CN102838645B - Polyphenol hydroxy flavone compound with pharmaceutical function and preparation method thereof - Google Patents
Polyphenol hydroxy flavone compound with pharmaceutical function and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the field of medicines and particularly discloses a method for preparing a polyphenol hydroxy flavone compound with a pharmaceutical function. According to the invention, the method comprises the steps of: taking scutellarin and methanol as chlorides of a catalyst, and heating and reacting in a solvent to obtain the polyphenol hydroxy flavone compound. The method is moderate in reacting condition, simple in operation and high in product yield, and is convenient to realize industrialized production; and thus the method has a good industrial application prospect.
Description
Technical field
The invention belongs to field of medicaments, be specifically related to a kind of preparation method with the polyphenol hydroxyl chromocor compound of pharmaceutical use.
Background technology
The aging of people refers to that growth in humans grows after the ripening stage, and with advancing age, a series of degeneration changes occurred in morphological structure and physiological function are processes of human tissue organ's deterioration and decline.Modern medicine is thought, aging is the general performance of the various biochemical reaction of body.In human organism, the moment produces free radical, but has again effective Free-radical ring opening polymerization (as GSH, SOD) simultaneously, and the two is in equilibrium state, thus makes interior free yl maintain normal level.But with advancing age, this balance is destroyed gradually, cause free radical superfluous.Free radical has very strong response capacity, and the many kinds of substance in cell can be made to be oxidized, and infringement microbial film, can also make the macromolecules cross-linking such as protein, nucleic acid, causes apoptosis and physiological function is impaired, decline, thus causes aging.Aging is the inexorable law of vital process, can divide two large classes: one is that physiological is old and feeble, refers to that human body reaches the ripe rear physiology degradation phenomena occurred; Two is pathological seailities, namely on the basis of physiological change, because suffering from some disease or the impact by extraneous factor, and accelerates old and feeble process.
The old and feeble problem of Effect of Anti, not still in order to have good health and a long life, also be improve human survival quality, extend the needs that the mankind effectively " work the age ".China enters aging society, and elderly population increase just at a terrific speed, and astogeny problem is day by day serious, how delaying human body caducity, has become problem in the urgent need to address.
Insomnia, also known as DIMS, clinical manifestation is difficulty falling asleep, early awakening and the length of one's sleep are not enough or poor sleeping quality etc.Insomnia sickness rate increases with the age and increases, and insomnia is very common in the elderly, and according to statistics, China's more than 60 years old prevalence is more than 20% ~ 30%, 70 years old prevalence is 40% ~ 50%, and women is higher than the male sex.Insomnia has a strong impact on the live and work quality of people, causes the decline of function of human body, brings out the generation of various disease.International sleep foundation shows for the enquiry data of the elderly, somnopathy is proportionate with the various diseases with human senescence, can increase the onset risk of the chronic disease relevant to the age such as such as congestive heart failure, renal failure, chronic obstructive pulmonary disease, cerebral apoplexy, Parkinson's disease, diabetes.The hypotype of insomnia has difficulty falling asleep type to have a sleepless night, sleep maintains the insomnia of difficult type, Early morning awakening and psychogenic insomnia, in the elderly sleep maintain difficult type and Early morning awakening comparatively common.According to the course of disease of somnopathy, insomnia can be divided into again acute or short-term insomnia (course of disease <1 month), persistence insomnia (course of disease >4 week) and chronic insomnia (course of disease >6 month).The pathologic process that the treatment of insomnia is had a sleepless night according to patient and determining.Primary insomnia is better to the reaction of pharmacological agent, and insomnia secondary can use pharmacological agent and (or) psychotherapeutics.Treatment should directly for each stage of sleep, and as fallen asleep, sleep maintains, the daytime function of sleep quality or second day.
The medicine of insomnia mainly contains Benzodiazepine (as Decacil and diazepam) and barbiturates (as Sodital, phenylethyl barbituric acid and Amobarbital).Benzodiazepine Department of Pharmacy is by the Benzodiazepine receptor exerts effect of inhibitory neuron one aminobutyric acid (GABA) energy nerve ending, this type of medicine can cause the untoward reactions such as dizzy, sleepy, weak, fine movement is inharmonious when HD, heavy dose of application then can cause ataxia, motor capacity obstacle, and long-time use can cause body resistance and drug dependence.Barbiturate can specific binding in the specific site of GABA acceptor one chloride channel complex surfaces, extend open hour of chloride channel, strengthen the hyperpolarization of postsynaptic membrane, inhibitory neuron discharges, and finally makes central nervous system be suppressed.Such medicine is ascending with dosage, can produce calmness, hypnosis, anticonvulsion and anesthetic action to medication person, to exist after waking up the aftereffect such as dizzy, sleepy, lassitude and significantly Withrawal symptom (as exciting, have a sleepless night, anxiety and convulsions).
In a word, lack one at present and effectively can improve sleep quality, medicine that can delay senility again, that have no side effect, carry out studying and new drug development around this theme, be of great practical significance.
5,6,4 '-trihydroxyflavone-7-O-β-D-Glucose aldehydic acid methyl esters, its structure, such as formula shown in I, is polyphenol hydroxyl saponins chromocor compound, has multiple pharmacological effect, as, existing patent (patent No. 201210001974.2) discloses it in the application improving sleep quality, delay senility.
But current formula I, mainly through plant mode, is extracted and is obtained from the plants such as the root of large-flowered skullcap, rough gentian, Herba Erigerontis, sophora flower.Only extracted from plant by plant mode and obtain a small amount of formula I, rare numbers, be only confined to need for small scale experiments institute, price is extremely expensive, and every gram of value reaches more than six, 7,000 yuan.Because the content of formula I in plant is very low, be only some thousandths of, even ten thousand/a few left and right, extracting and developing is comparatively difficult, and cost is very high, is difficult to mass production, cannot meet current demand.Therefore, how research is by the complete synthesis or semisynthetic mode of chemistry, and the lot-sizeization realizing this compound is produced, and has far-reaching and important meaning to the application and development of this compound.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of artificial large-scale preparation method of formula I, to meet the needs of this compound in medicinal application.
In order to realize foregoing invention object, the invention provides following technical scheme:
A preparation method for formula I, using lamp-dish flower acetic and methyl alcohol and as the muriate of catalyzer reacting by heating and obtaining in specific solvent.
The preparation method of formula I provided by the present invention, directly using commercially available Breviscarpine as reaction raw materials.Before feeding intake, Breviscarpine raw material is without the need to carrying out special purification pre-treatment.The Breviscarpine that preparation method of the present invention adopts, be a kind ofly from the plants such as Herba Erigerontis, extract the Flavonoid substances obtained, its main component is lamp-dish flower acetic.At present, Breviscarpine realizes scale operation at home, and manufacturer reaches family more than 100, and production technology is ripe, and output is large, and the source of goods and steady quality reliably, can arbitrarily have been bought from the market with very low price.In commercially available Breviscarpine, the content of lamp-dish flower acetic is about about 80 ~ 85%.In the embodiment of the present invention, the Breviscarpine raw material used directly is buied from market, and manufacturer is Yuxi, Yunnan Province biological company limited incomparably, and lot identification mark 070821, detect through high performance liquid chromatography (HPLC), lamp-dish flower acetic content is 82.5%.
In the preparation method of formula I provided by the present invention, lamp-dish flower acetic contained in raw material Breviscarpine and methyl alcohol under specific catalyst action, reacting by heating in specific solvent and obtain formula I, reaction principle is shown below:
In the preparation method of formula I provided by the invention, catalyzer is absolutely necessary, and the kind of catalyzer and consumption have vital impact to reaction.Do not add catalyzer in reaction, or the amount of added catalyzer and/or kind are not suitable for, and reaction all may be caused not occur, or other side reaction occurs as oxidation, hydrolysis, alcoholysis etc., cause target product to generate or yield very low.The present inventor, by a large amount of experimental studies, has filtered out the efficiently and directionally catalyzer being applicable to this response characteristic, and has determined the suitable amount scope of catalyzer.
The present inventor, by a large amount of experimental studies, finds that some specific chlorion type compounds have directional catalyzing effect to the present invention's reaction.As preferably, in formula I preparation method, the muriate used as catalyzer is selected from NaCl, KCl, HCl, CaCl
2, MgCl
2, ZnCl
2, FeClx or Fe
2cl
3in one or more mixture.
More preferably, in formula I preparation method used catalyst for being selected from CaCl
2or ZnCl
2in one or more mixture.
In an embodiment of the present invention, contriver is investigated the impact of catalyst levels on formula I productive rate.Experimental result shows, the synthesis of catalyst levels to formula I has material impact.Catalyst levels is too low, does not have katalysis, or catalytic effect is poor, and product yield is low.But after catalyst levels exceedes optimum amount, product yield there will not be obvious rising, too much consumption reagent without practical significance, and can cause the rising of cost.
As preferably, in formula I preparation method, the molar feed ratio of catalyzer and lamp-dish flower acetic is 0.05 ~ 0.5:1.0.
More preferably, in formula I preparation method, the molar feed ratio of catalyzer and lamp-dish flower acetic is 0.1 ~ 0.2:1.0.
In the preparation method of formula I provided by the invention, the selection of solvent system is that can relation react the another key factor that occur smoothly.In an embodiment of the present invention, contriver by lot of experiments, the impact that the selection of comparative study solvent system is synthesized formula I.Result shows, the synthesis of selection to formula I of solvent has material impact.When selecting acetone, ethyl acetate, benzene, toluene, chloroform etc. as solvent, reaction fails normally to occur; And when other condition is identical with operation, when selecting water, acetonitrile, methyl alcohol, dimethyl sulfoxide (DMSO) or DMF etc. for solvent, can there is reaction in various degree in Breviscarpine raw material.In formula I preparation method, highly preferred solvent is one or more the mixture in methyl alcohol or DMF.When being solvent when selecting methyl alcohol, part methyl alcohol wherein participates in reaction as reaction reagent, and unnecessary methyl alcohol then uses as solvent.
In an embodiment of the present invention, contriver is investigated the impact that solvent load synthesizes formula I.Result shows, the synthesis of solvent load to formula I has material impact equally.When solvent load is not enough, lamp-dish flower acetic is difficult to whole dissolving, and cause reaction not thorough, yield is low; On the other hand, if solvent load is too much, reactant concn can be caused too low, be unfavorable for the carrying out reacted equally, yield is lower equally, and can cause the raising of production cost.Experimentally result, the mass ratio of solvent and lamp-dish flower acetic controls at 13.0 ~ 22.0:1.0 comparatively applicable, and best solvent and the mass ratio of lamp-dish flower acetic are 15.0 ~ 20.0:1.0.
The present invention studies by experiment, rationally determines the present invention and reacts suitable temperature of reaction condition and reaction times.
In an embodiment of the present invention, contriver is investigated the impact that temperature of reaction is synthesized formula I.Experimental result shows, the synthesis of temperature of reaction to formula I has material impact.Temperature of reaction is too low, and speed of response is slow, and even do not react, product yield is low; Otherwise if temperature of reaction is too high, side reaction aggravates, and impurity increases, and principal product yield declines, and coloured product deepens.
As preferably, in formula I preparation method, temperature of reaction is 45 ~ 100 DEG C.
More preferably, in formula I preparation method, temperature of reaction used is 50 ~ 80 DEG C.
Except temperature of reaction, there is impact in the performance of reaction times on reaction equally.Generally speaking, temperature of reaction is higher, and the time completed needed for reaction is shorter.In an embodiment of the present invention, contriver, when maintenance temperature of reaction is 50 DEG C and other experiment conditions are constant, has investigated the impact of reaction times on formula I yield.Experimental result shows, under optional test condition, for ensureing that reaction is carried out comparatively thoroughly, the reaction times generally can be controlled in 1 ~ 5 hour, and the preferred reaction times is 1.5 ~ 3.0h.Reaction times is long, and formula I yield is on a declining curve, may be caused by side reaction increases.
Major advantage of the present invention:
The preparation method of formula I provided by the invention, compared with extracting preparation method, has significant advantage with existing plant, is mainly reflected in following several aspect:
1. the large-scale batch production of formula I can be realized.As previously mentioned, current formula I, mainly through plant mode, is extracted and is obtained from the plants such as the root of large-flowered skullcap, Herba Erigerontis, rough gentian.Because the content of this compound in plant is very low, be only some thousandths of to ten thousand/a few left and right, extract comparatively difficulty, output is very low, cannot mass production to meet current demand.The preparation method of formula I provided by the invention, with common commercially available Breviscarpine for raw material, adopts the semi-synthetic mode of artificial chemistry to prepare.Breviscarpine is the Flavonoid substances in oil lamp plant, the content of its main chemical compositions lamp-dish flower acetic in oil lamp plant is very high, general about 1 ~ 10%, be the hundreds of to thousands of times of formula I content, be therefore easy to extensive from plant, low cost and obtain.At present, Breviscarpine realizes large-scale batch production, and domestic production producer is numerous reaches family more than 100, and production technology is ripe, and output is large, and the source of goods and steady quality reliably, can arbitrarily have been bought from the market with very low price.Therefore, by the preparation method of formula I provided by the invention, the large-scale batch production of formula I can be realized.
2, production cost is low.As previously mentioned, the content of formula I in plant is very low, and adopt the preparation of plant extracting mode, yield is low, needs the plant resources of at substantial preciousness.In addition, plant is extracted preparation method and is implemented comparatively difficulty, need to consume a large amount of Extraction solvent, crude extract also needs comprehensively to adopt the methods such as polyamide column chromatography, silica gel column chromatography and recrystallization repeatedly isolation andpurification repeatedly, just can reach medicinal required purity, therefore, production cost is very expensive, and every gram of cost reaches more than more than 6000 yuan.Comparatively speaking, the source chemicals that formula I preparation method adopts is cheap to be easy to get, and technique is brief, and yield is high, and production cost is low, production formula I per kilogram cost about about 2000 yuan, about being only three thousandths of plant extraction preparation method.Therefore, formula I preparation method of the present invention has white war advantage.
3. reaction conditions is gentle, and simple to operate, product yield is high.The maximum innovation point of the present invention is to adopt specific muriate as efficiently and directionally catalyzer, directly uses Breviscarpine to be raw material, under lower temperature condition, has synthesized formula I with high yield.Because the present invention have selected suitable reaction solvent and system, Breviscarpine raw material in preparation method of the present invention need not refinement treatment in advance, wherein contained impurity component can not disturb main reaction, successfully realizes being separated with principal product again after reaction terminates by simple aftertreatment.The preferred catalyzer of preparation method of the present invention is CaCl
2or ZnCl
2, reaction solvent is methyl alcohol or DMF, temperature of reaction is 50 ~ 80 DEG C, reaction times 1.5 ~ 3.0h, with this understanding, formula I can reach 95 ~ 98%, have catalyzer efficiently and directionally strong, cheap and easy to get, reaction conditions is gentle, and the time is short, simple to operate, the distinguishing features such as product yield is high, are therefore convenient to industrialization and produce, have very strong practicality.
Formula I prepared by the present invention, pharmaceutically tool has been widely used.Described purposes comprises and is used for the treatment of insomnia, improves sleep quality, extends the deep sleep time, Improving memory power, the diseases such as treatment and prevention memory loss, hypomnesis, senile dementia.Described purposes also comprises for delaying human body caducity, the clinical application of the aspects such as prolongs life.
Formula I prepared by the present invention can form pharmaceutical preparation with pharmaceutically acceptable auxiliary material, is applied to clinical.Described pharmaceutically acceptable auxiliary material, comprises all kinds of SOLVENTS, thickening material, solubilizing agent, solubility promoter, inclusion agents, filler, sanitas, stablizer, correctives, tinting material and sweetener etc.Specifically, described pharmaceutically acceptable auxiliary material, including, but not limited to starch, cyclodextrin, beta-cyclodextrin, Yelkin TTS, fatty oil, soybean oil, peanut oil, sesame oil, water, alcohol, glucose, N.F,USP MANNITOL, tween, glycerine, sodium-chlor, magnesium sulfate etc.
Formula I prepared by the present invention is applied to clinical with various pharmaceutical preparation.
Formula I prepared by the present invention, can be made into oral preparations or injection formulations.Described oral preparations be specially tablet, capsule, mixture, oral liquid, containing agent and dripping pill etc.Described injection formulations is specially injection liquid, infusion solutions or lyophilized injectable powder.
Formula I prepared by the present invention, also can be made into external preparation and use, described external preparation comprises liniment, patch, cataplasma, sprays, aerosol etc.
Formula I prepared by the present invention, also can be made into various Novel medicine feeding formulation application in clinical, and described Novel medicine feeding preparation can be various inclusion preparation, targeting preparation, at regular time and quantity drug-delivery preparation etc.Described inclusion preparation comprises beta-cyclodextrin inclusion preparation, starch inclusion preparation, protein inclusion preparation, high-fat emulsion inclusion preparation, chitosan inclusion preparation etc.
Formula I prepared by the present invention can also form pharmaceutical composition with other medicines or excipient substance, and is applied to clinical.Described pharmaceutical composition is made up of two or more material comprising formula I.
Embodiment
The invention discloses a kind of compound and preparation method thereof, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply the technology of the present invention.
In the embodiment of described formula I synthesis below, Breviscarpine raw material used, by Kunming, drug research institute of pharmacy group directly buys from market, manufacturer is Yuxi, Yunnan Province biological company limited incomparably, lot identification mark 070821, detect through high performance liquid chromatography (HPLC), lamp-dish flower acetic content is 82.5%.Chemical reagent methyl alcohol used, acetonitrile, dimethyl sulfoxide (DMSO), dimethyl sulfoxide (DMSO), DMF, acetone, ethyl acetate, benzene, toluene, chloroform, sherwood oil, NaCl, KCl, HCl, CaCl
2, MgCl
2, ZnCl
2, FeCl
2and Fe
2cl
3be chemical pure, purchased from Kunming Shan Dian Chemical Co., Ltd., water is purified water.
Following examples 1 ~ 9, when keeping other condition and operating method is constant, the impact that the selection that compared for different catalysts is synthesized formula I, it the results are shown in Table 1.In each embodiment reaction product formula I all through HPLC,
1hNMR,
13cNMR, IR and UV qualification and inspection.
(used catalyst is CaCl in the synthesis of embodiment 1 formula I
2)
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.Reaction solution adopts vacuum concentration to reclaim methyl alcohol, and when solution to be concentrated volume is reduced to about 1/3 of original volume, stop concentrated, concentrated solution leaves standstill and is cooled to room temperature, has a large amount of product crystallization.Vacuum filtration, divide to take out for three times with 30mL cold methanol and wash, drain, products therefrom proceeds in 250mL ethyl acetate-light petrol (3:1), is heated to the 30min that boils as far as possible, sets to 0-5 DEG C of refrigerator and cooled but more than 4h.Take out crystallized product, vacuum filtration, takes out with 30mL cold ethyl acetate and washes three times, finally taking out with about 15mL sherwood oil washes once, drains as far as possible, puts dry more than 48h in 60 DEG C of vacuum drying ovens, obtain the formula I 22.8g of light yellow loose powders, yield 95.8%.
(used catalyst is ZnCl in the synthesis of embodiment 2 formula I
2)
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.68g ZnCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 23.3g of light yellow loose powders, yield 97.9%.
(used catalyst is MgCl in the synthesis of embodiment 3 formula I
2)
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.48g MgCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain light yellow loose powders formula I 19.3g, yield 81.1%.
(used catalyst is FeCl in the synthesis of embodiment 4 formula I
2)
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.63g FeCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain light yellow loose powders formula I 17.5g, yield 73.5%.
(used catalyst is FeCl in the synthesis of embodiment 5 formula I
3)
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.81g FeCl
3(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain light yellow loose powders formula I 18.7g, yield 78.6%.
The synthesis (used catalyst is HCl) of embodiment 6 formula I
Get Breviscarpine raw material 28.0g and (amount to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add the concentrated hydrochloric acid (containing HCl0.005mol) that methyl alcohol 500g, 0.50g concentration is 36%, in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain light yellow loose powders formula I 13.2g, yield 55.5%.
The synthesis (used catalyst is NaCl) of embodiment 7 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.29g NaCl (0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain light yellow loose powders formula I 19.8g, yield 83.2%.
The synthesis (used catalyst is KCl) of embodiment 8 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.37g KCl (0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain light yellow loose powders formula I 16.2g, yield 68.1%.
The synthesis (not using catalyzer) of embodiment 9 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, in 50 DEG C of heated and stirred reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.The impact that the selection of table 1 different catalysts is synthesized formula I
Above-mentioned experiment shows, compound has obvious catalysed promoted effect to this reaction, and the synthesis of the selection of different catalysts to formula I has material impact.Muriatic catalysis action order is ZnCl
2> CaCl
2> NaCl > MgCl
2> FeCl
3> FeCl
2> KCl > HCl, wherein, especially CaCl
2and ZnCl
2katalysis best.
Following examples 10 ~ 14 and embodiment 1 and embodiment 9, with CaCl
2for catalyzer, when keeping other condition and operating method is constant, compared for the impact that catalyst levels synthesizes formula I, it the results are shown in Table 2.In each embodiment reaction product all through HPLC,
1hNMR,
13cNMR, IR and UV qualification and inspection.
The synthesis (catalyzer and lamp-dish flower acetic mol ratio are 0.05:1.0) of embodiment 10 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.28g CaCl
2(0.0025mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 9.9g of light yellow loose powders, yield 41.6%.
The synthesis (catalyzer and lamp-dish flower acetic mol ratio are 0.08:1.0) of embodiment 11 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.44g CaCl
2(0.0040mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 19.4g of light yellow loose powders, yield 81.5%.
The synthesis (catalyzer and lamp-dish flower acetic mol ratio are 0.15:1.0) of embodiment 12 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 0.83g CaCl
2(0.0075mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 22.9g of light yellow loose powders, yield 96.2%.
The synthesis (catalyzer and lamp-dish flower acetic mol ratio are 0.20:1.0) of embodiment 13 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 1.11g CaCl
2(0.01mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 22.7g of light yellow loose powders, yield 95.4%.
The synthesis (catalyzer and lamp-dish flower acetic mol ratio are 0.50:1.0) of embodiment 14 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 500g, 2.78g CaCl
2(0.025mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 23.0g of light yellow loose powders, yield 96.6%.
The impact that table 2 catalyst levels synthesizes formula I
As shown in Table 2, the synthesis of catalyst levels to formula I has material impact.Catalyst levels is too low, does not reach katalysis, or catalytic effect is poor, and product yield is lower.But after catalyst levels exceedes optimal economic consumption, product yield can not occur obviously rising with the increase of catalyst levels, consumes reagent too much without practical significance, and can cause the rising of cost.Consider, determine in this building-up reactions, the optimum mole ratio scope of catalyzer and lamp-dish flower acetic is 0.1 ~ 0.2:1.0.
Following examples 15 ~ 23 and embodiment 1, when keeping other condition and operating method is constant, the impact that the selection having investigated solvent is synthesized formula I, the results are shown in Table 3.In each embodiment reaction product all through HPLC,
1hNMR,
13cNMR, IR and UV Identification and detection.
The synthesis (solvent is water) of embodiment 15 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, water 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 18.1g of light yellow loose powders, yield 76.1%.
The synthesis (solvent is acetonitrile) of embodiment 16 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, water-acetonitrile 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 12.7g of light yellow loose powders, yield 53.4%.
The synthesis (solvent is dimethyl sulfoxide (DMSO)) of embodiment 17 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, dimethyl sulfoxide (DMSO) 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 19.3g of light yellow loose powders, yield 81.1%.
The synthesis (solvent is DMF) of embodiment 18 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 23.2g of light yellow loose powders, yield 97.5%.
The synthesis (solvent is acetone) of embodiment 19 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, acetone 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.
The synthesis (solvent is ethyl acetate) of embodiment 20 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, ethyl acetate 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.
The synthesis (solvent is benzene) of embodiment 21 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, benzene 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.
The synthesis (solvent is toluene) of embodiment 22 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, toluene 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.
The synthesis (solvent is chloroform) of embodiment 23 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, chloroform 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.
The impact that table 3 different solvents synthesizes formula I
From the above, the synthesis of selection to formula I of solvent has material impact.When selecting acetone, ethyl acetate, benzene, toluene, chloroform etc. as reaction solvent, Breviscarpine and methyl alcohol do not react production I under chloride catalyst effect; And when other condition is identical with operation, when selecting water, acetonitrile, methyl alcohol, dimethyl sulfoxide (DMSO) or DMF to be reaction solvent, can there is different reactions in Breviscarpine raw material, obtains the formula I of different productive rate.From experimental result, this reaction is to select methyl alcohol or DMF for the yield obtained during solvent is for height.When being solvent when selecting methyl alcohol, part methyl alcohol wherein participates in reaction as reaction reagent, and unnecessary methyl alcohol then uses as solvent.
Following examples 24 ~ 30 and embodiment 18 take DMF as solvent, and when keeping other condition and operating method is constant, the impact that the consumption having investigated solvent synthesizes formula I, the results are shown in Table 4.In each embodiment reaction product all through HPLC,
1hNMR,
13cNMR, IR and UV Identification and detection.
The synthesis (mass ratio of solvent and lamp-dish flower acetic is 4.33:1.0) of embodiment 24 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 100g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 4.7g of light yellow loose powders, yield 19.7%.
The synthesis (mass ratio of solvent and lamp-dish flower acetic is 8.66:1.0) of embodiment 25 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 200g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 11.8g of light yellow loose powders, yield 49.6%.
The synthesis (mass ratio of solvent and lamp-dish flower acetic is 12.99:1.0) of embodiment 26 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 300g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 17.8g of light yellow loose powders, yield 74.8%.
The synthesis (mass ratio of solvent and lamp-dish flower acetic is 15.15:1.0) of embodiment 27 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 350g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 22.1g of light yellow loose powders, yield 92.9%.
The synthesis (mass ratio of solvent and lamp-dish flower acetic is 19.48:1.0) of embodiment 28 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 450g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 23.3g of light yellow loose powders, yield 97.9%.
The synthesis (mass ratio of solvent and lamp-dish flower acetic is 21.65:1.0) of embodiment 29 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 500g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 21.3g of light yellow loose powders, yield 89.5%.
The synthesis (mass ratio of solvent and lamp-dish flower acetic is 25.97:1.0) of embodiment 30 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 600g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 2h.All the other operate with embodiment 1, obtain the formula I 15.9g of light yellow loose powders, yield 66.8%.
The impact that table 4 solvent load synthesizes formula I
From the above, the synthesis of solvent load to formula I has material impact equally.When solvent load is not enough, lamp-dish flower acetic is difficult to whole dissolving, and cause reaction not thorough, yield is low; On the other hand, if solvent load is too much, reactant concn can be caused too low, be unfavorable for the carrying out reacted equally, yield is lower equally, and can cause the raising of production cost.According to above-mentioned experimental result, the mass ratio of solvent and lamp-dish flower acetic controls at 13.0 ~ 22.0:1.0 comparatively applicable, and best solvent and the mass ratio of lamp-dish flower acetic are 15.0 ~ 20.0:1.0.
Following examples 31 ~ 38 and embodiment 18, when keeping other condition and operating method is constant, having investigated the impact that temperature of reaction is synthesized formula I, having the results are shown in Table 5.In each embodiment reaction product all through HPLC,
1hNMR,
13cNMR, IR and UV Identification and detection.
The synthesis (temperature of reaction 20 DEG C) of embodiment 31 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 20 DEG C of stirring reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.
The synthesis (temperature of reaction 30 DEG C) of embodiment 32 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 30 DEG C of stirring reaction 2h.Reactant is through TLC and HPLC(high performance liquid chromatography) detect, prove that Breviscarpine raw material does not react production I.
The synthesis (temperature of reaction 40 DEG C) of embodiment 33 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 40 DEG C of stirring reaction 2h.All the other operate with embodiment 1, obtain the formula I 8.9g of light yellow loose powders, yield 37.4%.
The synthesis (temperature of reaction 45 DEG C) of embodiment 34 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 45 DEG C of stirring reaction 2h.All the other operate with embodiment 1, obtain the formula I 18.8g of light yellow loose powders, yield 79.0%.
The synthesis (temperature of reaction 60 DEG C) of embodiment 35 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 60 DEG C of stirring reaction 2h.All the other operate with embodiment 1, obtain the formula I 23.3g of light yellow loose powders, yield 97.9%.
The synthesis (temperature of reaction 80 DEG C) of embodiment 36 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 80 DEG C of stirring reaction 2h.All the other operate with embodiment 1, obtain the formula I 23.1g of light yellow loose powders, yield 97.1%.
The synthesis (temperature of reaction 100 DEG C) of embodiment 37 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 100 DEG C of stirring reaction 2h.All the other operate with embodiment 1, obtain the formula I 20.0g of light yellow loose powders, yield 84.0%.
The synthesis (temperature of reaction 120 DEG C) of embodiment 38 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 120 DEG C of stirring reaction 2h.All the other operate with embodiment 1, obtain the formula I 15.6g of light yellow loose powders, yield 65.5%.
The impact that table 4 temperature of reaction is synthesized formula I
From the above, the synthesis of temperature of reaction to formula I has material impact.Temperature of reaction is too low, and speed of response is slow, and even do not react, product yield is low; Otherwise if temperature of reaction is too high, side reaction aggravates, and impurity increases, and principal product yield declines, and coloured product deepens.According to above-mentioned experimental result, temperature of reaction controls at 45 ~ 100 DEG C comparatively applicable, and preferred range of reaction temperature is 50-80 DEG C.
Following examples 39 ~ 44 and embodiment 18, when keeping other condition and operating method is constant, having investigated the impact that the reaction times synthesizes formula I, having the results are shown in Table 5.In each embodiment reaction product all through HPLC,
1hNMR,
13cNMR, IR and UV Identification and detection.
The synthesis (reaction times 0.5h) of embodiment 39 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 0.5h hour.All the other operate with embodiment 1, obtain the formula I 11.1g of light yellow loose powders, yield 46.6%.
The synthesis (reaction times 1.0h) of embodiment 40 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 1.0h hour.All the other operate with embodiment 1, obtain the formula I 18.7g of light yellow loose powders, yield 78.6%.
The synthesis (reaction times 1.5h) of embodiment 41 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 1.5h hour.All the other operate with embodiment 1, obtain the formula I 22.9g of light yellow loose powders, yield 96.2%.
The synthesis (reaction times 3.0h) of embodiment 42 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 3.0h hour.All the other operate with embodiment 1, obtain the formula I 23.1g of light yellow loose powders, yield 97.1%.
The synthesis (reaction times 5.0h) of embodiment 43 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 5.0h hour.All the other operate with embodiment 1, obtain the formula I 21.7g of light yellow loose powders, yield 91.2%.
The synthesis (reaction times 10.0h) of embodiment 44 formula I
Get Breviscarpine raw material 28.0g (amounting to lamp-dish flower acetic 23.1g, 0.05mol), put in 1000mL round-bottomed flask, add methyl alcohol 100g, DMF 400g, 0.56g CaCl
2(0.005mol), in 50 DEG C of heated and stirred reaction 10.0h hour.All the other operate with embodiment 1, obtain the formula I 19.3g of light yellow loose powders, yield 81.1%.
The impact that table 6 reaction times synthesizes formula I
From the above, the synthesis of reaction times to formula I has certain influence.Under optional test condition, for ensureing that reaction is carried out comparatively thoroughly, the reaction times generally can be controlled in 1 ~ 5 hour, comparatively 1.5 ~ 3.0h of preferred reaction times.Reaction times is long, and formula I yield is on a declining curve, may be caused by side reaction increases.
The experimental study of embodiment 45 formula I improving water flood
1. reagent, reagent and material
Formula I, drug research institute of Kunming Medicine Group Stock Co., Ltd provides.Vetanarcol, the gloomy Bioisystech Co., Ltd in Wal, Xi'an provides.
Kunming mouse, body weight is 18 ~ 22g, 180, male and female half and half, and Kunming Medicine Group Stock Co., Ltd's Animal Lab. provides, production licence number SCXK (Yunnan) 2005-0006, occupancy permit SYXK (Yunnan) 2005-0006.
2. test method
2.1 animal grouping and administrations
Laboratory animal is divided into 3 batches, be designated as that experiment I criticizes (on the impact of Mouse Weight, directly sleep experiments, extending the vetanarcol experiment length of one's sleep), experiment II criticizes (vetanarcol sub-threshold dose hypnosis experiment), experiment III criticizes (experiment of vetanarcol Sleep latency) respectively, often criticize animal and be divided into 4 groups at random according to body weight. often group 15.If 5,6, the basic, normal, high dosage group of 4 '-trihydroxyflavone-7-O-β-D-Glucose aldehydic acid methyl esters (be respectively 25,50,150mg/kg body weight dose group), gastric infusion, control group gavage gives equivalent distilled water.Every day gavage 1 time, continuous 30 days.
2.2 direct sleep laboratory
Observe the given the test agent that tested treated animal gives 3 dosage, after control group gives isopyknic distilled water, whether occur phenomenon of sleeping.Sleep is index with righting reflex loss.When mouse is placed in supine position, body position can be righted immediately, as can not the person of righting more than 30 ~ 60 seconds, namely think and righting reflex loss enter sleep.Righting reflex recovers to be animal awakening, and righting reflex loss is the animal sleep time to recovering during this period of time.Record control group and tested group of sleep number of animals and the length of one's sleep
2.3 lengths of one's sleep extending vetanarcol induction tested
First preliminary experiment is carried out before doing formal experiment. determine to make animal 100% fall asleep but the vetanarcol dosage (40mg/kg body weight) that the length of one's sleep can not be made long, formally test with this dosage.After animal last gives tested material, occur 15min before peak effect, to each treated animal abdominal injection vetanarcol, injection volume is 40mg/kg body weight, with mouse righting reflex loss for sleep index.Observe tested material to the prolongation effect of the length of one's sleep that vetanarcol are induced.
2.4 vetanarcol sub-threshold dose hypnosis test
First preliminary experiment is carried out before doing formal experiment, determine pentobarbital sodium sub-threshold lull dosage, namely the maximum sub-threshold dose of vetanarcol (vetanarcol 25mg/kg body weight) that do not disappear of 80% ~ 90% mouse righting reflex, formally tests with this dosage.After animal last gives tested material, there is 15min before peak effect, to each treated animal abdominal injection vetanarcol, reach more than 1min as sleep judging criterion using mouse righting reflex loss.Observe the number of animals giving each treated animal in vetanarcol 30min and occur to sleep.
2.5 vetanarcol inducing mouse Sleep Latency Test
First carry out preliminary experiment before doing formal experiment, determine animal 100% is fallen asleep but the vetanarcol dosage (300mg/kg body weight) not making the length of one's sleep long, formally test with this dosage.Animal last gives 15min after tested material, and to each treated animal abdominal injection vetanarcol, injection volume is 300mg/kg body weight, take righting reflex loss as index, observes tested material to the impact of vetanarcol inducing mouse Sleep latency.
3. experimental result
3.1 formula I are on the impact of the weight of animals
The each dosage group of formula I compares with control group.Experimental session body weight there are no significant difference (P>0.05), points out this test conditions compounds of Formula I to have no significant effect Mouse Weight, in Table l.
Table 1 tested material is on the impact of experiment mice body weight
3.2 formula I are to the direct sleep effect of animal
Each dosage group and control group. after gastric infusion all there is not phenomenon of sleeping in mouse, and under pointing out this experiment condition, formula I acts on without direct sleep experiment mice.In table 2.
Table 2 tested material is to the direct sleep effect of experiment mice
3.3 formula I are on the impact of the mouse sleep time that vetanarcol are induced
Table 3 tested material is on the impact of the mouse sleep time that vetanarcol are induced
From table 3, after giving the test medicine of mouse various dose, compare with control group, basic, normal, high dosage group extends 16.2% (P ﹥ 0.05), 65.4%(P ﹤ 0.05 respectively to the mouse sleep time that vetanarcol are induced) and 111.4% (P ﹤ 0.01), show that formula I has significant prolongation effect to the mouse sleep time that vetanarcol are induced, and in certain dose-dependence.
3.4 test medicine are on the impact of the mice sleep incidence that vetanarcol sub-threshold dose is induced
Table 4 tested material is on the impact of the mice sleep incidence that vetanarcol sub-threshold dose is induced
From table 4, after giving the test medicine of mouse various dose, compare with control group, basic, normal, high dosage group improves 50.4%(P ﹤ 0.05 respectively to the mice sleep incidence that vetanarcol are induced), 150.3% (﹤ 0.01), 351.1% (﹤ 0.01), show the mice sleep incidence effect of increasing significantly that test medicine is induced vetanarcol, and in certain dose-dependence.
The preclinical impact of mice sleep that 3.5 test medicine are induced vetanarcol
The preclinical impact of mice sleep that table 5 tested material is induced vetanarcol
From table 5, after giving the test medicine of mouse various dose, compare with control group, the mice sleep that basic, normal, high dosage group is induced vetanarcol shortens 1.2%(P ﹥ 0.05 latent period respectively), 11.1% (﹥ 0.05), 28.1% (P ﹤ 0.01), show that test medicine has certain shortening effect to the mice sleep that vetanarcol are induced latent period, and in certain dose-dependence.
Embodiment 46 formula I is tested the influence of life span of drosophila melanogaster
1. reagent, reagent and material
Formula I, drug research institute of Kunming Medicine Group Stock Co., Ltd provides.
U.S.'s wild-type drosophila melanogaster, feeding environment: temperature 25 ± 1 DEG C, relative humidity 60 ± 5%, 12h illumination, the dark alternate culture of 12h, Kunming Institute of Botany provides.
2. test method
Collect the new fruit bat adult sprouted wings in 24h, through etherization, distinguish male and female individuality, after weighing respectively, be divided into 5 groups: 1 control group and 4 experimental group containing different concns formula I at random.Often organize fruit bat 100, male and female half and half, put in the test tube of 3cm × 10cm that substratum is housed respectively, often pipe 25.Control group gives conventional corn powder substratum, experimental group gives containing 0.005% respectively, 0.025%, 0.075% and 0.550% 5, the maize powder medium of 6,4 '-trihydroxyflavone-7-O-β-D-Glucose aldehydic acid methacrylate powder, test tube stands in incubator and cultivates, experimental temperature is 25 ± 1 DEG C, relative humidity 60 ± 5%.Within every 4 days, change 1 freshly prepared substratum, every day observed and recorded drosophila survival number and death toll, until the whole death of fruit bat.Calculate the dead number of days of half, mean lifetime and Mean longest life.
3. experimental result
Table 6 formula I is on the impact of life span of drosophila melanogaster
From table 6, compared with control group, there is certain prolongation the half death time of each concentration group to fruit bat.Compared with male control group, reagent substrate concentration is the half death time of 0.005%, 0.025%, 0.075%, 0.550% male group extend 1.4%, 8.7%, 16.6%, 14.1% respectively; Compared with female control group, reagent substrate concentration is the half death time of 0.005%, 0.025%, 0.075%, 0.550% female group extend 2.5%, 10.5%, 15.8%, 16.4% respectively.
Table 6 also shows, the mean lifetime of 0.025%, 0.075%, 0.550% reagent substrate concentration equal energy significant prolongation fruit bat and Mean longest life.Compared with male control group, reagent substrate concentration is that the mean lifetime of 0.005%, 0.025%, 0.075%, 0.550% male group extends 1.4%, 9.0%, 16.7%, 13.3% respectively; Compared with female control group, concentration is that the mean lifetime of 0.005%, 0.025%, 0.075%, 0.550% female group extends 1.6%, 11.0%, 15.1%, 16.4% respectively.Compared with male control group, reagent substrate concentration is that the maximum life span of 0.005%, 0.025%, 0.075%, 0.550% male group extends 0.4%, 5.4%, 11.7%, 12.5% respectively; Compared with female control group, reagent substrate concentration is that the maximum life span of 0.005%, 0.025%, 0.075%, 0.550% female group extends 1.4%, 7.1%, 10.8%, 10.1% respectively.As can be seen here, 5,6,4 '-trihydroxyflavone-7-O-β-D-Glucose aldehydic acid methyl esters effectively can extend half death time, mean lifetime and the highest mean lifetime of fruit bat.
The mechanism experiment that embodiment 47 formula I delays senility
1. reagent, reagent and material
Formula I, drug research institute of Kunming Medicine Group Stock Co., Ltd provides.
Mda (MDA) measures test kit, and lot number is 20080925; Superoxide-dismutase (SOD) measures test kit, and when specifying that SOD inhibiting rate in every milliliter of reaction solution reaches 50%, corresponding SOD amount is 1 nitrite unit (NU), and lot number is 20080629; Selenoperoxidase (GSH-PX) measures test kit, and specify that every 4 μ l whole bloods are at 37 DEG C of reaction 5min, deduct the effect of non-enzymatic reaction, making GSH concentration in reaction system reduce by 1 μm of ol/L is 1 enzyme activity unit, and lot number is 20080918; Active oxygen (ROS) measures test kit, specifies that every milliliter of serum reacts 1min at 37 DEG C, makes H in reaction system
2o
2it is 1 active oxygen unit that concentration reduces 1mmol/L, and lot number is 20080721.Above test kit builds up Bioengineering Research Institute by Nanjing and provides.
Kunming mouse, at 15 monthly ages, body weight 50 ± 2g, Kunming Medicine Group Stock Co., Ltd's Animal Lab. provides, production licence number SCXK (Yunnan) 2005-0006, occupancy permit SYXK (Yunnan) 2005-0006.
2. test method
Aged kunming mice 40, male and female half and half, are divided into 4 groups: Normal group at random, low dose group (50mg/kg), middle dosage group (100mg/kg) and high dose group (200mg/kg).The test medicine formula I gavage of each dosage group mouse prescribed dose, the distilled water gavage of Normal group mouse same volume.Every day 1 time, continuous 30 days.In addition, within every 10 days, weigh 1 time.24h after last gavage, plucks eyeball blood sampling.According to test kit operation instructions, measure mouse red blood cell SOD respectively active, serum SOD-PX activity and red corpuscle MDA content, serum ROS content.
3. experimental result
Table 7 formula I delays senility mechanism
Above-mentioned table 7 result shows, it is active that formula I can significantly improve SOD and GSH-PX in aged mouse body, reduces ROS content and MDA content in aged mouse body.This may be one of its dominant mechanism with delaying senility function.
Embodiment 48 pharmaceutical composition
Grinding and sieving, mixes and get final product.
Embodiment 49 pharmaceutical composition
Grinding and sieving, mixes and get final product.
Embodiment 50 pharmaceutical composition
Grinding and sieving, mixes and get final product.
Embodiment 51 pharmaceutical composition
Grinding and sieving, mixes and get final product.
Embodiment 52 pharmaceutical composition
Grinding and sieving, mixes and get final product.
Embodiment 53 pharmaceutical composition
Compound 3% shown in formula I
Dry starch 85%
Magnesium Stearate 12%
Grinding and sieving, mixes and get final product.
Embodiment 54 pharmaceutical composition
Compound 20% shown in formula I
Dry starch 65%
Magnesium Stearate 15%
Grinding and sieving, mixes and get final product.
Embodiment 55 pharmaceutical composition
Compound 17% shown in formula I
Dry starch 75%
Magnesium Stearate 8%
Grinding and sieving, mixes and get final product.
Embodiment 56 pharmaceutical composition
Compound 10% shown in formula I
Dry starch 85%
Magnesium Stearate 5%
Grinding and sieving, mixes and get final product.
Embodiment 57 pharmaceutical composition
Compound 15% shown in formula I
Dry starch 75%
Magnesium Stearate 10%
Grinding and sieving, mixes and get final product.
Embodiment 58 tablet
By compound shown in formula I, starch, Microcrystalline Cellulose, sodium starch glycolate and Magnesium Stearate, after mixing on tabletting machine compressing tablet, to obtain final product.
Embodiment 59 capsule
Compound 3% shown in formula I
Dry starch 85%
Magnesium Stearate 12%
By compound shown in formula I, dry starch and Magnesium Stearate, be packed into after mixing in hard gelatin capsule, obtain final product.
Embodiment 17 capsule
Compound 20% shown in formula I
Dry starch 65%
Magnesium Stearate 15%
By compound shown in formula I, dry starch and Magnesium Stearate, be packed into after mixing in hard gelatin capsule, obtain final product.
Embodiment 60 injection liquid
Compound shown in formula I is added ethanol stirring and dissolving, add sodium-chlor and appropriate water for injection, stir, add 0.1% pin activated carbon, absorption, filter decarburization, mend and inject water to specified amount, millipore filtration membrane filtration, embedding is propped up by 1mL/, 100 DEG C of moist heat sterilization 20min, qualified through lamp inspection, to obtain final product.
Embodiment 61 injection liquid
Compound shown in formula I is added ethanol stirring and dissolving, add sodium-chlor and appropriate water for injection, stir, add 0.1% pin activated carbon, absorption, filter decarburization, mend and inject water to specified amount, millipore filtration membrane filtration, embedding is propped up by 1mL/, 100 DEG C of moist heat sterilization 20min, qualified through lamp inspection, to obtain final product.
Embodiment 62 lyophilized injectable powder
Compound shown in formula I is added ethanol stirring and dissolving, adds sodium-chlor, N.F,USP MANNITOL and appropriate water for injection, stir, add 0.1% pin activated carbon, absorption, filters decarburization, mends and injects water to specified amount, millipore filtration membrane filtration, packing is propped up, lyophilize, encapsulation by 4mL/, through being up to the standards, to obtain final product.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (1)
1. a preparation method for formula I, is characterized in that, comprises following four steps:
The first step: get Breviscarpine raw material 28.0g, be placed in 1000mL round-bottomed flask, adds a certain amount of methyl alcohol 100g, DMF 400g and 0.56g calcium chloride, in 50 DEG C of heated and stirred reaction 2h;
Second step: reaction solution adopts vacuum concentration recycling design, when solution to be concentrated volume is reduced to about 1/3 of original volume, stop concentrated, concentrated solution leaves standstill and is cooled to room temperature, has a large amount of product crystallization;
3rd step: vacuum filtration, divide to take out for three times with 30mL cold methanol and wash, drain, products therefrom proceeds in 250mL ethyl acetate-light petrol, is heated to the 30min that boils as far as possible, sets to 0-5 DEG C of refrigerator and cooled but more than 4h;
4th step: take out crystallized product, vacuum filtration, takes out with 30mL cold ethyl acetate and wash three times, finally takes out with 15mL sherwood oil and washes once, drain as far as possible, put dry more than 48h in 60 DEG C of vacuum drying ovens, obtain the formula I of light yellow loose powders,
The volume ratio of described ethyl acetate and sherwood oil is 3:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210363570.8A CN102838645B (en) | 2012-09-26 | 2012-09-26 | Polyphenol hydroxy flavone compound with pharmaceutical function and preparation method thereof |
Applications Claiming Priority (1)
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