CN1233330C - Extraction of Malassezia for treating chlosma, said prepn. and its prepn. method and application thereof - Google Patents
Extraction of Malassezia for treating chlosma, said prepn. and its prepn. method and application thereof Download PDFInfo
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- CN1233330C CN1233330C CNB031380433A CN03138043A CN1233330C CN 1233330 C CN1233330 C CN 1233330C CN B031380433 A CNB031380433 A CN B031380433A CN 03138043 A CN03138043 A CN 03138043A CN 1233330 C CN1233330 C CN 1233330C
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Abstract
The present invention relates to Malassezia bacterin for treating chloasma, a preparation, a preparation method and application thereof. The Malassezia preparation which comprises Malassezia bacteria and auxiliary materials can be prepared by mixing Malassezia bacteria and auxiliary materials. The preparation method of the Malassezia bacterin comprises: collecting and culturing Malassezia bacteria and preparing filter liquid. The preparation can be any external preparation for skin, especially solution, paste and emulsion. The present invention also relates to application of Malassezia bacterin for treating chloasma in the aspects of preparing medicine for treating chloasma disease symptoms and products for beautifying faces and protecting skin. The present invention has the advantages of high obvious effect rate, quick curative effect, short course of treatment, no toxic and side effect and low relapse rate.
Description
Technical field
The present invention relates to a kind of external preparation for the treatment of chloasma and its production and application, relate in particular to a kind of chlosma external used medicine for the treatment of chloasma and its production and application, be particularly related to a kind of chlosma (malassezia furfur, latin name Malassezia) rhzomorph for the treatment of chloasma, its preparation and its production and application.
Background technology
Chloasma is to betide the common skin diseases that human body face skin hinders beauty treatment, and its principal character is that skin of face pigment is in various degree deepened.
The pathogeny of chloasma and reason are not fully aware of, are common in gestation, oral contraceptive or agnogenio, still do not have satisfied Therapeutic Method at present, and its main pathology is characterized as in the epidermis melanocyte and produces excessive melanin granule and be deposited in the skin.Except that systemic administration, the most frequently used external anti-freckle agent has 2% Hydrochinone Cream at present, 2% kojic acid frost, 20% king's diacid frost, 0.01% retinoic acid frost, 5% ammoniated mercury ointment, 3% hydrogenperoxide steam generator, superoxide dismutase (SCD) frost etc., all there is following problem in above-mentioned Therapeutic Method: 1, obvious effective rate is low, curative effect is slow, and the course of treatment is long; 2, toxicity often appears, as flush, scorching hot, (except the common skin allergies) such as xerosis cutis and desquamations (document is seen " sick research and the prospect of learning of beauty treatment skin ", Lai Yongxian. raise uncommon Hui Zhonghua medical cosmetology magazine, 164 pages-166 pages of 2002 the 3rd phases); 3, relapse rate height, the above-mentioned treatment responder of clinical observation needs long-term prescription, often promptly recurs after drug withdrawal etc.
In treatment chloasma all medicine for external use of this dermatosis and the cosmetics, still there is not the chlosma of use rhzomorph at present as raw material.
The 5th edition 78-79 page or leaf of chlosma (malassezia furfur Malassezia) " skin cypridology ". the chief editor. open the high medical college teaching material in the People's Health Publisher .2001 JIUYUE whole nation of learning military affairs.
Summary of the invention
The purpose of this invention is to provide a kind of external preparation for the treatment of chloasma and its production and application, a kind of chlosma external used medicine for the treatment of chloasma and its production and application especially is provided, a kind of chlosma rhzomorph for the treatment of chloasma, its preparation and its production and application are provided especially, overcome that present treatment chloasma medicine obvious effective rate is low, curative effect is slow, the course of treatment is long, toxicity often appears, the shortcoming that relapse rate is high.
The present invention is based on the following mechanism of curing the disease:
According to clinical observation and classical documents (clinical dermatology. second edition. chief editor Zhao distinguishes. Jiangsu science tech publishing house in January, 1989) record, can cause tinea versicolor though pathogenic chlosma is clinical, but this chlosma also is the normal human bacterial parasite, only under given conditions, just can cause a disease as skin greasiness heavy wool, and a little less than the pathogenicity, only invade epidermis and do not invade corium and subcutaneous tissue.
The main pathological characters of chloasma is that melanocyte produces too much melanin granule in the skin, and the clinicopathological characteristics of tinea versicolor is to occur the hypopigmentation of skin speckle in the tinea versicolor morbidity after 1-2 month, and this depigmentation macule still can continue the several months after tinea versicolor is recovered even more than half a year, confirm that chlosma can produce the chlosma element in the growth and breeding process, this rhzomorph can suppress human melanocyte and produce melanin granule.The tinea versicolor clinical manifestation of causing a disease due to the back is slight, no inflammatory reaction and have only the change of pigment.
In view of the above, we carry out a large amount of artificial cultures after the tinea versicolor patient collects chlosma on one's body, will collect bacterium colony and make solution, and the preparation external preparation is used for the treatment of chloasma and obtains good result behind the autoclaving, and can not cause untoward reaction to human body skin.
The objective of the invention is to be achieved through the following technical solutions:
A kind of chlosma preparation for the treatment of chloasma comprises chlosma rhzomorph and adjuvant;
Described adjuvant can contain any nontoxic skin with conventional medicine or cosmetic activity composition, as materials such as opacifier, penetration enhancers, humidizer, surfactant, lubricant, coloring agent, toner, astringents;
Described adjuvant can be any skin usual excipients and additive, comprise antiseptic, thickening agent and spice, and conventional antioxidant, as BHA (BHA), ascorbyl palmitate, propyl gallate, citric acid, two BHAs (BHT), ethoxy quinoline etc., can also contain thickening agent well known in the art, cosolvent, wetting agent, emulsifying agent and viscosity stabiliser etc.;
Described preparation can be any preparation for external application to skin, especially is solution, unguentum, Emulsion;
The preparation method of the chlosma preparation of described treatment chloasma comprises chlosma rhzomorph and adjuvant are mixed;
The preparation of described chlosma rhzomorph may further comprise the steps at least: the collection cultivation of chlosma and the preparation of bacterium liquid; Described collection cultivation comprises by the culture medium of following parts by weight of component makes: get Semen Glycines powder and add the mid-refrigerator overnight taking-up of distilled water; Filter, add peptone 1 in 100 parts in the filtrate: agar 1.8: the ratio of glucose 4 adds mentioned component; Culture medium is made in the heating and melting sterilization; Inoculation was cultivated 5-7 days for 37 ℃;
The preparation of described bacterium liquid comprises that the chlosma tongue of getting cultivation is dissolved in the normal saline, makes bacteria suspension; And/or, the lawn of cultivating is inoculated in the fluid medium, put in 37 ℃ of shaking table culture medium, cultivated 5-7 days, make bacteria suspension; Above-mentioned bacteria suspension filtration sterilization promptly.
The preparation of described chlosma rhzomorph is further comprising the steps of: described collection cultivation comprises by the culture medium of following parts by weight of component makes, and gets 1000 parts of mid-refrigerators of distilled water of 40 parts of addings of Semen Glycines powder and crosses liquid taking-up next day; With filter paper or 8 layers of filtered through gauze, add mentioned component in 100 parts of ratios that add 1 part of peptone, 1.8 parts in agar, 4 parts of glucoses in the filtrate; Heating and melting, packing, 10 pounds of sterilizations in 10 minutes, it is standby to put the inclined-plane; To prepare the liquid branch and pack in the conical flask, it is standby to sterilize, and makes solid medium;
Culture medium is preferably husky fort culture medium, and PH is 6.5.The bacterial classification inoculation amount is for exceeding to inoculate successfully in right amount; The sterilization pressure be 10 pounds 10 minutes.
Describedly draw materials and cultivate to tinea versicolor patient skin lesion district is scraped and get squama, be inoculated in the surface of above-mentioned solid medium, cultivated 5-7 days for 37 ℃;
Behind the described bacteria suspension sterilization, filter standby; Describedly put sterilization behind 37 ℃ of shaking table culture medium culturings, cross the elimination sediment, standby.
The every ml of described bacteria suspension contains the about 30-90 of chlosma hundred million, is preferably 50-70 hundred million, especially is about 60 hundred million, and promptly the 100g pharmaceutical adjunct adds bacteria suspension 5ml.
A kind of application of chlosma preparation aspect preparation treatment chloasma medicine for the treatment of chloasma, described preparation contains chlosma rhzomorph and skin pharmaceutic adjuvant.
A kind of application of chlosma preparation aspect preparation treatment chloasma cosmetics and skincare product for the treatment of chloasma, described preparation contains described chlosma rhzomorph and skin protection adjuvant commonly used.
Preparation of the present invention is used for the frequency of utilization of skin should be decided according to patient and user individuality.It is fixed that concrete using method should be come according to age, body weight and the skin of individual patient.
The present invention has the following advantages:
1, preparation of the present invention can be made into various preparations, and is easy to use.；
2, obvious effective rate height of the present invention, curative effect is fast, short treating period.
3, except that indivedual normal skin anaphylaxiss, no toxicity, scorching hot as flush, phenomenons such as xerosis cutis and desquamation occur.
4, relapse rate is low after the drug withdrawal.
For proving effect of the present invention, Drug therapy chloasma 50 routine effect observations of the present invention are as described below.
The clinical diagnosis of chloasma and curative effect determinate standard
One, clinical diagnosis standard:
(1) facial filbert to dark brown, well-defined patch, symmetry distributes usually, NIP performance and squama
(2) there is not obvious subjective symptoms.
(3) mainly occur in adolescence after, women's pilosity.
(4) state of an illness has certain seasonality, and Xia Chongdong is light.
(5) do not have obvious endocrinopathy, and get rid of the pigmentation that other disease causes.
(6) average optical density value of pigmented area is greater than self more than 20% of facial average optical density value.
Two, curative effect determinate standard:
(1) the basic healing: (1) naked eyes are looked the mottle area and are disappeared greater than 90%, and color disappears substantially.
(2) point system is calculated treatment back decline index 〉=0.8.(3) the pigmented area skin image is measured the ID of efficacy evaluation unit 〉=55.
(2) produce effects: (1) naked eyes are looked the mottle area and are disappeared greater than 60%, and color is obviously thin out.
(2) point system is calculated treatment back decline index 〉=0.5.(3) the pigmented area skin image is measured the ID of efficacy evaluation unit 〉=15.
(3) take a turn for the better: (1) naked eyes are looked the mottle area and are disappeared greater than 30%, and color is thin out.
(2) point system is calculated treatment back decline index 〉=0.3.(3) the pigmented area skin image is measured the ID of efficacy evaluation unit 〉=5.
(4) invalid: (1) naked eyes are looked the mottle area and are disappeared less than 30%, and change color is not obvious.
(2) point system is calculated treatment back decline index 〉=0.(3) the pigmented area skin image is measured the ID of efficacy evaluation unit 〉=1.
Illustrate: the pigmented area skin image is measured the computational methods of efficacy evaluation list ID value: 1. optical density (OPT) is calculated: certain some gray scale (color) is A in hypothetical target (skin) image, incident illumination (specific light) demarcate be B, the optical density value of this point be OPT=Lng (B/A) so=LngB-LngA is because B 〉=A OPT 〉=0.2. integral optical density (OPTDM) calculates: supposition area (AREA) is M * N, and integral optical density is the summation of each point optical density in the area; Average optical (OPTDI) is the optical density meansigma methods in the integral optical density, that is:
3. ID value: represent skin image to measure efficacy evaluation unit, according to before the pigmentation treatment of patient with chloasma skin lesion zone and the change of the integral optical density value after the treatment, and the relative value who calculates.
Three, methods of marking and standard:
| Skin lesion distributes | The area size | The pigment degree | |
| 0 1 2 3 | Facial general of no face two sided bilateral | 0 ＜2cm 2 2～4cm 2 ＞4cm 2 | There is not the heavy dark brown of obvious brown in the pastel brown |
Four, point system decline index calculation method: total mark before decline index=(total mark one treatment back total mark before the treatment)/treatment
Drug therapy chloasma 50 routine effect observations of the present invention:
1, physical data
50 routine patients are my out-patient of institute and the worker of the court, women's 48 examples wherein, male's two examples, 22～45 years old age, average 32.7 years old, married 44 people, unmarried 6 people.The course of disease 3 months～10 years, average 3 years, all patients all diagnosed clearly.Diagnostic criteria is as the criterion with " clinical diagnosis of chloasma and the clinical development standard (draft) " that Chinese combination of Chinese and Western medicine department of dermatologry pigment disease group committee of association (in August, 1998 Harbin) formulates, wherein 5 people are morbidity before and after the gestation, menstruation deviant 11 people, surplus no obvious inducement.
2, Therapeutic Method
Conventional cleaning facial outer cream of the present invention every day each 1 time sooner or later of being coated with, be coated with back forlement 1 ~ 3 minute, other all oral and external medicine for removing speckle of stopping using during the medication, the skin nursing speckle dispelling treatment of stopping using.1 record effect of further consultation in per 15～30 days.Do not see any untoward reaction and allergic phenomena in the therapeutic process.
3, clinical development
According to curative effect determinate standard in " standard ", former 2 is main, and be almost recovered: skin lesion disappears greater than 90%, and color disappears substantially; Produce effects: skin lesion disappears greater than 60%, and color is obviously thin out; Effectively: skin lesion disappears greater than 30%, and color is thin out; Invalid: skin lesion disappears less than 30%, and change color is not obvious.
4, result
In 50 examples, 9 examples that are almost recovered (18%), produce effects 15 people (30%), effective 19 people (38%), invalid 7 people (14%), total effective rate 86%, respond well person is many to take effect in medication in 10 ~ 15 days, 2～3 months produce effects or recovery from illness.Medicine of the present invention (A group) is compared with traditional medicine for removing speckle compound recipe kojic acid frost (B group), and effect is better than the latter, compound recipe kojic acid frost case 23 examples.(quote Chinese medical beauty treatment magazine and rolled up 277 pages of the 5th phases " clinical observation of compound recipe kojic acid frost treatment pigmented dermatosis " in 2000 the 6th).
Two groups of patient effects relatively
| Group | Total routine number | Recovery from illness | Produce effects | Effectively | Invalid | Total effective rate (%) |
| A B adds up to | 50 23 73 | 9 1 10 | 15 3 18 | 19 9 28 | 7 9 16 | 86 61 |
| Statistical procedures P＜0.01 | ||||||
Annotate: the A group: speckle enlightening frost group B group: compound recipe kojic acid frost group
Concrete case
Case 1: Liu so-and-so. women .38 year, chloasma moderate. medical history 10 years.Use chlosma preparation affected part of exterior coating every day 2 times, during the treatment without other any oral and external application medicines.Produce effects after January was fully recovered after February.
Case 2: Lee woman 30 years old, chloasma severe, medical history 3 years, stop using other all in medicine for external use and cosmetics, behind the external chlosma preparation February surplus being clearly better.
Case 3: Deng woman 34 years old, the moderate chloasma, medical history 5 years, list is fully recovered after February with the chlosma preparation, nothing recurrence in 2 years so far.
Case 4: 37 years old chloasma moderate of so-and-so woman of Liu, recovery from illness adhered to using the chlosma preparation 2 years after 3 months.Skin lesion is not violated all the time, any side reaction do not occur and cut.
Case 5: 40 years old chloasma severe of so-and-so woman of Lee, medical history 8 years, external chlosma preparation is existing the effect after January, recovery from illness after 3 months.
The specific embodiment
Embodiment 1
The chlosma solution
Below the preparation method step of the chlosma rhzomorph of treatment chloasma:
1, the preparation of the collection of chlosma cultivation and filtrate
The preparation of culture medium: desirable Semen Glycines powder 40g adds the mid-refrigerator of 1000ml distilled water and crosses liquid taking-up next day; With 8 layers of filtered through gauze, get filtrate, the ratio that adds peptone 1g, agar 1.8g, glucose 4g in 100ml adds mentioned component; Heating and melting, packing, 10 pounds of sterilizations in 10 minutes, it is standby to put the inclined-plane; The preparation fluid medium does not add agar, 100ml is prepared the liquid branch pack in the conical flask of 250ml, and it is standby to sterilize.Every l00ml fluid medium adds 3.6g agar and makes solid medium, and it is standby to sterilize, and wherein, culture medium PH is 6.5.
2, draw materials and cultivate: tinea versicolor patient skin lesion district is carried out disinfection with 75% ethanol, gently scrape with aseptic nipper and to get squama, an amount of squama is inoculated in the surface of above-mentioned solid medium, cultivated 6 days for 37 ℃, when the inclined-plane in solid culture medium grows up to typical bacterium colony, stop cultivating.
3, the preparation of chlosma filtrate: the filtrate preparation method is that following two kinds (1) get solid culture inclined-plane lawn, is dissolved in the normal saline, makes bacteria suspension, behind the sterilization, filters and uses.(2) lawn of getting on 2 ~ 3 inoculating loop solid slant culture bases is inoculated in the fluid medium, puts in 37 ℃ of shaking table culture medium, cultivates autoclave sterilization sterilization after 6 days, crosses the elimination sediment, uses.
A kind of chlosma preparation for the treatment of chloasma is made up of described chlosma rhzomorph; Be that the filtrate of above-mentioned cultivation mix homogeneously behind autoclaving (10 pounds 10 minutes) is got final product.
Wherein, the every ml of bacteria suspension contains about 60 hundred million of chlosma.
Embodiment 2
The chlosma ointment
The preparation method of the chlosma rhzomorph of treatment chloasma is with embodiment 1, and difference is to get filtrate with the immersion of filter paper filtering bean, and the ratio that adds peptone 2g, agar 3.6g, glucose 8g in 100ml adds mentioned component, makes solid chlosma culture medium; Cultivated 5 days for 37 ℃, culture medium PH is 6.5.Described above-mentioned cultivation lawn is made filtrate mix homogeneously after 10 pounds of sterilizations in 10 minutes of high pressure and is got chlosma rhzomorph liquid.
A kind of chlosma ointment for the treatment of chloasma, form by described chlosma rhzomorph liquid and excipient, described excipient is for adding octadecanol 4 grams, stearic acid 4 grams, glyceryl monostearate 3.5 grams, vaseline 10 grams, glycerol 10 grams, soil temperature-802 gram adds surplus chlosma rhzomorph liquid and is mixed with ointment 100 grams.
Wherein, the every ml of bacteria suspension contains about 30 hundred million of chlosma, and promptly the 100g cream base adds bacteria suspension 2.5ml.
Embodiment 3
The chlosma emulsion agent
The preparation method of the chlosma rhzomorph of treatment chloasma is with embodiment 1, and difference is to get filtrate with the immersion of filter paper filtering bean, and the ratio that adds peptone 2g, agar 3g, glucose 6g in 100ml adds mentioned component, makes solid chlosma culture medium; Cultivated 7 days for 37 ℃, culture medium PH is 6.5.Described above-mentioned cultivation lawn is made filtrate mix homogeneously after 10 pounds of sterilizations in 10 minutes of high pressure and is got chlosma rhzomorph liquid.
A kind of chlosma emulsion agent for the treatment of chloasma is made up of described chlosma rhzomorph liquid and excipient, and described excipient is for adding 20ml propylene glycol, the formulated 100ml emulsion agent of 10ml glycerol.Wherein, the every ml of bacteria suspension contains about 88 hundred million of chlosma.
Claims (7)
1, a kind of chlosma preparation for the treatment of chloasma, make by chlosma rhzomorph and adjuvant:
Described preparation is any preparation for external application to skin, and the every ml of described chlosma rhzomorph bacteria suspension contains 30～90 hundred million of chlosmas.
2, the chlosma preparation of treatment chloasma according to claim 1, described adjuvant is skin usual excipients and additive.
3, the preparation method of the chlosma preparation of the described treatment chloasma of arbitrary claim in the claim 1～2 comprises chlosma rhzomorph and adjuvant are mixed.
4, the preparation method of the chlosma preparation of the described treatment chloasma of claim 3, the preparation of described chlosma rhzomorph may further comprise the steps: the collection cultivation of chlosma and the preparation of bacterium liquid;
Described collection cultivation comprises by the culture medium of following parts by weight of component makes: get Semen Glycines powder and add the mid-refrigerator overnight taking-up of distilled water; Filter, add peptone 1 in 100 parts in the filtrate: agar 1.8: the ratio of glucose 4 adds mentioned component; Culture medium is made in the heating and melting sterilization; Inoculation was cultivated 5～7 days for 37 ℃;
The preparation of described bacterium liquid comprises that the chlosma lawn of getting cultivation is dissolved in the normal saline, makes suspension; And/or, the lawn of cultivating is inoculated in the fluid medium, put in 37 ℃ of shaking table culture medium, cultivated 5～7 days, make bacteria suspension; Above-mentioned bacteria suspension filtration sterilization promptly.
5, the preparation method of the chlosma preparation of the described treatment chloasma of claim 4, the preparation of described chlosma rhzomorph may further comprise the steps: described collection cultivation comprises by the culture medium of following parts by weight of component makes, and gets 1000 parts of mid-refrigerators of distilled water of 40 parts of addings of Semen Glycines powder and crosses liquid taking-up next day; With filter paper or 8 layers of filtered through gauze, add mentioned component in 100 parts of ratios that add 1 part of peptone, 1.8 parts in agar, 4 parts of glucoses in the filtrate; Heating and melting, packing, 10 pounds of sterilizations in 10 minutes, it is standby to put the inclined-plane; To prepare the liquid branch and pack in the conical flask, it is standby to sterilize, and makes solid medium;
Describedly draw materials and cultivate to chlosma being inoculated in the surface of above-mentioned solid medium, cultivated 5～7 days for 37 ℃;
Behind the described bacteria suspension sterilization, filter standby; Describedly put sterilization behind 37 ℃ of shaking table culture medium culturings, cross the elimination sediment, standby.
6, a kind of application of chlosma preparation aspect preparation treatment chloasma medicine for the treatment of chloasma as claimed in claim 1.
7, a kind of application of chlosma preparation aspect preparation treatment chloasma cosmetics and skincare product for the treatment of chloasma as claimed in claim 1.
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| CNB031380433A CN1233330C (en) | 2003-05-30 | 2003-05-30 | Extraction of Malassezia for treating chlosma, said prepn. and its prepn. method and application thereof |
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| CNB031380433A CN1233330C (en) | 2003-05-30 | 2003-05-30 | Extraction of Malassezia for treating chlosma, said prepn. and its prepn. method and application thereof |
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| CN1233330C true CN1233330C (en) | 2005-12-28 |
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