CN1299717C

CN1299717C - Pharmaceutical composition for treating cerebral apoplexy and its preparation method

Info

Publication number: CN1299717C
Application number: CNB2004100580111A
Authority: CN
Inventors: 王永炎; 李澎涛; 潘彦舒; 徐雅; 谢利军
Original assignee: Beijing University of Chinese Medicine
Current assignee: Beijing University of Chinese Medicine
Priority date: 2004-08-06
Filing date: 2004-08-06
Publication date: 2007-02-14
Anticipated expiration: 2024-08-06
Also published as: CN1602893A

Abstract

本发明公开了一种药物组合物及其制备方法，该组合物由牛胆酸、异去氧胆酸、黄芩苷、栀子、珍珠母制成，先制备栀子提取液和珍珠母粉水解液，再加异去氧胆酸粉和牛胆酸粉调pH得混合药液；将混合药液经醇沉后，加入黄芩苷细粉，经调pH等工艺，得总混合药液；再取总混合药液制备成各种制剂。本发明改进了栀子提取工艺，形成以栀子苷为主要组分的半成品。在保持清开灵注射液治疗脑栓塞类疾病生物效应的前提下，基本消除原存在的不良反应，达到了稳效减毒的目的，可用于治疗脑栓塞、脑出血类疾病的新药开发。The invention discloses a pharmaceutical composition and a preparation method thereof. The composition is prepared from taurocholic acid, isodeoxycholic acid, baicalin, gardenia and mother-of-pearl. The gardenia extract and mother-of-pearl powder are hydrolyzed first liquid, add isodeoxycholic acid powder and taurocholic acid powder to adjust the pH to obtain a mixed medicinal liquid; after the mixed medicinal liquid is ethanol-precipitated, add baicalin fine powder, and adjust the pH and other processes to obtain a total mixed medicinal liquid; The total mixed medicinal solution is prepared into various preparations. The invention improves the gardenia extraction process and forms a semi-finished product with geniposide as the main component. On the premise of maintaining the biological effect of Qingkailing injection in the treatment of cerebral embolism diseases, the existing adverse reactions are basically eliminated, and the goal of stable effect and attenuation is achieved. It can be used for the development of new drugs for the treatment of cerebral embolism and cerebral hemorrhage.

Description

A kind of pharmaceutical composition for the treatment of apoplexy and preparation method thereof

Technical field

The present invention relates to a kind of pharmaceutical composition and preparation method thereof, particularly relate to a kind of pharmaceutical composition for the treatment of apoplexy and preparation method thereof, belong to medical technical field.

Background technology

QINKAILING ZHUSHEYE is the compound injection of a kind of Chinese medicine that classical prescription cow-bezoar bolus for resurrection comes through science development purification.It has the effect of heat-clearing and toxic substances removing, eleminating phlegm and freeing channels, consciousness and resuscitation restoring, is mainly used in diseases such as hyperpyrexia, stupor, apoplectic hemiplegia, acute, chronic hepatitis, upper respiratory tract infection, pneumonia clinically.Clinical practice is over 22 years because its curative effect is obvious, uses increasing, since nineteen ninety-two continuous three times by authorization for national hospital of traditional Chinese hospital emergency treatment first-selection required medicines, be listed as " national Chinese medicine protection kind ".Over particularly past 10 years, the more clinical extensive approval of its unique curative effect aspect cerebrovascular disease.

But in recent years, the QINKAILING ZHUSHEYE untoward reaction takes place often, and relevant report also day by day increases, even some serious adverse effects occurred, causes death.Main adverse reaction comprises: allergy (erythra, pruritus, exfoliative dermatitis, vasodilation, anaphylactic shock etc.), central nervous system's untoward reaction (disturbance of consciousness, stupor, tic, mental disorder etc.) and accidental feel sick, vomiting, stomachache, phlebitis, acute renal failure etc.Therefore, improve production technology, improve pharmaceutical purity, the impurity content of no pharmacological action is reduced to minimum, reducing untoward reaction has been the task of top priority.

Because QINKAILING ZHUSHEYE is a Chinese medicine compound injection, the effect scope is wide, and the sick kind of treatment is many, and is unclear again to the effective substance of every Herba indigoferae Pseudotinctoriae, the process modification difficulty.Therefore, to plant the research carry out QINKAILING ZHUSHEYE prescription and technology be feasible way at curing mainly disease.

Summary of the invention

The object of the present invention is to provide a kind of pharmaceutical composition; The present invention also aims to provide a kind of pharmaceutical composition for the treatment of apoplexy and preparation method thereof.

The objective of the invention is to be achieved through the following technical solutions:

Pharmaceutical composition of the present invention is:

Cholic acid 3-9 weight portion, baicalin 4-6 weight portion, Fructus Gardeniae 20-35 weight portion, Concha Margaritifera powder 40-60 weight portion; The preferable range of aforementioned pharmaceutical compositions is: cholic acid 6.5-7.4 weight portion, baicalin 5.0-5.5 weight portion, Fructus Gardeniae 25.0-28.0 weight portion, Concha Margaritifera powder 45.0-50.0 weight portion;

The optimum ratio of aforementioned pharmaceutical compositions is: cholic acid 4 weight portions, baicalin 4.5 weight portions, Fructus Gardeniae 23 weight portions, Concha Margaritifera powder 42 weight portions; Cholic acid 8 weight portions, baicalin 5.6 weight portions, Fructus Gardeniae 30 weight portions, Concha Margaritifera powder 55 weight portions; Cholic acid 7.4 weight portions, baicalin 5.0 weight portions, Fructus Gardeniae 28 weight portions, Concha Margaritifera powder 45 weight portions; Cholic acid 6.5 weight portions, baicalin 5.5 weight portions, Fructus Gardeniae 25.0 weight portions, Concha Margaritifera powder 50.0 weight portions; Or cholic acid 7 weight portions, baicalin 5.3 weight portions, Fructus Gardeniae 26 weight portions, Concha Margaritifera powder 48 weight portions.

In the aforementioned pharmaceutical compositions, described cholic acid is made up of in the ratio of 2-4: 2-4.5 taurine and hyodesoxycholic acid; Wherein the preferred proportion scope of taurine and hyodesoxycholic acid is: 3.0-3.5: 3.5-3.9; The optimum ratio of taurine and hyodesoxycholic acid is: 2.2: 3.4, and 3.9: 4.1 or 3.2: 3.8.

In the aforementioned pharmaceutical compositions, described baicalin can use noroxylin, nor-baicalin element, wogonoside element or wogonoside to replace.

Get aforementioned pharmaceutical compositions, press the pharmaceutics conventional method, be prepared into various clinical acceptable forms, include but not limited to a kind of in the middle of the following dosage form: ejection preparation, tablet, capsule, spray, pill, granule, suspensoid, drop pill, oral liquid etc.; Described ejection preparation can be injection, infusion solutions, freeze-dried powder, injection tablet, suspension type injection etc.

Preparation of drug combination technology of the present invention comprises the steps:

A. measure taurine, hyodesoxycholic acid, baicalin, Fructus Gardeniae, Concha Margaritifera powder by prescription;

B. prepare gardenia extract and Concha Margaritifera powder hydrolyzed solution, mix homogeneously is transferred PH to 7, gets mixed extract;

C. get ethanol, transfer PH to 11, add the deoxycholic acid powder and make whole dissolvings, add the taurine powder and transfer PH to 9 to make it whole dissolvings; Mixed extract is mixed in the cholic acid solution, gets admixing medical solutions; With the admixing medical solutions ethanol precipitation, cold preservation, sucking filtration reclaims ethanol; Add the injection water, add the baicalin fine powder, it is dissolved fully, get total admixing medical solutions;

D. get total admixing medical solutions, add adjuvant, preparation process or step are made clinical acceptable forms: ejection preparation, tablet, capsule, spray, pill, granule, suspensoid, drop pill, oral liquid etc. routinely.

Aforesaid preparation of drug combination method of the present invention, wherein gardenia extract can be prepared by following method:

Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, according to the percolation under the fluid extract item (" appendix IO of Chinese pharmacopoeia version in 2000),, collect percolate with 60%～80% ethanol percolation of 6～10 times of amounts, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear paste, add 3～5 times of water gagings dilutions, stir evenly, 0-4 ℃ of cold preservation 36～60 hours, filter, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, it is colourless earlier to be eluted to eluent with distilled water, 8～12 times of amounts of reuse, 60%～80% ethanol elution is collected eluent, and decompression recycling ethanol also concentrates, drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with water for injection; Regulate pH value to 6.0～7.0,0-4 ℃ of cold preservation 12～36 hours, 0.45 μ m filter membrane filters, and filtrate adds the injection water to dissolving, filters with 0.22 μ m filter membrane, gets gardenia extract.

Aforesaid preparation of drug combination method of the present invention, wherein the Concha Margaritifera powder hydrolyzed solution can be prepared by following method:

8 times of H that measure 4N of preparation earlier ₂SO ₄Aqueous solution; Add Concha Margaritifera powder, being heated to boils kept little 6-7 of boiling hour, filtered, and filtering residue gets the clear and bright solution of salmon pink with hot wash 2 times; Cooling back adularescent acicular crystals is separated out, and refilters, and removes crystal, gets the salmon pink settled solution; Cooling when clarifying hydrolyzed solution is concentrated an only surplus 2-3 who is equivalent to the raw material total amount and doubly measures adds ethanol and transfers to and contain the alcohol amount and reach 60%, cold preservation 24 hours; Sucking filtration, filtrate recycling ethanol takes out cold preservation and uses for dosing to there not being the alcohol flavor;

Aforesaid preparation of drug combination method of the present invention, wherein said dosing step can be specific as follows:

1, extracts the mixing of medicinal liquid

Gardenia extract, Concha Margaritifera powder hydrolyzed solution are filtered respectively, and mix homogeneously is transferred PH to 7 with 10%NaOH liquid, and sucking filtration gets mixed extract;

2, the dissolving of cholic acid

Be taken as 60 times 75% ethanol of cholic acid amount, transfer PH to 11, add the deoxycholic acid powder and make whole dissolvings, add the taurine powder and transfer PH to 9 to make it whole dissolvings with 10%NaOH liquid; Mixed extract is mixed in the cholic acid solution, gets admixing medical solutions;

3, the alcohol of admixing medical solutions is handled

Admixing medical solutions is added 95% ethanol and is made it contain alcohol amount to reach 75%; Transfer PH to 7,2-3 days sucking filtration of sealing cold preservation with 10%NaOH liquid, reclaim ethanol to there not being the alcohol flavor, room temperature is placed; Add fresh water for injection, transfer to neutral 4/5 o'clock to full dose, transfer PH to 7.2 with 10%NaOH liquid, cold preservation 3-4 days, sucking filtration got supernatant liquid;

4, the processing of baicalin dissolving and mixing

Get the fresh deionized water of total approximately dosing volume, transfer PH to 8, add the baicalin fine powder, transfer PH to 7, it is dissolved fully with 10%NaOH solution with 10%NaOH solution; To contain baicalin solution and join in the admixing medical solutions, get total admixing medical solutions.

The preparation technology of pharmaceutical composition ejection preparation of the present invention comprises the steps:

Make total admixing medical solutions by above-mentioned preparation method, transfer PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, with G4 filter bulb sucking filtration, makes ejection preparation.

The preparation technology of medicament composition dropping pills preparation of the present invention comprises the steps:

Make total admixing medical solutions by above-mentioned preparation method, transfer PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, with G4 filter bulb sucking filtration, concentrating under reduced pressure, spraying drying powder-forming; (ratio is PEG4000: stearic acid: paraffin: medicated powder=3.60: 0.32: 0.08: 1.00) water-bath is dissolved with this medicated powder and stroma ground substance, mix homogeneously, with dropper under 75 ℃, splash into from top to bottom in the liquid paraffin liquid coolant, collect drop pill and absorb coolant, promptly get medicament composition dropping pills of the present invention with filter paper.

The active component of the present composition comprises cholic acid class (taurine, hyocholic acid and hyodesoxycholic acid etc.), scutellaria glycosides (baicalin is main, also has wogonoside, scutellarin etc.), capejasmine cycloolefines ether terpene class (jasminoidin is master, geniposide and gardenia acid etc.) and pigment (crocin, crocetin etc.), combined state calcium and free state calcium etc.

With the prepared injection of the described preparation method of technical solution of the present invention, to compare with obtained injection after the method that changes the Concha Margaritifera hydrolysis with QINKAILING ZHUSHEYE, color is the most shallow, and its total solid matters amount is less.Adopt Waster rat MCAT cerebral infarction model, and be contrast, measure cerebral tissue SOD, MDA, NOS content etc. with the QINKAILING ZHUSHEYE; The result shows that at the treatment of cerebral ischemia, the present composition is approaching substantially with QINGKAILING on some indexs, and is particularly evident by the injection effect that the described method of technical scheme makes, and also is better than QINGKAILING aspect some under some proportionings.

According to related experiment, the result shows: new pharmaceutical composition of the present invention and preparation technology thereof, not only dispeled the flavour of a drug that may cause untoward reaction, and reduced untoward reaction, reduce to the impurity content of no pharmacological action minimum; Medicine reduces to four kinds (wherein taurine, hyodesoxycholic acid are collectively referred to as cholic acid), has improved the precision of Fructus Gardeniae extract; And its biological effect and former QINKAILING ZHUSHEYE are to the difference of the protective effect not statistically signigicant of cerebral ischemia; the basic cerebral protection of QINKAILING ZHUSHEYE in the cerebral ischemia pathological process that keep; dispel the composition that may cause untoward reaction simultaneously, reached the optimization purpose of steady effect attenuation.Pharmaceutical composition of the present invention and QINGKAILING relatively have following advantage on pharmacodynamics: pharmaceutical composition 1. of the present invention is suppressing the microvessel cell adhesion process that cerebral ischemia starts, thereby improve ischemic tissue's microcirculation perfusion, improve the hypoxic-ischemic state aspect and have more significant advantage; 2. pharmaceutical composition of the present invention is alleviating ischemia to more remarkable aspect neuron and the injury of vascular endothelial cells.

Following experimental example is used to further specify but is not limited to the present invention.

Experimental example 1 cerebral ischemia animal model is to the research of QINGKAILING effective substance and compatibility effect feature

Utilize the Focal Cerebral Ischemia Reperfusion model of line bolt method intraluminal middle cerebral artery occlusion in rats infraction, folder closes the incomplete cerebral ischemia animal model of rat bilateral common carotid arteries repeatedly, adopt the neurologic impairment and the mortality rate overall evaluation, in conjunction with the biochemical analysis of serial metabolic damage criterion, carried out the screening analysis of active component in the QINKAILING ZHUSHEYE effective substance.

A. group effect: comprise QINGKAILING each several part effective substances such as cholic acid, baicalin, Fructus Gardeniae extract, Flos Lonicerae extract, Radix Isatidis extract, Concha Margaritifera and Cornu Bubali hydrolysate, all can significantly improve the neurologic impairment symptom of animal pattern.Demonstrate significant synergism between each several part, the full side's effect of QINGKAILING is the most remarkable.

Cholic acid, baicalin can reduce the animal pattern mortality rate, and all the other single parts are not to the influence of animal pattern mortality rate significantly or the trend of increasing the weight of arranged.Each compatibility group mortality rate significantly descends, and along with increasing of each several part effective substance compatibility, mortality rate further descends, and the mortality rate of the full side of QINGKAILING is zero, has shown compatibility synergism and attenuation preferably.

B. anti peroxidation of lipid reaction: Flos Lonicerae, Radix Isatidis, Concha Margaritifera and Cornu Bubali can significantly improve the SOD level of animal pattern cerebral tissue, and cholic acid, baicalin, Fructus Gardeniae do not have the improvement effect to cerebral tissue SOD level.Baicalin and Fructus Gardeniae compatibility have produced the effect of significant raising animal pattern cerebral tissue SOD level, and all the other compatibilities do not show synergistic function.Radix Isatidis can significantly reduce the LPO content of animal pattern cerebral tissue, and all the other each medicines all do not make significant difference to this.But can reduce cerebral tissue LPO content significantly behind baicalin and the Fructus Gardeniae compatibility, cerebral tissue LPO content has also obtained effectively reducing behind cholic acid and Concha Margaritifera, the Cornu Bubali compatibility, and Fructus Gardeniae, Radix Isatidis and cholic acid and Concha Margaritifera, Cornu Bubali compatibility all can significantly reduce the LPO content of cerebral tissue.And baicalin, Flos Lonicerae do not make significant difference to this lipid peroxidation process.

C. to the influence of brain energy metabolism: Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali can significantly reduce the activity of sodium-potassium in the animal pattern cerebral tissue-ATP enzyme, and the effect of baicalin is opposite, and cholic acid, Flos Lonicerae do not have remarkable effect.With cholic acid and Concha Margaritifera, Cornu Bubali compatibility and with baicalin and Fructus Gardeniae compatibility, all can weaken the reduction effect of the latter to cerebral tissue sodium-potassium-atpase activity, point out 5 pairs of sodium-potassium-atpase activities of this two assembly all may exist antagonism.Flos Lonicerae and Radix Isatidis compatibility do not present the obvious synergistic effect.

Concha Margaritifera and Cornu Bubali have significant improvement trend to the brain-capacity lotus, and cholic acid, baicalin, Fructus Gardeniae do not have remarkable effect, and Flos Lonicerae, Radix Isatidis have certain improvement trend.Still kept original effect behind Concha Margaritifera, Cornu Bubali and the cholic acid compatibility, and behind baicalin and Fructus Gardeniae, Flos Lonicerae and the Radix Isatidis compatibility variation of brain-capacity lotus is not still made significant difference, Concha Margaritifera, Cornu Bubali and baicalin, Fructus Gardeniae, Flos Lonicerae and Radix Isatidis have then produced negative influence to its original effect.

D. the property damaged indicator reaction analysis: Radix Isatidis can significantly reduce the level of vWF in the animal pattern blood plasma, and cholic acid, baicalin, Fructus Gardeniae, Flos Lonicerae, Concha Margaritifera and Cornu Bubali do not have remarkable improvement effect to animal pattern vWF level, cholic acid even can produce passive influence.But, cholic acid and Concha Margaritifera, Cornu Bubali compatibility, and baicalin and Fructus Gardeniae compatibility, all produced the effect of significant reduction animal pattern plasma vWF level, two assembly 5 have synergism, Flos Lonicerae cooperates with Radix Isatidis and does not then show significant synergism, and the effect of the full side of QINGKAILING is more remarkable.Cholic acid, baicalin can significantly reduce the NOS level of animal pattern cerebral tissue, and Fructus Gardeniae, Flos Lonicerae, Radix Isatidis, Concha Margaritifera and Cornu Bubali do not have the improvement effect to animal pattern cerebral tissue NOS level.Compatibility effect does not show the obvious synergistic effect.

Above-mentioned result of study shows: each ingredient of QINKAILING ZHUSHEYE has relative specificity for the influence of cerebral ischemia different pathological link, exists simultaneously effectively and many-sided effect relations such as invalid, compatibility Synergistic and attenuation.

Experimental example 2 application cell models are to the screening study of QINGKAILING effective substance

Use neuron, neurogliocyte and the endotheliocyte cultivated, under the nontoxic dosage of determining normal cultured cell, carried out the effective substance screening of cell model.

A. to neuronic influence: there is significant protective effect the full side of QINGKAILING to neuron when doses.The mixed liquid of Fructus Gardeniae wherein, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate compatibility has significant protective effect to neuron, promote the propagation of neuron (SH-SY5Y), suppress apoptosis in neuronal, suppress the generation of lipid peroxide, increase the SOD and the NOS vigor of anti peroxidation of lipid system system, and improve NO concentration; The mixed liquid of taurine, hyocholic acid, Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate compatibility, neuron has been produced tangible toxic action, it suppresses apoptosis in neuronal, suppress the generation of lipid peroxide, increase scavenger SOD and NOS vigor, and the effect that improves NO concentration, decrease than the mixed liquid of Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate compatibility.Studies show that of single medicine: during doses; taurine, Fructus Gardeniae have significant protective effect to neuron; and hyocholic acid, baicalin have higher toxic action to neuron; but the baicalin of high dose is light than low dose group to the neuronic inhibitory action of SH-SY5Y, and the Flos Lonicerae of low dosage, Concha Margaritifera and Cornu Bubali hydrolysate and heavy dose of Radix Isatidis have significant protective effect to neuron.So the compatibility of various dose, the QINGKAILING effective substance of different component compatibility can reduce the deficiency of each single medicinal material, and reach the optimum medicine efficacy of QINGKAILING.

B. to the influence of endotheliocyte: there is significant protective effect the full side of QINGKAILING to endotheliocyte when doses.Wherein the mixed liquid of Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate compatibility has significant protective effect to endotheliocyte, promotes the propagation of endotheliocyte (ECV304); The mixed liquid of taurine, hyocholic acid, Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate compatibility promotes the effect of endothelial cell proliferation, is weaker than the compatibility effect of Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate, and toxic action increases.Studies show that of one-component: taurine, baicalin, Concha Margaritifera and Cornu Bubali hydrolysate have the effect of obvious promotion endothelial cell proliferation, heavy dose of Fructus Gardeniae, hyocholic acid also has the effect of stronger promotion endothelial cell proliferation, and Flos Lonicerae and Radix Isatidis have the obvious suppression effect to endothelial cell growth.

C. to the influence of neurogliocyte: the full side of QINGKAILING has than the obvious suppression effect neurogliocyte, and the full liquid of the QINGKAILING of low dosage significantly is lower than heavy dose to the inhibitory action of neurogliocyte.Wherein, the mixed liquid of Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate compatibility has remarkable protective effect to neurogliocyte.The mixed liquid of taurine, hyocholic acid, Fructus Gardeniae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate compatibility has significant inhibitory effect to neurogliocyte.Low dosage taurine, hyocholic acid, baicalin have remarkable protective effect to neurogliocyte, and high dose taurine, hyocholic acid, baicalin have higher inhibitory action to neurogliocyte.Fructus Gardeniae, Flos Lonicerae, Radix Isatidis, Concha Margaritifera and Cornu Bubali hydrolysate all have slight inhibitory action to neurogliocyte.

In view of QINKAILING ZHUSHEYE clinical adverse feature, at first in line with protecting the principle of imitating attenuation, determine that the QINGKAILING untoward reaction and the part of no remarkable effect are Cornu Bubali, Flos Lonicerae and Radix Isatidis, effective prescription of treatment cerebral ischemia is a pharmaceutical composition of the present invention.

The preferred research of experimental example 3 pharmaceutical composition compatibility quantitative changeizations of the present invention

Pharmaceutical composition of the present invention is the four Chinese medicine thing altogether, is made as four influence factors; Every herbal medicine is established three magnitudes, i.e. three levels.So 4 factors of employing, 3 horizontal confounding factor uniform designs are got U9 (3 * 3 * 3 * 3) table, obtain 10 prescriptions altogether.Adopt rat MCAT cerebral infarction model, and be contrast with the QINKAILING ZHUSHEYE, to carry out efficacy of medicine observing with 6 effect indexes such as the closely-related iNOS of cerebral ischemia cascade reaction, MDA, TNF-α, IL-1 β, vWF and NSE, the comprehensive relatively pharmacological action intensity of each prescription, and numerical value carries out optimizing by total points system to prescription in view of the above.

The comprehensive advantage analysis of comparing between pharmacodynamics index effect value group shows; the described proportionate relationship of the present composition has all shown good neuroprotective; all the other medication group or DeGrains; though or the improvement effect arranged on some index, on the index the apparent in view effect that increases the weight of to damage is being arranged in addition simultaneously.When the total points queuing is passed judgment on, each pharmacodynamics index value under the same prescription is averaged, each average pharmacodynamics index value summation and the number with same prescription is the total pharmacodynamics effect value of this prescription again; Then, total pharmacodynamics effect value of different prescriptions is sorted according to order from small to large, because various pharmacodynamics index is inhibition type index substantially, so total pharmacodynamics effect value is more little, prescription is excellent more, and the result shows that pharmaceutical composition compatibility group of the present invention is better than the full side of QINGKAILING group.When full side had the attenuation advantage than QINGKAILING, potentiation was also apparent in view at pharmaceutical composition of the present invention in explanation.

Analyze the gained result, determined the proportion compatibility scope that pharmaceutical composition of the present invention is more excellent for comprehensive above-mentioned two kinds.

Experimental example 4 pharmaceutical compositions of the present invention are to the influence of MCAO rat cerebral tissue expression of cell adhesion molecules

1, pharmaceutical composition of the present invention is to the influence of the different period cerebral tissue ICAM-1 expression of cerebral ischemia

As shown in table 1, latter two period of cerebral ischemia, the ICAM-1 of model group rat cerebral tissue expression all is higher than sham operated rats (P＜0.05 or 0.01).Ischemia 12h, pharmaceutical composition group ICAM-1 level of the present invention significantly is lower than model group (P＜0.01), QINGKAILING does not show the effect of preventing the ICAM-1 high expressed significantly, to cerebral ischemia 24h, QINGKAILING and pharmaceutical composition ICAM-1 expression of the present invention and model group compare, the difference nonsignificance.Show that pharmaceutical composition of the present invention can bring into play the effect of ICAM-1 high expressed of preventing extremely in early days at ischemia.

The different period brain tissue homogenate's supernatant ICAM-1 expressions (X ± SD ng/ml) of table 1 ischemia

Group	n	Ischemia 12h	Ischemia 24h
Group	n	Ischemia 12h	Ischemia 24h	Sham operated rats	10	0.688±0.209	1.11±0.369
Model group	10	0.861±0.109 ^△	1.67±0.318 ^△△	Sham operated rats	10	0.688±0.209	1.11±0.369
Model group	10	0.861±0.109 ^△	1.67±0.318 ^△△	Pharmaceutical composition group of the present invention	10	0.598±0.067 ^**	1.53±0.350 ^△△
QINGKAILING	10	0.779±0.113	1.62±0.309 ^△△	Pharmaceutical composition group of the present invention	10	0.598±0.067 ^**	1.53±0.350 ^△△

Annotate: compare with sham operated rats ^△P＜0.05 ^{△ △}P＜0.01

Compare * P＜0.05 * * P＜0.01 with model group

2, pharmaceutical composition of the present invention is to the influence of the different period cerebral tissue E-selectin expression of cerebral ischemia

As shown in table 2, latter two period of cerebral ischemia, the E-selectin of model group rat cerebral tissue expression is higher than sham operated rats (P＜0.01 or 0.05).Ischemia 12h, pharmaceutical composition group cerebral tissue E-selectin level of the present invention is lower than model group (P＜0.01), and the QINGKAILING group does not show prevents expression effect significantly; Ischemia 24h compares with model group, and pharmaceutical composition group of the present invention and QINGKAILING group have all shown prevents expression effect (P＜0.01 or 0.05) significantly, and the former effect is better than the latter.

The different period brain tissue homogenate's supernatant E-selectin expressions (X ± SD ng/ml) of table 2 ischemia

Group	n	Ischemia 12h	Ischemia 24h
Group	n	Ischemia 12h	Ischemia 24h	Sham operated rats model group pharmaceutical composition group of the present invention QINGKAILING group	10 10 10 10	22.86±2.89 26.74±2.59 ^△△ 21.18±2.34 ^** 25.27±3.72 ^△	13.06±2.94 16.87±4.43 ^△ 11.54±1.71 ^** 14.28±2.97 ^*

Annotate: compare with sham operated rats ^△P＜0.05 ^{△ △}P＜0.01

Compare * P＜0.05 * * P＜0.01 with model group

Experimental example 5 pharmaceutical compositions of the present invention are to the influence of MCAO rat aorta regulatory function

1, pharmaceutical composition of the present invention is to the influence of the different period blood plasma ET-1 content of cerebral ischemia.

As shown in table 3, cerebral ischemia 12h and 24h, model group blood plasma ET-1 content significantly increases, and compares P＜0.05 with sham operated rats; Cerebral ischemia 12h, two medication groups are not pointed out the meaningful biological effect, compare P＞0.05 with model group.At cerebral ischemia 24h, pharmaceutical composition group blood plasma ET-1 content of the present invention significantly is lower than model group, P＜0.01, and QINGKAILING does not show useful effect at this moment yet.

The different period blood plasma ET-1 content (X ± SD pg/ml) of table 3 cerebral ischemia

Group	n	Ischemia 12h	Ischemia 24h
Group	n	Ischemia 12h	Ischemia 24h	Sham operated rats model group pharmaceutical composition group of the present invention QINGKAILING group	10 10 10 10	142.26±18.61 159.06±7.66 ^△ 162.69±24.48 ^△ 158.35±6.32 ^△	150.46±22.90 174.30±25.33 ^△ 129.74±21.02 ^**△ 179.20±19.15 ^△

Annotate: compare with sham operated rats ^△P＜0.05 ^{△ △}P＜0.01

Compare * P＜0.05 * * P＜0.01 with model group

2, pharmaceutical composition of the present invention is to the different period blood plasma TXB of cerebral ischemia ₂/ 6-keto-PGF ₁The influence of α ratio

As shown in table 4, cerebral ischemia 12h and 24h, model group blood plasma TXB ₂/ 6-keto-PGF ₁α ratio all is significantly higher than sham operated rats P＜0.01 or 0.05; Ischemia 12h pharmaceutical composition group of the present invention and QINGKAILING group TXB ₂/ 6-keto-PGF ₁α ratio all significantly is lower than model group P＜0.01; Ischemia 24h, pharmaceutical composition group blood plasma TXB of the present invention ₂/ 6-keto-PGF ₁α ratio still significantly is lower than model group P＜0.01, and QINGKAILING group blood plasma TXB ₂/ 6-keto-PGF ₁α ratio then is significantly higher than model group P＜0.05, with model group difference not statistically signigicant.

The different period blood plasma TXB of table 4 cerebral ischemia ₂/ 6-keto-PGF ₁α ratio (X ± SD)

Group	n	Ischemia 12h	Ischemia 24h
Group	n	Ischemia 12h	Ischemia 24h	Sham operated rats model group pharmaceutical composition group of the present invention QINGKAILING group	10 10 10 10	0.350±0.091 0.709±0.168 ^△△ 0.331±0.178 ^ 0.352±0.087 ^	0.366±0.083 0.526±0.204 ^△ 0.251±0.092 ^** 0.704±0.326 ^△

Annotate: compare with sham operated rats ^△P＜0.05 ^{△ △}P＜0.01

Compare * P＜0.05 * * P＜0.01 with model group

Experimental example 6 pharmaceutical compositions of the present invention are to the influence of MCAO rat cerebral tissue inflammatory cytokine content

1, pharmaceutical composition of the present invention is to the influence of ischemic tissue of brain TNF-alpha content

Table 5: the different period TNF-of the brain tissue homogenate α protein contents (X ± SD pg/ml) of cerebral ischemia

Group	n	Ischemia 12h	n	Ischemia 24h
Group	n	Ischemia 12h	n	Ischemia 24h	Sham operated rats model group pharmaceutical composition group of the present invention QINGKAILING group	8 9 10 9	15.241±1.180 17.459±0.261 ^△ 17.747±3.114 ^△ 18.762±0.939 ^△△	10 10 10 10	26.624±1.823 28.324±1.292 ^Δ 24.715±2.843 ^** 25.725±2.841 ^*

Compare with sham operated rats ^△P＜0.05 ^{△ △}P＜0.01

Compare * P＜0.05 * * P＜0.01 with model group

The result shows: cerebral ischemia 12h and 24h hour two time points, model group TNF-α protein content all is significantly higher than sham operated rats p＜0.05, shows that ischemia stimulates the synthetic and secretion that has promoted TNF-α.Each administration group of ischemia 12h and model group more all do not have significant difference.At ischemia 24h, two administration groups all significantly are lower than model group p＜0.01 or 0.05.

2, pharmaceutical composition of the present invention is to the influence of ischemic tissue of brain IL-1 β content

The different period IL-1 of the brain tissue homogenate β protein contents (X ± SD pg/ml) of table 6 cerebral ischemia

Group	n	Ischemia 12h	n	Ischemia 4h
Group	n	Ischemia 12h	n	Ischemia 4h	Sham operated rats model group pharmaceutical composition group of the present invention jasminoidin group	8 9 10 9	0.121±0.021 0.143±0.018 ^△ 0.090±0.048 ^ 0.081±0.040 ^	10 10 10 10	0.191±0.023 0.221±0.042 ^△ 0.150±0.047 ^ 0.149±0.040 ^

Compare with sham operated rats ^△P＜0.05 ^{△ △}P＜0.01,

Compare * P＜0.05 * * P＜0.01 with model group

Table 6 shows, ischemia 12h and 24h, and model group cerebral tissue IL-1 β content is significantly higher than sham operated rats p＜0.05, shows that ischemia has stimulated the synthetic and secretion of IL-1 β.And two medication groups have all shown with model group remarkable significant difference p＜0.01 is arranged relatively by good resistance inhibitor action to this change.

Experimental example 7 pharmaceutical compositions of the present invention are to the protective effect of MCAO rat cerebral tissue blood vessel and neuronal damage

1, pharmaceutical composition of the present invention is to the influence of plasma vWF protein content

The different period plasma vWF content (X ± SD unit) of table 7 cerebral ischemia

Group	n	Ischemia 12h	n	Ischemia 24h
Group	n	Ischemia 12h	n	Ischemia 24h	Sham operated rats model group pharmaceutical composition group of the present invention QINGKAILING group	8 9 10 9	0.028±0.008 0.062±0.017 ^△△ 0.055±0.023 ^△△ 0.068±0.021 ^△△	10 10 10 10	0.024±0.007 0.044±0.013 ^△△ 0.029±0.010 ^** 0.039±0.009 ^△

Compare with sham operated rats ^△P＜0.05 ^{△ △}P＜0.01,

Compare * P＜0.05 * * P＜0.01 with model group

Table 7 shows, ischemia 12h and 24h, and model group plasma vWF content is significantly higher than sham operated rats p＜0.01.Show that ischemia has caused significant injury of vascular endothelial cells.Ischemia 12h, two medication groups and model group are than no remarkable significant difference.Ischemia 24h, pharmaceutical composition group of the present invention significantly is lower than model group p＜0.01, and QINGKAILING group and model group are than no remarkable significant difference.

2, pharmaceutical composition of the present invention is to the influence of NSE contents in serum

The different period NSE contents in serum (X ± SD ng/ml) of table 8 cerebral ischemia

Group	n	Ischemia 12h	n	Ischemia 24h
Group	n	Ischemia 12h	n	Ischemia 24h	Sham operated rats model group pharmaceutical composition group of the present invention QINGKAILING group	6 6 6 8	3.33±0.69 8.72±4.79 ^△ 4.67±1.98 ^* 6.32±2.46 ^△	7 6 9 10	3.44±0.70 5.85±2.34 ^△ 4.58±1.19 ^△ 5.34±2.46 ^△

Compare with sham operated rats ^△P＜0.05 ^{△ △}* P＜0.05 * * P＜0.01 is compared in P＜0.01 with model group

Table 8 shows, cerebral ischemia 12h and 24h, and the model group NSE contents in serum increases, and is remarkable with the sham operated rats comparing difference, P＜0.05.At ischemia 12h, compare with model group, pharmaceutical composition group NSE contents in serum of the present invention significantly reduces P＜0.05.And QINGKAILING group NSE contents in serum is significantly higher than sham operated rats P＜0.05, does not relatively have remarkable reduction with model group.Be extended down to ischemia 24h, compare with model group, two medication group NSE contents in serum all do not have remarkable reduction.

The preparation of embodiment 1 capsule of the present invention

Cholic acid 400g baicalin 450g Fructus Gardeniae 2300g Concha Margaritifera powder 4200g

Be prepared into capsule according to a conventional method.

The preparation of embodiment 2 sprays of the present invention

Cholic acid 740g baicalin 500g Fructus Gardeniae 2800g Concha Margaritifera powder 4500g

Be prepared into spray according to a conventional method.

The preparation of embodiment 3 drop pill of the present invention

Cholic acid 800g baicalin 560g Fructus Gardeniae 3000g Concha Margaritifera powder 5500g

Be prepared into drop pill according to a conventional method.

The preparation of embodiment 4 oral liquids of the present invention

Cholic acid 650g baicalin 550g Fructus Gardeniae 2500g Concha Margaritifera powder 5000g

Be prepared into oral liquid according to a conventional method.

The preparation of embodiment 5 injection tablets of the present invention

Cholic acid 700g baicalin 530g Fructus Gardeniae 2600g Concha Margaritifera powder 4800g

Be prepared into the injection tablet according to a conventional method.

The preparation of embodiment 6 injection of the present invention

Taurine 350g hyodesoxycholic acid 350g baicalin 550g

Fructus Gardeniae 2500g Concha Margaritifera powder 5000g

Make injection according to a conventional method.

The preparation of embodiment 7 lyophilized injectable powders of the present invention

Taurine 480g hyodesoxycholic acid 400g baicalin 450g

Fructus Gardeniae 3300g Concha Margaritifera powder 4300g

Be prepared into lyophilized injectable powder according to a conventional method.

The preparation of embodiment 8 granules of the present invention

Taurine 370g hyodesoxycholic acid 400g baicalin 570g

Fructus Gardeniae 2400g Concha Margaritifera powder 5600g

Be prepared into granule according to a conventional method.

The preparation of embodiment 9 routine tablets of the present invention

Taurine 150g hyodesoxycholic acid 150g baicalin 510g

Fructus Gardeniae 2800g Concha Margaritifera powder 4500g

Be prepared into tablet according to a conventional method.

The preparation of embodiment 10 suspensoids of the present invention

Taurine 350g hyodesoxycholic acid 350g baicalin 500g

Fructus Gardeniae 2700g Concha Margaritifera powder 5000g

Be prepared into suspensoid according to a conventional method.

The preparation of embodiment 11 injection of the present invention

A. get taurine 350g hyodesoxycholic acid 390g baicalin 500g

Fructus Gardeniae 2800g Concha Margaritifera powder 4500g

Make 100 liters of injection as follows.

B. the preparation of gardenia extract, Concha Margaritifera powder hydrolyzed solution

Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, with 70% ethanol percolation of 8 times of amounts, collect percolate, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear paste, add the dilution of 4 times of water gagings, stir evenly, 0-4 ℃ of cold preservation 48 hours, filter, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, it is colourless earlier to be eluted to eluent with distilled water, and 10 times of amounts of reuse, 70% ethanol elution is collected eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with water for injection; Regulate pH value to 6.0～7.0,0-4 ℃ of cold preservation 24 hours, 0.45 μ m filter membrane filters, and filtrate adds the injection water to dissolving, filters with 0.22 μ m filter membrane, gets gardenia extract;

8 times of H that measure 4N of preparation earlier ₂SO ₄Aqueous solution takes by weighing Concha Margaritifera powder again and slowly add (note: constantly stir, the messenger drug hydrorrhea does not go out) under constantly stirring, being heated to boils kept little 6-7 of boiling hour, (constantly adding the deionized water that boils) filters, and filtering residue gets the clear and bright solution of salmon pink with small amount of thermal water washing 2 times; Put cold back adularescent acicular crystals and separate out, refilter, remove crystal, get the salmon pink settled solution; Cooling when clarifying hydrolyzed solution is concentrated an only surplus 2-3 who is equivalent to the raw material total amount and doubly measures adds ethanol and transfers to and contain the alcohol amount and reach 60% cold preservation 24 hours; Sucking filtration, filtrate recycling ethanol takes out cold preservation and uses for dosing to there not being the alcohol flavor.

C. dosing

1, extract the mixing of medicinal liquid:

2, the dissolving of cholic acid:

Measure 75% ethanol (be about cholic acid amount 60 times) and transfer PH to 11, add the deoxycholic acid powder and make whole dissolvings, add the taurine powder and transfer PH to 9 to make it whole dissolvings with 10%NaOH liquid; Mixed extract is mixed in the cholic acid solution, gets admixing medical solutions;

3, the alcohol of admixing medical solutions is handled:

Measuring admixing medical solutions adds 95% ethanol and makes it contain alcohol amount to reach 75%; Transfer PH to 7,2-3 days sucking filtration of sealing cold preservation with 10%NaOH liquid, reclaim ethanol to there not being the alcohol flavor, room temperature is placed; Add fresh water for injection, transfer to neutral 4/5 o'clock, transfer PH to 7.2, cold preservation 3-4 days, paper pulp and sucking filtration, deserved supernatant liquid with 10%NaOH liquid to full dose;

4, the processing of baicalin dissolving and mixing:

Measure the fresh deionized water of total approximately dosing volume, transfer PH to 8, add the baicalin fine powder, transfer PH to 7, it is dissolved fully with 10%NaOH liquid with 10%NaOH liquid; To contain baicalin solution and join in the admixing medical solutions, get total admixing medical solutions.

D. the processing of total admixing medical solutions

Transfer total admixing medical solutions PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, is equipped with embedding with G4 filter bulb sucking filtration and uses; Be prepared into injection by the pharmaceutics conventional method, 10ml/ props up, 0.09 gram/ml, intravenous injection 20-40ml/ day.

The preparation of embodiment 12 injection of the present invention (infusion solutions)

A. get taurine 390g hyodesoxycholic acid 410g baicalin 560g

Fructus Gardeniae 3000g Concha Margaritifera powder 5500g

Make 100 liters of injection infusion solutionses as follows.

Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, with 70% ethanol percolation of 6 times of amounts, collect percolate, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear paste, add the dilution of 5 times of water gagings, stir evenly, 0-4 ℃ of cold preservation 36 hours, filter, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, it is colourless earlier to be eluted to eluent with distilled water, and 10 times of amounts of reuse, 70% ethanol elution is collected eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with water for injection; Regulate pH value to 6.0～7.0,0-4 ℃ of cold preservation 36 hours, 0.45 μ m filter membrane filters, and filtrate adds the injection water to dissolving, filters with 0.22 μ m filter membrane, gets gardenia extract;

8 times of H that measure 4N of preparation earlier ₂SO ₄Aqueous solution; Take by weighing Concha Margaritifera powder again and under constantly stirring, slowly add (attention: constantly stir, the messenger drug hydrorrhea does not go out), being heated to boils kept little 6-7 of boiling hour, (constantly adding the deionized water that boils) filters, and filtering residue gets the clear and bright solution of salmon pink with small amount of thermal water washing 2 times, putting cold back adularescent acicular crystals separates out, refilter, remove crystal, get the salmon pink settled solution; Cooling when clarifying hydrolyzed solution is concentrated an only surplus 2-3 who is equivalent to the raw material total amount and doubly measures adds ethanol and transfers to and contain the alcohol amount and reach 60% cold preservation 24 hours; Sucking filtration, filtrate recycling ethanol takes out cold preservation and uses for dosing to there not being the alcohol flavor.

C. dosing

1, extract the mixing of medicinal liquid:

Gardenia extract, Concha Margaritifera powder hydrolyzed solution are filtered mix homogeneously respectively; Transfer PH to 7 with 10%NaOH liquid, sucking filtration gets mixed extract;

2, the dissolving of cholic acid:

Measure 75% ethanol (be about cholic acid amount 60 times) and transfer PH to 11 with 10%NaOH liquid, add the deoxycholic acid powder and make whole dissolvings, PH～9 added the taurine powder and transferred PH to 9 to make it whole dissolvings this moment; Mixed extract is mixed in the cholic acid solution, gets admixing medical solutions;

3, the alcohol of admixing medical solutions is handled:

4, the processing of baicalin dissolving and mixing:

D. the processing of total admixing medical solutions

Transfer total admixing medical solutions PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, is equipped with embedding with G4 filter bulb sucking filtration and uses; Get total admixing medical solutions, be prepared into infusion solutions according to a conventional method.

The preparation of embodiment 13 capsules of the present invention

A. get taurine 200g hyodesoxycholic acid 200g baicalin 450g

Fructus Gardeniae 2300g Concha Margaritifera powder 4200g

8 times of H that measure 4N of preparation earlier ₂SO ₄Aqueous solution; Take by weighing Concha Margaritifera powder again and slowly add (note: constantly stir, the messenger drug hydrorrhea does not go out) under constantly stirring, being heated to boils kept little 6-7 of boiling hour, and (constantly adding the deionized water that boils) filters, and filtering residue is with small amount of thermal water washing 2 times, the clear and bright solution of salmon pink; Put cold back adularescent acicular crystals and separate out, refilter, remove crystal, get the salmon pink settled solution; Cooling when clarifying hydrolyzed solution is concentrated an only surplus 2-3 who is equivalent to the raw material total amount and doubly measures adds ethanol and transfers to and contain the alcohol amount and reach 60% cold preservation 24 hours; Sucking filtration, filtrate recycling ethanol takes out cold preservation and uses for dosing to there not being the alcohol flavor.

C. dosing

1, extract the mixing of medicinal liquid:

2, the dissolving of cholic acid:

3, the alcohol of admixing medical solutions is handled:

4, the processing of baicalin dissolving and mixing:

D. the processing of total admixing medical solutions

With total admixing medical solutions concentrating under reduced pressure, spraying drying powder-forming, fill is prepared into hard capsule in the hollow capsule.

The preparation of embodiment 14 sprays of the present invention

A. get taurine 350g hyodesoxycholic acid 350g baicalin 530g

Fructus Gardeniae 2600g Concha Margaritifera powder 4800g

Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder, with 70% ethanol percolation of 10 times of amounts, collect percolate, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear paste, add the dilution of 3 times of water gagings, stir evenly, 0-4 ℃ of cold preservation 60 hours, filter, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, it is colourless earlier to be eluted to eluent with distilled water, and 8 times of amounts of reuse, 70% ethanol elution is collected eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with water for injection; Regulate pH value to 6.0～7.0,0-4 ℃ of cold preservation 12 hours, 0.45 μ m filter membrane filters, and filtrate adds the injection water to dissolving, filters with 0.22 μ m filter membrane, gets gardenia extract;

C. dosing

1, extract the mixing of medicinal liquid:

2, the dissolving of cholic acid:

3, the alcohol of admixing medical solutions is handled:

4, the processing of baicalin dissolving and mixing:

D. the processing of total admixing medical solutions

Transfer total admixing medical solutions PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, is equipped with embedding with G4 filter bulb sucking filtration and uses; Get total admixing medical solutions, be prepared into injection; Injection is evaporated to 1/2 amount, and every 10ml fill is in the mini spray bottle; Be prepared into 5000 of sprays, consumption per day is 3-5ml.

The preparation of embodiment 15 drop pill of the present invention

A. get taurine 300g hyodesoxycholic acid 350g baicalin 550g

Fructus Gardeniae 2500g Concha Margaritifera powder 5000g

Get the Fructus Gardeniae of recipe quantity, be ground into coarse powder,, with 70% ethanol percolation of 8 times of amounts, collect percolate, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear paste, add the dilution of 4 times of water gagings, stir evenly, 0-4 ℃ of cold preservation 48 hours, filter, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, it is colourless earlier to be eluted to eluent with distilled water, and 10 times of amounts of reuse, 70% ethanol elution is collected eluent, decompression recycling ethanol also concentrates, and drying gets the Fructus Gardeniae semi-finished product; Get the Fructus Gardeniae semi-finished product and dissolve, filter with water for injection; Regulate pH value to 6.0～7.0,0-4 ℃ of cold preservation 24 hours, 0.45 μ m filter membrane filters, and filtrate adds the injection water to dissolving, filters with 0.22 μ m filter membrane, gets gardenia extract;

C. dosing

1, extract the mixing of medicinal liquid:

2, the dissolving of cholic acid:

3, the alcohol of admixing medical solutions is handled:

4, the processing of baicalin dissolving and mixing:

D. the processing of total admixing medical solutions

Transfer total admixing medical solutions PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, is equipped with embedding with G4 filter bulb sucking filtration and uses; With total admixing medical solutions concentrating under reduced pressure, spraying drying powder-forming; (ratio is PEG4000: stearic acid: paraffin: medicated powder=3.60: 0.32: 0.08: 1.00) water-bath is dissolved with this medicated powder and stroma ground substance, mix homogeneously, use internal diameter 1.69mm, the dropper of external diameter 3.81mm under 75 ℃ with the speed of dripping about 48/min, splash into from top to bottom in the liquid paraffin liquid coolant, collect drop pill and absorb coolant promptly with filter paper; Be prepared into drop pill.

The preparation of embodiment 16 freeze-dried powders of the present invention

A. get taurine 300g hyodesoxycholic acid 430g baicalin 510g

Fructus Gardeniae 2800g Concha Margaritifera powder 4500g

Make freeze-dried powder as follows.

C. dosing

1, extract the mixing of medicinal liquid:

2, the dissolving of cholic acid:

3, the alcohol of admixing medical solutions is handled:

4, the processing of baicalin dissolving and mixing:

D. the processing of total admixing medical solutions

Transfer total admixing medical solutions PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, makes freeze-dried powder through lyophilization.

The preparation of embodiment 17 oral liquids of the present invention

A. get taurine 200g hyodesoxycholic acid 400g baicalin 500g

Fructus Gardeniae 2700g Concha Margaritifera powder 5000g

C. dosing

1, extract the mixing of medicinal liquid:

2, the dissolving of cholic acid:

3, the alcohol of admixing medical solutions is handled:

4, the processing of baicalin dissolving and mixing:

D. the processing of total admixing medical solutions

Transfer total admixing medical solutions PH to 7.2, add activated carbon (0.3) water-bath and boil, filtered while hot, filtrate sealing cold preservation 3-4 days, paper pulp also filters, and transfers PH to 7.2, is equipped with embedding with G4 filter bulb sucking filtration and uses; Total admixing medical solutions is prepared into oral liquid according to a conventional method.

The preparation of embodiment 18 injection tablets of the present invention

A. get taurine 320g hyodesoxycholic acid 480g baicalin 470g

Fructus Gardeniae 3000g Concha Margaritifera powder 4000g

C. dosing

1, extract the mixing of medicinal liquid:

2, the dissolving of cholic acid:

3, the alcohol of admixing medical solutions is handled:

4, the processing of baicalin dissolving and mixing:

D. the processing of total admixing medical solutions

To always mix concentratedly, be prepared into the injection tablet according to a conventional method.

Claims

1. A pharmaceutical composition, characterized in that it is made from the following raw materials by weight:

Cholic acid 3-9 parts by weight Baicalin 4-6 parts by weight

20-35 parts by weight of gardenia and 40-60 parts by weight of mother-of-pearl powder.

2. The pharmaceutical composition according to claim 1, characterized in that it is made of the following raw materials in parts by weight:

Cholic acid 6.5-7.4 parts by weight Baicalin 5.0-5.5 parts by weight

25.0-28.0 parts by weight of gardenia and 45.0-50.0 parts by weight of mother-of-pearl powder.

3. The pharmaceutical composition according to claim 1, characterized in that it is made of the following raw materials in parts by weight:

Cholic acid 4 parts by weight Baicalin 4.5 parts by weight

23 parts by weight of gardenia and 42 parts by weight of mother-of-pearl powder.

4. The pharmaceutical composition according to claim 1, characterized in that it is made of the following raw materials in parts by weight:

Cholic acid 8 parts by weight Baicalin 5.6 parts by weight

30 parts by weight of gardenia, 55 parts by weight of mother-of-pearl powder.

5. The pharmaceutical composition according to claim 1, characterized in that it is made of the following raw materials in parts by weight:

Cholic acid 7.4 parts by weight Baicalin 5.0 parts by weight

28 parts by weight of gardenia, 45 parts by weight of mother-of-pearl powder.

6. The pharmaceutical composition according to claim 1, characterized in that it is made of the following raw materials in parts by weight:

Cholic acid 6.5 parts by weight Baicalin 5.5 parts by weight

25 parts by weight of gardenia and 50.0 parts by weight of mother-of-pearl powder.

7. The pharmaceutical composition according to claim 1, characterized in that it is made of the following raw materials in parts by weight:

Cholic acid 7 parts by weight Baicalin 5.3 parts by weight

26 parts by weight of gardenia and 48 parts by weight of mother-of-pearl powder.

8. The pharmaceutical composition according to any one of claims 1-7, characterized in that said cholic acid is composed of taurocholic acid and isodeoxycholic acid in a ratio of 2-4:2-4.5.

9. The pharmaceutical composition according to claim 8, characterized in that said cholic acid is composed of taurocholic acid and isodeoxycholic acid in a ratio of 3.0-3.5:3.5-3.9.

10. The pharmaceutical composition as claimed in claim 8, characterized in that said cholic acid is composed of taurocholic acid and isodeoxycholic acid in a ratio of 2.2:3.4.

11. The pharmaceutical composition as claimed in claim 8, characterized in that said cholic acid is composed of taurocholic acid and isodeoxycholic acid in a ratio of 3.9:4.1.

12. The pharmaceutical composition according to claim 8, characterized in that said cholic acid is composed of taurocholic acid and isodeoxycholic acid in a ratio of 3.2:3.8.

13. A pharmaceutical composition according to claim 1, 2, 3, 4, 5, 6, 7, 9, 10, 11 or 12, characterized in that it is prepared into a clinically acceptable dosage form according to conventional methods: injection preparation , tablets, capsules, sprays, pills, granules, suspensions, dropping pills or oral liquid preparations.

14. The pharmaceutical composition according to claim 8, characterized in that it is prepared into clinically acceptable dosage forms according to conventional methods: injection preparations, tablets, capsules, sprays, pills, granules, suspensions, dropping pills or Oral liquid formulation.

15. The preparation method of the pharmaceutical composition as claimed in claim 8, characterized in that the method comprises the following steps:

A. Take taurocholic acid, isodeoxycholic acid, baicalin, gardenia, mother-of-pearl powder;

B. Prepare gardenia extract and mother-of-pearl powder hydrolyzate, mix uniformly, adjust pH to 7, obtain mixed extract;

C. Take ethanol, adjust the pH to 11, add deoxycholic acid powder to make it all dissolve, add tauric acid powder to adjust the pH to 9 to make it completely dissolved; mix the mixed extract solution in the cholic acid solution to obtain a mixed medicinal solution; Precipitate the mixed medicinal solution with ethanol, refrigerate, suction filter, and recover ethanol; add water for injection, add baicalin fine powder, and make it completely dissolve to obtain the total mixed medicinal solution;

D. Take the total mixed medicinal solution, add auxiliary materials, and make injection preparations, tablets, capsules, sprays, pills, granules, suspensions, dropping pills or oral liquid preparations according to conventional preparation processes or steps.

16. The preparation method of the pharmaceutical composition as claimed in claim 15, wherein the preparation method of the gardenia extract is:

Take gardenia, crush it into coarse powder, percolate with 60% to 80% ethanol in an amount of 6 to 10 times, collect the percolation liquid, recover ethanol, concentrate to a clear paste with a relative density of 1.10 at 60°C, add 3 to 5 Dilute with twice the amount of water, stir well, refrigerate at 0-4°C for 36-60 hours, filter, concentrate the filtrate under reduced pressure to a clear paste with a relative density of 1.10 at 60°C, put it on an activated carbon column, and first elute with distilled water to the eluent Colorless, then elute with 8 to 12 times the amount of 60% to 80% ethanol, collect the eluent, recover the ethanol under reduced pressure, concentrate, and dry to obtain the semi-finished product of Gardenia; dissolve the semi-finished product of Gardenia with water for injection, and filter; Adjust the pH value to 6.0-7.0, refrigerate at 0-4°C for 12-36 hours, filter through a 0.45 μm filter, add water for injection to the filtrate until dissolved, and filter through a 0.22 μm filter to obtain a gardenia extract.

17. The preparation method of the pharmaceutical composition as claimed in claim 16, wherein the preparation method of the gardenia extract is:

Take gardenia, crush it into coarse powder, percolate with 8 times the amount of 70% ethanol, collect the percolation liquid, recover ethanol, concentrate to a clear paste with a relative density of 1.10 at 60°C, add 4 times the amount of water to dilute, and stir well , refrigerated at 0-4°C for 48 hours, filtered, the filtrate was concentrated under reduced pressure to a clear paste with a relative density of 1.10 at 60°C, put it on an activated carbon column, and eluted with distilled water until the eluent was colorless, and then used 10 times the amount of 70 % ethanol for elution, collect the eluent, recover the ethanol under reduced pressure and concentrate, dry to obtain the semi-finished product of Gardenia; dissolve the semi-finished product of Gardenia with water for injection, filter; adjust the pH value to 6.0-7.0, and refrigerate at 0-4°C for 24 hours, filter through a 0.45 μm filter, add water for injection to the filtrate until dissolved, and filter through a 0.22 μm filter to obtain a gardenia extract.

18. The preparation method of the pharmaceutical composition as claimed in claim 15, 16 or 17, characterized in that the preparation method of the mother-of-pearl powder hydrolyzate is:

First prepare 8 times the amount of 4N H ₂ SO ₄ aqueous solution; add mother-of-pearl powder, heat to boil and keep boiling slightly for 6-7 hours, filter, and wash the filter residue with hot water twice to obtain an orange-red clear solution; after cooling, white needles Crystals are precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; the clarified hydrolyzed solution is concentrated to only 2-3 times the amount of the total amount of raw materials, cooled, and ethanol is added to adjust the alcohol content to 60%. Refrigerate for 24 hours; filter with suction, recover ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for liquid preparation.

19. The preparation method of any one of the pharmaceutical compositions according to claims 9-12, characterized in that the method comprises the following steps:

20. The preparation method of the pharmaceutical composition as claimed in claim 19, wherein the preparation method of the gardenia extract is:

21. The preparation method of the pharmaceutical composition as claimed in claim 20, wherein the preparation method of the gardenia extract is:

22. The preparation method of the pharmaceutical composition as claimed in claim 19, wherein the preparation method of the mother-of-pearl powder hydrolyzate is:

23. The preparation method of the pharmaceutical composition as claimed in claim 8, characterized in that the method comprises the following steps:

B. Get Fructus Gardeniae, be ground into coarse powder, percolate with 70% ethanol of 8 times of amount, collect percolation liquid, reclaim ethanol, be concentrated to the clear cream that relative density is 1.10 under 60 ℃, add 4 times of amount of water to dilute, Stir well, refrigerate at 0-4°C for 48 hours, filter, and concentrate the filtrate under reduced pressure to a clear paste with a relative density of 1.10 at 60°C, put it on an activated carbon column, and elute with distilled water until the eluent is colorless, then use 10 times Elute with 70% ethanol, collect the eluent, recover the ethanol under reduced pressure and concentrate, dry to obtain the semi-finished product of Gardenia; dissolve the semi-finished product of Gardenia with water for injection, filter; adjust the pH value to 6.0-7.0, 0-4 °C Refrigerate for 24 hours, filter through a 0.45 μm filter membrane, add water for injection to the filtrate until dissolved, and filter through a 0.22 μm filter membrane to obtain the gardenia extract;

First prepare 8 times the amount of 4N H ₂ SO ₄ aqueous solution; add mother-of-pearl powder, heat to boil and keep boiling slightly for 6-7 hours, filter, and wash the filter residue with hot water twice to obtain an orange-red clear solution; after cooling, white needles Crystals are precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; the clarified hydrolyzed solution is concentrated to only 2-3 times the amount of the total amount of raw materials, cooled, and ethanol is added to adjust the alcohol content to 60%. Refrigerate for 24 hours; filter with suction, recover ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for liquid preparation;

C. Filter the gardenia extract and the mother-of-pearl powder hydrolyzate respectively, mix well, adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract; get 75% ethanol that is 60 times the amount of cholic acid, and use Adjust the pH to 11 with 10% NaOH solution, add deoxycholic acid powder to dissolve it all, add taurocholic acid powder to adjust the pH to 9 to dissolve it completely; mix the mixed extract with the cholic acid solution to obtain a mixed medicinal solution; mix Add 95% ethanol to the liquid medicine to make the alcohol content reach 75%; use 10% NaOH solution to adjust the pH to 7, seal and refrigerate for 2-3 days and filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add fresh water for injection , adjusted to neutral to 4/5 of the full amount, adjust the pH to 7.2 with 10% NaOH solution, refrigerate for 3-4 days, and filter with suction to obtain a clear liquid; take the fresh deionized water of the total liquid volume, and use 10% Adjust the pH to 8 with NaOH solution, add baicalin fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin to the mixed medicinal solution to obtain the total mixed medicinal solution;

D. Adjust the pH of the mixed medicinal solution to 7.2, add activated carbon to boil in a water bath, filter while it is hot, seal and refrigerate the filtrate for 3-4 days, and filter the pulp, adjust the pH to 7.2, and use G4 filter ball to prepare for filling and sealing. into injection preparations.

24. The preparation method of any one of the pharmaceutical compositions according to claims 9-12, characterized in that the method comprises the following steps:

B. Get Fructus Gardeniae, be ground into coarse powder, percolate with 70% ethanol of 6 times of amount, collect percolation liquid, reclaim ethanol, be concentrated to the clear ointment that relative density is 1.10 under 60 ℃, add 5 times of amount of water to dilute, Stir well, refrigerate at 0-4°C for 36 hours, filter, and concentrate the filtrate under reduced pressure to a clear paste with a relative density of 1.10 at 60°C, put it on an activated carbon column, and elute with distilled water until the eluent is colorless, then use 10 times Elute with 70% ethanol, collect the eluent, recover the ethanol under reduced pressure and concentrate, dry to obtain the semi-finished product of Gardenia; dissolve the semi-finished product of Gardenia with water for injection, filter; adjust the pH value to 6.0-7.0, 0-4 °C Refrigerate for 36 hours, filter with a 0.45 μm filter, add water for injection to the filtrate until dissolved, and filter with a 0.22 μm filter to obtain the gardenia extract;

25. The use of the pharmaceutical composition according to claim 1, 2, 3, 4, 5, 6, 7, 9, 10, 11 or 12 in the preparation of a medicament for treating cerebral ischemia.

26. The use of the pharmaceutical composition as claimed in claim 8 in the preparation of medicaments for treating cerebral ischemia.

27. The application according to claim 25, wherein the treatment of cerebral ischemia refers to suppressing the high expression of ICAM-1, suppressing the expression of E-selectin in brain tissue, reducing plasma ET-1 content, reducing plasma TXB ₂ /6-keto - PGF ₁ α ratio, decrease the synthesis and secretion of TNF-α, inhibit the synthesis and secretion of IL-1β, reduce the plasma vWF content or reduce the serum NSE content.

28. The application according to claim 26, wherein the treatment of cerebral ischemia refers to suppressing the high expression of ICAM-1, suppressing the expression of E-selectin in brain tissue, reducing plasma ET-1 content, reducing plasma TXB ₂ /6-keto - PGF ₁ α ratio, decrease the synthesis and secretion of TNF-α, inhibit the synthesis and secretion of IL-1β, reduce the plasma vWF content or reduce the serum NSE content.