CN101822731A - Raspberry leaf extract and application thereof in preparing anticoagulant and antithrombotic medicine - Google Patents
Raspberry leaf extract and application thereof in preparing anticoagulant and antithrombotic medicine Download PDFInfo
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- CN101822731A CN101822731A CN200910010568A CN200910010568A CN101822731A CN 101822731 A CN101822731 A CN 101822731A CN 200910010568 A CN200910010568 A CN 200910010568A CN 200910010568 A CN200910010568 A CN 200910010568A CN 101822731 A CN101822731 A CN 101822731A
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Abstract
The invention relates to biological medicine, in particular to a raspberry leaf extract and an application thereof in preparing an anticoagulant and antithrombotic medicine, the extract is raspberry leaf water extract, and the obtained raspberry leaf activity extraction part has anticoagulant and antithrombotic activity. The raspberry leaf extract and the application thereof have simple production process, convenient operation and low production cost.
Description
Technical field
The present invention relates to biological medicine, specifically a kind of Fructus Rubi leaf water or pure water extract and this extract are used for anticoagulation and antithrombotic novel medical use.
Background technology
Thrombosis is the surperficial formed fritter of blood flow at inner face exfoliation of cardiovascular system blood vessel or mend.In variable fluid dependent form (variable flow dependent patterns), thrombosis is by insoluble fibrin, sedimentary platelet, leukocyte that gathers and the erythrocyte that is absorbed in are formed. exist the blood coagulation system and the anticoagulation system (fibrinolytic system) of mutual antagonism in the blood.The dynamic equilibrium of blood coagulation system and fibrinolytic system has guaranteed that promptly blood has potential solidifiable to guarantee the fluid state of blood again all the time.Yet, can promote to have broken above-mentioned dynamic equilibrium under the factor effect of coagulation process at some sometimes, triggered coagulation process, blood just can solidify at the cardiovascular intracavity, forms thrombosis.
Thrombosis sickness rate height, harm more very.Statistics, the death that causes because of thrombotic disease have accounted for 51% of the total death toll in the whole world, the death that causes considerably beyond tumor communicate illness, respiratory system disease etc.In every year, China is that the death toll that main apoplexy that shows and myocardial infarction are caused is 2,600,000 with the thromboembolism, and average per 12 seconds with regard to dead 1 people.Thrombotic disease is one of disease of serious harm human health, also is the focus of countries in the world medicine scholar research.From natural product, seek new active component, and and then to develop new medicine also be the important channel of capturing clinical persistent ailment now, the present invention uses up-to-date technological means, in conjunction with modern isolation and identification method, chemical constituent by the Fructus Rubi leaf being carried out system and activity research thereof are in the hope of providing scientific basis for the medicine of seeking new antithrombotic disease.
Red raspberry is referred to as Fructus Rubi again in Chinese materia medica, Fructus Rubi has the effect of tonifying the kidney to consolidate the essence, supporing yang reducing urination, is used for cards such as emission due to deficiency of the kidney, spermatorrhea, sexual impotence, the enuresis, frequent micturition, is warming the kidney to activate YANG Chinese medicine commonly used.Also can be used for dietetic therapy and health care.Pharmacological evaluation proves that Fructus Rubi has antimutagenic, improves ability of learning and memory, slow down aging, the effect of enhance immunity (Xie Yihui etc., Fructus Rubi The Chemical Constituents, Chinese crude drug 2005, Vol.21 (2): 99-100).
The home and abroad is more to report, the research of Fructus Rubi fruit at present, and less to the research of its leaf.The present invention has developed the Folium Rubi chemical constituent has been studied.In addition, Fructus Rubi cures mainly cards such as emission due to deficiency of the kidney, spermatorrhea, sexual impotence, the enuresis, frequent micturition.Do not appear in the newspapers in its effect research to blood coagulation system and thrombus model in the home and abroad at present.
Summary of the invention
In order to meet clinical needs, address the above problem, the purpose of this invention is to provide a kind of Framberry leaf extract and the application in preparation anticoagulation and antithrombotic reagent thereof; Framberry leaf extract can be used as the medicine that active ingredient is used for preparing anticoagulation and anti thrombotic action, promptly can be used for preventing and/or treating in the medicine of diseases such as coronary heart disease, angina pectoris, cerebral thrombosis or pulmonary infarction.
For achieving the above object, the technical solution used in the present invention is:
Framberry leaf extract, it is the water of Fructus Rubi leaf or the extract of pure water.
The water of described Fructus Rubi leaf or the concrete preparation process of the extract of pure water are:
1) 5-25 water or the volume content doubly with Fructus Rubi leaf and Fructus Rubi leaf volume mixes at the alcohol-water solution of 1%-70%, once or once, filters collection filtrate in 60-100 ℃ of lixiviate or reflux, extract; The extraction time of each lixiviate is generally 1-24 hour;
Described alcohol is the full monohydric alcohol CnH that closes with following general formula
2n+1OH, n are 1,2,3 or 4, or monohydric alcohol CnH is closed in insatiable hunger
2n-1OH, n are 2,3 or 4.
2) the Fructus Rubi leaf water of collection or the filtrate of pure water extract are passed through macroporous adsorbent resin, adopt water-alcohol solvent to carry out gradient elution, the volume content of alcohol changes from 0-95% in the eluting solvent, collect the shared eluate of volumn concentration under the institute of the eluting solvent between 5%-95% eluting of alcohol in water-pure mixed solvent, eluate is reclaimed solvent through concentrating under reduced pressure;
Described alcohol is the full monohydric alcohol CnH that closes with following general formula
2n+1OH, n are 1,2,3 or 4, or monohydric alcohol CnH is closed in insatiable hunger
2n-1OH, n are 2,3 or 4.
3) described eluate can adopt concentrate drying, lyophilization or spray drying process drying to be pressed powder after concentrating under reduced pressure reclaims solvent, obtains having the plant extract of anticoagulation and antithrombotic acitivity.
Described plant extract is used to prepare anticoagulation and/or antithrombotic reagent, anticoagulation and/or antithrombotic preventive health care's food or the food additive that prevents and/or treats to purpose.
Described preparation prevents and/or treats anticoagulation and/or antithrombotic medicine and is meant its made pharmaceutical dosage form of claim 1 be can be pill, powder, tablet, unguentum, powder, injection, water preparation, granule or injection.
Described preventing and/or treating to the anticoagulation of purpose and/or antithrombotic reagent are that described plant extract is combined as excipient as acceptable carrier on active component and the materia medica made the interior injection with dosage form or non-oral administration of oral administration according to conventional method; The medicine that can prepare the various dosage forms such as pill, powder, tablet, unguentum, powder, injection, water preparation, granule, injection of diseases such as becoming treatment coronary heart disease, angina pectoris, cerebral thrombosis or pulmonary infarction; The health food form can be liquid such as the picture refreshment drink, lactic acid beverage, flavoring agent is as additive, such as joining wheaten food class, fruit jelly, ice cream, curing food, Petaso, dessert etc.
The present invention has following advantage:
This invented technology is simple, and is easy to operate, and production cost is lower, and raw material sources are extensive, safely, have no side effect with a high credibility, and the active extract part of the Fructus Rubi leaf that obtains has significant antithrombotic and anticoagulant active, and medicinal effects is remarkable.
The specific embodiment
1. the preparation of Framberry leaf extract: the leaf with red raspberry (red raspberry is referred to as Fructus Rubi again in Chinese materia medica) is a raw material, adopt solvent extraction method, resin adsorption method, and, obtain the water or the pure water extract of Fructus Rubi leaf in conjunction with conventional drying method such as concentrate drying, lyophilization or spray dryinges.Described alcohol is the full monohydric alcohol CnH that closes with following general formula
2n+1OH, n are 1,2,3 or 4, or monohydric alcohol CnH is closed in insatiable hunger
2n-1OH, n are 2,3 or 4.
Red raspberry leaf water or pure water extract by macroporous adsorbent resin through the process that the mixed solvent elution of water-alcohol obtains the mixed solvent elution thing of different proportion water-alcohol are: pack into directly through the extract that solvent extraction method obtains: height ratio is 1: 3-1: 7 macroporous adsorptive resins, the water through containing different proportion-pure mixed solvent eluting and the eluate that obtains; Described alcohol is the full monohydric alcohol CnH that closes with following general formula
2n+1OH, n are 1,2,3 or 4, or monohydric alcohol CnH is closed in insatiable hunger
2n-1OH, n are 2,3 or 4; In eluting solvent, pure shared volumn concentration is between 5-95%.Resin comprises nonpolar macroporous adsorption resin, low pole macroporous adsorbent resin, polar macroporous adsorption resin, strong polar macroporous adsorption resin.Its eluate has confirmed to have anticoagulation and antithrombotic acitivity in zoopery.
1) preparation example
The macroporous adsorbent resin legal system is equipped with the red raspberry leaf effective-part: red raspberry leaf 1300 is restrained in the water that joins 8000 milliliters, through heating and refluxing extraction, remove by filter medicinal residues; Red raspberry leaf filtering residue joins heating and refluxing extraction in 8000 milliliters the water again, removes by filter medicinal residues; The red raspberry leaf is heating and refluxing extraction 2 times altogether, and each 2 hours, merge the water extract twice, 50-90 ℃ promptly obtains red raspberry leaf water extracting liquid through being evaporated to 1200 milliliters.Get 800 milliliters of above-mentioned red raspberry leaf water extracting liquids, extract is passed through nonpolar macroporous adsorption resin HPD-100, last bed volume is 1700 milliliters, and respectively with water, 30% ethanol, 70% ethanol and 95% ethanol gradient elution, elution volume is respectively 5100 milliliters.Eluate is through decompression and solvent recovery, and concentrate drying obtains the red raspberry leaf, water elution thing 166.23 grams, 30% ethanol elution thing, 49.7 grams, 70% ethanol elution thing, 14.84 grams, 95% ethanol elution thing, 1.28 grams.
2) adopt of the evaluation of LC-MS method to chemical constituent in the red raspberry leaf activity extract:
The anti-phase ODS post of immobile phase: Thermo (150 * 4.6mm, 5 μ m)
Mobile phase: 0-15min, 25: 75 (v: v) of methanol-water; 15-40min, 28: 72 (v: v) of methanol-water; 40-50min, 30: 70 (v: v) of methanol-water; 50-60min, 35: 65 (v: v) of methanol-water; 60-70min, 40: 60 (v: v) of methanol-water; 70-80min, 45: 55 (v: v) of methanol-water; 80-90min, 50: 50 (v: v) of methanol-water.
Wavelength: 256nm; Sample size: 5 μ l; Flow velocity: 0.5ml/min; Column temperature: 40 ℃; Ion source: ESI;
Through identifying that the main chromatographic peak of red raspberry leaf activity extract is: tilia glycosides (Tiliroside), 12,18-diene-28-glucoside-2 α, 3 β, acid in the 23-trihydroxy Usu, 12-alkene-28-glucoside-2 α, 3 β, acid in 19 α-trihydroxy-23-methyl Usu, 2,7-diethyl oxygen androstene, 12-alkene-28-glucoside-2 α, 3 β, acid in 19 α-trihydroxy Usu, 11,13-diene-28-glucoside-2 α, 3 α, 19 α-trihydroxy oleanolic acid.
More than 6 chemical compounds promptly may be for having anticoagulation and/or antithrombotic pharmaceutically active component.
2. red raspberry leaf anticoagulation and antithrombotic medicinal activity:
With red raspberry leaf 100g through the water reflux, extract,, extract is through nonpolar absorption macroporous resin, with the water of different proportion and the mixed solvent eluting of alcohol, obtain the water-pure mixed solvent eluting position of different proportion, its eluting position is carried out the experiment of the multiple calcium coagulation time test of blood plasma.
Red raspberry leaf 100 is restrained in the water that joins 1000 milliliters,, remove by filter medicinal residues through heating and refluxing extraction; Red raspberry leaf filtering residue joins heating and refluxing extraction in 1500 milliliters the water again, removes by filter medicinal residues; The red raspberry leaf is heating and refluxing extraction 2 times altogether, and each 1 hour, merge the water extract twice, 70 ℃ promptly obtain red raspberry leaf water extracting liquid through being evaporated to 500 milliliters.Get above-mentioned red raspberry leaf water extracting liquid and pass through nonpolar macroporous adsorption resin HPD-100 for 30 milliliters, last bed volume is 1700 milliliters, and respectively with water, 30% ethanol, 70% ethanol and 95% ethanol gradient elution, elution volume is 180 milliliters.Eluate is through decompression and solvent recovery, and concentrate drying obtains the red raspberry leaf, water elution thing 5.181 grams, 30% ethanol elution thing, 1.68 grams, 70% ethanol elution thing, 0.3731 gram, 95% ethanol elution thing, 0.0315 gram.
The water extract of the red raspberry leaf at the eluting position of red raspberry leaf 30～95% (V/V) concentration of alcohol is carried out following experiment;
2.1 external activity experiment: to the influence of blood plasma recalcification time
Get 30 of mices, be divided into three groups at random, 10 every group, normal saline group, administration group and Radix Salviae Miltiorrhizae group.Every eye socket is got blood 1ml, add to be placed with in the centrifuge tube of 3.8% sodium citrate 0.1ml, behind the mixing with 1000r/min
-1Centrifugal 10min.Get 10 in internal diameter 8mm test tube, add pooled plasma (blood is put into the centrifuge tube that contains 3.8% sodium citrate, and is centrifugal) and each 0.1ml of NS (normal saline) respectively, put into 37 ℃ of water-bath temperature immediately and bathe 1min, add 2.775g.L
-1CaCl
2Solution 0.1ml, mixing is still put into 37 ℃ of water-baths, begins to use manual time-keeping simultaneously, stops timing when detecting fiber protein yarn with bamboo let.Repeat 10 times, calculate 10 times meansigma methods, be matched group blood plasma recalcification time.Change NS into solution to be measured, obtain to be measured group of blood plasma recalcification time by same procedure.The results are shown in Table 1.
The eluting position of table 1 red raspberry leaf 30～95% determining alcohols is to the influence (n=10) of blood plasma recalcification time
Compare with matched group: * P＜0.05
Conclusion: the eluting position that shows red raspberry leaf 30～95% determining alcohols by the external activity test result can significant prolongation blood plasma recalcification time, and external blood coagulation resisting function is good, and may be used for the treatment of with the blood coagulation is the multiple disease of pathogenesis.
2.2 intracorporeal active experiment: ADP is brought out the influence of chmice acute pulmonary infarction
40 of male mice in kunming, body weight 18-22 gram is divided into four groups at random by the body weight equilibrium, 10 every group.ADP is China Medicine's product.The administration group is irritated the eluting position low dosage (26.8mg/kg) of stomach red raspberry leaf 30～95% determining alcohols and the eluting position high dose (53.6mg/kg) of red raspberry leaf 30～95% determining alcohols respectively, positive controls is irritated stomach Radix Salviae Miltiorrhizae crude extract (1.418g/kg that is equivalent to the crude drug amount), negative control group is irritated the normal saline (NS) of stomach same volume, every day 1 time, 4d continuously.Behind the last administration 1h, difference tail vein injection ADP normal saline solution 200mg/kg, mice breathes dyspnea with rapid and short breath immediately, and is stiff.To the time that mice recovers autonomic activities, the results are shown in Table 2 behind the record injection derivant.The high low dose group in eluting position of red raspberry leaf 30～95% determining alcohols and Radix Salviae Miltiorrhizae group all can significantly shorten the caused breathing dyspnea with rapid and short breath persistent period of the inductive platelet aggregation pulmonary infarction of ADP.
The influence (n=10) that the chmice acute pulmonary infarction forms is brought out to ADP in the eluting position of table 2 red raspberry leaf 30～95% determining alcohols
Compare with matched group: * P＜0.05
Conclusion: the eluting position low dosage of red raspberry leaf 30～95% determining alcohols uses and can make ADP bring out the mouse breathing dyspnea with rapid and short breath time obviously to shorten, and activity is better than positive control drug Radix Salviae Miltiorrhizae group, illustrates that it has certain therapeutical effect to acute pulmonary embolism.
2.3 interpretation
Red raspberry leaf of the present invention has anticoagulation and anti thrombotic action.The red raspberry leaf extract is at the external clotting time that prolongs.Adopt the thrombosed method of ADP, be similar to the pulmonary infarction of human body on the histology.Think at present ADP mainly be by with the P2Y1 receptors bind of platelet surface, promote Ca
2+In stream, cause platelet aggregation and form thrombosis.And experiment shows that the red raspberry leaf can prolong thrombus formation time, thereby plays antithrombotic pharmacological action.Thrombosis mainly comprises three phases: the fibrinous dissolving of (1) platelet adhesion and gathering (2) blood coagulation (3).The red raspberry leaf can suppress hematoblastic adhesion and gathering, and the energy blood viscosity lowering, and the latter also directly has influence on the thrombotic phase I.Hematoblastic minimizing will prolong the blood clotting time, thereby also have influence on the formation of thrombosis.
Claims (10)
1. Framberry leaf extract is characterized in that: it is the water of Fructus Rubi leaf or the extract of pure water.
2. according to the described Framberry leaf extract of claim 1, it is characterized in that: the water of described Fructus Rubi leaf or the concrete preparation process of the extract of pure water are, the Fructus Rubi leaf is mixed at the alcohol-water solution of 1%-70% with the 5-25 water or the volume content doubly of Fructus Rubi leaf volume, in 60-100 ℃ of lixiviate or reflux, extract, once or once, filter to collect filtrate, must have anticoagulation and antithrombotic acitivity plant extract.
3. according to the described Framberry leaf extract of claim 2, it is characterized in that: the time of each lixiviate or reflux, extract, is 1-24 hour; Described alcohol is the full monohydric alcohol CnH that closes with following general formula
2n+1OH, n are 1,2,3 or 4, or monohydric alcohol CnH is closed in insatiable hunger
2n-1OH, n are 2,3 or 4.
4. according to the described Framberry leaf extract of claim 2, it is characterized in that: the Fructus Rubi leaf water of collection or the filtrate of pure water extract are passed through macroporous adsorbent resin, adopt water-alcohol solvent to carry out gradient elution, the volume content of alcohol changes from 0-95% in the eluting solvent, collect the shared eluate of volumn concentration under the institute of the eluting solvent between 5%-95% eluting of alcohol in water-pure mixed solvent, eluate is reclaimed solvent through concentrating under reduced pressure, obtains having the plant extract of anticoagulation and antithrombotic acitivity.
5. according to the described Framberry leaf extract of claim 4, it is characterized in that: it is pressed powder that described eluate can adopt concentrate drying, lyophilization or spray drying process dry after concentrating under reduced pressure reclaims solvent, obtains having the plant extract of anticoagulation and antithrombotic acitivity.
6. the application of Framberry leaf extract in preparation anticoagulation and antithrombotic reagent, it is characterized in that: described Framberry leaf extract is used to prepare anticoagulation and/or antithrombotic reagent, anticoagulation and/or antithrombotic preventive health care's food or the food additive that prevents and/or treats to purpose; Described Framberry leaf extract is the water or the pure water extract of the described Fructus Rubi leaf of claim 1.
7. according to the application of the described Framberry leaf extract of claim 6 in preparation anticoagulation and antithrombotic reagent, it is characterized in that: the water of described Fructus Rubi leaf or the concrete preparation process of the extract of pure water are, the Fructus Rubi leaf is mixed at the alcohol-water solution of 1%-70% with the 5-25 water or the volume content doubly of Fructus Rubi leaf volume, in 60-100 ℃ of lixiviate or reflux, extract, once or once, filter to collect filtrate, must have anticoagulation and antithrombotic acitivity plant extract.
8. according to the application of the described Framberry leaf extract of claim 7 in preparation anticoagulation and antithrombotic reagent, it is characterized in that: the time of each lixiviate or reflux, extract, is 1-24 hour; Described alcohol is the full monohydric alcohol CnH that closes with following general formula
2n+1OH, n are 1,2,3 or 4, or monohydric alcohol CnH is closed in insatiable hunger
2n-1OH, n are 2,3 or 4.
9. according to the application of the described Framberry leaf extract of claim 7 in preparation anticoagulation and antithrombotic reagent, it is characterized in that: the Fructus Rubi leaf water of collection or the filtrate of pure water extract are passed through macroporous adsorbent resin, adopt water-alcohol solvent to carry out gradient elution, the volume content of alcohol changes from 0-95% in the eluting solvent, collect the shared eluate of volumn concentration under the institute of the eluting solvent between 5%-95% eluting of alcohol in water-pure mixed solvent, eluate is reclaimed solvent through concentrating under reduced pressure, eluate can be adopted concentrate drying after concentrating under reduced pressure reclaims solvent, lyophilization or spray drying process drying are pressed powder, obtain having the plant extract of anticoagulation and antithrombotic acitivity.
10. according to the application of the described Framberry leaf extract of claim 6 in preparation anticoagulation and antithrombotic reagent, it is characterized in that: described preparation prevents and/or treats to the anticoagulation of purpose and/or antithrombotic reagent are meant the described Framberry leaf extract of claim 1 is combined as excipient as acceptable carrier on active component and the materia medica, and its made pharmaceutical dosage form can be pill, powder, tablet, unguentum, powder, injection, water preparation, granule or injection.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102488641A (en) * | 2011-12-23 | 2012-06-13 | 郭保国 | Palmleaf raspberry leaf scrubbing bathing agent |
| CN105030810A (en) * | 2015-06-05 | 2015-11-11 | 湘潭大学 | Application of polysaccharide of rubus suavissimus S.Lee in inhibiting blood coagulation |
| CN113100366A (en) * | 2021-03-26 | 2021-07-13 | 中北大学 | Functional bubble water similar to beer and preparation method thereof |
| CN114306164A (en) * | 2022-01-11 | 2022-04-12 | 安徽农业大学 | Antibacterial acne-removing cream and preparation method thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1233282C (en) * | 2003-08-21 | 2005-12-28 | 孙相卿 | Bramble powder product preparing method |
| CN100374129C (en) * | 2004-12-14 | 2008-03-12 | 北京市农林科学院 | Framberry leaf extract effective part and its composition and uses |
-
2009
- 2009-03-04 CN CN 200910010568 patent/CN101822731B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102488641A (en) * | 2011-12-23 | 2012-06-13 | 郭保国 | Palmleaf raspberry leaf scrubbing bathing agent |
| CN102488641B (en) * | 2011-12-23 | 2013-04-17 | 郭保国 | Palmleaf raspberry leaf scrubbing bathing agent |
| CN105030810A (en) * | 2015-06-05 | 2015-11-11 | 湘潭大学 | Application of polysaccharide of rubus suavissimus S.Lee in inhibiting blood coagulation |
| CN113100366A (en) * | 2021-03-26 | 2021-07-13 | 中北大学 | Functional bubble water similar to beer and preparation method thereof |
| CN114306164A (en) * | 2022-01-11 | 2022-04-12 | 安徽农业大学 | Antibacterial acne-removing cream and preparation method thereof |
| CN114306164B (en) * | 2022-01-11 | 2023-09-22 | 安徽农业大学 | Antibacterial acne-removing cream and preparation method thereof |
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