CN101554399B - Psoralea corylifolial extract with immunologic enhancement and application thereof - Google Patents
Psoralea corylifolial extract with immunologic enhancement and application thereof Download PDFInfo
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- CN101554399B CN101554399B CN2009100987686A CN200910098768A CN101554399B CN 101554399 B CN101554399 B CN 101554399B CN 2009100987686 A CN2009100987686 A CN 2009100987686A CN 200910098768 A CN200910098768 A CN 200910098768A CN 101554399 B CN101554399 B CN 101554399B
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Abstract
The invention provides a psoralea corylifolial total glycoside extract which mainly contains 40 to 50 percent of psoralea corylifolial glycoside as well as 40 to 50 percent of isopsoralen glycoside. The psoralea corylifolial total glycoside extract and dressing or excipient allowed in pharmaceutics are prepared into any preparation formulation disclosed in pharmaceutics and can also be matched with other medicines or components to be prepared into a preparation together for use. Proved by pharmacological test, the psoralea corylifolial total glycoside and the psoralea corylifolial glycoside and the isopsoralen glycoside contained thereof can all resist the reduction of leucocyte caused by cyclophosphamide and have remarkable immunologic enhancement, and can be applied to the preparation ofthe medicines for increasing leucocyte.
Description
Technical field
The invention belongs to field of medicaments, particularly is a kind of Chinese herbal medicine extract, relates generally to the Fructus Psoraleae extract that contains malaytea scurfpea fruit total glycosides, and the application in the medicine of preparation leukocyte increasing.
Background technology
At present, in the world, Chinese herbal medicine all has certain market, along with people increasing and the aging of population to the health requirements level of understanding, sub-health stateization, people thirst for back to nature more, utilize the high Drug therapy of pure natural degree, prevent some chemical synthetic drugs cann't be solved problem, so the application of natural plant exceeds the background of its original traditional national culture.From natural drug, seek the little and inexpensive medicine of side effect and become the target that countries in the world pharmaceutical manufacturer is chased.The European Community has carried out unified legislation to medical herbs, state medical herbs status such as Canada and Australia have legalized, U.S. government has also drafted the plant amedica management method, the compound recipe mix preparation that begins to accept natural drug is as curative, and these provide good international environment for Chinese medicine enters international medical market as curative.On the other hand, along with the quickening of global economic integration progress, particularly China becomes a full member of WTO, and Chinese Medicine market incorporates the breadth and depth of international medical big market and will further aggravate.Face the enormous impact of Asian countries's traditional medicine product such as the keen competition of powerful transnational medical group and Japan, Korea S, India, Thailand and European countries' plant amedica such as Germany, France, numerous products that China's Chinese medicine produces are owing to still can not meet the standard of international medical market and requirement and being kept outside of the door.
The Chinese medicine Fructus Psoraleae is legumes psoraleae (Psoralea corylifolia L) fruit, and is warm in nature, acrid in the mouth, the effect of tool reinforcing the kidney and supporting YANG.Cure mainly the cold diarrhea of suffering from a deficiency of the kidney, frequent micturition, sexual impotence, chills and pain of the waist and kness, deficiency and coldness is breathed with cough, the external curing vitiligo.Modern study shows that Fructus Psoraleae has blood vessel dilating, increases myocardial contraction, and is antibiotic, the effect of estrogen sample, physiologically actives such as treatment vitiligo.Fructus Psoraleae contains the various active composition, and the glycoside that is representative with Fructus Psoraleae glycosides and different Fructus Psoraleae glycosides is one of its important active component, still is not that the medicine of main component emerges at present with the malaytea scurfpea fruit total glycosides.
Summary of the invention
The object of the present invention is to provide a kind of malaytea scurfpea fruit total glycosides extract, mainly containing percentage by weight is the Fructus Psoraleae glycosides of 40-50% and the different Fructus Psoraleae glycosides of 40-50%,
Fructus Psoraleae glycosides structural formula:
Different Fructus Psoraleae glycosides structural formula:
Another object of the present invention provides the application of described malaytea scurfpea fruit total glycosides extract in the medicine of preparation leukocyte increasing.
Malaytea scurfpea fruit total glycosides extract of the present invention, the dressing or the excipient that allow with preparation are prepared into said dosage form on any pharmaceutics, also can cooperate other drug or component to make preparation together and use.The dosage form of described medicine comprises liquid preparation, granule, tablet or soft gelatin capsule.The administering mode of described medicine comprises oral administration or drug administration by injection.
The present invention has obtained a kind of definite ingredients first, malaytea scurfpea fruit total glycosides extract evident in efficacy, the content of Fructus Psoraleae glycosides and different Fructus Psoraleae glycosides all can reach more than 40% in this malaytea scurfpea fruit total glycosides extract, pharmacological evaluation proves, this malaytea scurfpea fruit total glycosides with and the Fructus Psoraleae glycosides that contains and the different Fructus Psoraleae glycosides leucocytes reduction that all can resist caused by cyclophosphamide, have significant immunological enhancement.
Description of drawings
Fig. 1 malaytea scurfpea fruit total glycosides chromatogram.
Fig. 2 Fructus Psoraleae glycosides hydrogen spectrum.
Fig. 3 Fructus Psoraleae glycosides carbon spectrum.
Fig. 4 Fructus Psoraleae glycosides DEPT spectrum.
The different Fructus Psoraleae glycosides of Fig. 5 hydrogen spectrum.
The different Fructus Psoraleae glycosides of Fig. 6 carbon spectrum.
The different Fructus Psoraleae glycosides of Fig. 7 DEPT spectrum.
The specific embodiment
The present invention is further described in conjunction with the accompanying drawings and embodiments.
Embodiment one
Get Fructus Psoraleae 1kg, 50 ℃ of dryings 6 hours add 10 times of water gaging slight boiling condition heating and refluxing extraction 2 times, each 2 hours, extracting liquid filtering, concentrating under reduced pressure, precipitate with ethanol is regulated determining alcohol to 70%, gets supernatant, add ethyl acetate extraction 3 times after being concentrated into no alcohol, separate water layer, last macroporous resin, water wash to sugar-free (molish reaction), discard, continue with 70% ethanol water eluting.Collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets malaytea scurfpea fruit total glycosides.Through liquid phase analysis, contain two main components, content is respectively 40-50%, and referring to Fig. 1,1 is different Fructus Psoraleae glycosides among the figure, and 2 is the Fructus Psoraleae glycosides.Separation is obtained different Fructus Psoraleae glycosides and Fructus Psoraleae glycosides, do the hydrogen spectrum respectively, the carbon spectrum, the DEPT spectrum referring to Fig. 2-Fig. 7, can confirm that 1 is different Fructus Psoraleae glycosides, 2 is the Fructus Psoraleae glycosides.
The influence that embodiment two malaytea scurfpea fruit total glycosides reduce caused by cyclophosphamide normal mouse numeration of leukocyte
1. experiment purpose: investigate the influence of malaytea scurfpea fruit total glycosides to the caused by cyclophosphamide leucocytes reduction.
2. experiment material
2.1 medicine and reagent: Colla Corii Asini, Dong-E donkey-hide Gelatin Co., Ltd., Shandong Prov., lot number: 0311101; Cyclophosphamide for injection (CTX), Hengrui Medicine Co., Ltd., Jiangsu Prov., lot number: 7063021; The leukocyte diluent: glacial acetic acid 2ml, 1% Gentian Violet 1ml, distilled water adds to 100ml.
2.2 experiment equipment: cell counting count board, microscope, 1000 μ l, 100 μ l micropipettors.
2.3 laboratory animal: the ICR mice, male and female double, body weight 18-22g is provided by the Zhejiang Academy of Medical Sciences animal center.
3. method and step
3.1 get healthy 18-22g ICR mice, under this laboratory condition, raise after 2 days random packet:
Normal control group: isometric(al) normal saline
Model group: isometric(al) normal saline
Colla Corii Asini group: 1.5gKg
-1
Experimental group: low, high dose group (crude drug amount 2gKg
-1, 4gKg
-1);
Every group 10.Each organized the mice successive administration 8 days.
3.2 administration the 3rd day, every mouse peritoneal injection cyclophosphamide 100mg/kg (0.2ml/10g).
3.3 behind the 8th day (injection cyclophosphamide after the 5th day) administration 2h, the eye socket vein is got blood, draws 20 μ l blood, changes 380 μ l leukocyte diluents immediately over to, shakes gently 1～2 minute, fully counts behind the mixing.
4. experimental result
Experimental result shows, behind the injection cyclophosphamide the 5th day, the model group numeration of leukocyte was (3.33 ± 0.98) * 10
6/ ml is than normal group (7.24 ± 1.62) * 10
6/ ml obviously reduces, difference significance (P＜0.01).Low dose group and high dose group numeration of leukocyte are (4.05 ± 0.80) * 10
6/ ml, (4.29 ± 1.37) * 10
6/ ml obviously raises than model group, difference significance (P＜0.05, P＜0.05).Positive controls (Colla Corii Asini group) numeration of leukocyte is (4.05 ± 0.83) * 10
6/ ml obviously raises than model group, difference significance (P＜0.05).
The influence that table 1 malaytea scurfpea fruit total glycosides reduces caused by cyclophosphamide normal mouse numeration of leukocyte (x ± s, n=10)
Annotate: compare with normal group
*P＜0.05,
*P＜0.01;
Compare with model group
△P＜0.05,
△ △P＜0.01.
5. conclusion
The leucocytes reduction that malaytea scurfpea fruit total glycosides is low, high dose all can resist caused by cyclophosphamide.
The influence that embodiment three Fructus Psoraleae glycosides reduce caused by cyclophosphamide normal mouse numeration of leukocyte
1. experiment purpose: investigate of the influence of Fructus Psoraleae glycosides to the caused by cyclophosphamide leucocytes reduction.
2. experiment material
2.1 medicine and reagent
Colla Corii Asini, Dong-E donkey-hide Gelatin Co., Ltd., Shandong Prov., lot number: 0311101;
Cyclophosphamide for injection (CTX), Hengrui Medicine Co., Ltd., Jiangsu Prov., lot number: 07063021;
The leukocyte diluent: glacial acetic acid 2ml, 1% Gentian Violet 1ml, distilled water adds to 100ml.
2.2 experiment equipment: cell counting count board, microscope, 1000 μ l, 100 μ l micropipettors.
2.3 laboratory animal
The ICR mice, male and female double, body weight 18-22g is provided by the Zhejiang Academy of Medical Sciences animal center.
3. method and step
3.1 get healthy 18-22g ICR mice, under this laboratory condition, raise after 2 days random packet:
Normal control group: isometric(al) normal saline
Model group: isometric(al) normal saline
Colla Corii Asini group: 1.5gKg
-1
Experimental group: low, high dose group (0.1gKg
-1, 0.3gKg
-1);
Every group 10.Each organized the mice successive administration 8 days.
3.2 administration the 3rd day, every mouse peritoneal injection cyclophosphamide 100mg/kg (0.2ml/10g).
3.3 behind the 8th day (injection cyclophosphamide after the 5th day) administration 2h, the eye socket vein is got blood, draws 20 μ l blood, changes 380 μ l leukocyte diluents immediately over to, shakes gently 1～2 minute, fully counts behind the mixing.
4. experimental result
Experimental result shows, behind the injection cyclophosphamide the 5th day, the model group numeration of leukocyte was (3.33 ± 0.98) * 10
6/ ml is than normal group (7.24 ± 1.62) * 10
6/ ml obviously reduces, difference significance (P＜0.01).Low dose group and high dose group numeration of leukocyte are (3.82 ± 0.66) * 10
6/ ml, (3.99 ± 1.12) * 10
6/ ml obviously raises than model group, difference significance (P＜0.05, P＜0.05).Positive controls (Colla Corii Asini group) numeration of leukocyte is (4.05 ± 0.83) * 10
6/ ml obviously raises than model group, difference significance (P＜0.05).
The influence that table 2 Fructus Psoraleae glycosides reduces caused by cyclophosphamide normal mouse numeration of leukocyte (x ± s, n=10)
Annotate: compare with normal group
*P＜0.05,
*P＜0.01;
Compare with model group
△P＜0.05,
△ △P＜0.01.
5. conclusion
The leucocytes reduction that the Fructus Psoraleae glycosides is low, high dose all can resist caused by cyclophosphamide.
The influence that embodiment four different Fructus Psoraleae glycosides reduce caused by cyclophosphamide normal mouse numeration of leukocyte
1. experiment purpose: investigate of the influence of different Fructus Psoraleae glycosides to the caused by cyclophosphamide leucocytes reduction.
2. experiment material
2.1 medicine and reagent: Colla Corii Asini, Dong-E donkey-hide Gelatin Co., Ltd., Shandong Prov., lot number: 0311101; Cyclophosphamide for injection (CTX), Hengrui Medicine Co., Ltd., Jiangsu Prov., lot number: 07063021; The leukocyte diluent: glacial acetic acid 2ml, 1% Gentian Violet 1ml, distilled water adds to 100ml.
2.2 experiment equipment: cell counting count board, microscope, 1000 μ l, 100 μ l micropipettors.
2.3 laboratory animal
The ICR mice, male and female double, body weight 18-22g is provided by the Zhejiang Academy of Medical Sciences animal center.
3. method and step
3.1 get healthy 18-22g ICR mice, under this laboratory condition, raise after 2 days random packet:
Normal control group: isometric(al) normal saline
Model group: isometric(al) normal saline
Colla Corii Asini group: 1.5gKg
-1
Experimental group: low, high dose group (0.1gKg
-1, 0.3gKg
-1);
Every group 10.Each organized the mice successive administration 8 days.
3.2 administration the 3rd day, every mouse peritoneal injection cyclophosphamide 100mg/kg (0.2ml/10g).
3.3 behind the 8th day (injection cyclophosphamide after the 5th day) administration 2h, the eye socket vein is got blood, draws 20 μ l blood, changes 380 μ l leukocyte diluents immediately over to, shakes gently 1～2 minute, fully counts behind the mixing.
4. experimental result
Experimental result shows, behind the injection cyclophosphamide the 5th day, the model group numeration of leukocyte was (3.33 ± 0.98) * 10
6/ ml is than normal group (7.24 ± 1.62) * 10
6/ ml obviously reduces, difference significance (P＜0.01).Low dose group and high dose group numeration of leukocyte are (4.01 ± 1.02) * 10
6/ ml, (3.89 ± 0.65) * 10
6/ ml obviously raises than model group, difference significance (P＜0.05, P＜0.05).Positive controls (Colla Corii Asini group) numeration of leukocyte is (4.05 ± 0.83) * 10
6/ ml obviously raises than model group, difference significance (P＜0.05).
The influence that the different Fructus Psoraleae glycosides of table 3 reduces caused by cyclophosphamide normal mouse numeration of leukocyte (x ± s, n=10)
Annotate: compare with normal group
*P＜0.05,
*P＜0.01;
Compare with model group
△P＜0.05,
△ △P＜0.01.
5. conclusion
The leucocytes reduction that different Fructus Psoraleae glycosides is low, high dose all can resist caused by cyclophosphamide.
The preparation of embodiment five drop pill
Get malaytea scurfpea fruit total glycosides 0.5g and 10.5g Polyethylene Glycol-6000 mix homogeneously, heating and melting moves in the drop pill drip irrigation behind the change material, and in ℃ liquid paraffin of medicine liquid droplet to 6～8, oil removing makes 300 of drop pill.
The preparation of embodiment six lyophilized injectable powders
Get Lignum Dalbergiae Odoriferae oil 1.5g, join in the saturated hydroxypropyl of 13ml, stirring and dissolving filters, and the filtrate cold drying gets the clathrate powder of Lignum Dalbergiae Odoriferae oil and hydroxypropyl.Except that above-mentioned Lignum Dalbergiae Odoriferae oil closes the clathrate powder of hydroxypropyl, get malaytea scurfpea fruit total glycosides 0.5g, mannitol 5.5g, calcium disodium edetate 0.9g and distilled water 2ml again, behind the said components mixing, lyophilization, 350 of packing, promptly.
The preparation of embodiment seven tablets
Get malaytea scurfpea fruit total glycosides 100 gram,, add 3% polyvidone alcoholic solution system soft material, cross 18 mesh sieve system granules with the microcrystalline Cellulose mix homogeneously, 60 ℃ of dryings 1 hour, granulate, the adding Pulvis Talci is an amount of, mixing, tabletting, promptly.
Embodiment eight capsular preparations
Get Fructus Psoraleae glycosides and Oleum Arachidis hypogaeae semen in 2: 18 ratio (weight ratio) the water dissolving type rustless steel stirring filling, add an amount of gelatin and glycerol simultaneously and mix, emit reuse colloid mill defibrination, the mixed liquor that grinds stirs, and makes medicinal liquid; Under 40-50 ℃ of temperature, stir glycerol and distilled water miscible, again with the glycerin liquid temperature to 80-90 ℃, get gelatin and add wherein to mix and stir, become glue until off-bottom, glue was left standstill 4 hours, make offset plate with laminator and use for the pill operation; Above-mentioned medicinal liquid of making and offset plate are sent into the pellet press pill, did cylinder typing in 4 hours at 22-25 ℃ of temperature canyon then, again dry 16-20 hour of 25-30 ℃ of temperature canyon, the qualified soft gelatin capsule of pick, clean with 95% ethanol, dry 4 hours of 25-30 ℃ of canyon, promptly.
The preparation of embodiment nine injection
Getting different Fructus Psoraleae glycosides 100 grams (with embodiment seven) dissolves with 40 ℃ of waters for injection, add an amount of pharmaceutical carrier isotonic agent, the pH value of regulator solution is 7.0-8.0, adds to the full amount of water for injection, and removes thermal source with the ultrafilter ultrafiltration, after measuring pH value, use membrane filtration, after the packing, autoclaving, check, packing, promptly.
Claims (5)
1. malaytea scurfpea fruit total glycosides extract is characterized in that: extract contains Fructus Psoraleae glycosides that percentage by weight is 40-50% and the different Fructus Psoraleae glycosides of 40-50%, obtains by the following method:
Get Fructus Psoraleae 1kg, 50 ℃ of dryings 6 hours add 10 times of water gaging slight boiling condition heating and refluxing extraction 2 times, each 2 hours, extracting liquid filtering, concentrating under reduced pressure, precipitate with ethanol, regulate determining alcohol to 70%, get supernatant, add ethyl acetate extraction 3 times after being concentrated into no alcohol, separate water layer, last macroporous resin, water wash to sugar-free, discard, continue to collect eluent with 70% ethanol water eluting, being evaporated to does not have alcohol, lyophilization, promptly.
2. the application of a kind of malaytea scurfpea fruit total glycosides extract according to claim 1 in the medicine of preparation leukocyte increasing.
3. the application of a kind of malaytea scurfpea fruit total glycosides extract according to claim 1 is characterized in that: drug prepared also contains preparation allowable pharmaceutical excipients or dressing.
4. the application of a kind of malaytea scurfpea fruit total glycosides extract according to claim 3 is characterized in that: the dosage form of described medicine comprises liquid preparation, granule, tablet or soft gelatin capsule.
5. the application of a kind of malaytea scurfpea fruit total glycosides extract according to claim 4 is characterized in that: the administering mode of described medicine comprises oral administration or drug administration by injection.
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| CN103408558B (en) * | 2013-07-18 | 2015-08-19 | 中国人民解放军第四军医大学 | A kind of method extracting psoralen and isopsorapen based on cloud point extraction |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1446814A (en) * | 2003-03-07 | 2003-10-08 | 上海大学 | Method for preparing psoralen and isopsoralen |
| CN101406513A (en) * | 2008-12-03 | 2009-04-15 | 天津中医药大学 | Malaytea scurfpea fruit extract, pharmaceutical composition containing the same, and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1446814A (en) * | 2003-03-07 | 2003-10-08 | 上海大学 | Method for preparing psoralen and isopsoralen |
| CN101406513A (en) * | 2008-12-03 | 2009-04-15 | 天津中医药大学 | Malaytea scurfpea fruit extract, pharmaceutical composition containing the same, and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 杨帆 等.补骨脂提取工艺的比较研究.《中国中药杂志》.2005,第30卷(第16期),1-6. * |
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