CN104945281A - Flavone acetate derivatives, and pharmaceutical composition, preparation method and application thereof - Google Patents

Abstract

The invention belongs to the field of medical chemical industry, and relates to flavone acetate derivatives, and a pharmaceutical composition, preparation method and application thereof, particularly compounds disclosed as Formula I or Formula II. The compounds have strong actions on inducing tumor cells to generate TNF-alpha and breaking vessels in the chick chorioallantoic membrane, are an effective tumor vessel inhibitor or breaker, and have the prospects in preparing antineoplastic drugs.

Description

Flavone acetic acid analog derivative, its pharmaceutical composition, Preparation Method And The Use

Technical field

The invention belongs to field of medicine and chemical technology, relate to a kind of flavone acetic acid analog derivative, its pharmaceutical composition, Preparation Method And The Use.

Background technology

Tumour needs to rely on functional blood vessel network provide oxygen, nutriment for it and remove meta-bolites in time.Except obtaining part blood vessel by integrating with host blood vessel, tumour also must build oneself vascular system by forming new vessel net could advolution constantly.If do not have vascular system to provide oxygen and nutriment, the growth of solid tumor can not more than 1mm ³.In view of its vital role in tumor development, tumor vessel has become an important target spot of antineoplaston.

Tumor vessel disrupting agent can destroy tumor vascular medicine fast and selectively.The difference that it utilizes tumor vessel and Normal tissue vascular to exist optionally destroys tumor vessel, or by specific combination tumor vascular part toxin, blood coagulation inductor, apoptosis induction molecule etc. being transported to tumor vessel, thus damage fast and selectively or block the tumor vessel built.Form tumour for treatment blood vessel, tumor vessel disrupting agent has significant curative effect.Small molecules tumor vessel disrupting agent can be quick, destroys the tumor vessel formed widely, becomes the focus of research.Comprised colchicine, combretastatin A-4 P, flavone acetic acid class etc. by the tumor vessel disrupting agent generally studied at present, be applied to treatment (the Disrupting Tumour Blood Vessels of the solid tumor such as nonsmall-cell lung cancer, mammary cancer, Nature Reviews, June 2005 Volume 5:423-435).

Flavonoid tumor vessel disrupting agent is the one of small molecules tumor vessel disrupting agent, generally believes that it destroys tumor vascular effect by directly destroying tumor vascular endothelial cell and promoting that tumor tissue cell release tumor necrosis factor α (TNF-α) etc. plays indirectly at present.Flavone acetic acid (FAA) is the flavonoid tumor vessel disrupting agent be found the earliest, it produces TNF-α by inducing tumor cell, the concentration of the TNF-α in remarkable rising tumor microenvironment, the vascular endothelial cell generation apoptosis of tumor by local is played a role, and the general toxic reaction of TNF-α can be avoided.But FAA is only effective in mouse tumor model, does not have antitumous effect in clinical trial.

DMXAA (DMXAA) is the FAA derivative with greater activity, and activity is better than FAA (research finds that FAA is only effective in animal level), but research shows that it only has the reversible toxicity of slight dosage.In addition, in III clinical trial phase, DMXAA fails the survival time of significant prolongation patient.

Still need at present and will develop new antineoplastic compound.

Summary of the invention

The present inventor, through deep research and performing creative labour, obtains a kind of new flavone acetic acid analog derivative.The present inventor is surprised to find, and this compounds is effective tumor vessel disrupting agent, has stronger inducing tumor cell and produces TNF-α effect, and to the destruction of chick chorioallantoic membrane when medium vessels, may be used for preparing antitumor drug.Thus provide following invention:

One aspect of the present invention relates to the compound shown in formula I, or its pharmacologically acceptable salt or ester,

Wherein:

R ₁be selected from H, C ₁-C ₆alkyl;

When the aceticoceptor in female ring is positioned at R ₁during position, R ₁do not exist;

R ₂be selected from the phenyl of phenyl, the replacement of one or more (such as 2,3,4 or 5) hydroxyl, and be selected from following group:

particularly, for R ₂for other substituting group above-mentioned except phenyl ring, R ₂with unsubstituted any site that the link position of female ring is on phenyl ring in R2;

R ₃be selected from H ,-OH.

Another aspect of the present invention relates to the compound shown in formula II, or its pharmacologically acceptable salt or ester,

Wherein:

R ₁be selected from H, C ₁-C ₆alkyl;

When the aceticoceptor in female ring is positioned at R ₁during position, R ₁do not exist;

R ₂be selected from the phenyl of phenyl, the replacement of one or more (such as 2,3,4 or 5) hydroxyl, and be selected from following group:

particularly, for R ₂for other substituting group above-mentioned except phenyl ring, R ₂with unsubstituted any site that the link position of female ring is on phenyl ring in R2;

R ₃be selected from H ,-OH;

represent singly-bound or double bond.

Formula I according to any one of the present invention or formula II compound, or its pharmacologically acceptable salt, wherein,

R ₁independently selected from H, C ₁-C ₃alkyl; Particularly, described C ₁-C ₃alkyl is methyl, ethyl, propyl group or sec.-propyl; Preferably, R ₁be H or methyl independently.

Formula I according to any one of the present invention or formula II compound, or its pharmacologically acceptable salt, wherein,

R ₂independently selected from wherein dotted line represents R ₂with the link position of female ring.

Formula I according to any one of the present invention or formula II compound, or its pharmacologically acceptable salt, wherein, preferably, described formula I or formula II compound do not comprise following 4 compounds:

and

Formula I according to any one of the present invention or formula II compound, wherein, described ester is the ester that the aceticoceptor in female ring is formed; Particularly, be described aceticoceptor and C ₁-C ₆the ester that saturated monohydroxy alcohol (alkanol) is formed; Particularly, described C ₁-C ₆saturated monohydroxy alcohol is methyl alcohol, ethanol, propyl alcohol, Virahol, propyl carbinol, amylalcohol or hexanol.

The present inventor also finds, ester of the present invention is also effective tumor vessel disrupting agent, has stronger inducing tumor cell and produces TNF-α effect, and to the destruction of chick chorioallantoic membrane when medium vessels, may be used for preparing antitumor drug.

Described formula I or formula II compound, or its pharmacologically acceptable salt or ester, wherein, described formula I or formula II compound are selected from table 1 below:

Table 1: part of compounds of the present invention

Another aspect of the invention relates to the preparation method of the formula I according to any one of the present invention, comprises the steps:

(1) preparation of A fragment:

(2) preparation of B2, B3, B4, B5, B6 fragment:

(3) take ethanol as solvent, under the effect of 60% potassium hydroxide aqueous solution, by A fragment respectively with B ₁fragment namely b ₂fragment, B ₃fragment, B ₄fragment, B ₅fragment or B ₆fragment is obtained by reacting formula I;

Wherein, dotted portion represents R ₁.

Preparation method according to any one of the present invention, wherein, described step (1) comprising:

A) with for raw material and paraformaldehyde and concentrated hydrochloric acid are obtained by reacting

B) in organic solvent (such as toluene), under reagent (such as Tetrabutyl amonium bromide) exists, make react with sodium cyanide, obtain further hydrolysis, obtains A fragment (such as ).

Preparation method according to any one of the present invention, wherein, described step (2) comprising:

C) B ₁fragment with B ₂fragment can directly buy;

D) B ₂fragment and bromo isopentene react under solution of potassium carbonate condition, B when obtaining ₃fragment

E) B is made ₃fragment and methoxymethyl chlorine react, and obtain B ₄fragment

F) by B ₄fragment respectively with tosic acid, 2,3-bis-chloro-5,6-dicyanos-benzoquinones, obtain B ₅fragment b ₆fragment

According to detailed teachings of the present invention and existing synthetic chemistry knowledge, those skilled in the art easily can synthesize formula I of the present invention.

Another aspect of the invention relates to the preparation method of the formula II compound according to any one of the present invention, and it comprises the method for the preparation I compound according to any one of the present invention, and comprises:

(4) formula II compound is obtained by formula I; Comprise the steps: particularly

By formula I through DMSO/I ₂flavonoid compound (such as reference example 17) is obtained by reacting under/microwave condition; Flavones alkyl compound (such as reference example 35) is obtained by reacting under TFA/ microwave condition; Formula II compound (such as reference example 53) is obtained by reacting under hydrogen peroxide/NaOH condition.

In one embodiment, the present invention relates to the preparation method preparing the compound shown in general formula I, formula II.Specifically, the invention provides the method preparing the compound shown in general formula I, formula II, comprise the following steps:

1) synthesis of A fragment

As reaction scheme, with o-hydroxyacetophenone starting raw material, itself and paraformaldehyde and concentrated hydrochloric acid are obtained by reacting 2-hydroxyl-3-chloromethyl-methyl phenyl ketone and 2-hydroxyl-5-chloromethyl-methyl phenyl ketone, 2-hydroxyl-3-chloromethyl-methyl phenyl ketone is dissolved in toluene, under Tetrabutyl amonium bromide exists, be obtained by reacting 2-hydroxyl-3-cyano-acetophenone with sodium cyanide, itself and strong sulfuric acid response are obtained A ₁fragment 2-(3-ethanoyl-2-hydroxyphenyl) acetic acid, same method obtains A ₂fragment 2-(3-ethanoyl-4-hydroxyphenyl) acetic acid.

With 2-hydroxy-5-methyl base-methyl phenyl ketone for raw material, working method is the same, obtains A ₃fragment 2-(3-ethanoyl-2-hydroxy-5-methyl phenyl) acetic acid.

2) synthesis of B fragment

As reaction scheme, B ₂fragment and bromo isopentene react in wet chemical, obtain B ₃fragment; Take acetone as solvent, B ₃fragment and methoxymethyl chlorine react, and obtain B ₄fragment; Take toluene as solvent, B ₄fragment is reacted with 2,3-bis-chloro-5,6-dicyanos-benzoquinone, tosic acid respectively, obtains B respectively ₅fragment, B ₆fragment.

C) take ethanol as solvent, under the effect of 60% potassium hydroxide aqueous solution, A ₁fragment, A ₂fragment, A ₃fragment respectively with B ₁fragment, B ₂fragment, B ₃fragment, B ₄fragment, B ₅fragment, B ₆fragment is obtained by reacting formula I, and formula I is by being obtained by reacting formula II compound accordingly.

About preparing general formula I, data that II compound is more detailed is also shown in embodiment.

Another aspect of the invention relates to a kind of pharmaceutical composition, and it comprises formula I according to any one of the present invention or formula II compound or pharmaceutically acceptable salt thereof, and optional pharmaceutically acceptable auxiliary material; Alternatively, it also comprises lipopolysaccharides.

Term " composition " means to comprise the product of each appointment composition comprising specified amount, and any product directly or indirectly produced from the combination of each appointment composition of specified amount.

Usual pharmaceutical composition of the present invention contains the compounds of this invention and/or its pharmacy acceptable salt of 0.1-90 % by weight.Pharmaceutical composition can be prepared according to methods known in the art.During for this object, if needed, the compounds of this invention and/or its pharmacy acceptable salt and one or more solids or liquid pharmaceutical excipients and/or assistant agent can be combined, make the suitable administration form or dosage form that can be used as people.

Compound of the present invention or the pharmaceutical composition containing it can administrations in a unit, and route of administration can be enteron aisle or non-bowel, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritonaeum or rectum etc.Form of administration is tablet, capsule, dripping pill, aerosol, pill, pulvis, solution, suspensoid, emulsion, granule, liposome, transdermal agent, buccal tablet, suppository, lyophilized injectable powder etc. such as.Can be ordinary preparation, sustained release preparation, controlled release preparation and various particulate delivery system.In order to unit dosage forms for administration is made tablet, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as starch, dextrin, calcium sulfate, lactose, N.F,USP MANNITOL, sucrose, sodium-chlor, glucose, urea, calcium carbonate, white bole, Microcrystalline Cellulose, pure aluminium silicate etc.; Wetting agent and tackiness agent, as water, glycerine, polyoxyethylene glycol, ethanol, propyl alcohol, starch slurry, dextrin, syrup, honey, glucose solution, mucialga of arabic gummy, gelatine size, Xylo-Mucine, lac, methylcellulose gum, potassiumphosphate, polyvinylpyrrolidone etc.; Disintegrating agent, such as dry starch, alginates, agar powder, laminaran, sodium bicarbonate and Citric Acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid ester, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; Disintegration inhibitor, such as sucrose, Tristearoylglycerol, theobroma oil, hydrogenation wet goods; Absorption enhancer, such as quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant, such as talcum powder, silicon-dioxide, W-Gum, stearate, boric acid, whiteruss, polyoxyethylene glycol etc.Tablet can also be made coating tablet further, such as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablets and multilayer tablet.In order to administration unit is made pill, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as glucose, lactose, starch, theobroma oil, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talcum powder etc.; Tackiness agent is as gum arabic, tragacanth gum, gelatin, ethanol, honey, liquid sugar, rice paste or batter etc.; Disintegrating agent, as agar powder, dry starch, alginates, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.In order to administration unit is made suppository, various carrier well known in the art can be widely used.Example about carrier is, the ester, gelatin, semi-synthetic glyceryl ester etc. of such as polyoxyethylene glycol, Yelkin TTS, theobroma oil, higher alcohols, higher alcohols.In order to administration unit is made capsule, effective constituent compound or pharmaceutically acceptable salt thereof is mixed with above-mentioned various carriers, and the mixture obtained thus is placed in hard obviously capsule or soft capsule.Also effective constituent compound or pharmaceutically acceptable salt thereof can be made microcapsule, be suspended in aqueous medium and form suspensoid, also can load in hard capsule or make injection application.In order to administration unit is made injection preparation, as solution, emulsion, lyophilized injectable powder and suspensoid, all thinners that this area is conventional can be used, such as, the isooctadecanol of water, ethanol, polyoxyethylene glycol, 1,3-PD, ethoxylation, polyoxygenated isooctadecanol, Polyoxyethylene Sorbitol Fatty Acid Esters etc.In addition, in order to prepare isotonic injection liquid, appropriate sodium-chlor, glucose or glycerine can be added in injection preparation, in addition, conventional solubility promoter, buffer reagent, pH adjusting agent etc. can also be added.

In addition, as needs, also tinting material, sanitas, spices, correctives, sweeting agent or other material can be added in pharmaceutical preparation.

Compound of the present invention, or the dosage of its pharmacologically acceptable salt depends on many factors, such as, to prevent or the character of disease therapy and severity, the sex of patient or animal, age, body weight and individual reaction, particular compound used, route of administration and administration number of times etc.Above-mentioned dosage can single dose form or be divided into several, such as two, three or four dosage forms for administration.

The active compound amount of gained the actual dose level of each activeconstituents in pharmaceutical composition of the present invention can be changed, so that effectively can obtain required therapeutic response for concrete patient, composition and administering mode.Dosage level must according to the activity of particular compound, route of administration, treat the severity of the patient's condition and the patient's condition of patient to be treated and medical history and select.But the way of this area is, the dosage of compound, from lower than for obtaining level that required result for the treatment of requires, increases dosage, gradually until obtain required effect.

Formula I according to any one of another aspect of the invention relates to or formula II compound or pharmaceutically acceptable salt thereof or pharmaceutical composition of the present invention are being prepared as follows the purposes in the medicine described in any one:

1) medicine such as raising TNF & alpha levels is regulated;

2) inducing tumor cell produce tumor necrosis factor alpha medicine '

3) tumor vessel or the angiopoietic medicine of Tumor suppression is destroyed;

4) medicine of inhibition tumor cell or vascular endothelial cell; And

5) treat and/or prevent and/or adjuvant therapy of tumors medicine in purposes;

Particularly, described tumour cell is colon cancer cell (such as DLD-1 human colon cancer cell), RAW 264.7 scavenger cell, breast cancer cell (such as MDA-MB-231 breast cancer cell) or cervical cancer cell (such as Hela cervical cancer cell).

Particularly, described tumour is lung cancer, mammary cancer, cervical cancer or colorectal carcinoma.

Another aspect of the invention relates to a kind of method be selected from described in following any one, comprises and uses the formula I according to any one of the present invention of significant quantity or the step of formula II compound or pharmaceutically acceptable salt thereof or pharmaceutical composition of the present invention:

1) in vivo or external adjustment such as raise the method for TNF & alpha levels;

2) in vivo or external evoked tumour cell produce the method for tumor necrosis factor alpha; And

3) in vivo or extracorporeal suppression tumor cell such as inhibition tumor cell propagation and/or migration method.

In one embodiment of the invention, above-mentioned method is in vitro non-therapeutic purpose.

Another aspect of the invention relates to a kind for the treatment of and/or preventing and/or the method for adjuvant therapy of tumors, comprises and uses the formula I according to any one of the present invention of significant quantity or the step of formula II compound or pharmaceutically acceptable salt thereof or pharmaceutical composition of the present invention; Particularly, described tumour is lung cancer, mammary cancer, cervical cancer, colorectal carcinoma.

Term " significant quantity " refers to the dosage that can realize treating, prevent, alleviate and/or alleviating disease of the present invention or illness in experimenter.

Term " experimenter " can refer to patient or other accept the present composition to treat, to prevent, to alleviate and/or to alleviate the animal of disease of the present invention or illness, particularly Mammals, such as people, dog, monkey, ox, horse etc.

Term " disease and/or illness " refers to a kind of physical state of described experimenter, and this physical state is relevant with disease of the present invention and/or illness.

But it should be understood that total daily dosage portion of the compounds of this invention or pharmaceutical composition must be maked decision within the scope of reliable medical judgment by attending physician.For any concrete patient, concrete treatment effective dose level must be determined according to many factors, and described factor comprises treated obstacle and the severity of this obstacle; The activity of the particular compound adopted; The concrete composition adopted; Age of patient, body weight, general health situation, sex and diet; The administration time of the particular compound adopted, route of administration and excretion rate; The treatment time length; The medicine combinationally using with adopted particular compound or use simultaneously; And the known similar factor of medical field.Such as, the way of this area is, the dosage of compound, from lower than for obtaining level that required result for the treatment of requires, increases dosage, gradually until obtain required effect.In general, compound of the present invention is used for the dosage of Mammals particularly people can between 0.001-1000mg/kg body weight/day, such as, between 0.01-100mg/kg body weight/day, such as, between 0.01-10mg/kg body weight/day.

Various disease of the present invention or illness effectively can be prevented and/or treated according to compound of the present invention.

Another aspect of the invention relates to and is selected from following midbody compound:

and

Another aspect of the invention relates to the arbitrary midbody compound described in the present invention and is preparing antitumor drug or the formula I described in preparation any one of the present invention or purposes in formula II compound.

The beneficial effect of the invention

Compound of the present invention only has stronger inducing tumor cell to produce the effect of TNF-α, and to the destruction of chick chorioallantoic membrane when medium vessels, is effective tumor vessel inhibitor or disrupting agent, has the prospect for the preparation of antitumor drug.In addition, compound of the present invention does not rely on LPS approach and has TNF-α yet and urge secretion.

Accompanying drawing explanation

The knurl body picture of Fig. 1: control (contrast), DMXAA, embodiment 2 compound.Top graduated scale can read tumorous size.

Fig. 2: tumor volume changes.* P<0.05.n=3-5 mouse/group.mean±SEM。

Fig. 3: to angiolysis effect (Hoshest 33342 dyes).Fig. 3 A, control (contrast); Fig. 3 B, DMXAA; Fig. 3 C, embodiment 2 compound.

Embodiment

Below in conjunction with embodiment, embodiment of the present invention are described in detail, but it will be understood to those of skill in the art that the following example only for illustration of the present invention, and should not be considered as limiting scope of the present invention.Unreceipted actual conditions person in embodiment, the condition of conveniently conditioned disjunction manufacturers suggestion is carried out.Agents useful for same or the unreceipted production firm person of instrument, being can by the conventional products of commercial acquisition.

The fusing point of compound is measured by RY-1 melting point apparatus, the non-calibration of thermometer.Mass spectrum is measured by Micromass ZabSpec high resolution mass spectrometer (resolving power 1000). ¹h-NMR is measured by JNM-ECA-400 SUPERCONDUCTING NMR instrument, operating frequency ¹h-NMR 400MHz.

In the context of the present invention, following shortenings is used:

MOM:-methoxymethyl

DMSO: dimethyl sulfoxide (DMSO)

DDQ:2,3-bis-chloro-5,6-dicyanos-benzoquinone

TsOH: tosic acid

Intermediate is prepared below by preparation example 1-7.

the preparation of preparation example 1:2-hydroxyl-3-chloromethyl-methyl phenyl ketone

Reaction process:

Step 1: the preparation of o-hydroxyacetophenone derivative

Under mechanical stirring, o-hydroxyacetophenone 178g (1.30mol), paraformaldehyde 39.5g (1.30mol) and concentrated hydrochloric acid 800mL react 7 hours at 50-60 DEG C.Separate organic phase, aqueous phase, extract aqueous phase with toluene 200mL, with toluene 600mL solubilizing reaction system layer oily matter, merge organic phase, organic phase uses saturated NaHCO successively ₃aqueous solution 300mL × 2 time, saturated aqueous common salt 300mL washs, anhydrous sodium sulfate drying.After filtration, Rotary Evaporators steam solvent, carry out underpressure distillation to residual oil thing, obtaining main distillate fraction has 211.4g, is the mixture of 2-hydroxyl-3-chloromethyl-methyl phenyl ketone and 2-hydroxyl-5-chloromethyl-methyl phenyl ketone.Normal hexane is added wherein, heating for dissolving according to the ratio of 1g ~ 5mL.After cooling, crystallize out is 2-hydroxyl-5-chloromethyl-methyl phenyl ketone 155.1g; After filtering, filtrate refrigeration (4 DEG C) namely has oily matter to separate out, and inclines and supernatant liquor, and place after concentrated about half and can separate out a large amount of crystal, this crystal is 2-hydroxyl-3-chloromethyl-methyl phenyl ketone 38.3g.Total recovery 80.8%.

the preparation of preparation example 2:2-hydroxyl-5-cyano methyl-methyl phenyl ketone

Reaction process:

2-hydroxyl-5-chloromethyl-methyl phenyl ketone 37g (0.2mol) is dissolved in the middle of 500mL toluene (heating dissolution), add Tetrabutyl amonium bromide 3.2g (20mmol), under stirring at room temperature, dropping sodium cyanide 12.5g (0.255mol) is dissolved in the solution in the middle of 80mL water wherein.Within 20 minutes, drip off, 70 DEG C are reacted 1 hour.

Cooling, organic phase is with 500mL × 2 water washing, and organic phase is dry, and filter, concentrating under reduced pressure obtains a large amount of solid.With petrol ether/ethyl acetate (5:1) recrystallization, obtain sterling 31.6g, yield 89.2%.

The preparation of 2-hydroxyl-3-cyano methyl-methyl phenyl ketone is with reference to aforesaid operations.

the preparation of preparation example 3:2-(3-ethanoyl-4-hydroxyphenyl) acetic acid

Reaction process:

2-hydroxyl-5-cyano-acetophenone 8.7g (34.5mmol), adds the sulfuric acid 75mL (dilution of 1:2 volume ratio obtains) of 33%, back flow reaction 1.5 hours.Cooling, adds cold water 100mL, places, separates out a large amount of yellow solid.Filter washing, crude product hot water recrystallization, the hot elimination removal of impurity, filtrate separates out a large amount of faint yellow solid, filters and obtains pure products 8.3g.Yield 87.7%.

The preparation of 2-(3-ethanoyl-2-hydroxyphenyl) acetic acid is with reference to aforesaid operations.

the preparation of preparation example 4:4-hydroxyl-3-(3-methylene radical-2-alkene) phenyl aldehyde

Reaction process:

Salt of wormwood 140g (1mol) is dissolved in 200g water, after heat release terminates, adds para hydroxybenzene aldehyde 61.0g (0.5mol), is stirred to and dissolves completely.Drip bromo isopentene 60.0mL (0.51mol) again, drip off in 5 minutes.React under room temperature, TLC detects 12 hours.

Reaction solution is with the extraction of ether 250mL × 2, and aqueous phase discards; Ether layer merges, and with 5% aqueous sodium carbonate 200mL × 3 washing, aqueous phase discards; Use 0.5N aqueous sodium hydroxide solution 200mL × 3 to wash again, organic phase discards; Aqueous phase is with after concentrated hydrochloric acid acidifying, and with the extraction of ether 200mL × 3, dry, chromatography column is separated, and petrol ether/ethyl acetate (5:1), obtains 14.1g colorless oil, places and slowly solidifies.Yield 14.8%.

the system of preparation example 5:4-(methoxymethoxy)-3-(3-methylene radical-2-alkene) phenyl aldehyde standby

Reaction process:

4-hydroxyl-3-(3-methylene radical-2-alkene) phenyl aldehyde 1.9g (10mmol), be dissolved in the middle of 50mL acetone, add salt of wormwood 2.0g, ice bath adds methoxymethyl chlorine 2.0mL (20mmol) under stirring, within 10 minutes, drip off, then remove ice bath, react 8 hours at normal temperatures.Decompression is spin-dried for solvent, adds ether 30mL, washs respectively with water 20mL, 0.5N sodium hydroxide solution 20mL, saturated aqueous common salt 20mL, dry, filter, chromatography column is separated, petrol ether/ethyl acetate (8:1), obtain colorless oil 2.1g, place and solidify gradually, yield 83.1%.

the preparation of preparation example 6:2,2-dimethyl-2H-chromene-6-formaldehyde

Reaction process:

4-(methoxymethoxy)-3-(3-methylene radical-2-alkene) phenyl aldehyde 1.90g (10mmol) is dissolved in the middle of 30mL toluene, add DDQ (2,3-bis-chloro-5,6-dicyano-benzoquinone) 2.70g (12mmol), reflux 5 hours.Cooled and filtered, filter cake toluene 10mL washs, and filtrate is successively with 15mL water, unsaturated carbonate potassium solution, saturated nacl aqueous solution washing.Organic phase is dry, filters, and chromatography column is separated, and petrol ether/ethyl acetate (10:1), obtains product 0.80g, yield 44.4%.

the preparation of preparation example 7:2,2-diformazan chromene-6-formaldehyde

Reaction process:

4-(methoxymethoxy)-3-(3-methylene radical-2-alkene) phenyl aldehyde 1.90g (10mmol) is dissolved in the middle of toluene 30mL, adds tosic acid 0.20g, refluxes 3 hours.Cooled and filtered, filter cake toluene 10mL washs, and filtrate is successively with 15mL water, unsaturated carbonate potassium solution, saturated nacl aqueous solution washing.Organic phase is dry, filters, and chromatography column is separated, and petrol ether/ethyl acetate (10:1), obtains product 1.58g, yield 83.2%.

embodiment 2:2-(2-hydroxyl-3-(3-(4-hydroxy phenyl) acryl) phenyl) second acid

By 2-(3-ethanoyl-2-hydroxyphenyl) acetic acid 1.00g (5.18mmol) and para hydroxybenzene aldehyde 0.63g (5.18mmol) mixing, add ethanol 6mL and dissolve, add 60% potassium hydroxide aqueous solution 8mL, room temperature reaction 4 hours.Dilute with frozen water 50mL, extract with ether 20mL, ether discards mutually.Under ice bath stirs, aqueous phase hcl acidifying is to pH ≈ 2, and have a large amount of bright yellow solid to separate out, filtration drying, obtains 0.89g product, yield 92.2%.

¹H-NMR(DMSO-d ⁶,400MHz)：3.586(2H，s，-C H ₂-COOH)，6.843-6.981(3H，m，Ar- H×3)，7.498-7.517(1H，dd，Ar- H)，7.806-7.893(4H，m，Ar- H×4),8.253-8.273(1H，m，Ar- H)，10.232(1H，s，-O H)，12.292(1H，s，-O H)，13.492(1H，s，-COO H)。ESI-MS[M+1]:299.1。

embodiment 3:2-(4-hydroxyl-3-(3-(4-(hydroxy phenyl) acryl) phenyl) acetic acid

Synthetic method with reference to embodiment 2 prepares the present embodiment compound, replaces corresponding starting raw material, obtain product 1.45g, yield 94.1% unlike the use of 2-(3-ethanoyl-4-hydroxyphenyl) acetic acid 1.00g (5.18mmol).

¹H-NMR(DMSO-d ⁶,400MHz)：3.603(2H，s，-C H ₂-COOH)，6.778-7.018(3H，m，Ar- H×3),7.326-7.465(2H，m，Ar- H×2)，7.658-7.823(3H，m，Ar- H×3)，8.143-8.148(1H，d，J＝2Hz，Ar- H)，10.231(1H，s，-O H)，12.367(1H，s，-O H)，12.729(1H，s，-COO H)。ESI-MS[M+1]:298.0。

embodiment 4:2-(2-hydroxyl-3-(3-(4-hydroxy phenyl) acryl)-5-methylbenzene base) acetic acid

Synthetic method with reference to embodiment 2 prepares the present embodiment compound, replaces corresponding starting raw material, obtain product 1.43g, yield 88.5% unlike 2-(3-ethanoyl-2-hydroxy-5-methyl phenyl) acetic acid 1.00g (4.83mmol).

¹H-NMR(DMSO-d ⁶,400MHz)：2.325(3H，s，-C H ₃)，3.552(2H，s，-C H ₂-COOH)，6.855-6.877(2H，d，J＝8.8Hz，Ar- H×2),7.336-7.340(1H，d，J＝1.6Hz，Ar- H)，7.818-7.934(4H，m，Ar- H×4),8.093-8.096(1H，d，J＝1.2Hz，Ar- H)，10.235(1H，s，-O H)，12.280(1H，s，-O H)，13.310(1H，s，-COO H)。ESI-MS[M+1]:313.0。

embodiment 9:2-(3-(3-(2,2-dimethyl-6-base) acryl)-2-hydroxy phenyl) acetic acid

Synthetic method with reference to embodiment 2 prepares the present embodiment compound; corresponding starting raw material is replaced unlike the use of 2-(3-ethanoyl-4-hydroxyphenyl) acetic acid 1.65g (8.55mmol) and 4-(methoxymethoxy)-3-(3-methylene radical-2-alkene) phenyl aldehyde 2.00g (8.55mmol); obtain product 1.30g, yield 37.14%.

¹h-NMR (DMSO-d ⁶, 400MHz): 1.688-1.730 (6H, d, J=16.8Hz ,-C h ₃× 2), 3.323 (1H, s ,-C h ₂one of), 3.391 (3H, s ,-C h ₃), 3.618 (2H, s ,-C h ₂-COOH), 5.276-5.317 (3H, m ,-OC h ₂;-C h), 6.943-6.964 (1H, d, J=8.4Hz, Ar- h), 7.111-7.133 (1H, d, J=8.8Hz, Ar- h), 7.444-7.471 (1H, dd, Ar- h), 7.713-7.775 (2H, m, Ar- h× 2), 7.814-7.893 (2H, m, Ar- h× 2), 8.110-8.115 (1H, d, J=2Hz, Ar- h), 12.591 (2H, s ,-O h;-COO h).ESI-MS[M+1]:411.5。

embodiment 10:2-(2-(2-hydroxyl-3-(3-(4-(methoxymethoxy)-3-(3- methyl but-2-ene base) phenyl) acryl)-5-aminomethyl phenyl) acetic acid

Synthetic method with reference to embodiment 2 prepares the present embodiment compound; corresponding starting raw material is replaced unlike the use of 2-(3-ethanoyl-2-hydroxy-5-methyl phenyl) acetic acid 1.77g (8.55mmol) and 4-(methoxymethoxy)-3-(3-methylene radical-2-alkene) phenyl aldehyde 2.00g (8.55mmol); obtain product 1.71g, yield 47.2%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.695-1.741(6H，d，J＝18.4，-C H ₃×2)，2.341(3H，s，-C H ₃)，3.334-3.415(5H，m，-C H ₂-CH；-OCH ₃)，3.562(2H，s，-C H ₂-COOH)，5.287-5.326(3H，m，-OC H ₂；-CH ₂-C H)，7.116-7.139(1H，d，J＝9.2Hz，Ar- H)，7.357-7.360(1H，d，J＝1.2Hz，Ar- H),7.776-7.859(3H，m，Ar- H×3)，7.943-7.982(1H，d，J＝15.6Hz，Ar- H)，8.089(1H，s，Ar- H)，12.296(1H，s，-O H)，13.245(1H，s，-COO H)。ESI-MS[M+1]:425.2。

the synthesis ginseng of embodiment 1,5,6,7,8,11,12,13,14,15,16,68 according to embodiment 2, as table 2 below.

Table 2: with reference to the part of compounds of the 2-in-1 one-tenth of embodiment

embodiment 17:2-(2-(4-hydroxy phenyl)-4-oxo-4H-chromene-8-base) second acid

By 2-(2-hydroxyl-3-(3-(4-hydroxy phenyl) acryl) phenyl) acetic acid 89.4mg (0.3mmol), be dissolved in the middle of 2mL DMSO, add 3mg I ₂, microwave reaction 10 minutes, design temperature 150 DEG C.Extract with ethyl acetate 15mL and water 15mL, organic phase is dry, filters, concentrated, and chromatography column is separated, and methylene chloride/methanol (20:1), obtains yellow solid 67mg, yield 54.4%.

¹H-NMR(DMSO-d ⁶,400MHz)：3.987(2H，s，-C H ₂-COOH)，6.859-6.943(3H，m，Ar- H×3),7.400-7.438(1H，t，J＝7.6Hz，Ar- H)，7.714-7.732(1H，dd，Ar- H)，7.920-7.961(3H，m，Ar- H×3)，10.284(1H，s，-O H)，12.544(1H，s，-COO H).ESI-MS[M+1]:297.1。

embodiment 18:2-(3-hydroxyl-2-(4-hydroxy phenyl)-4-oxo-4H-chromene-8- base) acetic acid

Synthetic method with reference to embodiment 17 prepares the present embodiment compound, unlike the use of 2-(4-hydroxyl-3-(3-(4-hydroxy phenyl) acryl) phenyl) acetic acid 0.3g (1.01mmol)

For starting raw material, obtain product 0.16mg, yield 60.4%.

¹H-NMR(DMSO-d ⁶,400MHz)：3.757(2H，s，-C H ₂-COOH)，6.862-6.955(3H，m，Ar- H×3)，7.699-7.701(2H，d，J＝0.8Hz，Ar- H×2)，7.922-7.975(3H，m，Ar- H×3)，10.318(1H，s，-O H)，12.444(1H，s，-COO H).ESI-MS[M+1]:297.1。

embodiment 20:2-(2-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl)-4-oxo -4H-chromene-8-base) acetic acid

Synthetic method with reference to embodiment 17 prepares the present embodiment compound; be starting raw material unlike the use of 2-(2-hydroxyl-3-(3-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl) acryl) phenyl) acetic acid 0.25g (0.68mmol); obtain product 135mg, yield 54.4%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.719(6H，s，-C H ₃×2)，3.981(2H，s，-C H ₂-COOH)，5.340-5.378(1H，t，J＝7.6Hz，-CH ₂-C H)，6.851-6.951(2H，m，Ar- H×2)，7.402-7.439(1H，t，J＝7.2Hz，Ar- H)，7.711-7.804(3H，m，Ar- H×3),7.933-7.956(1H，dd，Ar- H)，10.266(1H，s，-O H)，12.585(1H，s，-COO H).ESI-MS[M+1]:365.2。

embodiment 22:2-(2-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl)-6-methyl -4-oxo-4H-chromene-8-base) acetic acid

Synthetic method with reference to embodiment 17 prepares the present embodiment compound; be starting raw material unlike the use of 2-(2-hydroxyl-3-(3-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl) acryl)-5-aminomethyl phenyl) acetic acid 0.26g (0.68mmol); obtain product 134mg, yield 51.8%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.715(6H，S，-C H ₃×2)，2.410(3H，S，Ar-C H ₃)，3.335(2H，S，-C H ₂-CH)，3.929(2H，S，-C H ₂-COOH)，5.350-5.356(1H，t，J＝1.2Hz，-CH ₂-C H)，6.808-6.943(2H，m，Ar- H×2)，7.538-7.543(1H，d，J＝2Hz，Ar- H)，7.737-7.779(3H，m，Ar- H×3),10.245(1H，s，-O H)，12.580(1H，s，-COO H).ESI-MS[M+1]:379.1。

embodiment 26:2-(2', 2'-dimethyl-4-oxo-2'H, 4H-2,6'-chromene-8-base) second acid

Synthetic method with reference to embodiment 17 prepares the present embodiment compound; unlike the use of 2-(3-(3-(2; 2-dimethyl-2H-chromene-6-base)-2-acryl)-2-hydroxybenzene) acetic acid 110mg (0.3mmol) is starting raw material; obtain product 54.4mg, yield 49.8%.

¹h-NMR (DMSO-d ⁶, 400MHz): 1.318-2.222 (6H, m ,-C h ₃× 2), 2.852-2.918 (1H, m ,-C h ₂one of), 3.056-3.091 (1H, m ,-C h ₂one of), 3.230-3.319 (3H, m ,-C h;-C h ₂), 4.003 (2H, s ,-C h ₂-COOH), 6.891-6.927 (2H, d, J=5.2Hz, Ar- h× 2), 7.431-7.450 (1H, t, J=7.6Hz, Ar- h), 7.892-7.961 (4H, m, Ar- h× 4), 12.646 (1H, s ,-COO h) .ESI-MS [M+1]: 365.1.

the synthesis of embodiment 19,21,23,24,25,27,28,29,30,31 is with reference to real execute example 17, as table 3 below.

Table 3: the part of compounds of synthesizing with reference to embodiment 17

embodiment 35:2-(2-(4-hydroxy phenyl)-4-oxo chroman-8-base) second acid

By 2-(2-hydroxyl-3-(3-4-hydroxy phenyl) acryl) phenyl) acetic acid 0.25g (0.84mmol), be dissolved in the middle of 2mL trifluoroacetic acid, microwave reaction 10 minutes, design temperature 80 DEG C.After ethyl acetate 20mL dilution, with the washing of water 15mL × 2, organic phase is dry, filtering and concentrating, and chromatography column is separated, and methylene chloride/methanol (35:1), obtains yellow solid 0.11g, yield 44%.

¹h-NMR (DMSO-d ⁶, 400MHz): 2.796-2.846 (1H, dd ,-C h ₂one of), 3.120-3.195 (1H, m ,-C h ₂one of), 3.580 (1H, s ,-C h ₂-COOH), 5.482-5.521 (1H, s ,-C h), 6.772-6.801 (2H, m, Ar- h× 2), 7.016-7.054 (1H, t, J=7.6Hz, Ar- h), 7.305-7.327 (2H, d, J=8.6Hz, Ar- h), 7.506-7.529 (1H, dd, Ar- h), 7.690-7.715 (1H, dd, Ar- h), 9.579 (1H, s ,-O h), 12.335 (1H, s ,-COO h) ESI-MS [M+1]: 299.1.

embodiment 38:2-(2-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl)-4-oxo chroman-8-base) acetic acid

Synthetic method with reference to embodiment 35 prepares the present embodiment compound; be starting raw material unlike the use of 2-(2-hydroxyl-3-(3-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl) acryl) phenyl) acetic acid 120mg (0.33mmol); obtain product 83mg, yield 69.2%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.284(6H，s，-C H ₃×2)，1.758-1.791(2H，t，J＝6.8Hz，-C H ₂-CH)，2.811-2.853(2H，dd，-C H ₂)，3.118-3.193(1H，m，-C H ₂-CH)，3.586(2H，s，-C H ₂-COOH)，5.475-5.507(1H，dd，-O H)，6.722-6.743(1H，d，J＝8.4Hz，Ar- H)，7.019-7.057(1H，m，Ar- H)，7.189-7.232(2H，m，Ar- H×2)，7.509-7.527(1H，dd，Ar- H)，7.694-7.713(1H，dd，Ar- H)，12.276(1H，s，-COO H).ESI-MS[M+1]:367.1。

embodiment 39:2-(2-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl)-4-oxo chroman-6-base) acetic acid

Synthetic method with reference to embodiment 35 prepares the present embodiment compound; be starting raw material unlike the use of 2-(4-hydroxyl-3-(3-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl) acryl) phenyl) acetic acid 120mg (0.33mmol); obtain product 81mg, yield 67.8%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.283(6H，s，-C H ₃×2)，1.756-1.988(2H，t，J＝6.8Hz，-C H ₂-CH)，2.739-2.755(3H，m，-C H ₂，-C H)，3.227-3.316(1H，m，-CH ₂-C H)，3.592(2H，s，-C H ₂-COOH)，5.484-5.516(1H，dd，-O H)，6.727-6.748(1H，d，J＝8.4Hz，Ar- H)，7.001-7.023(1H，d，J＝8.8Hz，Ar- H)，7.215-7.264(2H，m，Ar- H×2)，7.447-7.474(1H，dd，Ar- H)，7.669-7.674(1H，d，J＝2Hz，Ar- H).ESI-MS[M+1]:367.2。

embodiment 40:2-(2-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl-6-methyl-4- oxo chroman-8-base) acetic acid

Synthetic method with reference to embodiment 35 prepares the present embodiment compound; be starting raw material unlike the use of 2-(2-hydroxyl-3-(3-(4-hydroxyl-3-(3-methyl but-2-ene base) phenyl) acryl)-5-aminomethyl phenyl) acetic acid 125mg (0.33mmol); obtain product 84.2mg, yield 67.2%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.173(6H，s，-C H ₃×2)，1.753-1.786(2H，t，J＝6.8Hz，-C H ₂-CH)，2.273(3H，S，Ar-C H ₃)，2.743-2.820(3H，m，-C H，-C H ₂)，3.081-3.156(1H，m，-CH ₂-C H)，5.419-5.452(1H，dd，Ar-O H)，6.715-6.736(1H，d，J＝8.4Hz，Ar- H)，7.176-7.222(2H，m，Ar- H×2),7.341-7.346(1H，d，J＝2Hz，Ar- H)，7.497-7.500(1H，d，J＝1.2Hz，Ar- H)，12.311(1H，s，-COO H).ESI-MS[M+1]:381.2。

the synthesis of embodiment 36,37,44,45,46,47,48,49 is with reference to embodiment 35 synthesis, as table 4 below.

Table 4: the part of compounds of synthesizing with reference to embodiment 35:

embodiment 41:2-(2-(4-(methoxymethoxy)-3-(3-methyl but-2-ene base) phenyl-4-oxo chroman-8-base) acetic acid

2-(2-hydroxyl-3-(3-(4-(methoxymethoxy)-3-(3-methyl but-2-ene base) phenyl) acryl) phenyl) acetic acid 123mg (0.3mmol); be dissolved in the middle of 6mL ethanol; add sodium acetate 180mg, instill several dripping and sodium acetate is dissolved completely.Back flow reaction 2 days.Add ethyl acetate 20mL dilution, with the washing of water 15mL × 2, organic phase is dry, filtering and concentrating, and chromatography column is separated, and methylene chloride/methanol (35:1), obtains yellow solid 33mg, yield 26.8%.

¹h-NMR (DMSO-d ⁶, 400MHz): 1.684-1.690 (6H, d, J=2.4Hz ,-C h ₃× 2), 2.886-2.893 (1H, dd ,-C h ₂one of), 3.105-3.137 (1H, m ,-C h ₂one of), 3.283-3.383 (7H, m ,-C h ₂× 3 ,-C h), 3.599 (2H, s ,-C h ₂-COOH), 5.244-5.272 (3H, t, J=6.4Hz ,-OC h ₃), 5.537-5.569 (1H, dd ,-CH ₂-C h), 7.044-7.069 (2H, t, J=7.6Hz, Ar- h× 2), 7.276-7.296 (2H, t, J=8.0Hz, Ar- h× 2), 7.518-7.537 (1H, d, J=7.6Hz, Ar- h), 7.692-7.711 (1H, d, J=7.6Hz, Ar- h), 12.329 (1H, s ,-COO h) .ESI-MS [M+1]: 411.1.

embodiment 42:2-(2-(4-(methoxymethoxy)-3-(3-methyl but-2-ene base) phenyl-4-oxo chroman-6-base) acetic acid

Synthetic method with reference to embodiment 41 prepares the present embodiment compound; be starting raw material unlike the use of 2-(4-hydroxyl-3-(3-(4-(methoxymethoxy)-3-(3-methyl but-2-ene base) phenyl) acryl) phenyl) acetic acid 123mg (0.3mmol); obtain product 31mg, yield 25.2%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.683(6H，s，-C H ₃×2)，2.746-2.795(1H，dd，-CH ₂-C H)，3.286-3.380(6H，m，-C H ₂×3)，3.591(2H，S，-C H ₂-COOH)，5.237-5.256(3H，S，-C H ₃)，5.542-5.567(1H，dd，-CH ₂-C H)，7.008-7.029(2H，m，Ar- H×2)，7.305-7.318(2H，m，Ar- H×2)，7.451-7.478(1H，dd，Ar- H)，7.664-7.669(1H，d，J＝2Hz，Ar- H)，12.291(1H，s，-COO H).ESI-MS[M+1]:411.1。

the synthesis of the synthesis reference embodiment 41 of embodiment 43.

embodiment 53:2-(3-hydroxyl-2-(4-hydroxy phenyl)-4-oxo-4H-chromene-8- base) acetic acid

2-(2-hydroxyl-3-(3-(4-hydroxy phenyl) acryl) phenyl) acetic acid 0.25g (0.84mmol), is dissolved in the middle of 6mL methyl alcohol.Under stirring at room temperature, add 16% aqueous sodium hydroxide solution (being about 5N) 1.2mL, system color is red by xanthochromia.Add 30% hydrogen peroxide 0.15mL again, system color is taken off gradually, reacts 1 hour, and the display of ultraviolet 365nM place is strong to be absorbed.Dilute with frozen water 20mL, be then acidified to pH 2, have a large amount of bright yellow solid to separate out, filter, obtain product 0.18g, yield 69.2%.

¹H-NMR(DMSO-d ⁶,400MHz)：3.989(2H，s，-CH ₂-COOH),6.924-6.947(2H，m，Ar- H×2)，7.387-7.426(1H，t，Ar- H)，7.696-7.717(1H，dd，Ar- H)，8.012-8.094(3H，m，Ar- H×3),9.399(1H，s，-O H)，10.126(1H，s，-O H)，12.632(1H，s，-COO H)。ESI-MS[M+1]:313.1。

embodiment 54:2-(3-hydroxyl-2-(4-hydroxy phenyl)-4-oxo-4H-chromene-6- base) acetic acid

Synthetic method with reference to embodiment 53 prepares the present embodiment compound; be starting raw material unlike the use of 2-(4-hydroxyl-3-(3-(4-hydroxy phenyl) acryl) phenyl) acetic acid 200mg (0.67mmol); obtain product 0.16g, yield 76.2%.

¹H-NMR(DMSO-d ⁶,400MHz)：3.771(2H，s，-C H ₂-COOH)，6.936-6.958(2H，d，J＝2Hz，Ar- H×2)，7.671-7.681(2H，t，J＝2Hz，Ar- H×2)，8.089-8.120(3H，d，J＝8.4Hz，Ar- H×3)，9.337(1H，s，Ar-O H)，10.084(1H，s，-O H)，12.433(1H，s，-COO H)。ESI-MS[M+1]:313.0。

embodiment 59:2-(3-hydroxyl-2-(4-(methoxymethoxy)-3-(3-methyl fourth-2- thiazolinyl) phenyl-4-oxo-4H-chroman-8-base) acetic acid

Synthetic method with reference to embodiment 53 prepares the present embodiment compound; be starting raw material unlike the use of 2-(2-hydroxyl-3-(3-(4-(methoxymethoxy)-3-(3-methyl but-2-ene base) phenyl) acryl) phenyl) acetic acid 200mg (0.5mmol); obtain product 133.7mg, productive rate 64.7%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.717-1.738(6H，d，J＝8.4Hz，-C H ₃×2)，3.350-3.414(5H，m，-C H ₂-CH；-OC H ₃)，3.990(2H，s，-C H ₂-COOH)，5.324-5.341(3H，m，-OC H ₂；-CH ₂-C H)，7.193-7.215(1H，d，J＝8.8Hz，Ar- H)，7.397-7.435(1H，t，J＝7.6Hz，Ar- H)，7.708-7.730(1H，dd,Ar- H)，8.019-8.061(3H，m，Ar- H×3)，9.491(1H，s，-O H)，12.617(1H，s，-COO H)。ESI-MS[M+1]:425.1。

embodiment 60:2-(3-hydroxyl-2-(4-(methoxymethoxy)-3-(3-methyl fourth-2- thiazolinyl) phenyl)-4-oxo-4H-chromene-6-base) acetic acid

Synthetic method with reference to embodiment 53 prepares the present embodiment compound; be starting raw material unlike the use of 2-(4-hydroxyl-3-(3-(4-(methoxymethoxy)-3-(3-methyl but-2-ene base) phenyl) acryl) phenyl) acetic acid 200mg (0.5mmol); obtain product 132.9mg, yield 64.3%.

¹H-NMR(DMSO-d ⁶,400MHz)：1.707-1.733(6H，d，J＝10.4Hz，-C H ₃×2)，3.314-3.410(5H，m，-C H ₂×2，-C H)，3.773(2H，s，-C H ₂-COOH)，5.275-5.326(3H，s，-OC H ₃)，7.198-7.221(1H，d，Ar- H)，7.686(2H，s，Ar- H×2)，7.991-8.043(3H，m，Ar- H×3)，9.404(1H，s，-O H)，12.366(1H，s，-COO H).ESI-MS[M+1]:425.1。

embodiment 50,51,52,55,56,57,58,61,62,63,64,65,66, the synthesis reference embodiment 53 of 67, as table 5 below.

Table 5: the part of compounds of synthesizing with reference to embodiment 53

Pharmacology test and activity data

As test example 1-3 below.

test example 1:AlphaLISA method detects TNF-α in RAW 264.7 cells and supernatant concentration

Experimental technique is as follows: RAW 264.7 cell in vegetative period of taking the logarithm, be mixed with cell suspension with the RPMI-1640 substratum containing 10% foetal calf serum, inoculate in 96 well culture plates, every hole 100 μ l (containing 2500 RAW264.7 cells), be placed in 37 DEG C, 5%CO ₂in incubator, after adding target compound effect certain hour, by centrifugal for 96 orifice plates 2000r, 10min, 150 μ l supernatants are got in every hole afterwards, are dispensed in three EP, often prop up 50 μ l, frozen for subsequent use in-20 DEG C.Subsequently according to the explanation of AlphaLISA TNF-α test kit, preparation typical curve solution, and get 5 μ l supernatants and be inoculated in White OptiPlate-384 orifice plate, then every hole adds AlphaLISA Anti-Analyte Acceptor beads (final concentration 10 μ g/ml) and Biotinylated Antibody Anti-Analyte (final concentration 1nM) mixed solution 20 μ l (Acdeptor beads:Biotinylated Antibody Anti-Analyte:1X buffer=1:2:198), 23 DEG C hatch 60min after add Donor beads (final concentration is 40 μ g/ml) 25 μ l (Donor beads:1X buffer=1:61.5), EnVision is utilized to detect after 23 DEG C of lucifuges hatch 30min.

Experimental result is as table 6.

test example 2:MTS method measures the cytotoxicity of compound

Experimental technique is as follows: mouse monokaryon scavenger cell strain RAW 264.7 cell in vegetative period of taking the logarithm, cell suspension is mixed with the RPMI-1640 substratum containing 10% foetal calf serum, be inoculated in 96 well culture plates, every hole 100 μ l (containing 2500 tumour cells), be placed in 37 DEG C, 5%CO ₂in incubator.After acting on certain hour after adding target compound, every hole adds MTS solution 10 μ l, hatches 4 hours for 37 DEG C.Its light absorption value is measured at 490nm wavelength place with enzyme-linked immunosorbent assay instrument.

Cell survival rate (%)=(administration group OD-blank group OD)/control group OD × 100%

Experimental result is as table 6.

Table 6: part of compounds activity experiment result

"-" expression detects.

Target compound concentration is 30 μ g/ml, and LPS concentration is 2.5ng/ml, with DMSO or DMSO and LPS for contrast; " a " represents that target compound independent role promotes the secretion rate of RAW 264.7 emiocytosis TNF-α; When " b " expression is share with LPS (lipopolysaccharides 2.5ng/ml), target compound and LPS share the secretion rate promoting RAW 264.7 emiocytosis TNF-α; " c " represents the growth-inhibiting effect of target compound independent role to RAW 264.7 cell; " d " represents that target compound and LPS share the growth-inhibiting effect to RAW 264. cell.

Result shows, and formula I, formula II compound have anti-tumor activity, and therefore formula I, formula II compound can be used as antitumor drug in antineoplastic research, has the potentiality that exploitation becomes antitumor drug.

In addition, general short TNF-α secretion is all by LPS approach, and experimental result shows, and compound of the present invention does not rely on LPS approach and can produce TNF-α yet and urge secretion.

test example 3: the impact of compound on vascular investigated by chick chorioallantoic membrane model

Experimental technique is as follows: aseptically, 10 μ l respective concentration solution title compound are spread evenly across on Whatman filter paper, the 0.3%DMSO solution of negative control coating same volume, fertilization kind of egg is placed in 37 DEG C, the constant-temperature incubation case of 60% relative humidity hatches, after 6 days, the CAM film of Dan Pei lower floor is leaked cruelly.Load sample filter paper is placed in CAM and yolk cyst membrane place blood vessel comparatively multiple location, then uses sterile transparent rubber seal mouth, continue to be cultured to 8 days, remove the transparent rubberized fabric covered on chicken embryo.Medicine-feeding part is taken, process picture, statistics administration group and control group vessel area, calculate the per-cent (employing Adobe Photoshop CS5, Image-Pro Plus 6.0 software carry out visual and follow-up area quantitative to chick chorioallantoic membrane blood vessel, n=2) that two groups of vessel area account for the CAM total area.

Angiolysis rate (Distrupting Ratio, %)=[1-(experimental group vessel area/solvent control group vessel area)] × 100%.

Experimental result is as table 7.

Table 7: target compound is on the impact of chick chorioallantoic membrane blood vessel

The concentration of target compound is 10 μ g/ chicken embryos, take DMSO as contrast.

Experimental result shows, and compound of the present invention can promote tumor cell secretion TNF-α, can also reduce chick chorioallantoic membrane number of blood vessel significantly simultaneously.Therefore, compound of the present invention can be used in preparing antitumor drug.

test example 4: to the tumor vascular destruction of lung cancer tumor-bearing mice

Experimental procedure:

(1) foundation of mouse LLC subcutaneous lotus knurl model

By the C of Lewis lung cancer cell lotus knurl in laboratory ₅₇bL/6 mouse takes off neck and puts to death, soaking disinfection 1 ~ 2min in 75% alcohol.In aseptic operating platform, be separated its subcutaneous tumors block afterwards, normal saline flushing 2 times, 150 ~ 200 order stainless steel sifts be placed in advance through high-temperature sterilization are online, and scissors shreds rear abundant grinding.The underlying glass culture dish filling appropriate stroke-physiological saline solution of screen cloth collects the cell after grinding, and is incorporated in centrifuge tube by all cells liquid collected, the centrifugal 5min of 1500rpm.Abandon supernatant, make cell suspension with appropriate stroke-physiological saline solution is resuspended, fully piping and druming evenly, is transferred to 25cm ²in culturing bottle, sealed membrane sealing is stand-by.

With 75% cotton ball soaked in alcohol cleaning disinfection C ₅₇bL/6 right side of mice armpit skin, it is subcutaneous that 1ml syringe draws the cell suspension injection mouse oxter prepared, and every 0.2ml (every mouse all notes abundant mixing when drawing enchylema), overturns syringe needle after stopping pin 2 ~ 3s and pull out pin.After inoculation, record body weight change and the knurl bulk-growth situation of each group of mouse every other day.By formula V=D (major diameter) × d ²(minor axis)/2 calculate tumor volume.If animal occurs dead, record its survival time and optionally its in-vivo tumour of anatomic observation infects situation.

(2) administration and detection

When Tumor diameter grows to about 10mm, remove into knurl animal.Remain all one-tenth knurl mouse suitably to adjust by tumorous size, make often to organize sample and to try one's best evenly distribute.The corresponding abdominal injection DMXAA (20mg/kg) of reagent group, embodiment 2 compound (20mg/kg), the 1%DMSO of control group abdominal injection respective volume.All mouse are tail vein injection fluorescence dye Hoechst 33342 (800 μ g/mouse, 0.1ml) after administration 3h.Mouse taken off neck after injection 1min to put to death, peel off knurl body and take pictures, weigh, calculate tumor volume, and rapid masking foil is wrapped, frozenly in liquid nitrogen, carry out freezing microtome section, observe under inverted fluorescence microscope.

Experimental result: as shown in accompanying drawing 1,2,3 (3A, 3B, 3C).

Result shows, embodiment 2 compound can destroy tumor vessel, reduces tumor blood perfusion.

Although the specific embodiment of the present invention has obtained detailed description, it will be understood to those of skill in the art that.According to disclosed all instructions, can carry out various amendment and replacement to those details, these change all within protection scope of the present invention.Four corner of the present invention is provided by claims and any equivalent thereof.

Claims (14)

1. the compound shown in formula I, or its pharmacologically acceptable salt or ester,

Wherein:

R ₁be selected from H, C ₁-C ₆alkyl;

When the aceticoceptor in female ring is positioned at R ₁during position, R ₁do not exist;

R ₂the phenyl be selected from phenyl, being replaced by one or more (such as 2,3,4 or 5) hydroxyl, and be selected from following group:

particularly, for R ₂for other substituting group above-mentioned except phenyl ring, R ₂with unsubstituted any site that the link position of female ring is on phenyl ring in R2;

R ₃be selected from H ,-OH.

2. the compound shown in formula II, or its pharmacologically acceptable salt or ester,

Wherein:

R ₁be selected from H, C ₁-C ₆alkyl;

When the aceticoceptor in female ring is positioned at R ₁during position, R ₁do not exist;

R ₂be selected from phenyl, one or more (such as 2,3,4 or 5) hydroxyl replace phenyl, and and be selected from following group:

particularly, for R ₂for other substituting group above-mentioned except phenyl ring, R ₂with unsubstituted any site that the link position of female ring is on phenyl ring in R2;

R ₃be selected from H ,-OH;

represent singly-bound or double bond.

3. formula I according to claim 1 or formula II compound according to claim 2, or its pharmacologically acceptable salt or ester, it meets any one in the item of following (1)-(4) or multinomial,

(1) R ₁independently selected from H, C ₁-C ₃alkyl; Particularly, described C ₁-C ₃alkyl is methyl, ethyl, propyl group or sec.-propyl; Preferably, R ₁be H or methyl independently;

(2) R ₂independently selected from wherein dotted line represents R ₂with the link position of female ring;

(3) described formula I or formula II compound do not comprise following 4 compounds:

and

(4) described ester is the ester that the aceticoceptor in female ring is formed; Particularly, be described aceticoceptor and C ₁-C ₆the ester that saturated monohydroxy alcohol is formed; Particularly, described C ₁-C ₆saturated monohydroxy alcohol is methyl alcohol, ethanol, propyl alcohol, Virahol, propyl carbinol, amylalcohol or hexanol.

4. formula I according to claim 1, or its pharmacologically acceptable salt or ester, wherein, described formula I is selected from:

5. formula II compound according to claim 1, or its pharmacologically acceptable salt or ester, wherein, described formula II compound is selected from:

6. the preparation method of the formula I according to any one of claim 1,3 and 4, comprises the steps:

(1) preparation of A fragment:

(2) preparation of B2, B3, B4, B5, B6 fragment:

(3) take ethanol as solvent, under the effect of 60% potassium hydroxide aqueous solution, by A fragment respectively with B ₁fragment namely b ₂fragment, B ₃fragment, B ₄fragment, B ₅fragment or B ₆fragment is obtained by reacting formula I;

Wherein, dotted portion represents R ₁.

7. method according to claim 6, wherein, described step (1) comprising:

A) with for raw material and paraformaldehyde and concentrated hydrochloric acid are obtained by reacting

B) in organic solvent (such as toluene), under reagent (such as Tetrabutyl amonium bromide) exists, make react with sodium cyanide, obtain further hydrolysis, obtains A fragment (such as or ).

8. method according to claim 6, wherein, described step (2) comprising:

C) B ₁fragment with B ₂fragment can directly buy;

D) B ₂fragment and bromo isopentene react under solution of potassium carbonate condition, B when obtaining ₃fragment

E) B is made ₃fragment and methoxymethyl chlorine react, and obtain B ₄fragment

F) by B ₄fragment respectively with tosic acid, 2,3-bis-chloro-5,6-dicyanos-benzoquinones, obtain B ₅fragment b ₆fragment

9. the preparation method of the formula II compound according to any one of claim 2,3 and 5, it comprises the method for the preparation I compound according to any one of claim 6 to 8, and comprises:

(4) formula II compound is obtained by formula I; Particularly, comprise the steps:

By formula I through DMSO/I ₂flavonoid compound is obtained by reacting under/microwave condition; Flavones alkyl compound is obtained by reacting under TFA/ microwave condition; Formula II compound is obtained by reacting under hydrogen peroxide/NaOH condition.

10. a pharmaceutical composition, it comprises formula I according to any one of claim 1 to 5 or formula II compound or pharmaceutically acceptable salt thereof, and optional pharmaceutically acceptable auxiliary material; Alternatively, it also comprises lipopolysaccharides.

Formula I according to any one of 11. claims 1 to 5 or formula II compound or pharmaceutically acceptable salt thereof or pharmaceutical composition according to claim 10 are being prepared as follows the purposes in the medicine described in any one:

1) medicine such as raising TNF & alpha levels is regulated;

2) inducing tumor cell produce tumor necrosis factor alpha medicine '

3) tumor vessel or the angiopoietic medicine of Tumor suppression is destroyed;

4) medicine of inhibition tumor cell or vascular endothelial cell; And

5) treat and/or prevent and/or adjuvant therapy of tumors medicine in purposes;

Particularly, described tumour cell is colon cancer cell (such as DLD-1 human colon cancer cell), RAW 264.7 scavenger cell, breast cancer cell (such as MDA-MB-231 breast cancer cell) or cervical cancer cell (such as Hela cervical cancer cell);

Particularly, described tumour is lung cancer, mammary cancer, cervical cancer or colorectal carcinoma.

12. 1 kinds of methods be selected from described in following any one, comprise and use the formula I according to any one of claim 1 to 5 of significant quantity or the step of formula II compound or pharmaceutically acceptable salt thereof or pharmaceutical composition according to claim 10:

1) in vivo or external adjustment such as raise the method for TNF & alpha levels;

2) in vivo or external evoked tumour cell produce the method for tumor necrosis factor alpha; And

3) in vivo or extracorporeal suppression tumor cell such as inhibition tumor cell propagation and/or migration method.

13. are selected from following midbody compound:

and

14. arbitrary midbody compounds according to claim 13 are preparing antitumor drug or the purposes in preparation formula I according to claim 1 or formula II compound according to claim 2.