CN101019824B

CN101019824B - Preparation method of oral medicine composition containing salvianolic acid

Info

Publication number: CN101019824B
Application number: CN2007100909226A
Authority: CN
Inventors: 顾群; 李志刚; 渠守峰; 米长江; 栗艳彬; 林治荣; 鄯慧珍
Original assignee: BENCAO TIANYUAN PHARMACEUTICAL RESEARCH INST BEIJING
Current assignee: BENCAO TIANYUAN PHARMACEUTICAL RESEARCH INST BEIJING
Priority date: 2007-03-27
Filing date: 2007-03-27
Publication date: 2012-02-01
Anticipated expiration: 2027-03-27
Also published as: CN101019824A

Abstract

The present invention discloses one kind of oral medicine composition containing red sage salvianolic acid and its preparation process. The oral medicine composition has the composition determined through pharmacodynamic experiment, pharmacokinetic experiment and toxicologic experiment as red sage salvianolic acid 1-10 weight portions and breviscapine 1-20 weight portions, preferably red sage salvianolic acid 1-5 weight portions and breviscapine 1-5 weight portions. The present invention also discloses the detection and analysis method and the use of the medicine composition. Pharmacologic experiment shows its excellent pharmacologic effect.

Description

A kind of method for preparing that contains the combination of oral medication of danshen root salvianolic acid A

Technical field

The present invention relates to technical field of traditional Chinese medicine pharmacy, be specifically related to a kind of combination of oral medication that contains danshen root salvianolic acid A and preparation method thereof.

Background technology

Salvianolic acid A [salvianolic acid A] is a strongest active composition in the Radix Salviae Miltiorrhizae, has great pharmacological effects (Du Guanhua [preclinical medicine and clinical, 2000; 20 (5): 10～14], Hu Yiyang [herbal pharmacology journal, 1997,18 (5): 478-480]); Danshen root salvianolic acid A content in Radix Salviae Miltiorrhizae is very low, has only about 5/10000ths, even through a series of technology it is extracted; Can only obtain the salvianolic acid A of trace level; Therefore Chinese medicine carries out compatibility with Radix Salviae Miltiorrhizae and other medical material, its essence is the compatibility with the more weak composition (such as TANSHINONES, salvianolic acid B etc.) of other relativity of Radix Salviae Miltiorrhizae, though this compatibility has played certain therapeutical effect clinically; But the effective ingredient with the maximum drug effect of performance in the Radix Salviae Miltiorrhizae is not applied in the treatment of diseases, has caused the huge waste of resource.Therefore the salvianolic acid A of level is applied to the hope that treatment of diseases is a lot of medical scientific research persons in batches, the compatibility of the salvianolic acid A of level and pharmaceutically active ingredient in other in batches particularly, one of difficult problem that especially need be to be solved.

Breviscapine is the flavonoids effective constituent that from Compositae Herba Erigerontis aceris plant Herba Erigerontis (Erigeron brieviscapus (Vant) Hand Mazz.), extracts; Have expansion of cerebral vascular, arteria coronaria blood vessel and peripheral vascular effect; Blood flow is increased; Improve local blood supply, make Peripheral resistance descend microcirculation improvement etc.; The preparation that with the breviscapine is raw material clinically includes Herba Erigerontis sheet etc., and these preparations have certain therapeutical effect in clinical practice to thrombosis property cardiovascular and cerebrovascular disease, but thrombosis dissolves when loosing the recovery organization OBF; Generation reperfusion injury that will be in various degree can cause even more serious consequence, and the breviscapine oral administration preparation can't solve the produced simultaneously reperfusion injury of thrombolytic preferably; Thereby its curative effect is not very desirable, need be in clinical the compatibility several drugs, to reduce the damage of perfusion generation again behind the thrombolytic; These simple clinically compatibilities though can reach certain therapeutical effect, can produce a large amount of untoward reaction; Bring huge medication difficulty to the doctor, bring bigger misery to the patient, therefore; Should carry out a series of experiment, the combination of pharmaceutically active ingredient or component effective site in the excavation to the compatibility of middle pharmaceutically active ingredient or component effective site; To reduce untoward reaction, increase curative effect, can further promote the process of the modernization of Chinese medicine.

The research of Chinese medicine compatibility rests on the basis of medical material compatibility more, though can make an explanation with theory of Chinese medical science, there is dispute in its science always; Therefore middle pharmaceutically active ingredient or component effective site are carried out compatibility, by numerous researcher approvals, this compatibility and proportion relation are its core and soul place; Be the characteristic and the advantage of Chinese medicine; It neither simple Chinese medicine medical material use, quantitative addition, the counteracting of toxic reaction that neither machinery, but through a series of research (aspects such as medicine efficacy screening, system's pharmacodynamics evaluation and pharmacokinetics); On the basis of dialectical method; With legally constituted authority side, cube send medicine, the organic teacher who combines in order.

Consult document and patent; The report that danshen root salvianolic acid A and breviscapine compatibility are not arranged; But the compatibility that Radix Salviae Miltiorrhizae total phenolic acids and breviscapine are arranged, though this drug regimen has certain curative effect because prior art exist certain defective: obtaining salvia miltiorrhiza tanshinoate is to be main salvianolic acid class material with the salvianolic acid B of Radix Salviae Miltiorrhizae; And can't obtain the danshen root salvianolic acid A of level in batches; Therefore, can the danshen root salvianolic acid A and the breviscapine of level make up in batches, and needing the scientific research personnel to pass through a large amount of scientific experiments could confirm.

Summary of the invention

For these reasons; We are about to Radix Salviae Miltiorrhizae total phenolic acids and have obtained the danshen root salvianolic acid A of level, the mode of administered through oral administration more in batches through certain chemical reaction through red rooted salvia is carried out a series of Study on Transformation; Confirmed that with experiments such as medicine efficacy screening, system's drug effect and safety evaluatio, pharmacokineticss a certain amount of danshen root salvianolic acid A and a certain amount of breviscapine can make up; This drug regimen can also well solve the reperfusion injury that produces behind the thrombolytic except having complementary effect, reduced the untoward reaction of clinical drug combination; Having reached 1+1 promptly increases curative effect greater than 2, reduces the effect of untoward reaction.

The application realizes through following proposal.

The combination of oral medication that contains danshen root salvianolic acid A, wherein: danshen root salvianolic acid A 1-10 weight portion, breviscapine 1-20 weight portion;

Contain the combination of oral medication of danshen root salvianolic acid A, wherein be preferably: danshen root salvianolic acid A 1-5 weight portion, breviscapine 1-5 weight portion;

Wherein breviscapine content in lamp-dish flower acetic more than or equal to 50% and less than 100%;

Wherein the content of danshen root salvianolic acid A is more than or equal to 50% and less than 100%;

The drug oral preparation of compositions becomes tablet, capsule, granule, soft capsule, pellet, drop pill, the oral liquid of UD;

Wherein UD is 20mg-4000mg; Wherein preferred unit dosage is 50-2000mg.UD is lower than 20mg does not have effect in clinical use, UD can produce certain toxicity greater than 4000mg in clinical use.

The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents the application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.

One. process recipes

Danshen root salvianolic acid A extracts purification:

Radix Salviae Miltiorrhizae water or alcoholic solution extract and obtain aqueous extract or alcohol extract; Alcohol extract concentrates ethanol to most; Transfer pH value to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value, heated 1-6 hour to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure; Solution filters, and filtrating is separated through nonpolar or low pole macroporous resin column chromatography, and macroporous resin column is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8; Elder generation's water, 10-30% Diluted Alcohol eluting are removed impurity, the ethanol elution of reuse 30-70% concentration; Collect eluent; Concentrate, drying obtains danshen root salvianolic acid A;

Or:

Radix Salviae Miltiorrhizae water or alcoholic solution extract and obtain aqueous extract or alcohol extract; Alcohol extract concentrates ethanol to most; Transfer pH value to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value, heated 1-6 hour to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure; Solution filters; Filtrating is separated through nonpolar or low pole macroporous resin column chromatography, and macroporous resin column is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8, first water, 10-30% Diluted Alcohol eluting; Remove impurity; The ethanol elution of reuse 30-70% concentration is collected eluent, reclaims ethanol to most; Concentrated solution separates with sephadex lh-20 or polyamide column chromatography, and first water, 20-50% alcoholic solution eluting discard eluent, and reuse 50-95% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A;

Or:

Radix Salviae Miltiorrhizae water or alcoholic solution extract and obtain aqueous extract or alcohol extract; Alcohol extract concentrates ethanol to most; Transfer pH value to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value, heated 1-6 hour to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure; Solution filters; Filtrating is separated through nonpolar or low pole macroporous resin column chromatography, and macroporous resin column is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8, first water, 10-30% Diluted Alcohol eluting; Remove impurity; The ethanol elution of reuse 30-70% concentration is collected eluent, reclaims ethanol to most; Concentrated solution separates with sephadex lh-20 or polyamide column chromatography, and first water, 20-50% alcoholic solution eluting discard eluent, and reuse 50-95% alcoholic solution eluting reclaims ethanol to most; Concentrated solution transfers pH value to 2-5, and through organic solvent extraction, organic solvent is selected from a kind of in ethyl acetate, propyl acetate, butyl acetate, n-butyl alcohol, the isopropyl alcohol, separates the organic solvent phase, must contain drug solns, concentrate, drying or lyophilization, danshen root salvianolic acid A;

Breviscapine is a prior art for preparing, wherein breviscapine content in lamp-dish flower acetic more than or equal to 50% and less than 100% (commercially available or extract purification obtain) according to literature method;

Formulation preparation:

Preparation tablets:

Get danshen root salvianolic acid A and breviscapine, be prepared into tablet according to the conventional requirement of tablet pharmaceutics;

The capsule preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into capsule according to the conventional requirement of the pharmaceutics of capsule;

The granule preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into granule according to the conventional requirement of the pharmaceutics of granule;

The soft capsule preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into soft capsule according to the conventional requirement of the pharmaceutics of soft capsule;

The pellet preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into pellet according to the conventional requirement of the pharmaceutics of pellet;

The drop pill preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into drop pill according to the conventional requirement of the pharmaceutics of drop pill;

The oral liquid preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into oral liquid according to the conventional requirement of the pharmaceutics of oral liquid;

Two. the check and analysis method

1. detection and analytic method for red sage root dan phenolic acid A:

Chromatographic column: C ₁₈Reversed phase chromatographic column, NUCLEODUR, 250*4.6mm, ODS;

Chromatographic condition and system suitability experiment: with the octadecylsilane chemically bonded silica is filler; Flow velocity 1.0ml/min; 35 ℃ of column temperatures; Detect wavelength 286nm; With acetonitrile-0.2% aqueous acetic acid is mobile phase, carries out gradient elution by following condition of gradient elution, moves 90 minutes;

In the time of 0-15 minute, the ratio of acetonitrile by 10% rise to 20%, 0.2% aqueous acetic acid ratio reduce to 80% by 90%; In the time of 15-55 minute, the ratio of acetonitrile by 20% rise to 30%, 0.2% aqueous acetic acid ratio reduce to 70% by 80%; In the time of 55-65 minute, the ratio of acetonitrile by 30% rise to 50%, 0.2% aqueous acetic acid ratio reduce to 50% by 70%; In the time of 65-72 minute, the ratio of acetonitrile by 50% rise to 80%, 0.2% aqueous acetic acid ratio reduce to 20% by 50%; In the time of 72-77 minute, the ratio 20% of ratio 80%, 0.2% aqueous acetic acid of acetonitrile; In the time of 77-80 minute, the ratio of acetonitrile by 80% reduce to 10%, 0.2% aqueous acetic acid ratio rise to 90% by 20%; In the time of 80-90 minute, keep acetonitrile-0.2% aqueous acetic acid to carry out eluting with 10: 90 ratio;

The preparation of reference substance solution: precision takes by weighing the salvianolic acid A reference substance in volumetric flask, adds dissolve with methanol and shakes up, and be diluted to scale;

The preparation of sample solution: precision takes by weighing the salvianolic acid A sample, adds dissolve with methanol and shakes up, and be diluted to scale; Or precision measures or takes by weighing preparation, adds dissolve with methanol and shakes up, and be diluted to scale;

Algoscopy: accurate respectively reference substance solution and the sample solution drawn, inject chromatograph of liquid, the record chromatogram adopts external standard method with calculated by peak area, promptly gets;

2. breviscapine check and analysis:

Chromatographic condition and system suitability experiment: with the octadecylsilane chemically bonded silica is filler; Flow velocity 1.0ml/min; 30 ℃ of column temperatures; Detect wavelength 335nm; Number of theoretical plate is not less than 2000 by lamp-dish flower acetic; With volume ratio methanol: glacial acetic acid: water is that 32: 2: 66 solution is mobile phase;

The preparation of reference substance solution: precision takes by weighing the lamp-dish flower acetic reference substance, adds dissolve with methanol and shakes up, and be diluted to scale;

The preparation of need testing solution: precision takes by weighing breviscapine, adds dissolve with methanol and shakes up, and be diluted to scale; Or precision takes by weighing or measures preparation, adds dissolve with methanol and shakes up, and be diluted to scale;

Experimental result is seen table 1, table 2:

The check and analysis of table 1 crude drug

Group	Content
		Danshen root salvianolic acid A %	≥50，＜100
Breviscapine (lamp-dish flower acetic meter) %	≥50，＜100

The check and analysis of table 2 preparation

Group	Salvianolic acid A content	Breviscapine content (in lamp-dish flower acetic)
			The application's tablet (salvianolic acid A, breviscapine are raw material) mg/ sheet the application capsule (salvianolic acid A, breviscapine are raw material) mg/ grain the application granule (salvianolic acid A, breviscapine are raw material) mg/ bag the application soft capsule (salvianolic acid A, breviscapine are raw material) mg/ grain the application pellet (salvianolic acid A, breviscapine are raw material) mg/10 ball the application drop pill agent (salvianolic acid A, breviscapine are raw material) mg/10 ball	40-400 40-400 40-400 40-400 40-400 40-400	40-800 40-800 40-800 40-800 40-800 40-800
The application's oral solutions (salvianolic acid A, breviscapine are raw material) mg/ bottle	40-400	40-800

Experiment conclusion: show that through above-mentioned experiment the application's drug regimen has practical significance.

Three. the pharmacodynamics confirmatory experiment

1. to the protective effect of anesthetized rat myocardial ischemia reperfusion injury

Experimental program:

Scheme 1: danshen root salvianolic acid A 1 weight portion, breviscapine 1 weight portion;

Scheme 2: danshen root salvianolic acid A 1 weight portion, breviscapine 20 weight portions;

Scheme 3: danshen root salvianolic acid A 1 weight portion, breviscapine 5 weight portions;

Scheme 4: danshen root salvianolic acid A 5 weight portions, breviscapine 1 weight portion;

Scheme 5: danshen root salvianolic acid A 4 weight portions, breviscapine 3 weight portions;

Scheme 6: danshen root salvianolic acid A 7 weight portions, breviscapine 19 weight portions;

Scheme 7: danshen root salvianolic acid A 10 weight portions, breviscapine 20 weight portions;

Scheme 8: danshen root salvianolic acid A 5 weight portions;

Scheme 9: breviscapine 5 weight portions;

Experimental technique:

Get the healthy SD rat, body weight 240-260g, random packet: blank group, experimental program group.Place equivalent environment to raise 2 days free diet in advance.After raising end in advance, experimentize, animal is weighed, and 20% urethane is pressed the 0.6ml/100g lumbar injection; After treating that anesthesia is satisfied, lie on the back and be fixed on the Mus plate, tracheal intubation connects respirator; By 10～12ml tidal volume, 70 times/minute frequency is exhaled, and continuous positive pressure breathing is inhaled: exhale than being 1: 1.According to respiratory frequency and degree of depth adjustment respiration parameter.Connect electrocardiograph subsequently, survey normal ECG.Cut off front field of operation hair, iodine disinfection is cut off skin, subcutaneous tissue, front muscle and fascia 3～4cm, and it is long to separate Intercostal muscle 3cm with 1 8# vascular forceps along the 3rd intercostal passivity; Open thoracic cavity and pericardium, recording ecg struts 3,4 ribs; Refer to hold thoracic cavity, rat right side with left hand four, the assistant upwards pushes away thymus with the ophthalmology tweezer, between left auricle and pulmonary conus, finds ligation sign blood vessel great cardiac vein; The 2mm place is with noinvasive roundlet pin band 6-0 silk thread threading below left auricle, and depth of needle is 1～1.5mm, wide 2～3mm; Recording ecg behind the threading, gastric infusion (dosage is 24mg/kg), the blank group gives the normal saline of respective amount; Recording ecg behind the administration 10min, and with one the band groove little plastics pipe pad at the ligation position, the ligation above that of two rear line heads.At once recording ecg after the ligation is cyanosis or the II S-T section back of a bow that leads with left chamber antetheca and upwards raises greater than 0.1mv and be that ligation successfully indicates (it is superseded that the S-T section does not have the changer) more than the lasting 0.5h.10min recording ecg is once more cut off ligature behind the ligation 30min after the ligation, realizes perfusion again, and record pours into electrocardiogram at once again, removes in the thoracic cavity layer-by-layer suture thoracic wall behind the hematocele, removes respirator, animal recovery autonomous respiration, and incision of trachea does not process.Irritate again at once, 10min, 20min, 40min, 1h, 2h, 3h recording ecg respectively.Irritated 3 hours, through abdominal aortic blood, 4000rpm is centrifugal, and 10min gets serum again, and dissection is cored dirty, and the residual blood of ice normal saline flush away cuts off atrium and right ventricle, puts into refrigerator and cooled immediately and freezes.With heart after refrigerator and cooled is frozen 10min, from the apex of the heart entad the parallel coronary sulcus direction in the end 5 of equal thickness are cut in left chamber, put into the 1%TTC dye liquor, 37 ℃ of dyeing 10min, the necrotic area is not a kermesinus, the necrotic area is canescence.Digital camera is taken pictures.Weighed respectively in necrotic area and non-necrotic area, calculate the percentage ratio that the necrotic area accounts for left ventricular mass, i.e. infarction size.

The detection index is respectively, myocardial infarct size.Experimental result sees table 3 for details:

The influence

of myocardial ischemia myocardial infarct size (%) due to table 3 pair ligation/logical again rat ramus descendens anterior arteriae coronariae sinistrae

Group	Myocardial infarct size (%)
		Matched group diltiazem hydrochloride injection scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8	26.02±11.92 8.47±2.59 ^ 11.23±1.17 ^# 13.62±1.42 ^# 11.68±1.27 ^# 11.27±1.57 ^# 11.54±1.10 ^# 13.31±1.35 ^# 13.62±1.34 ^# 16.15±4.03 ^*
Scheme 9	23.77±4.85

Annotate: compare with matched group ^*P＜0.01, ^*P＜0.05; Compare #P＜0.05 with scheme 9.

2. rat lipid-lowering test

Experimental program:

Scheme 1: danshen root salvianolic acid A 10 weight portions, breviscapine 1 weight portion;

Scheme 2: danshen root salvianolic acid A 10 weight portions, breviscapine 4 weight portions;

Scheme 3: danshen root salvianolic acid A 10 weight portions, breviscapine 18 weight portions;

Scheme 4: danshen root salvianolic acid A 1 weight portion, breviscapine 3 weight portions;

Scheme 5: danshen root salvianolic acid A 3 weight portions, breviscapine 5 weight portions;

Scheme 6: danshen root salvianolic acid A 7 weight portions, breviscapine 3 weight portions;

Scheme 7: danshen root salvianolic acid A 5 weight portions, breviscapine 5 weight portions;

Scheme 8: danshen root salvianolic acid A 5 weight portions;

Scheme 9: breviscapine 5 weight portions;

Experimental technique: male SD rat, adaptability were fed after the week, were divided into normal control group, model group, experimental program group at random by body weight.Except that the distilled water of normal control group every mornings 8 point-9 a filling stomach 1.0ml/100g; All the other groups are irritated stomach 1.0ml/100g lipomul; Prescription is 30% Adeps Sus domestica, 10% cholesterol, 2% cholate, 0.2% propylthiouracil, 20% Tween 80,20% propylene glycol, 10% sucrose; With distilled water melt into 100ml, continuous 10 days.12 hours socket of the eye venous plexuses of fasting are got blood, detect serum TC, TG, HDL-C and LDL-C content, and divide into groups again by the serum TC level.After adjustment is divided into groups; Group rat every morning gives lipomul by above-mentioned modeling test method; Emulsion formulations becomes: 20% Adeps Sus domestica, 5% cholesterol, 2% cholate, 0.2% propylthiouracil, 10% tween, 10% propylene glycol, 5% sucrose are with distilled water melt into 100ml; Afternoon 1, group dosage gastric infusion was pressed in point-2, and in 4 weeks of successive administration, dosage is 24mg/kg.After 4 weeks of administration finish, fasting 12 hours, femoral artery is got blood, and a part of blood is used for hemorheology and detects with 1% anticoagulant heparin (about 4ml).Another part blood separation serum is used for the detection of biochemical indicator.Duration of test is claimed body weight weekly one time, by body weight adjustment dosage, and measures food ration.4 weeks of administration are detected serum TC, TG, LDL-C and HDL-C level respectively, and experimental result is seen table 4

Table 4 administration 4 all red lamps are to the influence

of rat fat

Group	TC (mmol/L)	TG (mmol/L)	HDL-C (mmol/L)	LDL-C (mmol/L)
					Normal group model group scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8	1.89±0.27 ^ 16.26±5.30 10.25±2.05 ^# 10.31±2.28 ^# 9.77±2.21 ^# 10.41±2.22 ^ 10.55±2.29 ^ 10.28±2.61 ^ 10.24±2.74 ^ 13.98±3.80 ^*	1.04±0.17 ^** 0.55±0.17 0.78±0.23 ^* 0.77±0.24 ^* 0.82±0.21 ^** 0.75±0.19 ^* 0.79±0.18 ^* 0.72±0.18 ^* 0.75±0.18 ^* 0.68±0.18	0.79±0.11 ^** 1.97±0.24 1.29±0.11 ^* 1.30±0.10 ^* 1.20±0.09 ^* 1.33±0.12 ^* 1.35±0.10 ^* 1.38±0.09 ^* 1.37±0.13 ^* 1.57±0.21	0.61±1.17 ^ 6.21±1.60 4.23±0.70 ^# 4.27±0.72 ^# 3.92±0.78 ^# 4.30±0.80 ^# 4.28±0.84 ^# 4.24±0.89 ^# 4.17±1.03 ^# 5.47±1.69 ^*
Scheme 9	13.94±4.04 ^*	0.67±0.20	1.52±0.20	5.98±1.71 ^*

Annotate: compare with matched group ^*P＜0.01, ^*P＜0.05; Compare #P＜0.05 with scheme 8,9.

3. cerebrovascular disease pharmacological evaluation

Experimental program:

Scheme 1: danshen root salvianolic acid A 1 weight portion, breviscapine 3 weight portions;

Scheme 2: danshen root salvianolic acid A 1 weight portion, breviscapine 5 weight portions;

Scheme 3: danshen root salvianolic acid A 8 weight portions, breviscapine 6 weight portions;

Scheme 4: danshen root salvianolic acid A 2 weight portions, breviscapine 9 weight portions;

Scheme 5: danshen root salvianolic acid A 4 weight portions, breviscapine 8 weight portions;

Scheme 6: danshen root salvianolic acid A 5 weight portions, breviscapine 20 weight portions;

Scheme 7: danshen root salvianolic acid A 9 weight portions, breviscapine 19 weight portions;

Scheme 8: danshen root salvianolic acid A 5 weight portions;

Scheme 9: breviscapine 5 weight portions;

Experimental technique: adopt middle cerebral artery occlusion (MACO) to induce the method for focal cerebral ischemia in rats model, inquire into the preventive and therapeutic effect of different schemes to local cerebral ischemia due to the intraluminal middle cerebral artery occlusion in rats obturation.Use normal saline as negative control group, the positive contrast of nimodipine, gastric infusion, dosage is 24mg/kg, and scheme group dosage is 24mg/kg, contains aquametry as index with brain, and experimental result is seen table 5:

Table 5 pair MACO rat cerebral tissue water content

Group	Left hemisphere brain water content %	Right hemisphere brain water content %
			Matched group nimodipine prescription case 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8	81.17±0.89 77.34±0.44 77.92±0.52 77.97±0.47 77.80±0.51 77.95±0.50 77.91±0.48 77.97±0.53 77.99±0.44 77.91±0.47	83.95±0.92 77.52±0.48 ^ 79.17±0.47 ^# 79.88±0.51 ^# 79.77±0.59 ^# 80.28±0.54 ^# 80.57±0.61 ^# 80.26±0.42 ^# 80.74±0.50 ^# 81.34±0.54 ^*
Scheme 9	77.97±0.46	81.75±0.59 ^*

Annotate: compare with matched group ^*P＜0.01, ^*P＜0.05; Compare #P＜0.05 for 18 groups with scheme 8, scheme.

4. anti-ageing year dementia pharmacological evaluation

Experimental program:

Scheme 1: danshen root salvianolic acid A 5 weight portions, breviscapine 3.5 weight portions;

Scheme 2: danshen root salvianolic acid A 9 weight portions, breviscapine 5.5 weight portions;

Scheme 4: danshen root salvianolic acid A 6 weight portions, breviscapine 18 weight portions;

Scheme 5: danshen root salvianolic acid A 6 weight portions, breviscapine 17 weight portions;

Scheme 6: danshen root salvianolic acid A 3 weight portions, breviscapine 14 weight portions;

Scheme 7: danshen root salvianolic acid A 5 weight portions, breviscapine 1 weight portion;

Scheme 8: danshen root salvianolic acid A 5 weight portions;

Scheme 9: breviscapine 5 weight portions;

Experimental technique: the normal rat of the diving tower method of learning from else's experience and the screening of eight arm maze methods adopts intracerebroventricular injection β-AP _25-35, process Alzheimer (AD) rat model, use space learning and memory ability that diving tower method and eight arms electricity maze method is judged administration front and back rat; Adopt chemical colorimetry to measure Acetylcholinesterase (AchE) activity in the cerebral tissue, use normal saline as negative control group, scheme group gastric infusion amount is 24mg/kg.

Experimental result: compare with the negative control group rat, rat model eight arms electricity labyrinth errors number obviously increases (P＜0.05), and diving tower learning and memory errors number obviously increases (P＜0.01).Continuous quiet notes administration is after 21 days before and after the rat modeling, the above-mentioned behavioristics of scheme 1-7 group rat index be improved significantly (P＜0.01), active reduce (P＜0.01) of cerebral tissue AchE.The result shows that the application's preparation is to β-AP _25-35Rat space learning and dysmnesia have significant prevention and therapeutical effect due to the intracerebroventricular injection, and the AchE activity is dependency in this effect and the reduction cerebral tissue.Other group effect is not too obvious, does not have significant difference with model group.

5. anti-organ fibrosis experiment

Scheme 1: danshen root salvianolic acid A 1 weight portion, breviscapine 4 weight portions;

Scheme 2: danshen root salvianolic acid A 5.5 weight portions, breviscapine 15.5 weight portions;

Scheme 3: danshen root salvianolic acid A 3 weight portions, breviscapine 6 weight portions;

Scheme 4: danshen root salvianolic acid A 9 weight portions, breviscapine 13 weight portions;

Scheme 5: danshen root salvianolic acid A 2 weight portions, breviscapine 7 weight portions;

Scheme 6: danshen root salvianolic acid A 4 weight portions, breviscapine 14 weight portions;

Scheme 7: danshen root salvianolic acid A 3 weight portions, breviscapine 2 weight portions;

Scheme 8: danshen root salvianolic acid A 5 weight portions;

Scheme 9: breviscapine 5 weight portions;

Experimental technique: cause the rat liver fibrosis model with 40% carbon tetrachloride complex factors.Give simultaneously and the treatment of scheme group, scheme group gastric infusion, dosage is 24mg/kg, uses normal saline as negative control group.Detect liver function, III procollagen type (pcIII), hyaluronic acid (HA) during off-test, separate hepatic tissue and detect liver hydroxyproline content and the variation of electron microscopic observation hepatic tissue pathology.

Experimental result: scheme 1-7 group can obviously be improved the liver function of hepatic fibrosis rats, reduces serum pCIII, HA content and the hepatic tissue hydroxyproline is obviously descended; Obviously alleviate the deposition of fat-storing cell hypertrophy and collagen.Other group effect is not too obvious, does not have significant difference with negative control group.

6. microcirculation pharmacological evaluation

Experimental program:

Scheme 1: danshen root salvianolic acid A 5 weight portions, breviscapine 1 weight portion;

Scheme 2: danshen root salvianolic acid A 1 weight portion, breviscapine 6.5 weight portions;

Scheme 3: danshen root salvianolic acid A 2 weight portions, breviscapine 5 weight portions;

Scheme 4: danshen root salvianolic acid A 1 weight portion, breviscapine 5 weight portions;

Scheme 5: danshen root salvianolic acid A 9 weight portions, breviscapine 4 weight portions;

Scheme 6: danshen root salvianolic acid A 1 weight portion, breviscapine 7 weight portions;

Scheme 7: danshen root salvianolic acid A 5 weight portions, breviscapine 15 weight portions;

Scheme 8: danshen root salvianolic acid A 5 weight portions;

Scheme 9: breviscapine 5 weight portions;

Experimental technique: micro tv amplification system quantitative observation scheme group is to auricular microcirculation obstacle microcirculation of mouse auricle influence due to normal and the norepinephrine (NA); The multiple CAL of blood plasma is measured anticoagulation, scheme group gastric infusion, and dosage is 36mg/kg.

Experimental result: scheme 1-7 group can significantly promote or improve the microcirculation that normally reaches auricular microcirculation obstacle Mice Auricle due to the NA; Also can prolong the blood plasma recalcification time.

7. anti-tumor experiment

Experimental program:

Scheme 1: danshen root salvianolic acid A 5 weight portions, breviscapine 3 weight portions;

Scheme 2: danshen root salvianolic acid A 2 weight portions, breviscapine 5 weight portions;

Scheme 3: danshen root salvianolic acid A 8 weight portions, breviscapine 16 weight portions;

Scheme 4: danshen root salvianolic acid A 5 weight portions, breviscapine 9 weight portions;

Scheme 5: danshen root salvianolic acid A 6 weight portions, breviscapine 3 weight portions;

Scheme 6: danshen root salvianolic acid A 2 weight portions, breviscapine 17 weight portions;

Scheme 7: danshen root salvianolic acid A 3 weight portions, breviscapine 20 weight portions;

Scheme 8: danshen root salvianolic acid A 5 weight portions;

Scheme 9: breviscapine 5 weight portions;

Experimental technique: with the antimutagenic effect of mouse Bone marrow cells micronucleus experiment and testicular chromosome distortion laboratory observation scheme group, with the antitumous effect of S-180 and H-22 transplanted tumor observation plan group, scheme group gastric infusion, dosage is 36mg/kg.

Experimental result: scheme 1-7 group all has the obvious suppression effect to the mouse testis cell chromosome that mouse Bone marrow cells micronucleus takes place and mitomycin the brings out distortion that cyclophosphamide brings out; S-180 and the growth of H-22 mice transplanted tumor also there is the obvious suppression effect.Show that scheme 1-7 group all has protective effect to the DNA damage of somatic cell and sexual cell, also has certain tumor-inhibiting action to mice transplanted tumor.

Experiment 8

The pharmacological evaluation of treatment gout

Experimental program:

Scheme 1: danshen root salvianolic acid A 1 weight portion, breviscapine 5 weight portions;

Scheme 2: danshen root salvianolic acid A 5 weight portions, breviscapine 1 weight portion;

Scheme 3: danshen root salvianolic acid A 2 weight portions, breviscapine 4 weight portions;

Experimental technique: uric acid is dissolved in the NaOH solution of 1mol/L,,, is prepared into aseptic urate crystal through stirring, filtration, cold drying, screening, sterilization with HCl adjustment PH.With aseptic manipulation, with No. 4 injection needles,, inject the articulatio tibiotarsalis intracavity of gout model group and scheme group rat right hind leg internal malleolus with 5% urate solution, 50 μ l, each animal is only injected once.Scheme group rat in gout model preparation, gastric infusion, dosage 24mg/kg, for three days on end, normal group and gout model group are given the equal-volume normal saline.Detect ankle joint and surrounding tissue K thereof ⁺Concentration, experimental result is seen table 6:

The K of table 6 ankle joint and surrounding tissue thereof ⁺Concentration

Group	Number of animals only	K ⁺ C _n/mmol*L ^-1
			2 groups of normal group gout model group scheme 1 prescription cases	10 10 10 10	0.89±0.07 1.28±0.11 1.10±0.09 ^ 1.07±0.08 ^
3 groups of schemes	10	1.08±0.08 ^**

Annotate: compare with the gout model group ^*P＜0.01

Pharmacological evaluation brief summary: show that through above-mentioned pharmacological evaluation drug regimen and matched group have great pharmacological effects (P＜0.01) in the application's scope; Relatively have significant difference (P＜0.05) with danshen root salvianolic acid A, breviscapine, prove absolutely the combination of danshen root salvianolic acid A and breviscapine except that having good complementary action, also having interacts improves the effect of pharmacologically active.

Four. pharmacokinetic

Experimental program:

Scheme 1: danshen root salvianolic acid A 1 weight portion, breviscapine 21 weight portions;

Scheme 2: danshen root salvianolic acid A 1 weight portion, breviscapine 3 weight portions;

Scheme 3: danshen root salvianolic acid A 1 weight portion, breviscapine 4 weight portions;

Scheme 5: danshen root salvianolic acid A 2 weight portions, breviscapine 4 weight portions;

Scheme 6: danshen root salvianolic acid A 1 weight portion, breviscapine 22 weight portions;

Scheme 8: danshen root salvianolic acid A 4 weight portions, breviscapine 1 weight portion;

Scheme 9: danshen root salvianolic acid A 3 weight portions, breviscapine 2 weight portions;

Scheme 10: danshen root salvianolic acid A 11 weight portions, breviscapine 1 weight portion;

Scheme 11: danshen root salvianolic acid A 6 weight portions, breviscapine 19 weight portions;

Scheme 12: danshen root salvianolic acid A 5 weight portions, breviscapine 20 weight portions;

Scheme 13: danshen root salvianolic acid A 9 weight portions, breviscapine 1 weight portion;

Scheme 14: danshen root salvianolic acid A 2 weight portions, breviscapine 19 weight portions;

Scheme 15: danshen root salvianolic acid A 8 weight portions, breviscapine 1 weight portion;

Scheme 16: danshen root salvianolic acid A 12 weight portions, breviscapine 2 weight portions;

Scheme 17: danshen root salvianolic acid A 2 weight portions, breviscapine 23 weight portions;

Scheme 18: danshen root salvianolic acid A 5 weight portions;

Scheme 19: breviscapine 5 weight portions;

Laboratory animal: the SD rat, male, 8 ages in week, SPF level; Assay method: select for use the LC-MS/MS method to detect; The plasma treatment method: precision is measured plasma specimen 100 μ l, adds 100 μ l, 0.1% phosphoric acid, 400 μ l methanol, behind the vortex mixing, and the centrifugal 5min of 10000r/min.After 0.2 μ m filter filtered, supernatant was through the automatic sampler sample introduction; Male SD rat; Through gastric infusion, get blank blood before the medicine, in taking medicine back 15min, 30min, 45min, 1h, 1.5h, 2h, 3h, 4h, 6h, 8h and 12h extracting vein blood 0.3ml in anticoagulant tube; Separated plasma; With danshen root salvianolic acid A and breviscapine is that the ejection preparation that feedstock production becomes is a benchmark, calculates the different schemes absolute bioavailability, and experimental result is seen table 7:

The investigation of table 7 different schemes bioavailability

Group	F salvianolic acid A %	F breviscapine %
			Scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8 schemes 9 schemes 10 schemes 11 schemes 12 schemes 13 schemes 14 schemes 15 schemes 16 schemes 17 schemes 18	12.9 14.9 16.7 17.2 18.4 12.3 18.1 17.3 17.8 12.2 16.5 17.8 17.4 18.1 17.3 12.3 12.0 11.8	0.44 0.60 0.67 0.66 0.72 0.49 0.85 0.79 0.72 0.46 0.82 0.73 0.78 0.79 0.69 0.47 0.45 -
Scheme 19	-	0.41

Five. toxicologic study

Experimental program:

Scheme 2: danshen root salvianolic acid A 5 weight portions, breviscapine 3 weight portions;

Scheme 3: danshen root salvianolic acid A 5 weight portions, breviscapine 1 weight portion;

Scheme 4: danshen root salvianolic acid A 4 weight portions, breviscapine 2 weight portions;

Scheme 5: danshen root salvianolic acid A 7 weight portions, breviscapine 3 weight portions;

Scheme 6: danshen root salvianolic acid A 8 weight portions, breviscapine 1 weight portion;

Scheme 7: danshen root salvianolic acid A 10 weight portions, breviscapine 17 weight portions;

Scheme 8: danshen root salvianolic acid A 5 weight portions, breviscapine 20 weight portions;

Scheme 9: danshen root salvianolic acid A 11 weight portions, breviscapine 1 weight portion;

Scheme 10: danshen root salvianolic acid A 5 weight portions;

Scheme 11: breviscapine 5 weight portions;

Experimental technique: the pharmaceutical composition of above-mentioned different schemes, carry out toxicological experiment, measure the LD of mice oral administration acute toxicity ₅₀, experimental result is seen table 8:

The LD of table 8 different schemes ₅₀Value

Group	LD ₅₀Value g/kg
		Scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8 schemes 9 schemes 10	17.1 20.3 20.7 19.8 19.7 18.4 19.2 20.1 15.8 15.4
Scheme 11	12.91

Experiment conclusion: through pharmacodynamics experiments, pharmacokinetics experiment, toxicological experiment, we confirm danshen root salvianolic acid A 1-10 weight portion, breviscapine 1-20 weight portion; Preferred danshen root salvianolic acid A 1-5 weight portion, breviscapine 1-5 weight portion.

Six. preparation embodiment

Embodiment 1

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae extracts with 85% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 9.0,80 ℃ of temperature, heating 6 hours, and solution filters; Filtrating is separated through the HPD-400 macroporous resin column chromatography, and first water, 30% Diluted Alcohol eluting are removed impurity; The ethanol elution of reuse 70% concentration is collected eluent, concentrates; Drying obtains danshen root salvianolic acid A, danshen root salvianolic acid A content 59.3%.

Or

Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transfers pH value to 3.5,110 ℃ of temperature, gauge pressure 0.05MPa pressure, heats 6 hours; Solution filters, and filtrating is separated through the HPD-400A macroporous resin column chromatography, and first water, 30% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 70% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with polyamide chromatography post, and first water, 50% alcoholic solution eluting discard eluent, and reuse 95% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, danshen root salvianolic acid A content 89.7%

Or

Radix Salviae Miltiorrhizae extracts with 80% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 6.0,130 ℃ of temperature, gauge pressure 0.17MPa pressure, heats 6 hours; Solution filters, and filtrating is separated through the HPD-450 macroporous resin column chromatography, and first water, 30% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 70% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 50% alcoholic solution eluting discard eluent, and reuse 95% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 5, through the extraction of organic solvent propyl acetate, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, danshen root salvianolic acid A content 99.4%.

Breviscapine is a prior art for preparing, and its content counts 99.1% with lamp-dish flower acetic;

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 40 grams, breviscapine 40 grams;

Preparation tablets:

Get danshen root salvianolic acid A and breviscapine, be prepared into 1000 in tablet according to the conventional requirement of tablet pharmaceutics;

The capsule preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into 1000 of capsules according to the conventional requirement of the pharmaceutics of capsule;

The granule preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into 1000 bags of granules according to the conventional requirement of the pharmaceutics of granule;

The soft capsule preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into 1000 of soft capsules according to the conventional requirement of the pharmaceutics of soft capsule;

The pellet preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into pellet 10000 balls according to the conventional requirement of the pharmaceutics of pellet;

The drop pill preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into drop pill 10000 balls according to the conventional requirement of the pharmaceutics of drop pill;

The oral liquid preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into 1000 bottles of oral liquids according to the conventional requirement of the pharmaceutics of oral liquid;

UD is 20mg.

Embodiment 2

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transfers pH value to 7.5,30 ℃ of temperature, heating 1 hour, and solution filters; Filtrating is separated through the HPD-100 macroporous resin column chromatography, and first water, 10% Diluted Alcohol eluting are removed impurity; The ethanol elution of reuse 30% concentration is collected eluent, concentrates; Drying obtains danshen root salvianolic acid A.Danshen root salvianolic acid A content 50.1%.

Or

Radix Salviae Miltiorrhizae extracts with 20% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 9.0,80 ℃ of temperature, heating 6 hours; Solution filters, and filtrating is separated through the HPD-100A macroporous resin column chromatography, first water, 30% Diluted Alcohol eluting; Remove impurity; The ethanol elution of reuse 70% concentration is collected eluent, reclaims ethanol to most; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 20% alcoholic solution eluting discard eluent, and reuse 50% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A.Danshen root salvianolic acid A content 61.3%;

Or

Radix Salviae Miltiorrhizae extracts with 50% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 8.0,50 ℃ of temperature, heating 4 hours; Solution filters, and filtrating is separated through the HPD-300 macroporous resin column chromatography, first water, 20% Diluted Alcohol eluting; Remove impurity; The ethanol elution of reuse 50% concentration is collected eluent, reclaims ethanol to most; Concentrated solution separates with polyamide chromatography post, and first water, 35% alcoholic solution eluting discard eluent, and reuse 75% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 2, through the organic solvent ethyl acetate extraction, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A; Danshen root salvianolic acid A content 90.03%;

Breviscapine is a prior art for preparing, and its content counts 50.01% with lamp-dish flower acetic;

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 40 grams, breviscapine 800 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 840mg.

Embodiment 3

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae extracts with 70% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,120 ℃ of temperature, gauge pressure 0.10MPa pressure, heats 4 hours; Solution filters, and filtrating is separated through the HPD-450 macroporous resin column chromatography, and first water, 20% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 50% concentration is collected eluent, concentrates, and drying obtains danshen root salvianolic acid A, and content is 55.1%.

Or

Radix Salviae Miltiorrhizae extracts with 60% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.0,125 ℃ of temperature, gauge pressure 0.14MPa pressure, heats 2 hours; Solution filters, and filtrating is separated through the D101 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 35% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with polyamide chromatography post, and first water, 25 alcoholic solution eluting discard eluent, and reuse 55% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, and content is 80.1%.

Or

Radix Salviae Miltiorrhizae extracts with 65% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 5.5,130 ℃ of temperature, gauge pressure 0.17MPa pressure, heats 3 hours; Solution filters, and filtrating is separated through the 1300-I macroporous resin column chromatography, and first water, 25% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 45% alcoholic solution eluting discard eluent, and reuse 90% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 4.5, through the organic solvent n-butyl acetate extraction, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, and content is 90.3%

Breviscapine is a prior art for preparing, and its content counts 68.9% with lamp-dish flower acetic; (commercially available)

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 400 grams, breviscapine 40 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 2200mg.

Embodiment 4

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae 35% alcoholic solution extracts and to obtain alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.0,115 ℃ of temperature, gauge pressure 0.07MPa pressure, heats 5 hours; Solution filters, and filtrating is separated through the HPD-100 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 40% concentration is collected eluent, concentrates, and drying obtains danshen root salvianolic acid A, and content is 58.1%.

Or

Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transfers pH value to 4.5,120 ℃ of temperature, gauge pressure 0.10MPa pressure, heats 3.5 hours; Solution filters, and filtrating is separated through the HPD-100A macroporous resin column chromatography, and first water, 20% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 45% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 30% alcoholic solution eluting discard eluent, and reuse 70% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, and content is 69.7%.

Or

Radix Salviae Miltiorrhizae 50% alcoholic solution extracts and to obtain alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,125 ℃ of temperature, gauge pressure 0.14MPa pressure, heats 6 hours; Solution filters, and filtrating is separated through the HPD-300 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 35% alcoholic solution eluting discard eluent, and reuse 60% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 2.5, through the extraction of organic solvent isopropyl alcohol, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, content 91.2%;

Breviscapine is a prior art for preparing, and its content counts 55.2% with lamp-dish flower acetic;

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 200 grams, breviscapine 40 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 2400mg.

Embodiment 5

The preparation of danshen root salvianolic acid A:

Or

Radix Salviae Miltiorrhizae extracts with 60% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.0,130 ℃ of temperature, gauge pressure 0.17MPa pressure, heats 2 hours; Solution filters, and filtrating is separated through the D101 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 35% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with polyamide chromatography post, and first water, 25 alcoholic solution eluting discard eluent, and reuse 55% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, and content is 80.1%.

Or

Radix Salviae Miltiorrhizae extracts with 65% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 5.5,115 ℃ of temperature, gauge pressure 0.08MPa pressure, heats 3 hours; Solution filters, and filtrating is separated through the 1300-I macroporous resin column chromatography, and first water, 25% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 45% alcoholic solution eluting discard eluent, and reuse 90% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 4.5, through the organic solvent n-butyl acetate extraction, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, and content is 90.3%;

Breviscapine is a prior art for preparing, and its content counts 64.3% with lamp-dish flower acetic;

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 40 grams, breviscapine 160 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 200mg.

Embodiment 6

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transfers pH value to 8.5,50 ℃ of temperature, heating 4 hours, heats 3 hours; Solution filters, and filtrating is separated through the 1300-I macroporous resin column chromatography, and first water, 20% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 55% concentration is collected eluent, concentrates, and drying obtains danshen root salvianolic acid A, and content is 52.9%.

Or

Radix Salviae Miltiorrhizae extracts with 50% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 8.0,75 ℃ of temperature, heating 2 hours; Solution filters, and filtrating warp 1400 macroporous resin column chromatographies separate, and first water, 25% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol to most; Concentrated solution separates with polyamide chromatography post, and first water, 25% alcoholic solution eluting discard eluent, and reuse 60% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, and content is 84.1%.

Or

Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transfers pH value to 4.0,110 ℃ of temperature, gauge pressure 0.05MPa pressure, heats 5.5 hours; Solution filters, and filtrating is separated through the AB-8 macroporous resin column chromatography, and first water, 25% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 30% alcoholic solution eluting discard eluent, and reuse 80% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 3.5, through the organic solvent n-butanol extraction, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, and content is 94.7%;

Breviscapine is a prior art for preparing, and its content counts 72.9% with lamp-dish flower acetic;

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 80 grams, breviscapine 120 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 4000mg.

Embodiment 7

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae 35% alcoholic solution extracts and to obtain alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.0,120 ℃ of temperature, gauge pressure 0.10MPa pressure, heats 5 hours; Solution filters, and filtrating is separated through the HPD-100 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 40% concentration is collected eluent, concentrates, and drying obtains danshen root salvianolic acid A, and content is 58.1%.

Or

Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transfers pH value to 4.5,125 ℃ of temperature, gauge pressure 0.14MPa pressure, heats 3.5 hours; Solution filters, and filtrating is separated through the HPD-100A macroporous resin column chromatography, and first water, 20% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 45% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 30% alcoholic solution eluting discard eluent, and reuse 70% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, and content is 69.7%.

Or

Radix Salviae Miltiorrhizae 50% alcoholic solution extracts and to obtain alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,110 ℃ of temperature, gauge pressure 0.05MPa pressure, heats 6 hours; Solution filters, and filtrating is separated through the HPD-300 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 35% alcoholic solution eluting discard eluent, and reuse 60% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 2.5, through the extraction of organic solvent isopropyl alcohol, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, content 91.2%;

Breviscapine is a prior art for preparing, and its content counts 87.3% with lamp-dish flower acetic;

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 360 grams, breviscapine 640 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 1000mg.

Embodiment 8

The preparation of danshen root salvianolic acid A:

Or

Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transfers pH value to 4.0,125 ℃ of temperature, gauge pressure 0.14MPa pressure, heats 5.5 hours; Solution filters, and filtrating is separated through the AB-8 macroporous resin column chromatography, and first water, 25% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 30% alcoholic solution eluting discard eluent, and reuse 80% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 3.5, through the organic solvent n-butanol extraction, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, and content is 94.7%;

Breviscapine is a prior art for preparing, and its content counts 93.8% with lamp-dish flower acetic; (literature method " extracts the technical study of scutellarin ", and " Chinese patent medicine " rolled up 10 phases, 855-856 in 2004 26 from Herba Erigerontis.)

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 40 grams, breviscapine 60 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 50mg.

Embodiment 9

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae extracts with 70% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,120 ℃ of temperature, gauge pressure 0.10MPa pressure, heats 4 hours; Solution filters, and filtrating is separated through the HPD-450 macroporous resin column chromatography, and first water, 20% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 50% concentration is collected eluent, concentrates, and drying obtains danshen root salvianolic acid A, and content is 57.2%.

Or

Radix Salviae Miltiorrhizae extracts with 60% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.0,125 ℃ of temperature, gauge pressure 0.14MPa pressure, heats 2 hours; Solution filters, and filtrating is separated through the D101 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 35% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with polyamide chromatography post, and first water, 25 alcoholic solution eluting discard eluent, and reuse 55% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, and content is 82.3%.

Or

Radix Salviae Miltiorrhizae extracts with 65% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 5.5,130 ℃ of temperature, gauge pressure 0.17MPa pressure, heats 3 hours; Solution filters, and filtrating is separated through the 1300-I macroporous resin column chromatography, and first water, 25% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 45% alcoholic solution eluting discard eluent, and reuse 90% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 4.5, through the organic solvent n-butyl acetate extraction, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, and content is 91.0%;

Breviscapine is a prior art for preparing, and its content counts 69.2% with lamp-dish flower acetic; (commercially available)

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 80 grams, breviscapine 680 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 760mg.

Embodiment 10

The preparation of danshen root salvianolic acid A:

Radix Salviae Miltiorrhizae extracts with 70% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,120 ℃ of temperature, gauge pressure 0.10MPa pressure, heats 4 hours; Solution filters, and filtrating is separated through the HPD-450 macroporous resin column chromatography, and first water, 20% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 50% concentration is collected eluent, concentrates, and drying obtains danshen root salvianolic acid A, and content is 53.8%.

Or

Radix Salviae Miltiorrhizae extracts with 60% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.0,125 ℃ of temperature, gauge pressure 0.14MPa pressure, heats 2 hours; Solution filters, and filtrating is separated through the D101 macroporous resin column chromatography, and first water, 15% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 35% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with polyamide chromatography post, and first water, 25 alcoholic solution eluting discard eluent, and reuse 55% alcoholic solution eluting concentrates, and drying obtains danshen root salvianolic acid A, and content is 79.7%.

Or

Radix Salviae Miltiorrhizae extracts with 65% alcoholic solution and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 5.5,130 ℃ of temperature, gauge pressure 0.17MPa pressure, heats 3 hours; Solution filters, and filtrating is separated through the 1300-I macroporous resin column chromatography, and first water, 25% Diluted Alcohol eluting are removed impurity, and the ethanol elution of reuse 65% concentration is collected eluent, reclaims ethanol extremely to the greatest extent; Concentrated solution separates with the sephadex lh-20 chromatographic column, and first water, 45% alcoholic solution eluting discard eluent, and reuse 90% alcoholic solution eluting reclaims ethanol to most; Concentrated solution is transferred pH value to 4.5, through the organic solvent n-butyl acetate extraction, separates the organic solvent phase, must contain drug solns, concentrates, and drying or lyophilization get danshen root salvianolic acid A, and content is 91.2%;

Breviscapine is a prior art for preparing, and its content counts 70.1% with lamp-dish flower acetic; (commercially available)

Formulation preparation:

Crude drug is: danshen root salvianolic acid A 40 grams, breviscapine 720 grams;

Preparation tablets:

The capsule preparation:

The granule preparation:

The soft capsule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

UD is 760mg.

Annotate: the present invention's concrete technical scheme required for protection is not limited to the concrete combination of the expressed technical scheme of the foregoing description.

Claims

1. method for preparing that contains the combination of oral medication of danshen root salvianolic acid A; Wherein pharmaceutical composition is a danshen root salvianolic acid A 1-10 weight portion; Breviscapine 1-20 weight portion; Wherein preparation of pharmaceutical compositions becomes tablet, capsule, granule, pellet, drop pill, the oral liquid of UD, and wherein the crude drug of tablet, capsule, granule, pellet, drop pill, oral liquid comprises breviscapine, it is characterized by:

Danshen root salvianolic acid A extracts purification:

Or:

Formulation preparation:

Preparation tablets:

The capsule preparation:

The granule preparation:

The pellet preparation:

The drop pill preparation:

The oral liquid preparation:

Get danshen root salvianolic acid A and breviscapine, be prepared into oral liquid according to the conventional requirement of the pharmaceutics of oral liquid.