CN1923194B - Cortex fraxini coumarin composition - Google Patents
Cortex fraxini coumarin composition Download PDFInfo
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- CN1923194B CN1923194B CN2005100988089A CN200510098808A CN1923194B CN 1923194 B CN1923194 B CN 1923194B CN 2005100988089 A CN2005100988089 A CN 2005100988089A CN 200510098808 A CN200510098808 A CN 200510098808A CN 1923194 B CN1923194 B CN 1923194B
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Abstract
The invention relates to a method for preparing ash bark coumarin compound. Wherein, it boils and extracts with water, uses porous resin to separate and purify ash bark coumarin compound; and said compound comprises aesculin, aesculetin, fraxin, and fraxetin, while the total coumarin content is higher than 50%, and the contents of aesculin, aesculetin, fraxin, and fraxetin are higher than 40%, while their ratios are 4-6:1-3:2-4:1. The invention also provides its application in medicine production.
Description
Technical field
The present invention relates to a kind of preparation method and purposes of Chinese medicine Cortex Fraxini extract, more specifically say so and extract the total coumarins mixture that purification forms, its preparation method and pharmaceutical applications by Cortex Fraxini.
Background technology
Cortex Fraxini is Oleaceae plant fraxinus rhynchophylla Hance Fraxinus rhynchophylla Hance, Chinese ash Fraxinuschinensis Roxb., the dry bark of point leaf Chinese ash Fraxinus chinensis Roxb.var.acuminataLingelsh. or place post Chinese ash Fraxinus stylosa Lingelsh. or branch skin, contain with aseculin (aesculin), aesculetin (aesculetin), fraxin (fraxin), the Coumarins that 4 kinds of compositions of fraxetin (fraxetin) are representative becomes to grade, although existing at present bibliographical information Cortex Fraxini has anti-inflammatory and antalgic and the effect that reduces blood uric acid, clinically be used for hyperuricemia and gouty arthritis acute attack, the bibliographical information aseculin is also arranged, aesculetin, fraxin anti-inflammatory and antalgic and releasing uric acid pharmacology activity separately, but for the synergism of various Coumarins compositions in the Cortex Fraxini, not seeing has the research report.
Extraction separation purification for Coumarins composition in the Cortex Fraxini, existing bibliographical information all adopts decoction and alcohol sedimentation technique to obtain Cortex Fraxini extractum, it is dissolved in the hot water again, and cold preservation forms precipitation, leaching precipitates promptly, and what adopt in the patent application of Chengdu Dicon Institute Of Materia Medica also is the method.But the so-called total coumarins that the method makes is aseculin in fact just, and several Coumarins compositions of other that exists in the crude drug lose totally basically, and its pharmacologically active and clinical effectiveness significantly descend.
Chinese medicine is cured the disease, and emphasizes that each active component has now and studies have shown that in a large number effectively Chinese medicine curative effect after separation and purification reduces greatly originally for the Total tune effect of human body in the medical material.The inventor is by a large amount of chemistry and pharmacological screenings, proved that various Coumarins compositions are for the activity contribution and the synergism of treatment gout and hyperuricemia in the Cortex Fraxini, the cortex fraxini coumarin compositions that is obtained comprises a certain proportion of aseculin, aesculetin, fraxin, fraxetin, have antiinflammatory and uric acid resisting dual function, clinical usefulness is controlled gout and hyperuricemia determined curative effect.
The objective of the invention is by extraction and purification process research Coumarins composition in the Cortex Fraxini, obtain a kind of cortex fraxini coumarin compositions that to comprise aseculin, aesculetin, fraxin, 4 kinds of compositions of fraxetin be representative, wherein there are the ratio basically identical in the ratio of each constituent and its in crude drug, to improve the clinical effectiveness of its treatment gout and hyperuricemia.
Summary of the invention
Cortex fraxini coumarin compositions of the present invention obtains by the following method: get Cortex Fraxini, decoct with water and extract 3 times, add 10 times of amounts of water at every turn, extracted 1.5 hours, merge extractive liquid,, filter, filtrate is concentrated into about 1.1 (60 ℃ of heat are surveyed) of relative density, adds ethanol and makes and contain the alcohol amount and reach 80%, leave standstill, filter, decompression filtrate recycling ethanol adds water and makes dissolving in right amount to there not being the alcohol flavor, be added on the D101 type macroporous adsorptive resins of having handled well, with the water flushing, discard water lotion earlier, reuse 50% ethanol elution is to colourless, collect ethanol elution, reclaim ethanol to there not being the alcohol flavor, be concentrated into about 1.38 (60 ℃ of heat are surveyed) of relative density, vacuum drying, pulverize, cross 80 mesh sieves, mixing, promptly.
The total coumarins mixture that cortex fraxini coumarin compositions of the present invention is made up of a certain proportion of aseculin, aesculetin, fraxin, fraxetin etc., measure with ultraviolet absorption spectroscopy, contain total coumarins more than 50%, with high effective liquid chromatography for measuring, the total content of aseculin, aesculetin, fraxin, 4 kinds of compositions of fraxetin is more than 40%, 4 kinds of components in proportions are followed successively by 4-6: 1-3: 2-4: 1, and this ratio and its ratio basically identical in crude drug.
Discover that each composition has the obvious synergistic effect in the cortex fraxini coumarin compositions of the present invention, possesses various pharmacological activities such as gout arthritis and uric acid reducing simultaneously.Separation and purification goes out aseculin from this extract, and with purification before relatively, activity reduces greatly.
Cortex fraxini coumarin compositions of the present invention, can directly or adopt common pharmaceutical methods behind the compatibility, make the various dosage forms that are used for the treatment of gout and hyperuricemia, comprise capsule, soft capsule, tablet, granule, drop pill, injection and oral liquid formulation.
The present invention carries out the animal experiment study and the clinical trial of standard with the medicament of cortex fraxini coumarin preparation of compositions according to related request, proves that medicament has following obvious advantage and remarkable result.
1, toxicological experiment shows that the cortex fraxini coumarin compositions has avirulence
Cortex fraxini coumarin compositions of the present invention is extracted by the Chinese medicine Cortex Fraxini, and Cortex Fraxini derives from natural resources, is used as medicine through for a long time and uses proof, and Cortex Fraxini is taken for a long time and had no side effect.With the cortex fraxini coumarin compositions that Cortex Fraxini extracts, the active ingredient of enrichment medicine proves through toxicologic study, no overt toxicity.
Show through acute toxicity test in mice that (1) with the cortex fraxini coumarin compositions of extracting, the maximum tolerated dose of mouse stomach administration is 24g total coumarins/kg/24h, maximum tolerance multiple is 4000 times of 60kg body weight adult clinical oral administration day dosage.
Show through the rat long term toxicity test that (2) with the cortex fraxini coumarin compositions of extracting, the maximum tolerated dose of rat oral gavage administration is 18g total coumarins/kg/24h, maximum tolerance multiple is 3000 times of 60kg body weight adult clinical oral administration day dosage.The administration of rat oral gavage cortex fraxini coumarin compositions was observed 14 days, and rat does not have death, and only the visible animal activity of 15min to 18h reduces after administration, does not see other abnormal symptoms.After putting to death rat, all no abnormal discovery of perusal vitals (heart, liver, spleen, lung, kidney etc.).
(3) show through Beagle dog long term toxicity test, the cortex fraxini coumarin compositions is by 300,150 and three dosage of 75mg/kg, be equivalent to 50,25 and 12.5 times of its clinical oral administration dosage respectively, to Beagle dog oral administration 180 days and drug withdrawal 30 days, the result was as follows continuously: (1) does not have obviously influence to animal growth and behavioral activity etc.(2) routine urianlysis numerical value is all in normal range.(3) liver function test numerical value is all in range of normal value, but ALT, AST, T-Bil, ALP and/or the rising ALT effect of reduction arranged, and disappears after the drug withdrawal.(3) the kidney function test result value is all in normal range, but high, middle dosage group has rising creatinine and blood urea nitrogen effect, disappears after the drug withdrawal.(4) effect of the serum cholesterol of reduction is arranged.(5) Electrocardioscopy, result value all in normal range, but administration in the time of 45 days in dosage group decreased heart rate; Administration in the time of 180 days high dose group Q-T time interval significantly descend; Drug withdrawal 30 days, the R wave voltage of high, medium and low dosage group significantly, obviously and extremely significantly descends respectively.(6) living animal bone marrow examination, the result is all in normal range.But administration 180 days, visible low dose group basophilic erythroblast obviously raises.Obviously the raise staff cell of neutrophilic granulocyte of drug withdrawal 30 days, high dose group; In, low dose group obviously reduces lymphocyte.(7) system becomes celestial and organ index inspection Non Apparent Abnormality.(8) histopathologic examination, only find administration in the time of 180 days high dose group have 1 routine animal the slight focal inflammation pathological change of liver and 1 routine animal to occur the focal inflammation of slight kidney to occur and change, with matched group no significant difference (P＞0.05) relatively.All the other there is no unusually.
2, Expermental research on main pharmacodynamics proof cortex fraxini coumarin compositions has uric acid reducing and the effect of gout arthritis
(1) the cortex fraxini coumarin compositions is to the influence of mice Hypoxanthine type hyperuricemia formation
Use separately, only fraxin has certain intensity inhibitory action to suppress Hypoxanthine induced mice serum uric acid concentration rising effect (P＞0.05), and aseculin, aesculetin or fraxetin all do not have and obviously suppress H.And can obviously, significantly and extremely significantly suppress Hypoxanthine induced mice serum uric acid concentration rising (P＜0.05 respectively with cortex fraxini coumarin compositions 30,60 and the 120mg/kg that above-mentioned technology obtains, P＜0.01 and P＜0.001), its maximum uricopoiesis suppression ratio is 35.85%, is tangible dose-effect relationship (P＜0.05) in 30～120mg/kg dosage range.Its experiment the results are shown in Table 1.
The influence that table 1 cortex fraxini coumarin compositions forms mice Hypoxanthine type hyperuricemia
Annotate: 1. model control group and normal control group ^^^P＜0.001
2. administration group and model control group be * P＜0.05 * * P＜0.01 * * * P＜0.001 relatively
(2) rat urate excretion test
Table 2 shows, uses aseculin, aesculetin, fraxin or fraxetin 60～240mg/kg separately, does not all have obviously to promote rat urine urate excretion effect (P＞0.05).And cortex fraxini coumarin compositions 60 and 120mg/kg can significantly promote the drainage (P＜0.01) of rat uric acid through urine.
Table 2 cortex fraxini coumarin compositions is to the influence of rat urine urate excretion (x ± SD)
Annotate: administration group and distilled water matched group be * P＜0.05 * * P＜0.01 * * * P＜0.001 relatively
(3) uric acid sodium brings out rat gouty arthritis model test
Table 3 shows, uses aseculin, aesculetin, fraxin or fraxetin 60～240mg/kg separately, does not all have obviously rat gouty arthroncus effect (P＞0.05) due to the inhibition uric acid sodium.And above four components in certain proportion are followed successively by 4-6: 1-3: 2-4: 1 can significantly suppress rat gouty arthroncus (P＜0.01) due to the uric acid sodium for the cortex fraxini coumarin compositions of main constituent, remarkable inhibitory action (P＜0.01) was just arranged in 1 hour after the administration, keep 6h; 3h still has remarkable inhibitory action (P＜0.01) after the 30mg/kg administration.In 120～30mg/kg dosage range, be tangible dose-effect relationship (P＜0.05).
Table 3 cortex fraxini coumarin compositions is brought out the influence of rat gouty arthritis to uric acid sodium
Annotate: administration group and matched group be * P＜0.05 * * P＜0.01 * * * P＜0.001 relatively
(4) uric acid sodium brings out rabbit gouty arthritis model test
Table 4 cortex fraxini coumarin compositions is brought out the influence of rabbit gouty arthritis to uric acid sodium
Annotate: administration group and model control group be * P＜0.05 * * P＜0.01 relatively
Table 4 shows, use aseculin, aesculetin, fraxin or fraxetin separately, only 240mg/kg dosage group has rabbit gouty arthroncus effect (P＜0.05) due to the obvious inhibition uric acid sodium, and 120～60mg/kg dosage group does not all have obvious effect (P＞0.5).And above four components in certain proportion are followed successively by 4-6: 1-3: 2-4: 1 for occurring obviously suppressing rabbit gouty arthroncus effect (P＜0.05) due to the uric acid sodium after the cortex fraxini coumarin compositions 120mg/kg dosage group administration of main constituent in 1 hour, and effect is kept more than the 5h; 30mg/kg still has obvious inhibitory action (P＜0.05), is tangible dose-effect relationship (P＜0.05) in 120～30mg/kg dosage range.
Table 5 cortex fraxini coumarin compositions suppresses the effect that uric acid sodium stimulates rabbit synovial membrane liquid to generate
Annotate: 1. model control group and normal control group compare ^^^P＜0.001
2. administration group and model control group be * P＜0.05 * * P＜0.01 relatively
Table 5 shows, use aseculin, aesculetin, fraxin or fraxetin separately, only 240mg/kg dosage group has obvious inhibition uric acid sodium to stimulate the nucleus formation (P＜0.05) of rabbit synovial membrane liquid, and 120～60mg/kg dosage group does not all have obvious effect (P＞0.05).And above four components in certain proportion are followed successively by 4-6: 1-3: 2-4: 1 for the cortex fraxini coumarin compositions 120 of main constituent and the 60mg/kg remarkable and obvious generation (P＜0.01 and P＜0.05) that suppresses uric acid sodium stimulation rabbit synovial membrane liquid of energy respectively, is tangible dose-effect relationship in 30～120mg/kg scope.
The effect that the WBC number raises in the table 6 cortex fraxini coumarin compositions inhibition uric acid sodium stimulation rabbit synovial membrane liquid (x ± SD)
Annotate: 1. model control group and normal control group compare ^^^P＜0.001
2. each administration group and model control group compare * * P＜0.01 * * * P＜0.001
Table 6 shows, use aseculin, aesculetin, fraxin or fraxetin separately, only 240mg/kg dosage group has obvious inhibition uric acid sodium to stimulate the nucleus formation (P＜0.05) of rabbit synovial membrane liquid, and 120～60mg/kg dosage group does not all have obvious effect (P＞0.05).And above four components in certain proportion are followed successively by 4-6: 1-3: 2-4: 1 respectively can be extremely significantly and significantly suppress the migration (P＜0.001 and P＜0.05) that uric acid sodium stimulates WBC in the rabbit synovial membrane liquid for the cortex fraxini coumarin compositions 120 of main constituent and 60mg/kg group, is tangible dose-effect relationship in 30～120mg/kg scope.
Table 7 cortex fraxini coumarin compositions is to the influence of rabbit synovial membrane pathology due to the uric acid sodium
Annotate: compare * P＜0.05 * * P＜0.01 with model control group
Table 7 shows, use aseculin, aesculetin, fraxin or fraxetin separately, only 240mg/kg dosage group has rabbit knee synovial membrane congestion and edema due to the obvious inhibition uric acid sodium, crystallization calmness and synovial membrane fibrosis pathology variation effect (P＜0.05), and 120～60mg/kg dosage group does not all have obvious effect (P＞0.05).And above four components in certain proportion are followed successively by 4-6: 1-3: 2-4: 1 for cortex fraxini coumarin compositions 120～60mg/kg dosage group of main constituent has rabbit knee synovial membrane congestion and edema due to the remarkable inhibition uric acid sodium, crystallization calmness and synovial membrane fibrosis pathology variation effect (P＜0.01), is tangible dose-effect relationship in 30～120mg/kg scope.
Providing most preferred embodiment below further specifies the present invention but the present invention is not limited.
Embodiment 1
Get 100 kilograms of Cortex Fraxini branch skins, be cut into threadly, decoct with water and extract 3 times, add 1000 liters in water at every turn, decocted merge extractive liquid, 1.5 hours, filter, merge three times extracting solution, be concentrated into about 1.1 (60 ℃ of heat are surveyed) of relative density, add ethanol and make and contain the alcohol amount and reach 80%, leave standstill, filter, decompression filtrate recycling ethanol is to there not being the alcohol flavor, add water and make dissolving for 100 liters, be added on the D101 type macroporous adsorptive resins of having handled well, earlier with 300 liters of flushings of water, discard water lotion, 600 liters of eluting of reuse 50% ethanol are collected 50% ethanol elution, reclaim ethanol to there not being the alcohol flavor, be concentrated into about 1.38 (60 ℃ of heat are surveyed) of relative density, vacuum drying is pulverized, and crosses 80 mesh sieves, mixing, promptly get 7.12 kilograms of cortex fraxini coumarin compositionss, by raw medicinal herbs, yield is 7%.Each constituent content is in this cortex fraxini coumarin compositions: total coumarins 57.5% (ultraviolet spectrophotometry), aseculin 22%, aesculetin 8%, fraxin 13%, fraxetin 4%, its proportion of composing consistent with its proportion of composing in medical material (the HPLC chromatogram is seen accompanying drawing).
Get above gained cortex fraxini coumarin compositions 5217g (containing total coumarins 3000g), add an amount of dilution of starch and adjust total amount to 15000g, stirring and evenly mixing, cross 80 mesh sieves, add Pulvis Talci, mixing, incapsulate, make 50000, promptly get cortex fraxini coumarin composition capsule preparation.Prove that through clinical research the curative effect of treatment gout and hyperuricemia obviously is better than the aseculin preparation.
Description of drawings:
Fig. 1 cortex fraxini coumarin composition sample HPLC figure
The three-dimensional collection of illustrative plates of Fig. 2 cortex fraxini coumarin composition sample HPLC (1. aseculin; 2. fraxin; 3. aesculetin; 4. fraxetin).
Claims (1)
1. cortex fraxini coumarin compositions is characterized in that:
The total coumarins mixture that a, this cortex fraxini coumarin compositions are made up of a certain proportion of aseculin (aesculin), aesculetin (aesculetin), fraxin (fraxin), fraxetin (fraxetin) etc., measure with ultraviolet absorption spectroscopy, contain total coumarins more than 50%;
Mainly contain aseculin (aesculin), aesculetin (aesculetin), fraxin (fraxin), 4 kinds of compositions of fraxetin (fraxetin) in b, this cortex fraxini coumarin compositions, with high effective liquid chromatography for measuring, the total content of 4 kinds of compositions is more than 40%, and 4 kinds of components in proportions are followed successively by 4-6: 1-3: 2-4: 1;
Described cortex fraxini coumarin preparation of compositions method comprises the steps: to get Cortex Fraxini, decocts with water to extract 3 times, adds 10 times of amounts of water at every turn, extracted 1.5 hours, merge extractive liquid, filters, filtrate is concentrated into 60 ℃ of heat and surveys relative densities 1.1, adds ethanol and makes and contain the alcohol amount and reach 80%, leaves standstill, filter, decompression filtrate recycling ethanol adds water and makes dissolving in right amount to there not being the alcohol flavor, be added on the D101 type macroporous adsorptive resins of having handled well, with the water flushing, discard water lotion earlier, reuse 50% ethanol elution is to colourless, collect ethanol elution, reclaim ethanol to there not being the alcohol flavor, be concentrated into 60 ℃ of heat and survey relative density 1.38, vacuum drying, pulverize, cross 80 mesh sieves, mixing, promptly.
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| CN102210787A (en) * | 2011-06-10 | 2011-10-12 | 中国药科大学 | Application of philippine violet herb total phenol extract to treatment of hyperuricemia |
| CN102293811B (en) * | 2011-08-01 | 2013-06-05 | 山东省中医药研究院 | Extraction method of total coumarin and application of extracted total coumarin |
| CN102631419B (en) * | 2012-04-28 | 2014-04-02 | 重庆市中药研究院 | Medicine composition for treating gout, as well as preparation method and application of medicine composition |
| CN102697827B (en) * | 2012-07-02 | 2014-07-02 | 新疆农业大学 | Coumarin from Cichorium plants used for preparing drug with liver-protecting effect, and extraction and purification method of coumarin from Cichorium plants |
| CN104311616B (en) * | 2014-09-30 | 2016-08-24 | 陕西省西安植物园 | A kind of extraction high purity cortex fraxini and method of fraxin from Cortex Fraxini |
| CN107501220A (en) * | 2017-08-23 | 2017-12-22 | 佛山市三水区嘉华化学研究院（普通合伙） | A kind of aesculetin and preparation method and application with antioxidant and anti-aging activity |
| CN114767737B (en) * | 2022-06-21 | 2022-11-29 | 上海中医药大学 | Cortex fraxini extract, preparation method and application thereof |
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| CN1398861A (en) * | 2002-08-29 | 2003-02-26 | 成都迪康药物研究所 | Prepn and application in preparing medicine of Fraxinus general coumarin |
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