3, summary of the invention
In order to meet clinical needs, better treat cardiovascular and cerebrovascular disease, the invention provides a kind of new pharmaceutical composition and its production and use.
Pharmaceutical composition of the present invention comprises Folium Ginkgo extract, Tanshinone I I
ASodium sulfonate.Through a large amount of screening test of inventor, prove Folium Ginkgo extract and Tanshinone I I
AThe preferred weight proportioning of sodium sulfonate is 1: 0.08~8, it for example can be 1: 0.08,1: 0.1,1: 0.16,1: 0.2,1: 0.3,1: 0.4,1: 0.5,1: 0.6,1: 0.7,1: 0.8,1: 0.9,1: 1,1: 1.5,1: 1.6,1: 2,1: 2.4,1: 2.5,1: 3,1: 3.2,1: 4,1: 5,1: 6,1: 7,1: 8, further optimum ratio is 1: 0.2~4, further be preferably 1: 0.4~and 2.
In the aforementioned pharmaceutical compositions, the content of total flavonoids is not less than 24.0% in the Folium Ginkgo extract, the content of terpene lactone is not less than 6.0%, and the content of total ginkgolic acids is no more than 10/1000000ths; The content that is preferably total flavonoids is not less than 24.0%, the content of terpene lactone is not less than 6.0%, and the content of total ginkgolic acids is no more than 1,000,000/.
Pharmaceutical composition of the present invention, the weight proportion of drug component is groped to sum up to draw through the inventor in a large number, and each component all has better curative effect in above-mentioned weight proportion scope.Above-mentioned weight proportion is for especial patient, the ratio of can corresponding adjustment forming.
The present invention also provides the purposes of aforementioned pharmaceutical compositions in the medicine of preparation resisting cardiovascular disease.Pharmacological evaluation shows; but pharmaceutical composition significant prolongation rat experiment carotid artery thrombus formation time of the present invention; collagen protein-epinephrine inducing mouse cerebral thrombosis death had remarkable protective effect; can significantly improve the Mice Auricle microcirculation; significantly improve the anesthetized open-chest dog hemodynamics; cerebral ischemia is had remarkable protective effect, significantly reduce the abnormal change of myocardial ischemia scope and electrocardio due to the ligation rat coronary artery, biochemical indicator.
The present composition can be made clinically any or pharmaceutically acceptable dosage form, optimizing injection or oral formulations with acceptable accessories; Can parenteral or mode such as oral administration be applied to the patient who needs this treatment.
When being used for parenteral, can be made into injection.Injection means the intravital solution of confession injection, emulsion or the suspension that medicine is made and supplies to face with preceding preparation or be diluted to solution or the sterile preparation of the powder of suspension or concentrated solution that injection can be divided into injection, injectable sterile powder and concentrated solution for injection.Injection means that the confession that medicine is made is injected into sterile solution type injection, emulsion-type injection or the suspension type injection of using in the body, can be used for intramuscular injection, intravenous injection, intravenous drip etc.; Its specification has 1ml, 2ml, 5ml, 10ml, 20ml, 50ml, 100ml, 200ml, 250ml, 500ml etc., and wherein large volume (generally the being not less than 100ml) injection of using for intravenous drip also claims venous transfusion.Injectable sterile powder means that confession that medicine is made is faced with the suitable sterile solution of preceding usefulness and is mixed with settled solution or the evenly sterilized powder or the aseptic block of suspension, available suitable solvent for injection preparation back injection, also available venous transfusion preparation posterior vein instils; Sterilized powder makes with solvent crystallization, spray drying method or freeze-drying etc.Concentrated solution for injection means that confession that medicine is made faces the aseptic concentrated solution of using for intravenous drip with preceding dilution.
When making injection, can adopt the conventional method production in the existing pharmaceutical field, optional use solvent or non-aqueous solvent.The most frequently used aqueous solvent is a water for injection, also available 0.9% sodium chloride solution or other suitable aqueous solutions; Non-aqueous solvent commonly used is a vegetable oil, is mainly the injection soybean oil, and other also have the aqueous solution of ethanol, propylene glycol, Polyethylene Glycol etc.During the preparation injection, can add suitable additives according to the character of medicine, as osmotic pressure regulator, pH value regulator, solubilizing agent, filler, antioxidant, antibacterial, emulsifying agent, suspending agent etc.Osmotic pressure regulator commonly used comprises sodium chloride, glucose, potassium chloride, magnesium chloride, calcium chloride, sorbitol etc., preferred sodium chloride or glucose; PH value regulator commonly used comprises acetic acid-sodium acetate, lactic acid, citric acid-sodium citrate, sodium bicarbonate-sodium carbonate etc.; Solubilizing agent commonly used comprises polyoxyethylene sorbitan monoleate, propylene glycol, lecithin, polyoxyethylene castor oil etc.; Filler commonly used comprises lactose, mannitol, sorbitol, dextran etc.; Antioxidant commonly used has sodium sulfite, sodium sulfite, sodium pyrosulfite etc.; Antibacterial commonly used is phenol, cresol, chlorobutanol etc.Injection container commonly used has glass ampule, vial, plastic ampoule, plastic bottle etc.
Be used for when oral, can be made into conventional solid preparation, as tablet, capsule, pill, granule etc.; Also can be made into oral liquid, as oral solution, oral suspensions, syrup etc.Tablet means disk shape or the special-shaped flaky solid preparation that medicine and the auxiliary materials and mixing compacting that suits form, based on oral ordinary tablet, other has buccal tablet, Sublingual tablet, mouth paster, chewable tablet, dispersible tablet, fuse, effervescent tablet, slow releasing tablet, controlled release tablet and enteric coatel tablets etc.Capsule means medicine or is added with the adjuvant filling in Capsules or be sealed in solid preparation in the soft capsule material, according to its dissolving and release characteristics, can be divided into hard capsule (being commonly referred to as capsule), soft capsule (soft gelatin capsule), slow releasing capsule, controlled release capsule and enteric coated capsule etc.Pill means medicine and suitable adjuvant uniform mixing, and the spherical or near-spherical solid preparation so that proper method is made comprises drop pill, sugar pill, piller etc.Granule means that medicine and suitable adjuvant make the dried particles shape preparation with certain particle size, can be divided into soluble particles (being commonly referred to as granule), mix suspension grain, effervescent granule, enteric coated particles, slow-releasing granules and controlled release granule etc.Oral solution means that medicine dissolution makes for oral supernatant liquid preparation in suitable solvent.Oral suspensions means the slightly solubility solid drugs, is dispersed in the liquid medium, makes for oral suspension body preparation, also comprises dry suspension or dense suspension.Syrup means the dense aqueous sucrose solution that contains medicine.
When making oral formulations, can add suitable filler, binding agent, disintegrating agent, lubricant etc.Filler commonly used comprises starch, Icing Sugar, calcium phosphate, calcium sulfate two water things, dextrin, microcrystalline Cellulose, lactose, pregelatinized Starch, mannitol etc.; Typical binders comprises sodium carboxymethyl cellulose, PVP-K30, hydroxypropyl cellulose, starch slurry, methylcellulose, ethyl cellulose, hypromellose, gelling starch etc.; Disintegrating agent commonly used comprises dried starch, polyvinylpolypyrrolidone, cross-linking sodium carboxymethyl cellulose, carboxymethyl starch sodium, low-substituted hydroxypropyl cellulose etc.; Conventional lubricants comprises magnesium stearate, Pulvis Talci, sodium lauryl sulphate, micropowder silica gel etc.
In the pharmaceutical composition of the present invention, the preferred for preparation technology of Folium Ginkgo extract is as follows:
Selection process one: get Folium Ginkgo and pulverize, add 60% aqueous acetone solution, 57~59 ℃ of vigorous stirring 30min, filter, repeat above-mentioned steps, extract again one time, merging filtrate, filtrate decompression are concentrated into and contain about 30%~40% (content about 5% of acetone at this moment) of solids.Thin up is to 2 times of original volume, be cooled to about 12 ℃ under stirring, be incubated 1 hour, centrifugal, discard precipitation, supernatant adds ammonium sulfate by 30%, stir evenly, make molten entirely, with butanone-acetone mixed solution (6: 4) extraction 2 times, each is 0.5 times of amount of aqueous solution, butanone-acetone mixed solution is evaporated to and contains solids 50%~70%, adds entry and makes that to contain solids about 10%, with the water-saturated n-butanol extraction of 1/2 volume 3 times, n-butyl alcohol liquid is evaporated to and contains solids more than at least 50%, add suitable quantity of water then and continue again to concentrate, in the gained concentrated solution, add entry and ethanol, make solid contents reach 10%, determining alcohol reaches 30%, add n-hexane extraction 3 times, each 1/3 amount, water concentrates, and drying under reduced pressure (60~80 ℃), promptly.Content by total flavonoids in the Folium Ginkgo extract of above-mentioned prepared is not less than 24.0%, and the content of terpene lactone is not less than 6.0%.
Selection process two: get Folium Ginkgo, pulverize, earlier with 12 times of amount 60% alcoholic solution reflux secondaries, each 3 hours, filter, merge ethanol extract, be condensed into thick paste, become every 1ml to be equivalent to the solution of 0.5g crude drug with the hot water dissolving, leave standstill cooling, filter macroporous resin on the filtrate (DA201) post, use 18% respectively, 30%, and the ethanol elution of 50% concentration, 18%, 30% ethanol takes off liquid and is concentrated into nothing alcohol flavor, and last polyamide column is with 95% ethanol elution, 50% ethanol elution of macroporous resin and the eluent of polyamide column are merged, be concentrated into no ethanol flavor,, discard the cyclohexane extraction extract with the cyclohexane extraction extraction, the concentrated solution spray drying, promptly.Content by total flavonoids in the Folium Ginkgo extract of above-mentioned prepared is not less than 24.0%, and the content of terpene lactone is not less than 6.0%.
In the aforementioned pharmaceutical compositions, Folium Ginkgo extract can also be by above-mentioned prepared, but this should be interpreted as Folium Ginkgo extract only limits to above-mentioned preparation technology in the aforementioned pharmaceutical compositions.
In sum, pharmaceutical composition of the present invention has following advantage:
(1) provides a kind of new pharmaceutical composition that is used to prepare medicament against cardiovascular disease, satisfied clinical needs.
(2) consumption to pharmaceutical composition Chinese medicine component of the present invention has carried out groping in a large number research, by Folium Ginkgo extract and Tanshinone I I
AThe different proportionings of sodium sulfonate filter out the weight proportion scope with significant curative effect to the thrombotic influence of rat experiment carotid artery.
(3) pharmacological effect studies show that; but pharmaceutical composition significant prolongation rat experiment carotid artery thrombus formation time of the present invention; collagen protein-epinephrine inducing mouse cerebral thrombosis death had remarkable protective effect; can significantly improve the Mice Auricle microcirculation; significantly improve the anesthetized open-chest dog hemodynamics; cerebral ischemia is had remarkable protective effect, significantly reduce the abnormal change of myocardial ischemia scope and electrocardio due to the ligation rat coronary artery, biochemical indicator.Aspect resisting cardiovascular disease remarkable result being arranged, is that those of ordinary skills institute is beyond thought.
(4) in the pharmaceutical composition of the present invention, active constituent content is definite in the Folium Ginkgo extract, Tanshinone I I
AThe sodium sulfonate structure is clear and definite, determined curative effect, with Folium Ginkgo extract and Tanshinone I I
ASodium sulfonate directly feeds intake, and preparation technology is easy, has avoided because the shortcoming that the quality of the pharmaceutical preparations that the crude drug mass discrepancy causes differs greatly, and the quality of the pharmaceutical preparations improves a lot, and impurity content significantly reduces, and safety is higher.
(5) to the raw material or the preparation of pharmaceutical composition of the present invention, carried out discriminating, assay, stability study etc., effective ingredient is clear and definite, content is high, and better stability of preparation is convenient to control product quality, can guarantee clinical drug safety.
(6) provide preferred Folium Ginkgo extract preparation technology, simple and easy to do, and quality is better, is adapted to industrialized great production.
Below routine by experiment beneficial effect of further setting forth pharmaceutical composition of the present invention, these experimental examples comprise pharmacodynamic experiment, the stability experiment of new pharmaceutical compositions of the present invention.Pharmaceutical composition of the present invention has following beneficial effect, but this should be interpreted as that pharmaceutical composition of the present invention only has following beneficial effect.
Used Folium Ginkgo extract is commercial refining gained in the experimental example 1~4, and experimental example 5~7 used Folium Ginkgo extract are embodiment 1 preparation gained.Replace pharmaceutical composition of the present invention with YD in the following experimental example.
Experimental example 1 pharmaceutical composition of the present invention is to the thrombotic influence of rat experiment carotid artery
The animal subject male rat, 140, body weight 240~280g is divided into 14 groups at random, 10 every group.
Test sample Folium Ginkgo extract: contain Folium Ginkgo flavonol glycosides 24.6%, terpene lactone 6.2%;
Tanshinone I I
ASodium sulfonate: Arbeila Medicine Holding(Tonghua) Co., Ltd.;
Folium Ginkgo extract and Tanshinone I I
AEach proportioning injection of sodium sulfonate: self-control, preparation method is seen embodiment 2;
Injection Folium Ginkgo extract: 50mg contains Folium Ginkgo flavonoid glycoside 12mg, terpenoid 3mg, and German Weil-McLain relaxes
Training doctor pharmaceutical factory;
Tanshinone I I
ASodium sulfonate injection: 2ml: 10mg, Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd;
Sodium chloride injection: 100ml: 0.9g, Shandong Huaxin Pharmaceutical Co., Ltd..
Dosage sees Table 1.
The experimental technique rat is with 2.5% pentobarbital sodium (25mg/kg) intraperitoneal injection of anesthesia, dorsal position is fixed, separate right carotid, adopt electrical injuries carotid artery intima method, form instrument with the experimental thrombus in vivo of BT87-3 and measure different group animal carotid artery thrombus formation time.Stimulating electrode and temperature probe are hung on the common carotid artery, press intraperitoneal injection shown in the table 1,10min begins to stimulate after the administration, stimulus intensity is 2mA, closes the thorn energizing switch behind the stimulation 5min, takes off electrode, regulate temp controlled meter behind the 3min to zero-bit, observe the tremulous pulse temperature bust time.The record electricity irritation began to the time of aorta temperature bust, and this time is decided to be carotid artery thrombus formation time (surpassing 3000 seconds persons in 3000 seconds).
Table 1 pharmaceutical composition of the present invention is to the influence of rat experiment carotid artery thrombus formation time
(n=10, meansigma methods ± standard deviation)
Experimental result and conclusion experimental result see Table 1.The result shows, Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate weight proportion is 1: 0.08~1: 8 o'clock, all rat experiment thrombosis is had remarkable protective effect, and all is higher than injection Folium Ginkgo extract or Tanshinone I I
AThe effect of the independent medication of sodium sulfonate injection is compared the p value all less than 0.001 with the blank group, points out two medicines that synergistic function is arranged.According to the thrombus formation time of each group, Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate weight proportion is 1: 0.2~4 o'clock, better effects if; Weight proportion is 1: 0.4~2.4 o'clock, and effect is best.
The influence that experimental example 2 pharmaceutical compositions of the present invention form the mice thrombus in vivo
120 of animal subject male mices, body weight 20~25g is divided into 6 groups at random, 20 every group.
Test sample injection YD1:90mg contains Folium Ginkgo extract 50mg, Tanshinone I I
ASodium sulfonate 40mg;
Injection YD2:120mg contains Folium Ginkgo extract 100mg, Tanshinone I I
ASodium sulfonate 20mg;
Injection YD3:125mg contains Folium Ginkgo extract 25mg, Tanshinone I I
ASodium sulfonate 100mg;
Injection Folium Ginkgo extract: 50mg contains Folium Ginkgo flavonoid glycoside 12mg, terpenoid 3mg, and German Weil-McLain relaxes
Training doctor pharmaceutical factory;
Tanshinone I I
ASodium sulfonate injection: 2ml: 10mg, Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd;
Sodium chloride injection: 100ml: 0.9g, Shandong Huaxin Pharmaceutical Co., Ltd..
Dosage sees Table 2.
Behind the experimental technique gastric infusion 1 hour, tail vein injection collagen protein-every 0.1mg of epinephrine mixed thrombus derivant.Record mice hemiplegia is recovered number in the 20min of injection back, and calculates each administration group to the thrombotic protective rate of mouse brain [dead protective rate=(matched group death toll-administration group death toll)/matched group death toll].
The influence (n=20) that table 2 pharmaceutical composition of the present invention forms the mice thrombus in vivo
Experimental result and conclusion experimental result see Table 2.The result shows, injection YD1, injection YD2, injection YD3, injection Folium Ginkgo extract, Tanshinone I I
AThe sodium sulfonate injection all has protective effect to collagen protein-epinephrine inducing mouse cerebral thrombosis death.According to dead protective rate, Folium Ginkgo extract and Tanshinone I I
AThe proportioning of sodium sulfonate is 1: 0.8,1: 0.2,1: 4 o'clock, and the protective effect to collagen protein-epinephrine inducing mouse cerebral thrombosis death all is higher than injection Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate injection, drug effect is remarkable, points out two medicines that synergistic function is arranged.
Experimental example 3 pharmaceutical compositions of the present invention influence microcirculation of mouse auricle
The animal subject male mice, 60, body weight 25~28g is divided into 6 groups at random, 10 every group.
Test sample YD injection 1:5ml: 90mg contains Folium Ginkgo extract 50mg, Tanshinone I I
ASodium sulfonate 40mg;
YD injection 2:5ml: 115mg contains Folium Ginkgo extract 75mg, Tanshinone I I
ASodium sulfonate 40mg;
YD injection 3:5ml: 135mg contains Folium Ginkgo extract 75mg, Tanshinone I I
ASodium sulfonate 60mg;
Injection Folium Ginkgo extract: 50mg contains Folium Ginkgo flavonoid glycoside 12mg, terpenoid 3mg, and German Weil-McLain relaxes
Training doctor pharmaceutical factory;
Tanshinone I I
ASodium sulfonate injection: 2ml: 10mg, Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd;
Sodium chloride injection: 100ml: 0.9g, Shandong Huaxin Pharmaceutical Co., Ltd..
Dosage sees Table 3.
After the administration of experimental technique mouse stomach, with urethane 1g/kg intraperitoneal injection of anesthesia.Postanesthetic mice places on the supporting plate that is covered with Cotton Gossypii, keeps 20 ± 2 ℃ of room temperatures.Mouse right ear is placed on the ear carriage, on auris dextra, drips a little liquid paraffin, supporting plate is placed on the microscope carrier, regulate cold light source, make illuminating ray and auricle plane at 45~60 ° of angles, and parallel with the direction of growth of hair.Under low power lens, observe the auricle overall picture earlier, select suitable position, use high power lens instead and fix a point to observe continuously.The microcirculating state of 20min after the observation administration, the diameter and the blood flow rate of measurement arteriole, venule.
Table 3 Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate drug combination influences microcirculation of mouse auricle
(meansigma methods ± standard deviation, n=10)
*P<0.05;
*Compare with matched group in p<0.01.
Experimental result and conclusion experimental result see Table 3.The result shows, YD injection, injection Folium Ginkgo extract, Tanshinone I I
AThe sodium sulfonate injection all makes mouse ear arteriole, venule caliber increase, and blood flow rate is accelerated.Especially YD injection administration group, ear arteriole, venule caliber obviously increase (p<0.01), and blood flow rate is significantly accelerated (p<0.01).And the YD injection all is better than injection Folium Ginkgo extract and Tanshinone I I to the microcirculation of mouse auricle effect
AThe effect of sodium sulfonate injection points out two medicines that synergistic function is arranged.
Experimental example 4 pharmaceutical compositions of the present invention are to the hemodynamic influence of anesthetized open-chest dog
Animal subject hybrid dog, 25, body weight 11.0~13.0kg is divided into 5 groups at random, 5 every group.
Test sample YD injection 1:5ml: 90mg contains Folium Ginkgo extract 50mg, Tanshinone I I
ASodium sulfonate 40mg;
Injection YD1:90mg contains Folium Ginkgo extract 50mg, Tanshinone I I
ASodium sulfonate 40mg;
Injection Folium Ginkgo extract: 50mg contains Folium Ginkgo flavonoid glycoside 12mg, terpenoid 3mg, and German Weil-McLain relaxes
Training doctor pharmaceutical factory;
Tanshinone I I
ASodium sulfonate injection: 2ml: 10mg, Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd
Sodium chloride injection: 100ml: 0.9g, Shandong Huaxin Pharmaceutical Co., Ltd..
Dosage 5mg/kg
After the experimental technique dog is anaesthetized with pentobarbital sodium (30mg/kg) intravenous injection, take under the right arm reclining malleation artificial respiration, between 4~5 sides of body of a left side, open breast, open pericardium in distance vagus nerve 2cm place, parietal layer is made outstanding bed it is sutured in thoracic wall, and heart is fully exposed.Separate aorta, aortic root be inserted in the electromagnetic flowmeter probe (10~12mm), measure cardiac output (CO); At the left anterior descending coronary artery root, separate visceral pericardium, isolate about 1cm coronary artery, be inserted in the electromagnetic flowmeter probe (2~3mm), measurement coronary flow (CBF); This two probe is connected on the LMTC-621 type electromagnetic flowmeter, separates a bilateral common carotid artery, intubate connects pressure transducer, record arteriotony (AP) and mean arterial pressure (MAP); With internal diameter is that the cardiac catheter of 1.5mm is inserted left ventricle from the apex of the heart, amplify left indoor pressure (LVP) by YZ-1 type pressure transducer through carrier wave, the LVP electric signal amplifies 10 times through direct current amplifier, write down left chamber EDP (LVEDP), with the LVP electric signal again through BMI type differentiator derivative recording left indoor pressure rate of change (dp/dt
Max), it is subcutaneous to insert the animal subject extremity with pin type electrode, and record mark II lead electrocardiogram (EGG-II) is to measure heart rate (HR).These parameters changes equal synchronous recording and leads instrument in RM-6300 physiology more.
Separate femoral artery and take out arterial blood, through External Carotid Artery for Intubation to the coronary sinus vein venous blood samples, according to CY-2 oxygen analyser operation instructions, with freshly prepared anaerobic solution (crystallization of 0.01M borax soln adding anhydrous sodium sulfite is mixed with 2% sodium sulfite solution) zeroing, use with the distilled water of air balance temperature constant and make sensitivity adjusting.Treat to measure oxygen content with oxygen analyser behind the instrument stabilizer.Experiment finishes back calculating myocardium oxygen consumption.
After operation finishes, observe above-mentioned every index, after stable with the index of record value before as administration, behind the intravenously administrable in 1,3,5,10,20,30min, gather above-mentioned every index, and with every index rate of change (%) of each time point after the administration, the t-test that does significance pairing data between group with every index rate of change (%) of each corresponding time point of matched group handles.
Experimental result and conclusion YD injection 1, injection YD1, injection Folium Ginkgo extract, Tanshinone I I
AThe sodium sulfonate injection is to being tried dog heart rate (HR), mean arterial pressure (MBP), left indoor pressure (LVP) and ventricular muscles contractility (dp/dt
Max), myocardial oxygen consumption all has the reduction effect.Especially YD injection 1 and injection YD1 organize, and the reduction amplitude is all obviously greater than injection Folium Ginkgo extract group and Tanshinone I I
ASodium sulfonate injection group, myocardial oxygen consumption reduce in beginning about 2 minutes after the administration, and maximum is respectively 25.1% and 25.3%, reach action time more than 35 minutes, with matched group the curative effect (p<0.01) of highly significant are arranged relatively.Prompting, Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate drug combination has synergistic function.
Experimental example 5 pharmaceutical compositions of the present invention are to the protective effect of cerebral ischemia
The animal subject rat, 140, the male and female dual-purpose, body weight 200~220g is divided at random: sham operated rats, ischemia-reperfusion group, 4 groups of YD injection, 5 groups of YD injection, 6 groups of YD injection, gingko leaf extract injection, Tanshinone I I
ASodium sulfonate injection group, 20 every group.
Test sample YD injection 4:5ml: 75mg contains Folium Ginkgo extract 25mg, Tanshinone I I
ASodium sulfonate 50mg;
YD injection 5:5ml: 70mg contains Folium Ginkgo extract 50mg, Tanshinone I I
ASodium sulfonate 20mg;
YD injection 6:5ml: 72mg contains Folium Ginkgo extract 40mg, Tanshinone I I
ASodium sulfonate 32mg;
Gingko leaf extract injection: self-control, 2ml: 50mg, preparation method is referring to embodiment 2;
Tanshinone I I
AThe sodium sulfonate injection: self-control, 2ml: 10mg, preparation method is referring to embodiment 2;
Sodium chloride injection: 100ml: 0.9g, Shandong Huaxin Pharmaceutical Co., Ltd..
Dosage sees Table 4.
The experimental technique rat is with 10% chloral hydrate 3ml/kg intraperitoneal injection of anesthesia, neck median incision, separation, ligation right carotid proximal part, external carotid artery and bifurcated artery thereof.Separate the right side internal carotid artery, separate wing jaw tremulous pulse downwards along internal carotid artery, this branch of root ligation.Be equipped with line, far-end placement bulldog clamp at the internal carotid artery near-end, common carotid artery crotch otch, (degree of depth is 17~20mm), fastens line and enters internal carotid artery, goes into cranium to anterior cerebral artery, all blood flows sources of blocking-up middle cerebral artery to insert nylon wire.Remove bulldog clamp, tighten line fully, stay the long the end of a thread of 1cm outward, skin suture.Gastric infusion or sodium chloride injection behind the ischemia 1h continue perfusion again behind the ischemia 1h, need not anaesthetize and cut skin once more, and prompting nylon wire head end is to the common carotid artery incision when resistance is arranged to lift institute's the end of a thread that stays gently, and blood flow is logical again.Sham operated rats is except that plug wire not, and all the other steps are the same.After the survival Mus is poured into 24h again, observe rat behavior and change, carry out behavior scoring.5 fens system standards of grading with reference to zea Longa: 0 minute, normal, impassivity damage symptom; 1 minute, can not full extension offside fore paw; 2 minutes, turn-take laterally; 3 minutes, topple over to offside; 4 minutes, can not spontaneously walk loss of consciousness.Broken end is got the Mus brain fast then.A part (10 every group) is divided another name left and right sides brain hemisphere weight in wet base, puts in 160 ℃ of baking boxs to claim dry weight behind the 24h, calculates brain water content as follows: brain water content (%)=(weight in wet base-dry weight)/weight in wet base * 100%; A part (10 every group) is downcut the crown brain sheet of thick about 2mm on the anterior commissure plane, place 2%TIE solution at once, hatches 30min for 37 ℃.Infarct presents white, and non-infarct presents redness.Measure with planimeter (C63 image analysis system) and respectively to distinguish area, and calculate the percentage ratio (%) that infarct accounts for full brain.
Experimental result and conclusion experimental result see Table 4.
(1) to the influence of behavior: all no abnormal symptom of sham operated rats rat, the neuroethology scoring is 0.The sodium chloride injection matched group occurs can not full extension offside fore paw or the nerve injury symptom of turn-taking laterally or toppling over to offside, and behavior scoring is 1.7 ± 0.8.Compare with the sodium chloride injection matched group, YD injection 4, YD injection 5, YD injection all can extremely significantly reduce the neuroethology scoring for 6 groups, are respectively 0.7 ± 0.5,0.6 ± 0.4,0.5 ± 0.4 (p<0.01); The gingko leaf extract injection group can significantly reduce the neuroethology scoring, is 0.8 ± 0.5 (p<0.05); Tanshinone I I
ASodium sulfonate injection group can reduce the neuroethology scoring, is 0.9 ± 0.6 (p<0.05).
(2) to the influence of brain water content: the brain water content utmost point of sodium chloride injection matched group ischemia-reperfusion side (right hemisphere) is significantly higher than sham operated rats (p<0.01), and the modeling success is described.Compare with the sodium chloride injection matched group, the brain water content that YD injection 4, YD injection 5, YD injection are 6 groups all extremely significantly reduces (p<0.01), gingko leaf extract injection group and Tanshinone I I
AThe brain water content of sodium sulfonate injection group all significantly reduces (p<0.05).
(3) to the influence of infarct size: the sham operated rats cerebral tissue does not have infraction.Sodium chloride injection matched group ischemia side cerebral tissue has the infraction phenomenon, and infarct volume accounts for 37.33% of full brain.Compare with the sodium chloride injection matched group, YD injection 4, YD injection 5, YD injection 6 all can extremely significantly dwindle ischemia side cerebral tissue infarct volume (p<0.01), gingko leaf extract injection and Tanshinone I I
AThe sodium sulfonate injection all can significantly dwindle ischemia side cerebral tissue infarct volume (p<0.05).
Table 4 pharmaceutical composition of the present invention is to the protective effect of cerebral ischemia
(meansigma methods ± standard deviation, n=10)
*P<0.05,
*Compare with the sodium chloride injection matched group in p<0.01;
*Compare with sham operated rats in p<0.01.
Above-mentioned experimental result shows, Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate drug combination can significantly improve the nerve injury symptom of focal brain ischemia-reperfusion injury, reduces the ischemical reperfusion injury brain water content, alleviates ischemia side brain hemisphere edema degree and dwindles cerebral infarct volume.Folium Ginkgo extract and Tanshinone I I
ASodium sulfonate is 1: 2,1: 0.4,1: 0.8 o'clock, and the effect of each treatment group all is better than gingko leaf extract injection or Tanshinone I I
AThe effect of the independent medication of sodium sulfonate injection, prompting Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate drug combination has synergistic function.
Experimental example 6 pharmaceutical compositions of the present invention are to the influence of ligation rat platelet agglutination
The animal subject rat, 140, body weight 200~220g, the male and female dual-purpose is divided into 14 groups at random, 10 every group.
The test sample Folium Ginkgo extract: self-control, see embodiment 1;
Tanshinone I I
ASodium sulfonate: Arbeila Medicine Holding(Tonghua) Co., Ltd.;
Folium Ginkgo extract and Tanshinone I I
AThe solution of the different proportionings of sodium sulfonate: self-control (proportioning sees Table 5);
Normal saline: self-control.
Dosage: see Table 5.
Each treated animal of experimental technique is pressed table 5 gastric infusion, every day 1 time, and successive administration 7 days, the last administration is after 1 hour, with 30% pentobarbital sodium 30mg/kg intraperitoneal injection of anesthesia, from abdominal aortic blood, with 3.28% sodium citrate anticoagulant (mixing with 1: 9 with whole blood).The centrifugal 5min of anticoagulated whole blood 1500rpm under 20 ℃ of conditions is obtained platelet rich plasma (PPR).After leaving and taking quantitative PPR, will remain PPR, obtain own control platelet poor plasma (PPP) once more with the centrifugal 10min of 3000rpm.Regulate PPR concentration with PPP, make each PPR concentration identical.In 37 ℃ constant temperature hole after the preheating, (ADP, final concentration are 3 μ molL to add adenosine diphosphate (ADP) with PPR
-1) the induced platelet gathering, the record maximum agglutination rate, and calculate platelet aggregation suppression ratio=(matched group coagulation rate-medicine group coagulation rate)/matched group coagulation rate * 100%.
Experimental result and conclusion experimental result see Table 5.Compare Folium Ginkgo extract, Tanshinone I I with the sodium chloride injection matched group
ASodium sulfonate all significantly suppresses platelet aggregation (p<0.05); Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate weight proportion is 1: 0.08,1: 0.1,1: 6,1: 8 o'clock, all significantly suppresses platelet aggregation (p<0.05); Folium Ginkgo extract and Tanshinone I I
AThe sodium sulfonate weight proportion is 1: 0.2,1: 0.4,1: 0.6,1: 0.8,1: 1,1: 2,1: 4 o'clock, all extremely significantly suppresses platelet aggregation (p<0.01), and is especially best with 1: 0.4,1: 0.6,1: 0.8,1: 1,1: 2 o'clock effect.Folium Ginkgo extract and Tanshinone I I
AThe effect of each proportioning of sodium sulfonate all is better than Folium Ginkgo extract or Tanshinone I I
AThe effect that sodium sulfonate is individually dosed, prompting Folium Ginkgo extract and Tanshinone I I
ASodium sulfonate has synergistic function.
Table 5 pharmaceutical composition of the present invention is to the influence of rat platelet agglutination
*P<0.01,
*Compare with the sodium chloride injection matched group in p<0.001.
Experimental example 7 pharmaceutical compositions of the present invention are to the influence of myocardial ischemia due to the ligation rat coronary artery
The animal subject rat, 60, body weight 200~220g, the male and female dual-purpose is divided into 6 groups at random, 10 every group.
Test sample YD granule: 80mg contains Folium Ginkgo extract 40mg, Tanshinone I I
ASodium sulfonate 40mg (embodiment 11);
Folium Ginkgo extract: self-control, see embodiment 1;
Tanshinone I I
ASodium sulfonate: Arbeila Medicine Holding(Tonghua) Co., Ltd.;
Normal saline: self-control.
Dosage sees Table 6.
The experimental technique rat is used urethane 1g/kg intraperitoneal injection of anesthesia, back of the body position is fixing, the record electrocardio connects artificial respirator and practices artificial respiration, and opens the thoracic cavity, cut off pericardium, each treated animal is pressed table 6 gastric infusion, falls branch before the coronary artery of ligation left side behind the 3min, omnidistance record electrocardio 30min, 1h gets blood after the ligation, detects creatine phosphokinase (CK) and lactic acid dehydrogenase (LDH).Take out rat heart, with 4 of the even crosscuts of ventricular muscles, 0.5% chlorination nitro tetrazole is blue to dye along ligature, and with the ischemic areas on every myocardium two sides of planimeter survey, the calculating myocardium ischemic areas accounts for the percentage ratio of ventricle area.
Table 6 pharmaceutical composition of the present invention is to the influence of myocardial ischemia due to the ligation rat coronary artery
(meansigma methods ± standard deviation, n=10)
*P<0.05,
*Compare with the myocardial ischemia matched group in p<0.01.
Experimental result and conclusion experimental result see Table 6.After the ligation, the cardiac electrical QRS ripple of myocardial ischemia control rats all increases unusually suddenly, widens, and cardiac muscle is ischemia on a large scale, and biochemistry detection shows as CK and LDH all increases unusually, and the modeling success is described.Compare with the myocardial ischemia matched group, the YD granule of each dosage group all can extremely significantly reduce extremely significantly to reduce myocardial ischemia scope (p<0.01) because of the electrocardio due to the ligation coronary artery and the abnormal change (p<0.01) of biochemical indicator; Folium Ginkgo extract can extremely significantly reduce significantly to reduce myocardial ischemia scope (p<0.05) because of the electrocardio due to the ligation coronary artery and the abnormal change (p<0.01) of biochemical indicator; Tanshinone I I
ASodium sulfonate can extremely significantly reduce significantly to reduce myocardial ischemia scope (p<0.05) because of the electrocardio due to the ligation coronary artery and the abnormal change (p<0.01) of biochemical indicator.The particulate effect of the YD of each dosage group all is better than Folium Ginkgo extract or Tanshinone I I
AThe effect that sodium sulfonate is individually dosed, prompting Folium Ginkgo extract and Tanshinone I I
ASodium sulfonate has synergistic function.
Experimental example 8 pharmaceutical composition stability experiments of the present invention
Test sample YD injection: 5ml: 90mg contains Folium Ginkgo extract 50mg, Tanshinone I I
ASodium sulfonate 40mg
Investigation project: character, pH value, clarity, related substance, sign content; And at accelerated tests 6 months and the aseptic and pyrogen test of long-term experiment end of term increase.
1, influence factor's experiment
Test sample is got in the strong illumination experiment, and putting illumination is interior the placement 10 days of lighting box of 4500Lx.
Test sample is got in the high temperature experiment, places respectively under 40 ℃, the 60 ℃ conditions and places 10 days.
Low temperature test is got test sample, places 10 days in 4 ℃ of refrigerators.
Above-mentioned experiment was respectively at the 5th, 10 day sampling and measuring.Relatively test every index after the character, and with result and comparison in 0 day.
Placed 10 days under experimental result illumination 4500Lx, 60 ℃ of conditions of high temperature, remove related substance and slightly raise, outside sign content slightly descended, all other indexs had no significant change.Placed 10 days under 40 ℃ of high temperature, 4 ℃ of conditions of low temperature, every index does not have significant change.
2, accelerated tests
Experimental technique is put under the condition of 40 ℃ ± 2 ℃ of temperature, relative humidity 75% ± 5% and was placed 6 months.Respectively at taking a sample 1st month, 2 months, 3 months, 6 the end of month, relatively after the outward appearance, test every index at experimental session, with result and comparison in 0 month; And at 6 aseptic and pyrogen tests of increase at the end of month.
Placed under the condition of 40 ℃ ± 2 ℃ of experimental result temperature, relative humidity 75% ± 5% 6 months, except that related substance slightly increased, all other indexs had no significant change, and at 6 the end of month of accelerated tests, pyrogen, sterility test are all up to specification.
3, long-term experiment
Experimental technique is put under the condition of 25 ℃ ± 2 ℃ of temperature, relative humidity 60% ± 10% and was placed 18 months.Respectively at 3rd month, 6 months, 9 months, 12 months, 18 months, relatively after the outward appearance, test every index, with result and comparison in 0 month; And at 18 aseptic and pyrogen tests of increase at the end of month.
Placed under the condition of 25 ℃ ± 2 ℃ of experimental result temperature, relative humidity 60% ± 10% 18 months, every index has no significant change, and at 18 the end of month of long-term experiment, pyrogen, sterility test are all up to specification.
Conclusion is reached a conclusion by above-mentioned investigation result, and in every experiment, the YD injection is all more stable.
4, the specific embodiment
The specific embodiment of form is described in further detail foregoing of the present invention by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following embodiment.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.Adjuvant in following examples in the preparation of each dosage form can be replaced with acceptable accessories, perhaps reduces, increases.
The preparation of embodiment 1 Folium Ginkgo extract and discriminating and assay
The preparation of Folium Ginkgo extract
Get Folium Ginkgo, pulverize, earlier with 12 times of amount 60% alcoholic solution reflux secondaries, each 3 hours, filter, merge ethanol extract, be condensed into thick paste, become every 1ml to be equivalent to the solution of 0.5g crude drug with the hot water dissolving, leave standstill cooling, filter macroporous resin on the filtrate (DA201) post, use 18% respectively, 30%, and the ethanol elution of 50% concentration, 18%, 30% ethanol takes off liquid and is concentrated into nothing alcohol flavor, and last polyamide column is with 95% ethanol elution, 50% ethanol elution of macroporous resin and the eluent of polyamide column are merged, be concentrated into no ethanol flavor,, discard the cyclohexane extraction extract with the cyclohexane extraction extraction, the concentrated solution spray drying, promptly.
Prepare three batches of Folium Ginkgo extract according to the method described above respectively, yield sees Table 7.
The discriminating of Folium Ginkgo extract
(1) get Folium Ginkgo extract 0.2g, add n-butyl alcohol 15ml, put in the water-bath warm macerating 15 minutes and jolting constantly, put coldly, filter the filtrate evaporate to dryness, residue adds ethanol 2ml makes dissolving, as need testing solution.Other gets Folium Ginkgo reference extract 0.2g, shines extract solution in pairs with legal system.Test according to thin layer chromatography, draw each 3 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution that contains 0.4% sodium acetate, with ethyl acetate-butanone-methanol-water (5: 3: 1: 1) be developing solvent, launch, take out, dry, spray is put under the ultra-violet lamp (365nm) and is inspected with 3% aluminum chloride alcoholic solution.In the test sample chromatograph, with the corresponding position of reference extract chromatograph on, show the fluorescence speckle of same color.
(2) according to the thin layer chromatography test, terpene lactone need testing solution under the assay item of absorption terpene lactone and each 15 μ l of reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution that contains 0.4% sodium acetate, with toluene-ethyl acetate-acetone-methanol (10: 5: 5: 0.6) be developing solvent, launching below 15 ℃, take out, dry, smoked 15 minutes with the acetic anhydride steam, 140~160 ℃ of heating 30 minutes, put coldly, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
Three batches of Folium Ginkgo extract to above-mentioned preparation carry out identification experiment, all meet the requirements.
The assay of Folium Ginkgo extract
The assay of total pyrite alcohol glycosides
High performance liquid chromatography
Chromatographic condition and system suitability experiment are filler with the octadecylsilane chemically bonded silica; With methanol-0.4% phosphoric acid solution (50: 50) is mobile phase; The detection wavelength is 360nm.Number of theoretical plate calculates by the Quercetin peak should be not less than 2500.
The preparation of reference substance solution precision respectively takes by weighing Quercetin reference substance, kaempferide reference substance, isorhamnetin reference substance, adds methanol and makes the mixed solution that every 1ml contains 30 μ g, 30 μ g, 20 μ g respectively, product solution in contrast.
The about 35mg of Folium Ginkgo extract is got in the preparation of need testing solution, the accurate title, decide, the mixed solution 25ml that adds methanol-25% hydrochloric acid solution (4: 1) put in the water-bath reflux 30 minutes, was cooled to room temperature rapidly, be transferred in the 50ml measuring bottle, be diluted to scale with methanol, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and calculate the content of Quercetin, kaempferide and isorhamnetin respectively, are converted into the content of total pyrite alcohol glycosides by following formula.
Total pyrite alcohol glycosides=(quercetin content+kaempferide content+isorhamnetin content) * 2.51
The assay of terpene lactone
High performance liquid chromatography
Chromatographic condition and system suitability experiment are filler with the octadecylsilane chemically bonded silica; With normal propyl alcohol-oxolane-water (1: 15: 84) is mobile phase; Detect with evaporative light scattering detector.Number of theoretical plate calculates by the bilobalide peak should be not less than 2500.
The preparation of reference substance solution respectively precision to take by weighing bilobalide reference substance, ginkalide A reference substance, ginkalide B reference substance and ginkalide C reference substance an amount of, add methanol and make the mixed solution that every 1ml contains 2mg, 1mg, 1mg, 1mg, brilliant in contrast solution.
The about 0.15g of Folium Ginkgo extract is got in the preparation of need testing solution, and accurate the title decides, and adds water 10ml, puts to heat in the water-bath to make molten loosing, add 2 of 2% hydrochloric acid solutions, extract 4 (15ml, 10ml, 10ml with the ethyl acetate jolting, 10ml), merge extractive liquid, washs with 5% sodium acetate solution 20ml.Combined ethyl acetate extracting solution and washing liquid wash with water 2 times, and each 20ml divides water intaking liquid, with ethyl acetate 10ml washing, combined ethyl acetate liquid, reclaim solvent to doing, residue is with dissolve with methanol and be transferred in the 5ml measuring bottle, add methanol to scale, shake up, get subsequent filtrate, promptly.
Respectively accurate reference substance solution and need testing solution 5 μ l, the 10 μ l of drawing of algoscopy inject chromatograph of liquid, and mensuration is calculated the content of bilobalide, ginkalide A, ginkalide B and ginkalide C respectively with external standard two-point method logarithmic equation, promptly.
Three batches of Folium Ginkgo extract to above-mentioned preparation carry out assay respectively, and measurement result sees Table 7.
The assay result and the yield of table 7 Folium Ginkgo extract
The preparation of embodiment 2 medicine composition injections of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 40g
Polyoxyethylene sorbitan monoleate 50g
Water for injection adds to 5000ml
Prepare 1000 altogether
Prescription 2
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 40g
Polyoxyethylene sorbitan monoleate 75g
Water for injection adds to 5000ml
Prepare 1000 altogether
Prescription 3
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 60g
Polyoxyethylene sorbitan monoleate 75g
Water for injection adds to 5000ml
Prepare 1000 altogether
Prescription 4
Folium Ginkgo extract 25g
Tanshinone I I
ASodium sulfonate 50g
Polyoxyethylene sorbitan monoleate 60g
Water for injection adds to 5000ml
Prepare 1000 altogether
Prescription 5
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 20g
Polyoxyethylene sorbitan monoleate 60g
Water for injection adds to 5000ml
Prepare 1000 altogether
Prescription 6
Folium Ginkgo extract 40g
Tanshinone I I
ASodium sulfonate 32g
Polyoxyethylene sorbitan monoleate 60g
Water for injection adds to 5000ml
Prepare 1000 altogether
2, concrete steps:
(1) carries and handle the previous day such as pipeline that dosing uses and container etc., face with the fresh water for injection flushing of preceding reuse;
(2) polyoxyethylene sorbitan monoleate is made 20% aqueous solution, added the Folium Ginkgo extract of recipe quantity, the heated and stirred dissolving fully; Tanshinone I I
AIt is complete that sodium sulfonate adds the water for injection heating for dissolving of dosing amount 40%; Benefit adds to the full amount of water for injection;
(3) needle-use activated carbon of adding dosing amount 0.1%, heated and stirred 15 minutes;
(4) through sand filtration rod filtering decarbonization, measure the also pH value of regulator solution;
(5) through the microporous filter membrane fine straining of 0.45 μ m;
(6) clarity of inspection solution, the semi-finished product chemical examination;
(7) with the solution sealing by fusing in glass ampule;
(8) 100 ℃ of flowing steam sterilizations 30 minutes;
(9) while hot sample being put into 0.01% methylene blue solution hunts leak;
(10) lamp inspection;
(11) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 3 pharmaceutical composition powder pins of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 40g
Polyoxyethylene sorbitan monoleate 50g
Mannitol 700g
Sterile water for injection adds to 3000ml
Prepare 1000 altogether
Prescription 2
Folium Ginkgo extract 100g
Tanshinone I I
ASodium sulfonate 20g
Polyoxyethylene sorbitan monoleate 25g
Mannitol 400g
Sterile water for injection adds to 4000ml
Prepare 1000 altogether
Prescription 3
Folium Ginkgo extract 25g
Tanshinone I I
ASodium sulfonate 100g
Polyoxyethylene sorbitan monoleate 75g
Mannitol 700g
Sterile water for injection adds to 4000ml
Prepare 1000 altogether
Prescription 4
Folium Ginkgo extract 40g
Tanshinone I I
ASodium sulfonate 40g
Polyoxyethylene sorbitan monoleate 75g
Mannitol 700g
Sterile water for injection adds to 4000ml
Prepare 1000 altogether
2, concrete steps:
(1) vessel of at first dosing being used and antibiotic glass bottle, plugs etc. carry out aseptic process;
(2) take by weighing supplementary material according to recipe quantity;
(3) polyoxyethylene sorbitan monoleate is made 20% aqueous solution, added the Folium Ginkgo extract of recipe quantity, the heated and stirred dissolving fully; Tanshinone I I
AIt is complete that sodium sulfonate adds the water for injection heating for dissolving of dosing amount 40%; Add the dissolving of mannitol heated and stirred more fully, add sterile water for injection to full dose;
(4) needle-use activated carbon of adding dosing amount 0.1%, heated and stirred 15 minutes;
(5) through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution;
(6) through the microporous filter membrane fine straining of 0.22 μ m;
(7) clarity of inspection solution, the semi-finished product chemical examination;
(8) be sub-packed in the antibiotic glass bottle half tamponade; Sample is put into the freeze dryer lyophilization;-45 ℃ of pre-freezes 5 hours, low-temperature vacuum drying-45 ℃~0 ℃ 20 hours was warming up to 25 ℃ of vacuum dryings 3 hours then;
(9) lyophilizing finishes, and lid is rolled in tamponade;
(10) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 4 pharmaceutical composition sodium chloride injections of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 60g
Polyoxyethylene sorbitan monoleate 75g
Sodium chloride 900g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
Prescription 2
Folium Ginkgo extract 100g
Tanshinone I I
ASodium sulfonate 60g
Polyoxyethylene sorbitan monoleate 80g
Sodium chloride 900g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
Prescription 3
Folium Ginkgo extract 125g
Tanshinone I I
ASodium sulfonate 40g
Polyoxyethylene sorbitan monoleate 125g
Sodium chloride 900g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
Prescription 4
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 60g
Polyoxyethylene sorbitan monoleate 100g
Sodium chloride 900g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
2, concrete steps:
(1) handles the previous day such as pipeline that dosing uses and container etc., face with the fresh water for injection flushing of preceding reuse;
(2) polyoxyethylene sorbitan monoleate is mixed with 20% aqueous solution, adds the Folium Ginkgo extract of recipe quantity, the heated and stirred dissolving fully; Tanshinone I I
AIt is complete that sodium sulfonate adds the water for injection heating for dissolving of dosing amount 40%; Sodium chloride is complete with the water for injection dissolving of dosing amount 40%; Merge each solution, benefit adds to the full amount of water for injection;
(3) needle-use activated carbon of adding dosing amount 0.1%, heated and stirred 15 minutes;
(4) through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution;
(5) through the microporous filter membrane fine straining of 0.45 μ m;
(6) clarity of inspection solution, the semi-finished product chemical examination;
(7) fill is in the infusion bottle of 100ml;
(8) 115 ℃ of pressure sterilizings 30 minutes;
(9) lamp inspection;
(10) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 5 pharmaceutical composition glucose injections of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 100g
Tanshinone I I
ASodium sulfonate 80g
Polyoxyethylene sorbitan monoleate 100g
Glucose 5000g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
Prescription 2
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 80g
Glucose 5000g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
Prescription 3
Folium Ginkgo extract 125g
Tanshinone I I
ASodium sulfonate 60g
Polyoxyethylene sorbitan monoleate 125g
Glucose 5000g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
Prescription 4
Folium Ginkgo extract 100g
Tanshinone I I
ASodium sulfonate 40g
Polyoxyethylene sorbitan monoleate 100g
Glucose 5000g
Water for injection adds to 100000ml
Prepare 1000 bottles altogether
2, concrete steps:
(1) carries and handle the previous day such as pipeline that dosing uses and container etc., face with the fresh water for injection flushing of preceding reuse;
(2) polyoxyethylene sorbitan monoleate is mixed with 20% aqueous solution, adds the Folium Ginkgo extract of recipe quantity, the heated and stirred dissolving fully; Tanshinone I I
AIt is complete that sodium sulfonate adds the water for injection heating for dissolving of dosing amount 40%; Glucose is complete with the water for injection dissolving of dosing amount 40%; Merge each solution, benefit adds to the full amount of water for injection;
(3) needle-use activated carbon of adding dosing amount 0.1%, heated and stirred 15 minutes;
(4) through sand filtration rod filtering decarbonization.Measure the also pH value of regulator solution;
(5) through the microporous filter membrane fine straining of 0.45 μ m;
(6) clarity of inspection solution, the semi-finished product chemical examination;
(7) fill is in the infusion bottle of 100ml;
(8) 115 ℃ of pressure sterilizings 30 minutes;
(9) lamp inspection;
(10) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 6 pharmaceutical composition sheets of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 40g
Starch 120.0g
Microcrystalline Cellulose 40.0g
The 2%HPMC aqueous solution is an amount of
Magnesium stearate 2.0g
Carboxymethylstach sodium 4.0g
Prepare 1000 altogether
Prescription 2
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 40g
Starch 120.0g
Microcrystalline Cellulose 40.0g
The 2%HPMC aqueous solution is an amount of
Magnesium stearate 2.0g
Carboxymethylstach sodium 4.0g
Prepare 1000 altogether
Prescription 3
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 20g
Starch 120.0g
Microcrystalline Cellulose 40.0g
The 2%HPMC aqueous solution is an amount of
Magnesium stearate 2.0g
Carboxymethylstach sodium 4.0g
Prepare 1000 altogether
Prescription 4
Folium Ginkgo extract 25g
Tanshinone I I
ASodium sulfonate 50g
Starch 110.0g
Microcrystalline Cellulose 40.0g
The 2%HPMC aqueous solution is an amount of
Magnesium stearate 2.0g
Carboxymethylstach sodium 4.0g
Prepare 1000 altogether
2, concrete steps:
(1) with Folium Ginkgo extract and Tanshinone I I
AIt is standby that sodium sulfonate was pulverized 100 mesh sieves;
(2) take by weighing supplementary material according to recipe quantity;
(3) hypromellose 2% the aqueous solution made soluble in water is standby;
(4) with Folium Ginkgo extract, Tanshinone I I
ASodium sulfonate, starch, microcrystalline Cellulose mix homogeneously, adding 2%HPMC aqueous solution is an amount of, stirs, and makes suitable soft material;
(5) cross 20 mesh sieve system granules;
(6) granule is dried under 60 ℃ condition;
(7) dry good granule adds magnesium stearate and carboxymethylstach sodium, crosses 18 mesh sieve granulate, mix homogeneously;
(8) sampling, the semi-finished product chemical examination;
(9) the sheet weight sheet of determining according to chemical examination;
(10) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 7 medicament composition capsules of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 40g
Starch 60.0g
Microcrystalline Cellulose 20.0g
The 2%HPMC aqueous solution is an amount of
Magnesium stearate 1.0g
Prepare 1000 altogether
Prescription 2
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 100g
Starch 120.0g
Microcrystalline Cellulose 40.0g
The 2%HPMC aqueous solution is an amount of
Magnesium stearate 2.0g
Prepare 1000 altogether
Prescription 3
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 40g
Starch 120.0g
Microcrystalline Cellulose 40.0g
The 2%HPMC aqueous solution is an amount of
Magnesium stearate 2.0g
Prepare 1000 altogether
2, concrete steps:
(1) with Folium Ginkgo extract and Tanshinone I I
AIt is standby that sodium sulfonate was pulverized 100 mesh sieves;
(2) take by weighing supplementary material according to recipe quantity;
(3) hypromellose 2% the aqueous solution made soluble in water is standby;
(4) with Folium Ginkgo extract, Tanshinone I I
ASodium sulfonate, starch, microcrystalline Cellulose mix homogeneously, adding 2%HPMC aqueous solution is an amount of, stirs, and makes suitable soft material;
(5) cross 20 mesh sieve system granules;
(6) granule is dried under 60 ℃ condition;
(7) dry good granule adds magnesium stearate, crosses 18 mesh sieve granulate, mix homogeneously;
(8) sampling, the semi-finished product chemical examination;
(9) loading amount of determining according to chemical examination incapsulates;
(10) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 8 medicinal composition soft capsules of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 40g
Soybean oil 400g
Soybean phospholipid 20g
Cera Flava 12g
Prepare 1000 altogether
Prescription 2
Folium Ginkgo extract 25g
Tanshinone I I
ASodium sulfonate 80g
Soybean oil 400g
Soybean phospholipid 20g
Cera Flava 12g
Prepare 1000 altogether
Prescription 3
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 20g
Soybean oil 400g
Soybean phospholipid 20g
Cera Flava 12g
Prepare 1000 altogether
2, concrete steps:
(1) soybean oil of recipe quantity and soybean phospholipid, Cera Flava heating and melting, mixing is put cold;
(2) add Folium Ginkgo extract and Tanshinone I I
ASodium sulfonate, mixing is crossed colloid mill;
(3) sampling, the semi-finished product chemical examination;
(4) be pressed into soft capsule;
(5) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 9 medicament composition dropping pills of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 40g
Polyethylene glycol 6000 600g
Prescription 2
Folium Ginkgo extract 50g
Tanshinone I I
ASodium sulfonate 80g
Polyethylene glycol 6000 600g
Prescription 3
Folium Ginkgo extract 25g
Tanshinone I I
ASodium sulfonate 60g
Polyethylene glycol 6000 600g
2, concrete steps:
(1) polyethylene glycol 6000 heating and melting in water-bath;
(2) add Folium Ginkgo extract and Tanshinone I I after the whole fusions of polyethylene glycol 6000
ASodium sulfonate, stirring and dissolving;
(3) crossing 60 mesh sieves filters;
(4) maintenance splashes in the liquid paraffin that is chilled to below 10 ℃ for 60 ℃ and makes ball;
(5) finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 10 drug composition oral liquid of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 125g
Tanshinone I I
ASodium sulfonate 100g
Polyoxyethylene sorbitan monoleate 125g
Sodium benzoate 15g
Stevioside 10g
Water adds to 10000ml
Prepare 1000 altogether
Prescription 2
Folium Ginkgo extract 125g
Tanshinone I I
ASodium sulfonate 80g
Polyoxyethylene sorbitan monoleate 125g
Sodium benzoate 15g
Stevioside 10g
Water adds to 10000ml
Prepare 1000 altogether
Prescription 3
Folium Ginkgo extract 100g
Tanshinone I I
ASodium sulfonate 100g
Polyoxyethylene sorbitan monoleate 100g
Sodium benzoate 15g
Stevioside 10g
Water adds to 10000ml
Prepare 1000 altogether
2, concrete steps:
(1) polyoxyethylene sorbitan monoleate is mixed with 20% aqueous solution, with Folium Ginkgo extract and Tanshinone I I
ASodium sulfonate adds the heated and stirred dissolving fully;
(2) sodium benzoate and stevioside is complete with the water dissolution of dosing amount 20%;
(3) merge above-mentioned two solution, mend and add water to full dose;
(4) filtering with microporous membrane of mistake 0.8 μ m;
(5) semi-finished product chemical examination;
(6) fill; Finished product is examined entirely, the packing warehouse-in.
The preparation of embodiment 11 medicament composition granules of the present invention
1, prescription:
Prescription 1
Folium Ginkgo extract 125g
Tanshinone I I
ASodium sulfonate 20g
Icing Sugar 1800.0g
The 2%HPMC50% alcoholic solution is an amount of
Prepare 1000 bags altogether
Prescription 2
Folium Ginkgo extract 75g
Tanshinone I I
ASodium sulfonate 100g
Icing Sugar 1800.0g
The 2%HPMC50% alcoholic solution is an amount of
Prepare 1000 bags altogether
Prescription 3
Folium Ginkgo extract 100g
Tanshinone I I
ASodium sulfonate 40g
Icing Sugar 1800.0g
The 2%HPMC50% alcoholic solution is an amount of
Prepare 1000 bags altogether
Prescription 4
Folium Ginkgo extract 40g
Tanshinone I I
ASodium sulfonate 40g
Icing Sugar 1900.0g
The 2%HPMC50% alcoholic solution is an amount of
Prepare 1000 bags altogether
Prescription 5
Folium Ginkgo extract 40g
Tanshinone I I
ASodium sulfonate 80g
Icing Sugar 1900.0g
The 2%HPMC50% alcoholic solution is an amount of
Prepare 1000 bags altogether
Prescription 6
Folium Ginkgo extract 40g
Tanshinone I I
ASodium sulfonate 120g
Icing Sugar 1900.0g
The 2%HPMC50% alcoholic solution is an amount of
Prepare 1000 bags altogether
2, concrete steps:
(1) it is standby sucrose to be pulverized 100 mesh sieves; With Folium Ginkgo extract and Tanshinone I I
AIt is standby that sodium sulfonate was pulverized 100 mesh sieves;
(2) take by weighing supplementary material according to recipe quantity;
(3) with Folium Ginkgo extract, Tanshinone I I
AThe method mix homogeneously that sodium sulfonate and Icing Sugar progressively increase with equivalent, adding 2%HPMC50% alcoholic solution is an amount of, stirs, and makes suitable soft material;
(4) cross 20 mesh sieve system granules;
(5) granule is dried under 60 ℃ condition;
(6) dried granule is crossed 18 mesh sieve granulate;
(7) sampling, the content of principal agent is determined loading amount in the semi-finished product chemical examination granule;
(8) packing; Finished product is examined entirely, the packing warehouse-in.