CN101265250A

CN101265250A - Substituted flavonoids and preparation method, application and pharmaceutical composition thereof

Info

Publication number: CN101265250A
Application number: CNA2007100381573A
Authority: CN
Inventors: 柳红; 蒋华良; 朱维良; 郑鸣芳; 郑明月; 罗小民; 沈建华; 陈凯先
Original assignee: Shanghai Institute of Materia Medica of CAS
Current assignee: Shanghai Institute of Materia Medica of CAS
Priority date: 2007-03-16
Filing date: 2007-03-16
Publication date: 2008-09-17

Abstract

The invention provides a substituted flavonoid in formula (I), and pharmaceutically acceptable salt, solvate and hydrate thereof, wherein the definitions of R1-R10, X and Y are described in specification. Pharmacological experiment indicates that the compound is 5-lipoxidase strong inhibitor. Therefore, the flavonoid is expected to be applied in the preparation of drugs for preventing and/or treating inflammation diseases, immunological diseases such as asthma, anaphylaxis, etc., and cardiovascular disease. The invention also provides the preparation method of the flavonoid.

Description

Substituted flavonoid compound, preparation method, application and pharmaceutical composition thereof

Technical Field

The invention relates to the field of medicinal chemistry and pharmacotherapeutics, in particular to a substituted flavonoid compound, a preparation method thereof, application of the substituted flavonoid compound serving as a 5-lipoxygenase inhibitor in preparing medicaments for treating and/or preventing inflammatory diseases, immune diseases such as asthma, allergy and the like and cardiovascular diseases, and a pharmaceutical composition containing the compound.

Background

5-lipoxygenase (5-lipoxygenase, 5-LOX) is a member of the family of lipoxidases, widely present in tissues of animals and plants. Lipperoxidase catalyzes the oxidation of polyunsaturated fatty acids (e.g., arachidonic acid, AA) having 1, 4-cis-pentadiene segments to produce their peroxidation products. When AA is composed of phospholipid membrane A in human body₂After release, 5-LOX oxidizes the C-5 position to 5-hydroxyperoxytetraenic acid (5-HPETE) and subsequent dehydration to the primary intermediate LTA₄。LTA₄Has an epoxy structure, is very unstable in vivo, and is easy to form eicosenoic acid, namely Leukotrienes (LTs), through biotransformation. LTA₄In LTB₄LTB formation of dihydroxy acid under the action of synthetase₄；LTA₄At LTC₄The LTC is formed by combining with glutathione under the action of synthetase₄；LTC₄LTD formation after loss of a glutamic acid molecule by glutamyltransferase₄(ii) a And LTD₄And can lose a molecule of aminoacetic acid to form LTE under the action of related dipeptidase₄(see chemical reaction formula 1). LTC₄、LTD₄And LTE₄Also known as cysteinyl leukotrienes or peptide leukotrienes, these leukotrienes are mostly produced by inflammatory cells.

Chemical reaction formula 1

LTs have a wide range of biological activities and are regulated by specific G-protein coupled receptors. LTB₄There are strong inflammatory cells (e.g.:neutrophils, macrophages, eosinophils, etc.) chemotactic effects associated with the tendency of leukocytes to metastasize to sites of inflammation. By activation of neutrophils, LTB₄Can cause cell degranulation and release of related enzymes, with concomitant production of peroxides. LTB₄Can also enhance the adhesion of the leukotriene compound to vascular endothelial cells and promote tissue penetration. In addition, LTB₄Also plays an important role in immune responses, namely enhancing the release of proinflammatory cytokines from macrophages and lymphocytes.

Cysteinyl leukotrienes were originally considered as "allergenic slow-reacting substances" as biological mixtures. They play a pathophysiological role in perceptual hypersensitivity reactions. They are strong smooth muscle contractants (100-1000 times stronger than histamine), and are especially able to cause bronchoconstriction. They also promote mucus secretion and play an important role in bronchial smooth muscle cell proliferation. In the microcirculatory system, they enhance vascular permeability by contracting endothelial cells, thereby causing edema from plasma extravasation. In view of these potential biological activities, LTs have been recognized as important regulators of numerous inflammatory diseases and allergic reactions, such as rheumatoid arthritis, inflammatory bowel disease, ulcerative colitis, asthma, psoriasis, allergic rhinitis, and the like. Further, as studies on 5-LOX and AA metabolites have been conducted, it has been found that leukotriene compounds are closely related to cardiovascular diseases such as arteriosclerosis, myocardial infarction and stroke, cell proliferation and tumors. Thus, the development of 5-LOX inhibitors has been a field of great interest, and there are currently a large number of small molecule 5-LOX inhibitors reported, mainly in the three major classes redox, iron-chelating and non-redox. However, only Zileuton is one of the 5-LOX inhibitors actually marketed for clinical use, and thus, the market is urgently in need of the 5-LOX inhibitor having a remarkable effect and a novel structure.

The inventor obtains a class of substituted flavonoid compounds by researching natural product Artonin E analogues and modifying synthetic intermediates thereof, and the substituted flavonoid compounds have good inhibitory activity on 5-LOX at a cellular level.

Disclosure of Invention

The invention aims to provide a substituted flavonoid compound with 5-LOX potent inhibitory activity and pharmaceutically acceptable inorganic or organic salts, solvates or hydrates thereof.

The invention also aims to provide a preparation method of the compound.

Still another object of the present invention is to provide a pharmaceutical composition for preventing and/or treating inflammatory diseases, immunological diseases such as asthma and allergy, and cardiovascular diseases, comprising a substituted flavonoid compound and a pharmaceutically acceptable salt thereof or a solvate or hydrate thereof.

The invention also aims to provide application of the substituted flavonoid compound and the pharmaceutically acceptable salt thereof or the solvate or hydrate thereof as a 5-LOX inhibitor in preparing medicines for preventing and/or treating immune diseases such as inflammatory diseases, asthma, allergy and the like and cardiovascular diseases.

The present invention will be described in detail below.

The invention provides a flavonoid compound with a structure shown as the following formula (I) and pharmaceutically acceptable salt thereof or solvate or hydrate thereof:

wherein,

x and Y are each independently selected from O, S or NH;

R₁is C with or without substituents₃-C₁₅A linear or branched saturated or unsaturated hydrocarbon group;

R₂and R₄Each independently of the otherIs selected from the group consisting of hydrogen, trifluoromethyl, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated hydrocarbon group, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated alkanoyl, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated alkylsulfonyl, substituted or unsubstituted aromatic cyclylsulfonyl or aromatic heterocyclylsulfonyl, substituted or unsubstituted aromatic ring group or aromatic heterocyclic group, substituted or unsubstituted aromatic (C1-C4 linear or branched saturated or unsaturated) alkanoyl;

R₃and R₅Each independently selected from the group consisting of hydrogen, halogen, cyano, trifluoromethyl, hydroxy, mercapto, amino, substituted or unsubstituted C1-C6 straight or branched chain saturated or unsaturated alkoxy, substituted or unsubstituted C1-C6 straight or branched chain saturated or unsaturated alkylthio, substituted or unsubstituted C1-C6 straight or branched chain saturated or unsaturated alkylamino, substituted or unsubstituted C1-C6 straight or branched chain saturated or unsaturated hydrocarbyl;

R₄and R₅In addition to being each independently selected from the above groups, may also be each independently

Or- (CH)₂CH ═ CH) -, to form a cyclic structure;

R₆～R₁₀each independently selected from hydrogen, hydroxyl, halogen atom, sulfhydryl, nitro, cyano, amino, trifluoromethyl, hydroxymethyl, carboxyl, C1-C6 linear or branched saturated or unsaturated alkyl, C1-C6 linear or branched saturated or unsaturated alkoxy, C1-C6 linear or branched saturated or unsaturated alkylamino;

the aromatic ring group in the aromatic ring group sulfonyl and aromatic ring group is phenyl, tolyl, xylyl, biphenyl, naphthyl, indenyl, anthryl or phenanthryl;

the aromatic heterocyclic radical sulfonyl and the aromatic heterocyclic radical in the aromatic heterocyclic radical are furyl, thienyl, pyrrolyl, imidazolyl, thiazolyl, pyrazolyl, isothiazolyl, isoxazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazolyl or tetrazolyl;

the aryl (C1-C4 linear or branched chain saturated or unsaturated) alkanoyl is the combination of the aromatic cyclic group described in the specification and C1-C4 linear or branched chain saturated or unsaturated alkanoyl, wherein the preferable combination is benzoyl, phenylacetyl, phenylpropanoyl, phenylbutyryl, (Z) styroyl, p-phenylbenzoyl, p-phenylphenylacetyl, p-phenylphenylpropionyl and p-phenylphenylbutyryl;

the substituent is one or more selected from the group consisting of a halogen atom, a cyano group, a nitro group, an amino group, a hydroxyl group, a hydroxymethyl group, a trifluoromethyl group, a trifluoromethoxy group, a carboxyl group, a mercapto group, a C1-C4 linear or branched saturated or unsaturated alkyl group, a C1-C4 linear or branched saturated or unsaturated alkoxy group, a C1-C4 linear or branched saturated or unsaturated alkanoyl group, and a C1-C4 linear or branched saturated or unsaturated alkylamino group.

The compound represented by the formula (I) of the present invention is preferably a flavonoid compound as follows and a pharmaceutically acceptable salt thereof or a solvate or hydrate thereof:

wherein when X is O and Y is O,

R₁is C with or without substituents₃-C₁₅Linear or branched, saturated or unsaturated hydrocarbon radicals, the preferred radicals being 3-methylbut-2-enyl or 5-methylhexa-2, 4-dienyl;

R₂and R₄Each is independentSelected from the group consisting of hydrogen, trifluoromethyl, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated hydrocarbon radical, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated alkanoyl, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated alkylsulfonyl, substituted or unsubstituted aromatic cyclylsulfonyl or aromatic heterocyclylsulfonyl, substituted or unsubstituted aromatic ring radical or aromatic heterocyclic radical, substituted or unsubstituted aromatic (C1-C4 linear or branched saturated or unsaturated) alkanoyl;

Or- (CH)₂CH ═ CH) -, to form a cyclic structure;

the aromatic ring group in the aromatic ring sulfonyl and aromatic ring group, the aromatic heterocyclic group in the aromatic heterocyclic group, the aromatic (C1-C4 straight chain or branched chain saturated or unsaturated) alkanoyl and the substituent are as described in the specification.

Representative examples of the compounds represented by the formula (I) of the present invention are as follows:

1)5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2-phenyl-4H-chromen-4-one;

2)5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

3) 5-hydroxy-3- (3-methylbut-2-enyl) -4-oxo-2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-7-substituted methanesulfonate;

4) 5-hydroxy-7- (2-methylbenzyloxy) -3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

5)7- (cyclohexylmethoxy) -5-hydroxy-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

6) 5-hydroxy-3- (3-methylbut-2-enyl) -7-propargyloxy-2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

7) 5-hydroxy-3- (3-methylbut-2-enyl) -7- (2-morpholin-N-substituted ethoxy) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

8) 5-hydroxy-3- (3-methylbut-2-enyl) -7- (2-N-tert-butoxycarbonylpiperazine-N' -substituted ethoxy) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

9) 5-hydroxy-8, 8-dimethyl-3- (3-methylbut-2-enyl) -2-phenyl-8H-pyran [2, 3-f ] chromen-4-one;

10) 5-hydroxy-8, 8-dimethyl-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [2, 3-f ] chromen-4-one;

11)8, 8-dimethyl-3- (3-methylbut-2-enyl) -4-oxo-2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [2, 3-f ] chromen-5-substituted acetate;

12) 5-hydroxy-8, 8-dimethyl-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [3, 2-g ] chromen-4-one;

13) (E) -5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-chromen-4-one;

14) (E) -5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one;

15) (E) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -4-oxo-2-phenyl-4H-benzopyran-7-substituted methanesulfonate;

16) (E) -7- (cyclohexylmethoxy) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-chromen-4-one;

17) (E) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -7- (2-morpholine-N-substituted ethoxy) -2-phenyl-4H-chromen-4-one;

18) (E) -5-hydroxy-7- (2-methylbenzyloxy) -3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-chromen-4-one;

19) (E) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-7-propargyloxy-4H-chromen-4-one;

20) (E) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -4-oxo-2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-7-substituted methanesulfonate;

21) (E) -7- (cyclohexylmethoxy) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

22) (E) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -7- (2-morpholine-N-substituted ethoxy) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

23) (E) -5-hydroxy-7- (2-methylbenzyloxy) -3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

24) (E) -5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -7-propargyloxy-2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

25) (E) -5-hydroxy-8, 8-dimethyl-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-8H-pyran [2, 3-f ] chromen-4-one;

26) (E) -5-hydroxy-8, 8-dimethyl-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [2, 3-f ] chromen-4-one;

27) (E) -7-cyclopropylmethoxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-chromen-4-one;

28) (E) -7-cyclopropylmethoxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

29) (E) -7-allyloxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-chromen-4-one;

30) (E) -7-allyloxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one;

31)5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (3, 4-dimethoxyphenyl) -4H-chromen-4-one;

32) (E) -5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (3, 4-dimethoxyphenyl) -4H-chromen-4-one;

33)5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (3, 4-dichlorophenyl) -4H-chromen-4-one;

34) (E) -5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (3, 4-dichlorophenyl) -4H-chromen-4-one;

35)5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (3-dimethylaminophenyl) -4H-chromen-4-one;

36) (E) -5, 7-dihydroxy-3- (5-methylhexane-2, 4-dienyl) -2- (3-dimethylaminophenyl) -4H-benzopyran-4-one.

The method for preparing the flavonoid compound shown in the formula (I) can be respectively carried out according to the following two processes:

scheme I:

the following examples are given: a.R₁X (X ═ Cl or Br), a base, an organic solvent; b. phloroglucinol, microwave; 3-methyl-2-butenal, an organic solvent, and microwave or heating; d.R₄X (X ═ Cl or Br) or R₄OSO₂CH₃Base, organic solvent; e.R₂X (X ═ Cl or Br) or R₂OSO₂CH₃Base, organic solvent.

The preparation process of scheme I is specifically illustrated below:

(1) r of the formula shown in scheme I₁、R₂、R₄、R₆-R₁₀Selected from the ranges as described before in the specification;

(2) reference [ Seijas, j.a.; v zqueq-Tato, M.P.; carbollido-Reboredo, r.j.org.chem.2005, 70, 2855]As shown in step b, R₁The substituted 3-oxo ethyl phenylpropionate and phloroglucinol react for 3 to 60 minutes at 50 to 150 ℃ under the action of microwaves to prepare a 3-substituted 5, 7-dihydroxy flavone derivative;

(3) the step c can be prepared by adopting two methods of microwave or heating reaction, wherein the microwave reaction adopts a closed system, the reaction temperature can be from 80 ℃ to 200 ℃, and the reaction time is from 15 minutes to 20 hours; the heating reaction adopts a drying system, the reaction temperature can be from 80 ℃ to 200 ℃, and the reaction time is from 10 hours to 7 days;

(4) the base used in steps a, d and e is selected from organic bases including pyridine, triethylamine, 4-Dimethylaminopyridine (DMAP), diisopropylethylamine, potassium tert-butoxide and inorganic bases including sodium carbonate, potassium carbonate, cesium carboxylate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride;

(5) the organic solvent used in steps a, c, d and e can be anhydrous inert solvent including diethyl ether, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, N-methylpyrrolidone, pyridine, dichloromethane, chloroform, N-hexane, benzene and toluene; wherein the most suitable solvent for steps a and d is anhydrous tetrahydrofuran or anhydrous DMF; the most suitable solvent for step c is anhydrous pyridine.

Scheme II:

the following examples are given: momcl, acetone; b.R₆-R₁₀Substituted benzoyl chloride and pyridine; c. base, organic solvent; d.R₆-R₁₀Substituted benzoyl chloride, alkali and an organic solvent; e.R₁X (X ═ Cl or Br), a base, an organic solvent; f. acid, organic solvent; 3-methyl-2-butenal, an organic solvent, and microwave or heating; h.R₄X (X ═ Cl or Br) or R₄OSO₂CH₃Base, organic solvent; i.R₂X (X ═ Cl or Br) or R₂OSO₂CH₃Base, organic solvent.

The preparation process of scheme II is specifically illustrated below:

(1) r in the formula shown in scheme II₁-R₁₀Selected from the ranges as described before in the specification;

(2) preparing a desired diketone intermediate by base rearrangement as shown in steps c and d, wherein the base used in step c is selected from the group consisting of organic bases including 4-Dimethylaminopyridine (DMAP), sodium methoxide, sodium ethoxide, potassium tert-butoxide, and inorganic bases including sodium hydroxide, potassium hydroxide, sodium hydride; the base used in step d is selected from organic bases including sodium methoxide, sodium ethoxide and potassium tert-butoxide and inorganic bases including sodium hydroxide, potassium hydroxide and sodium hydride;

(3) step g, microwave or heating reaction can be adopted, wherein a closed system is adopted for microwave reaction, the reaction temperature can be from 80 ℃ to 200 ℃, and the reaction time is from 15 minutes to 20 hours; a drying system is adopted for heating reaction, the reaction temperature is 80-200 ℃, and the reaction time is 10 hours-7 days;

(4) step f, preparing a substituted 5, 7-dihydroxyflavone intermediate by using acid deprotection and cyclization, wherein the reaction temperature is controlled to be 0-100 ℃, and the acid is 0.5-6M hydrochloric acid, sulfuric acid or glacial acetic acid;

(5) the base used in steps e, h and i is selected from organic bases including pyridine, triethylamine, 4-Dimethylaminopyridine (DMAP), diisopropylethylamine, potassium tert-butoxide and inorganic bases including sodium carbonate, potassium carbonate, cesium carboxylate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride;

(6) the organic solvent used in step c-i may be an anhydrous inert solvent such as diethyl ether, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, N-methylpyrrolidone, pyridine, dichloromethane, chloroform, N-hexane, benzene, toluene, etc., depending on the nature of the specific compound; wherein the most suitable solvent for steps e and h is anhydrous tetrahydrofuran or anhydrous DMF; the most suitable solvent for step g is anhydrous pyridine.

The method for purifying the crude product obtained in each step in the reaction flow I and the reaction flow II comprises column chromatography or recrystallization, wherein the recrystallization can be performed by using methanol, ethanol, isopropanol, acetone, ethyl acetate, chloroform, dichloromethane, toluene and n-hexane, or by using a mixed solvent consisting of two or more components according to a proper proportion.

The compounds prepared according to reaction scheme I and scheme II can be subjected to salt-forming reaction as required.

Another aspect of the present invention relates to a pharmaceutical composition for preventing and/or treating inflammatory diseases, immune diseases such as asthma and allergy, and cardiovascular diseases, wherein the pharmaceutical composition comprises a substituted flavonoid compound represented by formula (I) or a pharmaceutically acceptable salt thereof, or a solvate or hydrate thereof, and at least one pharmaceutically acceptable carrier, and the pharmaceutical composition can be used for in vivo treatment and has biocompatibility. The pharmaceutical composition may be prepared in various forms according to different administration routes. The pharmaceutical composition can be used for preventing and/or treating inflammatory diseases, immunological diseases such as asthma and allergy, and cardiovascular diseases.

The pharmaceutical composition for preventing and/or treating inflammatory diseases, immunological diseases such as asthma and allergy and cardiovascular diseases, which is disclosed by the invention, is a pharmaceutical composition comprising an effective dose of a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a hydrate thereof, and one or more suitable pharmaceutically acceptable carriers. Pharmaceutically acceptable carriers herein include, but are not limited to: ion exchangers, alumina, aluminium stearate, lecithin, serum proteins, phosphate buffer substances, glycerol, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, beeswax and lanolin.

Pharmaceutical compositions of the compounds of the present invention may be administered in any of the following ways: oral, aerosol inhalation, rectal, nasal, buccal, topical, parenteral, e.g. subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or via an external reservoir. Wherein oral or intramuscular injection, intraperitoneal or intravenous administration is preferred for treating inflammation.

It is further noted that the dosage and method of administration of the pharmaceutical compositions of the compounds of the present invention will depend upon a variety of factors including the age, body weight, sex, physical condition, nutritional status, the strength of the activity of the compound, the time of administration, the rate of metabolism, the severity of the condition, and the subjective judgment of the treating physician. The recommended dosage is, for example, 5mg to 10mg/kg daily at the beginning, and the maintenance dose can be reduced to 3mg/kg daily. And (3) capsule preparation: 0.25 g/pellet; injection liquid: 0.25g/5 ml; oral solution: 5g/50 ml.

Detailed Description

The preparation of the compounds of the formula (I) is illustrated below by way of examples, which are not to be construed as limiting the invention in any way. All parameters and associated descriptions in the examples are by mass unless otherwise indicated.

In the following preparation examples, the melting point was measured by an X-4 type melting point apparatus manufactured by Shanghai precision scientific instruments Co., Ltd without correcting the temperature; the nuclear magnetic resonance is measured by a Bruker AMX-300/400 type nuclear magnetic resonance instrument and an INVOA-600 type nuclear magnetic resonance instrument, TMS is an internal standard, and the chemical shift unit is ppm; mass spectra were determined by MAT-711 and MAT-95 mass spectrometers; silica gel 200-mesh and 300-mesh for column chromatography, produced in Qingdao ocean factory; the TLC silica gel plate is an HSGF-254 thin-layer chromatography prefabricated plate produced by a cigarette bench chemical plant; the boiling range of petroleum ether is 60-90 ℃; and developing with ultraviolet lamp and iodine cylinder. In the following statement, "concentration" (if the operation method is not particularly specified) means distilling off the solvent in the solution of the preparation compound by means of a rotary evaporator; "drying" (if the procedure is not specified) means drying the preparation compound at 30-80 ℃ in a DHG-9240A oven.

The structures of the compounds described in the examples of the present invention are shown in Table 1.

Structures of the compounds in the examples described in Table 1

Example 1:

synthesis of 5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2-phenyl-4H-chromen-4-one (Compound 1):

step (1): 0.5g of ethyl 3-oxo-3-phenylpropionate was dissolved in 5mL of DMF to give a colorless clear solution, and 0.4g of potassium carbonate and 0.305mL of 3, 3-dimethylallyl bromide were added thereto with stirring at room temperature to react overnight at room temperature. The next day, 20mL of water was added to the system, transferred to a separatory funnel, extracted three times with 30mL of ethyl acetate, and combinedThe organic phase was washed twice with 25mL of water and twice with 20mL of saturated brine, and dried over anhydrous magnesium sulfate. Filtering to remove desiccant, evaporating to remove solvent to obtain crude viscous oily substance, and purifying by column chromatography to obtain pure product 575mg with yield of 85%.¹H NMR(CDCl₃，300M)：δ8.24(d，J＝7.2Hz，2H)，7.57(t，J＝7.2Hz，1H)，7.45(t，J＝7.5Hz，2H)，5.10(t，J＝7.2Hz，1H)，4.29(t，J＝7.5Hz，1H)，4.13(q，J＝7.2Hz，2H)，2.7(m，1H)，1.64(s，3H)，1.62(s，3H)，1.16(t，J＝7.2Hz，3H)。

Step (2): 0.5g of the product is evenly mixed with 243mg of phloroglucinol and then placed in a household microwave oven under 320W for reaction for 5-20 minutes. The reaction system is cooled to room temperature and then dissolved in 10mL of ethyl acetate, insoluble substances are removed by filtration, and a pure product of 167mg is obtained by mother liquor column chromatography purification with the yield of 27%.¹H NMR(CDCl₃，300M)：δ13.02(s，1H)，7.46-7.61(m，5H)，6.35(s，1H)，6.30(s，1H)，5.20(t，J＝6.0Hz，1H)，3.19(d，J＝6.0Hz，2H)，1.69(s，3H)，1.53(s，3H)。LRMS(EI)m/z 322(M⁺)，279(100％)。

Example 2:

synthesis of 5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one (Compound 2):

step (1): bis-MOM protected trihydroxyacetophenone was prepared according to literature procedure [ Kumazawa, t.et al. carbohydr.res.2000, 329, 507 ].

Step (2): 3.1g of bis-MOM protected trihydroxyacetophenone was dissolved in 20mL of anhydrous DMF, and 1.5g of sodium hydride solution in DMF was slowly dropped under cooling in ice bath, and 3.0g of 2, 4, 5-trimethoxybenzoyl chloride was added thereto, followed by reaction at room temperature overnight. The system is added with 50mL of cold water with the temperature of 0-10 ℃, 50mL of ethyl acetate is used for extraction, organic phases are combined, washed by 50mL of water and 50mL of saturated saline solution, and dried by anhydrous magnesium sulfate. The drying agent is filtered off, and the product obtained by evaporating the solvent can be directly used for the next reaction, with the yield of 97%.¹H NMR(CDCl₃，300M)：δ13.42(s，1H)，7.56(s，1H)，6.48(s，1H)，6.29(s，1H)，6.25(s，1H)，5.17(s，2H)，5.05(s，2H)，4.59(s，3H)，3.96(s，3H)，3.88(s，3H)，3.77(s，3H)，3.47(s，3H)，3.34(s，3H)。

And (3): dissolving 4g of the product of the step (2) in 40ml of DMF, adding 1.4g of anhydrous potassium carbonate, injecting 1.2ml of 3, 3-dimethylallyl bromide under stirring, reacting overnight at room temperature until the raw material disappears, adding 100ml of water, extracting twice with 80ml of ethyl acetate, combining organic phases, washing twice with 80ml of water, washing twice with 80ml of saturated saline solution, and drying with anhydrous magnesium sulfate. Filtering to remove desiccant, evaporating to remove solvent to obtain crude viscous oily substance, and purifying by column chromatography to obtain pure product 3.5g with yield of 76%.¹H NMR(CDCl₃，300M)：δ13.67(s，1H)，7.60(s，1H)，6.47(s，1H)，6.29(s，1H)，5.49(t，J＝6.3Hz，1H)，5.16(s，2H)，5.08(t，J＝5.7Hz，1H)，4.97(q，J＝6.9Hz，1H)，3.95(s，3H)，3.87(s，3H)，3.73(s，3H)，3.46(s，3H)，3.26(s，3H)，2.64(q，J＝5.7Hz，2H)，1.66(s，3H)，1.62(s，3H)。

And (4): and (3) dissolving 1.2g of the product obtained in the step (3) in 50mL of methanol, dropwise adding 1mL of concentrated sulfuric acid, heating at 50-100 ℃ for reacting for 15-60 minutes, cooling, separating out light yellow needle crystals from a reaction system, filtering, and washing with 10mL of methanol to obtain 1.1g of a pure product with the yield of 77%.¹H NMR(CDCl₃，300M)：δ13.07(s，1H)，6.84(s，1H)，6.60(s，1H)，6.29(s，1H)，6.26(s，1H)，5.11(t，J＝6.3Hz，1H)，3.96(s，3H)，3.83(s，3H)，3.80(s，3H)，3.03(d，J＝6.3Hz，2H)，1.61(s，3H)，1.41(s，3H).LRMS(EI)m/z412(M⁺)，369(100％)。

Example 3:

synthesis of 5-hydroxy-3- (3-methylbut-2-enyl) -4-oxo-2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-7-substituted methanesulfonate (Compound 3):

dissolving 100mg of compound 2 in 2mL of anhydrous tetrahydrofuran, adding 20 mu L of methanesulfonyl chloride and 34 mu L of triethylamine under ice-bath cooling, reacting at room temperature until the raw materials disappear, evaporating to remove the solvent, adding 10mL of water into the system, extracting with 15mL of ethyl acetate, and sequentially adding 20mL of water and saturated salt into the organic phaseThe reaction mixture was washed twice with 20mL of water and dried over anhydrous magnesium sulfate. Filtering to remove desiccant, evaporating to remove solvent to obtain crude product, and recrystallizing with mixed solvent of ethyl acetate and petroleum ether (V: 1/3) to obtain pure product 107mg with yield of 90%.¹H NMR(CDCl₃，300M)：δ13.15(s，1H)，6.86(s，1H)，6.84(s，1H)，6.66(s，1H)，6.60(s，1H)，5.07(t，J＝6.6Hz，1H)，3.97(s，3H)，3.85(s，3H)，3.79(s，3H)，3.20(s，3H)，3.05(d，J＝6.6Hz，2H)，1.61(s，3H)，1.42(s，3H).LRMS(EI)m/z 490(M⁺)，447(100％)。

Example 4:

synthesis of 5-hydroxy-7- (2-methylbenzyloxy) -3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (Compound 4):

the title compound was obtained in 61% yield as a white solid by the same operations and workup as in example 3, except that 2-methylbenzyl chloride was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran.¹H NMR(CDCl₃，300M)：δ13.00(s，1H)，7.21-7.38(m，4H)，6.84(s，1H)，6.60(s，1H)，6.43(s，2H)，5.11(t，J＝6.6Hz，1H)，5.06(s，2H)，3.96(s，3H)，3.84(s，3H)，3.80(s，3H)，3.03(d，J＝6.6Hz，2H)，2.35(s，3H)，1.62(s，3H)，1.41(s，3H)。LRMS(EI)m/z 516(M⁺)，485(100％)。

Example 5:

synthesis of 7- (cyclohexylmethoxy) -5-hydroxy-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one (Compound 5):

the title compound was obtained in 75% yield as a white solid by the same operations and workup as in example 3, except that cyclohexylmethyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran.¹H NMR(CDCl₃，300M)：δ12.96(s，1H)，6.84(s，1H)，6.60(s，1H)，6.32(s，2H)，5.11(t，J＝6.3Hz，1H)，3.97(s，3H)，3.84(s，3H)，3.79(s，3H)，3.77(d，J＝6.0Hz，2H)，3.03(d，J＝6.3Hz，2H)，1.63-1.85(m，6H)，1.61(s，3H)，1.41(s，3H)，1.00-1.38(m，5H).LRMS(EI)m/z 508(M⁺)，465(100％)。

Example 6:

synthesis of 5-hydroxy-3- (3-methylbut-2-enyl) -7-propargyloxy-2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (Compound 6):

the title compound was obtained in 72% yield as a white solid by the same operations and workup as in example 3, except that propargyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran.¹H NMR(CDCl₃，300M)：δ6.84(s，1H)，6.60(s，1H)，6.42(d，J＝2.1Hz，1H)，6.40(d，J＝2.1Hz，1H)，5.10(t，J＝6.3Hz，1H)，4.71(d，J＝2.1Hz，1H)，3.96(s，3H)，3.83(s，3H)，3.79(s，3H)，3.03(d，J＝6.3Hz，2H)，2.54(t，J＝2.1Hz，1H)，1.61(s，3H)，1.41(s，3H)。LRMS(EI)m/z450(M⁺)，407(100％)。

Example 7:

synthesis of 5-hydroxy-3- (3-methylbut-2-enyl) -7- (2-morpholin-N-substituted ethoxy) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one (Compound 7):

the title compound was obtained in 65% yield by the same operations and workup as in example 3 except that 2-morpholine-N-substituted ethylmethanesulfonate was used instead of methanesulfonyl chloride, cesium carboxylate was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran.¹H NMR(CDCl₃，300M)：δ12.98(s，1H)，6.83(s，1H)，6.59(s，1H)，6.33(s，2H)，5.10(t，J＝6.3Hz，1H)，4.13(t，J＝5.4Hz，2H)，3.96(s，3H)，3.84(s，3H)，3.79(s，3H)，3.72(t，J＝4.5Hz，4H)，3.05(d，J＝6.3Hz，2H)，2.80(t，J＝5.4Hz，1H)，2.56(t，J＝4.5Hz，4H)，1.61(s，3H)，1.41(s，3H)。LRMS(EI)m/z525(M⁺)，100(100％)。

Example 8

Synthesis of 5-hydroxy-3- (3-methylbut-2-enyl) -7- (2-N-tert-butoxycarbonylpiperazine-N' -substituted ethoxy) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one (Compound 8):

the title compound was obtained in 63% yield as a white solid by the same operations and workup as in example 3, except that tert-butyl 4- (2-methanesulfonyloxyethyl) piperazine-1-carboxylate was used instead of methanesulfonyl chloride, cesium carboxylate was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran.¹H NMR(CDCl₃，300M)：δ12.99(s，1H)，6.84(s，1H)，6.60(s，1H)，6.34(s，2H)，5.10(t，J＝6.6Hz，1H)，4.12(t，J＝6.0Hz，2H)，3.96(s，3H)，3.83(s，3H)，3.80(s，3H)，3.44(t，J＝4.5Hz，4H)，3.03(d，J＝6.6Hz，2H)，2.81(t，J＝5.7Hz，1H)，2.50(t，J＝4.5Hz，4H)，1.61(s，3H)，1.45(s，9H)，1.41(s，3H)。LRMS(EI)m/z 624(M⁺)，143(100％)。

Example 9

Synthesis of 5-hydroxy-8, 8-dimethyl-3- (3-methylbut-2-enyl) -2-phenyl-8H-pyran [2, 3-f ] chromen-4-one (Compound 9):

0.1g of the compound 1 is dissolved in 1ml of pyridine, 35 mu L of 3-methyl-2-butenal is added, and the closed system is placed in a microwave reactor and reacts for 5 to 15 hours at the temperature of 120-. The reaction system is cooled to room temperature, the solvent is evaporated, and the residue is purified by column chromatography to obtain a pure solid product 64mg with a yield of 53%.¹H NMR(CDCl₃，300M)：δ13.02(s，1H)，7.48-7.64(m，5H)，6.63(d，J＝10.2Hz，1H)，6.27(s，1H)，5.51(d，J＝10.2Hz，1H)，5.21(t，J＝6.3Hz，1H)，3.21(d，J＝6.3Hz，2H)，1.70(s，3H)，1.57(s，3H)，1.45(s，6H).LRMS(EI)m/z 388(M⁺)，373(100％)。

Example 10

Synthesis of 5-hydroxy-8, 8-dimethyl-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [2, 3-f ] chromen-4-one (Compound 10):

the title compound was obtained as a pale yellow solid in 51% yield by the same procedures and post-treatment as in example 9, except that the compound 2 was used in place of the compound 1 (used amount was not changed).¹H NMR(CDCl₃，300M)：δ13.10(s，1H)，6.87(s，1H)，6.60(s，1H)，6.57(d，J＝9.9Hz，1H)，6.25(s，1H)，5.47(d，J＝9.9Hz，1H)，5.11(t，J＝6.6Hz，1H)，3.97(s，3H)，3.84(s，3H)，3.81(s，3H)，3.04(d，J＝6.6Hz，2H)，1.61(s，3H)，1.44(s，6H)，1.41(s，3H).LRMS(EI)m/z 478(M⁺)，463(100％)。

Example 11

Synthesis of 8, 8-dimethyl-3- (3-methylbut-2-enyl) -4-oxo-2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [2, 3-f ] chromene-5-substituted acetate (Compound 11):

dissolving 100mg of compound 10 in 2ml of pyridine, dripping 100 mu L of acetic anhydride, reacting at room temperature for 0.5-3 hours, evaporating to remove the solvent, and purifying the residue by column chromatography to obtain a pure solid product 94mg with the yield of 86%.¹H NMR(CDCl₃，300M)：δ6.84(s，1H)，6.65(d，J＝9.9Hz，1H)，6.59(s，1H)，6.45(s，1H)，5.57(d，J＝9.9Hz，1H)，5.10(t，J＝6.0Hz，1H)，3.96(s，3H)，3.81(s，3H)，3.78(s，3H)，2.99(d，J＝6.0Hz，2H)，2.45(s，3H)，1.57(s，3H)，1.46(s，6H)，1.32(s，3H)。LRMS(EI)m/z520(M⁺)，463(100％)。

Example 12

Synthesis of 5-hydroxy-8, 8-dimethyl-3- (3-methylbut-2-enyl) -2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [3, 2-g ] chromen-4-one (Compound 12):

0.2g of the compound 2 was dissolved in 2ml of pyridine, 68. mu.L of 3-methyl-2-butenal was added, and the reaction was carried out at 200 ℃ under nitrogen for 3 to 7 days until the reaction did not proceed any more. Cooling the reaction system to room temperature, evaporating the solvent,purifying the residue by column chromatography to obtain a pure solid product 25mg with a yield of 11%.¹H NMR(CDCl₃，300M)：δ13.31(s，1H)，6.83(s，1H)，6.73(d，J＝10.2Hz，1H)，6.59(s，1H)，6.26(s，1H)，5.54(d，J＝9.9Hz，1H)，5.11(t，J＝6.0Hz，1H)，3.96(s，3H)，3.83(s，3H)，3.79(s，3H)，3.02(d，J＝6.0Hz，2H)，1.61(s，3H)，1.44(s，6H)，1.40(s，3H)。LRMS(EI)m/z 478(M⁺)，463(100％)。

Example 13

(E) Synthesis of (E) -5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-benzopyran-4-one (Compound 13).

The title compound was obtained by the same procedures and post-treatment as in example 1 except for using (E) -1-bromo-5-methylhexane-2, 4-diene in place of 3, 3-dimethylallyl bromide (used in a constant amount).

Example 14

(E) -synthesis of 5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (compound 14):

the title compound was obtained by the same operations and post-treatment as in example 2, except that (E) -1-bromo-5-methylhexane-2, 4-diene was used instead of 3, 3-dimethylallyl bromide (used in a constant amount).

Example 15

(E) -Synthesis of 5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -4-oxo-2-phenyl-4H-benzopyran-7-substituted methanesulfonate (Compound 15):

the title compound was obtained by following the same procedures and post-treatment as in example 3, except that compound 13 was used instead of compound 2 (used in the same amount).

Example 16

(E) -Synthesis of 7- (cyclohexylmethoxy) -5-hydroxy-3- (5-methylhexane-2, 4-dienyl) -2-phenyl-4H-benzopyran-4-one (Compound 16):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 13 was used instead of compound 2, cyclohexylmethyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 17

(E) -Synthesis of 5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -7- (2-morpholin-N-substituted ethoxy) -2-phenyl-4H-benzopyran-4-one (Compound 17):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 13 was used instead of compound 2, 2-morpholine-N-substituted ethylmethanesulfonate was used instead of methanesulfonyl chloride, cesium carboxylate was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in the same amount).

Example 18

(E) -Synthesis of 5-hydroxy-7- (2-methylbenzyloxy) -3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-benzopyran-4-one (Compound 18):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 13 was used instead of compound 2, 2-methylbenzyl chloride was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 19

(E) -Synthesis of 5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-7-propargyloxy-4H-benzopyran-4-one (Compound 19):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 13 was used instead of compound 2, propargyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 20

(E) -Synthesis of 5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -4-oxo-2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-7-substituted methanesulfonate (Compound 20):

the title compound was obtained by following the same procedures and post-treatment as in example 3, except that compound 14 was used instead of compound 2 (used in the same amount).

Example 21

(E) -Synthesis of 7- (cyclohexylmethoxy) -5-hydroxy-3- (5-methylhexane-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (Compound 21):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 14 was used instead of compound 2, cyclohexylmethyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 22

(E) -Synthesis of 5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -7- (2-morpholine-N-substituted ethoxy) -2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (Compound 22):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 14 was used instead of compound 2, 2-morpholine-N-substituted ethylmethanesulfonate was used instead of methanesulfonyl chloride, cesium carboxylate was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in the same amount).

Example 23

(E) -Synthesis of 5-hydroxy-7- (2-methylbenzyloxy) -3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (Compound 23):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 14 was used instead of compound 2, 2-methylbenzyl chloride was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 24

(E) -Synthesis of 5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -7-propargyloxy-2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (Compound 24):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 14 was used instead of compound 2, propargyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 25

(E) -Synthesis of 5-hydroxy-8, 8-dimethyl-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-8H-pyran [2, 3-f ] chromen-4-one (Compound 25):

the title compound was obtained by the same procedures and post-treatment as in example 9, except that compound 13 was used instead of compound 1 (used amount was not changed).

Example 26

(E) -Synthesis of 5-hydroxy-8, 8-dimethyl-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -8H-pyran [2, 3-f ] chromen-4-one (Compound 26):

the title compound was obtained by following the same procedures and post-treatment as in example 9, except that compound 14 was used instead of compound 1 (used amount was not changed).

Example 27

(E) -Synthesis of 7-cyclopropylmethoxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-chromen-4-one (Compound 27):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 13 was used instead of compound 2, cyclopropylmethyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 28

(E) -Synthesis of 7-cyclopropylmethoxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-chromen-4-one (Compound 28):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 14 was used instead of compound 2, cyclopropylmethyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in an unchanged amount).

Example 29

(E) -Synthesis of 7-allyloxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2-phenyl-4H-benzopyran-4-one (Compound 29):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 13 was used instead of compound 2, allyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in a constant amount).

Example 30

(E) -synthesis of 7-allyloxy-5-hydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (2, 4, 5-trimethoxyphenyl) -4H-benzopyran-4-one (compound 30):

the title compound was obtained by following the same procedures and workup as in example 3 except that compound 14 was used instead of compound 2, allyl bromide was used instead of methanesulfonyl chloride, sodium hydride was used instead of triethylamine, and anhydrous DMF was used instead of anhydrous tetrahydrofuran (used in a constant amount).

Example 31

Synthesis of 5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (3, 4-dimethoxyphenyl) -4H-chromen-4-one (Compound 31):

the title compound was obtained by following the same procedures and post-treatment as in example 2, except that 3, 4-trimethoxybenzoyl chloride was used instead of 2, 4, 5-trimethoxybenzoyl chloride (used in the same amount).

Example 32

(E) -Synthesis of 5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (3, 4-dimethoxyphenyl) -4H-benzopyran-4-one (Compound 32):

the title compound was obtained by following the same procedures and post-treatment as in example 2 except for using 3, 4-trimethoxybenzoyl chloride instead of 2, 4, 5-trimethoxybenzoyl chloride and (E) -1-bromo-5-methylhexa-2, 4-diene instead of 3, 3-dimethylallyl bromide (used in the same amount).

Example 33

Synthesis of 5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (3, 4-dichlorophenyl) -4H-benzopyran 4-one (Compound 33):

the title compound was obtained by the same operations and post-treatment as in example 2, except for using 3, 4-dichlorobenzoyl chloride instead of 2, 4, 5-trimethoxybenzoyl chloride (used in the same amount).

Example 34

(E) -synthesis of 5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (3, 4-dichlorophenyl) -4H-benzopyran-4-one (compound 34):

the title compound was obtained by following the same procedures and post-treatment as in example 2 except for using 3, 4-dichlorobenzoyl chloride instead of 2, 4, 5-trimethoxybenzoyl chloride and (E) -1-bromo-5-methylhexa-2, 4-diene instead of 3, 3-dimethylallyl bromide (used in the same amount).

Example 35

Synthesis of 5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2- (3-dimethylaminophenyl) -4H-benzopyran-4-one (Compound 35):

the title compound was obtained by the same operations and post-treatment as in example 2, except for using 3-dimethylaminobenzoyl chloride instead of 2, 4, 5-trimethoxybenzoyl chloride (used in the same amount).

Example 36

(E) -synthesis of 5, 7-dihydroxy-3- (5-methylhexa-2, 4-dienyl) -2- (3-dimethylaminophenyl) -4H-benzopyran-4-one (compound 36):

the title compound was obtained by following the same operations and post-treatment as in example 2, except for using 3-dimethylaminobenzoyl chloride instead of 2, 4, 5-trimethoxybenzoyl chloride and (E) -1-bromo-5-methylhexa-2, 4-diene instead of 3, 3-dimethylallyl bromide (used in the same amount).

Test examples:

isolated rat neutrophils Release leukotriene B₄Inhibition test of

Leukotriene B₄Is one of the products of the metabolic pathway of arachidonic acid 5-lipoxygenase, and is involved in many physiological and pathological processes of the body, including cell chemotaxis and the like under inflammatory diseases, and the neutrophil is LTB₄The main source of synthesis and release. The study was conducted to see if the series of compounds released LTB on isolated rat neutrophils₄Has inhibiting effect and corresponding inhibiting strength.

1 materials of the experiment

1.1 test drugs: all tested compounds are provided by Shanghai pharmaceutical research institute of Chinese academy of sciences. All compounds were dissolved in DMSO at 0.05M stock concentration and diluted to the corresponding test concentrations with Hanks balanced salt buffer just prior to use.

1.2 Experimental reagents: glycogen type II (Sigma-Aldrich Co, 10K154), indomethacin (Sigma-Aldrich Co, 061K1368), A23187(Sigma-Aldrich Co), L-cysteine (Shanghai Kangda amino acid works, lot number: 20030601), LTB₄EIA assay kit (Cayman Chemical Company, lots: 135349, 137768, 124376, respectively).

1.3 test animals: SD rats, clean grade, male and female irresistible, body weight 200 + -20 g, provided by the Experimental animals center of the medical college of Zhejiang university.

1.4 Experimental instruments: microplate reader (Thermo, Multiskan spec), thermostated water bath, microscope, etc.

2 method of experiment

2.1 preparation of rat leukocyte suspension

Taking normal rats, carrying out intraperitoneal injection (ip) on 20mL/kg of 0.2% glycogen, killing the rats by femoral artery exsanguination after 16h, carrying out intraperitoneal lavage on each rat by 10mL of Hanks balanced salt buffer solution, collecting the peritoneal lavage solution, centrifuging for 10min at 4 ℃, 2000r/min, adding 5mL of 0 ℃ refrigerated distilled water into precipitated cells to dissolve red blood cells, immediately adding 1.8% sodium chloride solution with the same volume after 1min, centrifuging for 5min at 2000r/min (4 ℃), suspending and washing the precipitated cells for 2 times by 5mL of Hanks balanced salt buffer solution, carrying out trypan exclusion staining, wherein the cell activity is more than 95%, carrying out Wright-Giemsa staining, carrying out morphological observation on 80% neutrophile cells, and carrying out mononuclear cells.

2.2. Leukotriene B₄Generation of

Cells collected as above were adjusted to 5X 10 with Hanks balanced salt buffer⁶and/mL, subpackaging 1.0mL, incubating at 37 ℃ for 10min, sequentially adding L-cysteine (10mM), indometacin (1mg/L) and each test compound, incubating at 37 ℃ for 30min, adding calcium ionophore A23187(5 mu M), incubating at 37 ℃ for 30min, immediately centrifuging at 4 ℃ and 14000r/min for 5min, and storing the supernatant at-70 ℃ for later use, wherein the storage life is not more than 7 days. The final concentration of the solvent in the reaction system is less than or equal to 0.21 percent.

2.3. Leukotriene B₄Measurement of (2)

Diluting the cell extract with buffer solution of commercial EIA kit, adding 96-well enzyme labeling plate, setting two concentrations (0.5 μ M and 5 μ M) for each compound, incubating at 4 deg.C overnight, adding color developing agent the next day, reacting in dark for 90min, detecting absorbance at 412nm, and converting the detection sample according to standard curve established by standard sampleLTB in the product₄The content of (a).

3 statistical analysis

The data were processed using Excell statistical software and each compound produced LTB on neutrophils₄The formula for calculating the inhibition ratio of (a) is:

inhibition rate ═ (concentration in solvent tube-concentration in sample tube)/concentration in solvent tube × 100%

According to the formula, the LTB generated by the compounds with different concentrations on the neutrophils is obtained₄The inhibition ratio of (3).

4 results

4.1 creation of Standard Curve

The logistic regression equations established according to the concentration gradient of the standard are respectively as follows: y ═ 0.1055ln (x) +0.8122, R²＝0.9872；Y＝-0.0487ln(x)+0.4309，R²＝0.978。

4.2 production of test Compounds on neutrophils Release of LTB₄Inhibition rate of

LTB stimulated by calcium ionophore A23187 after addition of varying concentrations of test compound to neutrophil incubation₄Producing various degrees of inhibition. The results of the two concentration prescreening experiments are shown in Table 2, and most of the compounds exhibited inhibitory activity at both 0.5. mu.M and 5. mu.M test concentrations, with 8 of the compounds having greater than 60% inhibitory activity at 5. mu.M test concentration, and the 5 compounds with the better prescreening activity were selected for further IC₅₀The test results are shown in table 2, and most of the compounds showed inhibitory activity comparable to that of marketed drug zileuton, where IC of compounds 1, 3 and 10₅₀Is superior to zileuton. In conclusion, preliminary cell level activity tests show that the compounds are potent 5-LOX inhibitors and have further development and application prospects.

Table 2: flavonoid compound 1-12 isolated rat neutrophil (5X 10) stimulated by calcium ionophore A23187⁶/mL) of LTB₄Released byInhibition of

Industrial applicability

The substituted flavonoid compound disclosed by the invention shows strong 5-LOX inhibitory activity in a 5-LOX cell model test, so that the compound disclosed by the invention can be developed as a 5-LOX inhibitor and used for preparing medicines for preventing and/or treating inflammatory diseases, immune diseases such as asthma and allergy and cardiovascular diseases.

Claims

1. Substituted flavonoid compounds shown in the following general formula (I) and pharmaceutically acceptable salts thereof or solvates or hydrates thereof:

wherein,

x and Y are each independently selected from O, S or NH;

R₂and R₄Each independently selected from hydrogen, trifluoromethyl, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated hydrocarbyl, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated alkanoyl, substituted or unsubstituted C1-C6 linear or branched saturated or unsaturated alkylsulfonyl, substituted or unsubstituted aromatic cyclylsulfonyl or aromatic heterocyclylsulfonyl, substituted or unsubstituted aromatic ring group or aromatic heterocyclic group, substituted or unsubstituted aryl (C1-C4 linear or branched saturated or unsaturated) alkanoyl;

Or- (CH)₂CH ═ CH) -, to form a cyclic structure;

R₆～R₁₀each independently selected from hydrogen, hydroxyl, halogen atom, sulfhydryl, nitro, cyano, amino, trifluoromethyl, hydroxymethyl, carboxyl, C1-C6 linear or branched saturated or unsaturated alkyl, C1-C6 linear or branched saturated or unsaturated alkoxy, and C1-C6 linear or branched saturated or unsaturated alkylamino.

2. Substituted flavonoids according to claim 1And pharmaceutically acceptable salts thereof or solvates or hydrates thereof, characterized in that X is O, Y is O, and R is₁Is 3-methylbut-2-enyl or 5-methylhexa-2, 4-dienyl, and the R radical₂～R₁₀As defined in claim 1.

3. The substituted flavonoid compound and the pharmaceutically acceptable salt thereof or the solvate or hydrate thereof according to claim 1 or 2, wherein the aromatic ring group of the aromatic ring sulfonyl group and the aromatic ring group is phenyl, tolyl, xylyl, biphenyl, naphthyl, indenyl, anthryl or phenanthryl.

4. The substituted flavonoid compound and the pharmaceutically acceptable salt thereof or the solvate or hydrate thereof according to claim 1 or 2, wherein the aromatic heterocyclic group in the aromatic heterocyclic sulfonyl group and the aromatic heterocyclic group is furyl, thienyl, pyrrolyl, imidazolyl, thiazolyl, pyrazolyl, isothiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazolyl or tetrazolyl.

5. Substituted flavonoids according to claims 1 or 2 and their pharmaceutically acceptable salts or solvates or hydrates thereof, characterized in that the aromatic (saturated or unsaturated, linear or branched, C1-C4) alkanoyl is a combination of the aromatic ring group of claim 3 with a saturated or unsaturated, linear or branched, C1-C4 alkanoyl.

6. Substituted flavonoids according to claim 5, and the pharmaceutically acceptable salts or solvates or hydrates thereof, characterized in that the aryl (saturated or unsaturated, linear or branched, C1-C4) alkanoyl is benzoyl, phenylacetyl, phenylpropionyl, phenylbutyryl, (Z) styroyl, p-phenylbenzoyl, p-phenylphenylacetyl, p-phenylphenylpropionyl, p-phenylphenylbutyryl.

7. Substituted flavonoids according to claim 1 or 2 and their pharmaceutically acceptable salts or solvates or hydrates thereof, characterized in that the substituents are one or more groups selected from the group comprising halogen atoms, cyano groups, nitro groups, amino groups, hydroxyl groups, hydroxymethyl groups, trifluoromethyl groups, trifluoromethoxy groups, carboxyl groups, mercapto groups, linear or branched, saturated or unsaturated alkyl groups from C1 to C4, linear or branched, saturated or unsaturated alkoxy groups from C1 to C4, linear or branched, saturated or unsaturated alkanoyl groups from C1 to C4, linear or branched, saturated or unsaturated alkylamino groups from C1 to C4.

8. Substituted flavonoids or their pharmaceutically acceptable salts or their solvates or hydrates according to claim 1, characterized in that the compounds are selected from the following compounds:

1)5, 7-dihydroxy-3- (3-methylbut-2-enyl) -2-phenyl-4H-chromen-4-one;

9. A process for preparing a substituted flavonoid compound or a pharmaceutically acceptable salt thereof or a solvate or hydrate thereof according to claim 1, comprising the steps of:

(a) taking 3-oxo ethyl phenylpropionate as a raw material, and reacting the raw material with a halide R under an alkaline condition₁Performing nucleophilic substitution reaction on X to prepare 2-substituted 3-oxophenpropanoic acid ethyl ester, wherein X is Cl or Br, R₁As defined in claim 1;

(b) 2-substituted 3-oxo ethyl phenylpropionate reacts with phloroglucinol under the microwave condition to prepare a 3-substituted 5, 7-dihydroxy flavone derivative;

(c) condensing the 3-substituted 5, 7-dihydroxyflavone derivative obtained in the step (b) and 3-methyl-2-butenal under microwave or heating conditions;

(d) reacting the 3-substituted 5, 7-dihydroxyflavone derivative obtained in step (b) with an electrophile R₄X or sulfonate R₄OSO₂CH₃Condensing under alkaline condition, wherein X is Cl or Br, R₄As defined in claim 1;

(e) the product obtained in step (c) is optionally further reacted with an electrophile R₂X or sulfonate R₂OSO₂CH₃Condensation reaction under alkaline condition, wherein X is Cl or Br, R₂As defined in claim 1;

(f) if necessary, carrying out salt-forming reaction on the products of the step (d) and the step (e);

wherein the base used in steps (a), (d) and (e) is selected from organic bases including pyridine, triethylamine, 4-dimethylaminopyridine, diisopropylethylamine, potassium tert-butoxide and inorganic bases including sodium carbonate, potassium carbonate, cesium carboxylate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride.

10. A process for preparing a substituted flavonoid compound or a pharmaceutically acceptable salt thereof or a solvate or hydrate thereof according to claim 1, comprising the steps of:

(a) taking 2, 4, 6-trihydroxyacetophenone containing or not containing substituent groups as a raw material, and firstly protecting two hydroxyl groups at 2, 4 positions by MOMCl;

(b) reacting the product of step (a) with R under basic conditions₆-R₁₀Acylation reaction of substituted benzoyl chloride, wherein, the substituent R₆-R₁₀As defined in claim 1, the base used is pyridine;

(c) the product of the step (b) is subjected to rearrangement reaction under the alkaline condition, wherein the used base is selected from organic bases including 4-dimethylamino pyridine, sodium methoxide, sodium ethoxide and potassium tert-butoxide and inorganic bases including sodium hydroxide, potassium hydroxide and sodium hydride;

(d) the product of the step (a) is directly subjected to alkaline condition and then subjected to one-step method to complete acylation and rearrangement reaction, wherein an acylation reagent is R₆-R₁₀Substituted benzoyl chloride, substituent R₆-R₁₀As defined in claim 1, the base used is selected from the group consisting of organic bases including sodium methoxide, sodium ethoxide, potassium tert-butoxide and inorganic bases including sodium hydroxide, potassium hydroxide, sodium hydride;

(e) reacting the product of step (c) or step (d) with a halide R under basic conditions₁X is subjected to nucleophilic substitution reaction, wherein X is Cl or Br, R₁As defined in claim 1;

(f) deprotecting and cyclizing the product obtained in the step (e) under an acidic condition to prepare a 3-substituted 5, 7-dihydroxyflavone derivative;

(g) condensing the 3-substituted 5, 7-dihydroxyflavone derivative obtained in the step (f) and 3-methyl-2-butenal under microwave or heating conditions;

(h) reacting the 3-substituted 5, 7-dihydroxyflavone derivative obtained in the step (f) with an electrophilic reagent R under alkaline conditions₄X or sulfonate R₄OSO₂CH₃Condensation, wherein X is Cl or Br, R₄As defined in claim 1;

(i) the product obtained in step (g) is optionally further reacted with an electrophile R₂X or sulfonate R₂OSO₂CH₃Condensation reaction under alkaline condition, wherein X is Cl or Br, R₂As defined in claim 1;

(j) (ii) if necessary, carrying out a salt-forming reaction on the products of the step (h) and the step (i);

wherein the base used in steps (e), (h) and (i) is selected from organic bases including pyridine, triethylamine, 4-dimethylaminopyridine, diisopropylethylamine, potassium tert-butoxide and inorganic bases including sodium carbonate, potassium carbonate, cesium carboxylate, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride.

11. The production method according to claims 9 and 10, characterized in that the organic solvent used in the synthesis of the target compound (I) is selected from anhydrous inert solvents including diethyl ether, tetrahydrofuran, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, N-methylpyrrolidone, pyridine, dichloromethane, chloroform, N-hexane, benzene, toluene.

12. The substituted flavonoid compound and the pharmaceutically acceptable salt thereof or the solvate or hydrate thereof according to any one of claims 1 to 8 as a 5-lipoxygenase inhibitor for use in the preparation of medicaments for preventing and/or treating inflammatory diseases, immunological diseases such as asthma and allergy, and cardiovascular diseases.

13. A pharmaceutical composition comprising a therapeutically effective amount of a substituted flavonoid compound or a pharmaceutically acceptable salt thereof or a solvate or hydrate thereof according to any one of claims 1 to 8 and at least one pharmaceutically acceptable carrier.