CN101019824A - Oral medicine composition containing salvianolic acid and its prepn - Google Patents
Oral medicine composition containing salvianolic acid and its prepn Download PDFInfo
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- CN101019824A CN101019824A CN 200710090922 CN200710090922A CN101019824A CN 101019824 A CN101019824 A CN 101019824A CN 200710090922 CN200710090922 CN 200710090922 CN 200710090922 A CN200710090922 A CN 200710090922A CN 101019824 A CN101019824 A CN 101019824A
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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
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, have only about 5/10000ths, even it is extracted by a series of technology, 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 with the more weak composition of other relativity of Radix Salviae Miltiorrhizae (such as TANSHINONES, salvianolic acid B etc.) compatibility is 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, particularly the compatibility of the salvianolic acid A of level and pharmaceutically active ingredient in other, one of difficult problem that need be to be solved especially in batches.
Breviscapine is the flavonoids effective constituent that extracts from Compositae Herba Erigerontis aceris plant Herba Erigerontis (Erigeron brieviscapus (Vant) Hand Mazz.), 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., these preparations have certain therapeutical effect in clinical practice to the thrombosis cardiovascular and cerebrovascular disease, but in the time of the molten diffusing recovery organization organ blood flow of thrombosis, generation reperfusion injury that will be in various degree, can cause even more serious consequence, 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, pour into the damage that produces again after reducing thrombolytic, these simple clinically compatibilities are though can reach certain therapeutical effect, but can produce a large amount of untoward reaction, bring huge medication difficulty to the doctor, bring bigger misery to the patient, therefore, should be at the compatibility of middle pharmaceutically active ingredient or component effective site, carry out a series of experiment, the combination of pharmaceutically active ingredient or component effective site in the excavation is 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, but there is dispute in its science always, therefore middle pharmaceutically active ingredient or component effective site are carried out compatibility, admitted by numerous researchers, 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 using, quantitative addition, the counteracting of toxic reaction that neither be mechanical, but by a series of research (medicine efficacy screening, aspects such as 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, but because prior art exist certain defective: obtain salvia miltiorrhiza tanshinoate and be salvianolic acid class material based on 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 determine.
Summary of the invention
For these reasons, we are by carrying out a series of Study on Transformation to red rooted salvia, be about to Radix Salviae Miltiorrhizae total phenolic acids and obtained the danshen root salvianolic acid A of level in batches by certain chemical reaction, mode by oral administration again, with medicine efficacy screening, system's drug effect and safety evaluatio, experiments such as pharmacokinetics have determined that a certain amount of danshen root salvianolic acid A and a certain amount of breviscapine can make up, this drug regimen is except having complementary effect, can also well solve the reperfusion injury that produces behind the thrombolytic, 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 by 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 unit dose;
Wherein unit dose is 20mg-4000mg; Wherein preferred unit dosage is 50-2000mg.Unit dose is lower than 20mg does not have effect in clinical use, unit dose 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 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, the accent pH value is to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure, heats 1-6 hour; Solution filters, filtrate is separated through nonpolar or low pole macroporous resin column chromatography, 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, the accent pH value is to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure, heats 1-6 hour; Solution filters, filtrate is separated through nonpolar or low pole macroporous resin column chromatography, 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, 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, the accent pH value is to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure, heats 1-6 hour; Solution filters, filtrate is separated through nonpolar or low pole macroporous resin column chromatography, 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, 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 reduces to 80% by 10% ratio that rises to 20%, 0.2% aqueous acetic acid by 90%; In the time of 15-55 minute, the ratio of acetonitrile reduces to 70% by 20% ratio that rises to 30%, 0.2% aqueous acetic acid by 80%; In the time of 55-65 minute, the ratio of acetonitrile reduces to 50% by 30% ratio that rises to 50%, 0.2% aqueous acetic acid by 70%; In the time of 65-72 minute, the ratio of acetonitrile reduces to 20% by 50% ratio that rises to 80%, 0.2% aqueous acetic acid 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 rises to 90% by 80% ratio of reducing to 10%, 0.2% aqueous acetic acid 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;
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;
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;
Experimental result sees 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 is a raw material) mg/ sheet the application capsule (salvianolic acid A, breviscapine is a raw material) mg/ grain the application granule (salvianolic acid A, breviscapine is a raw material) mg/ bag the application soft capsule (salvianolic acid A, breviscapine is a raw material) mg/ grain the application pellet (salvianolic acid A, breviscapine is a raw material) mg/10 ball the application drop pill agent (salvianolic acid A, breviscapine is a raw material) the 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 by 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 the pre-raising of equivalent environment 2 days, free diet.After pre-raising finishes, 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.Adjust respiration parameter according to the respiratory frequency and the degree of depth.Connect electrocardiograph subsequently, survey normal ECG.Cut off front field of operation hair, iodine disinfection, cut off skin, subcutaneous tissue, front muscle and fascia 3～4cm, it is long to separate Intercostal muscle 3cm with the 18# vascular forceps along the 3rd intercostal passivity, open thoracic cavity and pericardium, recording ecg, strut 3,4 ribs refer to hold thoracic cavity, rat right side with left hand four, and the assistant upwards pushes away thymus with the ophthalmology tweezer, between left auricle and pulmonary conus, find ligation sign blood vessel great cardiac vein, 2mm place noinvasive roundlet pin band 6-0 silk thread threading below left auricle, 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 thereon 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.
| Infarction size (%)= | The necrotic area cardiac weight | ×100％ |
| Necrotic area+non-necrotic area cardiac weight |
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 (%) |
| Control 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 adjusting grouping, 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 distilled water melt into 100ml, afternoon 1, group dosage gastric infusion was pressed in point-2, 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 claims body weight weekly one time, adjusts dosage by body weight, and measures food ration.4 weeks of administration are detected serum TC, TG, LDL-C and HDL-C level respectively, and experimental result sees Table 4
Table 4 administration 4 all red lamps 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 sees Table 5:
Table 5 pair MACO rat cerebral tissue water content
| Group | Left hemisphere brain water content % | Right hemisphere brain water content % |
| Control group Nimodipine prescription case 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8 schemes 9 | 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 77.97±0.46 | 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 * 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 3: danshen root salvianolic acid A 8 weight portions, breviscapine 6 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, make 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, adjusts PH,, be prepared into aseptic urate crystal through stirring, filtration, cold drying, screening, sterilization with HCl.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+ concentration thereof, experimental result sees 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 |
| 3 groups of normal group gout model group scheme 1 prescription case 2 prescription cases | 10 10 10 10 10 | 0.89±0.07 1.28±0.11 1.10±0.09 ** 1.07±0.08 ** 1.08±0.08 ** |
Annotate: compare with the gout model group
*P＜0.01
Pharmacological evaluation brief summary: show that by 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 4: danshen root salvianolic acid A 1 weight portion, breviscapine 5 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 7: danshen root salvianolic acid A 5 weight portions, breviscapine 1 weight portion;
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 by 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 sees 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 schemes 19 | 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 - 0.41 |
Five. toxicologic study
Experimental program:
Scheme 1: danshen root salvianolic acid A 1 weight portion, breviscapine 21 weight portions;
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 sees 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 schemes 11 | 17.1 20.3 20.7 19.8 19.7 18.4 19.2 20.1 15.8 15.4 12.91 |
Experiment conclusion: by pharmacodynamics experiments, pharmacokinetics experiment, toxicological experiment, we determine 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, solution filters, filtrate 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, collect eluent, concentrate drying, obtain 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 filtrate 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 to most; 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 filtrate 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 to most; 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;
Unit dose is 20mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
Embodiment 2
The preparation of danshen root salvianolic acid A:
Radix Salviae Miltiorrhizae obtains aqueous extract with water extraction, transferred pH value to 7.5,30 ℃ of temperature, heating 1 hour, solution filters, and filtrate is separated through the HPD-100 macroporous resin column chromatography, first water, 10% Diluted Alcohol eluting, remove 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, alcohol extract concentrates ethanol to most, transferred pH value to 9.0,80 ℃ of temperature, heating 6 hours, solution filters, and filtrate 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, alcohol extract concentrates ethanol to most, transferred pH value to 8.0,50 ℃ of temperature, heating 4 hours, solution filters, and filtrate 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:
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;
Unit dose is 840mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
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 filtrate 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 filtrate 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 to most; 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 filtrate 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 to most; 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:
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;
Unit dose is 2200mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
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 filtrate 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 filtrate 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 to most; 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 filtrate 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 to most; 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:
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;
Unit dose is 2400mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
Embodiment 5
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 filtrate 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 4.0,130 ℃ of temperature, gauge pressure 0.17MPa pressure, heats 2 hours; Solution filters, and filtrate 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 to most; 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 filtrate 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 to most; 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:
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;
Unit dose is 200mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
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 filtrate 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 filtrate is separated through 1400 macroporous resin column chromatographies, 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 filtrate 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 to most; 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:
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;
Unit dose is 4000mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
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 filtrate 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 filtrate 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 to most; 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 filtrate 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 to most; 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:
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;
Unit dose is 1000mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
Embodiment 8
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 filtrate 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 filtrate is separated through 1400 macroporous resin column chromatographies, 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 4.0,125 ℃ of temperature, gauge pressure 0.14MPa pressure, heats 5.5 hours; Solution filters, and filtrate 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 to most; 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:
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;
Unit dose is 50mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
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 filtrate 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 filtrate 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 to most; 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 filtrate 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 to most; 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:
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;
Unit dose is 760mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
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 filtrate 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 filtrate 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 to most; 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 filtrate 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 to most; 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:
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;
Unit dose is 760mg.
The combination of oral medication that contains danshen root salvianolic acid A treats and/or prevents application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine in preparation.
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 (10)
1. a combination of oral medication that contains danshen root salvianolic acid A is characterized in that: danshen root salvianolic acid A 1-10 weight portion, breviscapine 1-20 weight portion.
2. a kind of combination of oral medication that contains danshen root salvianolic acid A according to claim 1, danshen root salvianolic acid A 1-5 weight portion wherein, breviscapine 1-5 weight portion.
3. a kind of combination of oral medication that contains danshen root salvianolic acid A according to claim 1 and 2, wherein breviscapine content in lamp-dish flower acetic more than or equal to 50% and less than 100%.
4. a kind of combination of oral medication that contains danshen root salvianolic acid A according to claim 1 and 2, wherein the content of danshen root salvianolic acid A is more than or equal to 50% and less than 100%.
5. a kind of combination of oral medication that contains danshen root salvianolic acid A according to claim 1 and 2, wherein preparation of pharmaceutical compositions becomes tablet, capsule, granule, soft capsule, pellet, drop pill, the oral liquid of unit dose.
6. a kind of combination of oral medication that contains danshen root salvianolic acid A according to claim 5, wherein unit dose is 20mg-4000mg.
7. a kind of combination of oral medication that contains danshen root salvianolic acid A according to claim 5, wherein unit dose is 50-2000mg.
8. a kind of combination of oral medication that contains danshen root salvianolic acid A according to claim 5, wherein the crude drug of tablet, capsule, granule, soft capsule, pellet, drop pill, oral liquid comprises breviscapine, its preparation method is:
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, the accent pH value is to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure, heats 1-6 hour; Solution filters, filtrate is separated through nonpolar or low pole macroporous resin column chromatography, 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, the accent pH value is to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure, heats 1-6 hour; Solution filters, filtrate is separated through nonpolar or low pole macroporous resin column chromatography, 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, 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, the accent pH value is to 7.5-9.0,30-80 ℃ temperature, heating 1-6 hour or transfer pH value to 3.5-6.0,110-130 ℃ temperature, gauge pressure 0.05MPa-0.17MPa pressure, heats 1-6 hour; Solution filters, filtrate is separated through nonpolar or low pole macroporous resin column chromatography, 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, 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;
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
9. according to claim 1,2,8 each described a kind of combination of oral medication that contain danshen root salvianolic acid A, it is characterized in that 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; Number of theoretical plate should be not less than 60000 by salvianolic acid A; 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 reduces to 80% by 10% ratio that rises to 20%, 0.2% aqueous acetic acid by 90%; In the time of 15-55 minute, the ratio of acetonitrile reduces to 70% by 20% ratio that rises to 30%, 0.2% aqueous acetic acid by 80%; In the time of 55-65 minute, the ratio of acetonitrile reduces to 50% by 30% ratio that rises to 50%, 0.2% aqueous acetic acid by 70%; In the time of 65-72 minute, the ratio of acetonitrile reduces to 20% by 50% ratio that rises to 80%, 0.2% aqueous acetic acid 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 rises to 90% by 80% ratio of reducing to 10%, 0.2% aqueous acetic acid 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;
(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;
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.
10. treat and/or prevent application in cardiovascular and cerebrovascular disease, hyperlipidemia, gout, hepatic injury, hepatic fibrosis, pulmonary fibrosis, tumor, the old and feeble medicine according to claim 1,2 each described a kind of combination of oral medication that contain danshen root salvianolic acid A in preparation.
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| CN2007100909226A CN101019824B (en) | 2007-03-27 | 2007-03-27 | Preparation method of oral medicine composition containing salvianolic acid |
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| CN2007100909226A CN101019824B (en) | 2007-03-27 | 2007-03-27 | Preparation method of oral medicine composition containing salvianolic acid |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102423310A (en) * | 2012-01-04 | 2012-04-25 | 中国药科大学 | Application of salvianolic acid C in preparation of drugs for prevention and treatment of hyperuricemia |
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| CN107137410B (en) * | 2017-06-08 | 2019-11-26 | 中国人民解放军第四军医大学 | It is a kind of for treating the Compound Chinese Herbal Monomer Recipe compatibility agent and preparation method of cerebral ischemia |
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| CN102423310A (en) * | 2012-01-04 | 2012-04-25 | 中国药科大学 | Application of salvianolic acid C in preparation of drugs for prevention and treatment of hyperuricemia |
| CN102423310B (en) * | 2012-01-04 | 2012-12-12 | 中国药科大学 | Application of salvianolic acid C in preparation of drugs for prevention and treatment of hyperuricemia |
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