CN1768823A - Application of anemarrhena total saponin in preparation of antifungal drug - Google Patents
Application of anemarrhena total saponin in preparation of antifungal drug Download PDFInfo
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- CN1768823A CN1768823A CN 200510017190 CN200510017190A CN1768823A CN 1768823 A CN1768823 A CN 1768823A CN 200510017190 CN200510017190 CN 200510017190 CN 200510017190 A CN200510017190 A CN 200510017190A CN 1768823 A CN1768823 A CN 1768823A
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- rhizoma anemarrhenae
- skin
- total saponins
- emulsifiable paste
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Abstract
The invention relates to the use of anemarrhena rhizome saponins (ZMZG) in preparing antifungal medicament, which comprises medicinal active constituent of ZMZG, conventional excipient, base material, and preservative agent. The medicament can be prepared into any of the clinically usable dosage forms.
Description
Technical field:
The invention belongs to the field of Chinese medicines, relate to the new pharmaceutical uses of a kind of Rhizoma Anemarrhenae total saponins (ZMZG) extract.
Background technology:
The Rhizoma Anemarrhenae is the dry rhizome of the liliaceous plant Rhizoma Anemarrhenae (Anemarrhena asphodeloides Bge.).Main product Hebei, Shanxi.With Yi County, Hebei institute product matter the best, claim " the Xilin Rhizoma Anemarrhenae ".The effect that has clearing away heat-fire, promotes the production of body fluid and moisturize cures mainly fever caused by exogenous pathogenic factors, the hyperpyrexia excessive thirst, and lung-heat type cough, interior-heat is quenched one's thirst, diseases such as dryness of the intestine constipation.Rhizoma Anemarrhenae total saponins is the active component that extracts from the Rhizoma Anemarrhenae, according to the literature, platelet aggregation is had the obvious suppression effect, and antiinflammatory action and antivirus action are also arranged.
Summary of the invention:
The objective of the invention is to propose a kind of antifungal medicament, and its practical dosage form is provided.Find out that from pharmacological evaluation Rhizoma Anemarrhenae total saponins has antifungic action preferably, can be used for fungus type treating for skin disease, and have no side effect, the development and use field of also having widened Rhizoma Anemarrhenae crude drug simultaneously.
Because the present invention discloses the drug effect of extraced total timosaponin aspect antifungal first, therefore Rhizoma Anemarrhenae total saponins is mixed with medicament separately or with other active components or adjuvant, is used for the antifungal effect.
Rhizoma Anemarrhenae total saponins of the present invention is a kind of active constituents of medicine, preparation process routinely, can Rhizoma Anemarrhenae total saponins be main active, add adjuvants such as conventional auxiliary type agent, solvent, solubilizing agent, substrate, antiseptic, make the dosage form that is suitable for using clinically.For example ointment, cream, tincture, paste, Emulsion, emulsion agent, gel etc.
The content of extraced total timosaponin of the present invention is in dosage form 1%---in 10% the scope, can administration every day 2 times or repeatedly, and preferably 10%, each administration 2 times, suitable route of administration is external, percutaneous drug delivery.
The Rhizoma Anemarrhenae total saponins extracting method takes Amberlyst process to extract, i.e. Rhizoma Anemarrhenae 1000g, decocting in water 3 times, each 2h filters, merge the water extract,, wash with water after colourless through macroporous resin adsorption, use ethanol elution, merge ethanol elution, decompression steams ethanol, gets pale yellow powder, vacuum drying promptly gets Rhizoma Anemarrhenae total saponins.
The specific embodiment:
Embodiment one: form (10g Rhizoma Anemarrhenae total saponins/100g ointment)
The ointment compound method:
Rhizoma Anemarrhenae total saponins 10.0g, stearic acid 9.6g, glyceryl monostearate 2.8g, liquid paraffin 4.8g, white vaseline 0.8g, lanoline 4.0g, triethanolamine 0.32g, ethyl hydroxybenzoate 0.1g distilled water are an amount of;
Method for making: stearic acid, glyceryl monostearate, liquid paraffin, white vaseline and lanoline are oil phase, put in the evaporating dish,, get ZMZG and be dissolved in the distilled water in being heated in the water-bath about 80 ℃, with triethanolamine, ethyl hydroxybenzoate is put in the beaker, about heating in water bath to 80 ℃.Waiting relaxing the bowels with purgatives of warm nature that water is slowly poured in the oil phase, and in water-bath, constantly be stirred to semi-solid, at room temperature stirring again closely to condensation, standby behind the autoclaving.
Embodiment two
Experiment 1, external antifungic action
Materials and methods
For the examination strain: trichophyton gypseum, microsporon gypseum, acrothesium floccosum (fungus section of dermatosis hospital of Chinese medical courses in general institute provides).
Culture medium: Sha Shi broth bouillon
Experimental technique: adopt liquid tube method,
With the fluid medium for preparing in gnotobasis condition branch is packed 12 test tubes into, every pipe 1ml, it is stock solution that Rhizoma Anemarrhenae total saponins is mixed with 50mg/ml with distilled water, get 1ml and add first in vitro, adopt the test tube doubling dilution to make 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.125mg/ml, 1.5625mg/ml, 0.7813mg/ml, 0.3906mg/ml, 0.1925mg/ml, 0.0977mg/ml, the medicinal liquid culture medium of 0.0488mg/ml series concentration.Each pastille in vitro adds the bacteria suspension (1 * 10 of 0.1ml
4-1 * 10
5Cfu/ml).The 11st pipe does not add bacterium liquid, and as negative control, the 12nd pipe adds bacterium liquid, as positive control.Put in 26 ℃ of calorstats and cultivate a week, observe its growing state every day, and keep a record.The highest drug dilution degree that fungus does not grow is the minimum inhibitory concentration (MIC) of this medicine.Take out medicinal liquid culture medium subcultivation that fungus do not grow to plate sabouraud's agar base, cultivate weeks observed result again in 26 ℃, the high dilution that fungus does not grow is the minimum bactericidal concentration (MBC) of this medicine.See Table 1
Table 1 Rhizoma Anemarrhenae total saponins is to three kinds of dermatophytosis experimental results
| Strain | MIC(mg/ml) | MBC(mg/ml) |
| Trichophyton gypseum microsporon gypseum acrothesium floccosum | 0.0977 0.7813 0.7813 | 0.0977 1.5625 1.5625 |
The result shows that Rhizoma Anemarrhenae total saponins is to trichophyton gypseum, and microsporon gypseum and acrothesium floccosum all have antibacterial and bactericidal action, and is especially comparatively remarkable to the effect of trichophyton gypseum.
Test the experiment of 2 therapeutic effect
Materials and methods
Medicine: Rhizoma Anemarrhenae total saponins emulsifiable paste; Emulsifiable paste matrix; Miconazole cream (Xian-Janssen Pharmaceutical Ltd., batch number 050406545)
Animal: 30 of healthy albino guinea-pigs (quality certification number is provided by Changchun High-technology Medical Animal Experiment Research Center: SCXK-(Ji) 2003-0004), male and female half and half, body weight 250g ± 50g.
Strain: trichophyton gypseum.
Method:
The bacteria suspension preparation
Hook up the ripe bacterium colony of the trichophyton gypseum of on the Sha Shi slant medium, growing with inoculation, be added in the physiological saline solution, after grinding homogenate, make (1～5) * 10 approximately
6The spore suspension of cfu/ml.
The preparation of Cavia porcellus infection model
Prepare smooth no hair-fields with depilatory at back, Cavia porcellus spinal column both sides, hair 4 * 5cm is shaved in every Cavia porcellus dorsal surface infected area, after 24 hours, artificial challenge's trichophyton gypseum according to a conventional method, after 3～5 days, local infection obviously forms, after mycology is checked the positive, determine to infect successfully.
Divide 3 groups at random with the Cavia porcellus of 30 infection, ZMZG emulsifiable paste group, emulsifiable paste matrix group, positive controls (miconazole cream), 10 every group.Every infection place dosage 0.2g 2 times/day, in continuous 2 weeks, carried out mycology microscopy and skin lesion in per 2 days and observes.The therapeutic evaluation standard is 2 grades: cure and be skin lesion continuous 2 feminine genders of fungus microscope examination that disappear; Invalidly do not disappear the fungus microscope examination positive for skin lesion.
The result
Rhizoma Anemarrhenae total saponins emulsifiable paste treatment Cavia porcellus local skin infects trichophyton gypseum, and the cure rate in two weeks is 85% (seeing Table 2); Miconazole cream treatment Cavia porcellus local skin infects trichophyton gypseum, and the cure rate in two weeks is 80% (seeing Table 2).The cure rate in the cure rate in two weeks of Rhizoma Anemarrhenae total saponins emulsifiable paste and two weeks of miconazole cream is relatively learned by statistics and is handled there was no significant difference (P＞0.05).Show that the ZMZG emulsifiable paste is close with daktarin on the dermatophytosis therapeutic effect.
Table 2 Rhizoma Anemarrhenae total saponins emulsifiable paste infects the therapeutic outcome of trichophyton gypseum to the Cavia porcellus local skin
| Group | The infected area number | One all cure rates | Two all cure rates |
| ZMZG emulsifiable paste group positive controls emulsifiable paste matrix group | 20 20 20 | 35％**# 30％** 0％ | 85％**# 80％** 10％ |
Annotate: emulsifiable paste matrix group and Rhizoma Anemarrhenae total saponins emulsifiable paste group, emulsifiable paste matrix group and positive controls compare: * * P＜0.05
Rhizoma Anemarrhenae total saponins emulsifiable paste group and positive controls compare: #P＞0.05
Test 3 skin irritation tests
Materials and methods
Medicine: Rhizoma Anemarrhenae total saponins emulsifiable paste, emulsifiable paste matrix.
Animal: 10 of healthy albino guinea-pigs (quality certification number is provided by Changchun High-technology Medical Animal Experiment Research Center: SCXK-(Ji) 2003-0004), male and female half and half, body weight 250g ± 50g.
Method: test preceding 24 hours with guinea pig back spinal column both sides depilations and not injured skins, the depilation area is 4cm * 5cm.The normal skin stimulation test is done in a depilation district, is divided into administration district and check plot.ZMZG ointment 0.2g is smeared in the administration district: the check plot is coated with the emulsifiable paste matrix with equivalent, covers the reuse immobilization with adhesive tape with layer of plastic film.Another piece depilation district makes the injured skin stimulation test, marks the scratch of " # " shape with syringe needle on depilation district skin before the administration, stabs epidermis, does not hinder corium, has slight oozing of blood to be degree.Medication is the same; After the administration 24 hours, remove medicine, write down the erythema of medicine-feeding part appearance in 1,24,48 and 72 hour and the situation of edema respectively, press table 3 scoring and estimate stimulus intensity.
Table 3 skin irritation reaction standards of grading
| Erythema | Integration | Edema | Integration |
| The visible reluctantly obviously visible moderate of no erythema is to serious erythema purple erythema and incrustation is arranged | 0 1 2 3 4 | The visible reluctantly obviously visible cutaneous protuberance profile of no edema is known that edema protuberance 1mm is above and is enlarged | 0 1 2 3 4 |
Table 4 Rhizoma Anemarrhenae total saponins ZMZG ointment is to skin irritant average response value
| Group matrix group normal skin matrix group injured skin ZMZG emulsifiable paste normal skin ZMZG emulsifiable paste injured skin | Number of animals | The integration summation | The average response value | ||||||
| 10 10 10 10 | 1 5 5 4 6 | 24 1 3 2 3 | 48 0 0 0 0 | 72 0 0 0 0 | 1 0.5 0.5 0.4 0.6 | 24 0.2 0.3 0.2 0.3 | 48 0 0 0 0 | 7 0 0 0 0 | |
The result
Visible ZMZG emulsifiable paste of table 4 and matrix group do not have significant difference to the average response value of normal skin and injured skin.The administration group is described to normal and the equal nonirritant of injured skin, matrix group is nonirritant also.
Test the experiment of 4 skin allergy
Materials and methods
Medicine: Rhizoma Anemarrhenae total saponins emulsifiable paste, 2, the 4-dinitrochlorobenzene, emulsifiable paste matrix.
Animal: 40 of healthy albino guinea-pigs, male and female half and half, body weight 250g ± 50g (carrying) by Changchun High-technology Medical Animal Experiment Research Center.
Method: divide 4 groups at random with animal, ZMZG emulsifiable paste, substrate matched group, blank group, positive controls (2,4-dinitrochlorobenzene), 10 every group.Test preceding 24 hours in the depilation of guinea pig back left side, the depilation area is 4cm * 5cm.Press the administration of skin semar technique.Each administration group of sensitization contact is coated with respectively with 0.2g (0.2ml) medicinal liquid in the unhairing district.The blank group is coated with the distilled water of equivalent, continues to remove after 6 hours to cover fixture.The 7th day and the 14th day, in kind repeat once, excited administration on the 28th day, with quadrat method several things that tried are applied to unhairing district, back part of animal skin right side respectively, 6 as a child flush away tried thing, observe at once, observed the skin allergy situation then once more in 24,48 and 72 hours, press the scoring of table 5 standard, and estimate the sensitization degree.
Table 5 skin allergy degree standards of grading
| Erythema | Integration | Edema | Integration |
| The obviously visible severe erythema purple erythema of the reluctantly visible moderate erythema of the slight erythema of no erythema also has incrustation | 0 1 2 3 4 | The obviously visible cutaneous protuberance 1mm profile of the reluctantly visible intermediate edema of no edema Mild edema is known that serious edema protuberance 1mm is above and is enlarged | 0 1 2 3 4 |
Table 6 Rhizoma Anemarrhenae total saponins (ZMZG) emulsifiable paste is to guinea pig skin influence hypersensitive
| Group ZMZG emulsifiable paste group substrate matched group positive controls blank group | Number of animals | The integration summation | The average response value | Quick rate (%) | ||||||
| 10 10 10 10 | 6 0 0 10 0 | 24 0 1 9 0 | 48 0 0 9 0 | 72(h) 0 0 0 0 | 6 0 0 1 0 | 24 0 0.1 0.9 0 | 48 0 0 0.9 0 | 72(h) 0 0 0 0 | 0 0 0 100 0 | |
The result
As shown in table 6, positive drug 2,4-dinitrochlorobenzene treated animal skin are subjected to the examination district to excite administration after 6 hours, have obviously slight erythema no edema, sensitization rate 100% to occur.Substrate control animals skin is subjected to examination district's animal show speckle slightly after exciting administration, recovers normal after 48 hours, thinks no sensitization phenomenon.ZMZG emulsifiable paste treated animal skin is subjected to the self-excitation administration of examination district erythema and edema not to occur after 6 to 72 hours, does not have difference with the blank group.This shows that the Rhizoma Anemarrhenae total saponins emulsifiable paste does not have sensitization.
Test 5 skin acute toxicity testings
Animal: 20 of healthy albino guinea-pigs (quality certification number is provided by Changchun High-technology Medical Animal Experiment Research Center: SCXK-(Ji) 2003-0004), male and female half and half, body weight 250g ± 50g.
Medicine: Rhizoma Anemarrhenae total saponins ointment
Method: animal is divided into 2 groups (being intact skin group and damaged skin group) at random, is tried preceding 24 hours guinea pig back both sides unhairing 4cm * 5cm, will be wherein 10 do damaged skin.Every depilation district is coated with the ZMZG ointment, and medicine was removed in administration in 24 hours, observed continuously 7 days.The result does not find any abnormal response, so think that this medicine does not have any acute toxic reaction to the Cavia porcellus external and occurs under this dosage.
Claims (2)
1, the application of Rhizoma Anemarrhenae total saponins in the antifungal drug of purchasing.
2, Rhizoma Anemarrhenae total saponins is a main active, adds conventional auxiliary type agent, solvent, solubilizing agent, substrate, antiseptic and makes ointment, cream, tincture, paste, Emulsion, emulsion agent, the gel that is suitable for using clinically.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510017190 CN1768823A (en) | 2005-10-17 | 2005-10-17 | Application of anemarrhena total saponin in preparation of antifungal drug |
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| CN 200510017190 CN1768823A (en) | 2005-10-17 | 2005-10-17 | Application of anemarrhena total saponin in preparation of antifungal drug |
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| CN1768823A true CN1768823A (en) | 2006-05-10 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110025717A (en) * | 2019-05-20 | 2019-07-19 | 上海中医药大学 | Timosaponin digests conversion product and inhibits the application in skin superficial fungi drug in preparation |
| CN115429736A (en) * | 2022-10-23 | 2022-12-06 | 爱生泽(上海)生物科技有限公司 | Preparation method of rhizoma anemarrhenae total saponin extract and application of rhizoma anemarrhenae total saponin extract in cosmetics |
-
2005
- 2005-10-17 CN CN 200510017190 patent/CN1768823A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110025717A (en) * | 2019-05-20 | 2019-07-19 | 上海中医药大学 | Timosaponin digests conversion product and inhibits the application in skin superficial fungi drug in preparation |
| CN110025717B (en) * | 2019-05-20 | 2022-02-08 | 上海中医药大学 | Application of timosaponin enzymatic hydrolysate in preparation of medicines for inhibiting skin superficial fungi |
| CN115429736A (en) * | 2022-10-23 | 2022-12-06 | 爱生泽(上海)生物科技有限公司 | Preparation method of rhizoma anemarrhenae total saponin extract and application of rhizoma anemarrhenae total saponin extract in cosmetics |
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