CN105566353A

CN105566353A - Compound and preparation method thereof

Info

Publication number: CN105566353A
Application number: CN201610021313.4A
Authority: CN
Inventors: 李卫民; 冯毅凡; 朱贺年; 李铀; 张志朋
Original assignee: Beijing Yishengtang Medicine Science And Technology Research Co Ltd
Current assignee: Beijing Yishengtang Medicine Science And Technology Research Co Ltd
Priority date: 2016-01-13
Filing date: 2016-01-13
Publication date: 2016-05-11

Abstract

The invention discloses a compound. The compound is prepared from berberrubine and magnolol through two times of nucleophilic substitution reactions under certain conditions. According to the compound, the inventor is guided by the traditional Chinese medicine pair theory and the traditional Chinese medicine clinical medication experience, berberrubine and magnolol are purposefully and selectively combined through a specific synthetic method, and the obtained compound has the effects of increasing the effect and reducing toxins, and has better effects on the aspects of inflammatory resisting, tumor resisting, cancer resisting, hyperglycemia and hyperlipidemia reducing, bacterium resisting, cardiovascular and cerebrovascular disease resisting and the like. Meanwhile, the invention discloses a preparation method of the compound and application of the compound in preparing medicine for treating tumors, cancer, hyperglycemia, hyperlipidemia and the cardiovascular and cerebrovascular diseases.

Description

A kind of compound and preparation method thereof

Technical field

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

Background technology

Now, along with the development of modern pharmacology, molecular biology scheduling theory and related science technology, the development approach of natural drug and means are also being constantly brought forth new ideas.At present, the approach of developing new drug roughly has three, i.e. Folk medicine approach, experience medicine approach and molecular pathways.Wherein along with the development of molecular biology, genetics and the information processing technology etc., molecular pathways becomes the main path of developing new drug gradually.But based on the current national conditions of China, and due to the limitation of technical equipment, current Folk medicine approach, experience medicine approach are the main paties of China's developing new drug, and these two kinds of approach are based on medicinal plant.

Take medicinal plant as the medicine that fundamental development gets up, in history huge effect is served to the preventing and curing diseases of the mankind, rehabilitation and fertility procreation.Natural drug available sources is a lot of, mostly derives from plant; According to estimates, about there are 40 ~ 500,000 kind of plant in the whole world, but only there is few part to carry out chemical composition and active testing research thereof, China is also one of the abundantest country of plant resources, therefore fully excavate and utilize the compound of nature structure diversity to develop new natural drug, promoting that China's medical sci-tech advances at utmost speed, open up new market, promote the development of China's natural drug, meet medication economics challenge of globalization to have great importance.Medicinal plant is the important sources of developing new drug, and at present, in the new drug of whole world approval, be natural drug close to half, medicine is from natural product.Now, the population of nearly 60% is also had also to rely on plant amedica to carry out disease preventing and treating completely in the world.

But along with the development of disease, the problems such as viral resistance, body resistance, certain medicinal plant of simple dependence, or in medicinal plant, ingredient is the new drug that precursor is researched and developed, and far can not meet the needs of people to medicine.For certain specific disease, people often will by two or more, even more medicine is taken simultaneously and just can be played certain therapeutic action.

The TCM Culture of China has the history of 5000, drug matching, and Chinese medicines is to theory our rarity especially.The synthesis of Chinese medicines to theoretical direction compound is important Folk medicine approach, the experience medicine approach of research, developing new drug.

" medicine to " is the minimum unit of Chinese medicine preparation compatibility and relatively fixes combination and the compatibility application of two taste Chinese medicines, be the drug matching application regularity that ancient Chinese medicine doctor is summed up in Long-term clinical medical practice, its central role is improvement, strengthens clinical efficacy or reduce toxicity, side effect.Traditional sense right research of adding medicine to carries out for the amount of two taste medicinal materials, and single medicinal material is individual multicomponent large compound inherently, and the characteristic of Chinese medicine is multicomponent, Mutiple Targets, synergy.In recent years right to medicine compatibility research, discloses multicomponent between medicine pair influencing each other, explores the mechanism of action of its synergy, attenuation; The basic substance for the treatment of by Chinese herbs onset is the effective constituent contained by it, and influencing each other between this effective constituent is exactly the marrow of medicine to research.

From 1998, we carried out new drug development to the ancient prescription " bark of official magnolia ball " in Holy Benevolent Prescriptions, obtained the New Drug Certificate (traditional Chinese medicines card word Z20100004) of " yellow thick antidiarrheal dripping pill " to 2010 and produce official written reply (2010S00384), the essence of research discovery " yellow thick antidiarrheal dripping pill " prescription compatibility is exactly the right mutual compatible combination of " coptis-bark of official magnolia ", " coptis-banksia rose ", " bark of official magnolia-rhizoma zingiberis ", " coptis-rhizoma zingiberis " these medicines; Wherein most crucial in prescription medicine is to being " coptis-bark of official magnolia " medicine pair; The most crucial effective constituent of this medicine centering is berberine Honokiol, cause the design that medication is synthesized synergy, this two compound of attenuation theoretical direction berberine Honokiol or spliced thus, the result of synergy, attenuation after attempting such splicing, can have been played; Can by the berberine in the coptis and the magnolol in the bark of official magnolia with different parent nucleus to become than being stitched together one new can synergy, attenuation " multinuclear molecule ", multinuclear molecular compound D in this patent is under this theoretical direction, object is had to obtain bimolecular berberine and monomolecular magnolol, optionally be stitched together, form a brand-new compound.

Summary of the invention

The object of the invention is to based on above-mentioned technical background and a kind of new compound is provided, described compound is by berberrubine Honokiol by specific means, formed than being stitched together with different parent nucleus, and it has better anti-inflammatory, antitumor anticancer, hypoglycemic, the effect such as antibacterial.Meanwhile, the present invention also provides the Preparation method and use of described compound.

For achieving the above object, the technical scheme that the present invention takes is: a kind of compound, and its structural formula is:

Compound of the present invention, berberrubine Honokiol is adopted to be raw material, form through special Reactive Synthesis, containing bimolecular berberrubine and monomolecular magnolol in described compound, form three core molecule structures of particular configuration, the existence of methoxyl group in described molecule, makes to have effect that is anti-oxidant, scavenging free radicals in the compounds of this invention, thus plays the effect delayed senility.In in molecule, allylic existence in magnolol parent nucleus, makes the compounds of this invention have the effect of nervus centralis, the existence of quaternary ammonium (Nitrogen ion) in berberrubine parent nucleus, makes the compounds of this invention have effect that is antitumor, hypoglycemic.

The compounds of this invention adopts berberrubine Honokiol to be raw material, and berberrubine described in practice can prepare gained by the Berberine in berberine, also directly can be purchased from market; Described magnolol derives from the bark of official magnolia.Widely used a herb during the described bark of official magnolia, clinically multiplexly treat bacteriological infection inflammation and gastrointestinal tract disease etc., its main component is magnolol and isomer thereof, there is many-sided pharmacological action, nervus centralis suppression of flaccid muscles if any central, anti-inflammatory, antibacterial, antiulcer agent, anti-oxidant, antitumor, regulating effect etc. rapidly.Berberrubine and magnolol combine by the present invention, and by specific mole when specific reaction conditions be prepared from, described compound has specific three core molecule structures, can significantly reduce on the basis of berberine Berberine toxicity, more can play the effect of efficacy enhancing and toxicity reducing, in anti-inflammatory, antitumor, hypoglycemic, antibacterial and cardiovascular and cerebrovascular diseases etc., there is better effect.

In addition, present invention also offers a kind of preparation method of compound described above, described preparation method comprises the following steps:

(1) compound of formula (1) and bromo agent are carried out bromo-reaction in solvent orange 2 A, obtain the compound of formula (2);

(2) compound of the formula (2) step (1) obtained is under the acting in conjunction of alkaline catalysts A and phase-transfer catalyst A, in solvent B, there is nucleophilic substitution reaction with the compound of formula (3), obtain the compound of formula (4);

(3) compound of the formula (4) step (2) obtained is in the acting in conjunction of alkaline catalysts B and phase-transfer catalyst B, in solvent C, secondary nucleophilic substitution reaction is there is with the compound of formula (2), obtain the compound of formula (5), be the compounds of this invention;

The preparation method of compound of the present invention, by specific method, the synthesis of the berberrubine Honokiol parent nucleus of specified proportion is stitched together, thus form a kind of three core molecule structures with ad hoc structure, this splicing is not blindly, but at Chinese medicines under the guidance of theory, tcm clinical medication experience, have object, selectively the two combined according to specific ratio, find form three core molecule structures of ad hoc structure compound possess the effect of efficacy enhancing and toxicity reducing.Preparation method of the present invention is simple to operate, can rapidly and efficiently obtain compound of the present invention.

As the preferred implementation of the preparation method of compound of the present invention, bromo agent in described step (1) is 1,2-ethylene dibromide, 1,2-ethylidene periodide, 1,2-ethylene dichloride, 1,2-dibromobutane, 1, at least one in 2-bis-butyl iodide, 1,2-propylene dichloride, 1,2-dibromo-hexane.As the more preferably embodiment of the preparation method of compound of the present invention, the bromo agent in described step (1) is glycol dibromide.When glycol dibromide is selected in described bromo agent, described 1, two carbon in 2-ethylene dibromide molecule both can overcome intermolecular resistance, ensure the generation of nucleophilic substitution reaction, maintain again the stability of synthesized compound, avoid it unstable in vivo and decompose.

As the preferred implementation of the preparation method of compound of the present invention, alkaline catalysts A in described step (2) and step (3) and alkaline catalysts B is independent separately, be sodium hydroxide, lithium hydroxide, potassium hydroxide, salt of wormwood, sodium carbonate, sodium bicarbonate, saleratus respectively, triethylamine, at least one in DIPEA.As the more preferably embodiment of the preparation method of compound of the present invention, the alkaline catalysts A in described step (2) and step (3) is sodium carbonate, described alkaline catalysts B is salt of wormwood.When described alkaline catalysts A be sodium carbonate, described alkaline catalysts B be salt of wormwood time, its alkalescence is moderate, and present inventor is found by large quantity research, when described alkaline catalysts A be sodium carbonate, described alkaline catalysts B be salt of wormwood time, the yield of product is apparently higher than other alkaline catalystss.

As the preferred implementation of the preparation method of compound of the present invention, in described step (2), the molar weight of alkaline catalysts A molar weight and formula (2) compound is than being 0.1:1 ~ 50:1, and in described step (3), the molar weight of alkaline catalysts B and the molar weight of formula (4) compound are than being 0.1:1 ~ 20:1.

As the preferred implementation of the preparation method of compound of the present invention, phase-transfer catalyst A in described step (2) and step (3) and phase-transfer catalyst B is independent separately, be at least one in Tetrabutyl amonium bromide, tetrabutyl ammonium fluoride, TBAH, 4-butyl ammonium hydrogen sulfate, three (3,6-dioxaheptyl) amine, benzyl triethyl ammonium bromide, tri-n-octyl methyl ammonium chloride respectively.As the more preferably embodiment of the preparation method of compound of the present invention, the phase-transfer catalyst A in described step (2) and step (3) and phase-transfer catalyst B is Tetrabutyl amonium bromide.Present inventor finds through large quantity research, and when Tetrabutyl amonium bromide selected by described phase-transfer catalyst, its catalytic effect is obviously better than other phase-transfer catalyst, and Tetrabutyl amonium bromide is cheap and easy to get.

As the preferred implementation of the preparation method of compound of the present invention, in described step (1), the temperature of bromo-reaction is 40 ~ 100 DEG C, and the reaction times is 3 ~ 15 hours; In described step (2), the temperature of nucleophilic substitution reaction is 40 ~ 100 DEG C, and the reaction times is 3 ~ 15 hours; In described step (3), the temperature of secondary nucleophilic substitution reaction is 20 ~ 80 DEG C, and the reaction times is 5 ~ 24 hours.As the preferred implementation of the preparation method of compound of the present invention, in described step (1), bromo-reaction temperature is 70 ~ 80 DEG C, and the reaction times is 8 ~ 10 hours; In described step (2), the temperature of nucleophilic substitution reaction is 70 ~ 80 DEG C, and the reaction times is 8 ~ 10 hours.Present inventor finds through research, when the temperature of the nucleophilic substitution reaction in the bromo-reaction in described step (1) and step (2) be 70 ~ 80 DEG C, the reaction times is 8 ~ 10 constantly little, can yield be ensured, there is again good economy.

As the preferred implementation of the preparation method of compound of the present invention, in described step (1), the mol ratio of solvent orange 2 A and formula (1) compound is 5 ~ 10:1, and described solvent orange 2 A is at least one in acetonitrile, DMF, methyl alcohol, acetone, ethanol, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), methylene dichloride, dimethyl sulfoxide (DMSO), dioxane; In described step (2), the mol ratio of solvent B and formula (2) compound is 5 ~ 10:1, and described solvent B is at least one in acetonitrile, DMF, ethyl acetate, methyl alcohol, acetone, pyridine, quinoline, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), methylene dichloride, dimethyl sulfoxide (DMSO), dioxane; In described step (3), the mol ratio of solvent C and formula (4) compound is 5 ~ 10:1, and described solvent C is at least one in acetonitrile, DMF, ethyl acetate, methyl alcohol, acetone, pyridine, quinoline, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), methylene dichloride, dimethyl sulfoxide (DMSO), dioxane.As the more preferably embodiment of the preparation method of compound of the present invention, described solvent orange 2 A is acetonitrile, and described solvent B is acetonitrile, and described solvent C is DMF.When described solvent orange 2 A selects acetonitrile, the reactant solubility that acetonitrile reacts this step is better, and viscosity is little, and boiling point is low, and polarity is moderate, under the prerequisite that both ensure that product yield, is easy to aftertreatment again, recycling design.When described solvent B selection acetonitrile and solvent C selection DMF, there is better effect same as above.

As the preferred implementation of the preparation method of compound of the present invention, described step (1) is: add in clean there-necked flask by the compound of formula (1), add solvent orange 2 A, heating, slowly add bromo agent, temperature rises to 40-100 DEG C, back flow reaction 3 ~ 15 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treats that temperature is down to 50 ~ 60 DEG C, crystallization, then suction filtration, washing, drying, obtains the compound of formula (2).As the more preferably embodiment of the preparation method of compound of the present invention, described crystallization can adopt and add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, then carries out suction filtration, washing, drying.

As the preferred implementation of the preparation method of compound of the present invention, described step (2) is: formula (2) compound step (1) obtained adds in clean there-necked flask, add solvent B and alkaline catalysts A, stir, then compound and the phase-transfer catalyst A of formula (3) is added, react 3 ~ 15 hours at temperature is 40 ~ 100 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4).

As the preferred implementation of the preparation method of compound of the present invention, described step (3) is: the compound of the formula (4) step (2) obtained adds in clean there-necked flask, add solvent C, the compound of formula (2) is added after stirring, continue to stir, slowly add alkaline catalysts B, add phase-transfer catalyst B again, react 5 ~ 24 hours at 20 ~ 80 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5).As the more preferably embodiment of the preparation method of compound of the present invention, the compound of the formula (5) obtained is refined through further, obtains the compounds of this invention that purity is higher.Described refining further can adopt the methods such as extraction, backflow, recrystallization.

As the preferred implementation of the preparation method of compound of the present invention, described step (1) Chinese style (1) compound adopts following methods to be prepared from: reacted at 50 ~ 300 DEG C in solvent D by formula (6) compound, its 9 methyl are sloughed, obtains the compound of formula (1);

The mol ratio of described solvent D and formula (6) compound is 5 ~ 20:1; Described solvent D is the one in DMF, DMAC, DMSO, HMPT, HEPT, NMP, phenyl ether, pyridine, quinoline.

As the more preferably embodiment of the preparation method of compound of the present invention, described step (1) Chinese style (1) compound adopts following methods to be prepared from: there-necked flask formula (6) compound being placed in dried and clean, add solvent D, heating reflux reaction in electricity cover, react at temperature is 50 ~ 400 DEG C, with HPLC monitoring in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1).

As the preferred implementation of the preparation method of compound of the present invention, described step (1) Chinese style (1) compound is directly purchased from market.

Finally, present invention also offers the purposes of compound described above in the medicine for the preparation for the treatment of tumour, cancer, hyperglycemia, hyperlipidemia, cardiovascular and cerebrovascular diseases, compound of the present invention can be used for the medicine preparing treatment tumour, cancer, hyperglycemia, hyperlipidemia, cardiovascular and cerebrovascular diseases.

The compounds of this invention, contriver under the guidance of Chinese medicines to theoretical and tcm clinical medication experience, there is object, selectively by the berberrubine Honokiol of specific parent nucleus ratio by specific methods combining together, form the three core molecule structures with ad hoc structure, the compound of gained three core molecule structure has the effect of efficacy enhancing and toxicity reducing, in anti-inflammatory, antitumor, anticancer, hypoglycemic, antibacterial, resisting cardiovascular disease etc., have better effect.

The preparation method of compound of the present invention, simple to operate, rapidly and efficiently can obtain compound of the present invention.The purposes of compound of the present invention in the medicine for the preparation for the treatment of tumour, cancer, hyperglycemia, hyperlipidemia, cardiovascular and cerebrovascular diseases etc., the medicine for preparation treatment knurl, cancer, hyperglycemia, hyperlipidemia, cardiovascular and cerebrovascular diseases etc. provides new compound and selects.

Accompanying drawing explanation

Fig. 1 is the chemical structural formula of compound of the present invention.

Fig. 2 is the high resolution mass spectrum figure of compound of the present invention.

Fig. 3 is the infrared spectrogram of compound of the present invention.

Fig. 4 is the ultraviolet spectrogram of compound of the present invention.

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

Fig. 6 is the proton nmr spectra carbon spectrogram of compound of the present invention.

Embodiment

For better the object, technical solutions and advantages of the present invention being described, below in conjunction with the drawings and specific embodiments, the invention will be further described.

The structural formula of following examples formula used (1) ~ (6) compound is as follows respectively:

Embodiment 1

A kind of embodiment of the compounds of this invention, as shown in Figure 1, compound described in the present embodiment adopts following preparation method to be prepared to the chemical structural formula of compound described in the present embodiment:

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add HEPT, the mol ratio of described HEPT and formula (5) compound is 5:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 65%, and purity is 92%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add acetonitrile, the mol ratio of described acetonitrile and formula (1) compound is 5:1, heating, slowly add 1, 2-ethylene dichloride, temperature rises to 40 DEG C, back flow reaction 5 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to less than 40 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 80%, and purity is 90%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add dimethyl sulfoxide (DMSO) and triethylamine, the mol ratio of described dimethyl sulfoxide (DMSO) and formula (2) compound is 10:1, the molar weight of described triethylamine and formula (2) compound is than being 50:1, stir, then compound and the benzyl triethyl ammonium bromide of formula (3) is added, react 3 hours at temperature is 100 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 55%, and purity is 89%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add pyridine, described pyridine be 8:1 with the mol ratio of formula (4) compound, the compound of formula (2) is added after stirring, continue to stir, slowly add the mixture (mol ratio of sodium hydroxide and lithium hydroxide is 1:1) of sodium hydroxide and lithium hydroxide, the mol ratio of the mixture of described sodium hydroxide and lithium hydroxide and formula (4) compound is 10:1, add tri-n-octyl methyl ammonium chloride again, react 24 hours at 20 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.

Embodiment 2

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add DMAC, the mol ratio of described DMAC and formula (5) compound is 6:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 67%, and purity is 91%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add methyl alcohol, the mol ratio of described methyl alcohol and formula (1) compound is 6:1, heating, slowly add 1, 2-propylene dichloride, temperature rises to 100 DEG C, back flow reaction 3 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to 50-60 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 81%, and purity is 92%

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add dimethyl sulfoxide (DMSO) and sodium bicarbonate, the mol ratio of described dimethyl sulfoxide (DMSO) and formula (2) compound is 9:1, the molar weight of described sodium bicarbonate and formula (2) compound is than being 0.1:1, stir, then compound and the 4-butyl ammonium hydrogen sulfate of formula (3) is added, react 15 hours at temperature is 40 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 56%, and purity is 85%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add acetone, described acetone be 9:1 with the mol ratio of formula (4) compound, the compound of formula (2) is added after stirring, continue to stir, slowly add potassium hydroxide, the mol ratio of described potassium hydroxide and formula (4) compound is 15:1, add benzyl triethyl ammonium bromide again, react 20 hours at 25 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.Embodiment 3

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add DMSO, the mol ratio of described DMSO and formula (5) compound is 8:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 69%, and purity is 93%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add ethanol, the mol ratio of described ethanol and formula (1) compound is 10:1, heating, slowly add 1, 2-ethylidene periodide and 1, the mixture (1 of 2-bis-butyl iodide, 2-ethylidene periodide and 1, the mol ratio of 2-bis-butyl iodide is 2:1), temperature rises to 50 DEG C, back flow reaction 13 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to less than 40 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 86%, and purity is 91%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add pyridine and saleratus, the mol ratio of described pyridine and formula (2) compound is 8:1, the molar weight of described saleratus and formula (2) compound is than being 10:1, stir, then compound and the Tetrabutyl amonium bromide of formula (3) is added, react 14 hours at temperature is 50 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 58%, and purity is 87%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add quinoline, described quinoline be 5:1 with the mol ratio of formula (4) compound, the compound of formula (2) is added after stirring, continue to stir, slowly add the mixture (mol ratio of salt of wormwood and sodium carbonate is 1:2) of salt of wormwood and sodium carbonate, the mol ratio of the mixture of described salt of wormwood and sodium carbonate and formula (4) compound is 50:1, add Tetrabutyl amonium bromide again, react 18 hours at 30 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.

Embodiment 4

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add HMPT, the mol ratio of described HMPT and formula (5) compound is 10:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 71%, and purity is 96%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add the mixture (mol ratio of dimethyl sulfoxide (DMSO) and tetrahydrofuran (THF) is 1:1) of dimethyl sulfoxide (DMSO) and tetrahydrofuran (THF), the mol ratio of the mixture of described dimethyl sulfoxide (DMSO) and tetrahydrofuran (THF) and formula (1) compound is 9:1, heating, slowly add 1, 2-dibromobutane, temperature rises to 70 DEG C, back flow reaction 6 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to 50 ~ 60 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 84%, and purity is 93%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add the mixture (mol ratio of methyl alcohol and methylene dichloride is 1:1) of methyl alcohol and methylene dichloride, potassium hydroxide, the mol ratio of the mixture of described methyl alcohol and methylene dichloride and formula (2) compound is 7:1, the molar weight of described potassium hydroxide and formula (2) compound is than being 15:1, stir, then compound and the tetrabutyl ammonium fluoride of formula (3) is added, react 12 hours at temperature is 60 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 59%, and purity is 88%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add tetrahydrofuran (THF), described tetrahydrofuran (THF) be 7:1 with the mol ratio of formula (4) compound, the compound of formula (2) is added after stirring, continue to stir, slowly add N, N-diisopropylethylamine, described N, the mol ratio of N-diisopropylethylamine and formula (4) compound is 20:1, add tetrabutyl ammonium fluoride again, react 16 hours at 40 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.

Embodiment 5

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add DMF, the mol ratio of described DMF and formula (5) compound is 12:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 75%, and purity is 98%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add acetonitrile, the mol ratio of described acetonitrile and formula (1) compound is 8:1, heating, slowly add 1, 2-ethylene dibromide, temperature rises to 60 DEG C, back flow reaction 10 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to less than 40 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 88%, and purity is 95%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add acetonitrile and sodium carbonate, the mol ratio of described acetonitrile and formula (2) compound is 6:1, the molar weight of described sodium carbonate and formula (2) compound is than being 25:1, stir, then compound and the Tetrabutyl amonium bromide of formula (3) is added, react 10 hours at temperature is 80 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 60%, and purity is 90%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add DMF, described DMF be 6:1 with the mol ratio of formula (4) compound, the compound of formula (2) is added after stirring, continue to stir, slowly add salt of wormwood, the mol ratio of described salt of wormwood and formula (4) compound is 30:1, add Tetrabutyl amonium bromide again, react 15 hours at 50 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.

Embodiment 6

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add NMP, the mol ratio of described NMP and formula (5) compound is 15:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 72%, and purity is 96%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add methylene dichloride, the mol ratio of described methylene dichloride and formula (1) compound is 7:1, heating, slowly add 1, 2-dibromo-hexane, temperature rises to 90 DEG C, back flow reaction 4 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to less than 50 ~ 60 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 86%, and purity is 94%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add tetrahydrofuran (THF), the mixture (mol ratio of sodium hydroxide and sodium carbonate is 1:2) of sodium hydroxide and sodium carbonate, the mol ratio of described tetrahydrofuran (THF) and formula (2) compound is 5:1, the molar weight of the mixture of described sodium hydroxide and sodium carbonate and formula (2) compound is than being 30:1, stir, then compound and the TBAH of formula (3) is added, react 8 hours at temperature is 90 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 61%, and purity is 86%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add the mixture (mol ratio of acetonitrile and DMF is 1:1) of acetonitrile and DMF, the mixture of described acetonitrile and DMF be 10:1 with the mol ratio of formula (4) compound, the compound of formula (2) is added after stirring, continue to stir, slowly add triethylamine, the mol ratio of described triethylamine and formula (4) compound is 40:1, add TBAH again, react 12 hours at 60 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.

Embodiment 7

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add phenyl ether, the mol ratio of described phenyl ether and formula (5) compound is 16:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 70%, and purity is 97%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add the mixture (mol ratio of tetrahydrofuran (THF) and dioxane is 1:3) of tetrahydrofuran (THF) and dioxane, the mol ratio of the mixture of described tetrahydrofuran (THF) and dioxane and formula (1) compound is 8:1, heating, slowly add 1, 2-ethylidene periodide, temperature rises to 80 DEG C, back flow reaction 5 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to 50-60 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 85%, and purity is 92%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add ethyl acetate, the mixture (mol ratio of lithium hydroxide and potassium hydroxide is 2:1) of lithium hydroxide and potassium hydroxide, the mol ratio of described ethyl acetate and formula (2) compound is 8:1, the molar weight of the mixture of described lithium hydroxide and potassium hydroxide and formula (2) compound is than being 40:1, stir, then compound and the tri-n-octyl methyl ammonium chloride of formula (3) is added, react 4 hours at temperature is 100 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 63%, and purity is 84%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add the mixture (mol ratio of DMF and ethyl acetate is 1:1) of DMF and ethyl acetate, the mol ratio of the mixture of described DMF and ethyl acetate and formula (4) compound is 8:1, the compound of formula (2) is added after stirring, continue to stir, slowly add saleratus, the mol ratio of described saleratus and formula (4) compound is 45:1, add 4-butyl ammonium hydrogen sulfate again, react 10 hours at 65 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.Embodiment 8

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add pyridine, the mol ratio of described pyridine and formula (5) compound is 18:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 73%, and purity is 90%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add the mixture (mol ratio of acetonitrile and dimethyl sulfoxide (DMSO) is 1:4) of acetonitrile and dimethyl sulfoxide (DMSO), the mol ratio of the mixture of described acetonitrile and dimethyl sulfoxide (DMSO) and formula (1) compound is 6:1, heating, slowly add 1, 2-propylene dichloride and 1, the mixture (1 of 2 dibromo-hexanes, 2-propylene dichloride and 1, the mol ratio of 2-dibromo-hexane is 2:1), temperature rises to 70 DEG C, back flow reaction 7 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to 50-60 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 83%, and purity is 94%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add the mixture (mol ratio of DMF and acetone is 3:1) of DMF and acetone, salt of wormwood and N, mixture (salt of wormwood and the N of N-diisopropylethylamine, the mol ratio of N-diisopropylethylamine is 4:1), the mol ratio of the mixture of described DMF and acetone and formula (2) compound is 9:1, described salt of wormwood and N, the molar weight of the mixture of N-diisopropylethylamine and formula (2) compound is than being 35:1, stir, then the compound and three (3 of formula (3) is added, 6-dioxaheptyl) amine, react 11 hours at temperature is 65 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 60%, and purity is 89%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add the mixture (mol ratio of methyl alcohol and acetone is 1:1) of methyl alcohol and acetone, the mol ratio of the mixture of described methyl alcohol and acetone and formula (4) compound is 6:1, the compound of formula (2) is added after stirring, continue to stir, slowly add sodium bicarbonate, the mol ratio of described sodium bicarbonate and formula (4) compound is 0.1:1, add three (3 again, 6-dioxaheptyl) amine, react 8 hours at 70 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.

Embodiment 9

(1) formula (6) compound is placed in the there-necked flask of dried and clean, add quinoline, the mol ratio of described quinoline and formula (5) compound is 20:1 heating reflux reaction in electricity overlaps, react at temperature is 50 ~ 400 DEG C, monitor with HPLC in reaction process, to raw material primitive reaction completely or product no longer increase, stopped reaction.When temperature is down to below 120 DEG C, add pure frozen water while stirring, be placed in refrigerator and leave standstill crystallization (temperature about 0 DEG C), suction filtration, washing, drying after crystallization, obtain the compound of formula (1); In this step, the yield of gained formula (1) compound is 74%, and purity is 92%;

(2) compound of the formula (1) step (1) prepared adds in clean there-necked flask, add the mixture (mol ratio of methyl alcohol and ethanol is 2:1) of methyl alcohol and ethanol, the mol ratio of the mixture of described methyl alcohol and ethanol and formula (1) compound is 5:1, heating, slowly add 1, 2-ethylene dichloride, temperature rises to 60 DEG C, back flow reaction 8 hours, HPLC monitors reaction, stopped reaction when raw material primitive reaction completely or product no longer increases, treat that temperature is down to less than 40 DEG C, add appropriate 95% ethanol and ethyl acetate carries out crystallization, after crystallization after stirring and crystallizing 1h, carry out suction filtration again, washing, dry, obtain the compound of formula (2), in this step, the yield of gained formula (2) compound is 85%, and purity is 91%,

(3) formula (2) compound that step (2) obtains is added in clean there-necked flask, add quinoline and sodium hydroxide, the mol ratio of described quinoline and formula (2) compound is 6:1, the molar weight of described sodium hydroxide and formula (2) compound is than being 20:1, stir, then the compound of formula (3) and the mixture (mol ratio of tetrabutyl ammonium fluoride and TBAH is 5:1) of tetrabutyl ammonium fluoride and TBAH is added, react 12 hours at temperature is 55 DEG C, suction filtration is carried out after reaction terminates, filter cake washed with dichloromethane is to colourless, filtrate is spin-dried for, add proper amount of acetone, ethyl acetate and sherwood oil crystallization, be precipitated rear suction filtration, filter cake ethyl acetate, petroleum ether, dry, obtain the compound of formula (4), in this step, the yield of gained formula (4) compound is 57%, and purity is 88%,

(4) compound of the formula (4) step (3) obtained adds in clean there-necked flask, add dimethyl sulfoxide (DMSO), the mol ratio of described dimethyl sulfoxide (DMSO) and formula (4) compound is 7:1, the compound of formula (2) is added after stirring, continue to stir, slowly add the mixture (mol ratio of potassium hydroxide and sodium carbonate is 3:1) of potassium hydroxide and sodium carbonate, the mol ratio of the mixture of described potassium hydroxide and sodium carbonate and formula (4) compound is 5:1, add the mixture (mol ratio of Tetrabutyl amonium bromide and tri-n-octyl methyl ammonium chloride is 1:1) of Tetrabutyl amonium bromide and tri-n-octyl methyl ammonium chloride again, react 5 hours at 80 DEG C, monitor with HPLC in reaction process, new peak is had to produce, confirm through mass spectrum, molecular weight is had to be 960, 961, 962 (same isotopic peaks, stack mutually) peak produce, be compound of the present invention, its molecular structural formula is such as formula (5), through extraction, backflow, the methods such as recrystallization are refined further, obtain the compounds of this invention that purity is higher.

The collection of illustrative plates of embodiment 10 compound of the present invention

The compounds of this invention prepared by above-described embodiment 1-9 can obtain, and compound of the present invention is greenish yellow solid.

Can be found out by accompanying drawing 2 ~ 5, compound of the present invention, m.p. (245.7 ~ 248.1 DEG C), ESI-MSm/z:960.1863 [M+Na] ⁺; (mass spectrum demonstrating an isotopic peak)

IR(KBr)ν _max:3022.87，2907.16，1503.24，1395.25，1269.01，1222.65，1099.23，921.807，820.563cm-1

¹HNMR(500MHz,DMSO),δ(ppm):9.86(s,1H,C ₁₄-H),9.50(s,1H,C ₄₀-H),8.88(s,1H,C ₁₁-H),8.84(s,1H,C ₃₇-H),8.21(d,1H,C ₃-H,J＝9Hz),8.14(d,1H,C ₂₉-H,J＝9Hz),8.03(d,1H,C ₅₆-H,J＝9Hz),7.94(d,1H,C ₆₄-H,J＝9Hz),7.80(d,2H,C ₆-H,C ₃₂-H,J＝1.5Hz),7.09(d,1H,C ₅₈-H,J＝7Hz),7.08(d,1H,C ₆₂-H,J＝2.5Hz),7.05(d,1H,C ₁₅-H,J＝8.5Hz),6.99(d,1H,C ₄₁-H,J＝2.0Hz),6.76(m,2H,C ₅₃-H,C ₆₁-H),6.59(s,1H,C ₁₆-H),6.58(s,1H,C ₄₂-H),6.18(d,4H,C ₂₆-H,C ₅₂-H,J＝1.5Hz),5.92(m,1H,C ₆₆-H),5.77(m,1H,C ₆₉-H),5.01(m,4H,C ₆₇-H,C ₇₀-H),4.61(m,4H,C ₂₁-H,C ₂₂-H),4.53(m,4H,C ₄₇-H,C ₄₈-H),4.40(m,2H,C ₉-H/C ₃₅-H),4.07(s,3H,C ₁₉-OCH ₃),3.99(s,3H,C ₄₅-OCH ₃,J＝1.0Hz),3.22(t,2H,C ₆₅-H/C ₆₈-H,J＝13Hz),3.05(m,4H,C ₁₀-H,C ₃₆-H)；

¹³CNMR(500MHz,DMSO),δ(ppm):155.3(C _54,C ₆₀),153.89(C _17,C ₄₃),151.66(C ₁),151.44(C ₂₇).151.25(C ₇),151.18(C ₃₃),149.08(C _2,C ₂₈),146.71(C ₁₈),146.38(C ₄₄),144.11(C ₁ ₄),143.1(C ₄₀),139.46(C ₁₂),139.40(C ₃₈),138.91(C ₆₆),138.63(C ₆₉),134.36(C ₅₇),134.26(C ₆ ₃),132.98(C ₁₁),132.91(C ₃₇),132.06(C ₅₈),131.82(C ₆₄),130.49(C ₅),129.41(C ₃₁),129.24(C ₅₈),128.97(C ₆₂),127.98(C ₅₅),126.01(C ₅₉),122.97(C ₁₅),122.89(C ₄₁),121.64(C ₄),121.60(C ₃₀),116.86(C _13,C ₃₉),116.60(C ₆₇),116.52(C ₇₀),114.21(C _53,C ₆₁),109.81(C ₁₆),109.77(C ₄₂),106.82(C _3,C ₂₉),103.48(C _26,C ₅₂),74.68(C _21,C ₂₂),74.32(C _47,C ₄₈),58.48(C ₉),58.43(C ₃₅),56.82(C ₁₉-OCH ₃),56.54(C ₄₅-OCH ₃),40.02(C ₆₅),39.90(C ₆₈),27.71(C ₁₀),27.66(C ₃₆)。

The antiphlogistic effects test of embodiment 11 the compounds of this invention

1, experiment material, reagent and instrument

Cell strain (Turnover of Mouse Peritoneal Macrophages RAW264.7)

Experiment reagent

Penicillin streptomycin dual anti-(Gibco company); DMEM substratum (Gibco company)

DMSO (Sigma Co., USA's import packing); Foetal calf serum FBS (Gibco company)

MTT (Sigma Co., USA); Lipopolysaccharides LPS (Sigma Co., USA)

Aminoguanidine (Chinese food drug assay institute)

Key instrument

Bechtop (Purifying Equipment Co., Ltd., Suzhou); Home freezer (company of Haier)

CO ₂incubator (ThermoForma company of the U.S.); Inverted microscope (Japanese Nikon company)

Microplate reader (BIO-RAD company);-80 DEG C of refrigerators (Thermo company)

2, the preparation of solution

The preparation of PBS:

NaCl (8.0g); KCl (0.2g); Na ₂hPO ₄12H ₂o (3.5g); KH ₂pO ₂(0.2g) be dissolved in distilled water, adjust ph to 7.4, be settled to 1 liter, 121 DEG C of autoclavings 30 minutes, cool latter 4 DEG C and save backup.The compound method of MTT solution:

Take MTT500mg, be dissolved in the PBS of 100ml, the MTT concentration of configuration is 5mg/ml, with 0.22 μm of membrane filtration to remove the bacterium in solution, puts-20 DEG C and keeps in Dark Place.

The configuration of Griess reagent:

A liquid: take sulfanilamide (SN) 2g, measure strong phosphoric acid 10ml, be dissolved in a certain amount of deionized water, then constant volume is to 200ml

B liquid: take N-1 naphthodiamide hydrochloride 0.2g, be dissolved in a certain amount of deionized water, then constant volume is to 200ml

Cell cultures and digestion: experiment RAW264.7 cell is attached cell, be placed in DMEM nutrient solution (containing 10% foetal calf serum, 0.5% penicillin streptomycin is dual anti-), 5%CO ₂, 37 DEG C of incubators are cultivated.Liquid or digestion is changed according to cell growth state, for subsequent use to exponential phase of growth.

Cell dissociation method: discard original substratum, add 3mlPBS and wash three times, then add 4mlDMEM nutrient solution (containing 10% foetal calf serum, 0.5% penicillin streptomycin is dual anti-), directly blow and beat gently to cell with dropper and all come off in suspension, for subsequent use.

(1) mtt assay measures the compounds of this invention affects RAW264.7 cell-proliferation activity

1, experimental principle

MTT chemistry 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide bromine salt by name, trade(brand)name: tetrazolium bromide.It is a kind of dyestuff of yellow color.It is a kind of method detecting cell survival and growth.Its Cleaning Principle is that the succinodehydrogenase in viable cell plastosome can make exogenous MTT be reduced to water-insoluble bluish voilet Jie Jing formazan and be deposited in cell, and dead cell is without this function.First a ceremonial jade-ladle, used in libation in dimethyl sulfoxide (DMSO) (DMSO) energy dissolved cell, measures its absorbance value with enzyme-linked immunosorbent assay instrument at 570nm wavelength place, can indirectly reflect viable cell quantity.Within the scope of certain cell count, the amount that MTT crystallization is formed is directly proportional to cell count.The method is widely used in cell toxicity test, and its feature is highly sensitive, economical.

2, experimental technique

RAW264.7 cell suspension is inoculated in 96 orifice plates, overnight incubation with 5000/100 μ L.Compound concentration adopts doubling dilution, 6 concentration gradients, the multiple hole of each concentration 3, after every hole adds and cultivates 48 hours with the medicine 100 μ L of substratum dilution, 10%MTT is added in every hole, cultivate 4h at continuing at 37 DEG C, abandoning supernatant, add DMSO150 μ L, the absorption value at 570nm place is read by microplate reader, with blank for 100%, each dosage of computerized compound on the impact of cell viability, by the IC50 value of each compound of spss computed in software.

	Ibuprofen BP/EP	The compounds of this invention	Berberine	Magnolol
					IC ₅₀(μmol/L)	55.67±4.13	85.13±4.62	65.13±5.62	62.36±3.57

Experimental result shows, the compounds of this invention is than the IC of berberine, magnolol and positive control drug Ibuprofen BP/EP ₅₀value is large, and show that the compounds of this invention is compared with berberine, magnolol, its cytotoxicity reduces.

(2) the compounds of this invention induces RAW264.7NO to discharge restraining effect to LPS

Griess method principle: the NO of Hemapoiesis is easily oxidized into NO ₂-, NO ₃-, first by NO ₃-reduction becomes NO ₂-, utilize diazotization reaction and even nitrogenizing reaction to generate color compound, at 540nm wavelength, there is maximum absorption at place.Light absorption value and NO ₂-concentration is proportionate, thus indirectly obtains NO burst size.The change of cell conditioned medium liquid nitrite after the intervention of investigation compound

RAW264.7 cell suspension is with every hole 1 × 10 ⁵individual/100 μ L are inoculated in 96 orifice plates.Blank group (only having RAW264.7 cell) is set, model group (RAW264.7 cell and LPS), with drug treating group (RAW264.7 cell, LPS, medicine), first adopt different pharmaceutical concentration process (100 μMs-3 μMs) process after 2 hours with compound, add LPS again, stimulate after maintaining 24h, draw the nutrient solution 100 μ L in 96 orifice plates, add isopyknic Griess reaction reagent, room temperature reaction 10min, microplate reader 540nm place reads absorbance value.The each dosage of computerized compound in the different treatment time to cell conditioned medium liquid nitrite inhibiting rate.Inhibiting rate (%)=(model group absorbance-each dose drug treatment group absorbance)/(model group absorbance-blank group absorbancy) × 100%.And with the IC that each compound of spss computed in software suppresses NO ₅₀value.Note: all compound DMSO dissolve, and are diluted to required concentration with substratum, the ultimate density of DMSO is 0.1%, adds the DMSO of 0.1% in blank group and model group, and result display does not have toxic action to cell.Aminoguanidine as positive control drug, parallel laboratory test three times.Experimental result GraphPadPrism software analysis.P<0.05, P<0.01 represent that result has significant difference.IC ₅₀calculate by Probit program in SPSS13.0 software.

	Ibuprofen BP/EP	The compounds of this invention	Berberine	Magnolol
					IC ₅₀(μmol/L)	27.25±4.57	28.42±1.62	41.93±5.12	49.64±3.76

Experimental result shows, the compounds of this invention IC ₅₀value is less than berberine Honokiol, and namely the compounds of this invention suppresses NO release action to enhance than berberine Honokiol.

The antibacterial effect test of embodiment 12 the compounds of this invention

(1) experiment material, laboratory apparatus, reagent

1, experiment material

Bacterial strain (clinical separation resistance candida albicans 103,100,953 and J28 are given by Changhai hospital Culture Collection), all experiment bacterial strains all draw plate activation in husky fort glucose agar medium (SDA), after cultivating 2 weeks in 30 DEG C, picking mono-clonal draws plate activation again respectively, get second time gained mono-clonal and put SDA inclined-plane, preserve with for subsequent use in 4 DEG C after cultivating with aforesaid method.

(1) nutrient solution

The preparation of RPMI1640 liquid medium: RPMI1640 (GibcoBRL) 10g, NaHCO ₃2.0g, morpholine propanesulfonic acid (MOPS) (Sigma) 34.5g (0.165M), add tri-distilled water 900ml to dissolve, 1NNaOH adjusts pH to 7.0 (25 DEG C), tri-distilled water is settled to 000ml, 0.22 μm of filtering with microporous membrane is degerming, saves backup after packing in 4 DEG C.

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

Peptone 10g, glucose 40g, agar 18g, add tri-distilled water 900ml and dissolve, and adjustment PH to 7.0, is settled to 1000ml with tri-distilled water, saves backup after autoclaving (121 DEG C, 15min) in 4 DEG C.

(3) YEPD nutrient solution

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

2 reagent

Fluconazole (Fluconazole, FCZ) injection liquid is provided by Dalian pharmaceutcal corporation, Ltd of Pfizer, and berberine hydrochloride (BerberineChloride, BBR) is provided by Shanghai Changhai Hospital.Dimethyl sulfoxide (DMSO) (DimethylSulphoxide, DMSO) Chinese Medicine group Solution on Chemical Reagents in Shanghai Company.

3 laboratory apparatuss

MultiskanMK3 type enzyme mark detector (Finland Labsystems product), water isolation type electro-heating standing-temperature cultivator (Shanghai leap medical apparatus and instruments factory), MJX type intelligence mold incubator (south of the River, Ningbo instrument plant) THZ-82A Desk type constant-temperatureoscillator oscillator (Shanghai leap medical apparatus and instruments factory); SW-CT-IF type superpurgative working table (SuZhou Antai Air Tech Co., Ltd.); Inverted microscope (amershamPharmacia product); Micro sample adding appliance (Finland Finnpette product); 96 porocyte culture plates (Denmark Nunclon Products)

(2) experimental technique

1, the preparation of fungi suspension

Before experiment, a small amount of from the various Candida albicans of picking 4 DEG C of SDA substratum preserved with inoculation circle, be seeded to 1mlYEPD nutrient solution, in 30 DEG C, 200rpm shaking culture, activation 16h makes fungi be in later stage exponential phase of growth.Get this bacterium liquid in 1mlYEPD nutrient solution, after again activating 16h with aforesaid method, with blood cell counting plate counting, with RPMI1640 nutrient solution adjustment bacterial concentration to 1 × 10 ³-5 × 10 ³cFU/ml.

2, the preparation of drug sensitive reaction plate

Get aseptic 96 orifice plates, add RPMI1640 liquid nutrient medium 100 μ l make blank in often arranging No. 1 hole; 3-12 hole respectively adds freshly prepared bacterium liquid 100 μ l; No. 2 holes add bacterium liquid 160 μ l and test-compound solution 40 μ l respectively; No. 12 hole not drug containing, only add bacterium liquid 100 μ l and do Growth positive contrast.Doubling dilution is carried out in 2-11 hole, and make the final drug level in each hole be respectively 64,32,16,8,4,2,1,0.5,0.25 and 0.125 μ g/ml, in each hole, DMSO content is all lower than 1%.All prepare a Quality Control bacterium drug sensitive plate while each preparation drug sensitive plate, each drug sensitive plate is cultivated in 30 DEG C of thermostat containers.

3, the judgement of minimum inhibitory concentration (MIC)

In 30 DEG C of thermostat containers, candidiasis surveys each hole OD value with enzyme micro-plate reader in 620nm after cultivating 24h.The OD value of Positive control wells controls about 0.2, with positive control boring ratio, with the drug level in the minimum concentration hole of OD value decline more than 80% for MIC ₈₀(drug level when fungal growth 80% is suppressed).As the MIC of medicine ₈₀when value exceedes mensuration concentration range, add up by the following method: MIC ₈₀when value is higher than maximum concentration 64 μ g/ml, count " >64 μ g/ml "; MIC ₈₀value for minimum concentration or below minimum concentration time, do not distinguish, all count "≤0.125 μ g/ml ".The equal parallel running of above-mentioned experiment 2 to 3 times, works as MIC ₈₀value is just accepted when accurately can repeat or only differ from a concentration, and using higher concentration as MIC ₈₀value; Work as MIC ₈₀time more than value difference two concentration, then need again to test, until meet the requirements.

	Penicillin	The compounds of this invention	Berberine	Magnolol
					MIC ₈₀(μg/ml)	1.06±0.61	4.68±0.54	3.82±0.32	6.85±0.83

Result shows: the compounds of this invention is to the antibacterial effect of this bacterium not as good as alone berberine, but its successful is better than alone magnolol.

The antitumor anticancer effect test of embodiment 13 the compounds of this invention

For hepatoma H22 cells and malignant myeloid cell lines K562, by inquire into the compounds of this invention to hepatoma H22 cells and malignant myeloid cell lines K562 propagation restraining effect in, study the antitumor antitumous effect that this compound is right.

This experiment adopt SulforhodamineB (SRB) dyeing, trypan blue row dye method, comet electrophoresis technology and the two dye method of AO/EB detect the compounds of this invention to the toxicity of hepatoma H22 cells and malignant myeloid cell lines K562, anticancer proliferation function, to the damage of K562 cell DNA and the apoptotic effect of induction K562 cell.

(1) material, instrument and reagent

1 material and reagent

(DMSO prepares stoste to the compounds of this invention, dilutes before use, and ensureing that DMSO concentration is at least diluted in reaction system is 1 × 10 ^-2), HepG2 cell lines and human leukemia cell line K562 draw from Lanzhou University's Life Science College, substratum RPMI1640 (U.S. Gibcobrl), yellow acyl rhodamine B (SulforhodamineB, SRB, U.S. Sigma), trypan blue (Sigma company), calf serum (Hangzhou folium ilicis chinensis), other chemical reagent is analytical pure.

2 instruments

U.S. PrecisionScientificCO ₂incubator, Bechtop, inverted microscope, opticmicroscope, microplate reader (BIO-RADModel550 and BIO-RADModel680), fluorescent microscope, Horizontal electrophoresis tank, high speed low temperature centrifugal machine, Costar Tissue Culture Plate.

(2) experimental technique

Cell cultures

HepG2 and K562 cell is inoculated in respectively containing in 10% calf serum RPMI1640 substratum, puts 37 DEG C, 5%CO ₂cultivate in incubator, take the logarithm vegetative period cell for experiment.

1, srb assay detects the compounds of this invention to the toxic action of HepG2 cell

Principle SulforhodamineB (SRB) is a kind of protein bound dyestuff, pink, water soluble.SRB can be combined by the basic aminoacids in biomacromolecule, and its OD reading at 511nm and cell count are good linear relationship.Therefore can be used as the quantitative of cell count.

Method: the HepG2 cell of logarithmic phase is with 10 × 10 ⁴ml ^-1concentration be inoculated in 96 well culture plates, after cultivating 24h, in respective aperture, add the compounds of this invention fresh medium 100 μ l of different weaker concn respectively, after continuing cultivation 24,48h, discard nutrient solution, add trichoroacetic acid(TCA) (TCA) the 100 μ l of 10%, after leaving standstill 5min, by orifice plate dislocation 4 DEG C of fixing 1h, outwell stationary liquid, wash 5 times with deionized water, dry.Every hole adds the SRB dyeing 10min of 100 μ l0.4% afterwards, and the acetic acid with 1% washes 5 times, naturally dries.Then every hole adds 150 μ l10mmolL ^-1non-buffered Tris alkali lye (unbufferedTris-basesolution, pH10.5) dissolve and the SRB of protein bound, after making it fully dissolve with vibrator concussion 5min, by the OD value of microplate reader in the every hole of mensuration, 515nm place.Ask its cell proliferation rate (%)=(T-T ₀)/(C-T ₀) × 100.Wherein C represents the cell OD value of control group; T represents the cell OD value of dosing group; T ₀represent the cell OD value during dosing of contrast dull and stereotyped mensuration.At once the cell TCA inoculated in flat board is fixed before dosing.If the final OD value of dosing group is greater than T ₀; Illustrate that cell still grows after dosing.If the final OD value of dosing group is less than T ₀, after dosing is described, cell is killed.Regression equation is utilized to obtain drug level (50%inhibitionconcentration, the IC of 50% increment inhibiting rate ₅₀).From the experimental results, the final cell count of dosing group is less than T ₀, then illustrate that the compounds of this invention cell growth has restraining effect, after dosing, cell is killed.With the IC after the compounds of this invention group and control group process HepG2 cell 24h ₅₀value carrys out the toxic action of more each compound to this cell, IC for example ₅₀be worth as shown in the table:

	Cis-platinum	The compounds of this invention	Berberine	Magnolol
					IC ₅₀(μmol/L)	9.54±0.62	21.08±1.22	22.35±1.42	58.48±4.30

Experimental result shows, the compounds of this invention IC ₅₀value is less than berberine Honokiol, and namely this law people compound strengthens HepG2 cyto-inhibition.

2, trypan blue row dye method detects the compounds of this invention to the toxic action of K562 cell

Principle: viable cell has the ability repelling trypan blue, and due to the complete destruction of film after necrocytosis, namely cell is colored.By in the cell suspension instillation white blood cell count(WBC) plate cell containing trypan blue, observe living cell counting (not being dyed to blue cell) under an optical microscope.

Experimental technique: concentration is 10 × 10 ⁴ml ^-1the K562 cell suspension being in logarithmic phase, with 0.5ml hole ^-1be inoculated in 24 well culture plates.Be placed in 37 DEG C, %CO ₂dosing after preculture 24h in saturated humidity incubator.After continuing cultivation 24,48h, get 0.4% trypan blue solution 0.15ml and add in 0.6ml cell suspension, fully mix, draw a small amount of mixed solution, inject blood cell counting plate cell after dyeing 5min, under opticmicroscope, number viable cell, repeats 3 times, gets its mean number.Ask its cell proliferation rate (%)=(T-T ₀)/(C-T ₀) × 100.Wherein C represents the cell count of control group; T represents the cell count of dosing group; T ₀represent the cell count during dosing of contrast dull and stereotyped mensuration.If the final cell count of dosing group is greater than T ₀, illustrate that cell still grows after dosing.If the final cell count of dosing group is less than T ₀, after dosing is described, cell is killed.Regression equation is utilized to obtain IC ₅₀.From the experimental results, the final cell count of dosing group is less than T ₀, then illustrate that medicine cell growth has restraining effect, after dosing, cell is killed.IC ₅₀be worth as shown in the table:

	Cis-platinum	The compounds of this invention	Berberine	Magnolol
					IC ₅₀(μmol/L)	20.54±2.63	49.58±3.22	53.25±8.44	70.48±6.8

Experimental result shows, the compounds of this invention IC ₅₀value is less than berberine Honokiol, and namely the compounds of this invention strengthens HepG2 cyto-inhibition.

3 the compounds of this invention are to the damage of K562 cell DNA

DNA damage degree determines cell life and death to a certain extent, its detection method SCSE technology.This experiment adopts this technology (concrete grammar detects DNA Damage method with general, does not elaborate) herein,

Detect berberine to the damaging action of K562 cell DNA.By the K562 cell of logarithmic phase with 10 × 10 ⁴ml ^-1concentration be inoculated in 24 well culture plates, be placed in 37 DEG C, 5%CO ₂dosing after preculture 24h in saturated humidity incubator. after continuing to cultivate 48h, collecting cell (1000rmin ^-1centrifugal 5min, removes supernatant).Wash twice with cold PBS, add PBS mixing and make cell suspension.

4 the compounds of this invention induction K562 apoptosis

Principle: acridine orange (AO) through the complete cell of after birth, can embed the green glow that nucleus DNA makes it emitting bright.Ethidium bromide (EB) only can through the impaired cell of after birth, and embedded nuclear DNA, sends out orange fluorescence.At fluorescence microscopy Microscopic observation, visible four kinds of cellular fories: viable cell (V) nuclear chromatin fluoresced green, morphological structure is normal; Viable apoptotic cell (EA) nuclear chromatin jaundice green fluorescence, in pyknosis shape or round bead shape; Non-viable apoptotic cell (LA) nuclear chromatin is orange or orange, and structure is pyknosis shape or round bead shape;

Non-viable non-apoptotic cell (N) nuclear chromatin sends out orange fluorescence, and structure is normal.

Method: the K562 cell in vegetative period of taking the logarithm, with 5 × 10 ⁴ml ^-1be inoculated in 6 orifice plates, every hole 3mL, be placed in 37 DEG C, 5.0%CO ₂saturated humidity incubator in cultivate dosing after 24h.The test drug concentrations of K562 cell is respectively 40,80,160 μm of L ^-1, control group adds isopyknic perfect medium.After continuing cultivation 24,48h, 1000rmin ^-1collecting cell, PBS washes and once gets 100L cell suspension and 4 μ LAO/EB dye liquor (1:1100 μ gmL afterwards ^-1) mixing, smear immediately, excitation wavelength is 570nm Fluirescence observation and counting at least 300 cells.Calculate apoptosis rate (%)=(EA+LA)/(V+N+EA+LA) × 100%.More than test repetition more than 3 times.For the compounds of this invention group of different concns and control group to the more each compound of apoptosis rate after K562 effect 24h to the damaging action of K562 cell DNA.

Experimental result shows, along with the increase of compound concentration, K562 apoptosis rate increases, and illustrates that apoptosis rate and sample concentration are dose-dependence.

The blood sugar reducing function effect test of embodiment 14 the compounds of this invention

Take acarbose as positive control drug, first by vitro enzyme inhibition test, measure the compounds of this invention to the restraining effect of glucuroide, calculate its IC ₅₀value, if this value is less, then can take into account hypoglycemic test in action object.(vitro enzyme inhibition test scheme is similar to above-mentioned anti-inflammatory scheme herein, does not elaborate herein), hypoglycemic test method in body take hyperglycemic rat as study subject, blood sugar reducing function in the body investigating the compounds of this invention.

Hyperglycemic rat modeling and grouping

Choose cleaning grade SD rat (200 scholar 209) 70, male and female half and half, balance raises 3d, detects whole blood blood sugar, without pathoglycemia situation before modeling by blood glucose meter.Rat is divided into control group 10, modeling group 60 at random, sub-cage rearing, experimental session gives basal feed and sterile tap water.Before modeling, water is can't help in 12h fasting, in tail vein injections tetraoxypyrimidine (6omg.kg ^-1body weight), control group injection is with the physiological saline of dosage.Continuous Observation 5 days rear side blood sugar, selects fasting plasma glucose to test use between 13 ~ 20mmolL`.

Choose modeling success diabetes rat 40, be divided into 4 groups at random, often organize 10, set up hyperglycemia model group, Berberine group, the compounds of this invention group and positive control (N1,N1-Dimethylbiguanide) group respectively, remember with 0.5% picric acid solution mark one respectively, by group sub-cage rearing, male and female separately.

Administration: 2% tween 1 solution preparation administrable Berberine, the compounds of this invention and N1,N1-Dimethylbiguanide solution, concentration is 10mg.ml ^-1.Administration group rat presses 100mg.kg ^-1the corresponding liquid 10mL.kg of body weight dose gavage ^-1, body weight, control group and model group rats gavage 2% tween 1 solution 10mL.kg ^-1body weight, every day gastric infusion 1 time, continuous gavage 15 days.Experimentation on animals is carried out in standart animal test room, room temperature 23 scholar 2 DEG C, humidity 50 ~ 60%, and interior lighting and dark alternately 12h/d, change water every day 1 time, within two days, change bedding and padding 1 time, each group freely absorbs drinking-water.

Experimental rat blood sugar detection and function of blood sugar reduction evaluation

After each administration group gastric infusion same day and administration, the 5th, 10,15 day tail vein got blood on an empty stomach, measured each group of rat fasting blood-glucose value by blood glucose meter, respectively organized rat fasting blood-glucose value and blood sugar decline percentage.Blood glucose value × 100% before blood sugar decline percentage=(before experiment the rear blood glucose value of blood glucose value-experiment)/experiment.

Statistical procedures: adopt SPSS13.0 statistical software to carry out statistical procedures to data, all continuous datas represent with " mean+standard deviation (x ± s) ", standard deviation calculates by " n-1 " method, the comparison of many groups sample average, adopt one-way analysis of variance, P<0.05 judges that difference has statistical significance, and P<0.01 judges that difference has significant statistical significance.

By computational analysis, the compounds of this invention is compared with berberine and magnolol, and P<0.05, has statistical significance, variant.Illustrate that the hypoglycemic effect of the compounds of this invention is better than alone berberine and magnolol.

The effect for reducing fat effect test of embodiment 15 the compounds of this invention

With the fat Golden Hamster of high lipid food induction for study subject, after giving the compounds of this invention, detect body weight, liver is heavy, and the indexs such as fat body ratio and blood fat, by observing its fat-reducing effect, and then inquire into its effect for reducing fat.

(1) material

Instrument: supercentrifuge, normal rat test cage, electronic scales

Medicine and reagent: the compounds of this invention, Li Sita difficult to understand, Triglyceride Reagent box, HDL-C test kit, LDL-C test kit.

Animal: male golden yellow gopher 60,

(2) experimental technique

Animal model and grouping: 60 male golden yellow gophers give basal feed and adapt to nursing after 3 days, random selecting 10 is only as Normal group, and remaining its free choice feeding high lipid food that allows (forms: 10% lard, 10% yolk powder, 1% cholesterol, 79% basal feed).After feeding surrounding, fasting can't help water 12h, extracting vein blood after eye socket on an empty stomach, measures its body weight, TC, TG, LDL-C and HDL-C.Successful for modeling Golden Hamster be divided at random 5 groups (n=10): model group, orlistat group, the compounds of this invention is low, in, high dose group, drug intervention 4 weeks, Normal group and model group gavage every day equal-volume physiological saline, Ao Lisi he organize administration every day 42mg/kg, the compounds of this invention is low, in, high dosage is respectively by administration every day 23.35,46.70,70.05mg/kg, Golden Hamster is freely ingested, drinking-water; 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

1 body weight, body length and surplus appetite measure: at duration of test, and each treated animal all takes single cage raise and freely ingest, drinking-water.Within every 3 days, measure a body weight (weighing before gastric infusion) and surplus appetite, and record result.Its body long (nose is to the length of anus), waistline is measured under narcosis, by formula [Lee, s=(body weight) before putting to death ^-(1/3) x10 ³/ body long (cm)] calculate Lee, s index.

2 serum TCs, TG, LDL-C, HDL-C measure: before getting blood, and water 12h is can't help in fasting on an empty stomach, gets blood with glass capillary in retroorbital venous, after fasting 2h, the centrifugal 10min of 5000r/min, gets supernatant liquor,-20 DEG C of preservations, adopt biochemical reagents box to measure TC, TG, LDL-C, HDL-C in serum.

3 livers weights, testis and perirenal fat measure: take the whole fat around liver, perinephric fat and testis and weigh, and calculate the ratio (fat body ratio) of fat/body weight.

(4) data processing

With SPSS17.0 software, statistical study is carried out to the data obtained, data calculated represents with " mean+standard deviation ", carry out one-way analysis of variance, compare between group and adopt t inspection, P<0.05 is that difference has statistical significance, and P<0.01 is that difference has remarkable statistical significance.Percentage hyperlipemia group mouse TC, TG, LDL-C, HDL-C being declined for the compounds of this invention and positive control after medication surrounding tentatively probes into the effect for reducing fat of this medicine.

Result shows: the compounds of this invention is (P<0.05) compared with berberine and magnolol, there is significant difference, and three is compared with positive control drug Li Sita difficult to understand (P<0.01), there is significant significant difference, namely the effect for reducing fat of the compounds of this invention is better than alone berberine or magnolol, but not as good as positive control drug orlistat.

Embodiment 16 the compounds of this invention is tested the action effect of cardiovascular and cerebrovascular diseases

By inquiring into the effect (mainly through observe impact on this mouse right common carotid artery adventitia removal, vascular collagen and elastic plate) of the compounds of this invention on APOE (-/-) mouse early atherosclerosis, study the effect of the compounds of this invention in cardiovascular and cerebrovascular diseases.

(1) materials and methods

1 instrument and equipment

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

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

(3) freezing microtome: German Leica company

(4) Ultralow Temperature Freezer: SANYO GS company

(5) ordinary optical microscope: Japanese Nikon company

(6) polarization microscope: Japanese Olympus company

(7) fluorescent microscope: Japanese Nikon company

(8) ImagePro mono-Plus6.0 image analysis software: MacroMedia company of the U.S.

(9) data gathering and analytical system sPSS17.0: PowerLab company of the U.S.

(10) 7600 automatic analysers: HIT

2 medicines and reagent

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

(2) atorvastatin: Hangzhou Mo Shadong pharmaceutical Co. Ltd

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

(4) talamo: Chemical Reagent Co., Ltd., Sinopharm Group

(5) Hematorylin one Yihong biological stain: Beijing Chemical Plant

(6) phosphate buffered saline buffer (phosphate-bufferedsaline, PBS, 0.01M, pH7.4): take 8gNaCl, 0.2gKCl, 1.44gNa ₂hPO ₄and 0.24gKH ₂pO ₄, be dissolved in 800ml distilled water, by HCI regulator solution pH value to 7.4, last adding distil water is dissolved to IL.

(7) 4% paraformaldehyde liquid: take 409 paraformaldehydes, be placed in flask, add 800ml0.lmol/L phosphoric acid buffer, are heated to 60 DEG C, and magnetic agitation makes powder dissolve completely, finally supplies the PB to 1000ml of 0.lmol/L, fully mixes.

(8) picrosirius staining liquid: 0.1g sirius red, is dissolved in 100ml picric acid saturated solution.

(9) victoria blue staining fluid: victoria blue 2g, dextrin 0.50g, Resorcinol 4g, distilled water 200mL, 5min is boiled in above-mentioned mixing, then 30% ferric chloride solution boiled is added in upper liquid, continue to boil 2min, now constantly stir in colloidal, reduce internal heat after cold filtration, the residue on paper is placed in 60 DEG C of thermostat containers and dries, finally drying residue is dissolved in 75% alcohol of 500ml, add concentrated hydrochloric acid 5ml and phenol 5g again, it is for subsequent use to put room temperature.

(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) laboratory animal and grouping

Take C57BL/6 as male APoE in 6 week age (-/-) mouse 20 of genetic background, be purchased from Department Of Medicine, Peking University.Raise the animal housing of Sino-Japanese Friendship Hospital in meeting secondary animal feeding standard.Indoor purifying air flowing is taken a breath, and keeps room temperature 22 DEG C ~ 25 DEG C, humidity about 50%, 12 hours light and shade cycles (7am-7Pm).Mouse feeder is in standard mouse cage, and mouse cage, drinking bottle, bedding and padding are all through autoclave sterilization.Mouse freely drinks water, and after adaptability feeds 6 days, raises with the high lipid food (60 bore sterilizing radiation treatment) containing lard 21%, cholesterol 1.25% and common mixed feed meal 77.75%.The code of laboratory animal operating process adhere rigidly to Beijing Union Medical College and Ethics Committee of China-Japan Friendship Hospital is carried out, and meets protection of animal ethics to the process of animal.Mouse is divided into 4 groups at random, and the large, medium and small dosage group of blank group, magnolol, often organizes each 5, observes the impact on ApoE (-/-) mouse early atherosclerosis of various dose group after administration.

Medication: according to the people in " pharmacological experimental methodology " (Xu Shuyun edits, the third edition in 2002, People's Health Publisher) and mouse dosage reduction formula, by body weight, Experimental agents dosage is converted.According to the common dose of the clinical recommendation of adult's medication every day, mouse consumption is converted to by adult and the body weight conversion factor 9.01 of mouse, atorvastatin group gives atorvastatin 3mg/kg/d, blank the compounds of this invention group gives physiological saline 0.2ml/kg/d, the compounds of this invention group gives the compounds of this invention heavy dose group 240mg/kg/d, middle dosage group 160mg/kg/d, small dose group 80mg/kg/d with the mode administration of gavage, continues 4 weeks.

The impact of 1 the compounds of this invention on Ap0E (-/-) mouse early atherosclerosis right common carotid artery adventitia removal is drawn materials and section

Draw materials: after experiment mice fasting 12h, with 1% talamo intraperitoneal injection of anesthesia mouse, lain on the back, head and four limbs are fixed on operating table, after alcohol disinfecting, longitudinal incision abdomen chest and skin of neck, successively be separated, cut off tabula, expose heart, after postcava gets blood, poured into by left ventricle with the physiological saline of 0.9%, after blood is rinsed well, expose right common carotid artery sheath, to dissociate right common carotid artery, be about 1cm, rinsed with ice physiological saline, after filter paper blots, being placed on a diameter scribbling OCT frozen section embedding medium is on the small circular fiber board of 3cm, and OC fourth is coated on whole sample, be placed on be frozen in the small beaker filling iso-pentane that existing Liquid nitrogen precooler is about 1min solid-state, be placed in one 80 DEG C of refrigerators and save backup.

(2) cut into slices: with the continuous crosscut of the thickness of 6 μm from below right common carotid artery near-end, leave and take 1 section, be placed on slide glass for every 50 μm, every mouse gets 8 sections.

Pathological examination and analysis: frozen section capable Hematorylin one Yihong (HE) dyes, at the Infiltrating of high power Microscopic observation perivascular inflammatory cell.Range of observation is mouse right common carotid artery and stroma around, because dyeed tangent plane, the length of vessel that appears are different, the area observed is also different, in addition sample inflammatory cell distribution density degree has very big difference, for unified standard, we choose the intensive region of each observed vasculitis cell distribution, often open section total statistical values as this sample of random calculating 8 400 times object lens visual field positive cells on LEICA Image analysis system.

Frozen section HE staining procedure:

1. frozen section, recovers 20min under room temperature

2. cut into slices into HarriS Hematorylin 15min

3. running water 5min

4. 0.5% hydrochloride alcohol differentiation 30sec

5. running water 5min

6. eosin stains 10min

7. running water 5min

8. gradient alcohol dehydration

9. transparent 15minx2 time of dimethylbenzene, middle victory mounting.

Statistical procedures: use SPSSl7.0 software, dosage information adopts mean ± standard deviation (x scholar S) to represent, multiple-group analysis adopts one-way analysis of variance (one-wayANOVA), compare between two between group and adopt LSD, P<0.05 to be that difference has statistical significance.During with heavy dose group, the compounds of this invention group and positive control drug are on the impact of mouse right common carotid artery adventitia removal number of cells, compare the impact of the compounds of this invention on Ap0E (-/-) mouse early atherosclerosis right common carotid artery adventitia removal, and then inquire into its effect in cardiovascular and cerebrovascular diseases.

Group	Atorvastatin	The compounds of this invention	Berberine	Magnolol
					Inflammatory cell number (individual)	5±2	26±3	29±3	38±2

Result shows: the compounds of this invention is (P<0.05) compared with berberine and magnolol, has significant difference, shows that the restraining effect of the compounds of this invention to mouse right common carotid artery adventitia removal is better than berberine or magnolol.

2 the compounds of this invention are on the impact of Ap0E (-/-) mouse early atherosclerosis vascular collagen and elastic plate

Laboratory animal, feeds grouping, draws materials and cuts into slices with testing identical to Ap0E (-/-) mouse early atherosclerosis right common carotid artery adventitia removal.

Pathological examination and analysis

The ruthless scarlet resisdye victoria blue dyeing in frozen section sky, the dyeing of frozen section sirius red chooses section with 0.1% picric acid sirius red dye liquor dyeing, to assess I type and 111 Collagen Type VIs.Every mouse leaves and takes 8 sections, and the collagen content mean value of these 8 sections represents the right common carotid artery collagen content value of every mouse.

Picrosirius staining step:

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

(2) about 5min in dimethylbenzene and straight alcohol (1:1) mixed solution is moved into.

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

(4) washing 3 times is distilled.

(5) with picric acid sirius red dye liquor dye 60min.

(6) raw spirit directly breaks up and dehydration.

(7) dimethylbenzene is transparent, neutral gum mounting.

Frozen section victoria blue dyes: choose section with the dyeing of victoria blue dye liquor, to assess spandex classification and Internal-media thickness.Every mouse leaves and takes 8 sections, and spandex classification and the media thickness mean value thereof of these 8 sections represent the right common carotid artery spandex classification of every mouse and the value of Internal-media thickness.

Victoria blue staining procedure:

20min is recovered under (l) frozen section room temperature.

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

(3) about 30min in victoria blue staining fluid.

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

(5) with distillation washing three times.

(6) dimethylbenzene is transparent, neutral gum sealing.

Statistical procedures: use SPSSl7.0 software, dosage information adopts mean ± standard deviation (x scholar S) to represent, multiple-group analysis adopts one-way analysis of variance (one-wayANOVA), compare between two between group and adopt LSD, P<0.05 to be that difference has statistical significance.During with heavy dose group, the compounds of this invention group and positive controls are to mouse I type, and the impact of the compounds of this invention on Ap0E (-/-) mouse early atherosclerosis vascular collagen and elastic plate is investigated in the impact of III Collagen Type VI content (%) and spandex I grade of sick rate (%).

Group	Atorvastatin	The compounds of this invention	Berberine	Magnolol
					NTx content (%)	21.52±1.63	47.42±1.90	59.42±3.17	62.43±1.51
III Collagen Type VI content (%)	4.53±2.31	41.52±2.50	42.35±1.53	51.48±2.62
					Spandex I grade of disease (%)	60.48±1.52	27.45±2.36	14.30±1.85	13.95±2.74

Result shows: with I type, III Collagen Type VI content (%) and spandex I grade of sick rate (%) are control parameters, the compounds of this invention compares (P<0.05) with berberine and curcumine, there is significant difference, and three compares (P<0.01) with positive controls, there is significant significant difference, and then show that the impact of the compounds of this invention on Ap0E (-/-) mouse early atherosclerosis vascular collagen and elastic plate is better than alone berberine or curcumine, but not as good as positive control drug.

Finally to should be noted that; above embodiment is only in order to illustrate technical scheme of the present invention but not limiting the scope of the invention; although be explained in detail the present invention with reference to preferred embodiment; those of ordinary skill in the art is to be understood that; can modify to technical scheme of the present invention or equivalent replacement, and not depart from essence and the scope of technical solution of the present invention.

Claims

1. a compound, is characterized in that, its structural formula is:

2. a preparation method for compound as claimed in claim 1, is characterized in that, comprise the following steps:

3. the preparation method of compound as claimed in claim 2, it is characterized in that, bromo agent in described step (1) is glycol dibromide, 1,2-ethylidene periodide, 1,2-ethylene dichloride, 1,2-dibromobutane, 1,2-bis-butyl iodide, 1, at least one in 2-propylene dichloride, 1,2-dibromo-hexane.

4. the preparation method of compound as claimed in claim 2, it is characterized in that, alkaline catalysts A in described step (2) and step (3) and alkaline catalysts B is independent separately, be sodium hydroxide, lithium hydroxide, potassium hydroxide, salt of wormwood, sodium carbonate, sodium bicarbonate, saleratus respectively, triethylamine, at least one in DIPEA.

5. the preparation method of compound as claimed in claim 4, it is characterized in that, in described step (2), the molar weight of alkaline catalysts A molar weight and formula (2) compound is than being 0.1:1 ~ 50:1, and in described step (3), the molar weight of alkaline catalysts B and the molar weight of formula (4) compound are than being 0.1:1 ~ 20:1.

6. the preparation method of compound as claimed in claim 2, it is characterized in that, phase-transfer catalyst A in described step (2) and step (3) and phase-transfer catalyst B is independent separately, be at least one in Tetrabutyl amonium bromide, tetrabutyl ammonium fluoride, TBAH, 4-butyl ammonium hydrogen sulfate, three (3,6-dioxaheptyl) amine, benzyl triethyl ammonium bromide, tri-n-octyl methyl ammonium chloride respectively.

7. the preparation method of compound as claimed in claim 2, is characterized in that, in described step (1), the temperature of bromo-reaction is 40 ~ 100 DEG C, and the reaction times is 3 ~ 15 hours; In described step (2), the temperature of nucleophilic substitution reaction is 40 ~ 100 DEG C, and the reaction times is 3 ~ 15 hours; In described step (3), the temperature of secondary nucleophilic substitution reaction is 20 ~ 80 DEG C, and the reaction times is 5 ~ 24 hours.

8. the preparation method of compound as claimed in claim 2, it is characterized in that, in described step (1), the mol ratio of solvent orange 2 A and formula (1) compound is 5 ~ 10:1, and described solvent orange 2 A is at least one in acetonitrile, DMF, methyl alcohol, acetone, ethanol, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), methylene dichloride, dimethyl sulfoxide (DMSO), dioxane; In described step (2), the mol ratio of solvent B and formula (2) compound is 5 ~ 10:1, and described solvent B is at least one in acetonitrile, DMF, ethyl acetate, methyl alcohol, acetone, pyridine, quinoline, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), methylene dichloride, dimethyl sulfoxide (DMSO), dioxane; In described step (3), the mol ratio of solvent C and formula (4) compound is 5 ~ 10:1, and described solvent C is at least one in acetonitrile, DMF, ethyl acetate, methyl alcohol, acetone, pyridine, quinoline, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), methylene dichloride, dimethyl sulfoxide (DMSO), dioxane.

9. the preparation method of compound as claimed in claim 2, it is characterized in that, described step (1) Chinese style (1) compound adopts following methods to be prepared from: reacted at 50 ~ 300 DEG C in solvent D by formula (6) compound, its 9 methyl are sloughed, obtains the compound of formula (1);

10. one kind as claimed in claim 1 compound for the preparation for the treatment of tumour, cancer, hyperglycemia, hyperlipidemia, cardiovascular and cerebrovascular diseases medicine in purposes.