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

Abstract

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 pharmaceutical composition for treating cerebral apoplexy and its preparation method

technical field

The invention relates to a pharmaceutical composition and a preparation method thereof, in particular to a pharmaceutical composition for treating cerebral apoplexy and a preparation method thereof, belonging to the technical field of medicine.

Background technique

Qingkailing Injection is a traditional Chinese medicine compound injection that is scientifically developed and purified from the classic Chinese medicine prescription Angong Niuhuang Pills. It has the effects of clearing heat and detoxifying, resolving phlegm and dredging collaterals, refreshing the mind and resuscitating the mind. Clinically, it is mainly used for high fever, coma, stroke hemiplegia, acute and chronic hepatitis, upper respiratory tract infection, pneumonia and other diseases. Over the past 22 years of clinical application, due to its obvious curative effect, the application has been increasing day by day. Since 1992, it has been approved as the first-choice essential drug for the emergency department of Chinese medicine hospitals in the country for three consecutive times, and it is also listed as "National Traditional Chinese Medicine Protected Variety". Especially in the past 10 years, its unique curative effect on cerebrovascular diseases has been widely recognized clinically.

However, in recent years, adverse reactions of Qingkailing injection have occurred frequently, and related reports have increased day by day, and some serious adverse reactions have even occurred, leading to the death of patients. The main adverse reactions include: allergic reactions (rash, itching, exfoliative dermatitis, angioneurotic edema, anaphylactic shock, etc.), central nervous system adverse reactions (disorder of consciousness, coma, convulsions, mental confusion, etc.) and occasional nausea and vomiting , abdominal pain, phlebitis, acute renal failure, etc. Therefore, it is imperative to improve the production process, increase the purity of the drug, minimize the content of impurities without pharmacological effects, and reduce adverse reactions.

Since Qingkailing Injection is a compound injection of traditional Chinese medicine, it has a wide range of efficacy and treats many diseases, and the material basis of the efficacy of each medicine is not clear, so it is difficult to improve the process. Therefore, it is a feasible way to study the formula and process of Qingkailing injection for the main diseases.

Contents of the invention

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

The purpose of the present invention is achieved through the following technical solutions:

The pharmaceutical composition of the present invention is:

3-9 parts by weight of cholic acid, 4-6 parts by weight of baicalin, 20-35 parts by weight of gardenia, 40-60 parts by weight of mother-of-pearl powder; the preferred range of the above pharmaceutical composition is: 6.5-7.4 parts by weight of cholic acid, 5.0-5.5 parts by weight of baicalin, 25.0-28.0 parts by weight of gardenia, 45.0-50.0 parts by weight of mother-of-pearl powder;

The preferred ratio of the above pharmaceutical composition is: 4 parts by weight of cholic acid, 4.5 parts by weight of baicalin, 23 parts by weight of gardenia, 42 parts by weight of mother-of-pearl powder; 8 parts by weight of cholic acid, 5.6 parts by weight of baicalin, 30 parts by weight, 55 parts by weight of mother-of-pearl powder; 7.4 parts by weight of cholic acid, 5.0 parts by weight of baicalin, 28 parts by weight of gardenia, 45 parts by weight of mother-of-pearl powder; 6.5 parts by weight of cholic acid, 5.5 parts by weight of baicalin, gardenia 25.0 parts by weight, 50.0 parts by weight of mother-of-pearl powder; or 7 parts by weight of cholic acid, 5.3 parts by weight of baicalin, 26 parts by weight of gardenia, and 48 parts by weight of mother-of-pearl powder.

In the above pharmaceutical composition, the cholic acid is composed of taurocholic acid and isodeoxycholic acid in a ratio of 2-4:2-4.5; wherein the preferred ratio range of taurocholic acid and isodeoxycholic acid is: 3.0-3.5:3.5-3.9; the preferred ratio of taurocholic acid and isodeoxycholic acid is: 2.2:3.4, 3.9:4.1 or 3.2:3.8.

In the above pharmaceutical composition, the baicalin can be replaced by baicalin, norbaicalin, wogonin or wogonin.

The above-mentioned pharmaceutical composition is prepared into various clinically acceptable dosage forms according to conventional pharmaceutical methods, including but not limited to one of the following dosage forms: injection preparations, tablets, capsules, sprays, pills, granules, Suspensions, dripping pills, oral liquid preparations, etc.; the injection preparations can be injections, large infusions, freeze-dried powder injections, tablets for injection, suspension injections, etc.

The preparation process of the pharmaceutical composition of the present invention comprises the following steps:

A. Take taurocholic acid, isodeoxycholic acid, baicalin, gardenia, mother-of-pearl powder according to the prescription;

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 clinically acceptable dosage forms according to conventional preparation processes or steps: injection preparations, tablets, capsules, sprays, pills, granules, suspensions, drop pills, Oral liquid preparations, etc.

As mentioned above, the preparation method of the pharmaceutical composition of the present invention, wherein the gardenia extract can be prepared by the following method:

Get the Fructus Gardeniae of prescription quantity, be ground into coarse powder, according to the percolation method ("Chinese Pharmacopoeia" 2000 edition one appendix IO) under the liquid extract item, percolation with 60%～80% ethanol of 6～10 times amount , 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 times the amount of water to dilute, stir well, refrigerate at 0-4°C for 36 to 60 hours, filter, and decompress the filtrate Concentrate 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 until the eluent is colorless, then elute with 8 to 12 times the amount of 60% to 80% ethanol, collect the eluent, Recover 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, and 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 Gardenia extract.

As mentioned above, the preparation method of the pharmaceutical composition of the present invention, wherein the mother-of-pearl powder hydrolyzate can be prepared by the following method:

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;

As mentioned above, the preparation method of the pharmaceutical composition of the present invention, wherein said liquid preparation step can be specifically as follows:

1. Mixing of extraction liquid

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, mix them evenly, adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid

Take 75% ethanol that is 60 times the amount of cholic acid, adjust the pH to 11 with 10% NaOH solution, add deoxycholic acid powder to dissolve it completely, add tauric acid powder to adjust the pH to 9 to dissolve it completely; mix and extract The solution is mixed in the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine

Add 95% ethanol to the mixed medicinal solution to make the alcohol content reach 75%; adjust the pH to 7 with 10% NaOH solution, 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 injection Adjust the pH to 7.2 with 10% NaOH solution when it is neutralized to 4/5 of the full amount with water, refrigerate for 3-4 days, and filter with suction to obtain a clear liquid;

4. Dissolution and mixing of baicalin

Get about the fresh deionized water of total dosing volume, adjust PH to 8 with 10% NaOH solution, add baicalin fine powder, adjust PH to 7 with 10% NaOH solution, make it dissolve completely; Add baicalin-containing solution to In the mixed medicinal liquid, the total mixed medicinal liquid is obtained.

The preparation process of the pharmaceutical composition injection preparation of the present invention comprises the following steps:

Prepare the total mixed medicinal solution according to the above preparation method, adjust the pH to 7.2, add activated carbon (0.3) to boil in a water bath, filter while hot, seal the filtrate and refrigerate for 3-4 days, pulp and filter, adjust the pH to 7.2, and use G4 filter ball Suction filtration to make injection preparations.

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

Prepare the total mixed medicinal solution according to the above preparation method, adjust the pH to 7.2, add activated carbon (0.3) to boil in a water bath, filter while hot, seal the filtrate and refrigerate for 3-4 days, pulp and filter, adjust the pH to 7.2, and use G4 filter ball Suction filtration, concentration under reduced pressure, spray-drying into powder; this medicinal powder and matrix material (ratio is PEG4000: stearic acid: paraffin: medicinal powder=3.60: 0.32: 0.08: 1.00) water bath melts, mixes evenly, with dropper at 75 ℃, drop into the liquid paraffin cooling liquid from top to bottom, collect the dropping pills and absorb the cooling agent with filter paper, and obtain the dropping pills of the pharmaceutical composition of the present invention.

The effective components of the composition of the present invention include cholic acids (taucholic acid, hyocholic acid and isodeoxycholic acid, etc.), baicalins (baicalin is the main ingredient, also wogonin, scutellarin, etc.), Gardenia iridoids (mainly geniposide, dehydroxygeniposide and gardenic acid, etc.), pigments (crocin, crocetin, etc.), bound calcium and free calcium, etc.

The injection prepared by the preparation method described in the technical solution of the present invention has the lightest color compared with Qingkailing injection and the injection prepared by changing the hydrolysis method of mother of pearl, and its total solid content is less . Adopt the Waster rat MCAT cerebral infarction model, and take Qingkailing injection as contrast, measure brain tissue SOD, MDA, NOS content etc.; The result shows, for the treatment of cerebral ischemic injury, composition of the present invention is on some indexes with Qingkailing is basically similar, and the effect of the injection prepared by the method described in the technical scheme is particularly obvious, and it is better than Qingkailing in some aspects in some proportions.

According to relevant experiments, the results show that: the new pharmaceutical composition of the present invention and its preparation process not only eliminates the medicinal taste that may cause adverse reactions, but also reduces adverse reactions, and minimizes the content of impurities without pharmacological effects; Four kinds (of which taurocholic acid and isodeoxycholic acid are collectively referred to as cholic acid) have improved the precision of the gardenia extract; and its biological effect has no effect on the protective effect of the original Qingkailing injection on cerebral ischemic injury. The statistically significant difference basically maintains the brain protection effect of Qingkailing injection in the pathological process of cerebral ischemic injury, and at the same time removes the components that may cause adverse reactions, achieving the optimization goal of stable effect and attenuation. Compared with Qingkailing, the pharmaceutical composition of the present invention has the following advantages in terms of pharmacodynamics: ① The pharmaceutical composition of the present invention can inhibit the microvascular cell adhesion process initiated by cerebral ischemia, thereby improving the microcirculation perfusion of ischemic tissue and improving ischemia It has a more significant advantage in the hypoxic state; ② the pharmaceutical composition of the present invention is more significant in reducing the damage of ischemia to neurons and vascular endothelial cells.

The following experimental examples are used to further illustrate but not limit the present invention.

Experimental example 1 Study on the characteristics of Qingkailing medicinal substances and compatibility in animal models of cerebral ischemia injury

Focal cerebral ischemia-reperfusion model of middle cerebral artery infarction in rats using suture method, and incomplete cerebral ischemia injury animal model of bilateral common carotid arteries in rats with repeated clipping, and overall evaluation of neurological deficit and mortality , combined with the biochemical analysis of a series of metabolic damage indicators, the screening analysis of the effective components in the active substances of Qingkailing injection was carried out.

A. Overall effect: including cholic acid, baicalin, gardenia extract, honeysuckle extract, Banlangen extract, mother of pearl and buffalo horn hydrolyzate and other medicinal substances of Qingkailing, all of which can significantly improve the neurological function of model animals Symptoms of dysfunction. There is a significant synergistic effect between each part, and the effect of Qingkailing Quanfang is the most remarkable.

Cholic acid and baicalin can reduce the mortality of model animals, while the other single components have no significant effect on the mortality of model animals or have a tendency to aggravate. The mortality rate of each compatibility group decreased significantly, and with the increase of the compatibility of various effective substances, the mortality rate further decreased, and the mortality rate of Qingkailing Quanfang was zero, showing a good compatibility effect of reducing toxicity and synergizing.

B. Anti-lipid peroxidation: Honeysuckle, Banlangen, mother of pearl and buffalo horn can significantly increase the SOD level of the brain tissue of model animals, while cholic acid, baicalin, and gardenia have no effect on improving the SOD level of the brain tissue. The compatibility of baicalin and gardenia produced a significant effect of increasing the level of SOD in the brain tissue of model animals, while the other combinations did not show synergistic effect. Banlangen can significantly reduce the LPO content in the brain tissue of model animals, while other drugs have no significant effect on this. However, the combination of baicalin and gardenia can significantly reduce the LPO content of brain tissue. Compatibility can significantly reduce the LPO content of brain tissue. However, baicalin and honeysuckle had no significant effect on the process of lipid peroxidation.

C. Effects on brain energy metabolism: gardenia, banlangen, mother of pearl and buffalo horn can significantly reduce the activity of sodium-potassium-ATPase in the brain tissue of model animals, baicalin has the opposite effect, while cholic acid and honeysuckle have no significant effect . Compatibility of cholic acid with mother of pearl and buffalo horn, as well as compatibility of baicalin and gardenia, can weaken the latter's effect on reducing the activity of sodium-potassium-ATPase in brain tissue, suggesting that the two groups of compatibility have an effect on sodium-potassium-ATPase. There may be an antagonistic effect on the activity. The compatibility of honeysuckle and banlangen did not show obvious synergistic effect.

Mother of pearl and buffalo horn have a significant improvement trend on brain energy charge, cholic acid, baicalin, and gardenia have no significant effect, and honeysuckle and banlangen have a certain improvement trend. Mother of pearl, buffalo horn and cholic acid still maintain the original effect, while baicalin and gardenia, honeysuckle and banlangen have no significant effect on the change of brain energy charge, mother of pearl, buffalo horn and baicalin, gardenia , honeysuckle and Banlangen had a negative impact on their original effects.

D. Analysis of damage response indicators: Banlangen can significantly reduce the level of vWF in the plasma of model animals, while cholic acid, baicalin, gardenia, honeysuckle, mother of pearl and buffalo horn have no significant effect on improving the level of vWF in model animals, and cholic acid even will have a negative impact. However, the combination of cholic acid with mother of pearl and buffalo horn, and the combination of baicalin and gardenia all produced a significant effect of reducing the plasma vWF level of model animals, and the two combinations had a synergistic effect, while the combination of honeysuckle and banlangen did not appear. Significant synergistic effect, the effect of Qingkailing Quanfang is more significant. Bile acid and baicalin can significantly reduce the NOS level in the brain tissue of model animals, while gardenia, honeysuckle, banlangen, mother of pearl and buffalo horn have no effect on improving the NOS level in brain tissue of model animals. Compatibility did not show obvious synergistic effect.

The above research results show that the components of Qingkailing injection have relatively specific effects on different pathological links of cerebral ischemic injury, and there are many effect relationships such as effective and ineffective, compatibility synergy and attenuation.

Experimental Example 2 Application of Cell Model to Screening of Qingkailing Pharmacological Substances

Using the cultured neurons, glial cells and endothelial cells, the drug effect substance screening of the cell model was carried out under the non-toxic dosage to the normal cultured cells.

A. Effects on neurons: Qingkailing Quanfang has obvious protective effect on neurons at a certain dose. Among them, the mixture of gardenia, isatis root, mother of pearl and buffalo horn hydrolyzate has obvious protective effect on neurons, promotes the proliferation of neurons (SH-SY5Y), inhibits the apoptosis of neurons, and inhibits lipid peroxides increase the activity of SOD and NOS in the anti-lipid peroxidation system, and increase the concentration of NO; the mixed solution of taurocholic acid, hyocholic acid, gardenia, isatidis, mother of pearl and buffalo horn hydrolyzate can produce neurons It has obvious toxic effect, which inhibits the apoptosis of neurons, inhibits the production of lipid peroxide, increases the activity of free radical scavengers SOD and NOS, and increases the concentration of NO. The mixed solution of buffalo horn hydrolyzate decreased to some extent. The study of a single drug shows that: at a certain dose, taurocholic acid and gardenia have obvious protective effects on neurons, while hyocholic acid and baicalin have a high toxic effect on neurons, but high doses of baicalin have no effect on SH. The inhibitory effect of -SY5Y neurons is mild in the lower dose group, and the low dose of honeysuckle, mother of pearl and buffalo horn hydrolyzate and large dose of Radix isatidis have obvious protective effects on neurons. Therefore, the compatibility of different doses and the compatibility of Qingkailing medicinal substances in different groups can reduce the deficiency of each single herbal medicine and achieve the best efficacy of Qingkailing.

B. Effect on endothelial cells: Qingkailing Quanfang has obvious protective effect on endothelial cells at a certain dose. Among them, the mixed solution of gardenia, isatidis, mother of pearl and buffalo horn hydrolyzate has obvious protective effect on endothelial cells and promotes the proliferation of endothelial cells (ECV304); taurocholic acid, hyocholic acid, gardenia, isatidis, mother of pearl and The mixed solution of buffalo horn hydrolyzate can promote the proliferation of endothelial cells, which is weaker than that of gardenia, isatis root, mother of pearl and buffalo horn hydrolyzate, but the toxicity is increased. The research of single components shows that taurocholic acid, baicalin, mother of pearl and buffalo horn hydrolyzate can significantly promote the proliferation of endothelial cells, and large doses of gardenia and hyocholic acid also have a strong effect on promoting the proliferation of endothelial cells. Honeysuckle and Banlangen have obvious inhibitory effect on the growth of endothelial cells.

C. Effects on glial cells: Qingkailing formula has obvious inhibitory effect on glial cells, and the inhibitory effect of low-dose Qingkailing whole solution on glial cells is significantly lower than that of high-dose. Among them, the mixture of gardenia, isatis root, mother of pearl and buffalo horn hydrolyzate has a significant protective effect on glial cells. The mixture of taurocholic acid, hyocholic acid, gardenia, isatis root, mother of pearl and buffalo horn hydrolyzate has a significant inhibitory effect on glial cells. Low doses of taurocholic acid, hyocholic acid, and baicalin have significant protective effects on glial cells, and high doses of taurocholic acid, hyocholic acid, and baicalin have a higher inhibitory effect on glial cells. Gardenia, honeysuckle, isatis root, mother of pearl and buffalo horn hydrolyzate have mild inhibitory effect on glial cells.

In view of the characteristics of clinical adverse reactions of Qingkailing injection, first of all, based on the principle of maintaining efficacy and reducing toxicity, it is determined that the adverse reactions and insignificant effects of Qingkailing are buffalo horn, honeysuckle and isatis root, which are effective prescriptions for the treatment of cerebral ischemic injury It is the pharmaceutical composition of the present invention.

Experimental Example 3 Optimal Research on the Compatibility Changes of the Pharmaceutical Composition of the Present Invention

The pharmaceutical composition of the present invention has four medicines in total, which are set as four influencing factors; each medicine has three orders of magnitude, namely three levels. Therefore, the 4-factor 3-level mixed factor uniform design method was adopted, and the U9 (3×3×3×3) table was used to obtain a total of 10 prescriptions. Rat MCAT cerebral infarction model was used, and Qingkailing injection was used as the control, and six effect indicators, which were closely related to the cerebral ischemic cascade reaction, including iNOS, MDA, TNF-α, IL-1β, vWF and NSE, were tested. Observe the drug effect, comprehensively compare the pharmacological action strength of each formula, and optimize the formula through the total score system based on this value.

The comprehensive superiority analysis of the group comparison of the efficacy index effect value shows that the proportional relationship described by the composition of the present invention has shown a good neuroprotective effect, and the remaining drug groups may have no obvious effect, or although some indicators have improved effect, but at the same time, it has a more obvious effect of aggravating damage on other indicators. When the total score is judged in line, the drug efficacy index values under the same group of prescriptions are averaged, and then the average drug efficacy index values of the same group of prescriptions are summed, and the sum is the total pharmacological effect value of the group prescription; The total pharmacological effect value of the prescription is sorted according to the order from small to large, because various drug effect indexes are basically inhibitory indexes, so the smaller the total pharmacological effect value, the better the prescription. The results show that the pharmaceutical composition of the present invention has excellent compatibility. Yu Qing Kailing Quan Fang Group. It shows that while the pharmaceutical composition of the present invention has the advantage of attenuating toxicity compared with Qingkailing Quanfang, the synergistic effect is also more obvious.

Based on the above two analysis results, the optimal range of compatibility ratio of the pharmaceutical composition of the present invention is determined.

Experimental example 4 Effect of the pharmaceutical composition of the present invention on the expression of cell adhesion molecules in the brain tissue of MCAO rats

1. Effect of the pharmaceutical composition of the present invention on the expression of ICAM-1 in brain tissue at different periods of cerebral ischemia

As shown in Table 1, the expression level of ICAM-1 in the brain tissue of rats in the model group was higher than that in the sham operation group at two periods after cerebral ischemia (P<0.05 or 0.01). After 12 hours of ischemia, the level of ICAM-1 in the pharmaceutical composition group of the present invention was significantly lower than that of the model group (P<0.01). There is no significant difference in the expression level of ICAM-1 of Lingling and the pharmaceutical composition of the present invention compared with the model group. It shows that the pharmaceutical composition of the present invention can inhibit the high expression of ICAM-1 in the very early stage of ischemia.

Table 1 ICAM-1 expression level in brain tissue homogenate supernatant at different periods of ischemia (X±SD ng/ml) group no Ischemia 12h Ischemia 24h mock surgical group 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 ^△△ Qing Kai Ling 10 0.779±0.113 1.62±0.309 ^△△

Note: Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01

Compared with the model group *P＜0.05 **P＜0.01

2. Effect of the pharmaceutical composition of the present invention on the expression of E-selectin in brain tissue at different periods of cerebral ischemia

As shown in Table 2, at two periods after cerebral ischemia, the expression level of E-selectin in the brain tissue of rats in the model group was higher than that in the sham operation group (P<0.01 or 0.05). After 12 hours of ischemia, the level of E-selectin in the brain tissue of the pharmaceutical composition group of the present invention was lower than that of the model group (P<0.01), and the Qingkailing group did not show a significant inhibitory expression effect; at 24 hours of ischemia, compared with the model group, the E-selectin level of the present invention Both the pharmaceutical composition group and the Qingkailing group showed significant expression suppression effects (P<0.01 or 0.05), and the effect of the former was better than that of the latter.

Table 2 E-selectin expression level in brain tissue homogenate supernatant at different periods of ischemia (X±SD ng/ml) group no Ischemia 12h Ischemia 24h Sham operation group, model group, pharmaceutical composition group of the present invention, Qingkailing group 10101010 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 ^*

Note: Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01

Compared with the model group *P＜0.05 **P＜0.01

Experimental Example 5 Effect of the pharmaceutical composition of the present invention on the vascular regulation function of MCAO rats

1. The effect of the pharmaceutical composition of the present invention on the plasma ET-1 content in different periods of cerebral ischemia.

As shown in Table 3, at 12h and 24h of cerebral ischemia, the plasma ET-1 content of the model group increased significantly, compared with the sham operation group, P<0.05; Compared with the model group, P>0.05. At 24 hours of cerebral ischemia, the plasma ET-1 content of the pharmaceutical composition group of the present invention was significantly lower than that of the model group, P<0.01, but Qingkailing still had no effective effect at this time.

Table 3 Plasma ET-1 content in different periods of cerebral ischemia (X±SD pg/ml) group no Ischemia 12h Ischemia 24h Sham operation group, model group, pharmaceutical composition group of the present invention, Qingkailing group 10101010 142.26±18.61159.06±7.66 ^△ 162.69±24.48 ^△ 158.35±6.32 ^△ 150.46±22.90174.30±25.33 ^△ 129.74±21.02 ^**△ 179.20±19.15 ^△

Note: Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01

Compared with the model group *P＜0.05 **P＜0.01

2. Effect of the pharmaceutical composition of the present invention on the ratio of plasma TXB ₂ /6-keto-PGF ₁ α in different periods of cerebral ischemia

As shown in Table 4, at 12h and 24h of cerebral ischemia, the plasma TXB ₂ /6-keto-PGF ₁ α ratio of the model group was significantly higher than that of the sham operation group P<0.01 or 0.05; The ratio of TXB ₂ /6-keto-PGF ₁ α in the Qingkailing group was significantly lower than that in the model group P<0.01; after 24 hours of ischemia, the ratio of plasma TXB ₂ /6-keto-PGF ₁ α in the pharmaceutical composition group of the present invention was still significantly lower In the model group, P<0.01, while the ratio of plasma TXB ₂ /6-keto-PGF ₁ α in the Qingkailing group was significantly higher than that in the model group, P<0.05, and there was no statistically significant difference with the model group.

Table 4 Ratio of plasma TXB ₂ /6-keto-PGF ₁ α in different periods of cerebral ischemia (X±SD) group no Ischemia 12h Ischemia 24h Sham operation group, model group, pharmaceutical composition group of the present invention, Qingkailing group 10101010 0.350±0.0910.709±0.168 ^△△ 0.331±0.178 ^** 0.352±0.087 ^** 0.366±0.0830.526±0.204 ^△ 0.251±0.092 ^** 0.704±0.326 ^△

Note: Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01

Compared with the model group *P＜0.05 **P＜0.01

Experimental Example 6 Effect of the pharmaceutical composition of the present invention on the content of inflammatory cytokines in the brain tissue of MCAO rats

1. Effect of the pharmaceutical composition of the present invention on TNF-α content in ischemic brain tissue

Table 5: TNF-α protein content in brain tissue homogenate at different periods of cerebral ischemia (X±SD pg/ml) group no Ischemia 12h no Ischemia 24h Sham operation group, model group, pharmaceutical composition group of the present invention, Qingkailing group 89109 15.241±1.18017.459±0.261 ^△ 17.747±3.114 ^△ 18.762±0.939 ^△△ 10101010 26.624±1.82328.324±1.292 ^Δ 24.715±2.843 ^** 25.725±2.841 ^*

Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01

Compared with the model group *P＜0.05 **P＜0.01

The results showed that: at two time points of cerebral ischemia 12h and 24h, the TNF-α protein content in the model group was significantly higher than that in the sham operation group p<0.05, indicating that ischemic stimulation promoted the synthesis and secretion of TNF-α. There was no significant difference between each administration group and the model group after ischemia for 12 hours. At 24 hours of ischemia, the two administration groups were significantly lower than the model group p<0.01 or 0.05.

2. Effect of the pharmaceutical composition of the present invention on the content of IL-1β in ischemic brain tissue

Table 6 IL-1β protein content of brain tissue homogenate at different periods of cerebral ischemia (X±SD pg/ml) group no Ischemia 12h no ischemia 4h Sham operation group, model group, pharmaceutical composition group of the present invention, geniposide group 89109 0.121±0.0210.143±0.018 ^△ 0.090±0.048 ^** 0.081±0.040 ^** 10101010 0.191±0.0230.221±0.042 ^△ 0.150±0.047 ^** 0.149±0.040 ^**

Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01,

Compared with the model group *P＜0.05 **P＜0.01

Table 6 shows that at 12h and 24h of ischemia, the IL-1β content in the brain tissue of the model group was significantly higher than that of the sham operation group, p<0.05, indicating that ischemia stimulates the synthesis and secretion of IL-1β. However, the two medication groups all showed a good inhibitory effect on this change, and compared with the model group, there was a significant statistical difference p<0.01.

Experimental Example 7 The protective effect of the pharmaceutical composition of the present invention on the damage of blood vessels and neurons in the brain tissue of MCAO rats

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

Table 7 Plasma vWF content in different periods of cerebral ischemia (X±SD unit) group no Ischemia 12h no Ischemia 24h Sham operation group, model group, pharmaceutical composition group of the present invention, Qingkailing group 89109 0.028±0.0080.062±0.017 ^△△ 0.055±0.023 ^△△ 0.068±0.021 ^△△ 10101010 0.024±0.0070.044±0.013 ^△△ 0.029±0.010 ^** 0.039±0.009 ^△

Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01,

Compared with the model group *P＜0.05 **P＜0.01

Table 7 shows that at 12h and 24h of ischemia, the plasma vWF content of the model group was significantly higher than that of the sham operation group, p<0.01. That ischemia caused significant damage to vascular endothelial cells. After 12 hours of ischemia, there was no significant statistical difference between the two drug groups and the model group. After 24 hours of ischemia, the pharmaceutical composition group of the present invention was significantly lower than the model group, p<0.01, but there was no significant statistical difference between the Qingkailing group and the model group.

2, the influence of pharmaceutical composition of the present invention on serum NSE content

Table 8 Serum NSE content in different periods of cerebral ischemia (X±SD ng/ml) group no Ischemia 12h no Ischemia 24h Sham operation group, model group, pharmaceutical composition group of the present invention, Qingkailing group 6668 3.33±0.698.72±4.79 ^△ 4.67±1.98 ^* 6.32±2.46 ^△ 76910 3.44±0.705.85±2.34 ^△ 4.58±1.19 ^△ 5.34±2.46 ^△

Compared with the sham operation group, ^△ P<0.05 ^△△ P<0.01, compared with the model group *P<0.05 **P<0.01

Table 8 shows that after 12h and 24h of cerebral ischemia, the serum NSE content of the model group increased, which was significantly different from that of the sham operation group, P<0.05. At 12 hours of ischemia, compared with the model group, the serum NSE content of the pharmaceutical composition group of the present invention was significantly reduced, P<0.05. However, the serum NSE content of the Qingkailing group was significantly higher than that of the sham operation group (P<0.05), and there was no significant decrease compared with the model group. After 24 hours of ischemia, compared with the model group, the serum NSE content of the two drug groups did not decrease significantly.

The preparation of embodiment 1 capsule of the present invention

Cholic acid 400g Baicalin 450g Gardenia 2300g Mother-of-pearl powder 4200g

Prepare capsules according to conventional methods.

The preparation of embodiment 2 sprays of the present invention

Cholic acid 740g Baicalin 500g Gardenia 2800g Mother-of-pearl powder 4500g

Prepare a spray according to a conventional method.

The preparation of embodiment 3 dropping pills of the present invention

Cholic acid 800g Baicalin 560g Gardenia 3000g Mother-of-pearl powder 5500g

Prepare drop pills according to conventional methods.

Embodiment 4 Preparation of oral liquid preparation of the present invention

Cholic acid 650g Baicalin 550g Gardenia 2500g Mother-of-pearl powder 5000g

Prepare oral liquid preparations according to conventional methods.

Embodiment 5 Preparation of tablet for injection of the present invention

Cholic acid 700g Baicalin 530g Gardenia 2600g Mother-of-pearl powder 4800g

Prepare tablets for injection according to conventional methods.

The preparation of embodiment 6 injections of the present invention

Taurocholic acid 350g Isodeoxycholic acid 350g Baicalin 550g

Gardenia 2500g Mother of Pearl Powder 5000g

Injections are made by conventional methods.

Embodiment 7 Preparation of freeze-dried powder injection of the present invention

Taurocholic Acid 480g Isodeoxycholic Acid 400g Baicalin 450g

Gardenia 3300g Mother of Pearl Powder 4300g

It is prepared into freeze-dried powder injection according to conventional methods.

The preparation of embodiment 8 granules of the present invention

Taurocholic Acid 370g

Gardenia 2400g Mother of Pearl Powder 5600g

Prepare granules according to conventional methods.

The preparation of embodiment 9 examples tablet of the present invention

Taurocholic Acid 150g Isodeoxycholic Acid 150g Baicalin 510g

Gardenia 2800g Mother of Pearl Powder 4500g

Prepare tablets according to conventional methods.

The preparation of embodiment 10 suspension of the present invention

Taurocholic Acid 350g Isodeoxycholic Acid 350g Baicalin 500g

Gardenia 2700g Mother of Pearl Powder 5000g

Suspensions are prepared by conventional methods.

The preparation of embodiment 11 injection of the present invention

A. Take 350g taurocholic acid, 390g isodeoxycholic acid, and 500g baicalin

Gardenia 2800g Mother of Pearl Powder 4500g

Prepare 100 liters of injection as follows.

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 8 times the amount of 70% ethanol, collect the percolation liquid, recover the 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 , 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, elute with distilled water until the eluent is colorless, and then use 10 Doubling the amount of 70% ethanol for elution, collecting the eluate, recovering the ethanol under reduced pressure and concentrating, drying 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 Refrigerate at ℃ 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, then weigh the mother-of-pearl powder and slowly add it under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling slightly for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution; after cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, mix them evenly, adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve it completely, add tauric acid powder to adjust the pH to 9 to dissolve it completely; The mixed extract is mixed in the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 suction filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

Adjust the pH of the mixed medicinal liquid to 7.2, add activated carbon (0.3) 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 balls to prepare for potting; Prepare injection solution according to the conventional method of pharmacy, 10ml/branch, 0.09g/ml, intravenous injection 20-40ml/day.

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

A. Take 390g of taurocholic acid, 410g of isodeoxycholic acid, and 560g of baicalin

Gardenia 3000g Mother of Pearl Powder 5500g

Make 100 liters of large injection infusion according to the following method.

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 6 times the amount of 70% ethanol, collect the percolation liquid, recycle the ethanol, concentrate to a clear paste with a relative density of 1.10 at 60°C, add 5 times the 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 Doubling the amount of 70% ethanol for elution, collecting the eluate, recovering the ethanol under reduced pressure and concentrating, drying 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 Refrigerate at ℃ for 36 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; then weigh the mother-of-pearl powder and add it slowly under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution. After cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, and mix them evenly; adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve all, at this time the pH is ~ 9, add taurocholic acid powder to adjust the pH to 9 Dissolve it completely; mix the mixed extract with the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 suction filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

Adjust the pH of the mixed medicinal liquid to 7.2, add activated carbon (0.3) 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 balls to prepare for potting; Take the total mixed medicinal solution, and prepare it into a large infusion solution according to the conventional method.

The preparation of embodiment 13 capsules of the present invention

A. Take 200g of taurocholic acid, 200g of isodeoxycholic acid, and 450g of baicalin

Gardenia 2300g Mother of Pearl Powder 4200g

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 6 times the amount of 70% ethanol, collect the percolation liquid, recycle the ethanol, concentrate to a clear paste with a relative density of 1.10 at 60°C, add 5 times the 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 Doubling the amount of 70% ethanol for elution, collecting the eluate, recovering the ethanol under reduced pressure and concentrating, drying 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 Refrigerate at ℃ for 36 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; then weigh the mother-of-pearl powder and add it slowly under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution; after cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, and mix them evenly; adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve all, at this time the pH is ~ 9, add taurocholic acid powder to adjust the pH to 9 Dissolve it completely; mix the mixed extract with the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 suction filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

The total mixed medicinal solution is concentrated under reduced pressure, spray-dried into powder, filled into hollow capsules, and prepared into hard capsules.

The preparation of embodiment 14 sprays of the present invention

A. Take 350g taurocholic acid, 350g isodeoxycholic acid, and 530g baicalin

Gardenia 2600g Mother of Pearl Powder 4800g

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 10 times the amount of 70% ethanol, collect the percolation liquid, recover the ethanol, concentrate it to a clear paste with a relative density of 1.10 at 60°C, add 3 times the amount of water to dilute , stir well, refrigerate at 0-4°C for 60 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, elute with distilled water until the eluent is colorless, and then use 8 Doubling the amount of 70% ethanol for elution, collecting the eluate, recovering the ethanol under reduced pressure and concentrating, drying 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 Refrigerate at ℃ for 12 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; then weigh the mother-of-pearl powder and add it slowly under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution. After cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, and mix them evenly; adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve all, at this time the pH is ~ 9, add taurocholic acid powder to adjust the pH to 9 Dissolve it completely; mix the mixed extract with the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 suction filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

Adjust the pH of the mixed medicinal liquid to 7.2, add activated carbon (0.3) 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 balls to prepare for potting; Take the total mixed medicinal solution and prepare it into an injection; concentrate the injection under reduced pressure to 1/2 volume, and fill each 10ml into a micro-spray bottle; prepare 5000 sprays, and the daily dosage is 3-5ml.

The preparation of embodiment 15 dropping pills of the present invention

A. Take 300g of taurocholic acid, 350g of isodeoxycholic acid, and 550g of baicalin

Gardenia 2500g Mother of Pearl Powder 5000g

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 8 times the amount of 70% ethanol, collect the percolation liquid, recycle the ethanol, concentrate to a clear paste with a relative density of 1.10 at 60°C, add 4 times the amount of water 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 Eluted with 10 times the amount of 70% ethanol, collected the eluent, recovered the ethanol under reduced pressure, concentrated, dried, and obtained the semi-finished product of Gardenia; dissolved the semi-finished product of Gardenia with water for injection, filtered; adjusted the pH value to 6.0-7.0, 0- Refrigerate at 4°C 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; then weigh the mother-of-pearl powder and add it slowly under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution; after cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, and mix them evenly; adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve all, at this time the pH is ~ 9, add taurocholic acid powder to adjust the pH to 9 Dissolve it completely; mix the mixed extract with the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 suction filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

Adjust the pH of the mixed medicinal liquid to 7.2, add activated carbon (0.3) 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 balls to prepare for potting; Concentrate the total mixed medicinal solution under reduced pressure, and spray dry it into powder; dissolve the medicinal powder and matrix substance (the ratio is PEG4000: stearic acid: paraffin: medicinal powder = 3.60: 0.32: 0.08: 1.00) in a water bath, mix well, and use an inner diameter of 1.69 mm, 3.81mm outer diameter dropper at 75°C with a drop rate of about 48 grains/min, drop into the liquid paraffin cooling liquid from top to bottom, collect the dropping pills and absorb the cooling agent with filter paper; prepared into drops pill.

Embodiment 16 Preparation of freeze-dried powder injection of the present invention

A. Take 300g of taurocholic acid, 430g of isodeoxycholic acid, and 510g of baicalin

Gardenia 2800g Mother of Pearl Powder 4500g

Make freeze-dried powder injection according to the following method.

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 8 times the amount of 70% ethanol, collect the percolation liquid, recover the 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 , 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, elute with distilled water until the eluent is colorless, and then use 10 Doubling the amount of 70% ethanol for elution, collecting the eluate, recovering the ethanol under reduced pressure and concentrating, drying 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 Refrigerate at ℃ 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, then weigh the mother-of-pearl powder and slowly add it under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling slightly for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution; after cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, and mix them evenly; adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve all, at this time the pH is ~ 9, add taurocholic acid powder to adjust the pH to 9 Dissolve it completely; mix the mixed extract with the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 suction filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

Adjust the pH of the mixed medicinal liquid to 7.2, add activated carbon (0.3) to boil in a water bath, filter while it is hot, seal and refrigerate the filtrate for 3-4 days, pulp and filter, adjust the pH to 7.2, and freeze-dry to make a freeze-dried powder.

Embodiment 17 Preparation of oral liquid preparation of the present invention

A. Take 200g taurocholic acid, 400g isodeoxycholic acid, and 500g baicalin

Gardenia 2700g Mother of Pearl Powder 5000g

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 6 times the amount of 70% ethanol, collect the percolation liquid, recover the ethanol, concentrate to a clear paste with a relative density of 1.10 at 60°C, and dilute with 5 times the amount of water , 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, elute with distilled water until the eluent is colorless, and then use 10 Elute with 70% 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, filter; adjust the pH value to 6.0-7.0, 0-4 Refrigerate at ℃ for 36 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; then weigh the mother-of-pearl powder and add it slowly under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling slightly for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution; after cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, and mix them evenly; adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve all, at this time the pH is ~ 9, add taurocholic acid powder to adjust the pH to 9 Dissolve it completely; mix the mixed extract with the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

Adjust the pH of the mixed medicinal liquid to 7.2, add activated carbon (0.3) to boil in a water bath, filter while it is hot, seal and refrigerate the filtrate for 3-4 days, filter the pulp, adjust the pH to 7.2, and use G4 filter balls to prepare for potting; The total mixed medicinal solution is prepared into an oral liquid preparation according to a conventional method.

The preparation of embodiment 18 injection tablet of the present invention

A. Take 320g taurocholic acid 480g isodeoxycholic acid 470g baicalin

Gardenia 3000g Mother of Pearl Powder 4000g

B. Preparation of gardenia extract and mother-of-pearl powder hydrolyzate

Take the prescribed amount of gardenia, crush it into coarse powder, percolate with 8 times the amount of 70% ethanol, collect the percolation liquid, recover the 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 , 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, elute with distilled water until the eluent is colorless, and then use 10 Elute with 70% 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, filter; adjust the pH value to 6.0-7.0, 0-4 Refrigerate at ℃ 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; then weigh the mother-of-pearl powder and add it slowly under constant stirring (note: keep stirring so that the liquid medicine does not overflow), heat to boiling and keep boiling for 6-7 hours. (constantly adding boiled deionized water) to filter, and the filter residue was washed twice with a small amount of hot water to obtain an orange-red clear solution; after cooling, white needle-like crystals were precipitated, and then filtered to remove the crystals to obtain an orange-red clear solution; Concentrate the clarified hydrolyzate to only 2-3 times the amount of the total amount of raw materials, cool it down, add ethanol to adjust the alcohol content to 60% and refrigerate for 24 hours; filter with suction, recover the ethanol from the filtrate until it has no alcohol smell, take it out and refrigerate for use For dosing.

C. Dosing

1. Mixing of extraction liquid:

Separately filter the gardenia extract and mother-of-pearl powder hydrolyzate, and mix them evenly; adjust the pH to 7 with 10% NaOH solution, and filter with suction to obtain a mixed extract;

2. Dissolution of bile acid:

Measure 75% ethanol (about 60 times the amount of cholic acid) with 10% NaOH solution to adjust the pH to 11, add deoxycholic acid powder to dissolve all, at this time the pH is ~ 9, add taurocholic acid powder to adjust the pH to 9 Dissolve it completely; mix the mixed extract with the bile acid solution to obtain a mixed medicinal solution;

3. Alcohol treatment of mixed liquid medicine:

Measure the mixed medicinal solution and add 95% ethanol 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 suction filter, recover the ethanol until it has no alcohol smell, and place it at room temperature; add When fresh water for injection is 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, pulp and suction filter, and a clear liquid should be obtained;

4. Dissolution and mixing of baicalin:

Measure fresh deionized water about the total volume of the solution, adjust the pH to 8 with 10% NaOH solution, add baicalein fine powder, adjust the pH to 7 with 10% NaOH solution, and dissolve it completely; add the solution containing baicalin into the mixed medicinal solution to obtain the total mixed medicinal solution.

D. Treatment of the total mixed liquid

The whole mixture is concentrated and prepared into tablets for injection according to conventional methods.

Claims (28)

1, a kind of pharmaceutical composition is characterized in that it being to be made by following bulk drugs:

Cholic acid 3-9 weight portion baicalin 4-6 weight portion

Fructus Gardeniae 20-35 weight portion Concha Margaritifera powder 40-60 weight portion.

2, pharmaceutical composition as claimed in claim 1 is characterized in that it being to be made by following bulk drugs:

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.

3, pharmaceutical composition as claimed in claim 1 is characterized in that it being to be made by following bulk drugs:

Cholic acid 4 weight portion baicalins 4.5 weight portions

Fructus Gardeniae 23 weight portion Concha Margaritifera powders 42 weight portions.

4, pharmaceutical composition as claimed in claim 1 is characterized in that it being to be made by following bulk drugs:

Cholic acid 8 weight portion baicalins 5.6 weight portions

Fructus Gardeniae 30 weight portion Concha Margaritifera powders 55 weight portions.

5, pharmaceutical composition as claimed in claim 1 is characterized in that it being to be made by following bulk drugs:

Cholic acid 7.4 weight portion baicalins 5.0 weight portions

Fructus Gardeniae 28 weight portion Concha Margaritifera powders 45 weight portions.

6, pharmaceutical composition as claimed in claim 1 is characterized in that it being to be made by following bulk drugs:

Cholic acid 6.5 weight portion baicalins 5.5 weight portions

Fructus Gardeniae 25 weight portion Concha Margaritifera powders 50.0 weight portions.

7, pharmaceutical composition as claimed in claim 1 is characterized in that it being to be made by following bulk drugs:

Cholic acid 7 weight portion baicalins 5.3 weight portions

Fructus Gardeniae 26 weight portion Concha Margaritifera powders 48 weight portions.

8,, it is characterized in that described cholic acid is made up of in the ratio of 2-4: 2-4.5 taurine and hyodesoxycholic acid as described any one pharmaceutical composition of claim 1-7.

9, pharmaceutical composition as claimed in claim 8 is characterized in that described cholic acid is made up of in the ratio of 3.0-3.5: 3.5-3.9 taurine and hyodesoxycholic acid.

10, pharmaceutical composition as claimed in claim 8 is characterized in that described cholic acid is made up of in 2.2: 3.4 ratio taurine and hyodesoxycholic acid.

11, pharmaceutical composition as claimed in claim 8 is characterized in that described cholic acid is made up of in 3.9: 4.1 ratio taurine and hyodesoxycholic acid.

12, pharmaceutical composition as claimed in claim 8 is characterized in that described cholic acid is made up of in 3.2: 3.8 ratio taurine and hyodesoxycholic acid.

13,, it is characterized in that being prepared into according to a conventional method the dosage form of clinical acceptance: ejection preparation, tablet, capsule, spray, pill, granule, suspensoid, drop pill or oral liquid as claim 1,2,3,4,5,6,7,9,10,11 or 12 described a kind of pharmaceutical compositions.

14, pharmaceutical composition as claimed in claim 8 is characterized in that being prepared into according to a conventional method the dosage form of clinical acceptance: ejection preparation, tablet, capsule, spray, pill, granule, suspensoid, drop pill or oral liquid.

15, preparation of drug combination method as claimed in claim 8 is characterized in that this method comprises the steps:

A. get taurine, hyodesoxycholic acid, baicalin, Fructus Gardeniae, Concha Margaritifera powder;

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 ejection preparation, tablet, capsule, spray, pill, granule, suspensoid, drop pill or oral liquid routinely.

16, preparation of drug combination method as claimed in claim 15 is characterized in that wherein the preparation method of gardenia extract is:

Get Fructus Gardeniae, be ground into coarse powder, 60%～80% ethanol percolation with 6～10 times of amounts, collect percolate, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear paste, add 3～5 times of water gaging dilutions, stir evenly, 0-4 ℃ of cold preservation 36～60 hours filters, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, it is colourless to be eluted to eluent with distilled water earlier, 8～12 times of amounts of reuse, 60%～80% ethanol elution, collect 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 12～36 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.

17, preparation of drug combination method as claimed in claim 16 is characterized in that wherein the preparation method of gardenia extract is:

Get Fructus Gardeniae, 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.

18, as claim 15,16 or 17 described preparation of drug combination methods, it is characterized in that wherein the preparation method of Concha Margaritifera powder hydrolyzed solution is:

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.

19, as described any one preparation of drug combination method of claim 9-12, it is characterized in that this method comprises the steps:

A. get taurine, hyodesoxycholic acid, baicalin, Fructus Gardeniae, Concha Margaritifera powder;

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 ejection preparation, tablet, capsule, spray, pill, granule, suspensoid, drop pill or oral liquid routinely.

20, preparation of drug combination method as claimed in claim 19 is characterized in that wherein the preparation method of gardenia extract is:

Get Fructus Gardeniae, be ground into coarse powder, 60%～80% ethanol percolation with 6～10 times of amounts, collect percolate, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear paste, add 3～5 times of water gaging dilutions, stir evenly, 0-4 ℃ of cold preservation 36～60 hours filters, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear paste, last activated-charcoal column, it is colourless to be eluted to eluent with distilled water earlier, 8～12 times of amounts of reuse, 60%～80% ethanol elution, collect 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 12～36 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.

21, preparation of drug combination method as claimed in claim 20 is characterized in that wherein the preparation method of gardenia extract is:

Get Fructus Gardeniae, 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.

22, preparation of drug combination method as claimed in claim 19 is characterized in that wherein the preparation method of Concha Margaritifera powder hydrolyzed solution is:

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.

23, preparation of drug combination method as claimed in claim 8 is characterized in that this method comprises the steps:

A. get taurine, hyodesoxycholic acid, baicalin, Fructus Gardeniae, Concha Margaritifera powder;

B. get Fructus Gardeniae, 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; 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;

C. gardenia extract, Concha Margaritifera powder hydrolyzed solution are filtered respectively, mix homogeneously is transferred PH to 7 with 10%NaOH liquid, and sucking filtration gets mixed extract; Be taken as 75% ethanol of 60 times of cholic acid amounts, 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; 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; Get the fresh deionized water of total 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. transfer total admixing medical solutions PH to 7.2, add the activated carbon 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 usefulness with G4 filter bulb sucking filtration, makes ejection preparation.

24, as described any one preparation of drug combination method of claim 9-12, it is characterized in that this method comprises the steps:

A. get taurine, hyodesoxycholic acid, baicalin, Fructus Gardeniae, Concha Margaritifera powder;

B. get Fructus Gardeniae, 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; 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;

C. gardenia extract, Concha Margaritifera powder hydrolyzed solution are filtered respectively, mix homogeneously is transferred PH to 7 with 10%NaOH liquid, and sucking filtration gets mixed extract; Be taken as 75% ethanol of 60 times of cholic acid amounts, 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; 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; Get the fresh deionized water of total 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. transfer total admixing medical solutions PH to 7.2, add the activated carbon 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 usefulness with G4 filter bulb sucking filtration, makes ejection preparation.

25, as claim 1,2,3,4,5,6,7,9,10, the application of 11 or 12 described pharmaceutical compositions in the medicine of preparation treatment cerebral ischemia.

26, the application of pharmaceutical composition as claimed in claim 8 in the medicine of preparation treatment cerebral ischemia.

27, application as claimed in claim 25 is wherein treated cerebral ischemia and is meant and prevents the ICAM-1 high expressed, prevent cerebral tissue E-selectin to express, reduce blood plasma ET-1 content, reduce blood plasma TXB ₂/ 6-keto-PGF ₁α ratio, the synthetic and secretion, reduction plasma vWF content or the reduction NSE contents in serum that reduce the synthetic of TNF-α and secrete, prevent IL-1 β.

28, application as claimed in claim 26 is wherein treated cerebral ischemia and is meant and prevents the ICAM-1 high expressed, prevent cerebral tissue E-selectin to express, reduce blood plasma ET-1 content, reduce blood plasma TXB ₂/ 6-keto-PGF ₁α ratio, the synthetic and secretion, reduction plasma vWF content or the reduction NSE contents in serum that reduce the synthetic of TNF-α and secrete, prevent IL-1 β.