CN100496520C - Chinese-medicinal extract for treating cardiovascular disease, its preparation and use - Google Patents
Chinese-medicinal extract for treating cardiovascular disease, its preparation and use Download PDFInfo
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- CN100496520C CN100496520C CNB2006100857392A CN200610085739A CN100496520C CN 100496520 C CN100496520 C CN 100496520C CN B2006100857392 A CNB2006100857392 A CN B2006100857392A CN 200610085739 A CN200610085739 A CN 200610085739A CN 100496520 C CN100496520 C CN 100496520C
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Abstract
An extract of Chinese-medicinal material for treating cardiovascular and cerebrovascular diseases, coronary heart disease and cerebral stroke is prepared from lucid ligustrum fruit. Its preparing process is also disclosed.
Description
Technical field
The present invention relates to natural medicine field, specifically disclose extract, its preparation method of Fructus Ligustri Lucidi, the pharmaceutical composition that contains it and medical usage thereof.The extract of Fructus Ligustri Lucidi of the present invention can be used for cardiovascular and cerebrovascular disease, particularly coronary heart disease and apoplexy treatment.
Background technology
Fructus Ligustri Lucidi is the dry mature fruit of Oleaceae plant Fructus Ligustri Lucidi (Ligustrum Lucidum Ait.), is the conventional Chinese medicine of strengthening the body resistance.Chinese medicine circle thinks that always its effective ingredient is oleanolic acid and polysaccharide, and has carried out a lot of studies of pharmacy.The someone has reported and has contained rhodioside, aglycon in the Fructus Ligustri Lucidi successively in recent years, and has carried out analysis and assay, thinks to have only the saying that contains rhodioside in the Radix Rhodiolae thereby broken people always.But regrettably, nobody carries out the research that rhodioside and effective site thereof are extracted in scale from Radix Rhodiolae, and also nobody carries out the pharmacodynamic study of this extract, thereby Radix Rhodiolae be not applied aspect the treatment cardiovascular and cerebrovascular disease.
Summary of the invention
One of purpose of the present invention provides extract of a kind of Fructus Ligustri Lucidi and preparation method thereof.
Another object of the present invention is to disclose a kind of pharmaceutical composition that contains this extract.
A further object of the invention is the therapeutic use that discloses this compositions, promptly treats the purposes of cardiovascular and cerebrovascular disease.
Extract of the present invention is prepared by following method: as extracting the solvent extraction Fructus Ligustri Lucidi, extracting solution is concentrated into extractum with water-soluble solvent, drying, and extracting solution is concentrated into to be done or is concentrated in right amount, obtains extracting concentrate.Described water-soluble solvent is selected from water, acetone or C
1-4The aliphatic alcohol of straight or branched in the mixed solvent of one or more solvent compositions.The extraction concentrate that obtains obtains extract through organic solvent extraction, and drying promptly gets crude extract.Described organic solvent is selected from one or more of petroleum ether, chloroform, dichloromethane, ethyl acetate, n-butyl alcohol.
Above-mentioned preparation method further can comprise: the crude extract that obtains separates with chromatographic column, ethanol with volume ratio 0-100:100-0: water or methanol: water or acetone: water carries out gradient elution, the Fractional Collections effluent, wherein said chromatographic column is selected from macroporous resin column, polyamide column, silicagel column, alumina column or sephadex column; Will contain in the above-mentioned effluent effluent of rhodioside and butyl alcohol merge, reclaim solvent, be concentrated into driedly, obtain extract after the drying.
Contain rhodioside and butyl alcohol among the present invention.General, the amount that contains rhodioside and butyl alcohol in the crude extract is 3-15%, and the amount in the extract is 30-60%, and therefore, quality control selects the content of rhodioside and butyl alcohol as a quality index.
The present composition also discloses a kind of pharmaceutical composition, and it contains the Fructus Ligustri Lucidi extract and the pharmaceutically acceptable carrier for the treatment of effective dose.
Described Fructus Ligustri Lucidi extract can be a crude extract, also can be extract.Can be dosage forms such as tablet, capsule, pill, powder, powder, granule, lozenge, dispersible tablet, suppository, soft capsule, oral liquid, suspensoid, big or small-volume injection, aseptic parenteral solution, freeze-dried powder, aseptic powder packing.
The dosage of above activating agent will be different because of prescription.General dosage is to contain extract 3-2000mg in the per unit preparation, also can be according to different dosage form selection different dosage.
The present invention can be used for cardiovascular and cerebrovascular disease, is preferably the treatment of coronary heart disease and apoplexy.In pharmacological testing, discovery this product can be improved the hemodynamic situation of heart, alleviates the treating myocardial ischemia damage degree, dwindles the treating myocardial ischemia damage scope.This product also has significant protective effect to the damage of rat cerebral infarction model, and the focus area is significantly dwindled, and tissue water content reduces, and the edema degree obviously alleviates, and obviously improves the hypoxia-bearing capability of body.
Below be part of test results of the present invention:
Test 1: the present composition is to the influence of anesthesiaing dog heart blood flowing dynamics
Test drug be according to the embodiment of the invention 1 preparation extract, be divided into two dosage groups: small dose group (20mg.kg
-1) and heavy dose of group (60mg.kg
-1).The contrast medicine is Radix Notoginseng total arasaponins tablet (hereinafter to be referred as the total glycosides sheet).
It is 24 of the healthy hybrid dogs of 8～12kg that experimental animal is selected body weight for use, male and female half and half, female unpregnancy.With distilled water extract and Radix Notoginseng total arasaponins tablet are made into test desired concn gastric infusion, are referred to as extract group and total glycosides sheet group respectively, and the test group that will only give the dosage solvent is defined as the solvent matched group.
Experimental condition is 18～22 ℃ of room temperatures (air-conditioning control), humidity 55～65%.Each dog is got dorsal position and fixes under 3% Nembutal vein anesthetic.The cervical region median incision exposes and tracheostomize, and the trachea intubate is in order to the artificial respiration; Separate the left side jugular vein and carry out oxygen content mensuration in order to the coronary sinus vein intubate; Operation separates the left side femoral artery, and water receiving silver manometer is directly traced blood pressure; Separate right femoral artery and carry out arterial oxygen content mensuration in order to blood sampling; Separate left femoral vein and connect infusion bottle, to replenish the body fluid of losing; The subcutaneous fixedly needle electrode of extremity is traced the electrocardiogram that standard limbs II leads, to calculate heart rate; Animal is got the right clinostatism of forelimb then, the anterior pectorial region unhairing, Yu Zuosi, five intercostals cut skin along the 4th rib lower edge, the passivity separating muscle, after exposing pleura, connect artificial respirator, fully pleura is cut in the hemostasis back, mention pericardium and cut and do the pericardium bed, stay preplaced line behind careful separation LCA and the aortic root,, select suitable electromagnetic flowmeter probe to debug according to coronary artery and aortal thickness; (inject anticoagulant heparin in the cardiac catheter) from right jugular vein intubate to coronary sinus vein; Coronary artery and aorta are by last suitable electromagnetic flowmeter probe.The abdominal part median incision is found out duodenum in order to administration.After aforesaid operations finished, the intravenous injection heparin carried out whole body heparinization (5mg/kg).After the heparinization, after inspection has or not hemorrhage and abundant hemostasis once more, stablized 30 minutes.Record and recording blood pressure, electrocardiogram and coronary artery, aorta flow, femoral artery and coronary sinus vein are got blood and are carried out blood gas analysis etc., as the normal value before the administration.Behind different groups and dosed administration, after administration, measured coronary sinus vein, ABF, blood pressure and electrocardiogram etc. respectively in 20,40,60,120 and 150 minutes again, and coronary sinus vein and aortic blood oxygen content are surveyed in 40 minutes and blood sampling in 60 minutes after administration.After experiment finishes, take out heart and weigh.
The measurement result of blood pressure, heart rate, coronary artery blood flow and the oxygen content, ABF and the oxygen content that obtain is carried out statistical procedures, and calculate coronary artery blood flow, cardiac output, per minute hectogram myocardial flow, coronary resistance etc., and its statistical procedures result is recorded in table 1～3 respectively according to formula.
Table 1. extract is to the influence (n=6) of dog coronary flow (ml/min), cardiac output (L/min)
With before the administration relatively: * P＜0.05 * * P＜0.01 and total saponins sheet group relatively: △ P＜0.05 △ △ P＜0.01
As seen the extract group has obviously improved dog coronary flow and cardiac output after administration, and the heavy dose of group of extract is significantly improved than the coronary flow of total saponins sheet group in the time of 60 minutes.
Table 2. extract is to the influence (n=6) of dog coronary resistance (mmHg/ml/100g/min)
With before the administration relatively: * P＜0.05 * * P＜0.01 and total saponins sheet group relatively: △ P＜0.05
As seen the extract group has obviously reduced the dog coronary resistance after administration, and the heavy dose of group of extract is obvious than total saponins sheet group effect in the time of 120 minutes.
Table 3. extract is to the influence (n=6) of dog per minute hectogram myocardial flow (ml/100g/min)
With before the administration relatively: * P＜0.05 * * P＜0.01 and total saponins sheet group relatively: △ P＜0.05 △ △ P＜0.01
As seen the extract group has obviously reduced the dog coronary resistance after administration, and the extract group is obvious than total saponins sheet group effect in the time of 60 minutes.
Test 2: to the protection of the dead damage of real face rat heart infarction
Get 50 of wistar rats, body weight 250～300g, male and female half and half, be divided into sham operated rats (waiting capacity 0.5%CMCNa) at random, model group (waiting capacity 0.5%CMCNa), total glycosides sheet group (50mg/kg, calculate by kg body weight, be equivalent to 10 times of clinical people's consumption 168mg/d), extract little (20mg/kg) and heavy dose of group (60mg/kg), behind the successive administration 3 days, rat is used etherization, open breast in a left side the 4th intercostal and expose heart, apart from left coronary artery starting point 2～3mm place, wear 50 surgical thread ligation, close breast, sham operated rats is only opened breast, not ligation coronary artery.0.5,1,2,3,5 hour electrocardiogram is measured in administration immediately after operation finishes, and observes the ST section value of raising, and the results are shown in Table 4.Perform the operation and put to death rat in back 24 hours, open breast immediately, take out heart,, at the bottom of the heart, get four altogether from the apex of the heart from coronary sulcus excision atrium, every thick about 0.1cm, with the 0.5%NBT dyeing liquor 15min that in 37 ℃ of thermostat water baths, dyes, separate infarcted region, weigh, calculate the percentage ratio that infarcted region weight accounts for ventricular weight, the results are shown in Table 5.
Table 4. extract is to the influence of rat model ECG ST section (x ± s)
With model group than * p＜0.05 * * p＜0.01, with sham operated rats than △ △ p＜0.01
Table 5. extract to the rat model myocardial infarction after the influence (x ± s) of myocardial infarction weight
With model group than * p＜0.05 * * p＜0.01, with sham operated rats than △ p＜0.05 △ △ p＜0.01
Test the protection of 3. pairs of real face rat cerebral infarction damages
The SD rat is (male, body weight 300～350g) is anaesthetized with pentobarbital sodium 40mg/kgiP, and the mid point between right eye and auris dextra is made the IJ that cuts of 1.5cm length then, separates clever flesh, expose zygomatic process and clever bone, head end 1～2mm place in zygomatic process opens the aperture that a diameter is 2mm with dental burr, and pia mater encephali is opened an aperture, expose the initial position of mesencephalic arteries, with scraper cerebral tissue is removed gently,, then temporalis and skin layering are sewed up with the bright disconnected middle cerebral artery of small bipolar electrocoagulator.Get 32 rats of performing the operation successfully, be divided into 4 groups: model control group, extract is low, high dose group (20,60mg/kg), NAOXUESHUAN PIAN matched group (0.15g/kg), behind the continuous use 15d,, get its brain with sacrifice of animal, with equal equidistant 5 sections of crosscut, behind 37 ℃ of dyeing 10min, separate infarcted region and non-infarcted region with blue (TTG) phosphate buffer of 1% 4 nitrogen, weigh, calculate infarct size with gravimetric method, at last cerebral tissue is put continuous oven drying 24h in the people I10 ℃ baking oven, taken out title cerebral tissue dry weight, calculate brain water content.The results are shown in Table 6.
Table 6. extract is to the protective effect of rat experiment cerebral infarction (x ± s)
With model group than * p＜0.05 * * p＜0.01
Test the anoxybiotic protection of 4. pairs of real face rat brains
(male, 300～350g) 30 of body weight are divided into 3 groups: matched group (distilled water), extract is low, (20,60mg/kg), after medication 1 hour, the sodium nitrite solution of lumbar injection 3% write down its death time to high dose group for the SD rat.The results are shown in Table 7.
Table 7. extract is to the anoxybiotic protective effect of rat brain (x ± s)
With matched group than * p＜0.05 * * p＜0.01
By the following specific embodiments, further to be well understood to the present invention.
The specific embodiment
Embodiment 1:
Get Fructus Ligustri Lucidi 1kg, add 70% alcohol reflux 2 hours, totally 2 times, merge medicinal liquid, concentrate, reclaim ethanol to relative density 1.2.Add ethyl acetate, n-butanol extraction 2 times respectively, be concentrated to extractum.Extractum adds 10 times of water gaging dissolvings, and the macroporous resin chromatographic column of flowing through separates, and earlier with water elution, 20: 80 ethanol of reuse volume ratio: water carries out the tonsure eluting, the Fractional Collections effluent, and effluent is through concentrating, and the vacuum decompression drying obtains yellow solid.Wherein contain rhodioside 38%, butyl alcohol 4%.
Embodiment 2:
Get Fructus Ligustri Lucidi 1kg, add 60% alcohol reflux 2 hours, totally 2 times, merge medicinal liquid, concentrate, reclaim ethanol to relative density 1.2.Add ethyl acetate, n-butanol extraction 3 times respectively, be concentrated to extractum.Extractum adds 10 times of water gaging dissolvings, and the macroporous resin chromatographic column of flowing through separates, and earlier with water elution, 10: 90 ethanol of reuse volume ratio: water carries out eluting, collects effluent, and effluent is through concentrating, and the vacuum decompression drying obtains faint yellow solid.Wherein contain rhodioside 47%, butyl alcohol 8%.
Embodiment 3:
Get Fructus Ligustri Lucidi 1kg, add 60% alcohol reflux 2 hours, totally 2 times, merge medicinal liquid, concentrate, reclaim ethanol to relative density 1.2.Add ethyl acetate, n-butanol extraction 1 time respectively, be concentrated to extractum.Extractum adds 8 times of water gaging dissolvings, and the silicagel column of flowing through separates, and earlier with water elution, 10: 90 ethanol of reuse volume ratio: water carries out eluting, collects effluent, and effluent is through concentrating, and spray drying obtains yellow powder.Wherein contain rhodioside 18%, butyl alcohol 2%.
Embodiment 4:
Get Fructus Ligustri Lucidi 1kg, add 60% alcohol reflux 2 hours, totally 2 times, merge medicinal liquid, concentrate, reclaim ethanol to relative density 1.2.Add ethyl acetate, n-butanol extraction 3 times respectively, extractum adds 8 times of water gaging dissolvings, and the alumina column of flowing through separates, with water elution, 30: 70 ethanol of reuse volume ratio: water carries out eluting earlier, collects effluent, effluent is through concentrating, and spray drying obtains yellow powder.Wherein contain rhodioside 18%, butyl alcohol 2%.
Embodiment 5:
Get Fructus Ligustri Lucidi extract 2g, add starch 10g, microcrystalline Cellulose 10g, polyvidone 3g adds water system soft material, and cross 20 orders and granulate, 60 ℃ of dryings 2 hours, 24 order granulate add the 0.4g magnesium stearate, mixing, tabletting promptly gets 100.
Embodiment 6:
Get in granule filling to 1 capsule among the embodiment 5, get final product.
Embodiment 7:
Get Fructus Ligustri Lucidi crude extract 200g,, add water system soft material, cross 10 orders and granulate with sucrose 300g, 60 ℃ of dryings 2 hours, 14 order granulate, mixing, fill promptly gets 100 bags.
Embodiment 8:
Get Fructus Ligustri Lucidi extract 10g, add water 500ml, pH value is regulated in dissolving, adds active carbon and stirs and filtered in 20 minutes, boils 15 minutes in 100 ℃ after the embedding, promptly gets 100.
Embodiment 9:
Get Fructus Ligustri Lucidi extract 10g, mannitol 40g adds water 200ml, and pH value is regulated in dissolving, and add active carbon and stir filtration in 20 minutes, with 0.2 μ m microporous filter membrane fine straining, bottling, lyophilization promptly gets 100.
Claims (9)
1, a kind of Fructus Ligustri Lucidi extract, the content sum that it is characterized in that rhodioside and butyl alcohol is 3% to 100%.
2, extract as claimed in claim 1 is prepared by following method: as extracting the solvent extraction Fructus Ligustri Lucidi, extracting solution is concentrated into extractum with water-soluble solvent, drying, and extracting solution is concentrated into to be done or is concentrated in right amount, obtains extracting concentrate; Described water-soluble solvent is water, acetone or C
1-4The aliphatic alcohol of straight or branched in the mixed solvent of one or more solvent compositions; The extraction concentrate that obtains obtains extract through organic solvent extraction, and drying promptly gets crude extract; Described organic solvent is one or more of petroleum ether, chloroform, dichloromethane, ethyl acetate or n-butyl alcohol.
3, claim 1 or 2 described Fructus Ligustri Lucidi extracts, it is characterized in that: described method also comprises: the crude extract that obtains separates with chromatographic column, ethanol with volume ratio 0-100:100-0: water or methanol: water or acetone: water carries out gradient elution, Fractional Collections effluent, wherein said chromatographic column are macroporous resin column, polyamide column, silicagel column, alumina column or sephadex column; Will contain in the above-mentioned effluent effluent of rhodioside and butyl alcohol merge, reclaim solvent, be concentrated into driedly, obtain extract after the drying.
4, any described Fructus Ligustri Lucidi extract of claim 1～3 is used to prepare the purposes of the medicine for the treatment of cardiovascular and cerebrovascular disease.
5, the purposes in the claim 4 is characterized in that: described cardiovascular disease is a coronary heart disease, and described cerebrovascular disease is cerebral ischemia or anoxia.
6, a kind of pharmaceutical composition that is used for the treatment of cardiovascular and cerebrovascular disease is characterized in that: described compositions is made up of any described Fructus Ligustri Lucidi extract and pharmaceutically acceptable carrier in the claim 1～3.
7, according to the pharmaceutical composition described in the claim 6, it is characterized in that: the dosage form of described compositions is tablet, capsule, pill, powder, powder, granule, lozenge, dispersible tablet, suppository, soft capsule, oral liquid, suspensoid, big or small-volume injection, aseptic parenteral solution, freeze-dried powder, aseptic powder packing or suitable sustained-release preparation.
8, pharmaceutical composition according to claim 7 is characterized in that: the dosage form of described compositions is tablet, big or small-volume injection, capsule or freeze-dried powder.
9, pharmaceutical composition according to claim 8 is characterized in that: the dosage form of described compositions is freeze-dried powder or big or little injection.
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|---|---|---|---|
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2013173972A1 (en) * | 2012-05-22 | 2013-11-28 | Nestec S.A. | Fructus ligustri lucidi for growth in children |
| CN104496760B (en) * | 2015-01-06 | 2016-04-27 | 康美（北京）药物研究院有限公司 | A kind of preparation method of high purity tyrosol |
| CN108096380A (en) * | 2018-01-12 | 2018-06-01 | 广西天昌投资有限公司 | A kind of pharmaceutical composition with cardiovascular protective effect and preparation method thereof |
| CN109303786A (en) * | 2018-11-27 | 2019-02-05 | 山东省药学科学院 | A kind of composition of resist oxygen lack and its application |
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