WO2017181974A1 - Five-membered heterocyclic compound, preparation method therefor, pharmaceutical composition and use - Google Patents

Abstract

Provided are a five-membered heterocyclic compound, a preparation method therefor, a pharmaceutical composition and a use. Specifically, provided is a compound having formula (I), the definitions of the various groups being laid out in the description. The compound having formula (I) has an effect of inhibiting tubulin activity, and may be used to prepare a drug treating or preventing mammalian diseases related to tubulin regulation abnormalities.

Description

Five-membered heterocyclic compound, preparation method thereof, pharmaceutical composition and use thereof Technical field

The invention relates to the field of biomedicine, in particular to a multi-substituted five-membered heterocyclic compound, a preparation method thereof, a pharmaceutical composition and use thereof.

Background technique

Microtubules are composed of tubulin, which are found in the cytoplasm of all eukaryotes. They play an important cytoskeleton role in cells, maintain cell morphology, assist intracellular transport, and assemble into spindles, granules, and other proteins. Central granules, flagella, ciliated neural tube and other structures. Microtubule-associated proteins bind to microtubules and regulate microtubule function, including MAP1, MAP12, MAP4, tau, and the like. Microtubules play a key role in cell mitosis and chromosome segregation and can affect tumor cell proliferation as a target for antitumor drugs. Tubulin inhibitors can effectively prevent mitosis of tumor cells and cause cells to enter apoptotic phase, thereby inhibiting tumor growth.

In the nervous system, the stability of the microtubule system is also the basis for maintaining nutrient transport between the cell bodies and the processes. Tubulin and microtubule-associated proteins have been shown to be closely associated with key proteins in neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Therefore, there is an urgent need in the art to develop novel tubulin modulators.

Summary of the invention

It is an object of the present invention to provide a novel tubulin modulator.

In a first aspect of the invention, there is provided a use of a compound of formula (I), an isomer, a racemate, a pharmaceutically acceptable salt, a crystalline hydrate, a solvate or a mixture thereof, characterized in that In order to (a) prepare a drug for treating or preventing a mammalian disease associated with abnormality of tubulin; (b) inhibit microtubule polymerization activity; (c) inhibit tumor cell growth:

among them:

X _{1 is} selected from N or CR ₁ (when R1 is H, denoted as C);

X _{2 is} selected from NR ₁ , O or S;

X _{3 is} selected from the group consisting of NH, O, S, C=O, C=S, C=NH, -(C=O)-NH-, -(C=O)-O-,-(C=O )-S-,-(C=S)-NH-,-(C=S)-O-,-(C=S)-S-,-(C=NH)-NH-,-(C=NH )-O- or -(C=NH)-S-;

R _{1 is} selected from the group consisting of hydrogen, hydrazine, halogen, amino, hydroxy, nitro, cyano, carboxy, C _2-6 ester, C _1-6 amide, unsubstituted or halogenated C _1-12 alkane Or cycloalkyl, -CH ₂ -Y-(C _1-12 alkyl or cycloalkyl) (wherein Y is O or NH or S), C _1-12 aryl or heteroaryl, -CH ₂ - (C _1-12 aryl or heteroaryl);

The Ar ₁ group is a substituted or unsubstituted C _1-18 aryl or heteroaryl group, a substituted or unsubstituted -CH ₂ -(C _1-12 aryl or heteroaryl), substituted or unsubstituted C _{1 -18} heterocyclic group, substituted or unsubstituted -CH ₂ -(C _1-12 heterocyclic group);

The Ar ₂ group is hydrogen, substituted or unsubstituted C _1-12 alkyl or cycloalkyl, substituted or unsubstituted -CH ₂ -Y-(C _1-12 alkyl or cycloalkyl) (where Y is O or NH or S), substituted or unsubstituted C _1-12 aryl or heteroaryl, substituted or unsubstituted -CH ₂ -(C _1-12 aryl or heteroaryl), substituted or unsubstituted C _1-12 heterocyclic group;

m is selected from 0, 1, 2, 3, 4, 5, 6;

n is selected from 0, 1, 2, 3, 4, 5, 6;

Wherein the substitution is substituted with one or more substituents selected from the group consisting of halogen, amino, hydroxy, nitro, cyano, trifluoromethyl, C _1-12 alkyl or cycloalkyl, a C _1-12 alkoxy group, an oxygen atom (ie, =0), a C _1-12 alkylamino group unsubstituted or substituted with a C _1-4 alkylamino group, a C _2-6 ester group, a C _2-6 acyl group, C _1-6 amido, thio C _1-12 alkyl group, a substituted carboxyl group, unsubstituted or substituted with 1-5 halogen, amino, hydroxy, nitro, cyano, trifluoromethyl C _1-12 aryl group Or a heteroaryl group, or a C _1-12 heterocyclic group which is unsubstituted or substituted by 1 to 5 halogen, amino, hydroxy, nitro, cyano, trifluoromethyl (containing 1-5, preferably 1-3 a hetero atom selected from N, O or S).

In another preferred embodiment, the X _{3 is} selected from the group consisting of NH, O, S, C=O, C=S, C=NH, -(C=O)-NH-, -(C=O) -O-,-(C=O)-S-,-(C=S)-NH-,-(C=S)-O-,-(C=S)-S-;

The R _{1 is} selected from the group consisting of hydrogen, hydrazine, halogen, amino, hydroxy, nitro, cyano, carboxy, ester, amide, unsubstituted C _1-6 alkyl or cycloalkyl, (1 3) fluoro C _1-6 alkyl or cycloalkyl, (1-3) C _1-6 amine substituted C _1-6 alkyl or cycloalkyl, (1-3) C _1-6 alkoxy Substituted C _1-6 alkyl or cycloalkyl, -CH ₂ -Y-(C _1-6 alkyl), C _1-12 aryl or heteroaryl, -CH ₂ -(C _1-12 aryl or Heteroaryl).

In another preferred embodiment, in the Ar ₁ or Ar ₂ group, the C _5-12 aryl or heteroaryl group is selected from the group consisting of:

m is 0, 1, 2, 3, 4, 5;

n is 0, 1, 2, 3, 4, 5.

In another preferred embodiment, X ₁ , X ₂ , X ₃ , R ₁ , Ar ₁ , Ar ₂ , m and n are each independently a corresponding group corresponding to the compound 1-630.

In another preferred embodiment, the Ar _{1 is} selected from the group consisting of C _1-8 heteroaryl and C _2-10 heteroaryl, C _4-8 heteroaryl and C _2-10 heterocyclic.

In another preferred embodiment, the C _1-8 heteroaryl contains 1-3 nitrogen atoms.

In another preferred embodiment, the C _1-8 heteroaryl group is a six-membered ring.

In another preferred embodiment, the C _2-10 heteroaryl or C _2-10 heterocyclic group contains 1-3 nitrogen atoms.

In another preferred embodiment, the C _2-10 heteroaryl or C _2-10 heterocyclic group is a six-membered ring.

In another preferred embodiment, the heterocyclic group is a saturated or partially unsaturated heterocyclic group, and the heterocyclic group is a non-aromatic group.

In another preferred embodiment, the compound is Compound 1 to Compound 630.

In another preferred embodiment, the mammalian disease associated with dysregulation of microtubule-associated proteins is a disease selected from the group consisting of cancer, neurodegenerative diseases, malaria, AIDS, gout, diabetes.

In another preferred embodiment, the cancer is selected from the group consisting of colon cancer, cervical cancer, breast cancer, liver cancer, stomach cancer, kidney cancer, Lung cancer, fibrosarcoma, epidermal squamous cell carcinoma, prostate cancer, leukemia, pancreatic cancer, oral cancer, glioblastoma, neuroblastoma.

In another preferred embodiment, the tumor cell is a tumor cell selected from the group consisting of colon cancer cells, cervical cancer cells, breast cancer cells, liver cancer cells, gastric cancer cells, kidney cancer cells, lung cancer cells, and fibrosarcoma cells. Epidermal squamous cell carcinoma, prostate cancer cell, leukemia cell, pancreatic cancer cell, oral cancer cell, glioblastoma cell, neuroblastoma cell, paclitaxel-resistant lung cancer cell, vincristine-resistant oral cancer cell, resistance to melon Chronic myeloid leukemia cells.

In another preferred embodiment, the tumor cells are selected from the group consisting of HCT116, Hela, MCF-7, LM3, NCI-N87, Caki-1, A549, HT1080, A431, PC3, HL60, Panc-1, KB. , U87-MG, K562, Kasumi-1, THP-1, Jurkat, REH, Raji, RNK-16, KMS-1, P39, U118-MG, H4, SK-N-SH, SH-SY5Y, A549/Taxol , KB/VCR, K562/Adr.

In another preferred embodiment, the mammalian disease associated with dysregulation of microtubule-associated proteins is a disease selected from the group consisting of lymphoma, lung cancer, gastric cancer, pancreatic cancer, breast cancer, prostate cancer, leukemia, brain tumor, and Cervical cancer.

In a second aspect of the invention, there is provided a process for the preparation of a compound of formula (I):

Wherein X ₁ , X ₂ , X ₃ , R ₁ , Ar ₁ , Ar ₂ , m and n are as defined in the first aspect of the invention;

The method comprises the steps of:

(i-a) reacting a compound of formula a with a compound of formula b in an inert solvent to provide a compound of formula c;

(ii-a) reacting a compound of formula c with a compound of formula d in an inert solvent to provide a compound of formula e;

And when Q = NR ₁ , further comprising the optional step of: (iii-a) deprotecting a compound of formula e in an inert solvent to provide a compound of formula f;

Wherein X is a halogen; U is selected from the group consisting of halogen, OMs, OTs, boric acid, and pinacol borate; Q is O, S or NR ₁ ; P is selected from the group consisting of p-methoxybenzyl, benzyl. , tert-butoxycarbonyl;

Or the method comprises the steps of:

(i-b) in an inert solvent, the compound of formula g is substituted with a halogen to give a compound of formula h;

Wherein R ₁ is halogen (fluorine, chlorine, bromine or iodine)

The definitions of the remaining groups are the same as the first aspect of the invention.

In a third aspect of the invention, there is provided a process for the preparation of a compound of formula (I):

Wherein X ₁ , X ₂ , X ₃ , R ₁ , Ar ₁ , Ar ₂ , m and n are as defined in the first aspect of the invention;

The method comprises the steps of:

(i-c) a compound of formula a is subjected to a ring closure reaction with a compound of formula b in an inert solvent to provide a compound of formula c;

Preferably, the ring closure reaction is carried out by inorganic salt catalysis;

Wherein V is halogen, OMs, or OTs; the remaining groups are as defined in the first aspect of the invention, but R _{1 is a} non-halogen group.

In a fourth aspect of the invention, there is provided a compound, which is a compound of any one of compound 1 to compound 630.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

DRAWINGS

Figure 1 is a schematic representation of the concentration of neutrophil in compound 492 in a concentration-dependently inhibited urate sodium salt-induced peritonitis in mice.

Detailed ways

After long-term and intensive research, the present inventors have found that a compound having the structure of the formula (I) has an effect of inhibiting the activity of tubulin, and can be used for preparing or treating breast milk associated with abnormality of tubulin regulation. A drug for animal diseases. Based on the above findings, the inventors completed the present invention.

the term

As used herein, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of halogen, amino, hydroxy, nitro, cyano, trifluoromethyl. a C _1-12 alkyl or cycloalkyl group, a C _1-12 alkoxy group, an oxygen atom (ie, =0), a C _1-12 alkylamino group unsubstituted or substituted with a C _1-4 alkylamino group, C _2-6 ester group, C _2-6 acyl group, C _2-6 amide group, thio C _1-12 alkyl group, carboxyl group, C _5-12 aryl or heteroaryl group, C _5-12 heterocyclic group ( Containing from 1 to 5, preferably from 1 to 3, heteroatoms selected from N, O or S).

The term "C _1-12 alkyl" refers to a straight or branched alkyl group having from 1 to 12 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl. , tert-butyl, or the like.

The term "C _1-12 cycloalkyl" refers to a cycloalkyl group having from 1 to 12, preferably from 3 to 12 (ie C _3-12 ) carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, Cycloheptyl, or a similar group.

The term "C _1-12 alkoxy" refers to a straight or branched alkoxy group having from 1 to 12 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, Isobutoxy, sec-butoxy, tert-butoxy, or the like.

The term "halogen" refers to F, Cl, Br and I.

The term "C _1-12 alkylamino" refers to a C _1-12 alkyl group substituted by an amine group, for example having "C _1-12 alkyl-NH-" or "(alkyl) ₂ -N- (total number of carbon atoms) Is 1-12)", "-C _1-12 alkylene-NH ₂ ", "alkyl-N-alkylene- (total number of carbon atoms 1-12)", or "(alkyl) ₂ - a group of an N-alkylene-(having a total of 1-12 carbon atoms) structure, such as CH ₃ NH-, C ₂ H ₅ NH-, C ₃ H ₇ NH-, (CH ₃ ) ₂ N-, - CH ₂ NH ₂ , -C ₂ H ₅ NH ₂ , -C ₃ H ₇ NH ₂ , -C ₂ H ₄ N(CH ₃ ) ₂ , or the like. Wherein, the definition of C _1-12 alkyl is as defined above.

The term "C ₂ -C ₆ ester group" refers to a substitution of a structure such as "linear or branched alkyl/cycloalkyl/aryl/heteroaryl-carbonyl-oxy-- having 1 to 5 carbon atoms". A group such as an ethyl ester group, a propyl ester group, a butyl ester group, or the like.

The term "C ₁ -C ₆ amido" refers to a substitution of a structure such as "linear or branched alkyl/cycloalkyl/aryl/heteroaryl-carbonyl-amino-" having 0 to 5 carbon atoms. A group such as an acetamide group, a propionamide group, a butanamide group, or the like.

The term "C ₁ -C ₁₂ aryl" refers to an aryl group having 1 to 12 (preferably 6 to 10, ie C _6-10 ) carbon atoms, such as phenyl, naphthyl, etc., said aryl group being Is replaced or unsubstituted.

The term "C ₁ -C ₁₂ heteroaryl" refers to a heteroaryl group having 1 to 12 carbon atoms and one or more (preferably 1 to 3) heteroatoms selected from O, S and/or N, preferably 5 -8-membered heteroaryl. The heteroaryl group can be substituted or unsubstituted.

The term "C ₁ -C ₁₂ heterocyclyl" refers to a non-aromatic cyclic group having from 1 to 12 carbon atoms and one or more (preferably one to three) heteroatoms selected from O, S and/or N. The group is preferably a 5- to 8-membered heterocyclic group. The heterocyclic group may be substituted or unsubstituted.

In the present invention, the term "pharmaceutically acceptable" ingredient means a substance which is suitable for use in humans and/or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reaction), that is, a reasonable benefit/risk ratio.

In the present invention, the term "effective amount" means an amount of a therapeutic agent that treats, alleviates or prevents a target disease or condition, or an amount that exhibits a detectable therapeutic or prophylactic effect. The precise effective amount for a subject will depend on the size and health of the subject, the nature and extent of the condition, and the combination of therapeutic and/or therapeutic agents selected for administration. Therefore, it is useless to specify an accurate effective amount in advance. However, for a given condition, routine experimentation can be used to determine the effective amount that the clinician can determine.

Unless otherwise stated, all compounds appearing in the present invention are meant to include all possible optical isomers, such as a single chiral compound, or a mixture of various chiral compounds (i.e., racemates). Among all the compounds of the present invention, each of the chiral carbon atoms may be optionally in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

The term "compound of the invention" as used herein refers to a compound of formula I. The term also encompasses various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula I.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of the compound of the invention formed with an acid or base suitable for use as a medicament. Pharmaceutically acceptable salts include inorganic and organic salts. A preferred class of salts are the salts of the compounds of the invention with acids. Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, Organic acids such as maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzoic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

Preparation of compounds of formula (I)

The present invention provides a compound of the formula:

Wherein X ₁ , X ₂ , X ₃ , R ₁ , Ar ₁ , Ar ₂ , m and n are as defined above.

A preferred compound is any one of compounds 1-630.

The compound can be prepared by the following preparation method:

The method comprises the steps of:

(i-a) reacting a compound of formula a with a compound of formula b in an inert solvent to provide a compound of formula c;

(ii-a) reacting a compound of formula c with a compound of formula d in an inert solvent to provide a compound of formula e;

And when Q = NR ₁ , further comprising the optional step of: (iii-a) deprotecting a compound of formula e in an inert solvent to provide a compound of formula f;

Wherein X is a halogen; U is selected from the group consisting of halogen, OMs, OTs, boric acid, and pinacol borate; Q is O, S or NR ₁ ; P is selected from the group consisting of p-methoxybenzyl, benzyl. , tert-butoxycarbonyl;

Or the method comprises the steps of:

(i-b) in an inert solvent, the compound of formula g is substituted with a halogen to give a compound of formula h;

Wherein R ₁ is halogen (fluorine, chlorine, bromine or iodine).

The compounds can also be prepared by the following steps:

(i-c) a compound of formula a is subjected to a ring closure reaction with a compound of formula b in an inert solvent to provide a compound of formula c;

Preferably, the ring closure reaction is carried out by inorganic salt catalysis;

Wherein V is halogen, OMs, or OTs; the remaining groups are as defined above, but R _{1 is a} non-halogen group.

In the above synthesis reaction, the respective inert solvents are not particularly limited and may be selected from the group consisting of dichloromethane, chloroform, carbon tetrachloride, tetrahydrofuran (THF), acetonitrile, dimethylformamide (DMF), and Methyl sulfoxide (DMSO), ethylene glycol dimethyl ether, 1,2-dichloroethane, dimethyl phthalate (DMP), N-methylpyrrolidone (NMP), methanol, ethanol, n-butyl A solvent for alcohol, isopropanol, petroleum ether, ethyl acetate, n-hexane or diethyl ether.

In the above preparation process, the necessary base may be selected from, for example, sodium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, sodium hydrogencarbonate, calcium carbonate, potassium phosphate, sodium methoxide, sodium ethoxide, sodium t-butoxide, tert-butanol. Potassium, diethylamine, triethylamine, N,N-diisopropylethylamine (DIPEA) or 1,8-diazabicyclo [5.4.0]undec-7-ene (DBU); The acid may be selected from trifluoroacetic acid, hydrochloric acid, sulfuric acid, nitric acid, methanesulfonic acid or p-toluenesulfonic acid and the like.

In the above preparation process, the necessary inorganic salts may be selected from sodium fluoride (NaF), potassium fluoride (KF), cesium chloride, aluminum chloride and the like.

A catalyst may optionally be employed in each step of the above preparation process, and the catalyst may be selected from the group consisting of tetrakistriphenylphosphorus palladium (Pd(PPh ₃ ) ₄ ), palladium acetate (Pd(OAC) ₂ ), dichloro Palladium (PdCl ₂ ) palladium carbon, ditriphenylphosphine palladium dichloride (PdCl ₂ (PPh) ₂ ), 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (PdCl ₂ (dppf) ₂ ), tris(dibenzylideneacetone)dipalladium (Pd ₂ (dba) ₃ ), iodine (or bromine or chlorine) cuprous, iodine (or bromine or chlorine) copper, copper powder, etc. . When it is desired to use a catalytic ligand, the necessary catalytic ligand may be selected from the group consisting of triphenylphosphine, 2-biscyclohexylphosphine-2', 4',6'-triisopropylbiphenyl (Xphos) and 2-dicyclohexyl. Phosphine-2',6'-dimethoxybiphenyl (Sphos) and the like.

The organometallic reagent may be n-butyllithium, sodium borohydride, lithium borohydride or sodium triacetylborohydride or the like. The halogenating agent may be phosphorus tribromide, phosphorus oxychloride, bromine, iodine, nitrogen chloride (or bromine or iodine) succinimide (NCS, NBS or NIS) or phenyl trimethyl three. Ammonium bromide.

The oxidizing agent may be a Dess-Martin oxidizing agent, a Swern oxidizing agent, m-chloroperoxybenzoic acid, chlorodichromic acid pyridine (PDC) or chlorochromic acid pyridine (PCC).

Pharmaceutical composition and method of administration

Since the compound of the present invention has excellent inhibitory activity against tubulin, the compound of the present invention and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the present invention are mainly The pharmaceutical composition of the active ingredient can be used for the treatment, prevention, and alleviation of diseases associated with tubulin activity or expression levels, particularly for diseases in which tubulin activity or expression levels are all related. According to the prior art, the compounds of the invention are useful in the treatment of diseases such as cancer, neurodegenerative diseases, malaria, AIDS, gout, diabetes and the like.

The pharmaceutical compositions of the present invention comprise a safe or effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. By "safe and effective amount" it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 1 to 2000 mg of the compound of the invention per agent, more preferably from 5 to 200 mg of the compound of the invention per agent. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By "compatibility" it is meant herein that the components of the composition are capable of intermingling with the compounds of the invention and with each other without significantly reducing the efficacy of the compound. Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid). , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as

), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration. .

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, hard Calcium citrate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.

In addition to these inert diluents, the compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening Agents, flavors and fragrances.

In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.

Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants. The active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.

The compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight, The dose to be administered is usually from 1 to 2000 mg, preferably from 5 to 500 mg. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.

The main advantages of the present invention over the prior art include:

(1) A novel structure of a compound having a tubulin inhibitory activity is provided.

(2) Providing a compound having tumor suppressing activity, which compound can be used for the preparation of a medicament for treating a tumor.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

The nuclear magnetic resonance spectrum was recorded using a Bruker AMX-400 type, a Gemini-300 type or an AMX-600 type nuclear magnetic resonance apparatus, and the unit of the chemical shift δ was ppm. All solvents are analytically pure reagents. Color development was carried out by iodine or ultraviolet fluorescence. The organic solvent was distilled off under reduced pressure in a rotary evaporator. The starting reactants used in the present invention are commercially available unless otherwise specified.

It should be noted that, in the following examples, the conventional post-treatment method is: after the reaction is completed, an appropriate amount of an organic solvent and water are added to the reaction liquid, the organic phase and the aqueous phase are separated, the organic phase is combined, and dried using NaSO ₄ . After filtration, it was evaporated to dryness under reduced pressure to give a crude material, which was purified by column chromatography to give the final product.

Preparation examples:

Example 1: Preparation of Compound 1 (refer to the scheme shown in the following reaction formula)

Step (i): N-PMB-5-iodo-3-chloropyrrole (1 g, 1 eq), p-methoxyaniline (1.06 g, 3 eq) and potassium phosphate (1.22 g, 2 eq) were dissolved in 1,4-di In 30 mL of hexacyclohexane solution, nitrogen was added, copper iodide (55 mg, 10%) was added, and the reaction was completed at 110 °C for 24 hours. The solvent was directly evaporated to give N-PMB-5-(4-methoxyl). Aniline)-3-chloropyrrole (532 mg);

Step (ii): N-PMB-5-(4-methoxyaniline)-3-chloropyrrole (500 mg, 1 eq), 2-thiopheneboronic acid (280 mg, 1.5 eq) Potassium phosphate (619 mg, 2 eq) was dissolved in 5 mL of n-butanol, protected with nitrogen, and palladium acetate (16 mg, 0.05 eq) and xphos (70 mg, 0.1 eq) were added and reacted at 110 degrees for 12 hours. Directly evaporating the solvent column to obtain N-PMB-5-(4-methoxyanilino)-3-(2-thienyl)pyrrole (319 mg);

Step (iii): N-PMB-5-(4-methoxyanilino)-3-(2-thienyl)pyrrole (100 mg) was dissolved in 20 mL of trifluoroacetic acid and stirred at room temperature overnight. The saturated sodium hydrogencarbonate was adjusted to a weakly basic pH, extracted with ethyl acetate, and the organic layer was dried and evaporated to dryness to afford compound 1[5-(4-methoxyanilinyl)-3-(2-thienyl)pyrrole] (47mg);

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.91 (s, 1H), 7.70 (dd, J = 8.5,3.1Hz, 1H), 7.58-7.48 (m, 2H), 7.40 (dd, J = 8.3,3.1 Hz, 1H), 7.13 (t, J = 7.9 Hz, 1H), 7.06 - 6.94 (m, 2H), 6.61 (d, J = 7.9 Hz, 1H), 5.89 (d, J = 3.1 Hz, 1H), 3.79(s,3H).

The compound of Example 2-55 was prepared in the same manner as in Example 1 except that the compound b or d was different.

Preparation of Compound 60 of Example 60

Step (i): 5-bromo-3-chlorofuran (1 g, 1 eq), p-methoxyaniline (2.04 g, 3 eq) and potassium phosphate (2.34 g, 2 eq) were dissolved in 15 mL of N-methylpyrrolidone, nitrogen Protection, adding cuprous bromide (158 mg, 20%), reacting at 150 °C for 24 hours, extracting with ethyl acetate, drying the organic layer and drying the column to give 5-(4-methoxyaniline)-3-chloro Furan (613mg);

Step (ii): 5-(4-methoxyaniline)-3-chlorofuran (500 mg, 1 eq), 2-methylimidazolium [1,2-a]pyrimidine-3-boronic acid (791 mg, 2 eq) The cesium fluoride (679 mg, 2 eq) was dissolved in 15 mL of tetrahydrofuran, and protected with nitrogen. Pd2(dba)3 (205 mg, 0.1 eq) and tri-tert-butylphosphonate (90 mg, 0.2 eq) were added, and the reaction was refluxed for 10 hours, and the reaction was completed. Directly evaporating the solvent column to give 5-(4-methoxyanilino)-3-(2-methylimidazolium [1,2-a]pyrimidin-3-)furan (107 mg);

^{1 H NMR (400MHz, CDCl3)} δ9.58 (dd, J = 7.9,3.0Hz, 1H), 8.77 (dd, J = 8.0,3.0Hz, 1H), 7.78-7.43 (m, 2H), 7.27 (t , J = 8.2 Hz, 1H), 7.03 (ddd, J = 8.2, 6.7, 3.3 Hz, 3H), 5.93 (d, J = 7.0 Hz, 1H), 3.79 (s, 3H), 2.61 (s, 3H) .

The preparation methods of Examples 56-59 and 61-238 are the same as those of Example 60 except that the compounds b or d are different.

Preparation of Compound 501 of Example 501 (refer to the scheme shown in the following reaction formula)

0.1 g (1 eq) of p-methoxythiourea and 0.215 g (1 eq) of a heterocyclic compound were added to a 25 ml three-necked flask, and 6 mL of methanol and water (volume: volume = 1:1) were added thereto, and catalysis was added under nitrogen protection. The amount of NaF (2.3 mg, 0.1 eq) was reacted at room temperature for 1-5 minutes. The reaction was completed by TLC, washed with saturated sodium bicarbonate solution and extracted with dichloromethane (10 mL×3). .

^{1 H NMR (400MHz, CDCl3)} δ8.95 (d, J = 7.9Hz, 1H), 7.90 (d, J = 8.1Hz, 1H), 7.55 (d, J = 8.0Hz, 2H), 7.11 (s, 1H), 7.02 (d, J = 7.8 Hz, 2H), 3.79 (s, 3H), 2.61 (s, 3H), 2.36 (s, 3H).

The preparation methods of Examples 239-500, 502-618 and Examples 624-630 were the same as those of Example 501 except that the compounds a or b were different, as shown in the following table.

Preparation of Example 619 Compound 619

The compound (100 mg, 1 eq) was dissolved in 2 mL of acetic acid, and nitrobromosuccinimide (54 mg, 1.5 eq) was added thereto, and the reaction was carried out at 60 °C for 12 hours, and the reaction was completed. The mixture was extracted with EtOAc EtOAc (EtOAc m. ^{1 H NMR (400MHz, CDCl3)} δ8.55 (dd, J = 15.0,2.9Hz, 1H), 7.61-7.48 (m, 2H), 7.40 (dd, J = 14.9,3.0Hz, 1H), 7.08-6.93 (m, 3H), 3.77 (s, 3H), 2.59 (s, 3H).

The preparation of Examples 620-621 was the same as Example 619 except that Compound a was different, see the table below.

Preparation of Example 622 Compound 622

The compound (200 mg, 1 eq) was dissolved in acetonitrile / water (4 mL / 1 mL), and then trifluoroiodomethane (157.6 mg, 1.5 eq), sodium sulphate (93.4 mg, 1 eq) and sodium bicarbonate (45 mg) ), reacted at room temperature for 12 hours, and the reaction was completed. After adding 20 mL of water and ethyl acetate (10 mL×3), the organic layer was dried over sodium sulfate and evaporated to dryness. ^{1 H NMR (400MHz, CDCl3)} δ8.48 (dd, J = 7.9,3.0Hz, 1H), 8.39 (s, 2H), 7.33 (dd, J = 7.9,3.0Hz, 1H), 6.96 (t, J = 8.0 Hz, 1H), 3.76 (s, 3H), 2.52 (s, 3H).

Preparation of Example 623 Compound 623

The compound (100 mg, 1 eq) was dissolved in anhydrous acetonitrile (5 mL), EtOAc (br. 10 mL of water was quenched, ethyl acetate (10 mL×3) was evaporated. ^{1 H NMR (400MHz, CDCl3)} δ8.56 (d, J = 8.3Hz, 1H), 8.47 (s, 2H), 7.41 (d, J = 9.0Hz, 1H), 7.04 (t, J = 7.9Hz, 1H), 3.84 (s, 3H), 2.59 (s, 3H).

Compound in vitro bioactivity test

Determination of biochemical activity of microtubule-associated protein

Principle of measurement: Microtubules are multimers of tubulin. Tubulin α and β are joined end to end to form a heterodimer, which is then multi-polymerized into microtubule fibrils. The microtubules are composed of 13 fibrils, and each micrometer long microtubule is composed of 1,650 heterodimers. In vitro, tubulin can be polymerized. The effect of the compound on tubulin polymerization was examined using a Tubulin polymerization assay kit (BK011P, Cytoskeleton, Inc.). The kit contains specific reporter fluorescence, which is inserted into the microtube during microtubule polymerization, and the polymerization of the microtube can be monitored according to the intensity of the fluorescence. First, add 5 μl of the above compound or control compound at different concentrations in a 96-blackboard, preheat at 37 ° C, and add 45 μl of microtubule polymerization mixture (243 μl Buffer1, 112 μl Tubulin Glycerol Buffer, 4.4 μl GTP stock, 85 μl Tubulin) to each well. Stock), the fluorescence intensity was measured at a wavelength of Ex.=360 nm and Em.=450 nm per minute at 37 ° C in a microplate reader, and the polymerization of the microtubule was monitored for one hour to calculate the IC50 of the compound inhibiting tubulin polymerization. The test results are shown in Table 1. In Table 1, microtubule polymerization inhibitory activity (IC ₅₀ ) indicates a method: + represents 1-10 μM; ++ represents 0.1-1 μM; and +++ represents < 0.1 μM.

Table 1 results of inhibition of microtubule polymerization activity test

Test results and analysis:

The IC ₅₀ in the above table refers to the inhibitor concentration (50% inhibitory concentration) when the microtubule polymerization is inhibited by half.

As can be seen from the results in the above table, the above compounds also significantly inhibited the polymerization of tubulin as compared with the positive control vincristine (VCR).

In vitro antitumor activity screening of compounds

Principle of the assay: Colon cancer cells HCT116 were cultured in modified 5A medium containing 10% fetal bovine serum and trypsinized for passage. After about 70% of cell fusion, trypsinization was performed to prepare a cell suspension, and the cells were counted under a microscope, and then seeded in a 96-well plate at 5 × 10 ³ cells per well. After the overnight culture, the above compound was administered. Changes in cell proliferation after 72 hours of administration were observed by MTT method. After the compound was treated for 72 hours, 10 μl of MTT working solution (5 mg/ml) was added to each well, and the mixture was incubated at 37 ° C for 2 hours, and then the culture solution was aspirated, and 100 μl of DMSO was added to each well. After shaking for 15 minutes at room temperature, the OD value was measured at a wavelength of 492 nm by a microplate reader. The activity of the compound for inhibiting the proliferation of HCT116 cells was calculated. The specific inhibition rates are shown in Tables 2 and 3, wherein the inhibition of cell proliferation activity (IC ₅₀ ) indicates the method: + means 1-10 μM; ++ means 0.1-1 μM; +++ means 0.01-0.1 μM; ++++ means 0.01-0.001 μM; +++++ means <0.001 μM; "-" means no activity. IC ₅₀ represents the concentration of 50% growth inhibition required for cell growth inhibition.

Cell viability (%) is calculated as:

Survival rate (%) = (administered well OD - blank well OD) / (control well OD - blank well OD) × 100

Inhibition rate (%) = 1 - survival rate (%)

Other cancer cell lines used are Hela (human cervical cancer cells), MCF-7 (human breast cancer cells), LM3 (human liver cancer cells), NCI-N87 (human gastric cancer cells), and Caki-1 (human kidney cancer cells). ), A549 (human lung cancer cells), HT1080 (human fibrosarcoma cells), A431 (human epidermal squamous cell carcinoma cells), PC3 (human prostate cancer cells), HL60 (human leukemia cells), Panc-1 (human pancreatic cancer) Cells), KB (human oral cancer cells), U87-MG (human glioma cells), K562 (human chronic granulocyte leukemia cells), Kasumi-1 (human leukemia cells), THP-1 (human leukemia cells) ), Jurkat (human T lymphocytic leukemia cells), REH (human B lymphocyte leukemia cells), Raji (human Burkitt cell leukemia cells), RNK-16 (human NK cell leukemia cells), KMS-1 (human multiple bone marrow) Tumor cells), P39 (human myelodysplastic syndrome cells), U118-MG (human glioblastoma cells), H4 (human glioma cells), SK-N-SH (human neuroblastoma cells) , SH-SY5Y (human neuroblastoma cells), A549/Taxol (Paclitaxel-resistant human lung cancer cells), KB/VCR (Vincristine-resistant oral cancer cells), K562/Adr (Anomycin) Human chronic myeloid leukemia cells), respectively, with DMEM + 10% FBS culture medium or use the 1640 + 10% FBS culture.

Table 2 Compounds inhibit the growth of cancer cells, broad-spectrum biological activity test results

As can be seen from the above experimental results, the compounds of the present invention have inhibitory activities against the growth of different types of tumor cells, suggesting that the compounds of the present invention have a broad spectrum of antitumor activity.

Table 3 Activity test results of some compounds inhibiting blood cancer cell growth

Numbering	Kasumi-1	THP-1	Jurkat	REH	Raji	RNK-16	KMS-1	P39
VCR	++++	++++	++++	++++	++++	++++	++++	++++
Taxol	++++	++++	++++	++++	++++	++++	++++	++++
149	++++	++++	+++	+++	+++	+++	+++	+++
153	++++	+++	+++	+++	+++	+++	+++	+++
167	++++	+++	++++	+++	++++	+++	++++	+++
253	+++	+++	+++	+++	+++	+++	+++	+++
257	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
272	++++	++++	++++	++++	++++	++++	++++	++++
275	++++	++++	++++	++++	++++	++++	++++	++++
285	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
305	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
325	+++++	+++	+++++	+++	+++++	+++	+++++	+++++
359	++++	++++	++++	++++	++++	++++	++++	++++
361	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
370	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
374	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
387	+++	+++++	+++	+++++	+++	+++++	+++	+++++
392	+++++	+++	+++	+++++	+++	+++++	+++	+++++
413	+++	+++++	+++	+++	+++	+++	+++	+++
414	++++	++++	++++	++++	++++	++++	++++	++++
417	++++	++++	++++	++++	++++	++++	++++	++++
433	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
436	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
492	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
493	++++	++++	++++	++++	++++	++++	++++	++++
499	++++	++++	++++	++++	++++	++++	++++	++++
501	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
505	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
519	++++	++++	++++	++++	++++	++++	++++	++++
520	++++	++++	++++	++++	++++	++++	++++	++++
526	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
527	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
534	++++	++++	++++	++++	++++	++++	++++	++++
536	++++	++++	++++	++++	++++	++++	++++	++++
540	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
556	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
557	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
561	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
565	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
583	++++	++++	++++	++++	++++	++++	++++	++++
585	++++	++++	++++	++++	++++	++++	++++	++++
586	++++	++++	++++	++++	++++	++++	++++	++++
589	++++	++++	++++	++++	++++	++++	++++	++++
590	++++	++++	++++	++++	++++	++++	++++	++++
591	++++	++++	++++	++++	++++	++++	++++	++++
609	+++	+++	+++	+++	+++	+++	+++	+++
617	++++	++++	++++	++++	++++	++++	++++	++++

619	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
620	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++
621	+++++	+++++	+++++	+++++	+++++	+++++	+++++	+++++

It can be seen from the above experimental results that the compounds of the present invention are useful for various types of blood cancer cells (acute myeloid leukemia cells, acute or chronic lymphocytic leukemia cells, multiple myeloma cells, myelodysplastic syndrome cells). The growth of both has an inhibitory activity, suggesting that the compounds of the present invention have a broad spectrum of activity for inhibiting the growth of blood cancer cells.

Table 4 Activity test results of some compounds inhibiting the growth of brain cancer cells

It can be seen from the above experimental results that the compound of the present invention has an inhibitory activity on the growth of brain tumor cells (glioma cells and neuroblastoma cells), suggesting that the compound of the present invention is suitable for inhibiting the growth of brain tumor cells. active.

Table 5 results of some compounds against growth inhibition test of drug-resistant cancer cells

	KB	KB/VCR	A549	A549/Taxol	K562	K562/Adr
VCR	++++	+	++	++
Taxol	++++	++	+++	+
Adr					+++	+
149	++++	++	++	++	+++	+++
153	++++	++	+++	++	+++	+++
167	++++	++	+++	++	+++	+++
253	+++	++	++	++	+++	+++
257	++++	++++	++++	++++	+++++	+++++
272	+++	+++	+++	+++	++++	++++
275	+++	+++	+++	+++	++++	++++
285	++++	++++	++++	++++	+++++	+++++
305	++++	++++	++++	++++	+++++	+++++
325	++	++	++	++	+++	+++
359	+++	+++	+++	+++	++++	++++
361	++++	++++	++++	++++	+++++	+++++
370	++++	++++	++++	++++	+++++	+++++
374	++++	++++	++++	++++	+++++	+++++
387	++	++	++	++	+++	+++
392	++	++	++	++	+++	+++
413	++	++	++	++	+++	+++
414	+++	+++	+++	+++	++++	++++

417	+++	+++	+++	+++	++++	++++
433	++++	++++	++++	++++	+++++	+++++
436	++++	++++	++++	++++	+++++	+++++
492	++++	++++	++++	++++	+++++	+++++
493	+++	+++	+++	+++	++++	++++
499	+++	+++	+++	+++	++++	++++
501	++++	++++	++++	++++	+++++	+++++
505	++++	++++	++++	++++	+++++	+++++
519	+++	+++	+++	+++	++++	++++
520	+++	+++	+++	+++	++++	++++
526	++++	++++	++++	++++	+++++	+++++
527	++++	++++	++++	++++	+++++	+++++
534	+++	+++	+++	+++	++++	++++
536	+++	+++	+++	+++	++++	++++
540	++++	++++	++++	++++	+++++	+++++
556	++++	++++	++++	++++	+++++	+++++
557	++++	++++	++++	++++	+++++	+++++
561	++++	++++	++++	++++	+++++	+++++
565	++++	++++	++++	++++	+++++	+++++
583	+++	+++	+++	+++	++++	++++
585	+++	+++	+++	+++	++++	++++
586	+++	+++	+++	+++	++++	++++
589	+++	+++	+++	+++	++++	++++
590	+++	+++	+++	+++	++++	++++
591	+++	+++	+++	+++	++++	++++
609	++	++	++	++	+++	+++
617	+++	+++	+++	+++	++++	++++
619	++++	++++	++++	++++	+++++	+++++
620	++++	++++	++++	++++	+++++	+++++
621	++++	++++	++++	++++	+++++	+++++

As can be seen from the results in the above table, the above compounds have significant inhibition of the listed cancer cells compared with the positive controls of vincristine (VCR), paclitaxel (Taxol) and adriamycin (Adrmycin, Adr). The activity of growth of drug-resistant cancer cells, especially for paclitaxel, vincristine or doxorubicin-resistant strains, has a good inhibitory effect on the growth of resistant strains of chronic myeloid leukemia cells.

Compound 492 inhibits neutrophils to relieve gout symptoms

Tubulin inhibitors are widely used, in addition to their use as anti-neoplastic agents, in the treatment of gout, as well as antifungal agents and broad-spectrum anthelmintics.

Experimental content: 1 mg of sodium urate was dissolved in 0.5 ml of endotoxin-free phosphate buffer solution to prepare a solution. C57BL/6 mice were intraperitoneally injected with sodium urate solution to establish a model of peritonitis. On days 1-4 after injection of sodium urate solution, mice were treated with different concentrations of compound 492 (0, 0.25, 0.5, and 1 mg/kg) daily, and compound 492 was found to significantly inhibit the gout model in mice. The neutrophil flow (Figure 1) inhibits inflammation and relieves gout in mice.

As can be seen from Fig. 1, the compound 492 inhibited the neutrophil flow in the peritonitis mice induced by sodium urate salt crystal in a concentration-dependent manner, suggesting that the compound exhibited an effect of relieving the ventilation condition in the mouse in vivo experiment.

It should be noted that the above-described embodiments are only intended to illustrate and not to limit the technical solutions of the present invention, and any equivalent substitutions or modifications are considered to be included in the scope of the present invention.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (10)

1. Use of a compound of formula (I), an isomer, a racemate, a pharmaceutically acceptable salt, a crystalline hydrate, a solvate or a mixture thereof, for use in (a) preparation of a treatment Or drugs that prevent mammalian diseases associated with abnormal expression of tubulin; (b) inhibit microtubule polymerization activity; (c) inhibit tumor cell growth:

   

   among them:

   X _{1 is} selected from N or CR ₁ (when R1 is H, denoted as C);

   X _{2 is} selected from NR ₁ , O or S;

   X _{3 is} selected from the group consisting of NH, O, S, C=O, C=S, C=NH, -(C=O)-NH-, -(C=O)-O-,-(C=O )-S-,-(C=S)-NH-,-(C=S)-O-,-(C=S)-S-,-(C=NH)-NH-,-(C=NH )-O- or -(C=NH)-S-;

   R _{1 is} selected from the group consisting of hydrogen, hydrazine, halogen, amino, hydroxy, nitro, cyano, carboxy, C _2-6 ester, C _1-6 amide, unsubstituted or halogenated C _1-12 alkane Or cycloalkyl, -CH ₂ -Y-(C _1-12 alkyl or cycloalkyl) (wherein Y is O or NH or S), C _1-12 aryl or heteroaryl, -CH ₂ - (C _1-12 aryl or heteroaryl);

   The Ar ₁ group is a substituted or unsubstituted C _1-18 aryl or heteroaryl group, a substituted or unsubstituted -CH ₂ -(C _1-12 aryl or heteroaryl), substituted or unsubstituted C _{1 -18} heterocyclic group, substituted or unsubstituted -CH ₂ -(C _1-12 heterocyclic group);

   The Ar ₂ group is hydrogen, substituted or unsubstituted C _1-12 alkyl or cycloalkyl, substituted or unsubstituted -CH ₂ -Y-(C _1-12 alkyl or cycloalkyl) (where Y is O or NH or S), substituted or unsubstituted C _1-12 aryl or heteroaryl, substituted or unsubstituted -CH ₂ -(C _1-12 aryl or heteroaryl), substituted or unsubstituted C _1-12 heterocyclic group;

   m is selected from 0, 1, 2, 3, 4, 5, 6;

   n is selected from 0, 1, 2, 3, 4, 5, 6;

   Wherein the substitution is substituted with one or more substituents selected from the group consisting of halogen, amino, hydroxy, nitro, cyano, trifluoromethyl, C _1-12 alkyl or cycloalkyl, a C _1-12 alkoxy group, an oxygen atom (ie, =0), a C _1-12 alkylamino group unsubstituted or substituted with a C _1-4 alkylamino group, a C _2-6 ester group, a C _2-6 acyl group, C _1-6 amido, thio C _1-12 alkyl group, a substituted carboxyl group, unsubstituted or substituted with 1-5 halogen, amino, hydroxy, nitro, cyano, trifluoromethyl C _1-12 aryl group Or a heteroaryl group, or a C _1-12 heterocyclic group which is unsubstituted or substituted by 1 to 5 halogen, amino, hydroxy, nitro, cyano, trifluoromethyl (containing 1-5, preferably 1-3 a hetero atom selected from N, O or S).
2. The use of claim 1 wherein:

   The X _{3 is} selected from the group consisting of NH, O, S, C=O, C=S, C=NH, -(C=O)-NH-, -(C=O)-O-,-(C =O)-S-,-(C=S)-NH-,-(C=S)-O-,-(C=S)-S-;

   The R _{1 is} selected from the group consisting of hydrogen, hydrazine, halogen, amino, hydroxy, nitro, cyano, carboxy, ester, amide, unsubstituted C _1-6 alkyl or cycloalkyl, (1 3) fluoro C _1-6 alkyl or cycloalkyl, (1-3) C _1-6 amine substituted C _1-6 alkyl or cycloalkyl, (1-3) C _1-6 alkoxy Substituted C _1-6 alkyl or cycloalkyl, -CH ₂ -Y-(C _1-6 alkyl), C _1-12 aryl or heteroaryl, -CH ₂ -(C _1-12 aryl or Heteroaryl).
3. The use according to claim 1, wherein in the Ar ₁ or Ar ₂ group, the C _5-12 aryl or heteroaryl group is selected from the group consisting of:

   

   m is 0, 1, 2, 3, 4, 5;

   n is 0, 1, 2, 3, 4, 5.
4. The use according to claim 1, wherein said Ar _{1 is} selected from the group consisting of C _1-8 heteroaryl and C _2-10 heteroaryl, C _4-8 heteroaryl and C _{2- 10} heterocyclic group.
5. The use according to claim 1, wherein the compound is a compound 1 to a compound 630 selected from the following table (where a thick horizontal line

   

   Mark the connection part):

   Table 1

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   

   
6. The use of the pharmaceutical composition according to claim 1, wherein the mammalian disease associated with abnormal regulation of microtubule-associated proteins is a disease selected from the group consisting of cancer, neurodegenerative diseases, malaria, AIDS, Gout, diabetes; preferably selected from the group consisting of lymphoma, lung cancer, gastric cancer, pancreatic cancer, breast cancer, prostate cancer, leukemia, brain tumor, and cervical cancer.
7. The use according to claim 1, wherein the tumor cells are tumor cells selected from the group consisting of colon cancer cells, cervical cancer cells, breast cancer cells, liver cancer cells, gastric cancer cells, renal cancer cells, and lung cancer. Cells, fibrosarcoma cells, epidermal squamous cell carcinoma cells, prostate cancer cells, leukemia cells, pancreatic cancer cells, oral cancer cells, glioblastoma cells, neuroblastoma cells, paclitaxel-resistant lung cancer cells, vincristine-resistant oral cavity Cancer cells, resistant to doxorubicin chronic granulocyte leukemia cells.
8. A method for preparing a compound represented by the formula (I):

   

   Wherein X ₁ , X ₂ , X ₃ , R ₁ , Ar ₁ , Ar ₂ , m and n are as defined in any of claims 1-5;

   The method comprises the steps of:

   

   (i-a) reacting a compound of formula a with a compound of formula b in an inert solvent to provide a compound of formula c;

   (ii-a) reacting a compound of formula c with a compound of formula d in an inert solvent to provide a compound of formula e;

   And when Q = NR ₁ , further comprising the optional step of: (iii-a) deprotecting a compound of formula e in an inert solvent to provide a compound of formula f;

   Wherein X is a halogen; U is selected from the group consisting of halogen, OMs, OTs, boric acid, and pinacol borate; Q is O, S or NR ₁ ; P is selected from the group consisting of p-methoxybenzyl, benzyl. , tert-butoxycarbonyl;

   Or the method comprises the steps of:

   

   (i-b) in an inert solvent, the compound of formula g is substituted with a halogen to give a compound of formula h;

   Wherein R ₁ is halogen (fluorine, chlorine, bromine or iodine)

   The remaining groups are as defined in claim 1.
9. A method for preparing a compound represented by the formula (I):

   

   Wherein X ₁ , X ₂ , X ₃ , R ₁ , Ar ₁ , Ar ₂ , m and n are as defined in any of claims 1-5;

   The method comprises the steps of:

   

   (i-c) a compound of formula a is subjected to a ring closure reaction with a compound of formula b in an inert solvent to provide a compound of formula c;

   Preferably, the ring closure reaction is carried out by inorganic salt catalysis;

   Wherein V is halogen, OMs, or OTs; the remaining groups are as defined in claim 1, but R _{1 is a} non-halogen group.
10. A compound represented by the following formula (I), characterized in that the compound is a compound 1 to a compound 630 selected from the following table (wherein a bold line

    

    Mark the connection part):

    

    Table 1

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

    

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