CN103288805B - Pyrimidines containing benzofuran and its production and use - Google Patents

Abstract

The invention discloses and there is pyrimidines and the pharmaceutically acceptable salt thereof that a class of structure shown in formula I contains benzofuran.The invention also discloses the preparation method of above-claimed cpd, and also disclose using this compound or its pharmaceutically acceptable salt as the pharmaceutical composition of active ingredients, and they are as antitumor drug, particularly in the application in terms of preparing treatment breast carcinoma, pulmonary carcinoma, gastric cancer medicament.

Description

Pyrimidines containing benzofuran and its production and use

Technical field

The invention belongs to pharmaceutical technology field, relate to class compound with antitumor action and preparation method thereof and use On the way, more precisely, relate to a class there is the pyrimidines containing benzofuran and the preparation side thereof of antitumor action Method and purposes.

Background technology

The health and lives of the mankind in cancer serious harm.Suffer from cancered people the most every year and have 9,000,000, extremely Patient in cancer is 6,000,000, almost just has a cancer death each second.The M & M of China's cancer occupies Height not under, year, number of the infected was about 1,200,000, and the number dying from cancer is up to more than 900,000, and patient to be treated is more than 150 Ten thousand, and have the trend risen year by year.At present, cancer has become the second largest killer being only second to cardiovascular disease.Control clinically Treat cancer, typically use operation, radiotherapy, the big therapy of chemotherapy three.Though embolic chemotherapy is more quick, but cure rate is the lowest.Face simultaneously Bed finds that many cancer therapy drugs exist the obvious damage to normal body and toxic and side effects, such as mutagenesis and genetoxic. Therefore, find effective and that there is less body injury and toxic and side effects cancer therapy drug and become the focus of new drug research.

Summary of the invention

It is an object of the present invention to disclose the pyrimidines containing benzofuran of a class new structure and Pharmaceutical salts.

Further object is that, disclose pyrimidines that a class contains benzofuran and pharmaceutical salts thereof Preparation method.

Another object of the present invention is, open pyrimidines and the pharmaceutical salts thereof containing benzofuran with a class Pharmaceutical composition for main active.

A further object of the invention is, discloses pyrimidines and pharmaceutical salts work thereof that a class contains benzofuran For the application of medicine for resisting malignant tumors, particularly in the use in terms of preparing treatment breast carcinoma, pulmonary carcinoma, gastric cancer medicament On the way.

In conjunction with the object of the invention, present invention is described in detail.

Present invention relates particularly to compound and the pharmaceutically acceptable salt thereof of structure shown in formula I:

Wherein:

R₁For: hydrogen, halogen, alkyl, alkoxyl；

R₂For hydrogen, alkyl, phenyl, methyl substituted-phenyl or halogen substituted phenyl orWherein R₃For phenyl, Methyl substituted-phenyl, halogen substituted phenyl, saturated or unsaturated quinary heterocyclic radical.

The preferred following compound of compound of structure shown in formula I and pharmaceutically acceptable salt thereof:

Wherein:

R₁For: hydrogen, fluorine, chlorine, bromine, methyl, ethyl, methoxyl group；

R₂For hydrogen, methyl, ethyl, phenyl, methyl substituted-phenyl or halogen substituted phenyl orWherein R₃For Phenyl, methyl substituted-phenyl, halogen substituted phenyl, saturated or unsaturated quinary heterocyclic radical.

The following compound of compound of structure shown in formula I and pharmaceutically acceptable salt thereof:

Wherein:

R₁For: hydrogen, methyl, fluorine, methoxyl group；

R₂For hydrogen, methyl, phenyl orWherein R₃For: phenyl, furyl, tetrahydrofuran base.

Type I compound pharmaceutically acceptable salt refers to: type I compound becomes salt with mineral acid, organic acid.The most preferred: salt Hydrochlorate, hydrobromate, hydriodate, sulfate, disulfate, phosphate, acetate, propionate, butyrate, lactate, first Sulfonate, tosilate, maleate, benzoate, succinate, tartrate, citrate, fumarate, cattle Sulfonate, gluconate, amino acid salts.

The syntheti c route of type I compound is as follows:

R₁, R₂Defined as described above.

In this syntheti c route, the preparation of compound (II) is referred to document Indian Journal of Chemistry, 2006,45B:1699-1703, the general Study personnel of this research field all can conveniently prepare.Specific as follows:

Wherein, the compound (II) of preferential synthesis includes:

Compound (II) and bromoacetyl bromide (III) are dissolved in dichloromethane, chloroform, ethyl acetate, acetone, N, N-diformazan In base Methanamide, oxolane, dioxane, pyridine, benzene, toluene or acetonitrile equal solvent, triethylamine, pyridine, potassium carbonate, carbonic acid Under the catalysis of the acid binding agents such as sodium, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide or potassium tert-butoxide ,-20～30 DEG C anti- Intermediate (IV) should be generated.Intermediate (IV) and intermediate (V) are at dichloromethane, chloroform, ethyl acetate, acetonitrile, N, N- In dimethylformamide, oxolane, dioxane, pyridine, acetone, benzene or toluene, triethylamine, pyridine, potassium carbonate, carbonic acid Under the catalysis of the acid binding agents such as sodium, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide or potassium tert-butoxide ,-20～110 DEG C anti- (I) should be generated.

Reaction prepares various compounds or products therefrom is dissolved in DMF, acetone, methanol, ethanol, DMSO or ether dropping Mineral acid, organic acid can be made into pharmaceutically acceptable salt.

Specifically various compounds are dissolved in the one in ether, DMF, acetone, methanol, ethanol, ethyl acetate or DMSO, Under ice-water bath, dropping ethereal HCI is to pH=2, makes hydrochlorate；Or various compounds are dissolved in ether, DMF, acetone, methanol, One in ethanol, ethyl acetate or DMSO, adds equimolar taurine, and heated and stirred obtains its taurate；Or by various Compound is dissolved in the one in ether, DMF, acetone, methanol, ethanol, ethyl acetate or DMSO, drips concentrated sulphuric acid under ice-water bath To pH=3, make sulfate, etc..

This compounds is effective for treatment human malignancies.Although the compound of the present invention can be without any Preparation is directly administered, but described various compounds use the most in the form of a pharmaceutical preparation, and route of administration can be non-bowel Approach (such as vein, intramuscular delivery) and oral administration.

The pharmaceutical composition preparation of the compounds of this invention is as follows: uses standard and conventional technique, makes the compounds of this invention Acceptable solid or liquid-carrier are combined on galenic pharmacy, and be allowed at random with on galenic pharmacy acceptable adjuvant and Excipient combines and is prepared as microgranule or microsphere.Solid dosage forms includes tablet, discrete particles, capsule, slow releasing tablet, slow-release micro-pill etc. Deng.Solid carrier can be at least one material, and it can serve as diluent, flavouring agent, solubilizing agent, lubricant, suspending agent, glue Mixture, disintegrating agent and coating agent.Inert solid carrier include magnesium phosphate, magnesium stearate, smoothers sugar, lactose, pectin, the third two Alcohol, polyoxyethylene sorbitan monoleate, dextrin, starch, gelatin, cellulose substances such as methylcellulose, microcrystalline Cellulose, low melting point stone Wax, Polyethylene Glycol, mannitol, cocoa butter etc..Liquid dosage form includes solvent, suspension such as injection, powder etc..

The amount of the active ingredient (the compounds of this invention) contained in pharmaceutical composition and unit dosage form can be according to patient The state of an illness, the situation of diagnosis be specifically applied, the amount of compound used or concentration are in a wider scope Regulation, generally, the weight range of reactive compound is 0.5%～90%(weight of compositions).Another is preferably in the range of 0.5%- 70%。

The compound with structure shown in formula I of the present invention or its pharmaceutically acceptable salt, have significantly tumor in vitro Inhibitory action.

External antitumor action:

(1) experimental technique:

Use classical cytotoxic activity vitro detection method mtt assay, people's tumor that detection invention Compounds in vitro is cultivated The cell proliferation toxicity of cell.

(2) experiment material:

Laboratory sample: type I compound is made by oneself offer by inventor.During experiment, sample is trained with DMSO hydrotropy, serum-free DMEM Foster base is diluted to desired concn, and sample segment solution is suspension.

Main agents: MTT, Amresco company subpackage, lot number: 04M0904；DMEM culture medium completely, Gibco company produces Product, lot number: 1290007；Calf serum, Lanzhou people's marine growth, lot number: 20060509；Trypsin, Amresco company subpackage, Lot number: 016B0604；Fluorouracil Injection, 0.25g/10ml (props up), lot number: 0512022, the Tianjin gold credit limited public affairs of aminoacid Department.

Experimental apparatus: superclean bench, Suzhou Decontamination Equipment Plant；CO₂Incubator, Thermo company, model: HERA Cell150；Inverted microscope, Carl Zeiss company, model: Axiovert200；Enzyme-linked immunosorbent assay instrument, TECAN company, Model: Sunrise；Centrifuge, Kerdro company, model: Heraeus.

Cell strain: SPCA1 human lung adenocarcinoma cell line, MCF7 human breast cancer cell, SGC-7901 gastric carcinoma cells, is purchased from Shanghai cell research institute of the Chinese Academy of Sciences.

(3) experimental procedure:

Cell is cultivated: tumor cell inoculation is containing 10% calf serum, 100IU/ml penicillin G sodium salt and 100 μ g/ml sulfur Acid streptomycin DMEM culture fluid in, be placed in 37 DEG C, 100% relative humidity, containing 5%CO₂Incubator in, standby after passing on 3 times.

Mtt assay measures: the cell of trophophase of taking the logarithm, after 0.25% trypsinization (suspension cell need not digest), It is suspended in the DMEM culture fluid containing 10% calf serum, blows and beats into gently with glass dropper and use blood under single cell suspension, microscope Cell count plate numeration living cells.96 well culture plate every hole inoculating cell suspension 90 μ L(cell concentrations are adjusted to 6～10 × 10⁴ Individual/ml), 37 DEG C, 100% relative humidity, containing 5%CO₂, the incubator of 95% air cultivate after 24h, every hole adds 10 μ L medicinal liquids (eventually Concentration is set to: 40 μ g/ml, 20 μ g/ml, 10 μ g/ml, 5 μ g/ml and five concentration of 2.5 μ g/ml).It addition, each concentration sets the moon Property comparison (isoconcentration DMSO) and blank background (being not added with cell), each group is all provided with 6 multiple holes.Cultivate 24h, then every hole the most continuously Adding the MTT solution 10 μ L of 5mg/ml, after continuing to cultivate 4h, (suspension cell needs first to be centrifuged, then inhales carefully to suck supernatant Remove supernatant).Every hole adds 100 μ L DMSO, puts micro oscillator concussion 5min so that crystallization is completely dissolved, and microplate reader 492nm is mono- Wavelength colorimetric, measures OD value.Inhibitory rate of cell growth is calculated as evaluation index using following method.

Suppression ratio (%)=[1-(experimental group OD average-blank group OD average)/(matched group OD average-blank group OD average)] ×100%.According to inhibitory rate of cell growth, calculate IC with rectilinear regression method₅₀Value.

(4) experimental result:

The table 1 IC to the tumor cell of In vitro culture₅₀(μg/ml)

(5) conclusion:

According to above-mentioned in vitro tests result, we can see that the compound with structure shown in formula I is to above-mentioned 3 kinds of human tumors Cell has stronger inhibitory action.

Detailed description of the invention

Below in conjunction with embodiment, the present invention is described further, and embodiment is only explanatory, is in no way intended to it Limit the scope of the present invention by any way.Described compound is carried out through high performance liquid chromatography (HPLC), thin layer chromatography (TLC) Detection.Can use such as infrared spectrum (IR) subsequently, nuclear magnetic resoance spectrum (¹H NMR,¹³C NMR), mass spectrum (MS) etc. more enters one Step confirms its structure.

Reference example 1:

The preparation of ZJ-a

The synthesis of ZJ-a is carried out in microwave synthesizer.By YL-1(16g, 0.1mol) and YL-2a(11ml, 0.1mol) put in mortar and mix, grind to form homogeneous pastel, this pastel is put into microwave synthesizer, react 5min.Instead After should terminating, mixture is poured in frozen water, extracts with chloroform, divide and take organic layer, be dried overnight with anhydrous sodium sulfate.Filter, Decompression distillation obtains brown solid crude product.Gained crude product ethyl alcohol recrystallization, obtains light tan solid 20.4g, fusing point 62.6- 64.3 DEG C, yield is 82.1%.Purity 96.5%(HPLC normalization method), ESI-MS (m/z): 249.1.

With reference to the method for reference example 1, can prepare ZJ-b, ZJ-c, ZJ-d(are shown in Table 2).

Table 2 compound ZJ-b-ZJ-d

Reference example 2:

The preparation of intermediate II-1

By YL-3(0.1mol, 9.6g), NaOH(0.1mol, 4g) and previous step prepare ZJ-1 grind homogeneous, put into micro- Ripple synthesizer, reaction 8min terminates.Reactant mixture is poured in frozen water.Filter, obtain brown solid.Solid ethanol Recrystallization, obtains light brown II-1 product 18.4g, fusing point 73.8-75.1 DEG C, and yield is 77.5%.Purity 97.0%(HPLC normalizing Change method), ESI-MS (m/z): 290.1.

With reference to the method for reference example 1, can prepare II-2, II-3, II-4(are shown in Table 3).

Table 3 compound II-2-II-4:

Embodiment 1:

The preparation of compound IV-1

Equipped with stirring and thermometer three mouthfuls of reaction bulbs in, add 4g(14mmol) intermediate II-1,40ml dichloromethane Alkane, stirring makes it dissolve, and adds triethylamine 2.1g(21mmol), it is cooled to 0 DEG C.By III(3.2g, 15mmol) it is dissolved in 10ml bis- In chloromethanes, it is slowly dropped in reaction system.Keeping temperature stirring 1h, reaction terminates.Reactant liquor is poured in frozen water, divides and take Organic layer, washes three times, is dried.Filtering, distillation under pressure removes solvent, obtains brown solid crude product.Gained crude product silicagel column Chromatography purification.Finally give color solid 4.9g, yield 85%, purity 97.7%(HPLC normalization method), MS (m/z) [M+H]⁺: 411.0。

With reference to the method for embodiment 1, compound IV-2～IV-4(can be synthesized and be shown in Table 4).

Table 4 compound IV-2～IV-4

Embodiment 2:

The preparation of compound I-1

Equipped with stirring and thermometer reaction bulb in, add 1g(2.4mmol) intermediate compound IV-1,40ml acetonitrile, 0.14gNaOH (3.6mmol), acetonitrile (10ml) solution of dropping 0.23g V-1 (2.6mmol), it is stirred at room temperature 8h, reaction Terminate.By reacting liquid filtering, filtrate is poured in water, obtains brown solid crude product.This crude product silica gel column chromatography is purified, obtains Brown product I-1, yield 74%, purity 98.8%(HPLC normalization method), MS (m/z) [M+H]⁺: 416.2.

With reference to the method for embodiment 2, compound I-2～I-24(can be synthesized and be shown in Table 5).

Table 5 compound I-2～I-24

Embodiment 3:

Compounds I-1 hydrochloric acid salt: take compounds I-1 white solid product 0.6g, be dissolved in 8ml dehydrated alcohol.Ice-water bath Being cooled to 5 DEG C, dripping 11.1% ethanol solution hydrochloride to pH is 2, continues at stir about 1h under ice-water bath.Filter, vacuum drying, Obtain white solid powder.

Embodiment 4:

Compounds I-4 becomes taurate: take compounds I-2 white solid product 0.5g(m.p.184.1～185.9 DEG C, HPLC97.6%), 10ml absolute methanol it is dissolved in.Add equimolar taurine after being heated to backflow, continue at stirred at reflux reaction About 1.5h.React complete, stand 24h at room temperature.Separate out Light yellow crystals, filter, vacuum drying.

Embodiment 5:

Compounds I-8 becomes sulfate: takes compounds I-5 white solid product 0.3g, is dissolved in 15ml acetone.Ice-water bath cools down To 0 DEG C, dropping concentrated sulfuric acid solution is 3 to pH, continues at stir about 1h under ice-water bath.Filter, obtain white solid.

In order to be more fully explained the pharmaceutical composition of the pyrimidines containing benzofuran of the present invention, carry below For following example of formulations, described embodiment is merely to illustrate rather than for limiting the scope of the present invention.Described preparation is permissible Use any reactive compound in the compounds of this invention and salt thereof, compound embodiment 2-5 described in is preferably used.

Embodiment 6:

Hard gelatin capsule is prepared by following compositions:

Consumption/capsule

Preparation technology: be pre-dried by supplementary material, crosses 100 mesh sieves standby.After mentioned component being mixed by recipe quantity, fill Enter in hard gelatin capsule.

Embodiment 7:

Tablet is prepared by following compositions:

Consumption/sheet

Preparation technology: be pre-dried by supplementary material, crosses 100 mesh sieves standby.First the adjuvant of recipe quantity is fully mixed.By former Material medicine is added in adjuvant with incremental dilution method, and each added-time fully mixes 2-3 time, it is ensured that medicine fully mixes with adjuvant, crosses 20 mesh Sieve, is dried 2h in 55 DEG C of ventilated drying ovens, and dry granule crosses 16 mesh sieve granulate, measures intermediates content, and mix homogeneously, at tablet machine Upper tabletting.

Embodiment 8:

The preparation of injection:

Preparation method: take in the water for injection that active component joins solubilized solution Polysorbate and propylene glycol, add medicinal Alkali regulation pH value makes it dissolve to 4～8.Add activated carbon, stirring and adsorbing 30min, carbon removal, fine straining, embedding, sterilizing.

Embodiment 9:

The preparation of injection lyophilized powder:

Preparation method: take active component and add water for injection, makes it dissolve with medicinal basic regulation pH value to 4-8.Add Mannitol, carries out autoclaving by the requirement of injection, adds activated carbon, uses filtering with microporous membrane, and filtrate carries out subpackage, adopts With freeze-drying, prepare loose block, sealing, to obtain final product.

Claims (11)

1. there is compound or its pharmaceutically acceptable salt of structure shown in formula I:

Wherein:

R₁For: hydrogen, halogen, methyl, ethyl, methoxyl group；

R₂For: hydrogen, methyl, ethyl, phenyl,Wherein R₃For phenyl, furyl, tetrahydrofuran base.

There is compound or its pharmaceutically acceptable salt of structure shown in formula I the most as claimed in claim 1:

Wherein:

R₁For: hydrogen, fluorine, chlorine, bromine, methyl, ethyl, methoxyl group；

R₂For: hydrogen, methyl, ethyl, phenyl,

Wherein R₃For phenyl, furyl, tetrahydrofuran base.

3. the compound with structure shown in formula I as described in any one of claim 1-2 or its pharmaceutically acceptable salt, is selected from:

4. the compound with structure shown in formula I as described in any one of claim 1-2 or its pharmaceutically acceptable salt, its pharmacy Upper acceptable salt is: compound of formula I becomes salt with mineral acid, organic acid.

Having compound or its pharmaceutically acceptable salt of structure shown in formula I the most as claimed in claim 4, it is pharmaceutically acceptable Salt be: hydrochlorate, hydrobromate, hydriodate, sulfate, disulfate, phosphate, acetate, propionate, butyrate, Lactate, mesylate, tosilate, maleate, benzoate, succinate, tartrate, citrate, richness Horse hydrochlorate, taurate.

6. the preparation method of the compound with structure shown in formula I as described in any one of claim 1-3, it is characterised in that: non- In protonic solvent, compound ii and compound III prepare compound IV, compound IV and chemical combination under acid binding agent effect Thing V prepares type I compound under acid binding agent effect；

R₁, R₂Definition is as described in any one of claim 1-3.

7. preparation method as claimed in claim 6, it is characterised in that: described non-protonic solvent is dichloromethane, three chloromethanes Alkane, ethyl acetate, acetone, N,N-dimethylformamide, oxolane, dioxane, pyridine, benzene, toluene or acetonitrile.

8. preparation method as claimed in claim 6, it is characterised in that: described acid binding agent be triethylamine, pyridine, potassium carbonate, Sodium carbonate, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide or potassium tert-butoxide.

9. a pharmaceutical composition, what it comprised therapeutically effective amount has structure shown in formula I as according to any one of claim 1-2 Compound or its pharmaceutically acceptable salt and one or more pharmaceutical carriers.

10. the compound with structure shown in formula I as according to any one of claim 1-3 or its pharmaceutically acceptable salt with Application in terms of preparing antitumor drug.

11. apply as claimed in claim 10, in the purposes in terms of preparing treatment breast carcinoma, pulmonary carcinoma, gastric cancer medicament.