CN1172703C - Gingko leaf powder for injection and its prepn. - Google Patents
Gingko leaf powder for injection and its prepn. Download PDFInfo
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- CN1172703C CN1172703C CNB021153590A CN02115359A CN1172703C CN 1172703 C CN1172703 C CN 1172703C CN B021153590 A CNB021153590 A CN B021153590A CN 02115359 A CN02115359 A CN 02115359A CN 1172703 C CN1172703 C CN 1172703C
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Abstract
The present invention discloses a ginkgo leaf powder and injection preparation. The powder and injection preparation can be prepared with the method: ginkgo leaf is extracted through the refluxing of 40 to 80% of ethanol, ethanol is recovered thoroughly through pressure reduction, concentration and standing are carried out, insoluble substances of the upper layer are removed, extraction and recycle are carried out with equivalent ethyl acetate, residues are dried in vacuum and are pulverized into fine powder, fine powder is placed in a supercritical extractor, liquid is extracted with liquid carbon dioxide, and ginkgo leaf extract is obtained. The ginkgo leaf extract is dissolved with water for injection, additives are added for evenly mixing, the pH value is regulated to 6.0 to 7.0, filtration is carried out, filtrate is filtered by a microporous filtration film, filtrate is separately packed in sterilized Xilin bottles in an aseptic mode, and finished products are obtained through freeze-drying, piston pressing and cover pressing. The ginkgo leaf powder and injection preparation of the present invention has the effect of blood activation, clot absorption and meridian passage activation, and has favorable therapeutic effect on apoplexy and apoplexy involving the meridians caused by obstruction of collaterals by blood stasis.
Description
Affiliated technical field
The present invention relates to a kind of gingko leaf powder for injection, the invention still further relates to the preparation method of this gingko leaf powder for injection simultaneously.
Background technology
Apoplexy has another name called apoplexy.The primary disease onset is hurried, and card is seen multiterminal, change rapidly, the feature similarity that becomes with the wind having the nature of shifting number, thus with the apoplexy name it.Chinese medicine is thought: the principal element of its generation is patient's deficiency of qi and blood usually, and with the heart, liver, kidney three dirty imbalance of YIN and YANG, to be worried angry in addition, or drink and be satiated with food, or inducement such as exopathogen invasion and attack, so that QI-blood circulation is obstructed, and skin muscle arteries and veins loses in moistening foster; Or YIN-deficiency in the lower part, liver-yang is opened cruelly, and positive wind-transformation is moving, and bleeding resulting from adverse flow of QI is held expectorant under the arm and is held fire under the arm, and the horizontal channel tunnel of scurrying is hoodwinked key clearly, and forms excess in the upper and deficiency in the lower, the critical syndrome that negative and positive are not maintained mutually.Western medicine thinks that apoplexy is meant " development has the focal obstacle of angiogenic brain function rapidly, and the persistent period surpassed 24 hours or caused the clinical syndrome of death ".Because it makes a general reference breaking out of all cerebrovascular, so claim again: " cerebrovascular accident ".Cerebrovascular is divided into subarachnoid hemorrhage, cerebral hemorrhage, cerebral infarction, cerebral vasospasm etc.The risk factor that causes apoplexy mainly contains hypertension, heart disease, diabetes, hyperlipemia, diseases such as blood-hyperviscosity syndrome.
Apoplexy is one of modal disease of middle-aged and elderly people, has the sickness rate height, fatality rate height, the characteristics that disability rate is high.China adds up in recent years and shows that apoplexy is in second in cause of the death ranking, is only second to malignant tumor.Show that according to domestic Epidemiological study result China's stroke onset rate is 120～1,80/,100,000 people, about 500～6,000,000 people of number of patients.About 1,000,000 people of person that die from the apoplexy every year, survivor about 3/4 is disability in various degree, and severe disabled person accounts for more than 40%, about 130～1,500,000 people of annual neopathy, wherein the case more than 600,000 is a Cerebral Infarction Patients.Therefore, how to improve cerebrovascular, improve patient's life quality, become the important topic of domestic and international medical research at convalescent cure rate.
The method of western medicine treatment cerebral infarction mainly contains following: the cerebral blood flow increasing amount, comprise the outer tremulous pulse bypass Coronary Artery Bypass of intracranial, and can make the mortality rate of apoplexy and recurrence rate reduce 20%.Because cerebral infarction is caused by thrombosis or thromboembolism more than 80%, therefore, the conventional therapy of thrombolytics such as streptokinase and urokinase Chang Zuowei cerebral embolism, the neurological symptom that can improve patient promotes the recovery of function of nervous system, and reduce long-term handicapped generation, but this type of medicine has high intracranial hemorrhage complication; Vasodilation can be expanded the normal blood vessels bed around the hemisphere, but often can not increase the flow of ischemic region blood; Antiplatelet and anticoagulant therapy are verified to be the method that clinical efficacy is arranged, clinical ASP, persantin, the thiophene chloropyridine etc. of mainly containing of antiplatelet drug, and this type of medicine can significantly reduce the relapse rate of apoplexy.Mannitol, furosemide, adrenal gland's glucocorticoid are one of major measure of apoplexy dehydration therapy, use dehydrant energy mitigation symptoms or rescue life rapidly.Calcium ion antagonist, opiate receptor antagonist, platelet activating factor antagonist has the protection therapeutical effect to ischemic brain injury.
The life that above-mentioned treatment is saved patient for acute stage has played positive effect.But doctor trained in Western medicine is heavily treatment often, light rehabilitation, ignored the treatment of patient's convalescent period function, and from the doctor trained in Western medicine angle, at present convalescent period is not still had active and effective Therapeutic Method, the Chinese traditional treatment primary disease is emphasized treating both the principal and secondary aspects of a disease, in conjunction with methods such as the acupuncture of the traditional Chinese medical science, massage, dippings, for the fatality rate that reduces post-stroke, disability rate, relapse rate have played significant effect.At present, be used for the treatment of nearly hundred kinds of the convalescent Chinese herbal medicine of apoplexy, modern pharmacy research and clinical research confirm that all this disease is had definite curative effect, and new drug constantly occurs, but along with being on the rise of China's problem of an aging population, apoplexy patient sharply rise, and the convalescent medicine of treatment apoplexy can not satisfy patient's demand far away.
Summary of the invention
The object of the present invention is to provide a kind of gingko leaf powder for injection that apoplexy has definite curative effect for the treatment of.
Another object of the present invention also is to provide the preparation method of above-mentioned gingko leaf powder for injection.
Gingko leaf powder for injection provided by the invention, prepared by following method: get Folium Ginkgo, with 40～80% alcohol reflux, reclaim under reduced pressure is ethanol to the greatest extent, concentrate, leave standstill, remove the upper strata insoluble matter, with equivalent ethyl acetate extraction and recovery, residue is vacuum drying under 60 ℃ of conditions, being ground into fine powder, fine powder is put in the supercritical extraction device, is fluid extraction with liquid carbon dioxide, get Folium Ginkgo extract, Folium Ginkgo extract is added the dissolving of injection water, add meglumine again, sodium sulfite and mannitol mixing, adjust pH to 6.0～7.5, filter, filtrate is supplied water for injection and regulated pH value once more is 6.0～7.5, the reuse filtering with microporous membrane, and filtrate is aseptic subpackaged in the sterilization cillin bottle, lyophilization, tamponade, gland, promptly.
The invention provides a kind of preparation method of gingko leaf powder for injection, this method comprises the step of following order:
(1) get Folium Ginkgo, with 40～80% alcohol reflux, reclaim under reduced pressure is ethanol to the greatest extent, concentrates, and leaves standstill, and removes the upper strata insoluble matter, also reclaims with the equivalent ethyl acetate extraction, and residue is vacuum drying under 60 ℃ of conditions, is ground into fine powder;
(2) getting above-mentioned fine powder, put in the supercritical extraction device, is fluid extraction with liquid carbon dioxide, gets Folium Ginkgo extract;
(3) get the said extracted thing, add the dissolving of injection water, add meglumine, sodium sulfite and mannitol mixing again, adjust pH to 6.0～7.5, filter, filtrate is supplied water for injection and regulated pH value once more is 6.0～7.5, the reuse filtering with microporous membrane, and filtrate is aseptic subpackaged in the sterilization cillin bottle, lyophilization, tamponade, gland, promptly.
In the method for the invention, being concentrated into density behind the most ethanol of reflux, extract, thing reclaim under reduced pressure in the step (1) is 1.02～1.10; Microporous filter membrane described in the step (3) is respectively 0.65 μ m, 0.22 μ m microporous filter membrane.
Semen Ginkgo has very long medicinal, edible history in China, have the title of living fossil.According to " Treatise on Dietetic Therapy " record, it can be used for severe palpitation, diseases such as cough and asthma due to lung deficiency.Contain flavone compound in the modern medicine study proof Folium Ginkgo, bioactive ingredients such as terpenoid, these compositions have quite intensive antioxidation, can remove free radical superfluous in the organism, stop the body lipid peroxidating, improve the immunity of body etc.Its flavonoid composition, the energy coronary dilating improves the heart, cerebral blood circulation, reduces serum cholesterol, loose bronchus.Nearly recent decades, Folium Ginkgo extract (GBE) has been carried out more deep research both at home and abroad, wherein bilobalide has been put into the important drugs for the treatment of cardiovascular and cerebrovascular disease.Bilobalide is diterpenoid-lactone with special construction and the sesquiterpene lactones of finding from Folium Ginkgo.At present, from Folium Ginkgo, isolated ginkalide A, ginkalide B, ginkalide Cs etc. are the confessed a kind of antagonist that the special effective Platelet of application prospect is arranged most of current the world of medicine, its specificity height.Up-to-date pharmacological experiments proves: Folium Ginkgo terpene lactones has protective effect to ischemia injury and central nervous system; the platelet aggregation of prevention and thrombotic magical function are arranged; in addition; also has antiallergic; effect such as shock, anti-organ transplant rejection is the natural drug of the treatment cardiovascular and cerebrovascular disease of first-selection.
Gingko leaf powder for injection of the present invention has the effect of blood circulation promoting and blood stasis dispelling, dredge the meridian passage, and the card of the apoplexy apoplex involving the channels and collaterals that obstruction of collaterals by blood stasis is caused has excellent curative.
Embodiment 1
Get Folium Ginkgo 5.5kg, add 12 times of amount 70% ethanol, reflux, extract, 2 times, each 1 hour, filter, merge alcohol extract, reclaim under reduced pressure is ethanol to the greatest extent, and to be concentrated into relative density be 1.08 (60 ℃), left standstill 48 hours, remove the upper strata insoluble matter, use equivalent ethyl acetate extraction 7 times, merge extractive liquid,, reclaim ethyl acetate to the greatest extent, residue is ground into fine powder at 60 ℃ of vacuum dryings, fine powder is put in the supercritical extraction device, is fluid extraction with liquid carbon dioxide, gets Folium Ginkgo extract.Get Folium Ginkgo extract, add 4000ml water for injection, stirring makes dissolving, adds meglumine 22g again, sodium sulfite 2.5g, mannitol 70g, mixing with 10% sodium bicarbonate solution adjust pH to 6.0～7.5, filters, filtrate is added water for injection to 5000ml, reuse 10% sodium bicarbonate solution adjust pH to 6.0～7.5 are used 0.65 μ m respectively, 0.22 μ m filtering with microporous membrane, filtrate is aseptic subpackaged in the sterilization cillin bottle, every bottle of 5ml, lyophilization, tamponade, gland promptly gets gingko leaf powder for injection of the present invention.
Embodiment 2
Get Folium Ginkgo 6kg, add 10 times of amount 60% ethanol, reflux, extract, 3 times, each 1 hour, filter, merge alcohol extract, reclaim under reduced pressure is ethanol to the greatest extent, and to be concentrated into relative density be 1.05 (60 ℃), left standstill 72 hours, remove the upper strata insoluble matter, use equivalent ethyl acetate extraction 8 times, merge extractive liquid,, reclaim ethyl acetate to the greatest extent, residue is ground into fine powder at 60 ℃ of vacuum dryings, fine powder is put in the supercritical extraction device, is fluid extraction with liquid carbon dioxide, gets Folium Ginkgo extract.Get Folium Ginkgo extract, add 4200ml water for injection, stir and make dissolving, add meglumine 20g again, sodium sulfite 2.3g, mannitol 75g, mixing with 10% sodium bicarbonate solution adjust pH to 6.0～7.5, filters, filtrate is added water for injection to 5000ml, and reuse 10% sodium bicarbonate solution adjust pH to 6.0～7.5 are used 0.65 μ m respectively, 0.22 μ m filtering with microporous membrane, filtrate is aseptic subpackaged in the sterilization cillin bottle, lyophilization, tamponade, gland promptly gets gingko leaf powder for injection of the present invention.
Pharmacodynamic test of active extract
Gingko leaf powder for injection iv7.416,2.472,0.824g crude drug/kg can significantly reduce the infarction size and the water content of rat cerebral tissue after the Focal Cerebral Ischemia Reperfusion, improve its neurobehavioral.Brain tissue homogenate's biochemical indicator detects finds that this medicine can significantly reduce MDA content in the cerebral tissue, increased SOD content.The cerebral tissue pathological section shows that this medicine is obvious to the neurocyte protection effect.Show that this medicine has significant protective effect to the rat experiment focal cerebral ischemia.Gingko leaf powder for injection iv can obviously shorten the righting reflex recovery time of experimental dispersivity global brain ischemia rat, can also significantly reduce behind the cerebral ischemia re-pouring content of azovan blue in the cerebral tissue.Promptly reduce cerebrovascular permeability, show that this medicine has protective effect to rat experiment dispersivity global brain ischemia.
Gingko leaf powder for injection iv can obviously increase the survival number in the 2h after the cerebral ischemia of chmice acute imperfection, shows that cerebral ischemia has protective effect, the ED of iv to this medicine to the chmice acute imperfection
50Be 1.90g/kg (the 95% credible 1.43-2.53g/kg that is limited to; The ED of ig
50Be 3.25g/kg (the 95% credible 2.68-51.7g/kg that is limited to).10min promptly begins onset behind the Iv, and about peak time 20min, the persistent period can reach more than the 90min; And ig onset time 20-30min, about peak time 45min, the persistent period can reach more than the 120min.The effect of same dosage group iv is better than ig.Can also obviously prolong the time-to-live of mice under anaerobic condition, show that this can improve the ability of mice anoxia enduring.
Gingko leaf powder for injection iv administration is to the cerebral blood flow of anesthetized dog, cerebral vascular resistance, and blood pressure, heart rate and electrocardio all do not have obvious influence.
Gingko leaf powder for injection induces the rabbit extracorporeal platelet aggregation that obvious inhibitory action is arranged to PAF.A little less than inducing rabbit extracorporeal platelet aggregation inhibitory action to ADP.This medicine iv induces to PAF that platelet aggregation also has obvious inhibitory action in the rabbit body; This medicine does not have obvious influence to platelet.Can reduce rabbit erythrocyte hematocrit and whole blood viscosity during iv, especially more obvious to the whole blood viscosity improvement effect of high shearing, high dose group during to low, high shearing whole blood viscosity the improvement effect is all arranged.Iv can obviously suppress rat and rabbit moves-vein bypass thrombosis.Iv can obviously prolong clotting time of mice.
The test of general pharmacology research
Under domesticated dog anesthesia (pentobarbital sodium i.v30mg/kg) state, large, medium and small three dosage of intravenous drip gingko leaf powder for injection (2.7,0.9,0.3g/kg crude drug three dosage), cumulative volume 10ml/kg, the blood pressure (SBP, DBP, BP) that administration 5,10,15,30,60,90,120 minutes is measured, respiratory frequency, amplitude, body temperature, heart rate, electrocardiogram (P, QRS, ST, T ripple interval, amplitude) parameter has no significant change.
Large, medium and small three the dosage groups of gingko leaf powder for injection (9.0,4.5,1.5g/kg crude drug three dosage), press the injection of 0.1ml/10g body weight mouse peritoneal, behind the medicine 30 minutes, to mouse peritoneal injection 45mg/kg pentobarbital sodium, the length of one's sleep of animal and normal saline matched group be no difference of science of statistics (P＞0.05) relatively all; The heavy dose of group of the mouse peritoneal injection 25mg/kg sleeping number of pentobarbital sodium animal relatively there is significant difference (P＜0.05) with matched group; To spontaneous activity number of times in the mice 10 minutes, big or middle dosage group and matched group be P＜0.05 relatively.
In sum: the above-mentioned dosage of gingko leaf powder for injection does not have influence to dog blood pressure, breathing, body temperature, electrocardiogram; High dose and pentobarbital sodium have synergism, can increase the sleeping number of mice; The mice autonomic activities there is certain inhibitory action.
Animal acute toxicity test
The gingko leaf powder for injection mouse mainline, the maximum dosage-feeding mouse death rate is 30%, the iv acute toxicity test can't be measured LD
50, mice iv maximum tolerated dose is 21.3g crude drug/kg, 128 times of the clinical application that is equivalent to (clinical plan consumption is 10g crude drug/60kg/ day, i.e. 0.167g crude drug/kg).Gingko leaf powder for injection mouse peritoneal injection maximum dosage-feeding mouse death rate is 15%, and the ip administration can't LD
50, show gingko leaf powder for injection mouse peritoneal injection LD through result of the test
50Greater than 53.4g crude drug/kg, be equivalent to 320 times of clinical plan amount.
Long-term toxicity test for animals
1, rat long term toxicity test
Get 80 of Sprague-Dawley rats, that test divides is high (600mg extract/kg), in (190mg extract/kg), low (60mg extract/kg) dosage group and 5% glucose injection matched group, 20 of every treated animals, male and female half and half.Lumbar injection, once a day, administration is 6 days weekly, and the test period was 13 weeks.Each test group stays 1/2 animal to change to observe for 4 convalescent periods in week.The result shows that the behavioral activity of rat, fur color and luster and feces all belong to normally.Food ration is not found the significantly ANOMALOUS VARIATIONS relevant with drug effect.The body weight gain of high, medium and low dosage treated animal is compared no significant difference with matched group.Main organ weights and coefficient thereof and matched group be unknown significance difference relatively.Hematology and blood biochemical are learned experiment and are shown, every index that high, medium and low dosage group rat blood is detected and matched group do not see obviously because the ANOMALOUS VARIATIONS of drug-induced.The pathological section result shows, indivedual main organs all have MC in various degree, analyze the caused pathological change of non-medicine itself by Ridit, the change of lungs may be due to virus or the bacterial infection, should note the variation of liver index in clinical trial.Infer that in view of the above the continuous lumbar injection of high, normal, basic dosage gives this medicine 13 weeks, rat does not show tangible toxic reaction.Under this experimental condition, continuous 13 weeks of ip administration of this medicine are 600mg extract/kg to the non-toxic of rat.If the people is in 60kg, (600mg extract/kg) is equivalent to 120 times (human dosage is 300mg extract/sky/60kg people) of human dosage to gingko injection approximately to the non-toxic of rat.
2, Canis familiaris L. long term toxicity test
The various reactions that this medicine continuous intravenous injection of this experimental observation is (iv) taken place the Beagle dog in 13 weeks of administration are to assess the safety of this medicine.24 of Beagle dogs, test divide a medicine height (300mg extract/kg), in (95mg extract/kg), low (30mg extract/kg) group and blank group (5% glucose injection).6 of every treated animals, male and female half and half.Intravenous injection, once a day, 6 times weekly, the test period was 13 weeks.Each test group is prepared 2 animals and is observed 4 convalescent periods in week variations.The result shows that the significantly ANOMALOUS VARIATIONS relevant with drug effect all found to routine blood test, serum biochemistry, routine urinalysis and the food ration etc. of dog in these medicine 13 all backs of intravenous injection.All are normal after administration for the activity situation of animal.Compare with matched group, the dog body weight of each administration group and body weight gain normal and in week no significant difference.The every index of dog electrocardiogram is normal.The tissue slice inspection shows, examined at all to there is no tangible pathological change in the internal organs.Under this experimental condition, 13 weeks of gingko leaf powder for injection iv administration the non-toxic to dog be 300mg extract/kg, be equivalent to 60 times (human dosage is 300mg extract/sky/60kg people) of human dosage approximately.
Safety testing
Gingko leaf powder for injection does not have haemolysis and red cell agglutination to Beagle dog limb venous blood, and healthy guinea pig and Beagle dog are not had irritated reaction, and the rabbit intramuscular injection is not seen that the injection site muscle groups is woven with abnormal change.To the no obvious vascular stimulation reaction of tame rabbit ear vein instillation, rabbit there is not pyrogen reaction.Through above test, the result shows that said preparation is safe.
Claims (4)
1, a kind of gingko leaf powder for injection, this injectable powder is prepared by following method: get Folium Ginkgo, with 40～80% alcohol reflux, reclaim under reduced pressure is ethanol to the greatest extent, concentrate, leave standstill, remove the upper strata insoluble matter, with equivalent ethyl acetate extraction and recovery, residue is vacuum drying under 60 ℃ of conditions, being ground into fine powder, fine powder is put in the supercritical extraction device, is fluid extraction with liquid carbon dioxide, get Folium Ginkgo extract, Folium Ginkgo extract is added the dissolving of injection water, add meglumine again, sodium sulfite and mannitol mixing, adjust pH to 6.0～7.5, filter, filtrate is supplied water for injection and regulated pH value once more is 6.0～7.5, the reuse filtering with microporous membrane, and filtrate is aseptic subpackaged in the sterilization cillin bottle, lyophilization, tamponade, gland, promptly.
2, a kind of preparation method of gingko leaf powder for injection, this method comprises the step of following order:
(1) get Folium Ginkgo, with 40～80% alcohol reflux, reclaim under reduced pressure is ethanol to the greatest extent, concentrates, and leaves standstill, and removes the upper strata insoluble matter, also reclaims with the equivalent ethyl acetate extraction, and residue is vacuum drying under 60 ℃ of conditions, is ground into fine powder;
(2) getting above-mentioned fine powder, put in the supercritical extraction device, is fluid extraction with liquid carbon dioxide, gets Folium Ginkgo extract;
(3) get the said extracted thing, add the dissolving of injection water, add meglumine, sodium sulfite and mannitol mixing again, adjust pH to 6.0～7.5, filter, filtrate is supplied water for injection and regulated pH value once more is 6.0～7.5, the reuse filtering with microporous membrane, and filtrate is aseptic subpackaged in the sterilization cillin bottle, lyophilization, tamponade, gland, promptly.
3, the preparation method of gingko leaf powder for injection according to claim 2 is characterized in that: being concentrated into density behind the most ethanol of reflux, extract, thing reclaim under reduced pressure in the step (1) is 1.02～1.10.
4, the preparation method of gingko leaf powder for injection according to claim 2 is characterized in that: the microporous filter membrane described in the step (3) is respectively 0.65 μ m, 0.22 μ m microporous filter membrane.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB021153590A CN1172703C (en) | 2002-06-11 | 2002-06-11 | Gingko leaf powder for injection and its prepn. |
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| CNB021153590A CN1172703C (en) | 2002-06-11 | 2002-06-11 | Gingko leaf powder for injection and its prepn. |
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| CN1389240A CN1389240A (en) | 2003-01-08 |
| CN1172703C true CN1172703C (en) | 2004-10-27 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101890061A (en) * | 2010-07-16 | 2010-11-24 | 悦康药业集团有限公司 | Method for preparing ginkgo biloba leaf extract injection, and ginkgo biloba leaf extract injection |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111719113B (en) * | 2019-03-19 | 2021-08-17 | 中山大学 | Surface modification method of acupuncture needle |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101890061A (en) * | 2010-07-16 | 2010-11-24 | 悦康药业集团有限公司 | Method for preparing ginkgo biloba leaf extract injection, and ginkgo biloba leaf extract injection |
| CN101890061B (en) * | 2010-07-16 | 2012-04-18 | 悦康药业集团有限公司 | Method for preparing ginkgo biloba leaf extract injection, and ginkgo biloba leaf extract injection |
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