CN106928246A

CN106928246A - A kind of compound and preparation method thereof and purposes

Info

Publication number: CN106928246A
Application number: CN201710137133.7A
Authority: CN
Inventors: 李卫民; 冯毅凡; 朱贺年; 玉荣
Original assignee: Beijing Yishengtang Medicine Science And Technology Research Co Ltd
Current assignee: Beijing Yishengtang Medicine Science And Technology Research Co Ltd
Priority date: 2017-03-09
Filing date: 2017-03-09
Publication date: 2017-07-07
Anticipated expiration: 2037-03-09
Also published as: CN106928246B

Abstract

The present invention discloses a kind of compound, and the compound is prepared from by ring-closure reaction under certain conditions by berberrubine with melbine.Compound of the present invention is under the guidance of tcm clinical medication experience and on the basis of organic chemistry, purposefully, selectively berberrubine and melbine are combined together by specific synthetic method, gained compound has the effect of Synergy and attenuation, has more preferable effect at the aspect such as anti-inflammatory, hypoglycemic, antitumor, antibacterial, resisting cardiovascular disease.Meanwhile, the invention also discloses the compound preparation method and the compound prepare for treat tumour, hyperglycaemia, high fat of blood, cardiovascular and cerebrovascular disease medicine in purposes.

Description

A kind of compound and preparation method thereof and purposes

Technical field

The present invention relates to a kind of compound and preparation method thereof, especially it is a kind of have anti-inflammatory, antitumor, hypoglycemic, Compounds of effect such as antibacterial and preparation method thereof.

Background technology

Now, with the development of modern pharmacology, molecular biology scheduling theory and related science technology, natural drug is opened Hair approach and means are also being constantly brought forth new ideas.At present, the approach of developing new drug substantially has three, i.e. Folk medicine approach, experience medicine on the way Footpath and molecular pathways.Wherein with the development of molecular biology, science of heredity and the information processing technology etc., molecular pathways gradually into It is the main path of developing new drug.But based on the current national conditions of China, and due to the limitation of technical equipment, current Folk medicine is on the way Footpath, experience medicine approach are the main paths of China's developing new drug, and both approach are based on medicinal plant.

With the medicine that medicinal plant grows up as main body, in history to the preventing and curing diseases of the mankind, rehabilitation and life Educate procreation and serve huge effect.Natural drug available sources are a lot of, mostly from plant；According to estimates, the whole world there are about 40 ~50 ten thousand kinds of plants, but it is only few partly carried out chemical composition and its active testing research, China be also plant resources most One of abundant country, therefore fully excavate and using the new natural drug of the compound exploitation of nature structure diversity, promote Enter China's medical sci-tech to advance at utmost speed, open up new market, promote the development of China's natural drug, meet medication economics to globalize Challenge has great importance.Medicinal plant is the important sources of developing new drug, at present, in the new drug of whole world approval, close to half Number is natural drug, and medicine comes from natural products.Now, also past 60% population also fully relies on autonomic drug to prevent in the world Control disease.

But with the development of disease, the problems such as viral resistance, body drug resistance, simple certain medicinal plant of dependence, Or the new drug that ingredient is researched and developed for precursor in medicinal plant, far from people are met to medicine the need for.For certain Specific disease, people often by two or more, or even more medicines take and can just play certain simultaneously Therapeutic action.

Chinese Drug Rhizomes of Coptis mainly includes the Ranunculaceae coptis, the triangle leaf coptis, cloud dry rhizome even.Berberine scientific name barberry Alkali, is the main alkaloid in Chinese Drug Rhizomes of Coptis.Jamaicin belongs to isoquinoline alkaloid, and Recent study finds, jamaicin and its Derivative is in treatment tumour, diabetes, angiocardiopathy, high fat of blood, inflammation, bacterium and viral infection, cerebral ischemic injury, essence Refreshing disease, Alzheimer disease (senile dementia), osteoporosis etc. are many-sided to have pharmacological action.

Melbine is the classical antidiabetic drug of Western medicine, and Recent study finds that melbine is except with good hypoglycemic Outside effect, also have a good curative effect in anti-inflammatory, antitumor, regulation blood fat, antibiosis, research find the solubility of jamaicin with The increase of pH and increase, during pH13, the Determination of oil-water partition coefficient of jamaicin is maximum, and melbine is strong basicity fatty amine, pH= 12.4, the solubility of jamaicin can be increased, improve bioavilability.Melbine elimination half-life period in vivo is about 1.25- 2.8 hours, its administration number of times is caused to increase, patient compliance is poor, although jamaicin absorption difference in vivo, absorbed into serum Drug distribution is wide, long half time, makes up the shortcoming of melbine half-life short, and jamaicin can improve impaired renal patient's disease Reason change, promotes melbine excretion, reduces lactic acidosis.Based on factors above, the two molecule parent nucleus is spliced one Rise, the bioavilability of jamaicin can be increased, reduce the risk of nephrotic's lactic acidosis, the two is closed using chemistry Into method, it is autotelic to be stitched together, see that can its some pharmacological action strengthen, if 1+1 >=2, Synergy and attenuation can be reached Effect, the two anti-inflammatory, pharmacological action etc. of hypoglycemic drop ester can be protruded, these are all unknown unpredictable, are also existing Having does not have any disclosure or report in technology.

The content of the invention

A kind of new compound is provided it is an object of the invention to overcome above-mentioned the deficiencies in the prior art part, describedization The toxicity of compound than jamaicin small toxicity, and with more preferable anti-inflammatory, antitumor anticancer, hypoglycemic, antibacterial etc. effect. Meanwhile, the present invention also provides the Preparation method and use of the compound.

To achieve the above object, the technical scheme taken of the present invention is：A kind of compound, the structural formula of the compound For：

By berberrubine and melbine by being specifically synthesized, it can be significantly reduced compound of the present invention The toxicity of jamaicin, and 1+1 >=2 can be reached, play a part of Synergy and attenuation, in anti-inflammatory, antitumor anticancer, hypoglycemic There is more preferable effect with the aspect such as antibacterial.

In addition, the present invention discloses a kind of preparation method of compound described above, it is comprised the following steps：

(1) compound shown in formula (1) and bromo agent are carried out into bromo-reaction in solvent orange 2 A, obtains chemical combination shown in formula (2) Thing；

(2) formula (2) compound that will be obtained in step (1) exists in the presence of base catalyst with compound shown in formula (3) There is ring-closure reaction in solvent B, that is, obtain formula of the present invention (4) compound.

The preparation method of compound of the present invention, by specific mode by berberrubine and the parent nucleus ring of melbine It is combined, but this cyclization is not blindness, but under the guidance of tcm clinical medication experience and organise On the basis of, purposefully, selectively the two is combined, it is found that the two can play the effect of Synergy and attenuation after combining Really, and the method for the present invention is simple to operate, compound of the present invention can rapidly and efficiently be obtained.

Used as the preferred embodiment of the preparation method of compound of the present invention, bromo agent is bromine in the step (1) In methyl acetate, bromoacetate, bromoacetic acid butyl ester, methyl chloroacetate, ethyl chloroacetate, butyl chloroacetate, iodoacetic acid methyl esters At least one.As the more preferably implementation method of the preparation method of compound of the present invention, bromo agent in the step (1) It is methyl bromoacetate.When the bromo agent selects methyl bromoacetate, two carbon in the methyl bromoacetate molecule both can be with Overcome intermolecular resistance, it is ensured that the generation of nucleophilic substitution, the stability of synthesized compound maintained again, it is to avoid its It is unstable and decompose in vivo.

Used as the preferred embodiment of the preparation method of compound of the present invention, solvent orange 2 A is second in the step (1) In nitrile, DMF, methyl alcohol, acetone, ethanol, dimethyl sulfoxide (DMSO), tetrahydrofuran, dichloromethane, dimethyl sulfoxide (DMSO), dioxane extremely Few one kind.Used as the more preferably implementation method of the preparation method of compound of the present invention, the solvent orange 2 A in the step (1) is Dichloromethane.When the solvent orange 2 A in the step (1) selects dichloromethane, dichloromethane is molten to the reactant that step (1) is reacted Solution property is preferable, and viscosity is small, low boiling point, and polarity is moderate, on the premise of it both ensure that product yield, post processing is easy to again, returns Receive solvent.

Used as the preferred embodiment of the preparation method of compound of the present invention, base catalyst is in the step (2) At least one in sodium methoxide, caustic alcohol, metallic sodium.As the more preferably embodiment party of the preparation method of compound of the present invention Formula, the base catalyst in the step (2) is sodium methoxide.Present inventor is had found by numerous studies, when the base catalysis During agent selection sodium methoxide, the yield of product is apparently higher than other base catalysts.

Used as the preferred embodiment of the preparation method of compound of the present invention, solvent B is second in the step (2) Nitrile, DMF, ethyl acetate, methyl alcohol, acetone, pyridine, quinoline, dimethyl sulfoxide (DMSO), tetrahydrofuran, dichloromethane, dimethyl sulfoxide (DMSO), At least one in dioxane.As the more preferably implementation method of the preparation method of compound of the present invention, the step (2) the solvent B in is methyl alcohol.The reactant solubility that methyl alcohol is reacted step (2) is preferable, and viscosity is small, low boiling point, and polarity is fitted In, on the premise of it both ensure that product yield, post processing, recycling design are easy to again.

Used as the most preferred embodiment of the preparation method of compound of the present invention, the solvent orange 2 A in the step (1) is Dichloromethane, bromo agent is methyl bromoacetate；Solvent B in the step (2) is methyl alcohol, and base catalyst is sodium methoxide.

As the preferred embodiment of the preparation method of compound of the present invention, solvent orange 2 A and formula in the step (1) (1) mol ratio of compound is 5:1~20:1, bromo agent is 0.8 with the mol ratio of formula (1) compound:1~3:1.

As the preferred embodiment of the preparation method of compound of the present invention, base catalyst and formula in the step (2) (2) mol ratio of compound is 0.04:1~5:1, solvent B are 5 with the mol ratio of formula (2) compound:1~20:1, formula (3) is changed Compound is 0.5 with the mol ratio of formula (2) compound:1~5:1.

As the preferred embodiment of the preparation method of compound of the present invention, bromo-reaction temperature in the step (1) It is 30~80 DEG C to spend, and the reaction time is 3~12 hours.As the more preferably embodiment party of the preparation method of compound of the present invention Formula, bromo-reaction temperature is 30~80 DEG C in the step (1), and the reaction time is 8~10 hours.

As the preferred embodiment of the preparation method of compound of the present invention, ring-closure reaction in the step (2) Temperature is 40~100 DEG C, and the reaction time is 3~20 hours.As compound of the present invention preparation method it is more preferably real Mode is applied, the temperature of ring-closure reaction is 60~70 DEG C in the step (2), the reaction time is 12-20 hours.

Present inventor has found by research, when the bromo-reaction temperature in the step (1) is 40 DEG C, reaction time It is 8h, when the temperature of the ring-closure reaction in step (2) is 65 DEG C, the reaction time is 16 hours, both can guarantee that product yield, has again There is preferable economy.

Used as the preferred embodiment of the preparation method of compound of the present invention, the specific method of the step (1) is： By in the clean there-necked flask of the addition of compound shown in formula (1), solvent orange 2 A is added, bromo agent is slowly added to, when the temperature to 40 DEG C When, back flow reaction, HPLC monitoring reactions stop reaction when raw material fundamental reaction is completely or product is not further added by, and treat that temperature is down to 20-30 DEG C, crystallization, then suction filtration, washing, drying, obtain final product compound shown in formula (2).Preferably, when the crystallization reacts, adopt With adding appropriate 95% ethanol to carry out crystallization, after crystallization after stirring and crystallizing 2h, then carry out suction filtration, washing, dry.

Used as the preferred embodiment of the preparation method of compound of the present invention, the specific method of the step (2) is： By in the clean there-necked flask of the addition of compound shown in formula (3), solvent B and base catalyst are added, stirred 30 minutes, be subsequently adding Compound shown in the formula (2) that step (1) is obtained, reacts 3~20 hours at being 60-70 DEG C in temperature, and reaction carries out cold after terminating But, suction filtration, washing, then dries, then carries out crystallization, has orange-yellow crystal to separate out, and then suction filtration, drying obtains final product institute of the present invention State compound.Preferably, hot methanol extraction is used 5 times after reaction terminates, the temperature of hot methanol is 60 DEG C, after extraction terminates, is sunk Form sediment and add hydrochloric acid-ethanol solution, crystallization, suction filtration is dried, and obtains compound of the invention.

As the preferred embodiment of the preparation method of compound of the present invention, step (1) Chinese style (1) compound It is prepared from using following methods：By compound shown in formula (5) in solvent C at 120~170 DEG C react, make its 9 methyl Slough, obtain compound shown in formula (1)；Wherein described solvent C is DMF, DMAC, DMSO, HMPT, HEPT, NMP, diphenyl ether, pyrrole One kind in pyridine, quinoline, the solvent C is 5 with the mol ratio of compound shown in formula (5):1~20:1；

As the more preferably implementation method of the preparation method of compound of the present invention, shown in the step (1) Chinese style (1) Compound is prepared from using following methods：Compound shown in formula (5) is placed in the there-necked flask of dried and clean, solvent is added C, the heating reflux reaction in electric heating cover is reacted at being 120-170 DEG C in temperature, and course of reaction is monitored with HPLC, raw material base Stop reaction when this reaction is completely or product is not further added by, treat that temperature is down to less than 120 DEG C, pure frozen water is added while stirring, It is placed in and crystallization is stood in refrigerator, temperature is 4 DEG C or so, then suction filtration, washing, is drying to obtain compound shown in formula (1).

Compound shown in step (1) Chinese style (1) of the present invention can directly be purchased from market, it would however also be possible to employ described above Method compound shown in formula (5) is prepared from.

In addition, being prepared for treating tumour, cancer, hyperglycaemia, blood high present invention also offers the above compound Purposes in fat, the medicine of cardiovascular and cerebrovascular disease.

Compound of the present invention be inventor under the guidance of tcm clinical medication experience and organic chemistry, have mesh , selectively berberrubine and melbine are combined together by specific synthetic method and are obtained, compound tool There is the effect of Synergy and attenuation, have more at the aspect such as anti-inflammatory, antitumor, anticancer, hypoglycemic, antibacterial, resisting cardiovascular disease Good effect.

The beneficial effects of the present invention are：Compound of the present invention is by berberrubine and melbine by specific anti- Should synthesize, its toxicity that can significantly reduce jamaicin, and the pharmacological action of 1+1 >=2 can be reached, play Synergy and attenuation Effect；The preparation method of compound of the present invention is simple to operate, can be rapidly and efficiently obtain compound of the present invention, and And gained compound has in the medicine for being used for treating tumour, cancer, hyperglycaemia, high fat of blood, cardiovascular and cerebrovascular disease etc. is prepared Important use, for the medicine for preparing treatment tumour, cancer, hyperglycaemia, high fat of blood, cardiovascular and cerebrovascular disease etc. provides new medicine Selection.

Brief description of the drawings

Fig. 1 is the high resolution mass spectrum figure of compound of the present invention；

Fig. 2 is the infrared spectrogram of compound of the present invention；

Fig. 3 is the ultraviolet spectrogram of compound of the present invention；

Fig. 4 is the proton nmr spectra carbon spectrogram of compound of the present invention；

Fig. 5 is the proton nmr spectra carbon spectrogram of compound of the present invention.

Specific embodiment

For the object, technical solutions and advantages of the present invention are better described, below in conjunction with specific embodiment to the present invention It is described further.

The structural formula difference of the compound of formula used by following examples (1)~(5) is as follows：

Embodiment 1

A kind of embodiment of compound of the present invention, compound described in the present embodiment is prepared from using following methods：

(1) formula (5) compound is placed in the there-necked flask of dried and clean, it is described to DMAC is added in there-necked flask DMAC is 5 with the mol ratio of formula (5) compound:1, the then heating reflux reaction in electric heating cover, reaction temperature is 120-170 DEG C, monitored using HPLC and reacted, when raw material fundamental reaction is complete or product is not further added by, stop reaction；Treat that temperature is down to 120 Below DEG C, pure frozen water being added while stirring, being placed in and crystallization is stood in refrigerator, temperature is 4 DEG C of degree or so, then suction filtration, washing, It is drying to obtain the compound of formula (1)；In this step, the yield of gained formula (1) compound is 75%, and purity is 97%

(2) compound of formula (1) is added in the there-necked flask of dried and clean, adds dichloromethane, the dichloromethane It is 20 with the mol ratio of formula (1) compound:1, methyl bromoacetate is slowly added to, the methyl bromoacetate for being added and formula (1) chemical combination The mol ratio of thing is 0.8:1, temperature rises to 40 DEG C or so, and back flow reaction 8 hours, HPLC monitoring reactions, raw material fundamental reaction is complete Stop reaction when complete or product is not further added by, treat that temperature is down to room temperature, add appropriate 95% ethanol to carry out crystallization, stirred after crystallization After crystallization 2h, then carry out suction filtration, washing, dry, obtain final product the compound of formula (2)；In this step, the receipts of gained formula (2) compound Rate is 85%, and purity is 98%；

(3) by the clean there-necked flask of formula (3) compound addition, methyl alcohol and sodium methoxide-methanol solution are added, is added The mol ratio of methyl alcohol and formula (2) compound be 20:1, the sodium methoxide for being added is 0.04 with the mol ratio of formula (2) compound: 1, stir 30 minutes, the compound of the formula (2) that step (1) is obtained is slowly added to, the formula for being added (3) compound is changed with formula (2) The mol ratio of compound is 0.5:1, reacted 16 hours at being 65 DEG C in temperature, cooling, suction filtration, drying after terminating are reacted, obtain formula (4) compound, obtains final product compound of the present invention；The compound of gained formula (4) in this step, yield is 33.5%, purity It is 49%, is more than 95% by purity after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol, obtains final product of the inventionization Compound.

Embodiment 2

(1) formula (5) compound is placed in the there-necked flask of dried and clean, it is described to DMSO is added in there-necked flask DMSO is 10 with the mol ratio of formula (5) compound:1, then in electricity set in heating reflux reaction, reaction temperature be 120-170 DEG C, Monitored using HPLC and reacted, when raw material fundamental reaction is complete or product is not further added by, stop reaction；Treat that temperature is down to 120 DEG C Hereinafter, pure frozen water being added while stirring, being placed in and crystallization is stood in refrigerator, temperature is 4 DEG C or so, then suction filtration, washing, dry Obtain final product the compound of formula (1)；In this step, the yield of gained formula (1) compound is 72%, and purity is 96%；

(2) compound of formula (1) is added in the there-necked flask of dried and clean, adds dichloromethane, heating, the dichloro Methane is 20 with the mol ratio of formula (1) compound:1, methyl chloroacetate is slowly added to, the methyl chloroacetate for being added and formula (1) The mol ratio of compound is 1:1, temperature rises to 40 DEG C or so, back flow reaction 8 hours, HPLC monitoring reactions, raw material fundamental reaction Stop reaction when complete or product is not further added by, treat temperature room temperature, add appropriate 95% ethanol to carry out crystallization, stirred after crystallization and analysed After brilliant 2h, then carry out suction filtration, washing, dry, obtain final product the compound of formula (2)；In this step, the yield of gained formula (2) compound It is 80%, purity is 98%；

(3) by the clean there-necked flask of the compound addition of formula (3), methyl alcohol and caustic alcohol-ethanol solution are added, it is added The ethanol for entering is 20 with the mol ratio of formula (2) compound:1, the caustic alcohol for being added is with the mol ratio of formula (2) compound 0.04:1, stir 30 minutes, it is slowly added to the compound of the formula (2) that step (1) is obtained, the formula for being added (3) compound and formula (2) mol ratio of compound is 3:1, reacted 16 hours at being 65 DEG C in temperature, cooling, suction filtration, drying after terminating are reacted, obtain The compound of formula (4), obtains final product compound of the present invention；By after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol, Obtain final product the compound of the present invention that purity is more than 95%；The compound of gained formula (4) in this step, yield is 30%, pure It is 49% to spend, by purity after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol be more than 95%.

Embodiment 3

(1) formula (5) compound is placed in the there-necked flask of dried and clean, to adding DMF, the DMF in there-necked flask It is 15 with the mol ratio of formula (5) compound:1, then in electricity set in heating reflux reaction, reaction temperature be 120-170 DEG C, use HPLC monitoring reactions, when raw material fundamental reaction is complete or product is not further added by, stop reaction；Treat that temperature is down to less than 120 DEG C, Pure frozen water being added while stirring, being placed in and crystallization is stood in refrigerator, temperature is 4 DEG C or so, then suction filtration, washing, are drying to obtain The compound of formula (1)；In this step, the yield of gained formula (1) compound is 71%, and purity is 97%；

(2) compound of formula (1) is added in the there-necked flask of dried and clean, adds dichloromethane, heating, the dichloro Methane is 20 with the mol ratio of formula (1) compound:1, methyl chloroacetate is slowly added to, the methyl chloroacetate for being added and formula (1) The mol ratio of compound is 3:1, temperature rises to 40 DEG C or so, back flow reaction 8 hours, HPLC monitoring reactions, raw material fundamental reaction Stop reaction when complete or product is not further added by, treat that temperature is down to room temperature, add appropriate 95% ethanol to carry out crystallization, stirred after crystallization After mixing crystallization 2h, then carry out suction filtration, washing, dry, obtain final product the compound of formula (2)；In this step, gained formula (2) compound Yield is 80%, and purity is 97%；

(3) metallic sodium is wrapped with tinfoil, hole is pricked with pin, in input methanol solution, treat that metallic sodium all dissolves, by tinfoil Take out, solution for standby, during the compound of formula (3) added into clean there-necked flask, with the reacted methanol solution of metallic sodium, institute The methanol solution of addition is 20 with the mol ratio of formula (2) compound:1, the metallic sodium and the mol ratio of formula (2) compound for being added It is 0.04:1, stir 30 minutes, be slowly added to the compound of the formula (2) that step (1) is obtained, the formula for being added (3) compound with The mol ratio of formula (2) compound is 5:1, reacted 16 hours at being 65 DEG C in temperature, cooling, suction filtration, drying after terminating are reacted, obtain To the compound of formula (4), the compounds of this invention is obtained final product；By after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol, obtaining To the compound of the present invention that purity is 97%；The compound of gained formula (4) in this step, yield is 28%, and purity is 49%, it is 97% by purity after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol, obtain final product compound of the invention.

Embodiment 4

(1) formula (5) compound is placed in the there-necked flask of dried and clean, it is described to HMPT is added in there-necked flask HMPT is 20 with the mol ratio of formula (5) compound:1, then in electric heating cover set in heating reflux reaction, reaction temperature is 120- 170 DEG C, monitored using HPLC and reacted, when raw material fundamental reaction is complete or product is not further added by, stop reaction；Treat that temperature is down to Less than 120 DEG C, pure frozen water is added while stirring, is placed in and crystallization is stood in refrigerator, temperature is zero degree or so, then suction filtration, wash Wash, be drying to obtain the compound of formula (1)；In this step, the yield of gained formula (1) compound is 66%, and purity is 98%；

(2) compound of formula (1) is added in the there-necked flask of dried and clean, adds acetonitrile, heating, the acetonitrile and formula (1) mol ratio of compound is 5:1, bromoacetate is slowly added to, the bromoacetate for being added rubs with formula (1) compound You are than being 0.8:1, temperature rises to 70 DEG C or so, and back flow reaction 6 hours, HPLC monitorings are reacted, and raw material fundamental reaction is complete or produces Stop reaction when thing is not further added by, treat that temperature is down to room temperature, add appropriate 95% ethanol to carry out crystallization, stirring and crystallizing 2h after crystallization Afterwards, then suction filtration, washing are carried out, is dried, the yield of gained formula (2) compound is 78%, and purity is 92%；

(3) compound of formula (3) is added into clean there-necked flask, add methanol/ethanol mixed solution and sodium methoxide- Methanol solution, the methanol/ethanol mixed solution for being added is 20 with the mol ratio of formula (2) compound:1, the sodium methoxide that is added with The mol ratio of formula (2) compound is 0.05:1, stir 30 minutes, it is slowly added to the compound of the formula (2) that step (1) is obtained, institute Formula (3) compound of addition is 3 with the mol ratio of formula (2) compound:1, to be reacted 16 hours at being 65 DEG C in temperature, reaction terminates Cooling, suction filtration, drying afterwards, obtains the compound of formula (4), obtains final product the compounds of this invention；By hot methanol washing, hydrochloric acid-ethanol After etc. method polishing purification, the compound of the present invention that purity is 97% is obtained；The compound of gained formula (4) in this step, Yield is 30%, and purity is 49%, by purity after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol be 97%.

Embodiment 5

A kind of embodiment of compound of the present invention, compound described in the present embodiment is prepared from using following methods： (1) formula (5) compound is placed in the there-necked flask of dried and clean, to adding pyridine in there-necked flask, the pyridine and formula (5) The mol ratio of compound is 8:1, the then heating reflux reaction in electric heating cover, reaction temperature is 120-170 DEG C, using HPLC Monitoring reaction, when raw material fundamental reaction is complete or product is not further added by, stops reaction；Treat that temperature is down to less than 120 DEG C, while stirring Mix side and add pure frozen water, be placed in and crystallization is stood in refrigerator, temperature is 4 DEG C or so, then suction filtration, washing, are drying to obtain formula (1) Compound；In this step, the yield of gained formula (1) compound is 72%, and purity is 95%；

(2) compound of formula (1) is added in the there-necked flask of dried and clean, adds tetrahydrofuran, heating, the tetrahydrochysene Furans is 20 with the mol ratio of formula (1) compound:1, bromoacetate is slowly added to, the bromoacetate for being added and formula (1) The mol ratio of compound is 0.8:1, temperature rises to 70 DEG C or so, and back flow reaction 6 hours, HPLC monitoring reactions, raw material is substantially anti- Answer complete or product to stop reaction when not being further added by, treat that temperature is down to room temperature, add appropriate 95% ethanol to carry out crystallization, after crystallization After stirring and crystallizing 2h, then carry out suction filtration, washing, dry, the yield of gained formula (2) compound is 75%, and purity is 92%；

(3) by the clean there-necked flask of the compound addition of formula (3), tetrahydrofuran and sodium methoxide-methanol solution are added, The tetrahydrofuran for being added is 20 with the mol ratio of formula (2) compound:1, the sodium methoxide that is added and formula (2) compound mole Than being 0.1:1, stir 30 minutes, be slowly added to the compound of the formula (2) that step (1) is obtained, the formula for being added (3) compound with The mol ratio of formula (2) compound is 4:1, reacted 16 hours at being 65 DEG C in temperature, cooling, suction filtration, drying after terminating are reacted, obtain To the compound of formula (4), the compounds of this invention is obtained final product；The compound of gained formula (4) in this step, yield is 31%, and purity is 49%, by purity after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol be 95%.

Embodiment 6

(1) formula (5) compound is placed in the there-necked flask of dried and clean, it is described to diphenyl ether is added in there-necked flask Diphenyl ether is 12 with the mol ratio of formula (5) compound:1, the then heating reflux reaction in electric heating cover, reaction temperature is 120- 170 DEG C, monitored using HPLC and reacted, when raw material fundamental reaction is complete or product is not further added by, stop reaction；Treat that temperature is down to Less than 120 DEG C, pure frozen water is added while stirring, is placed in and crystallization is stood in refrigerator, temperature be 4 DEG C or so, then suction filtration, wash Wash, be drying to obtain the compound of formula (1)；In this step, the yield of gained formula (1) compound is 68%, and purity is 96%；

(2) during the compound of formula (1) to add the there-necked flask of dried and clean, the mixed of dichloromethane and dioxane is added In the mixture of compound, the dichloromethane and dioxane, the mass ratio of the dichloromethane and dioxane is 2:1, plus The mixture of heat, the dichloromethane and dioxane is 10 with the mol ratio of formula (1) compound:1, it is slowly added to bromoacetic acid fourth Ester, the bromoacetic acid butyl ester for being added is 2 with the mol ratio of formula (1) compound:1, temperature rises to 100 DEG C or so, and back flow reaction 5 is small When, HPLC monitoring reactions stop reaction when raw material fundamental reaction is completely or product is not further added by, and treat that temperature is down to room temperature, add Appropriate 95% ethanol carries out crystallization, after crystallization after stirring and crystallizing 2h, then carries out suction filtration, washing, dries, and obtains final product the chemical combination of formula (2) Thing；In this step, the yield of gained formula (2) compound is 70%, and purity is 95%；

(3) by the clean there-necked flask of the compound addition of formula (3), tetrahydrofuran and sodium methoxide-methanol solution are added, The tetrahydrofuran for being added is 5 with the mol ratio of formula (2) compound:1, the sodium methoxide that is added and formula (2) compound mole Than being 0.1:1, stir 30 minutes, be slowly added to the compound of the formula (2) that step (1) is obtained, the formula for being added (3) compound with The mol ratio of formula (2) compound is 5:1, reacted 16 hours at being 65 DEG C in temperature, cooling, suction filtration, drying after terminating are reacted, obtain To the compound of formula (4)；The compound of gained formula (4) in this step, yield is 25%, and purity is 49%, is washed by hot methanol Wash, purity is 98% after the method polishing purification such as hydrochloric acid-ethanol..

Embodiment 7

(1) compound of formula (1) is directly bought from market, its purity is 99%；

(2) compound of formula (1) is added in the there-necked flask of dried and clean, adds the mixture of acetonitrile and methyl alcohol, institute State in the mixture of acetonitrile and methyl alcohol, the mass ratio of the acetonitrile and methyl alcohol is 1:4, heating, the mixing of the acetonitrile and methyl alcohol Thing is 8 with the mol ratio of formula (1) compound:1, bromoacetate is slowly added to, the bromoacetate for being added and formula (1) chemical combination The mol ratio of thing is 2:1, back flow reaction 12 hours at being 67 DEG C in temperature, HPLC monitoring reactions, raw material fundamental reaction completely or Stop reaction when product is not further added by, add appropriate 95% ethanol to carry out crystallization, after crystallization after stirring and crystallizing 2h, then taken out Filter, wash, dry, obtain final product the compound of formula (2)；In this step, the yield of gained formula (2) compound is 80%, and purity is 95%；

(3) by the clean there-necked flask of the compound addition of formula (3), the mixture and N, N- bis- of acetonitrile and DMF are added Wopropyl ethyl amine, in the acetonitrile and the mixture of DMF that are added, acetonitrile is 1 with the mass ratio of DMF:1, the acetonitrile and DMF's Mixture is 6 with the mol ratio of formula (2) compound:1；The N, N- diisopropylethylamine is with the mol ratio of formula (2) compound 2:1, sodium methoxide is added, stir 30 minutes, the compound of the formula (2) that step (1) is obtained is slowly added to, the formula for being added (3) is changed Compound is 0.5 with the mol ratio of formula (2) compound:1, to be reacted 16 hours at being 65 DEG C in temperature, reaction is cooled down after terminating, taken out Filter, dry, obtain the compound of formula (4), obtain final product the compounds of this invention；The compound of gained formula (4) in this step, yield is 29%, purity is 49%, by purity after the method polishing purifications such as hot methanol washing, hydrochloric acid-ethanol be 95%.

The compound collection of illustrative plates of the present invention of embodiment 8

Be can be seen that by accompanying drawing 2~5, compound of the invention, m.p.210.5~214.9 DEG C, ESI-MS m/z: 471.1666[M]⁺；

IR(KBr)ν_max:1674.87,1600.63,1533.38,1491.67,1405.85,1237.11,1038.48, 935.306,772.351(cm^-1)；

¹HNMR(500MHz,DCOOD),δ(ppm)7.173(s,2H,O-CH₃-O),6.952(s,1H,C₁₂-H),6.745 (s,1H,C8-H,),6.417(d,2H,C₁-H,C₄-H),5.843(s,1H,C₁₃-H),5.486(s,2H,CH₂-O),5.026(d, 2H,C₆-H,),3.107(s,3H,OCH₃),2.219(s,3H,N-CH₃),2.174(d,5H,C₅-H,N-CH₃)；

¹³CNMR(MHz,DCOOD),δ(ppm):166.56(OCH₂O),161.46(C₂),159.79(C_5a),156.49(N- C-N),156.03(N-C-N)151.41C_1a), 148.97 (N-C=N), 147.80 (C₁),143.05(C_7a),142.21(C₁₀), 132.76(C₁₂),131.08(C_9a),130.94(C_12a),123.97(C₉),121.23(C₂),120.55(C₁₁),108.95 (C₁₃),106.45(C₈),103.02(O-CH₂),102.50(C₄),58.36(OCH₃),53.72(C₆),38.16(N-CH₃), 37.94(N-CH₃),27.15(C₅)。

The antiphlogistic effects experiment of the compound of the present invention of embodiment 9

1st, experiment material, reagent and instrument

Experiment material：RAW264.7 macrophages (are purchased from Chinese Academy of Sciences's cell bank), hyclone (FBS, Gibco)； High glucose medium (DMEM, Gibco), 0.25% trypsase (Gibco)；Pen .- Strep is dual anti-(Gibco)；Phosphate Buffer solution (PBS, Hyclone)；MTT powder (Sigma)；DMSO(Sigma)；Brufen (Sigma)；LPS (Sigma), amino Guanidine (Chinese food drug assay research institute)

Key instrument：HHS types electric-heated thermostatic water bath (Shanghai Boxun Industrial Co., Ltd.)；KDC-220HR is freezed at a high speed Centrifuge (Keda Innovation Co., Ltd), TDL80-2B low speed centrifuges (Anting Scientific Instrument Factory, Shanghai)；BP210D points Analysis balance (Sartorius)；HF90CO₂Incubator (Shanghai Lishen Scientific Equipment Co., Ltd.)；Microscope；SW-CJ-1FD surpasses Net platform (Su Jing is safe and sound)；WH-2 micro-whirlpools mixed instrument (Shanghai Hu Xi analytical instrument factory)；BX51 fluorescence microscopes (OLYMOUS)。

2nd, the preparation of solution

The configuration of cell culture medium：DMEM：FBS：Dual anti-=98：10：1, the configuration of cells frozen storing liquid：FBS：DMSO=9： The compound method of 1, MTT solution:MTT 500mg are weighed, is dissolved in the PBS of 100ml, the MTT concentration of configuration is 5mg/ml, is used 0.22 μm of membrane filtration is put -20 DEG C and is kept in dark place to remove the bacterium in solution.

3rd, cell culture

Cell recovery：1. the cell line for freezing is taken out from liquid nitrogen container or -86 DEG C of ultra low temperature freezers, 37 DEG C is inserted rapidly In thermostat water bath, constantly shaking cryopreservation tube completes operation to melting completely in 1-2min；2. the sterilization of 75% alcohol wipe is outer Cell suspension is gone to sterile centrifugation tube by pipe, rapid transposition super-clean bench, adds 10 times of DMEM culture mediums containing 10%FBS, is blown Beat uniform；3. 1000rpm/min, is centrifuged 5-8min, and abandoning supernatant adds appropriate configured culture medium, is inoculated in 60mm In culture dish, 37 DEG C, 5%CO are placed in₂And cultivated in the incubator of saturated humidity；4. culture medium is changed within every two days.

Passage：1. old culture medium is discarded；2. PBS 2 times, suction is abandoned, and adds appropriate Trypsin Induced 1min, examines under a microscope discovery cytoplasm retraction, space between cells increase, and after cell appearance is rounded, trypsase is abandoned in suction, is stood Add and matched somebody with somebody culture medium；3. ware bottom being blown and beaten repeatedly, attached cell is blown off completely, piping and druming is uniform, and gentle manipulation is avoided the occurrence of Foam；4. with 1:3 ratio is passed on.

Cell freezes：1. -3. same passage of step；4. in the cell suspension for blowing off being gone into sterile centrifugation tube, 1000rpm/min, is centrifuged 5-8min, abandoning supernatant, adds and appropriate has matched somebody with somebody frozen stock solution；5. each cryopreservation tube liquid 1-1.5ml, Final concentration of cells is (5-10) × 10⁶/ml；6. cryopreservation tube is sealed with sealed membrane, carries out mark, is frozen according to following procedure：4 DEG C, 15-20min → -20 DEG C, 30-40min, cell suspension is in after freezing, you can is put into -86 DEG C of ultra low temperature freezers and preserves, for a long time Preservation can be put into liquid nitrogen container.

(1) mtt assay determines the compounds of this invention and RAW264.7 cell-proliferation activities is influenceed

1st, experimental principle

The entitled 3- (4,5- dimethylthiazole -2) of MTT chemistry -2,5- diphenyltetrazolium bromide bromides, trade name：Tetrazolium bromide.It is A kind of dyestuff of yellow color.It is a kind of method for detecting cell survival and growth.Its Cleaning Principle is in living cells mitochondria Succinate dehydrogenase can make exogenous MTT be reduced to the bluish violet Jie Jing formazans of water-insoluble and be deposited in cell, and dead thin Born of the same parents are without this function.Dimethyl sulfoxide (DMSO) (DMSO) can dissolve the first a ceremonial jade-ladle, used in libation in cell, be surveyed at 570nm wavelength with enzyme-linked immunosorbent assay instrument Fixed its absorbance value, can indirectly reflect living cells quantity.In the range of certain cell number, MTT crystallizes the amount and cell number to be formed It is directly proportional.The method is widely used in cell toxicity test, and its feature is that sensitivity is high, economy.

2nd, experimental technique

RAW264.7 cell suspensions are inoculated in 96 orifice plates, overnight incubation with 5000/100 μ L.Compound concentration is used Doubling dilution, 6 concentration gradients, 3 multiple holes of each concentration, the μ L of medicine 100 cultures 48 diluted per hole addition culture medium After hour, 10%MTT is added in every hole, 4h is cultivated at continuing at 37 DEG C, abandoning supernatant adds DMSO150 μ L, uses ELIASA The absorption value at 570nm is read, is 100% with blank, calculate influence of each dosage of compound to cell viability, use spss Software calculates the IC50 values of each compound.

	Brufen	The compounds of this invention	Berberine	Melbine
					IC₅₀(μmol/L)	55.67±4.13	82.36±2.45	65.13±5.62	60.25±8.33

Experimental result shows IC of the compounds of this invention than each monomeric compound and positive control drug brufen before synthesis₅₀Value Greatly, show that its cell toxicant toxicity is reduced.

(2) the compounds of this invention is to LPS induction RAW264.7NO release inhibitory action

Griess method principles：The NO of Hemapoiesis is oxidized easily as NO₂-、NO₃-, first by NO₃- reduction turns into NO₂-, using diazo-reaction and even nitridation reaction generation color compound, there is absorption maximum at 540nm wavelength.Extinction Value and NO₂- concentration is proportionate, so as to obtain NO burst sizes indirectly.

Investigate the change of cell supernatant nitrite after compound is intervened：RAW264.7 cell suspensions are with every hole 1×10⁵Individual/100 μ L are inoculated in 96 orifice plates.Blank control group (only RAW264.7 cells) is set, and (RAW264.7 is thin for model group Born of the same parents and LPS), and drug-treated group (RAW264.7 cells, LPS, medicine), first processed using different pharmaceutical concentration with compound After (100 μM -3 μM) are processed 2 hours, then add LPS, stimulate after maintaining 24h, draw the μ L of nutrient solution 100 in 96 orifice plates, addition etc. The Griess reaction reagents of volume, room temperature reaction 10min reads absorbance value at ELIASA 540nm.Calculate each dosage of compound In the different disposal time to cell supernatant nitrite inhibiting rate.Inhibiting rate (%)=(model group absorbance-each dosage Drug-treated group absorbance)/(model group absorbance-blank control group absorbance) × 100%.And calculated with spss softwares The IC that each compound suppresses to NO₅₀Value.Note:All of compound with DMSO dissolve, and with culture medium be diluted to required for Concentration, the ultimate density of DMSO is 0.1%, and 0.1% DMSO is added in blank control group and model group, is as a result shown to thin Born of the same parents do not have toxic action.Aminoguanidine is used as positive control drug, parallel laboratory test three times.Experimental result GraphPad Prism softwares Analysis.P<0.05, P<0.01 expression result has significant difference.IC₅₀Calculate with Probit programs in SPSS13.0 softwares.

	Brufen	The compounds of this invention	Berberine	Melbine
					IC₅₀(μmol/L)	27.25±4.57	36.42±1.13	41.93±5.12	58.64±2.78

Experimental result display the compounds of this invention IC₅₀Value suppresses NO release actions and increases less than monomeric compound before synthesis By force, but still it is weaker than positive control drug.

The antibacterial effect experiment of the compound of the present invention of embodiment 10

(1) experiment material, laboratory apparatus, reagent

1st, experiment material

Bacterial strain (is clinically separated resistance candida albicanses 103,100,953 and J28 to be given by Changhai hospital Culture Collection Send), all experiments bacterial strain draws plate activation in husky fort glucose agar medium (SDA), after being cultivated 2 weeks in 30 DEG C, respectively Picking monoclonal draws plate activation again, takes second gained monoclonal and puts SDA inclined-planes, cultivates in aforementioned manners after 4 DEG C of preservations With standby.

(1) nutrient solution

The preparation of the liquid mediums of RPMI 1640：RPMI 1640 (Gibco BRL) 10g, NaHCO₃2.0g, morpholine Propane sulfonic acid (MOPS) (Sigma) 34.5g (0.165M), plus tri-distilled water 900ml dissolvings, 1N NaOH adjust pH to 7.0 (25 DEG C), three Steam water and be settled to 000ml, 0.22 μm of filtering with microporous membrane is degerming, packing is saved backup after 4 DEG C.

(2) husky fort agar glucose solid medium (SDA)

Peptone 10g, glucose 40g, agar 18g, plus tri-distilled water 900ml dissolve, and adjust PH to 7.0, fixed with tri-distilled water Hold to 1000ml, autoclaving (121 DEG C, 15min) is saved backup after 4 DEG C.

(3) YEPD nutrient solutions

Yeast extract 10g, peptone 20g, glucose 20g, plus tri-distilled water 900ml dissolve, and tri-distilled water is settled to 1000ml, Autoclaving (121 DEG C, 15min) is saved backup after 4 DEG C.

2nd, experiment reagent

Fluconazole (Fluconazole, FCZ) parenteral solution is provided by Dalian pharmaceutcal corporation, Ltd of Pfizer, Berberine hydrochloride (Berberine Chloride, BBR) is provided by Shanghai Changhai Hospital, dimethyl sulfoxide (DMSO) (Dimethyl Sulphoxide, DMSO) Solution on Chemical Reagents in Shanghai company of Chinese Medicine group produces.

3rd, laboratory apparatus

Multiskan MK3 type enzyme marks detector (Finland Labsystems products), water isolation type electro-heating standing-temperature cultivator (on Sea leap medical apparatus and instruments factory), MJX types intelligence mold incubator (Ningbo south of the River instrument plant) THZ-82A Desk type constant-temperatureoscillator oscillators (Shanghai leap medical apparatus and instruments factory)；SW-CT-IF types superpurgative working table (SuZhou Antai Air Tech Co., Ltd.)；It is inverted aobvious Micro mirror (amersham Pharmacia products)；Micro sample adding appliance (Finland Finnpette products)；96 porocyte culture plates are (red Wheat Nunclon Products)

(2) experimental technique

1st, the preparation of fungi suspension

It is a small amount of from the various candida albicans of picking on 4 DEG C of SDA culture mediums of preservation with inoculation circle before experiment, it is seeded to 1ml YEPD nutrient solutions, in 30 DEG C, 200rpm shaken cultivations, activation 16h makes fungi be in later stage exponential phase of growth.Take the bacterium solution extremely In 1ml YEPD nutrient solutions, counted with blood cell counting plate, with RPMI RPMI-1640s after activation 16h again in aforementioned manners Adjust bacterial concentration to 1 × 10³-5×10³CFU/ml。

2nd, the preparation of drug sensitive reaction plate

Aseptic 96 orifice plate is taken, adds the μ l of 1640 fluid nutrient mediums of RPMI 100 to make blank in No. 1 hole of every row；3-12 holes Respectively plus Fresh the μ l of bacterium solution 100；No. 2 holes add the μ l of the bacterium solution 160 and μ l of test-compound solution 40 respectively；No. 12 holes not pastille Thing, only adds the μ l of bacterium solution 100 to make Growth positive control.2-11 holes carry out doubling dilution, distinguish the final drug concentration in each hole It is 64,32,16,8,4,2,1,0.5,0.25 and 0.125 μ g/ml, DMSO contents are below 1% in each hole.Susceptibility is prepared every time A Quality Control bacterium drug sensitive plate is prepared while plate, each drug sensitive plate is in 30 DEG C of insulating box cultures.

3rd, the judgement of minimum inhibitory concentration (MIC)

In 30 DEG C of insulating boxs, each hole OD values are surveyed in 620nm with enzyme micro-plate reader after candida albicans culture 24h.Positive control 0.2 or so, with positive control boring ratio, the medicine declined with OD values in more than 80% least concentration hole is dense for the OD values control in hole It is MIC to spend₈₀(drug concentration when fungi growth 80% is suppressed).As the MIC of medicine₈₀When value exceedes measure concentration range, press Following methods are counted：MIC₈₀When value is higher than 64 μ g/ml of maximum concentration, be calculated as ">64μg/ml”；MIC₈₀It is least concentration to be worth Or when below least concentration, do not make difference, it is calculated as "≤0.125 μ g/ml ".The equal operation repetitive of above-mentioned experiment 2 to 3 times, when MIC₈₀Value can accurately be repeated or just received when only differing from a concentration, and using higher concentration as MIC₈₀Value；Work as MIC₈₀Value difference When more than two concentration, then need to test again, untill meeting the requirements.

	Penicillin	The compounds of this invention	Berberine	Melbine
					MIC₈₀(μg/ml)	1.06±0.61	5.28±0.83	3.82±0.32	10.54±1.38

From MIC₈₀Value is as can be seen that the compounds of this invention is not as good as alone berberine but remote good to the antibacterial effect of the bacterium In alone melbine curcumin.

The antitumor anticancer effect test of the compound of the present invention of embodiment 11

By taking hepatoma cell strain HepG 2 and leukemia cell line K 562 as an example, by inquiring into the compounds of this invention to liver cancer The antitumor anticancer of the compound pair is studied in the inhibitory action of cell line HepG 2 and leukemia cell line K 562 propagation Effect.

This experiment is using Sulforhodamine B (SRB) dyeing, trypan blue row's dye method, comet electrophoresis technology and AO/EB Toxicity, suppression cancer cell increasing of double dye method detection the compounds of this invention to hepatoma H22 cells and malignant myeloid cell lines K562 The apoptotic effect of the effect of growing, the damage to K562 cell DNAs and induction K562 cells.

(1) material, instrument and reagent

1st, material and reagent

(DMSO prepares stoste to the compounds of this invention, dilutes before use, it is ensured that DMSO concentration at least dilutes in reaction system It is 1 × 10^-2), human hepatoma cell strain He pG2 and human leukemia cell line K562 are quoted from Lanzhou University's Life Science College, culture Base RPM I1640 (U.S. Gibcobr l), yellow acyl rhodamine B (Sulforhodam ineB, SRB, U.S. Sigma), trypan blue (Sigma companies), calf serum (Hangzhou Chinese holly), it is pure that other chemical reagent are analysis.

2nd, laboratory apparatus

U.S. Precision Scientific CO₂Incubator, superclean bench, inverted microscope, light microscope, ELIASA (BIO-RADModel 550 and BIO-RAD Model 680), fluorescence microscope, Horizontal electrophoresis tank, high-speed low temperature from Scheming, Costar Tissue Culture Plates.

(2) experimental technique

Cell culture：During HepG 2 and K562 cells are inoculated in containing 10% calf serum RPM I1640 culture mediums respectively, put 37 DEG C, 5%CO₂Cultivated in incubator, growth period cell of taking the logarithm is used to test.

1st, toxic action of the srb assay detection the compounds of this invention to HepG2 cells

Principle：SulforhodamineB (SRB) is a kind of protein combination dye, pink, water-soluble.SRB can be with Combined with the basic amino acid in large biological molecule, it is in good linear relationship in the OD readings and cell number of 511nm.Therefore can Quantifying as cell number.

Method：The HepG2 cells of exponential phase are with 10 × 10⁴ml^-1Concentration be inoculated in 96 well culture plates, cultivate After 24h, to the μ l of the compounds of this invention fresh medium 100 that different diluted concentrations are separately added into respective aperture, continue to cultivate 24, After 48h, nutrient solution is discarded, add the 10% μ l of trichloroacetic acid (TCA) 100, after standing 5min, by 4 DEG C of fixed 1h of orifice plate dislocation, Fixer is outwelled, is washed with deionized water 5 times, dried.The SRB dyeing 10min of 100 μ l0.4% are added per hole afterwards, with 1% Acetic acid is washed 5 times, is dried naturally.Then 150 μ l 10mmolL are added per hole^-1Non-buffered Tris alkali lye (unbuffered Tris-base solution, pH10.5) SRB combined with protein is dissolved, shaking 5min with oscillator makes it fully dissolve Afterwards, the OD values in every hole are determined at 515nm with ELIASA.Seek its cell proliferation rate (%)=(T-T₀)/(C-T₀)×100.Wherein C represents the cell OD values of control group；T represents the cell OD values of dosing group；T₀Represent cell OD during control flat board measure dosing Value.The cell that will be inoculated with flat board at once before dosing is fixed with TCA.If the final OD values of dosing group are more than T₀；Illustrate thin Born of the same parents still grow after dosing.If the final OD values of dosing group are less than T₀, cell is killed after illustrating dosing.Using recurrence side Journey obtains drug concentration (the 50%inhibition concentration, IC of 50% increment inhibiting rate₅₀).From experimental result See, the final cell number of dosing group is less than T₀, then illustrating that medicine cell growth has inhibitory action, cell is killed after dosing Extremely.With the IC after the compounds of this invention group and control group treatment HepG 2 cell 24h₅₀Carry out each compound of comparing as a example by value thin to this The toxic action of born of the same parents, IC₅₀Value is as shown in the table：

	Cis-platinum	The compounds of this invention	Berberine	Melbine
					IC₅₀(μmol/L)	9.54±0.62	20.53±1.85	22.35±1.42	55.38±3.85

Experimental result display the compounds of this invention IC₅₀Value is less than monomeric compound before synthesis, i.e., to the Carbazole alkaloids of HepG 2 Effect enhancing, but still it is weaker than positive control drug.

2nd, toxic action of trypan blue row's dye method detection the compounds of this invention to K562 cells

Principle：Living cells has the ability for repelling trypan blue, and due to the destruction of film completeness after cell death, cell is quilt Coloring.By in the cell suspension instillation white blood cell count(WBC) plate cell containing trypan blue, observation under an optical microscope counts living thin Born of the same parents (are not dyed to the cell of blueness).

Experimental technique：Concentration is 10 × 10⁴·ml^-1The K562 cell suspensions in exponential phase, with 0.5ml Hole^-1It is inoculated in 24 well culture plates.It is placed in 37 DEG C, %CO₂Dosing after preculture 24h in saturated humidity incubator.Continue to cultivate 24th, after 48h, take in 0.4% trypan blue solution 0.15ml addition 0.6ml cell suspensions, fully mix, drawn less after dyeing 5min Amount mixed liquor, injection blood cell counting plate cell, number living cells, is repeated 3 times under light microscope, takes its average.Ask its thin Born of the same parents' appreciation rate (%)=(T-T₀)/(C-T₀)×100.Wherein C represents the cell number of control group；T represents the cell number of dosing group； T₀Represent cell number during control flat board measure dosing.If the final cell number of dosing group is more than T₀, illustrate cell after dosing Still grow.If the final cell number of dosing group is less than T₀, cell is killed after illustrating dosing.Obtained using regression equation IC₅₀.From the experimental results, the final cell number of dosing group is less than T₀, then illustrate that medicine cell growth has inhibitory action, plus Cell is killed after medicine.IC₅₀Value is as shown in the table：

	Cis-platinum	The compounds of this invention	Berberine	Melbine
					IC₅₀(μmol/L)	20.54±2.63	50.28±3.91	53.25±8.44	61.47±4.76

3rd, damage of the compound of the present invention to K562 cell DNAs

DNA damage degree determines cell life and death to a certain extent, its detection method SCSE technologies.This experiment uses this Technology (specific method does not elaborate herein with general detection DNA Damage method), detection berberine is to K562 cells The damaging action of DNA.By the K562 cells of exponential phase with 10 × 10⁴·ml^-1Concentration be inoculated in 24 well culture plates, put In 37 DEG C, 5%CO₂After dosing continues to cultivate 48h after preculture 24h in saturated humidity incubator, cell (1000r is collected min^-1Centrifugation 5min, removes supernatant).Washed twice with cold PBS, plus PBS is mixed and is made cell suspension.

4th, compound induction K562 Apoptosis of the present invention

Principle：Acridine orange (AO) can pass through the complete cell of after birth, and embedded nucleus DNA is allowed to the green glow of emitting bright. Ethidium bromide (EB) is only capable of the cell being damaged through after birth, and embedded core DNA sends out Chinese red fluorescence.In fluorescence microscopy Microscopic observation, can See four kinds of cellular morphologies：Living cells (V) nuclear chromatin fluoresced green, morphosis is normal；Viable apoptotic cell (EA) core contaminates Chromaticness jaundice green fluorescence, in pyknosis shape or round bead shape；Non-viable apoptotic cell (LA) nuclear chromatin is Chinese red or orange, structure It is pyknosis shape or round bead shape；Non-viable non-apoptotic cell (N) nuclear chromatin sends out Chinese red fluorescence, and structure is normal.

Method：Take the logarithm the K562 cells in growth period, with 5 × 10⁴·ml^-16 orifice plates are inoculated in, per hole 3mL, 37 is placed in DEG C, 5.0%CO₂Saturated humidity incubator in cultivate 24h after dosing.The test drug concentrations of K562 cells are respectively 40,80, 160μm·L^-1, the isometric complete medium of control group addition.After continuing to cultivate 24,48h, 1000rmin^-1Collect cell, PBS is washed and once take afterwards 100L cell suspensions and 4 μ LAO/EB dye liquors (1：1100μg·mL^-1) mix, smear, excitation light wave immediately A length of 570nm Fluirescence observations simultaneously count at least 300 cells.Calculate apoptosis rate (%)=(EA+LA)/(V+N+EA+LA) × 100%.Experiment above is repeated 3 times the above.After 24h being acted on the compounds of this invention group of various concentrations and control group to K562 Apoptosis rate as a example by carry out damaging action of each compound of comparing to K562 cell DNAs.

Experimental result shows, with the increase of compound concentration, K562 apoptosis rates increase, and illustrate apoptosis rate and sample Concentration is in dose-dependence.

The blood sugar reducing function effect test of the compound of the present invention of embodiment 12

With acarbose as positive control drug, external enzyme level experiment is first passed through, determine the compounds of this invention to glucose The inhibitory action of glycosides enzyme, calculates its IC₅₀Value, if the value is smaller, Ke Yikaolv carry out hypoglycemic experiment in animal body.(body herein Outer enzyme level testing program is similar to above-mentioned anti-inflammatory scheme, does not elaborate herein), internal hypoglycemic test method, with blood high Sugared rat investigates the internal blood sugar reducing function of the compounds of this invention for study subject.

Hyperglycemic rat is modeled and packet：Cleaning grade SD rats (200 ± 209) 70 is chosen, male and female half and half, balance is raised 3d, detects whole blood blood sugar, without pathoglycemia situation before modeling with blood glucose meter.Rat is divided into control group 10 at random, a modeling group 60, sub-cage rearing gives basal feed and sterile tap water during experiment.Water is can't help in 12h fasting before modeling, in tail vein Injection alloxan (6omg.kg^-1Body weight), control group is injected with the physiological saline of dosage.5 days rear side blood sugar of Continuous Observation, choosing Fasting blood-glucose is selected in 13~20mmolL^-1Between test use.

Selection models successfully diabetes rat 40, is randomly divided into 4 groups, every group 10, set up respectively hyperglycemia model group, Jamaicin group, the compounds of this invention group and positive control (melbine) group, are marked, by group with 0.5% picric acid solution respectively Other sub-cage rearing, male and female are separate.

Administration：2% Tween-80 solution prepares administrable jamaicin, the compounds of this invention and melbine solution, and concentration is 10mg.ml^-1.Administration group rat presses 100mg.kg^-1Body weight dose gavage correspondence liquid 10mL.kg^-1, body weight, control group and model Group rat oral gavage 2% Tween-80 solution 10mL.kg^-1Body weight, daily gastric infusion 1 time, continuous gavage 15 days.Zoopery exists Standart animal test room is carried out, 23 ± 2 DEG C of room temperature, humidity 50~60%, and room lighting and dark replace 12h/d, and water 1 is changed daily It is secondary, change within two days bedding and padding 1 time, each group freely absorbs drinking-water.

Experimental rat blood sugar detection and function of blood sugar reduction evaluation：On the day of each administration group gastric infusion and the 5th after administration, 10th, 15 days tail veins take blood on an empty stomach, and each group rat fasting blood-glucose value is determined with blood glucose meter, compare each group rat fasting blood-glucose value and Blood sugar declines percentage.Blood sugar decline percentage=(blood glucose value after blood glucose value-experiment before experiment) preceding blood glucose value of/experiment × 100%.

Statistical procedures:Statistical procedures are carried out to data using SPSS13.0 statistical softwares, all continuous datas are with " Number+standard deviation (x ± s) " represents that standard deviation is calculated by " n-1 " method, the comparing of multigroup sample average, using single factor test Variance analysis, P<0.05 judges that difference has statistical significance, P<0.01 judges that difference has significant statistical significance.

By calculate analyze, the compounds of this invention compared with berberine and melbine, P<0.05, it is statistically significant, It is variant, illustrate that the hypoglycemic effect of the compounds of this invention is better than alone berberine and melbine.

The effect for reducing fat effect test of the compound of the present invention of embodiment 13

Fat Golden Hamster with high lipid food induction after giving the compounds of this invention, detects body weight, liver as study subject Weight, the index such as fat body ratio and blood fat by observing its fat-reducing effect, and then inquires into its effect for reducing fat.

(1) material

Instrument：Supercentrifuge, normal rat experiment cage, electronic scale；

Medicine and reagent：The compounds of this invention, Li Sita difficult to understand, triglyceride reagent box, HDL-C kits, LDL-C reagents Box.

Animal：Male golden yellow gopher 60,

(2) experimental technique

Animal model and packet：After 60 male golden yellow gophers give basal feed adaptation nursing 3 days, 10 are randomly selected As Normal group, remaining its free choice feeding high lipid food (composition that allows：10% lard, 10% yolk powder, 1% cholesterol, 79% basal feed).After feeding four weeks, water 12h is can't help in empty stomach fasting, extracting vein blood after eye socket, determines its body weight, TC, TG, LDL-C and HDL-C.The successful Golden Hamster of modeling is randomly divided into 5 groups (n=10)：Model group, orlistat group, the present invention The basic, normal, high dosage group of compound, pharmaceutical intervention 4 weeks, Normal group and the isometric physiological saline of the daily gavage of model group, it is difficult to understand Li Sita groups administration 42mg/kg daily, the basic, normal, high dosage of the compounds of this invention respectively by daily 23.35,46.70, 70.05mg/kg is administered, and Golden Hamster is freely ingested, and is drunk water；The illumination rhythm and pace of moving things 12L, 12D (7:00-19:00), room temperature (24 ± 2) DEG C, humidity：(55 ± 10) %.

(3) index determining

1st, body weight, body appetite long and surplus are determined：During testing, each group animal takes single cage to raise and freely ingest, Drinking-water.A body weight (being weighed before gastric infusion) and surplus appetite are determined, and records result within every 3 days.Surveyed under narcosis before putting to death Fixed its body (length of nose to anus) long, waistline, by formula [Lee, s=(body weight)^-(1/3)×10³/ body is (cm) long] calculate Lee, s index.

2nd, serum TC, TG, LDL-C, HDL-C are determined：Before taking blood, water 12h is can't help in empty stomach fasting, with capillary glass tube in eye Vena orbitalis posterior takes blood, and after fasting 2h, 5000r/min centrifugation 10min take supernatant, and -20 DEG C of preservations are surveyed using biochemical reagents box Determine TC, TG, LDL-C, HDL-C in serum.

3rd, liver weight, testis and perirenal fat are determined：Take all fat around liver, perinephric fat and testis and claim Weight, calculates the ratio (fat body ratio) of fat/body weight.

(4) data processing

Statistical analysis is carried out to the data obtained with SPSS17.0 softwares, data is calculated and is represented with " mean+standard deviation ", entered Compare between row one-way analysis of variance, group using t inspections, P<0.05 is that difference is statistically significant, P<0.01 has aobvious for difference Write statistical significance.With the compounds of this invention group after medication four weeks and positive control make hyperlipemia group mouse TC, TG, LDL-C, The effect for reducing fat of the medicine is tentatively probed into as a example by the percentage that HDL-C declines.

The compounds of this invention (P compared with berberine and curcumin<0.05), there is significant difference, and three is right with the positive (P is compared according to medicine Austria Li Sita<0.01), there is the effect for reducing fat of significant significant difference, i.e. the compounds of this invention better than alone Berberine or melbine, but not as good as positive control drug orlistat.

The compound of the present invention of embodiment 14 is tested the action effect of cardiovascular and cerebrovascular disease

It is (main by seeing by inquiring into effect of the compounds of this invention to APOE (-/-) mouse early atherosclerosis Examine the influence to mouse RCCA adventitia removal, vascular collagen and the elastic plate) the compounds of this invention is studied in the heart Effect in terms of cranial vascular disease.

(1) materials and methods

1st, instrument and equipment

(1) the rotary automatic dehydrators of TISSUE-TEKII:Someiyoshine company

(2) TISSUE-TEK, TECS tissue embedding machine:Someiyoshine company

(3) freezing microtome:German Leica companies

(4) ultra low temperature freezer:SANYO GS company

(5) ordinary optical microscope:Japanese Nikon companies

(6) polarization microscope:Japanese Olympus companies

(7) fluorescence microscope:Japanese Nikon companies

(8) ImagePro-Plus6.0 image analysis softwares:MacroMedia companies of the U.S.

(9) data acquisition and analysis system SPSS17.0:PowerLab companies of the U.S.

(10) 7600 automatic analyzers:HIT

2nd, medicine and reagent

(1) the compounds of this invention (self-control)；

(2) Atorvastatin：Hangzhou Mo Shadong pharmaceutical Co. Ltds；

(3) normal saline solution：Beijing Co., Ltd of Double-Crane Pharmaceutical Co., Ltd；

(4) amobarbital sodium：Chemical Reagent Co., Ltd., Sinopharm Group；

(5) Hematoxylin-eosin biological stain：Beijing Chemical Plant；

(6) phosphate buffer (phosphate-buffered saline, PBS, 0.01M, pH7.4)：Weigh 8gNaCl、0.2gKCl、1.44gNa₂HPO₄And 0.24gKH₂PO₄, be dissolved in 800ml distilled water, with HCI adjust solution ph to 7.4, finally add distilled water to be dissolved to IL；

(7) 4% paraformaldehyde liquid:409 paraformaldehydes are weighed, is placed in flask, add 800ml 0.l mol/L phosphoric acid to delay Fliud flushing, is heated to 60 DEG C, and magnetic agitation is completely dissolved powder, finally supplies the PB to 1000ml of 0.lmol/L, fully mixes；

(8) picrosirius staining liquid：0.1g sirius reds, are dissolved in 100ml picric acid saturated solutions.

(9) Victoria blue dyeing liquor：Victoria blue 2g, dextrin 0.50g, resorcinol 4g, distilled water 200mL will Above-mentioned mixing is boiled in 5min, then the 30% ferric chloride solution addition upper liquid that will have been boiled, and continues to boil 2min, is now constantly stirred Mix in colloidal, after the cold filtration that reduces internal heat, the residue on paper is placed in 60 DEG C of insulating boxs and is dried, finally dry residue is dissolved in In 75% alcohol of 500ml, concentrated hydrochloric acid 5ml and phenol 5g is added, put room temperature standby；

(10) goat anti-rabbit igg of rhodamine mark:Beijing biotech company of Zhong Shan Golden Bridge；

(11) the anti-quencher of fluorescence:Beijing biotech company of Zhong Shan Golden Bridge.

(2) experimental animal and packet

6 week old male APoE (-/-) mouse 20 with C57BL/6 as genetic background, is purchased from Department Of Medicine, Peking University.Raise Support in the animal housing of Sino-Japanese Friendship Hospital for meeting secondary animal feeding standard.Indoor purifying air stream is moved Gas, keeps 22 DEG C~25 DEG C of room temperature, humidity 50% or so, 12 hours light and shade cycles (7am-7Pm).Mouse feeder is in standard mouse cage In, mouse cage, drinking bottle, bedding and padding are through autoclave sterilization.Mouse free water, adaptability feed 6 days after, raise with containing lard 21%, The high lipid food (60 bore sterilizing treatment with irradiation) of cholesterol 1.25% and common mixed feed meal 77.75%.Experimental animal operates Process is strictly carried out in accordance with the code of Beijing Union Medical College and Ethics Committee of China-Japan Friendship Hospital, and the treatment to animal meets Animal protection ethics.Mouse is randomly divided into 4 groups, the large, medium and small dosage group of blank control group, melbine, every group each 5 Only, influence of the various dose group to ApoE (-/-) mouse early atherosclerosis after observation administration.

Medication：According to《Pharmacological experimental methodology》(Xu Shuyun is edited, the third edition in 2002, People's Health Publisher) In people and mouse dosage reduction formula, Experimental agents dosage is converted by body weight.According to being grown up, daily medication is faced The common dose that bed is recommended, mouse consumption is converted to by adult with the body weight conversion factor 9.01 of mouse, and Atorvastatin group is given Atorvastatin 3mg/kg/d is given, blank the compounds of this invention group gives physiological saline 0.2ml/kg/d, the compounds of this invention group The heavy dose of group 240mg/kg/d of the compounds of this invention, middle dose group 160mg/kg/d, small dose group 80mg/kg/d are given with gavage Mode be administered, continue 4 weeks.

1st, influence of the compounds of this invention to Ap0E (-/-) mouse early atherosclerosis RCCA adventitia removal

Materials and section

(1) draw materials：After experiment mice fasting 12h, with 1% amobarbital sodium intraperitoneal injection of anesthesia mouse, lain on the back, Head and four limbs are fixed on operating table, and after alcohol disinfecting, longitudinal incision abdomen chest and skin of neck are successively separated, and cut off tabula, Exposure heart, after inferior caval vein takes blood, is irrigated, after blood is rinsed well, cruelly with 0.9% physiological saline by left ventricle Dew RCCA sheath, dissociate RCCA, is about 1cm, is rinsed with ice physiological saline, after filter paper is blotted, is placed on On the small circular hardboard of the one a diameter of 3cm for scribbling OCT frozen section embedding mediums, and OC fourths are coated on whole sample, put Solid-state is frozen into the small beaker of isopentane in filling for existing Liquid nitrogen precooler about 1min, after be placed in -80 DEG C of refrigerators and preserve standby With.

(2) cut into slices:It is continuously crosscutting with 6 μm of thickness since RCCA near-end lower section, leave and take 1 for every 50 μm and cut Piece, is placed on slide, and every mouse takes 8 sections.

Pathological examination and analysis:Frozen section row Hematoxylin-eosin (HE) is dyeed, around high power Microscopic observation blood vessel The Infiltrating of inflammatory cell.Range of observation is the interstitial tissue of mouse RCCA and surrounding, by dyeed tangent plane, The length of vessel that appears is different, it was observed that area it is also different, sample inflammatory cell distribution density degree has very big difference in addition Not, it is unified standard, we choose each intensive region of observed vasculitis cell distribution, and every section is in LEICA The random statistical number for calculating 8 400 times of object lens visual field positive cells sums as the sample on Image analysis system Value.

Frozen section HE staining procedures：

1. frozen section, recovers 20min at room temperature；

2. cut into slices into HarriS haematoxylins 15min；

3. flowing water rinses 5min；

4. 0.5% hydrochloride alcohol breaks up 30s；

5. flowing water rinses 5min；

6. eosin stains 10min；

7. flowing water rinses 5min；

8. gradient alcohol dehydration；

9. transparent 15min × 2 time of dimethylbenzene, middle victory mounting.

Statistical procedures：Using SPSSl7.0 softwares, dosage information is represented using mean ± standard deviation (x ± S), multigroup ratio Relatively using one-way analysis of variance (one-way ANOVA), compare two-by-two between group and use LSD, P<0.05 has statistics for difference Meaning.During with heavy dose group, the compounds of this invention group and positive control drug are to mouse RCCA adventitia removal number of cells Influence compare influence of the medicine to Ap0E (-/-) mouse early atherosclerosis RCCA adventitia removal, enter And inquire into its effect in terms of cardiovascular and cerebrovascular disease.

Group	Atorvastatin	The compounds of this invention	Berberine	Melbine
					Inflammatory cell number (individual)	5±2	25±3	29±3	35±2

Result shows：The compounds of this invention (P compared with berberine and melbine<0.05), there is significant difference, show The compounds of this invention is better than berberine or melbine to the inhibitory action of mouse RCCA adventitia removal.

2nd, influence of the compounds of this invention to Ap0E (-/-) mouse early atherosclerosis vascular collagen and elastic plate

Experimental animal, feeds packet, and materials always moved with to Ap0E (-/-) right neck of mouse early atherosclerosis with being cut into slices The experiment of arteries and veins adventitia removal is identical.

Pathological examination and analysis：The ruthless scarlet resisdye Victoria blue dyeing of frozen section day, frozen section sirius red Dyeing is chosen section and is dyeed with 0.1% picric acid sirius red dye liquor, to assess I types and 111 Collagen Type VIs.Every mouse leaves and takes 8 Section is opened, the collagen content average values of this 8 sections represent every RCCA collagen content value of mouse.

Picrosirius staining step：

(1) 20min is recovered under frozen section room temperature；

(2) dimethylbenzene and absolute alcohol (1 are moved into:1) 5min or so in mixed liquor；

(3) 100%, 95%, 85%, 70% alcohol is entered, at different levels is 5min；

(4) distillation washing 3 times；

(5) 60min is contaminated with picric acid sirius red dye liquor；

(6) absolute alcohol directly breaks up and dehydration；

(7) dimethylbenzene is transparent, neutral gum mounting.

Frozen section Victoria blue is dyeed：Choose section to be dyeed with Victoria blue dye liquor, to assess elastic fibers point Level and Internal-media thickness.Every mouse leaves and takes 8 sections, and this 8 elastic fibers of section are classified and its media thickness average value Represent the classification of RCCA elastic fibers and the value of Internal-media thickness of every mouse.

Victoria blue staining procedure：

Recover 20min under (l) frozen section room temperature；

(2) to cut into slices and wash 1min in 75% alcohol；

(3) 30min or so in Victoria blue dyeing liquor；

(4) directly with 95% alcohol color separation several seconds；

(5) with distillation washing three times,

(6) dimethylbenzene is transparent, neutral gum sealing.

Statistical procedures：Using SPSSl7.0 softwares, dosage information is represented using mean ± standard deviation (x ± S), multigroup ratio Relatively using one-way analysis of variance (one-way ANOVA), compare two-by-two between group and use LSD, P<0.05 has statistics for difference Meaning.During with heavy dose group, the compounds of this invention group and positive controls are to the type of mouse I, III Collagen Type VI content (%) and elastic force The influence of the sick rate (%) of I grade of fiber investigates the medicine to Ap0E (-/-) mouse early atherosclerosis vascular collagen and bullet The influence of power plate.

Group	Atorvastatin	The compounds of this invention	Berberine	Melbine
					NTx content (%)	21.52±1.63	49.24±2.16	59.42±3.17	65.43±1.70
III Collagen Type VI content (%)	4.53±2.31	39.51±2.82	42.35±1.53	49.35±2.72
					I grade of disease (%) of elastic fibers	60.48±1.52	26.36±2.18	14.30±1.85	12.65±2.34

Result shows：With I type, III Collagen Type VI content (%) and the sick rate (%) of I grade of elastic fibers for control parameters, the present invention Compound compares (P with berberine and melbine<0.05), there is significant difference, and three compares (P with positive controls< 0.01), there is a significant significant difference, and then show the compounds of this invention to Ap0E (-/-) mouse early atherosclerosis The influence of vascular collagen and elastic plate is better than alone berberine or curcumin, but not as good as positive control drug.

It is last to should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than the present invention is protected The limitation of scope is protected, although being explained in detail to the present invention with reference to preferred embodiment, one of ordinary skill in the art should Understand, technical scheme can be modified or equivalent, without deviating from the essence of technical solution of the present invention And scope.

Claims

1. a kind of compound, it is characterised in that the structural formula of the compound is：

2. a kind of preparation method of compound as claimed in claim 1, it is characterised in that the described method comprises the following steps：

(1) compound shown in formula (1) and bromo agent are carried out into bromo-reaction in solvent orange 2 A, obtains compound shown in formula (2)；

(2) formula (2) compound that will be obtained in step (1) in the presence of base catalyst, with compound shown in formula (3) in solvent There is ring-closure reaction in B, that is, obtain formula of the present invention (4) compound.

3. the preparation method of compound as claimed in claim 2, it is characterised in that bromo agent is bromoacetic acid in the step (1) In methyl esters, bromoacetate, bromoacetic acid butyl ester, methyl chloroacetate, ethyl chloroacetate, butyl chloroacetate, iodoacetic acid methyl esters extremely Few one kind；Solvent orange 2 A is acetonitrile, DMF, methyl alcohol, acetone, ethanol, dimethyl sulfoxide (DMSO), tetrahydrofuran, dichloromethane, dimethyl Asia At least one in sulfone, dioxane.

4. the preparation method of compound as claimed in claim 2, it is characterised in that base catalyst is methyl alcohol in the step (2) At least one in sodium, caustic alcohol, metallic sodium；Solvent B is acetonitrile, DMF, ethyl acetate, methyl alcohol, acetone, pyridine, quinoline, two At least one in methyl sulfoxide, tetrahydrofuran, dichloromethane, dimethyl sulfoxide (DMSO), dioxane.

5. the preparation method of compound as claimed in claim 2, it is characterised in that solvent orange 2 A is changed with formula (1) in the step (1) The mol ratio of compound is 5:1~20:1；Bromo agent is 0.8 with the mol ratio of formula (1) compound:1~3:1.

6. the preparation method of compound as claimed in claim 2, it is characterised in that base catalyst and formula (2) in the step (2) The mol ratio of compound is 0.04:1~5:1；Solvent B is 5 with the mol ratio of formula (2) compound:1~20:1；Formula (3) compound It is 0.5 with the mol ratio of formula (2) compound:1~5:1.

7. the preparation method of compound as claimed in claim 2, it is characterised in that bromo-reaction temperature is in the step (1) 30~80 DEG C, the reaction time is 3~12 hours.

8. the preparation method of compound as claimed in claim 2, it is characterised in that the temperature of ring-closure reaction in the step (2) It it is 40~100 DEG C, the reaction time is 3~20 hours.

9. the preparation method of compound as claimed in claim 2, it is characterised in that step (1) Chinese style (1) compound is used Following methods are prepared from：By compound shown in formula (5) in solvent C at 120~170 DEG C react, make its 9 methyl take off Go, obtain compound shown in formula (1)；Wherein described solvent C is DMF, DMAC, DMSO, HMPT, HEPT, NMP, diphenyl ether, pyrrole One kind in pyridine, quinoline, the solvent C is 5 with the mol ratio of compound shown in formula (5):1~20:1；

10. a kind of compound as claimed in claim 1 is being prepared for treating tumour, cancer, hyperglycaemia, high fat of blood, heart and brain blood Purposes in the medicine of pipe disease.