CN100409890C - Pharmaceutical composition for treating and preventing thrombus and its preparation method - Google Patents
Pharmaceutical composition for treating and preventing thrombus and its preparation method Download PDFInfo
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- CN100409890C CN100409890C CNB200510101011XA CN200510101011A CN100409890C CN 100409890 C CN100409890 C CN 100409890C CN B200510101011X A CNB200510101011X A CN B200510101011XA CN 200510101011 A CN200510101011 A CN 200510101011A CN 100409890 C CN100409890 C CN 100409890C
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Abstract
The present invention relates to a thrombus elimination medicine composition for treating and preventing thrombosis and a preparation method of the thrombus elimination medicine composition. A leech extract product is mainly made by leech by adopting the methods of extraction, coagulation and chromatographic fractionation, and extract products of curcuma roots and chuanxiong rhizome are made by adopting the methods of extraction and the purification of macroporous resin and using the curcuma roots and chuanxiong rhizome. The two extract products are merged and are made into preparations, such as freeze-drying powder injection, injection, tablets, capsules or soft capsules, etc. according to various kinds of preparation conventional means. The thrombus elimination medicine composition made by the present invention has the advantages of powerful thrombolytic effect, definite curative effect on diseases, such as cerebral infarction, myocardial infarction, etc. high security, efficiency of the prevention of thrombus formation, definite curative effect, controllable quality and small toxic side effect.
Description
Technical field:
The thrombosis that the present invention relates to a kind of treatment and prevention thrombosis is separated pharmaceutical composition and preparation method thereof
Background technology:
Thrombotic disease is the disease of a class serious harm human health and life, it comprises many commonly encountered diseases and frequently-occurring diseases such as coronary heart disease, myocardial infarction, ischemia apoplexy, venous thrombosis and pulmonary thromboembolism, and its sickness rate, disability rate and mortality rate are all very high.China's Chinese medicine culture is of extensive knowledge and profound scholarship, and traditional prescription is the treasure-house of a new drug development especially, and therefrom the newtype drug of exploitation treatment and prevention thrombosis is significant.It is ancient prescription that thrombosis is separated, and is made up of Hirudo, Radix Curcumae and Rhizoma Chuanxiong three flavor medical materials, is used for removing blood stasis and stimulates the menstrual flow, and cures mainly hemiplegia disturbance of consciousness, hemiplegia, and facial hemiparalysis, dysphonia, light red tongue has ecchymosis, stringy and hesitant pulse.
Hirudo is used to cure the disease all suitable history at home and abroad, and China is stated from Shennong's Herbal the beginning, and Hirudo can be treated hyperlipemia, has antithrombotic to form and thrombolytic effect.Its effective ingredient is mainly a kind of anticoagulative substance, hirudin (Hirudin) by name " it can stop thrombin to hematoblastic effect, suppresses platelet and is subjected to the stimulation of thrombin and discharges, thereby delay or stop solidifying of blood ".The effect of blood vessel dilating is arranged simultaneously, and can alleviate arteriospasm, blood viscosity lowering, microcirculation improvement, hematocrit, plasma viscosity, whole blood contrast viscosity, erythrocyte electrophoretic time, fibrinogen content and ESR equation k value to the cerebral infarction person also have tangible reduction effect.
Rhizoma Chuanxiong is samphire Rhizoma Chuanxiong (Ligusticum wallichii Frannch.) rhizome.Be clinical conventional Chinese medicine, have blood-activating and qi-promoting, an analgesic effect of going to become silted up.Modern study shows that the main effective ingredient of Rhizoma Chuanxiong is ligustrazine and ferulic acid, and Rhizoma Chuanxiong all has the strong and weak effect that does not wait to multisystem, summarizes and gets up to mainly contain the following aspects: 1. coronary artery dilating, increase coronary flow; 2. blood vessel dilating brings high blood pressure down; 3. microcirculation improvement; 4. anticoagulant, antithrombotic form and dissolve established thrombosis; 5. blood lipid regulation and atherosclerosis.
If this prescription uses inconvenience with the medical material direct drug injection, and utilization ratio of drug is low, and curative effect can not guarantee that quality can not be controlled.Therefore be necessary this medicine to be developed with modern extraction separation method and preparation means.
Summary of the invention
One of purpose of the present invention provides the pharmaceutical composition that the thrombosis of a kind of treatment and prevention thrombosis is separated, another object of the present invention provides this preparation of drug combination method, and a further object of the invention provides the application of this pharmaceutical composition in preparation treatment and prevention thrombosis medicine.
The thrombosis of a kind of treatment and prevention thrombosis is separated pharmaceutical composition, is by mass ratio 1-3 part Hirudo, 1-3 part Radix Curcumae, and the extract that 2-4 part Rhizoma Chuanxiong is made and the pharmaceutic adjuvant of corresponding dosage form are formed.
Preferred mass is than 1.5 portions of Hirudos, and the extract that 2 portions of Radix Curcumaes, 3 parts of Rhizoma Chuanxiongs are made and the pharmaceutic adjuvant of corresponding dosage form are formed.
Can be by 1.5 portions of Hirudos of mass ratio, extract and lyophilizing adjuvant that 2 portions of Radix Curcumaes, 3 parts of Rhizoma Chuanxiongs are made be made freeze-dried powder injection.
Can be by 1.5 portions of Hirudos of mass ratio, extract and corresponding dosage form adjuvant that 2 portions of Radix Curcumaes, 3 parts of Rhizoma Chuanxiongs are made be made injection, tablet, capsule or soft capsule.
The pharmaceutical composition that a kind of thrombosis is separated, make according to the following steps:
1) Hirudo powder is broken into coarse powder, with twice of the ethanol extraction of 70-90%, each 5-8 doubly measures, merging extracted twice liquid filters, filtrate is separated with chromatographic column or molecular sieve resin, after eluent concentrates, with the ultrafiltration of ultrafiltration post, solution between the intercepting molecular weight 3000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract;
2) Radix Curcumae, Rhizoma Chuanxiong are beaten coarse powder, with 75% ethanol extraction three times, merge extractive liquid, adds the water water precipitating after concentrating, and cold preservation is 24 hours in the freezer, filter the back and go up macroporous adsorbent resin, eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract;
3) Hirudo extract and Radix Curcumae Rhizoma Chuanxiong extract are merged, make corresponding preparations by various preparation routine techniquess.
After Hirudo extract and the preparation of Radix Curcumae Rhizoma Chuanxiong extract, the operation of the 3rd step can be with after Hirudo extract and the merging of Radix Curcumae Rhizoma Chuanxiong extract, be dissolved in the water for injection, cold preservation is spent the night, and filters the back and adds a certain amount of sodium sulfite, and the active carbon that adds 0.05-0.15% boils decarburization in 20 minutes, filter, packing behind the fine straining, injection or transfusion are made in sterilization.
After Hirudo extract and the preparation of Radix Curcumae Rhizoma Chuanxiong extract, the 3rd goes on foot after Hirudo extract and the merging of Radix Curcumae Rhizoma Chuanxiong extract, makes capsule or soft capsule by common process in this field.
After Hirudo extract and the preparation of Radix Curcumae Rhizoma Chuanxiong extract, the 3rd goes on foot after Hirudo extract and the merging of Radix Curcumae Rhizoma Chuanxiong extract, makes tablet by common process in this field.
A kind of injection thrombosis is separated the preparation method of pharmaceutical composition, it is characterized in that finishing according to the following steps:
1) Hirudo powder of getting 1.5 deals is broken into coarse powder, with twice of the ethanol extraction of 70-90%, each 5-8 doubly measures, merge extracted twice liquid and filter, filtrate is used 75 ethanol elutions with chromatographic column or molecular sieve resin chromatography, after eluent concentrates, with the ultrafiltration of ultrafiltration post, the solution between the intercepting molecular weight 3000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract;
2) get the Radix Curcumae of 2 deals and the Rhizoma Chuanxiong of 3 deals and beat coarse powder, with 75% ethanol extraction three times, merge extractive liquid, adds the water water precipitating after concentrating, and cold preservation is 24 hours in the freezer, filter the back and go up macroporous adsorbent resin, eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract;
A) Hirudo extract and Radix Curcumae Rhizoma Chuanxiong extract are merged fully be dissolved in the water for injection, add injection lyophilizing adjuvant,, filter through decarburization, fine straining, packing is jumped a queue, lyophilizing, gland adds aluminium lid and promptly gets the injection thrombosis and separate lyophilized powder.
Preferably hold back the material of 5000-20000 molecular weight in the Hirudo extract preparation process.
In the preparation process of Radix Curcumae Rhizoma Chuanxiong extract can be but be not limited to ADS-5, AB-8 or ADS-7 macroporous adsorbent resin.
The lyophilizing adjuvant can be but not limit that to select mannitol for use be the skeleton proppant.
More preferably ADS-5 macroporous adsorbent resin in the preparation process of Radix Curcumae Rhizoma Chuanxiong extract.
Gained pharmaceutical composition of the present invention can be used to prepare the medicine of treatment and prevention thrombosis.
The prepared thrombosis of the present invention is separated the pharmaceutical composition feasible process, determined curative effect, and toxic and side effects is little, can further be developed as the medicine of novel therapeutic and prevention thrombosis.
Specific embodiment:
Embodiment 1: the preparation of Hirudo extract
Get the 15kg Hirudo powder and be broken into coarse powder, add 70% medicinal alcohol 50L, after 60 ℃ of warm macerating extract 1.5h, filter, medicinal residues add 70% medicinal alcohol 50L again, and 60 ℃ of warm macerating extract 1.5h, filter, and merge filtrate twice.The good macroporous adsorbent resin of pretreatment is used 75% ethanol elution on the filtrate, after eluent concentrates, with the ultrafiltration of ultrafiltration post, the solution between the intercepting molecular weight 3000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract, weigh 37.3g.Lot number: 040201
Embodiment 2: the preparation of Hirudo extract
Get the 15kg Hirudo powder and be broken into coarse powder, add 90% medicinal alcohol 50L, after 60 ℃ of warm macerating extract 1.5h, filter, medicinal residues add 90% medicinal alcohol 50L again, and 60 ℃ of warm macerating extract 1.5h, filter, and merge filtrate twice.The good chromatographic column of pretreatment is used 75% ethanol elution on the filtrate, after eluent concentrates, with the ultrafiltration of ultrafiltration post, the solution between the intercepting molecular weight 5000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract, weigh 35.2g.Lot number: 040211
The assay of hirudin:
Get Fibrinogen 0.25g, be dissolved in the Tris2HCl buffer (pH=8) of 50mL, 0.5% fibrinogen solution " the dilution thrombin is to 5NIH/mL " is got the 0.2mL fibrinogen solution, add 20 μ L Hirudo extract lysates (getting 040201 and 040211 each 2mg respectively dissolves and be dissolved in surely in the 10ml volumetric flask) respectively, and add thrombin to solidifying, get hirudin content=concentration of thrombin * thrombin consumption/Hirudo extract liquor capacity, the results are shown in Table 1:
Table 1: hirudin assay
| Sample | Sampling amount (mg) | Hirudin content (AT-U/g) |
| 040201 | 2.82 | 4200 |
| 040211 | 2.94 | 4850 |
Embodiment 3: the preparation of Radix Curcumae Rhizoma Chuanxiong extract
Get the Radix Curcumae of 20kg and the Rhizoma Chuanxiong of 30kg and beat coarse powder, with 75% ethanol extraction three times, each 300L, merge extractive liquid, is concentrated into relative density 1.18 (50 ℃), add 6 times of water gagings and fully stir, cold preservation is 24 hours in the freezer, filters the back and goes up ADS-5 macroporous adsorbent resin (Tianjin Ourui Biology Technology Co., Ltd.), eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract, weigh 88.7g.Lot number: 040220
Embodiment 4: the preparation of Radix Curcumae Rhizoma Chuanxiong extract
Get the Radix Curcumae of 20kg and the Rhizoma Chuanxiong of 30kg and beat coarse powder, with 75% ethanol extraction three times, each 500L, merge extractive liquid, is concentrated into relative density 1.18 (50 ℃), add 6 times of water gagings and fully stir, cold preservation is 24 hours in the freezer, filters the back and goes up AB-8 macroporous adsorbent resin (Tianjin Ourui Biology Technology Co., Ltd.), eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract, weigh 96.8g.Lot number: 040225
Embodiment 5: the injection thrombosis is separated the preparation of lyophilized powder
With 040201 Hirudo extract 0.37g, 040220 Radix Curcumae Rhizoma Chuanxiong extract 0.89g, uniform mixing fully is dissolved in the 80ml water for injection, and cold preservation was filtered in 24 hours, filtrate adds 8g mannitol, crosses 0.45 micron microporous filter membrane, adds the injection water and is settled to 100ml, solution is heated to about 80 ℃, adds the 0.05g active carbon, keeps temperature 15 minutes, filter, fine straining is sub-packed in every 2ml in the 7ml cillin bottle, jump a queue, lyophilizing, gland adds aluminium lid and promptly gets the injection thrombosis and separate lyophilized powder.Lot number: lyophilized powder 040301
Embodiment 6: thrombosis is separated the preparation of injection
With 040201 Hirudo extract 0.37g, 040220 Radix Curcumae Rhizoma Chuanxiong extract 0.89g, uniform mixing fully is dissolved in the 800ml water for injection, cold preservation was filtered in 24 hours, and filtrate adds the sodium sulfite of 0.5g, fully dissolving, after adding water for injection and boiling to the 1000ml, the active carbon that adds 0.5g boils decarburization in 20 minutes, filters, and is sub-packed in the 100ml bottle after 0.22 micron micropore filter filters, sterilized 20 minutes, and made transfusion for 115 ℃.Lot number: transfusion 40310
Embodiment 7: thrombosis is separated preparation of soft capsule
With 040211 Hirudo extract 0.35g, 040225 Radix Curcumae Rhizoma Chuanxiong extract 0.97g, behind the uniform mixing, add soybean oil 15g, Cera Flava 0.3g, mixing is made 50 of soft capsules, promptly.
Embodiment 8: thrombosis is separated capsular preparation
With 040211 Hirudo extract 0.35g, 040225 Radix Curcumae Rhizoma Chuanxiong extract 0.97g, behind the uniform mixing, add 10% dextrin, granulate, make 50 of capsules, promptly
Embodiment 9: thrombosis is separated the preparation of sheet
With 040211 Hirudo extract 0.35g, 040225 Radix Curcumae Rhizoma Chuanxiong extract 0.97g, add 7.5g microcrystalline Cellulose, 0.5g crospolyvinylpyrrolidone behind the uniform mixing, pulverize, ethanol is softening, granulates 50 ℃ of dryings behind 60 mesh sieves excessively, cross 60 mesh sieve granulate, add the 0.5g carboxymethyl starch sodium, 0.1g Aspartane, 1.5g magnesium stearate uniform mixing, be pressed into 50, promptly.
Embodiment 10, external Sanguis Leporis seu oryctolagi bolt dissolution experiment
The experiment medicine: freeze-dried powder of the present invention, freeze-dried powder solution preparation of the present invention: get (040301) 1 and 5 of lyophilized powder respectively, each adds the solution that normal saline 100ml is prepared into two kinds of variable concentrations.
Normal saline: 0.9% sodium chloride solution
Radix Salviae Miltiorrhizae Injection is got 10ml, adds normal saline 40ml and is prepared into injection of danshen solution.
Laboratory animal: 2 of 1.0-1.5kg new zealand rabbits
Experimental technique: get blood in rabbit jugular vein, get intravenous rabbit blood 30ml (from footpath 0.34cm) in polyvinyl chloride pipe, allow it solidify under the room temperature, release grumeleuse with syringe behind the 60min, it is standby to be cut into the 0.15cm3 size, prepares 12 altogether, makes the whole blood grumeleuse.Get intravenous rabbit blood 30ml, use the sodium citrate anticoagulant, with the centrifugal 10min of 800r/min, collect upper plasma and middle level platelet, add thrombin then, add the 0.1ml (dosage of thrombin of 10 μ/ml) by every milliliter of blood plasma, in plate, form grumeleuse behind the mixing, the fritter that is cut into 0.15cm3 is standby, prepares 12 altogether, makes the platelet-rich plasma grumeleuse.Getting concentration is the pure fiber egg original solution 2ml of 20mg/ml (not containing fibrinolysin and plasminogen), add the 2ml (thrombin of 10 μ/ml), be to form grumeleuse in the small test tube of 0.5cm from the footpath, grumeleuse is shaken out gently, the fritter that is cut into 0.15cm3 is standby, prepare 12 altogether, make pure fibrin clot.Rabbit whole blood grumeleuse, rabbit platelet-rich plasma grumeleuse and the pure fibrin clot that makes dropped into respectively in freeze-dried powder solution of the present invention, injection of danshen liquor positive control solution and the normal saline negative controls of high and low concentration (being 600 μ l solution), put and be incubated and observe its dissolving situation in 37 ℃ of thermostatted water solution, write down its complete dissolution time, each experiment repeats to do 3 times.
Experimental result sees Table 2:
Table 2: each organizes (x ± s) of the consoluet time ratio of thrombosis
Annotate: injection thrombosis EGCT of the present invention and Radix Salviae Miltiorrhizae matched group are relatively
*P＜0.01
Embodiment 11, utilize the thrombosis method to measure to the thrombotic influence of rat
Experiment medicine: freeze-dried powder of the present invention, freeze-dried powder solution preparation of the present invention: get lyophilized powder (040301)
1 and 5, each adds the solution that normal saline 100ml is prepared into two kinds of variable concentrations
Normal saline: 0.9% sodium chloride solution
Radix Salviae Miltiorrhizae Injection is got 2ml, adds normal saline 50ml and is prepared into injection of danshen solution.
40 of the SD rat of laboratory animal: body weight 300-400 gram, male and female half and half.Be divided into blank, Radix Salviae Miltiorrhizae Injection matched group, injection low dosage of the present invention, injection high dose four components of the present invention at random, 10 every group.
Experimental technique: with the rat administration, dosage, freeze-dried powder solution low dose group of the present invention, freeze-dried powder solution high dose group of the present invention, the Radix Salviae Miltiorrhizae Injection matched group is irritated stomach.Capacity normal saline such as blank group, the filling gastric solubleness is long-pending to be 6ml/kg, once a day, totally 5 days.Rat administration the 5th day, after 1 hour, animal is weighed after the last administration, iv 3% sodium pentobarbital 1ml/kg anesthesia.Dorsal position is fixed, get the neck median incision, separate trachea, separate the total A of bilateral neck, V wears 6cm5 0 silk thread in polyethylene tube, be full of polyethylene tube with heparin-saline solution, after an end of polyethylene tube inserts the left side external jugular vein, inject anticoagulant heparin accurately by polyethylene tube, again the polyethylene tube other end is inserted right carotid artery, open bulldog clamp, cause the A-V bypass, open blood flow 15min takes out silica gel tube, extract silk thread (containing thrombosis) out, by number placing the little culture dish of having weighed, analytical balance takes by weighing wet weight of thrombus, and gross weight deducts silk thread weight and promptly gets thrombus weight.
The results are shown in Table 3:
Table 3: the thrombus weight comparison of each administration group (x ± s)
Annotate: compare * * P＜0.01 with the normal saline matched group
Compare △ △ P＜0.01 with the Radix Salviae Miltiorrhizae Injection matched group
Embodiment 12, to rat prothrombin time (PT), activated partial thromboplastin time (APTT) and hemorheology influence
Experiment medicine: freeze-dried powder of the present invention, freeze-dried powder solution preparation of the present invention: get lyophilized powder (040301)
1 and 5, each adds the solution that normal saline 100ml is prepared into two kinds of variable concentrations
Normal saline: 0.9% sodium chloride solution
Radix Salviae Miltiorrhizae Injection is got 2ml, adds normal saline 50ml and is prepared into injection of danshen solution.
40 of the SD rat of laboratory animal: body weight 300-400 gram, male and female half and half.Be divided into blank, Radix Salviae Miltiorrhizae Injection matched group, injection low dosage of the present invention, injection high dose four components of the present invention at random, 10 every group.
Experimental technique: with the rat administration, dosage, freeze-dried powder solution low dose group of the present invention is pressed, freeze-dried powder solution high dose group of the present invention, injection of danshen solution group is irritated stomach.Capacity normal saline such as blank group, the filling gastric solubleness is long-pending to be 6ml/kg, once a day, totally 5 days.Rat administration the 5th day, after the last administration after half an hour an eye artery get blood, get blood 2ml and add in the test tube contain 3.8% structure rafter acid sodium aqueous solution 0.2ml, put upside down the test tube mixing, detect by ACL-200 (U.S.'s Beckman) type intelligence blood agglutometer again and measure PT, APTT.Other gets blood 2ml, adding the inwall that has prepared in advance has in the dry test-tube of heparin, put upside down mixing, application of sample uses adjustable sample injector, transfer to 800 μ l, with test glass and cut the blood plate test macro of packing into, contact specimen cup inner surface at the bottom of the sample injector suction nozzle, allow blood slowly slip into along interior device, pre-warm 3 minutes by hemorheology analytical tool LG-R-80 (Beijing generation Supreme Being scientific instrument company limited) row whole blood viscosity, plasma viscosity, erythrocyte aggregation index, erythrocyte deformability detection.
The results are shown in Table 4,5:
Table 4: each administration group to P of Rats T and APTT influence (x ± s)
Annotate: compare * * P＜0.01 with the normal saline matched group
Compare △ P＜0.05 with the Radix Salviae Miltiorrhizae Injection matched group
Table 4: each administration group is to hemorheology of rat index influence (x ± s)
Annotate: compare * * P＜0.01 * P＜0.05 with the normal saline matched group
Embodiment 13, to the influence (turbidimetry) of rat platelet aggregation
The experiment medicine: freeze-dried powder of the present invention, freeze-dried powder solution preparation of the present invention: get (040301) 1 and 5 of lyophilized powder respectively, each adds the solution that normal saline 100ml is prepared into two kinds of variable concentrations
Normal saline: 0.9%
Radix Salviae Miltiorrhizae Injection is got 4ml, adds normal saline 50ml and is prepared into injection of danshen solution.
40 of the SD rat of laboratory animal: body weight 300-400 gram, male and female half and half.Be divided into blank, Radix Salviae Miltiorrhizae Injection matched group, injection low dosage of the present invention, injection high dose four components of the present invention at random, 10 every group.
Experimental technique: with the rat administration, dosage, freeze-dried powder solution low dose group of the present invention, freeze-dried powder solution high dose group of the present invention, the Radix Salviae Miltiorrhizae Injection group is irritated stomach.Capacity normal saline such as blank group, the filling gastric solubleness is long-pending to be 6ml/kg, once a day, totally 7 days.Rat administration the 7th day, the big rathole artery of going after half an hour after the last administration is got blood, 0.13% structure edge acid sodium anticoagulant (blood and anticoagulant volumetric ratio 9: 1), blood is injected silication in vitro, cover the centrifugal mouth of pipe with plastic sheeting, centrifuge tube reversed make blood and the abundant mixing of anticoagulant for 3-4 time, inhale attachment removal at the residual blood of tube wall, and step up the tight centrifuge tube of lid lid with the filter paper of cleaning.With 1000r/min centrifugal 10 minutes, carefully take out upper plasma (PRP), will remain blood plasma centrifugal 20 minutes with 3000r/min, lower floor's transparency liquid is PPP.Platelet count among the counting PRP transfers to 200 * 109/L with PPP with the platelet among the PRP.The pipet of silicidation gets PPP and PRP450u1 inserts in the opacity tube respectively, earlier the transmittance of monitor is adjusted to 100 during mensuration with the PPP specimen, then the PRP specimen is put into and measured the hole, regulating transmittance is 10, and adds the stirring bar magnet, 37 ℃ of preheatings 3 minutes, open monitor, in PRP, add ADP induced platelet aggregating agent prepared therefrom, CHROND-Log platelet aggregation instrument (U.S.), calculate platelet maximum agglutination rate (%) by assembling curve, (Amax)=h1/h0 * 100%; Platelet aggregation inhibition rate, suppression ratio=[(normal saline Amax-medication Amax)/normal saline Amax] * 100%.
The results are shown in Table 6
Table 6: each administration group to rat platelet aggregation influence (x ± s)
Annotate: compare * * P＜0.01 with the normal saline matched group
Give with Radix Salviae Miltiorrhizae Injection and to compare △ △ P＜0.01
Embodiment 14: to the influence of the inductive thrombosis death of mice ADP
The experiment medicine: freeze-dried powder of the present invention, freeze-dried powder solution preparation of the present invention: get (040301) 1 and 5 of lyophilized powder, each adds the solution that normal saline 100ml is prepared into two kinds of variable concentrations
Normal saline: 0.9% sodium chloride solution
Radix Salviae Miltiorrhizae Injection is got 5ml, adds normal saline 50ml and is prepared into injection of danshen solution.Laboratory animal: 80 of BALB/c mouse, male and female half and half.Be divided into blank, Radix Salviae Miltiorrhizae Injection matched group, freeze-dried powder solution low dosage of the present invention, freeze-dried powder solution high dose four components of the present invention at random, 20 every group.
Experimental technique: with the mice administration, dosage, freeze-dried powder solution low dose group of the present invention, freeze-dried powder solution high dose group of the present invention, the Radix Salviae Miltiorrhizae Injection group is irritated stomach.Capacity normal saline such as blank group, the filling gastric solubleness is long-pending to be 6ml/kg, once a day, and continuous 7 days, 1h tail vein injection ADP48.0ml/kg after the last administration, record animal dead situation.
The results are shown in Table 7:
Table 7: the influence to the inductive thrombosis death of mice ADP of each administration group is compared
Claims (10)
1. the pharmaceutical composition for the treatment of and preventing thrombosis is by mass ratio 1-3 part Hirudo, 1-3 part Radix Curcumae, and the ethanol extraction that 2-4 part Rhizoma Chuanxiong is made and the pharmaceutic adjuvant of corresponding dosage form are formed.
2. pharmaceutical composition as claimed in claim 1 is characterized in that by 1.5 portions of Hirudos of mass ratio, and the ethanol extraction that 2 portions of Radix Curcumaes, 3 parts of Rhizoma Chuanxiongs are made and the pharmaceutic adjuvant of corresponding dosage form are formed.
3. pharmaceutical composition as claimed in claim 2 is characterized in that by 1.5 portions of Hirudos of mass ratio, 2 portions of Radix Curcumaes, 3 parts of ethanol extraction and lyophilizing adjuvant compositions that Rhizoma Chuanxiong is made.
4. the pharmaceutical composition of a kind of treatment as claimed in claim 1 and prevention thrombosis, make according to the following steps:
1) Hirudo powder is broken into coarse powder, with twice of the ethanol extraction of 70-90%, each 5-8 doubly measures, merging extracted twice liquid filters, filtrate is separated with chromatographic column or molecular sieve resin, after eluent concentrates, with the ultrafiltration of ultrafiltration post, solution between the intercepting molecular weight 3000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract;
2) Radix Curcumae, Rhizoma Chuanxiong are beaten coarse powder, with 75% ethanol extraction three times, merge extractive liquid, adds aqueous dispersion after concentrating, and cold preservation is 24 hours in the freezer, filter the back and go up macroporous adsorbent resin, eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract;
3) Hirudo extract and Radix Curcumae Rhizoma Chuanxiong extract are merged, make corresponding preparations by various preparation routine techniquess.
5. pharmaceutical composition as claimed in claim 4 is characterized in that making according to the following steps:
1) Hirudo powder is broken into coarse powder, with twice of the ethanol extraction of 70-90%, each 5-8 doubly measures, merging extracted twice liquid filters, filtrate after eluent concentrates, is used the ultrafiltration of ultrafiltration post with chromatographic column or molecular sieve resin chromatography, solution between the intercepting molecular weight 3000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract;
2) Radix Curcumae, Rhizoma Chuanxiong are beaten coarse powder, with 75% ethanol extraction three times, merge extractive liquid, adds the water water precipitating after concentrating, and cold preservation is 24 hours in the freezer, filter the back and go up macroporous adsorbent resin, eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract;
3) Hirudo extract and the merging of Radix Curcumae Rhizoma Chuanxiong extract fully are dissolved in water for injection
In, add injection lyophilizing adjuvant, make the injection thrombosis with routine techniques in this field and separate lyophilized powder.
6. pharmaceutical composition as claimed in claim 4 is characterized in that making according to the following steps:
1) Hirudo powder is broken into coarse powder, with twice of the ethanol extraction of 70-90%, each 5-8 doubly measures, merging extracted twice liquid filters, filtrate after eluent concentrates, is used the ultrafiltration of ultrafiltration post with chromatographic column or molecular sieve resin chromatography, solution between the intercepting molecular weight 3000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract;
2) Radix Curcumae, Rhizoma Chuanxiong are beaten coarse powder, and with 75% ethanol extraction three times, merge extractive liquid, adds the water water precipitating after concentrating, and cold preservation is 24 hours in the freezer, filter the back and go up macroporous adsorbent resin, eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract;
3) Hirudo extract and Radix Curcumae Rhizoma Chuanxiong extract are merged, be dissolved in the water for injection, cold preservation is spent the night, and filters the back and adds a certain amount of sodium sulfite, the active carbon that adds 0.05-0.15% boils decarburization in 20 minutes, makes injection or transfusion by common process in this field.
7. the preparation method of a medicinal composition for injections is characterized in that finishing according to the following steps:
1) Hirudo powder of getting 1.5 deals is broken into coarse powder, with twice of the ethanol extraction of 70-90%, each 5-8 doubly measures, merging extracted twice liquid filters, filtrate after eluent concentrates, is used the ultrafiltration of ultrafiltration post with chromatographic column or molecular sieve resin chromatography, solution between the intercepting molecular weight 3000-20000, the cold drying of ultrafiltrate concentrating under reduced pressure gets Hirudo extract;
2) get the Radix Curcumae of 2 deals and the Rhizoma Chuanxiong of 3 deals and beat coarse powder, with 75% ethanol extraction three times, merge extractive liquid, adds the water water precipitating after concentrating, and cold preservation is 24 hours in the freezer, filter the back and go up macroporous adsorbent resin, eluent concentrate dry the Radix Curcumae Rhizoma Chuanxiong extract;
3) Hirudo extract and Radix Curcumae Rhizoma Chuanxiong extract are merged fully be dissolved in the water for injection, add injection lyophilizing adjuvant,, filter through decarburization, fine straining, packing is jumped a queue, lyophilizing, gland adds aluminium lid and promptly gets the injection thrombosis and separate lyophilized powder.
8. the preparation method of medicinal composition for injections as claimed in claim 7, what it is characterized in that holding back in the Hirudo extract preparation process is the material of 5000-20000 molecular weight.
9. as the preparation method of claim 7 or 8 described medicinal composition for injections, it is characterized in that resin uses ADS-5, AB-8 or ADS-7 macroporous adsorbent resin in the preparation process of Radix Curcumae Rhizoma Chuanxiong extract.
10. the application of pharmaceutical composition as claimed in claim 5 in preparation treatment and prevention thrombosis medicine.
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| CN101412726B (en) * | 2007-10-15 | 2011-05-11 | 南京中医药大学 | Novel heterocyclic compounds |
| CN102178697B (en) * | 2011-04-26 | 2013-01-02 | 李双阳 | Compound leech capsule and preparation method thereof |
| CN105606831A (en) * | 2016-01-29 | 2016-05-25 | 河南中医学院 | Method for detecting quality of thrombus clear freeze-dried powder needle based on blood agglutination activity |
| CN106729678A (en) * | 2016-12-29 | 2017-05-31 | 界首市菁华科技信息咨询服务有限公司 | A kind of complex preparation for treating thrombus |
| CN108498755B (en) | 2018-04-17 | 2021-04-09 | 贾洪章 | Chinese medicinal composition for treating cardiovascular and cerebrovascular diseases, and its preparation method and application |
| CN113073093A (en) * | 2021-04-25 | 2021-07-06 | 山东中医药大学 | Leech fibrinolytic active protein and preparation method thereof |
| CN113813291B (en) * | 2021-11-10 | 2023-05-26 | 鲁南制药集团股份有限公司 | Preparation method of animal medicinal material freeze-dried powder |
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| CN1238186A (en) * | 1998-06-10 | 1999-12-15 | 陈松林 | Medicine for curing hemiplegia and its preparing process |
| CN1255336A (en) * | 1998-11-27 | 2000-06-07 | 常德市第一中医院 | Medicine for treating apoplexy and thrombus |
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| CN1238186A (en) * | 1998-06-10 | 1999-12-15 | 陈松林 | Medicine for curing hemiplegia and its preparing process |
| CN1255336A (en) * | 1998-11-27 | 2000-06-07 | 常德市第一中医院 | Medicine for treating apoplexy and thrombus |
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