

Figure 1 shows the results of the skin irritation test of the compound.

Detailed ways

The present invention is further described below by means of examples, but the present invention is not limited to the scope of the described examples. Experimental methods that do not indicate specific conditions in the following examples should be selected according to conventional methods and conditions, or according to product specifications.

Symbol or unit:

IC ₅₀ : half inhibitory concentration, refers to the concentration that reaches half of the maximum inhibitory effect

M: mol/L, for example, n-butyllithium (14.56mL, 29.1mmol, 2.5M n-hexane solution) represents a n-hexane solution of n-butyllithium with a molar concentration of 2.5mol/L.

N: Normal concentration, for example, 2N hydrochloric acid represents 2mol/L hydrochloric acid solution

Reagents:

MeOH: methanol

ACN: Acetonitrile

DEA: Diethylamine

Intermediate A1: Preparation of N-benzyl-2-(5-bromopyridin-2-yl)acetamide (A1)

The synthetic route of intermediate A1 is as follows:

Step 1: Preparation of 2-(5-bromopyridin-2-yl)acetonitrile

Add the raw material acetonitrile (19.6g, 477mmol) to anhydrous tetrahydrofuran (500mL) at room temperature, cool to -78°C, slowly add n-butyllithium (170mL, 426mmol, 2.5M) dropwise, and the reaction solution is at -78°C. Stir for 1 hour, and then dropwise add the raw material 5-bromo-2-fluoropyridine (30g, 170mmol) in tetrahydrofuran (100mL). After the dropwise addition is completed, slowly raise the temperature to room temperature, and continue stirring for 2h. After the spot plate detection reaction is completed, pour the reaction solution into saturated ammonium chloride solution/ethyl acetate (300mL/300mL), separate the liquids, extract the aqueous phase with ethyl acetate (200mL×3), combine the organic phases, and The phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by column chromatography to obtain 2-(5-bromopyridin-2-yl)acetonitrile (22.5 g, yield 67%).

Step 2: Preparation of methyl 2-(5-bromopyridin-2-yl)acetate

The raw material 2-(5-bromopyridin-2-yl)acetonitrile (30g, 152mmol) was added to anhydrous methanol (200mL) at room temperature, concentrated sulfuric acid (25mL, 457mmol) was slowly added dropwise, and then heated to reflux and stirred for 24h. After the spot plate monitoring reaction is completed, the reaction solution is concentrated under reduced pressure to remove methanol. The residue is dissolved in dichloromethane (500 mL). The organic phase is sequentially mixed with water (200 mL), saturated sodium bicarbonate solution (200 mL) and saturated brine (200 mL). Wash, and the organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated. The residue is separated and purified by column chromatography to obtain methyl 2-(5-bromopyridin-2-yl)acetate (31 g, yield 88%).

Step 3: Preparation of N-benzyl-2-(5-bromopyridin-2-yl)acetamide

Benzylamine (39.1g, 365mmol) was added to methyl 2-(5-bromopyridin-2-yl)acetate (28g, 122mmol) at room temperature, then heated to 125°C and stirred for 15h. After the reaction was completed, the plate was detected. The reaction solution was cooled to room temperature, and ethyl acetate/petroleum ether (100mL/100mL) was slowly added under stirring. The precipitated solid was filtered to obtain N-benzyl-2-(5-bromopyridin-2-yl)acetamide ( 28.5g, yield 77%).

¹ H NMR (400MHz, DMSO-d6): δ8.66–8.53(m,2H),7.97(dd,J＝8.3,2.5Hz,1H),7.38–7.19(m,6H),4.26(d,J ＝5.9Hz,2H),3.66(s,2H)

LC-MS,M/Z(ESI):306.9[M+H] ⁺

Example 1: N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)acetamide (I-1 )

The synthetic route of compound I-1 is as follows:

Step 1: Synthesis of 5-bromo-2-(bromomethyl)-2,3-dihydrobenzofuran

A single-mouth reaction bottle, wrapped with aluminum foil, add 2-allylphenol (3.4g, 25mmol), dichloromethane (250mL), trifluoroacetic acid (12.5mL) and 1,8-diazobispiro[ 5.4.0] Undec-7-ene (3.8 mL, 2.5 mmol), then N-bromosuccinimide (14.7 g, 82.5 mmol) was added portionwise. The reaction solution was stirred at 0°C for 4 hours. The reaction solution was diluted by adding 200 mL of methylene chloride, and washed with NaHSO ₃ aqueous solution (2 mol/L, 2*200 mL) and water (100 mL) in sequence. The organic phase was dried over _MgSO4 , filtered and concentrated. The crude product was separated and purified using a silica gel column (petroleum ether: ethyl acetate (V/V) = 95:5) to obtain 5-bromo-2-(bromomethyl)-2,3-dihydrobenzofuran (2.1g, Yield 30%).

Step 2: Synthesis of 4-((5-bromo-2,3-dihydrobenzofuran-2-yl)methyl)morpholine

Add the raw materials 5-bromo-2-(bromomethyl)-2,3-dihydrobenzofuran (0.6g, 2.1mmol) and potassium carbonate (0.57g, 4.2mmol) to anhydrous acetonitrile (15mL) at room temperature. in, then add morpholine (0.22g, 2.47mmol), heat to 80°C and stir for 12 hours. After the reaction is completed, concentrate to remove the solvent, and the crude product is purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) = 95:5), obtaining 4-((5-bromo-2,3-dihydrobenzofuran-2-yl)methyl)morpholine (0.5 g, yield 81%).

Step 3: 4-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzo Synthesis of Furan-2-yl)methyl)morpholine

The raw materials 4-((5-bromo-2,3-dihydrobenzofuran-2-yl)methyl)morpholine (0.5g, 1.68mmol) and pinacol diborate (0.88g, 3.47 mmol) was added to 1,4-dioxane (5mL), then potassium acetate (0.34g, 3.47mmol) was added, and 1,1'-bis(diphenylphosphino)ferrocene dichloride was added under nitrogen protection. Palladium (II) (42 mg, 58 μmol) was added, heated to 100°C, and stirred for 6 hours. The reaction solution was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (50 mL Plate separation and purification (petroleum ether: ethyl acetate (V/V) = 5:1) gave 4-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane) Cyclopentan-2-yl)-2,3-dihydrobenzofuran-2-yl)methyl)morpholine (400 mg, yield 56%).

Step 4: Synthesis of N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)acetamide

The compound 4-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzo Furan-2-yl)methyl)morpholine (400 mg, 0.49 mmol) and N-benzyl-2-(5-bromopyridin-2-yl)acetamide (100 mg, 0.33 mmol) were added to ethylene glycol dimethyl ether (5 mL), then add sodium carbonate solution (2 mol/L, 0.49 mL), add tetrakis (triphenylphosphine) palladium (20 mg, 0.016 mmol) under nitrogen protection, and heat the reaction solution to 90°C and stir for 10 hours. The reaction solution was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (20 mL Liquid chromatography was used for separation. The separation method was (chromatographic column: Titank C18 10μ30mm×40cm; mobile phase: A=water+0.1% ammonia (99%), B=acetonitrile; gradient 35%-65%B, 13 minutes) , the compound N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)acetamide (223.3 mg, product rate 49%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.71–8.68(m,1H),8.61(t,1H),7.91(m,1H),7.55(d,1H),7.41(m,1H), 7.37(d,1H),7.34–7.19(m,5H),6.85(d,1H),5.07–4.97(m,1H),4.29(d,2H),3.69(s,2H),3.58(t, 4H),3.34(d,1H),3.30(t,1H),3.00(dd,1H),2.65(m,1H),2.58–2.50(m,2H),2.45(m,2H).

LC-MS,M/Z(ESI):444.4[M+H] ⁺

Example 2: N-benzyl-2-(5-(2-methyl-2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)ethane Preparation of amide (I-2)

The synthetic route of compound I-2 is as follows:

Step 1: Synthesis of 1-bromo-4-((2-methylallyl)oxo)benzene

Dissolve 4-bromophenol (36.8g, 213mmol) in N,N-dimethylformamide (600mL), add potassium carbonate (88.1g, 638mmol) and 3-chloro-2-methylprop-1-ene (23.1g, 255mmol), vacuumed, replaced with nitrogen three times, and stirred at 70°C for 8 hours. After the reaction is completed, water (600mL) is then added, and then extracted with petroleum ether (300*2mL). The organic phases are combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1-bromo-4-((2-methylallyl). Base)oxo)benzene (40g, crude product) was directly used in the next reaction.

¹ H NMR (400MHz, CDCl ₃ ): δ7.37(d,2H),6.81(d,2H),5.08(s,1H),5.00(s,1H),4.41(s,2H),1.83(s ,3H).

Step 2: Synthesis of 4-bromo-2-(2-methylallyl)phenol

Dissolve 1-bromo-4-((2-methylallyl)oxo)benzene (10.0g, 44.0mmol) in N-methylpyrrolidone (60mL) and vacuum Empty, replace with nitrogen three times, and stir at 190°C for 4 hours. After the reaction is completed, add water (600mL), then extract with ethyl acetate (100mL*2), combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate to obtain a crude product, which is then subjected to column chromatography on silica gel (petroleum ether:ethyl acetate). Ester = 50:1-10:1) was purified to obtain 1-bromo-4-((2-methylallyl)oxo)benzene (3.20 g, yield 32%).

¹ H NMR (400MHz, CDCl ₃ ): δ7.22-7.24(m,2H),6.72(d,1H),4.94(s,1H),4.85(s,1H),3.34(s,2H),1.74 (s,3H)

LC-MS,M/Z(ESI):225.0[M+H] ⁺

Step 3: Synthesis of (5-bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methanol

Dissolve 1-bromo-4-((2-methylallyl)oxo)benzene (2g, 8.81mmol) in dichloromethane (80mL), and slowly add 3-chloroperoxybenzoic acid ( 85%, 1.88g, 9.25mmol), the reaction solution was stirred at 0°C for 2 hours. Add saturated sodium bicarbonate solution (30 mL), separate the layers, place the organic phase in a round-bottomed flask, add p-toluenesulfonic acid monohydrate (167 mg, 0.88 mmol), and stir at room temperature for 0.5 hours. The reaction solution was washed with saturated sodium bicarbonate (20 mL), water (10 mL), and saturated brine (20 mL), then dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 5:1-1:1) to obtain (5-bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methanol. (1.39g, yield 65%).

Step 4: Synthesis of (5-bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methanesulfonate methyl ester

(5-Bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methanol (300 mg, 1.23 mmol) was dissolved in dichloromethane (5 mL), and methylsulfonyl chloride was added in sequence under ice bath (212mg, 1.85mmol) and triethylamine (162mg, 1.60mmol), the reaction solution was raised to room temperature and stirred for 4 hours. Add water (10 mL) to the reaction solution under ice bath, separate the layers, extract the aqueous phase with dichloromethane (5 mL*3), and combine the organic phases. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, then filtered and concentrated to obtain crude product (5-bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methanesulfonate. The ester (380 mg, yield 96%) was directly used in the next reaction.

Step 5: Synthesis of 4-((5-bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methyl)morpholine

(5-Bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methanesulfonate methyl ester (350mg, 1.09mmol) and morpholine (2g, 22.96mmol) were stirred at 120°C for 12 hours . The reaction solution was concentrated and separated and purified using a silica gel column (dichloromethane: methanol (V/V) = 20:1) to obtain 4-((5-bromo-2-methyl- 2,3-Dihydrobenzofuran-2-yl)methyl)morpholine (250 mg, yield 73%).

Step 6: 4-((2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3 -Synthesis of dihydrobenzofuran-2-yl)methyl)morpholine

4-((5-bromo-2-methyl-2,3-dihydrobenzofuran-2-yl)methyl)morpholine (100 mg, 0.320 μmol), pinacol diborate (122 mg, 480 μmol) , potassium acetate (94 mg, 961 μmol) and 1,1-bis(diphenylphosphine)ferrocene palladium dichloride (22.45 mg, 32 μmol) were added to 1,4-dioxane (3 mL), and nitrogen was replaced three times , and stirred at 90°C for 12 hours under nitrogen protection. The reaction solution was concentrated, water (10 mL) was added, extracted with ethyl acetate (5 mL*3), and the organic phases were combined. The organic phase was washed with saturated sodium chloride (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude product 4-((2-methyl-5-(4,4,5,5-tetramethyl-1, 3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-2-yl)methyl)morpholine (80 mg, yield 70%).

Step 7: N-benzyl-2-(5-(2-methyl-2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)ethane Synthesis of amides

4-((2-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzene Furan-2-yl)methyl)morpholine (80 mg, 223 μmol), N-benzyl-2-(5-bromopyridin-2-yl)acetamide (56.6 mg, 186 μmol), potassium carbonate (51.3 mg, 371 μmol) and 1,1-bis(diphenylphosphine)ferrocene palladium dichloride (13.58 mg, 19 μmol) were placed in 1,4-dioxane (1 mL) and water (0.7 mL), and replaced with nitrogen Three times, the reaction liquid was stirred at 100°C for 12 hours under the protection of nitrogen gas. The reaction solution was concentrated and purified by thin layer chromatography on silica gel plate (dichloromethane:methanol (V/V)=10:1) to obtain N-benzyl-2-(5-(2-methyl-2-( Morpholinemethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)acetamide (35 mg, yield 34%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.71(d,1H),8.65–8.59(m,1H),7.92(dd,1H),7.54(s,1H),7.42–7.35(m,2H ),7.34–7.21(m,5H),6.79(d,1H),4.30(d,2H),3.70(s,2H),3.58–3.44(m,4H),3.28(s,2H),2.57( d,2H),2.55–2.51(m,2H),2.48–2.42(m,2H),1.38(s,3H).

LC-MS,M/Z(ESI):458.3[M+H] ⁺

Example 3: (R)-N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)acetamide Preparation

Preparation of (S)-N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridin-2-yl)acetamide

The racemic compound I-1 (200 mg) was separated by SFC. The separation method ("%" in the separation method is volume percentage) is:

Condition 1: Chromatographic column: Chiralcel OD-3 50×4.6mm ID, 3μm; mobile phase A is supercritical fluid CO ₂ (Phase A for CO ₂ ), mobile phase B is MeOH+ACN (0.05% DEA) (Phase B) for MeOH+ACN(0.05%DEA), its volume ratio is 2:1); gradient: supercritical fluid carbon dioxide containing 40% mobile phase B (40%MeOH+ACN(0.05%DEA) in CO ₂ ); flow rate: 3mL/min; detector: PDAColumn; column temperature: 35°C; column pressure: 100Bar.

Condition 2: Chromatographic column: Chiralcel OD-3 50×4.6mm I.D., 3μm; mobile phase A: supercritical fluid carbon dioxide, mobile phase B: ethanol (0.05% diethylamine); gradient elution: containing ethanol (0.05% diethylamine) Ethylamine) supercritical fluid carbon dioxide, its proportion is from 5% to 40%; flow rate: 3mL/min; detector: PDAColumn; column temperature: 35°C; column pressure: 100Bar.

Peak 1 obtained by resolution was concentrated to obtain compound (I-1A)(R)-N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5- yl)pyridin-2-yl)acetamide or (S)-N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridine -2-yl)acetamide (55.6 mg, 99.81%ee, retention time: t=1.204 min).

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.75–8.71(m,1H),8.70(t,1H),7.91(m,1H),7.55(d,1H),7.41(m,1H), 7.37(d,1H),7.34–7.19(m,5H),6.85(d,1H),5.07–4.97(m,1H),4.29(d,2H),3.69(s,2H),3.58(t, 4H),3.34(d,1H),3.30(t,1H),3.00(dd,1H),2.65(m,1H),2.58–2.50(m,2H),2.45(m,2H)

LC-MS,M/Z(ESI):444.2[M+H] ⁺

Peak 2 obtained by separation was concentrated to obtain compound (I-1B)(S)-N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5- yl)pyridin-2-yl)acetamide or (R)-N-benzyl-2-(5-(2-(morpholinmethyl)-2,3-dihydrobenzofuran-5-yl)pyridine -2-yl)acetamide (58.6 mg, 99.68%ee, retention time: t=2.010 min).

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.74–8.71(m,1H),8.70(t,1H),7.91(m,1H),7.55(d,1H),7.41(m,1H), 7.37(d,1H),7.34–7.19(m,5H),6.85(d,1H),5.07–4.97(m,1H),4.29(d,2H),3.69(s,2H),3.58(t, 4H),3.34(d,1H),3.30(t,1H),3.00(dd,1H),2.65(m,1H),2.58–2.50(m,2H),2.45(m,2H).

LC-MS, M/Z(ESI): 444.2[M+H] ⁺ .

For the preparation method of the following compounds, refer to Example 1:

The control compound (trade name: Tirbanibulin) in the test example of the present invention is a dual-action Src kinase and tubulin polymerization inhibitor, and has been approved by the FDA and the European Union for the topical treatment of actinic keratosis on the face or scalp. . The preparation of the control compound refers to patent WO 2008/002676 A2. The structure of the control compound is as follows:

Test Example 1: Inhibition of tubulin monomer polymerization test

Use Tubulin Polymerization Assay Kit (cytoskeleton, Cat.#BK011P) to carry out compound inhibition of tubulin monomer. Bulk polymerization test.

Before the test, place the 96-well plate (Corning Costar, Cat.#3686) supplied with the kit in a microplate reader (MD, SpectraMax M5) and heat it to 37°C for 10 minutes. Take out the 96-well plate and add 5 μl of 10× compound solution or blank solution, and then put the 96-well plate into the microplate reader and incubate it at 37°C for 1 min to heat the compound solution to 37°C. Take out the 96-well plate and quickly add 50 μl of the reaction mixture configured according to the supplier's instructions to each well for 1 min. Complete the sample addition within 3 seconds to avoid bubbles, then immediately put the 96-well plate into the microplate reader, shake for 5 seconds, and use Kinetic mode to detect continuously for 30min-60min at 37°C under the conditions of excitation light 360nm and emission light 420nm, and detect once every 30s. , taking the detection time as the X-axis and the fluorescence signal value as the Y-axis, the tubulin monomer polymerization curve is obtained. The larger the Vmax value and the higher the maximum fluorescence signal value of the polymerization curve, the lower the inhibitory efficiency of the compound.

The results of the compound's inhibition of tubulin polymerization test show that the compound of the present invention can significantly inhibit the polymerization of tubulin monomers and has high inhibitory activity.

Test Example 2: Cell Proliferation Inhibition Test

Human skin squamous cell carcinoma cell A-431 (ATCC, CRL-1555), human embryonic kidney epithelial cell 293T (China Type Culture Collection Center, GDC0187) and mouse embryonic fibroblast 3T3-Swiss (ATCC, CCL- 92) Proliferation test detects the inhibitory effect of small molecule compounds on cell proliferation.

A-431 cells, 293T cells and 3T3-Swiss cells were cultured in DMEM medium containing 10% fetal calf serum and grown in a 37°C, 5% _CO2 incubator. Logarithmic phase cells were seeded into a 96-well cell culture plate at 1000 cells/well, 100 μL per well, and placed in a 37-cell, 5% CO ₂ incubator for overnight culture. On the next day, add 100 μL of gradient diluted 2× solution of the compound to be tested to each well, DMSO as a positive control, and 10 μM staurosporine (Aladdin, S102392) as a negative control group. The culture plate after adding the compounds continues to be incubated for 37 seconds. , incubate in a 5% _CO2 incubator for 4 days. Use after incubation is complete luciferase assay system (Promega, G9243) and measure the fluorescence signal value on Envision 2104 Multilabel Reader according to the instructions provided by the supplier. Calculate the inhibition rate by the following formula, and then draw a curve using the log value of the inhibitor concentration as the X-axis and the inhibition rate as the Y-axis. Use Graphpad 8.0 to calculate the IC ₅₀ .

Inhibition%=(positive control group signal-test hole signal)/(positive control group signal-negative control group signal)*100

Table 1 Inhibitory results of test compounds on A431 cell proliferation activity

The results of the compound's inhibition of cell proliferation test show that the compound of the present invention can significantly inhibit cell proliferation.

Test Example 3: Pharmacokinetic Test

For the mouse pharmacokinetics test, 3 male ICR mice, 20-25g, were fasted overnight and administered via tail vein injection (1mg/kg or 5mg/kg). Blood was collected before administration and at 15, 30 minutes and 1, 2, 4, 8, and 24 hours after administration. Blood samples were centrifuged at 6800 g and 2-8°C for 6 minutes, and plasma was collected and stored at -80°C. Take the plasma at each time point, add 3-5 times the amount of acetonitrile solution containing the internal standard, mix, vortex and mix for 1 minute, centrifuge at 13000 rpm for 10 minutes at 4°C, take the supernatant, add 3 times the amount of water, mix, and take an appropriate amount of the mixture. Perform LC-MS/MS analysis. The main pharmacokinetic parameters were analyzed using WinNonlin 7.0 software non-compartmental model.

Table 2 Mouse pharmacokinetic test results

The mouse pharmacokinetic test results show that the compound of the present invention exhibits pharmacokinetic properties consistent with topical administration to the skin, has fast metabolism in the body, low potential toxicity, and good drug potential.

Test Example 4: Skin Irritation Test

Use a rabbit skin irritation test to determine the skin irritation of a compound. Specifically, monotriglyceride was used as a blank preparation, and the control compound and compound 1-1A were prepared into test samples using monotriglyceride. At D0, the hair on the left and right sides of the rabbit's abdomen was removed. Use a cotton swab to apply the blank preparation and the test substance sample to the left skin preparation area at a dose of 0.05mL/cm ² . Apply the same dose of the blank preparation to the right side. Apply evenly and cover and fix with paper tape. The application time is 6 hours. , wash off the agent with clean water after 6 hours. Each trial was run in 6 parallels. Administer for 5 consecutive days. Observe the skin reaction with the naked eye under natural light or full-spectrum light before administration and 24h (D1), 48h (D2), 72h (D3), 96h (D4), and 120h (D5) after administration, and score according to the following standards for scoring.

Skin irritation reaction scoring criteria

The results of the skin irritation test are shown in Figure 1. The total score of the irritation reaction in the figure is the sum of the erythema score and the edema score. The results show that the compound of the present invention exhibits low skin irritation reaction, low potential toxicity, and good medicinal properties.

Test Example 5: Compound inhibits Src signaling pathway

The Western Blot method was used to detect the compound's inhibition of p-SRC on A-431 skin cancer cells (ATCC, CRL-1555) to evaluate the inhibitory effect of the compound on the SRC signaling pathway.

A-431 cells were cultured in DMEM medium containing 10% fetal calf serum and placed in a 37°C, 5% CO2 incubator. 500,000 cells/well were seeded in a 12-well cell culture plate and cultured overnight in a 37°C, 5% CO2 incubator. The next day, the culture medium was replaced, and different concentrations of compounds were added. DMSO was used as a control. The culture plate after adding the compounds was placed in a 37°C, 5% CO ₂ incubator to continue culturing for 24 hours. Wash the cells once with PBS, lyse the cells with RIPA lysis buffer (R0010, Soleba) at low temperature for 30 minutes, then collect the protein lysate into a centrifuge tube, centrifuge at 14000g for 10 minutes at 4°C, and transfer the supernatant to fresh Tube. BCA protein concentration assay kit (P0009, Beyotime Biotechnology) was used to detect and calculate the protein concentration. Use 5× protein loading buffer to quantify the protein of each sample consistently, boil at 100°C for 5 minutes, and perform western blot to detect p-SRC levels. p-SRC antibody (Ab185617) was purchased from Abcam Company, and GAPDH antibody (60004-1-Ig ) purchased from proteintech company.

Test results show that the compound of the present invention can significantly inhibit the phosphorylation of SRC, has excellent p-SRC inhibitory activity, and can block the SRC downstream signaling pathway.

Although the embodiments of the present invention have been shown and described above, it can be understood that the above-mentioned embodiments are illustrative and should not be construed as limitations of the present invention. Those of ordinary skill in the art can make modifications to the above-mentioned embodiments within the scope of the present invention. The embodiments are subject to changes, modifications, substitutions and variations.

Claims (15)

1. A compound represented by formula I, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug:
   

   in,

   Ring B is a saturated, unsaturated or partially saturated 5-6 membered heterocyclic ring;

   Y is a C ₁ -C ₆ alkyl group that is unsubstituted or substituted by one or more R ¹ ; the R ¹ is selected from halogen, hydroxyl, amino, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl and C ₁ -C ₆ alkoxy group; when R ¹ is multiple, the R ^1s are the same or different;

   Z is C ₃ -C ₈ cycloalkyl, 3-11 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl, wherein the C ₃ -C ₈ cycloalkyl, 3- 11-membered heterocycloalkyl, 6-10-membered aryl, 5-10-membered heteroaryl are optionally substituted by one or more R ² ; when R ² is multiple, the R ² is the same or different;

   R ² is each independently selected from halogen, hydroxyl, amino, cyano, carbonyl, oxo (=O), carboxyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl , C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkyl-OH, C ₁ -C ₆ alkyl -C(=O)-, C ₁ -C ₆ alkylcyano, C ₁ -C ₆ alkyl Oxygen, C ₁ -C ₆ haloalkoxy, C ₁ -C ₆ alkoxycarbonyl, C ₁ -C ₆ alkoxy C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy -C(=O )-C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ halocycloalkyl, C ₃ -C ₈ cycloalkylcarbonyl, C ₃ -C ₈ cycloalkyl -C ₁ -C ₆ alkyl-carbonyl, C ₃ -C ₈ cycloalkoxy, C ₃ -C ₈ halocycloalkoxy, optionally aminocarbonyl with 1 or 2 C ₁ -C ₆ alkyl groups Groups, C ₁ -C ₆ alkylsulfonyl groups, aminosulfonyl groups optionally having 1 or 2 C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkylsulfonylamino groups and any Choose an amino group with 1 or 2 C ₁ -C ₆ alkyl groups;

   Q is a 6-10-membered aryl group that is unsubstituted or substituted by one or more R ³ , or a 5-10-membered heteroaryl group that is unsubstituted or substituted with one or more R ³ ;

   The heteroatoms of the heteroaryl group are N, O or S, and the number of heteroatoms is 1, 2 or 3;

   Each of the R ³ is independently selected from halogen, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ halocycloalkyl, C ₃ -C ₈ cycloalkoxy and C ₃ -C ₈ halocycloalkoxy; or when R ³ is multiple, substituted 6-10 membered aryl group Or a 5-10 membered heteroaryl group and R ³ together form an O-containing saturated heterocyclic ring;

   R ^a , R ^b and R ^c are absent, or are each independently halogen, hydroxyl, cyano, carbonyl, oxo (=O), C ₂ -C ₆ alkenyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkyl -OH, C ₁ -C ₆ alkyl -C(=O)-, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ halocycloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy, -COO-C ₁ -C ₆ alkyl or -C(O)NR ⁴ R ⁵ ; wherein, The R ⁴ and R ⁵ are each independently hydrogen or C ₁ -C ₆ alkyl;

   m, n, p, q are independently 0, 1, 2 or 3.
2. The compound represented by formula I according to claim 1, its tautomer, its stereoisomer, its hydrate, and its solvation A substance, a pharmaceutically acceptable salt thereof or a prodrug thereof, characterized in that it meets one or more of the following conditions:

   (1) The heteroatoms in each heterocycle, each heteroaryl group and each heterocycloalkyl group are independently selected from one of N, O and S, and the number can be 1, 2 or 3;

   Preferably, when the heteroatom contains an S atom, the lone pair of electrons on the S atom is not replaced by an O atom to form

   (2) The O-containing saturated heterocyclic ring may be a 3-11 membered O-containing saturated heterocyclic ring;

   (3) In Ring B, the saturated, unsaturated or partially saturated 5-6-membered heterocyclic ring is a 5-6-membered heteroaromatic ring or a 5-6-membered heteroolefin ring, and the heteroatoms are selected from N, O and one or more of S, the number is 1, 2 or 3;

   (4) In Y, the C ₁ -C ₆ alkyl group in the unsubstituted or substituted C ₁ -C ₆ alkyl group with one or more R ¹ is methyl, ethyl, n-propyl, isopropyl , n-butyl, isobutyl, sec-butyl or tert-butyl;

   (5) In Z, the 3-11 membered heterocycloalkyl group is a 5, 6, 7 or 8 membered saturated monocyclic heterocyclic ring, a 6, 7, 8, 9 or 10 membered saturated spiro ring or bridge Ring heterocycle, or 8, 9 or 10-membered saturated cyclic heterocycle, with 1 to 3 heteroatoms selected from N, O and S, preferably 5, 6, 7 or 8-membered saturated monocyclic ring The heterocycle is further preferably morpholinyl, 1,4-oxepanyl, azetidinyl, pyrrolidinyl or piperazinyl, such as morpholinyl;

   (6) In R ² , the halogen is fluorine, chlorine or bromine, such as fluorine;

   (7) In R ² , the C ₁ -C ₆ alkyl group in the C 1 -C 6 alkyl group and _the C ₃ -C ₈ cycloalkyl- _{C 1 -C 6} alkyl-carbonyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl;

   (8) In R ² , the C ₁ -C ₆ alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, such as methoxy;

   (9) In R ² , the C ₃ -C ₈ cycloalkyl group in the C ₃ -C ₈ cycloalkyl-C ₁ -C ₆ alkyl-carbonyl group is cyclopropyl, cyclobutyl, or cyclopentane. base or cyclohexyl group, such as cyclopropyl group;

   (10) In Q, the 6-10-membered aryl group in the 6-10-membered aryl group that is unsubstituted or substituted by one or more R ³ is phenyl or naphthyl, such as phenyl;

   (11) In Q, the 5-10-membered heteroaryl group that is unsubstituted or substituted by one or more R ³ is a 5-6-membered heteroaryl group, and the 5-6-membered heteroaryl heteroatom is N , one or more of O or S, the number of heteroatoms is 1, 2 or 3;

   (12) In R ³ , the halogen is fluorine, chlorine or bromine, such as fluorine;

   (13) In R ^a , the halogen is fluorine, chlorine or bromine, such as fluorine or chlorine;

   (14) In R ^a , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl ;

   (15) In R ^a , the C ₂ -C ₆ alkynyl group is ethynyl, propynyl or butynyl, for example

   (16) In R ^c , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl .
3. The compound represented by formula I according to claim 1, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug, characterized in that , which satisfies one or more of the following conditions:

   (1) The ring B is a saturated, unsaturated or partially saturated 5-6 membered heterocyclic ring containing 1, 2 or 3 heteroatoms;

   Preferably, the heteroatoms are selected from heteroatoms of N, O, and S; when there are multiple heteroatoms, the heteroatoms are the same or different;

   More preferably, the saturated, unsaturated or partially saturated 5-6 membered heterocyclic ring is a 5-6 membered heteroaromatic ring or a 5-6 membered heteroolefin ring, and the heteroatoms are selected from N, O and S. One or more kinds, the number is 1, 2 or 3;

   (2) The Y is selected from unsubstituted or methyl-substituted C ₁ -C ₆ alkyl;

   (3) Z is a 3-11 membered saturated heterocyclic ring that is unsubstituted or substituted by R ² ; said R ² is C ₁ -C ₆ alkyl or C ₃ -C ₈ cycloalkyl-C ₁ -C ₆ alkyl -Carbonyl group, the substituents are 1 or 2; when the substituents are 2, the substituents are the same or different;

   Preferably, the 3-11-membered saturated heterocyclic ring has 1 to 3 heteroatoms selected from N, O and S; and/or the 3-11-membered saturated heterocyclic ring is a monocyclic ring, a paracyclic ring, or a spirocyclic ring. or bridge ring;

   More preferably, Z is a 5-8-membered monocyclic ring, a 6-10-membered spirocyclic ring, a 6-10-membered bridged ring or an 8-10-membered paracyclic ring that is unsubstituted or substituted by R ² ;

   (4) The Q is selected from an unsubstituted or substituted phenyl group with R ³ , a 5-6 membered heteroaryl group that is unsubstituted or substituted with R ³ ; the heteroatom of the heteroaryl group is N, O or S, The number of heteroatoms is 1-3; the R ³ substitution is one or more substitutions, and each R ³ is independently selected from hydrogen, halogen, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy; or, when R ³ is multiple, the substituted phenyl or 5-6-membered heteroaryl and R ³ together form an O-containing saturated heterocyclic ring;

   Preferably, Q is selected from unsubstituted or phenyl substituted by R ³ ;

   More preferably, Q is selected from unsubstituted or fluorine-substituted phenyl;

   (5) R ^a does not exist, or is independently halogen, hydroxyl, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl, C ₂ -C ₆ alkynyl or C ₁ -C ₆ haloalkyl;

   Preferably, R ^a does not exist, or is independently halogen, cyano, C ₁ -C ₆ alkyl or C ₂ -C ₆ alkynyl;

   (6) R ^b does not exist, or is independently halogen, hydroxyl, cyano, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl, C ₂ -C ₆ alkynyl or C ₁ -C ₆ haloalkyl;

   Preferably, R ^b does not exist, or is independently halogen, cyano or C ₁ -C ₆ alkyl;

   (7) R ^c does not exist, or is independently halogen, hydroxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ deuterated alkyl, C ₁ -C ₆ haloalkyl or C ₁ -C ₆ alkoxy base;

   Preferably, R ^c is absent, or is independently halogen or C ₁ -C ₆ alkyl.
4. The compound represented by formula I according to claim 3, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug, characterized in that , which satisfies one or more of the following conditions:

   (1) Ring B is dihydrofuran, furan, dihydrothiophene, thiophene, pyrrole, dihydropyrrole, imidazole, dihydroimidazole, oxazole, dihydroxazole, thiazole, dihydrothiazole, piperidine, tetrahydropyridine , morpholine, piperazine, hexahydropyrimidine, ectoine, triazine, tetrahydrotriazine, tetrahydropyran, dihydropyran, tetrahydrothiopyran or dihydrothiopyran;

   Preferably ring B is dihydrofuran, furan, dihydrothiophene or thiophene; for example

   (2)Y is methyl;

   (3) Z is

   Preferably, Z is

   (4)Q is

   (5) R ^a does not exist, or is independently fluorine, chlorine, cyano, methyl or propynyl;

   (6) R ^b does not exist or is fluorine;

   (7) R ^c does not exist or is methyl.
5. The compound represented by formula I according to any one of claims 1 to 4, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its Prodrug, characterized by, for
6. The compound represented by formula I according to claim 1, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug, characterized in that , the compound represented by formula I has a structure represented by formula I-1:
   

   where the dotted line represents an optional double bond;

   When the dotted line does not represent a double bond, X ₁ and X ₂ are independently C, NH, O or S; at least one of X ₁ and X ₂ is not C;

   When the dotted line represents a double bond, X ₁ is C and X ₂ is NH, O or S;

   Ra, ^Rb , ^Rc , Y, Z, Q, m, n, p and q are as ^defined in claim 1.
7. The compound represented by formula I according to claim 1, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug, characterized in that , the compound represented by formula I has a structure represented by formula I-1a, formula I-1b, formula II-1, formula II-2 or formula II-3:
   

   Wherein, Ra, ^{Rb , Rc} , Y, Z, Q, m, n, p and q are defined as in claim 1.
8. The compound represented by formula I as claimed in claim 7, its tautomer, its stereoisomer, its hydrate, and its solvation A substance, a pharmaceutically acceptable salt thereof or a prodrug thereof, characterized in that the structure represented by formula II-1 has a structure represented by formula II-1a, II-1b or II-1c:
   

   In formula II-1c, n is 0 or 1; p is 0 or 1; Ra and Rc are each independently methyl, fluorine, chlorine or cyano;

   Preferably, in formulas II-1a and II-1b, R ^c is absent or C ₁ -C ₆ alkyl; Z is morpholinyl; Q is phenyl;

   Preferably, for

   Preferably, R ^b does not exist;

   Preferably, the structure represented by formula II-1c has the structure represented by formula II-1c-1 or II-1c-2
   
9. The compound represented by formula I according to claim 1, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug, characterized in that , the compound represented by formula I is selected from any of the following compounds:
   
   
10. The compound represented by formula I according to claim 9, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug, characterized in that ,

    The compound represented by formula I is
    

    Or, the compound represented by formula I is:

    The compound that peaks first in chiral separation or chiral analysis;

    or, Compounds that elute late in chiral separation or chiral analysis;

    The detection parameters of the chiral separation or chiral analysis are preferably the following detection parameters 1 or 2:

    Detection parameter 1: chromatographic column: Chiralcel OD; mobile phase A: CO ₂ , mobile phase B: MeOH and acetonitrile containing 0.05% diethylamine;

    More preferably, the separation gradient: supercritical fluid carbon dioxide of a mixed solution of 40% MeOH and acetonitrile containing 0.05% diethylamine, the volume ratio of MeOH to acetonitrile is 2:1; flow rate: 3mL/min; detector: PDAColumn; Column temperature: 35℃; column pressure: 100Bar;

    Detection parameter 2: Chromatographic column: Chiralcel OD-3 50×4.6mm I.D., 3μm; mobile phase A: supercritical fluid carbon dioxide, mobile phase B: ethanol containing 0.05% diethylamine; retention of the compound that peaks first The time is 1.204min; the retention time of the compound that peaks later is 2.010min;

    More preferably, gradient elution: supercritical fluid carbon dioxide containing 0.05% diethylamine in ethanol, with a ratio from 5% to 40%; flow rate: 3mL/min; detector: PDAColumn; column temperature: 35°C; column pressure :100Bar.
11. A pharmaceutical composition, characterized in that the pharmaceutical composition includes: the compound represented by formula I according to any one of claims 1-10, its tautomer, and its stereoisomer, Its hydrate, its solvate, its pharmaceutically acceptable salt or its prodrug; and a pharmaceutically acceptable carrier.
12. The use of a substance M, characterized in that the substance M is a compound represented by formula I according to any one of claims 1 to 10, its tautomers, and its stereoisomers, Its hydrate, its solvate, its pharmaceutically acceptable salt, its prodrug or the pharmaceutical composition of claim 11; the use is:

    Inhibit tubulin polymerization or Src kinase;

    and/or, prevent and/or treat tubulin polymerization-mediated or Src kinase-related diseases;

    and/or, preparing drugs, pharmaceutical compositions or preparations for inhibiting tubulin polymerization or Src kinase, and/or preventing and/or treating tubulin polymerization-mediated or Src kinase-related diseases;

    Or, prepare a medicament for treating and/or preventing actinic keratosis.
13. The use according to claim 12, wherein the tubulin polymerization-mediated or Src kinase-related diseases include: one, two or more of tumors, skin diseases and/or other diseases;

    The tumors preferably include: solid tumors, sarcomas, hematological cancers, subtypes include breast cancer, ovarian cancer, prostate cancer, cervical cancer, testicular cancer, colon cancer, colorectal cancer, liver cancer, non-small cell lung cancer, squamous cell carcinoma cell carcinoma, small cell lung cancer, gastric cancer, gastrointestinal stromal tumor, pancreatic cancer, bladder cancer, germ cell tumor, mastocytoma, mastocytosis, glioblastoma, neuroblastoma, astrocytoma , melanoma, B-cell lymphoma, T-cell lymphoma, slowly progressive lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, Chronic lymphocytic leukemia, myeloma, and/or myelodysplastic syndrome;

    The skin diseases preferably include: actinic keratosis, psoriasis, atopic dermatitis, psoriasis, vitiligo, roseola and/or systemic lupus erythematosus; the actinic keratosis is preferably scaly. skin cancer;

    The other diseases preferably include: autoimmune diabetes, diabetic retinopathy, liver fibrosis, pulmonary fibrosis, renal fibrosis, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinocerebellar Degenerative diseases, atherosclerosis, anemia, sickle cell anemia, thalassemia, osteoarthritis, rheumatoid arthritis, malaria, trypanosomiasis, helminthiasis, protozoal infections, multiple sclerosis, lupus , asthma, allergic rhinitis and/or inflammatory bowel disease;

    The actinic keratosis is preferably squamous skin cancer.
14. A compound represented by Formula I according to any one of claims 1 to 10, its tautomer, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt or its The preparation method of prodrug is characterized in that it includes the following steps:

    In the presence of an alkaline reagent and a palladium catalyst, compound II and compound II are subjected to a Suzuki reaction in a solvent to obtain the compound represented by formula I;
    

    _Wherein ^{, _ _}
15. A compound represented by formula II or a compound represented by formula IV:
    

    _Wherein ^{, _}

    The compound represented by formula II is preferably

    The compound represented by formula IV is preferably