CN1853689A - Chinese medicinal preparation for treating heart cerebrovascular disease and making method thereof - Google Patents

Abstract

The invention relates to a traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases and a preparation method thereof, in particular to a traditional Chinese medicine preparation prepared from two traditional Chinese medicines of red peony root and gardenia jasminoides, and particularly to the preparation of effective parts of the two traditional Chinese medicines Technology, the present invention also relates to the prescription of injection, molding technology and clinical dosage form.

Description

A traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases and its preparation method

technical field

The invention belongs to the technical field of medicine, and in particular relates to a traditional Chinese medicine preparation prepared from two traditional Chinese medicinal materials of Radix Paeoniae Rubra and Fructus Gardeniae, in particular to the preparation process of the effective parts of the two traditional Chinese medicinal materials. The invention also relates to the prescription and molding of injections Technology and clinical dosage forms.

technical background

Ischemic stroke is a kind of cardiovascular and cerebrovascular diseases. Although there are many drugs for the treatment of cardiovascular and cerebrovascular diseases in the world at present, due to the drug resistance, dependence and high price of western medicines, and the development of traditional Chinese medicine formulations and drug effects is relatively lagging behind, it is still necessary to continuously develop new high-efficiency, low-toxicity, and low-cost drugs. Reasonable, easy-to-use therapeutic drugs to meet clinical needs.

At present, the traditional Chinese medicine preparations used clinically to treat cardiovascular and cerebrovascular diseases and ischemic stroke are mainly oral preparations. Although oral preparations have certain curative effects, due to the first-pass effect of the dosage form, the onset of effect is slow and the bioavailability is low. , and it is inconvenient for stroke patients to take it, so it is far from meeting the needs of clinical medication.

Radix Paeoniae Rubra is the dried root of Paeonia lactiflora Pall., Paeonia obovatamaxim. or Paeonia veitchii Lynch; it is bitter in nature, slightly cold, and returns to the Liver Meridian; effect. The main active ingredients in Radix Paeoniae Rubra are water-soluble monoterpene glycosides, among which paeoniflorin has the highest content, and others include paeoniflorin, oxidized paeoniflorin, benzoylpaeoniflorin, and paeoniflorin.

According to literature reports, the traditional preparation methods of total paeoniflorin include water-alcohol method and alcohol-water method (see: editor-in-chief Yin Jian. Modern Research and Clinical Application of Chinese Medicine, Beijing: People's Health Publishing House, 2000: 354), this method The content of total paeoniflorin obtained is relatively low; in recent years, many reports in the literature have adopted macroporous adsorption resin refined glycosides (referring to: Chinese herbal medicine, 29 (10): 664-667), and the content of total paeoniflorin obtained by this method is relatively low. High; but because the adsorption and elution effects of macroporous resins are affected by many factors, in the application of this method, various operating conditions can be optimized to further increase the content of total paeoniflorin.

Gardenia is the dry and mature fruit of Gardenia jasminoides Ellis (Gardenia jasminoides Ellis). It is bitter and cold in nature, and has the effects of purging fire and relieving restlessness, clearing heat and diuresis, cooling blood and detoxifying. The main active ingredients in gardenia for the treatment of cardiovascular and cerebrovascular diseases are iridoid glycosides, mainly including geniposide, didhydroxygeniposide, argioside, and shangenin.

It has been reported in the literature that the content of total glycosides of gardenia is significantly higher than that of the traditional hydroalcoholic method by using macroporous adsorption resin for refining (see: Chinese herbal medicine, 33(9):794-796). On this basis, various operating conditions are optimized to further increase the content of total geniposides.

The present invention is prepared from the intermediates prepared by extracting two traditional Chinese medicinal materials of Radix Paeoniae Rubra and Gardenia Fructus. The active ingredient of the preparation of the present invention is clear, the quality is controllable, and it is convenient to use. The medicine of the present invention can be used to treat cardiovascular and cerebrovascular diseases. Disease, ischemic stroke, rapid onset, definite curative effect, high bioavailability, no drug resistance and dependence. Judging from the Chinese patent medicines listed for the treatment of cardiovascular and cerebrovascular diseases and ischemic stroke, there are no injection dosage forms made with the above two kinds of Chinese herbal medicines as prescriptions.

Contents of the invention

The invention provides a traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases. The preparation is made of Chinese medicine Radix Paeoniae Rubra and Radix Astragali as raw materials, which are extracted and processed to make active medicinal ingredients, which are further formulated with pharmaceutically acceptable carriers.

Another object of the present invention is to provide a preparation process for the effective parts of two kinds of Chinese medicinal materials - Radix Paeoniae Rubra and Gardeniae.

The Chinese medicine preparation of the present invention is made from the following Chinese medicine raw materials in weight ratio,

1-3 parts of red peony, 1-3 parts of gardenia, preferably 1.25-1.75 parts of peony, 1.25-1.75 parts of astragalus, more preferably: 1.67 parts of peony and 1.33 parts of astragalus.

Among the above compositions, the parts are parts by weight, and the weight is calculated based on crude drugs. If each part is in kilograms, the above composition can be made into 3000 doses of pharmaceutical preparations. The 3000 doses refer to the finished pharmaceutical preparations made, such as made 3,000 capsules, 3,000 tablets, 3,000 grams of granules, 3,000 ml of oral liquid, 3,000 sticks or 3,000 ml of injections, etc.

The above components are proportioned by weight, and can be increased or decreased according to the corresponding proportion during production. For example, the unit of kilogram or ton can be used for large-scale production, and the unit of milligram can also be used for small-scale production. It can be increased or decreased, but the weight ratio of raw medicinal materials among the components remains unchanged.

The proportion of the above weight ratio is obtained through scientific screening. For special patients, such as severe or mild, obese or thin patients, the proportion of the composition can be adjusted accordingly, and the increase or decrease does not exceed 100%. Change.

The single traditional Chinese medicine in the above composition can also be replaced by an appropriate traditional Chinese medicine with the same medicinal properties, and the medicinal effect of the replaced traditional Chinese medicine preparation remains unchanged.

The traditional Chinese medicine preparation of the present invention is prepared by extracting or otherwise processing the traditional Chinese medicine raw material composed of the above formula to make a pharmaceutically active substance, and then using the substance as a raw material, adding a pharmaceutically acceptable carrier when necessary, according to the conventional formula of pharmacy technology made. The active substance can be obtained by extracting the raw materials of traditional Chinese medicine separately, or by extracting the raw materials of traditional Chinese medicine together, or by other methods, such as: crushing, pressing, calcining, grinding, sieving, percolation, extraction, water extraction, Obtained by alcohol extraction, ester extraction, ketone extraction, chromatography and other methods, these active substances can be in the form of extracts, dry extracts or liquid extracts, and different concentrations can be determined according to different needs of the preparation.

The pharmaceutical active substance in the traditional Chinese medicine preparation of the present invention may account for 0.1-99.9% by weight in the preparation, and the rest is a pharmaceutically acceptable carrier. The pharmaceutical preparation of the present invention exists in the form of a unit dosage, and the unit dosage form refers to the unit of the preparation, such as every piece of a tablet, every capsule of a capsule, every bottle of an oral liquid, every bag of granules, and every injection. wait.

The Chinese medicine preparation of the present invention can be any pharmaceutically acceptable dosage form, and these dosage forms include: tablet, sugar-coated tablet, film-coated tablet, enteric-coated tablet, capsule, hard capsule, soft capsule, oral liquid, Buccal preparations, granules, granules, pills, powders, ointments, elixirs, suspensions, powders, solutions, injections, suppositories, ointments, plasters, creams, sprays, drops, patches. The preparation of the present invention is preferably an injection, most preferably a freeze-dried powder injection.

The traditional Chinese medicine preparation of the present invention, its oral administration preparation can contain commonly used excipients, such as binding agent, filler, diluent, tabletting agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent , and the tablets can be coated if necessary.

Suitable fillers include cellulose, mannitol, lactose and other similar fillers. Suitable disintegrants include starch, polyvinylpyrrolidone and starch derivatives such as sodium starch glycolate. Suitable lubricants include, for example, magnesium stearate. Suitable pharmaceutically acceptable wetting agents include sodium lauryl sulfate.

Solid oral compositions can be prepared by common methods such as mixing, filling, and tabletting. Repeated mixing is performed to distribute the active material throughout those compositions where large amounts of fillers are used.

Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example sorbitol, syrup, methylcellulose, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats, Emulsifiers, such as lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (they may include edible oils), such as almond oil, fractionated coconut oil, oily esters of esters such as glycerin, propylene glycol, or ethanol; preservatives agents, such as methyl or propyl paraben, or sorbic acid, and, if desired, conventional flavoring or coloring agents.

For injections, liquid unit dosage forms are prepared containing the active material of the invention in association with a sterile carrier. Depending on the carrier and concentration, the compound can be suspended or dissolved. Solutions are usually prepared by dissolving the active substance in a carrier and filter sterilizing before filling into a suitable vial or ampoule, which is then sealed. Adjuvants such as a local anesthetic, preservatives and buffering agents can also be dissolved in the vehicle. To increase its stability, the composition can be frozen after filling vials and the water removed under vacuum.

The traditional Chinese medicine preparation of the present invention can optionally add a suitable pharmaceutically acceptable carrier when it is prepared into a medicament, and the pharmaceutically acceptable carrier is selected from: mannitol, sorbitol, sodium metabisulfite, sodium bisulfite, thio Sodium sulfate, cysteine hydrochloride, thioglycolic acid, methionine, vitamin C, disodium EDTA, calcium sodium EDTA, monovalent alkali metal carbonates, acetates, phosphates or their aqueous solutions, hydrochloric acid, acetic acid, sulfuric acid, Phosphoric acid, amino acid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivatives, cellulose and its derivatives , alginate, gelatin, polyvinylpyrrolidone, glycerin, Tween 80, agar, calcium carbonate, calcium bicarbonate, surfactant, polyethylene glycol, cyclodextrin, β-cyclodextrin, phospholipid materials, kaolin , talcum powder, calcium stearate, magnesium stearate, etc.

When the preparation of the present invention is used, the usage and dosage are determined according to the patient's condition, and it can be taken three times a day, 1-20 doses each time, such as: 1-20 bags or granules or tablets.

The traditional Chinese medicine preparation of the present invention preferably adopts the following method to prepare.

(1) Take Radix Paeoniae Rubra, heat and extract 2 to 3 times with water, each time for 1 to 3 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.10 to 1.20, add ethanol to make the alcohol content reach 60 to 80% , add 10-20% alkali to make the pH value of the solution reach 6-8, refrigerate for 12-48 hours, and filter; the filtrate recovers ethanol until it has no alcohol smell, dilutes with water to a relative density of 1.10-1.20, stirs well, filters, and the filtrate Put it on a macroporous adsorption resin chromatography column, wash it with water until the eluent has no reducing sugar reaction, then elute it with 20-80% ethanol, collect the ethanol eluate, recover the ethanol under reduced pressure, and dry it in vacuum to obtain the total Glycoside;

(2) Take Gardenia, heat and extract with water for 2 to 3 times, each time for 1 to 3 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.10 to 1.20, add ethanol to make the alcohol content reach 60 to 80% , refrigerated for 12-48 hours, filtered; the filtrate recovered ethanol until it had no alcohol smell, diluted with water to a relative density of 1.10-1.20, added dilute hydrochloric acid to make the pH of the solution reach 2-3, refrigerated for 12-48 hours, filtered; the filtrate was added 10-20% alkali to make the pH value of the solution reach 7, stir well, filter, put the filtrate on a macroporous adsorption resin chromatography column, wash with water until the eluate has no reducing sugar reaction, and then elute with 20-80% ethanol, Collect the ethanol eluate, recover the ethanol under reduced pressure, and dry it in vacuum to obtain total glycosides of gardenia;

(3) Take the products of (1)-(2) above, mix them, and make pharmaceutical preparations according to the conventional techniques of pharmacy. Preferably, it is made into an injection, and the preparation method is as follows: take the above-mentioned products (1) to (2), mix them, add an appropriate amount of water for injection to dissolve once, add pharmaceutical excipients, stir to make them fully dissolve, add activated carbon for injection to the solution, Boil for 10-15 minutes, filter while hot, pass the filtrate through a microfiltration membrane to obtain a microfiltrate; pass the microfiltrate through an ultrafiltration membrane with a molecular weight of 6,000 to 30,000 to obtain an ultrafiltrate, and make the filtrate into injections according to the conventional preparation process, including liquid injections , Powder for injection.

The traditional Chinese medicine preparation of the present invention contains 1-3 parts of total glucosides of paeony and 0.5-6 parts of total glycosides of gardenia. Among them, the most preferred one contains 1 part of paeoniflorin and 2 parts of geniposide;

The active ingredients paeoniflorin and geniposide in the traditional Chinese medicine injection of the present invention are detected by high performance liquid phase method.

The pharmaceutical excipients of the present invention are one or more of Tuwen 80, mannitol, lactose, glucose, PVP, and sodium chloride; the total glucosides of paeony composition is mainly detected with paeoniflorin , the content of paeoniflorin is higher than 30%; the total glycoside composition of gardenia japonica uses geniposide as the main detection index, and the content of geniposide is higher than 40%.

The total glucosides of paeony, wherein the content of total glucosides of paeony is 50-95%, and the content of paeoniflorin is 30-60%.

The total glycosides of genipo, wherein the content of total glycosides of genipo is 50-95%, and the content of geniposide is 40-80%.

Prescription Screening Test

1. In vitro test (determination of platelet aggregation rate in vitro in rabbits):

1.1 Test method:

Rabbits with big ears were anesthetized with local subcutaneous injection of procaine, blood was bled into 10ml plastic test tubes with stoppers through carotid artery catheterization, anticoagulated with sodium citrate 1∶9, centrifuged at 800r/min for 10min, and platelet-rich in each test tube was taken out Plasma (PRP), the remaining part of the blood was centrifuged at 3000r/min, the supernatant plasma (platelet-poor plasma, PPP) was taken, platelet activating factor (PAF, final concentration 0.37μg/ml) was used as the aggregation inducer, and 250μl PRP was added to the test Add 10ul of different concentrations of drugs to the tube at the same time, add 10ul of normal saline to the control PRP, incubate for 5 minutes, and then use LG-PABER platelet aggregation coagulation factor analyzer (produced by Beijing Shidi Scientific Instrument Co., Ltd.) to detect platelets for 1min and 5min and the maximum (Max) aggregation rate.

1.2 Test drugs

Drug 1 total erythroside composition (paeoniflorin content is 30-60%), hereinafter expressed in the amount of paeoniflorin

Drug 2 Total glycosides composition of geniposide (40-80% geniposide content), expressed in geniposide amount below

1.3 Uniform design experiment and results of paeoniflorin and geniposide compatibility research:

13.1 Determine the level of uniform design according to the dose-effect relationship curve of paeoniflorin and geniposide:

The five levels (concentrations) of the uniform design are as follows:

Paeoniflorin in Drug 1: IC ₁₀ =1.53mg/ml, IC ₂₀ =2.59mg/ml, IC ₃₀ =4.18mg/ml, IC ₄₀ =5.51mg/ml, IC ₅₀ =6.83mg/ml

Geniposide in drug 2: IC ₁₀ =0.84mg/ml, IC ₂₀ =3.46mg/ml, IC ₃₀ =6.08mg/ml, IC ₄₀ =8.69mg/ml, IC ₅₀ =11.31mg/ml

1.3.2 Compatibility research of paeoniflorin and geniposide:

Using the U ₅ (5 ² ) table, see Table 1 for the results.

Table 1 Compatibility uniform design test results of paeoniflorin and geniposide (X±SD) Experiment number Concentration of paeoniflorin (mg/ml) Concentration of geniposide (mg/ml) Number of cases Platelet maximum aggregation rate (%) Aggregation inhibition rate (%) Control group 12345 -1(6.83)2(5.51)3(4.18)4(2.59)5(1.53) -2(8.69)5(0.84)3(6.08)1(11.31)4(3.46) 644444 55.32±6.71 20.88±2.64 ^** 35.50±5.17 ^** 26.58±5.14 ^** 26.80±5.03 ^** 44.28±6.10 ^* -62.3035.8052.0051.6020.00

The above results were analyzed by Master software, and the compatibility ratio of paeoniflorin and geniposide was determined to be 1:2.

2. In vivo test (determination of pulmonary embolism time in mice):

2.1 Test method:

98 mice were randomly divided into 7 groups, 14 in each group, half male and half male, ie control group, five dosage groups of drug administration group, positive control drug aspirin 40mg/kg dosage group. Each of the above groups was given 0.1ml/10g tail vein injection once, and aspirin was given by intragastric administration for 3 consecutive days. One hour after the last gavage and 5 minutes after intravenous injection, 0.1ml of mixed thrombogenic agent (ADP 15 μg/mouse, adrenaline 10 μg/mouse) was injected from the tail vein of the mouse, and the animal immediately appeared shortness of breath and could not move. The time required for mice to recover autonomous activities after the injection was taken as the time of pulmonary embolism, and the data were subjected to t-test between groups.

2.2 In vivo study on the compatibility of paeoniflorin and geniposide:

2.2.1 Dosage:

According to the relevant literature, the doses of paeoniflorin and geniposide were selected for a uniform design test to verify the compatibility ratio of the in vitro test and preliminarily determine the dose for in vivo administration. The specific dose levels are as follows:

The dose of paeoniflorin in drug 1 (mg/kg): 8.0, 4.0, 2.0, 1.0, 0.5

Geniposide dosage in drug 2 (mg/kg): 16.0, 8.0, 4.0, 2.0, 1.0

2.2.2 Uniform design test results:

Using the U ₅ (5 ² ) table, see Table 2 for the results.

Table 2 In vivo test results of paeoniflorin and geniposide in uniform design (X±SD) Experiment number Paeoniflorin dosage (mg/kg) Geniposide dosage (mg/kg) Number of cases Pulmonary embolism time (s) Time shortening rate (%) Control group Aspirin 12345 --1(8.0)2(4.0)3(2.0)4(1.0)5(0.5) --2(8.0)5(1.0)3(4.0)1(16.0)4(2.0) 14141414141414 294.21±87.30200.50±69.67 ^{** 166.43±64.16** 191.29±50.27** 186.93±57.83** 178.00 ±} 26.70 ^** 210.14±54.93 ^** -31.8543.4334.9836.4739.5028.57

The above results were analyzed by Master software, and it was confirmed that when the compatibility ratio of paeoniflorin and geniposide was 1:2, the time of pulmonary embolism was the shortest, and the dose of paeoniflorin was 0.5 mg/kg, and the dose of geniposide was 1.0 mg/kg. kg, there is still a good therapeutic effect.

3. Conclusion:

According to the above test results in vivo and in vitro, the compatibility is determined as follows:

The active ingredient ratio is: paeoniflorin 1, geniposide 2;

The proportion of intermediates is: total glucosides of paeony 1~3, total glycosides of gardenia 0.5~6;

The ratio of medicinal materials is: Radix Paeoniae Rubra 1-3, Gardenia 1-3.

Main Pharmacodynamic Test

1. Materials and methods

Drugs: Paeoniae Rubra + Gardenia

Nimodipine preparation stock solution (manufactured by Bayer AG of Germany, repackaged by Bayer Healthcare Co., Ltd. Beijing, China, batch number: CANAL ₂ ), 10mg/50ml, administered by intraperitoneal injection, 0.002g/kg, once a day.

aspirin

2. Determination of pulmonary embolism time

100 mice were randomly divided into 5 groups, 20 in each group, half male and half male, i.e. control group, three dose groups of test drug (red peony + gardenia) 1-20mg/kg, positive control drug aspirin 40mg/kg kg group. Each of the above groups was given 0.1ml/10g tail vein injection once, and aspirin was given by intragastric administration for 3 consecutive days. One hour after the last gavage and 5 minutes after intravenous injection, 0.1ml of mixed thrombogenic agents (15 μg of ADP and 10 μg of epinephrine) were injected from the tail vein of the mice, and the animals immediately appeared shortness of breath and could not move. The time required for the mice to recover voluntary activities was regarded as the time of pulmonary embolism, and the data were subjected to t-test between groups.

3. Determination of platelet aggregation rate in rabbits in vitro

The big-eared rabbits were locally anesthetized with procaine, bled by carotid artery catheterization, anticoagulated with 3.8% sodium citrate 1:9, centrifuged at 800r/min for 10min, platelet-rich plasma (PRP) was collected, and the rest was Centrifuge at 3000r/min, take platelet-poor plasma (PPP), use ADP (final concentration 15μmol/L) and PAF ((final concentration 0.37μg/ml) as the aggregation inducer, add 10ul of drugs with different concentrations in each tube of 250μl PRP, control 10 ul of buffer solution was added to the PRP, incubated for 5 minutes, and then the platelet aggregation rate was detected with an LG-PABER platelet aggregation coagulation factor analyzer (produced by Beijing Shidi Scientific Instrument Co., Ltd.).

4. Focal cerebral ischemia model in rats

The rats were randomly divided into 6 groups, namely normal group, model group, nimodipine group, and 3 dose groups of test drug (Red Paeoniae Rubra + Gardenia). Animals in each group were given intraperitoneal injection once a day for 3 consecutive days. After the last administration, the rats were taken, anesthetized with 10% chloral hydrate (400 mg/kg, ip), and cut the skin perpendicular to the line at the midpoint of the line between the right external auditory canal and the outer canthus of the right eye in a lateral position. 2cm, cut the fascia, bluntly separate the muscles, and expose the zygomatic arch. Bite off the zygomatic arch with hemostatic forceps, spread the zygomatic arch and mandible, pull and fix it on the mouse board with a small retractor, expose most of the squamous bone, and then about A small cranial window with a diameter of about 2 mm was drilled at 2 to 4 mm with a desktop electric drill. At this time, a small straight blood vessel with few branches can be seen through the dura mater, which is the middle cerebral artery (MCA). It passes through the olfactory bundle almost vertically and goes upward. Puncture the dura mater with fine acupuncture needles to expose the middle artery. After confirmation, set the electric knife to the bipolar coagulation position, select the largest electric coagulation switch, and coagulate 1mm inside the olfactory bundle to the brain A section of the middle cerebral artery between the inferior veins is electrocoagulated for 4 to 6 seconds. During electrocoagulation, to avoid ischemia and damage to the brain tissue, wet cotton balls can be used to protect it. After the middle cerebral artery was blocked, a small piece of muscle tissue was lightly applied to the cranial window, and then the wound was sutured layer by layer. After the operation, return to the cage for feeding. The above processes were carried out at a constant room temperature (24-25°C) to facilitate the evaluation of cerebral ischemia. Observe the effect on behavioral scores 4h and 24h after operation. After 24 hours of modeling, the carotid artery was bled to test related indicators, and the rats were sacrificed immediately, and the brain tissue was taken out to observe the changes of cerebral infarction area and infarction rate.

5. Mouse auricle swelling model

Immediately after the tail vein administration, xylene was applied to the front and back sides of the right ear of the mouse, 0.1 ml per mouse, and the left ear was used as a control. After 1 hour, the animals were sacrificed, and round ears were punched at the same site with a 9mm diameter puncher, and weighed with an electronic balance (HANGPING JA1203, Shanghai Balance Instrument Factory). Calculate the mouse ear swelling rate according to the formula.

Swelling rate (100%) = (right ear weight - left ear weight) / left ear weight × 100%

Swelling inhibition rate%=(the swelling rate of the control group-the swelling rate of the administration group)/the swelling rate of the control group×100%

result

1. Effects on the time of pulmonary embolism: the combination of Radix Paeoniae Rubra and Fructus Gardeniae can significantly shorten the time of pulmonary embolism in mice, and promote the recovery of autonomous activities in mice. Compared with the model group, there are significant differences.

2. Effects on the platelet aggregation rate of rabbits in vitro: In vitro administration of red peony root and gardenia in combination with various doses has inhibitory effects on the platelet aggregation rate of rabbits induced by PAF and ADP, and the difference between the control group and the control group is highly significant sex.

3. Effects on MCAO infarct size and behavior: staining of brain tissue sections showed that the infarcted brain tissue of model mice was white (normal brain tissue was rose red) with clear boundaries, and the behavioral score of the MCAO group was significantly increased. The infarct area and infarct rate in each dose group combined with Radix Paeoniae Rubra and Gardeniae were significantly reduced, and the behavioral score (24h) was also significantly decreased, which was significantly different from that in the MCAO model group.

4. Effects on the mouse auricle swelling model: each administration group combined with Radix Paeoniae Rubra and Gardenia had the effect of inhibiting the swelling of the mouse auricle caused by xylene, and there were significant differences compared with the model group, suggesting that this The product has a good anti-inflammatory effect.

The present invention utilizes macroporous resin separation technology, adopts aqueous solution to load sample, washes with water and then fully elutes with 50-90% ethanol, eluent adopts decompression to recover ethanol, and vacuum-dries each composition as an intermediate, and further forms according to the conventional method Process made into injections. The method has a high extraction rate of the composition, and the low-temperature treatment avoids the destruction of the active ingredients; the whole process is simple to operate, uses an internationally common non-secondary pollution solvent, reduces follow-up processing work, and reduces production costs, and is very suitable for industrial production. A large number of studies have shown that D101, AB-8, D-3520, HPD-400, and HPD-100 macroporous resins have better adsorption and separation effects on the above three compositions.

The present invention has the following characteristics:

1. The medicine of the present invention is composed of Radix Paeoniae Rubra and Fructus Gardeniae. The main active ingredients of Radix Paeoniae Rubra for treating cardiovascular and cerebrovascular diseases are water-soluble monoterpene glycosides. It is a water-soluble iridoid glycoside compound. Therefore, the preparation method of the medicine of the present invention selects a feasible route suitable for industrial production according to the physical and chemical properties of the main active ingredients contained in each medicine, and the active ingredients of the separated composition and final preparation are clear and controllable in quality.

2. The prescription screening of the present invention has undergone a large number of in vitro and in vivo experimental studies, and the dosage and compatibility used are reasonable, and the curative effect is definite; at the same time, the present invention uses the dosage of medicinal materials scientifically, avoiding the waste of medicinal material resources.

3. The dosage forms of the present invention are injections, including liquid injections and injection powder injections, which can be directly used for intramuscular injection and intravenous infusion, so they have rapid onset and high bioavailability, and are especially suitable for cardiovascular and cerebrovascular diseases and ischemic stroke Treatment.

4. The safety and stability experiments of the medicines and compositions of the present invention show that the indicators such as clarity, hemolysis, color, pH value, heat source, and irritation all meet the relevant regulations of the 2005 edition of the Chinese Pharmacopoeia for injections.

5. The freeze-dried powder injection of the present invention not only has the characteristics of injections, but also has the characteristics of convenient storage and portability; because the Chinese medicine components are prone to changes in physical and chemical properties in the solution state, it is especially suitable for Chinese medicine injections.

6. The main pharmacodynamic tests of the medicine of the present invention show that it has definite curative effect in the treatment of cardiovascular and cerebrovascular diseases and ischemic stroke.

The clinical action characteristics of the medicine of the present invention are mainly aimed at the treatment of ischemic stroke patients with blood stasis syndrome and ischemic stroke with both blood stasis blocking collaterals and blood-heat toxin pathogenic cardiovascular and cerebrovascular diseases and ischemic stroke. , from the perspective of clinical conditions, it is more suitable for treating patients in the acute stage of stroke and patients with serious conditions, especially those with hot constitution or internal heat due to yin deficiency. However, it can also be used for patients in the recovery period of stroke who meet the dialectics of traditional Chinese medicine.

The process of the present invention can increase the content of active ingredients, remove a large amount of impurities, and greatly reduce the amount of clinical medication; at the same time, the reproducibility and stability are good, and the quality is easy to control; and the yield of raw materials is high, the cost is low, and it is suitable for industrial production; Secondary polluted solvents are in line with international standards.

Detailed ways

The present invention is further illustrated by the following examples, but not as a limitation of the present invention.

Example 1

Preparation of freeze-dried powder injection

(1) Get 1.67Kg of Radix Paeoniae Rubra, add 18 times the amount of water, heat and extract three times, each time for 2 hours, the extracts are combined, filtered, the filtrate is concentrated to a relative density of 1.17, and ethanol is added to make the alcohol content reach 75%. Add 10% alkali to make the pH value of the solution 8, refrigerate for 24 hours, and filter; recover ethanol from the filtrate until it has no alcohol smell, add water to dilute to a relative density of 1.10-1.20, stir well, filter, and put the filtrate on a macroporous adsorption resin chromatography column, Wash with water until the eluate has no reducing sugar reaction, then elute with 70% ethanol, collect the ethanol eluate, recover ethanol under reduced pressure, and dry in vacuum to obtain 40.2 g of total glucosides of paeony, including 13.0 g of paeoniflorin.

(2) Take 1.33Kg of gardenia, add 24 times of water, heat and extract three times, each time for 2 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.12, add ethanol to make the alcohol content reach 80%, refrigerate After 48 hours, filter; recover ethanol from the filtrate until it has no alcohol smell, add water to dilute to a relative density of 1.10-1.20, add dilute hydrochloric acid to make the pH of the solution reach 3, refrigerate for 48 hours, and filter; add 10% alkali to the filtrate to make the pH of the solution reach 7. Stir well, filter, put the filtrate on a macroporous adsorption resin chromatography column, wash it with water until the eluate has no reducing sugar reaction, then elute with 80% ethanol, collect the ethanol eluate, recover ethanol under reduced pressure, vacuum After drying, 74.0 g of geniposides were obtained, which contained 30.1 g of geniposide.

(3) Take one-third of each of the above-mentioned (1)-(2) compositions, add appropriate amount of water for injection to dissolve each time, add pharmaceutical excipients, stir to fully dissolve, add activated carbon for injection into the solution, and boil for 10-15 minutes , filtered while hot, and the filtrate was passed through a microfiltration membrane to obtain a microfiltrate; the microfiltrate was passed through an ultrafiltration membrane with a molecular weight of 10,000 to obtain 1000 ml of an ultrafiltrate, and the filtrate was made into a freeze-dried powder injection according to a conventional preparation process.

Example 2

Preparation of Liquid Injections

(1) Get 1.67Kg of Radix Paeoniae Rubra, add 18 times the amount of water, heat and extract three times, each time for 2 hours, the extracts are combined, filtered, the filtrate is concentrated to a relative density of 1.17, and ethanol is added to make the alcohol content reach 75%. Add 10% alkali to make the pH value of the solution 8, refrigerate for 24 hours, and filter; recover ethanol from the filtrate until it has no alcohol smell, add water to dilute to a relative density of 1.10-1.20, stir well, filter, and put the filtrate on a macroporous adsorption resin chromatography column, Wash with water until the eluate has no reducing sugar reaction, then elute with 70% ethanol, collect the ethanol eluate, recover ethanol under reduced pressure, and dry in vacuum to obtain 40.2 g of total glucosides of paeony and 13.0 g of paeoniflorin.

(2) Take 1.33Kg of gardenia, add 24 times of water, heat and extract three times, each time for 2 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.12, add ethanol to make the alcohol content reach 80%, refrigerate After 48 hours, filter; recover ethanol from the filtrate until it has no alcohol smell, add water to dilute to a relative density of 1.10-1.20, add dilute hydrochloric acid to make the pH of the solution reach 3, refrigerate for 48 hours, and filter; add 10% alkali to the filtrate to make the pH of the solution reach 7. Stir well, filter, put the filtrate on a macroporous adsorption resin chromatography column, wash it with water until the eluate has no reducing sugar reaction, then elute with 80% ethanol, collect the ethanol eluate, recover ethanol under reduced pressure, vacuum After drying, 74.0 g of geniposide composition and 30.1 g of geniposide were obtained.

(3) Take one-third of each of the above-mentioned (1)-(2) compositions, add appropriate amount of water for injection to dissolve each time, add pharmaceutical excipients, stir to fully dissolve, add activated carbon for injection into the solution, and boil for 10-15 minutes , filtered while hot, and the filtrate was passed through a microfiltration membrane to obtain a microfiltrate; the microfiltrate was passed through an ultrafiltration membrane with a molecular weight of 30000 to obtain 1000 ml of an ultrafiltrate, and the filtrate was made into an injection by a conventional preparation process.

Example 3

preparation of tablets,

Step 1 and Step 2 are as in Step (1) and Step (2) of Examples 1 and 2, and Step (3) is to take one-third of the above-mentioned (1)-(2) compositions and add pharmaceutical excipients , prepared into 1000 tablets according to the conventional technology of photographic agents, and can be coated if necessary.

Example 4

preparation of capsules,

Step 1 and Step 2 are as in Step (1) and Step (2) of Examples 1 and 2, and Step (3) is to take one-third of the above-mentioned (1)-(2) compositions and add pharmaceutical excipients , prepared into 1000 capsules according to conventional techniques for capsules.

Example 5

preparation of granules,

Step 1 and Step 2 are as in Step (1) and Step (2) of Examples 1 and 2, and Step (3) is to take one-third of the above-mentioned (1)-(2) compositions and add pharmaceutical excipients , prepared into 1000g of granules according to the conventional technique of granules.

Claims (10)

1. A traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases, characterized in that it is made of the following Chinese medicine raw materials in the following proportions: 1 to 3 parts of Radix Paeoniae Rubra and 1 to 3 parts of Gardenia. 2. The traditional Chinese medicine preparation according to claim 1, which is characterized in that it is made of the following traditional Chinese medicine raw materials, 1.25-1.75 parts of peony and 1.25-1.75 parts of astragalus. 3. The traditional Chinese medicine preparation according to claim 1, which is characterized in that it is made of the following traditional Chinese medicine raw materials, 1.67 parts of peony and 1.33 parts of astragalus. 4. The traditional Chinese medicine preparation of claim 1, characterized in that it is selected from the following preparation forms, tablets, capsules, oral liquids, buccal preparations, granules, pills, powders, ointments, pills, suspensions, powders , solution, injection, suppository, ointment, plaster, cream, spray, drop, patch or pill. 5. The traditional Chinese medicine preparation according to claim 4, which is an injection. 6. The traditional Chinese medicine preparation according to claim 5, characterized in that said injection is a powder for injection. 7. The preparation method of the traditional Chinese medicine preparation according to claim 1, characterized in that, through the following steps, the raw materials of the traditional Chinese medicine composed of the formula are extracted or processed into other active substances, and the substance is used as the raw material, and the medicine can be added when necessary. The accepted carriers are prepared into pharmaceutical preparations according to conventional techniques of pharmacy. 8. The preparation method according to claim 7, characterized in that the extraction or other processing methods are selected from crushing, pressing, calcining, grinding, sieving, percolation, extraction, water extraction, alcohol extraction, ester extraction, ketone extraction or chromatography. 9. The preparation method according to claim 8, characterized in that, through the following steps, (1) Take Radix Paeoniae Rubra, heat and extract 2 to 3 times with water, each time for 1 to 3 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.10 to 1.20, add ethanol to make the alcohol content reach 60 to 80% , add 10-20% alkali to make the pH value of the solution reach 6-8, refrigerate for 12-48 hours, and filter; the filtrate recovers ethanol until it has no alcohol smell, dilutes with water to a relative density of 1.10-1.20, stirs well, filters, and the filtrate Put it on a macroporous adsorption resin chromatography column, wash it with water until the eluent has no reducing sugar reaction, then elute it with 20-80% ethanol, collect the ethanol eluate, recover the ethanol under reduced pressure, and dry it in vacuum to obtain the total Glycoside; (2) Take Gardenia, heat and extract with water for 2 to 3 times, each time for 1 to 3 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.10 to 1.20, add ethanol to make the alcohol content reach 60 to 80% , refrigerated for 12-48 hours, filtered; the filtrate recovered ethanol until it had no alcohol smell, diluted with water to a relative density of 1.10-1.20, added dilute hydrochloric acid to make the pH of the solution reach 2-3, refrigerated for 12-48 hours, filtered; the filtrate was added 10-20% alkali to make the pH value of the solution reach 7, stir well, filter, put the filtrate on a macroporous adsorption resin chromatography column, wash with water until the eluate has no reducing sugar reaction, and then elute with 20-80% ethanol, Collect the ethanol eluate, recover the ethanol under reduced pressure, and dry it in vacuum to obtain total glycosides of gardenia; (3) Take the products of the above (1) and (2), mix them, and make pharmaceutical preparations according to the conventional techniques of pharmacy. 10. The preparation method according to claim 9, characterized in that, through the following steps, (1) Get 1.67Kg of Radix Paeoniae Rubra, add 18 times the amount of water, heat and extract three times, each time for 2 hours, the extracts are combined, filtered, the filtrate is concentrated to a relative density of 1.17, and ethanol is added to make the alcohol content reach 75%. Add 10% alkali to make the pH value of the solution 8, refrigerate for 24 hours, and filter; recover ethanol from the filtrate until it has no alcohol smell, add water to dilute to a relative density of 1.10-1.20, stir well, filter, and put the filtrate on a macroporous adsorption resin chromatography column, Wash with water until the eluate has no reducing sugar reaction, then elute with 70% ethanol, collect the ethanol eluate, recover ethanol under reduced pressure, and dry in vacuum to obtain 40.2 g of total glucosides of paeony composition and 13.0 g of paeoniflorin; (2) Take 1.33Kg of gardenia, add 24 times of water, heat and extract three times, each time for 2 hours, combine the extracts, filter, concentrate the filtrate to a relative density of 1.12, add ethanol to make the alcohol content reach 80%, refrigerate After 48 hours, filter; recover ethanol from the filtrate until it has no alcohol smell, add water to dilute to a relative density of 1.10-1.20, add dilute hydrochloric acid to make the pH of the solution reach 3, refrigerate for 48 hours, and filter; add 10% alkali to the filtrate to make the pH of the solution reach 7. Stir well, filter, put the filtrate on a macroporous adsorption resin chromatography column, wash it with water until the eluate has no reducing sugar reaction, then elute with 80% ethanol, collect the ethanol eluate, recover ethanol under reduced pressure, vacuum Dried to obtain 74.0 g of geniposide composition and 30.1 g of geniposide; (3) Take one-third of each of the above-mentioned (1)-(2) compositions, add appropriate amount of water for injection to dissolve each time, add pharmaceutical excipients, stir to fully dissolve, add activated carbon for injection into the solution, and boil for 10-15 minutes , filtered while hot, and the filtrate was passed through a microfiltration membrane to obtain a microfiltrate; the microfiltrate was passed through an ultrafiltration membrane with a molecular weight of 30000 to obtain 1000 ml of an ultrafiltrate, and the filtrate was made into an injection by a conventional preparation process.