• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The invention relates to a novel microorganism, namely the strain Trichosporon Kashiwayama, a culture product or culture liquid of said microorganism, sterile liquid, sterile filtrate, sterile supernatant of said culture liquid, or concentrate or dry product thereof, a process for the preparation thereof, a dermatic medicament having curative effect on skin disorder or a cosmetic consisting solely of or comprising as a main ingredient the above product, liquid, filtrate, supernatant, concentrate or dry product.
    公开号:SU1327791A3
    申请号:SU813278848
    申请日:1981-04-30
    公开日:1987-07-30
    发明作者:Касиваяма Синеи
    申请人:Синей Касива ма (JP)；
    IPC主号:
    专利说明:

    This invention relates to microbiology, in particular to the microbiological production of drugs.
    The purpose of the invention is to develop a method for obtaining a substance used for stimulating reparative co-processes, and identifying a producer strain capable of producing a substance with a stimulating effect on the reparative processes of the skin.
    The Trichosporon kashiwayama strain was deposited in the culture collection of the Research Institute of Fermentation, Japan, under the number FERM-P 4821, and is characterized by the following features.
    Morphological signs.
    The shape and size (liquid-environment on malt extract MM).
    After cultivation for 3 days, cells having a rectangular or oval are observed. the shape and size (3-4) X (2-20) µm, after culture for 5 days and more, the shape of the cells becomes irregular and, finally, the cells become mycelioid.
    Formation of pseudomycelium (culture on a glass slide, culture medium - agar with potato extract) with
    The cells are pseudohyphalic or pseudo mycelial; they have characteristic zigzag arthrospores, but without conidia. I
    Ascospore cells are not
    Ascospores have
    Ballistospores, Ballistospores cells do not have.
    Cultural signs.
    Colony on agar culture medium with streaking (agar culture medium on malt extract of MChO).
    Dim, white, hairy and a cusp of the surface, very soft, without rises, but with a ciliary peripheral part.
    Film on liquid culture medium (liquid culture medium on malt extract MCh).
    Bela, hairy, flaky and durable film.
    At 50% (May / May) glucose agar growth is absent, at 60% (May / May) agar medium with yeast extract does not grow.

    Physiological signs.
    Fermentation of carbohydrates.
    Glucose, galactose, sucrose, lactose, trehalose, raffinose and inulin do not ferment.
    Assimilation of carbon sources.
    B-glucose +, B-hapacose +, maltose-, raffinose-, sucrose-, lactose-, sodium lactate +, B-xylose +, erythritol-, 1) -mannit +, 1.-sorbose +, inositol-, trehalose -, D-arabinose-, L-arabinose-, sodium succinate +, sodium citrate +, sodium acetate +, inulin-, soluble starch-, ethanol +, glycerin +, cellobiose-, (+: , -: not used).
    Use of nitrates: not used.
    Use of nitrite: not used. enjoy
    Use of ztilamine hydrochloride: not used.
    Decomposition of glucoside. (arbutin) -: decomposes.
    Formation of starch analogues: not formed.
    Pigment formation: not formed.
    Formation of esters: are formed.
    Litmus milk reaction: does not coagulate.
    Vitamin Requirement: Not Required.
    Salt resistance: critical concentration 6-8% (May / vol).
    The optimum temperature is 25 C.
    The optimum pH value is 5.0.
    Urea degradation: does not degrade.
    Growth at 37 ° C: not growing.
    Acid formation from glucose: not formed.
    Cycloheximide resistance: resistant.
    Fats and oils decompose.
    Causes when cultured on a medium consisting of 0.3% (May / vol) glucose, 0.5% (May / vol.) Skim milk and 0.05% (May / vol) yeast extract , at pH 4.0–6.0 at 20–30 0 for 20 h, the biologically active substance, and the filtrate containing this substance, has the following properties.
    The cell free purified filtrate of the supernatant of the culture product contains small amounts

    The amount of amino acids, saccharides and lipids. When conducting a quantitative analysis of the filtrate, it turned out that the total nitrogen is 0.005 0.015%, and the total phosphorus - 0.004 -0.014%.
    Character and condition.
    The filtrate is a clear liquid with a color from colorless to slightly yellowish and has a faint odor.
    Samples for confirmation.
    20 ml of the filtrate are evaporated in a water bath to reduce the volume to approximately 50 ml. Then 1 ml of the hydrin sample solution is added to the concentrate. After the mixture is heated for 30 mn, the liquid turns violet.
    5 MP of nitric acid is added to the concentrate obtained by conducting the evaporation according to a known method, and the mixture is boiled for 20 minutes. After cooling the mixture, it is neutralized with a 10% sodium hydroxide solution. Then 2 ml of an ammonium-molybdate sample solution is added to the mixture. If the mixture is heated, the liquid becomes yellowish. If a 10% sodium hydroxide solution is added to the liquid, it becomes colorless.
    To 5 ml of the filtrate was added 10 mg of indole and 2 ml of hydrochloric acid, and the mixture was shaken for a sufficient time. When the mixture is then heated for 10 minutes, the liquid turns red.
    Physical characteristics:
    Specific gravity
    () 1,000-.1,015
    Coefficient pre1, 330-1,340
    4.5-5.5
    Residue after evaporation,% 0.5-2.0 Water content (according to the method of drying at atmospheric pressure)% 98.7 Protein (coefficient 6.25) Traces Lipid content (according to the extraction method in the Soxhlet apparatus),% 0.2 Content fiber% O
    "
    . fragmentation (p) pH (25 ° C).
    . five

    Q
    - jo
    25 so
    with
    40
    45
    50
    55
    Ash content,% 0.1 Content of sugar,% 1.0 Total dry matter content,% (in terms of water) 1.3 Content of nitrogen in amine-form, mg.% 10 Acidity by titration, MP number 1 n alkali , titrated to neutralize 100 g sample 0.6
    The nitrogen content in the form of volatile bases, m.% 1
    Mouse content (in the form of ASjOj) Not revealed (detection limit 0.1 h / min
    Heavy metal content (as Pb) Not found
    (detection limit of 1 h / min) Total mercury content Not detected
    (detection limit
    0.01 h / min)
    Cadmium content Not detected (detection limit
    0.01 h / min) Calcium content, mg.% 14.7
    Phosphorus content, mg.% 9.8 Formic acid content Not detected (detection limit 0.1%)
    Acetic acid content was not detected (detection limit 0.01%). Butyric acid content was not detected (detection limit 0.01%) Lactic acid content,% 0.06
    Lemon content
    acid%
    Number of living cells
    E.coli
    Staphylococcus aureus
    Number of Fungus Number of Yeast Propionic Acid Content
    vitamin B content
    vitamin B content, mg.%
    Ummarna content of itmin C
    vitamin B content
    Vitamin B content
    0.02
    Less than 30 cells per 1 ml / - /
    / - /
    / - // ml / - // ml
    fO
    Sorbic acid content
    Benzoic acid, g / kg
    P-hydroxybenzoic acid ester
    Not revealed (detection limit 5 0.02 g / kg)
    Not revealed (pre detection 20% 0.005 g / kg)
    25
    0.42
    Not revealed (detection limit .30 0.005 g / kg)
    Not revealed (detection limit 0.01 mg.%)
    0.02
    35
    40
    P
    Not revealed (detection limit 2 mg.%)
    not detected (detection limit not 5 µg.%) Not detected (detection limit-. nor 0.05 µg.%)
    45
    50
    55
    Pantogenic acid content
    mg.%
    Choline content
    O
    five
    0
    five
    0
    35
    0
    45
    0
    55
    Folic acid content
    Niacin Content
    Total Carotene Content
    PH value Amino acid composition of the amino acid in 100 g of the sample (cystine: tartaric acid oxidation method, tryptophan: analysis using microorganisms)
    Arginine
    Lysine
    Histidine
    Phenylalanine
    Tyrosine
    Leucine
    IsoleucineMethionine
    Valin
    Alanya
    Glycine
    Proline
    (detection limit 0.03%)
    Not detected (detection limit 1 µg.%) Not detected (detection limit, 0.03 mg.%)
    Not revealed (detection limit 0.02 mg.%) 4.9
    less than 0.01 less than 0.01 less than 0.01 Less than 0.01 Less than 0.01 Less than 0.01 Less than 0.01 Less than 0.01 Less than 0.01 Less than 0.01 Less than 0.01 Less than 0.01 Glutamic acid Less than 0.01 Serin Less than 0.01
    Threonine Less than 0.01
    Aspartic acid Less than 0.01 Tryptophan Less than 0.01 Cystine Less than p, 01 A cell-free, sterile supernatant can be used as a dermatological drug apparatus and without concentration. However, it is preferred to concentrate the cell-free sterile supernatant. In addition, it is possible to use a concentrate with a high degree of concentration exceeding 25 or a dry product. A similar concentrate having a high degree of concentration or
    dry product, can be used - As a dermatological drug. In addition, a dermatological preparation can be obtained by mixing a similar concentrate, strong concentrate or dry product with a cream, ointment base or other carrier.
    The results of the toxicity test using chicken embryo show that the cell-free sterile concentrate (concentration level 4) is not toxic. The results of an allergic skin test on the human body and a test for continuous irritation of human skin show that the concentrate is completely non-toxic to human skin. The results of the rabbit mucous membrane irritation test show that the concentrate is not an irritant at all. The results of the antibiotic test show that the concentrate does not contain antibiotic substances.
    The dermatological drug has a healing effect on skin spots, cracked skin, contact dermatitis and burned skin.
    Sample for the cure of skin spots.
    621 women with skin spots on the face, with the exception of patients with liver disease or gynecological disorders, and pregnant women admitted to a dermatological hospital for treating skin spots are selected. After washing the face, the skin locks otdermatitis, presumably caused by reasons not related to this treatment. Percentage of patients for whom significant or intelligence is observed
     5% improvement, 80%. In addition, in 60% of 365 women, in whom the condition has significantly improved, the spots on the skin have completely disappeared.
    0 The test for the healing of cracks in the skin and small wrinkles.
    For the bath sample, 962 women are selected from among the enrollments in dermatol.
    J5 hospitals (150 women aged 25–30 years old, 218 women aged 30–35 years old - 248 years old 35–40 years old, 172 women age 40–40 years old, and 174 women age
    20 those over 45 years). After washing the skin of the face, the skin pores are opened with ozone and steam, gauze impregnated with a concentrate is placed tightly on the skin of the face (degree of concentration 2)
     The cell-free supernatant and gauze are covered with a Saran-Wrap film. Irradiation with infrared rays is carried out for 15-20 minutes to improve the penetration of the concentrate into the skin. After one treatment, the cracked skin was found to significantly improve in 698 women (73%), comparatively improved in women (16%), the condition did not change in 72 women (7.2), and the condition did not worsen in any of women, 36 women remain (4%). Considering the fact that skin cracking develops in the final
    they are affected by ozone and steam, and on a 40 account in fine wrinkles, it can be considered that the concentrate, which is
    gauze impregnated with a concentrate (degree of concentration 2) of a cell-free supernatant. The applied gauze is covered with Saran-Wrap film and infrared rays are applied to improve the penetration of the concentrate into the skin. This treatment was carried out once a week and continued for 3 months. It was found that the condition improved greatly in 363 women (58.4%), the condition improved comparatively in 122 women (19.8%), no improvement in 133 women (21.4%), the condition worsened in one woman (0.01%) and contact dermatitis occurred in the other two women (0.03%). Contact
    dermatitis, presumably caused by reasons not related to this treatment. The proportion of patients for whom significant or moderate improvement is observed is 80%. In addition, 60% of 365 women in whom the condition has improved significantly, the spots on the skin have completely disappeared.
    The test for the healing of cracks in the skin and small wrinkles.
    For the bath sample, 962 women are taken from the number of admissions to dermatological hospitals (150 women aged 25–30 years, 218 women aged 30–35 years, - 248 aged 35–40 years, 172 women aged 40–50 years and 174 women over the age of 45). After washing the skin of the face, the pores of the skin are opened with ozone and steam, gauze impregnated with concentrate is tightly applied to the face (degree of concentration 2)
    The cell-free supernatant and gauze are covered with a Saran-Wrap film. Irradiation with infrared rays is carried out for 15-20 minutes to improve the penetration of the concentrate into the skin. After one treatment, the cracked skin was found to significantly improve in 698 women (73%), comparatively improved in women (16%), the condition did not change in 72 women (7.2) and the condition did not worsen in any of the women. 36 women remain (4%). Considering the fact that skin cracking develops in the final
    effective in terms of treating skin cracking, has a preventive effect, preventing the formation of fine wrinkles.
    Test for therapeutic effect on contact dermatitis.
    A 21-year-old woman was selected, suffering from a rash and burning on both eyelids on the eyelids on the following 1st day after the change of eye shadow company A and on eye shadow of company B, the diagnosis is contact dermatitis. A dermatologist applied to the affected areas a gauze soaked in a concentrate (concentration 2) of the cell-free supernatant. No steroid used.
    five
    913277911.0
    drugs, anti-inflammatory mix fermentation and antihistamine substances.Patsi-aerotank. Then the cultivation of the equipment is presented to about 20 ml of condensate, with a concentrate for 70 hours, the concentrate is applied for 30 minutes of aeration and mixing. Twice a day, i.e. in the morning and in the evening, 5 completion of the fermentation culture-. After 48 hours after the start of treatment, the oral fluid is passed through the burning sensation, and after 96 hours the filter apparatus of the model after the start of treatment, take substantially 293-16 manufactured by the company (Naiveu disappeared. After 1 week rash not gay Sekukhin Koge KK (filtering is completely; O on the GL-90 paper produced by Firprob for the treatment of burns, We Toe Rosie, 0.5 µm) and through the filterWoman, 47 years old, having a burn 293-4 resulting from the scalding of the production of the same firms s (membranes with hot water, on the outer part of the filter-TM-2 manufactured by
    the left forearm (II degree burn; Toi Rosi, 0.45 µm) for removal) with the eruption and swelling of particulate matter and cells. Obtained from the palm, as well as small, the inferior filtrate is concentrated in the corresponding manner, turned to a dermatologist with the method of using membranes after 1 h after scalding. The reverse osmosis porous in the apparatus of the mo- tional site is immediately cooled20 with the production of the company Bayoindzhi- running water for the affected part of Co. so that the volume of reduction is applied to gauze impregnated up to 200 liters. The concentrate obtained (degree of concentration 2) is sterilized using a heat-cell sterile supernatant of a dish-type liquid exchanger and is applied with a first- (manufactured by Ivan Kikay Koge gantnoy paper. Then on the affected KK), sterile Packed in a stretch of dressing. glass containers and stored in a good- After 24 hours, it was found that small loader (5 C). the bubbles have completely disappeared, although PRImeR 2. Sterilized
    The rash and swelling remained and-30 concentrate obtained in Example 1, the patient stopped complaining of freeze-drying and the resulting powder was mixed with sterile ointment. A 30-year-old man who had a burn in a base with an ointment to burn hot water of the skin.
    on the back of the right leg (burn C35) PRI me R 3. Cosmetic
    degrees) with loosening and swollen - the preparation is obtained by injecting 1 g with chicken-sized meat, treated with concentrated concentrate, obtained with concentrate (degree of concentration in example 1, in cosmetic 2) with cell-free sterile-base, consisting of 1 g of stearic supernatant according to Method 40, 7 g of beeswax, ke, as described above. After 24 hours with 4 g of self-emulsified glycerol, the swelling disappeared, although the precipitate of osteostearate, 30 g of liquid paraffin melted, and the patient stopped feeding — 0.1 g of water lanolin and 40 g of purified parasite, and plasticizing the mixture
    t,;; in accordance with traditional methods, concentrate turns out to be wild. The composition is poured and falls heavily and is highly therapeutic. " v t- effect in relation to burns. forged into ampoules, and the packed amp examples are mixed for sale,
    fermentation aeration tank with a capacity of 4. Example 4. A medicinal dose of 1000 liters is loaded with 800 liters of cultured skin skin treatment medium described above, obtained by administering suitable amounts of composition, and after sterilization by heating and pyridoxin hydrochloride and natural cooling of Y-ω-ethanol, allatoin, DL-i-tokoetu culture medium is inoculated with (vitamin E) and mint oil
    ; „EZ TO THE COMPOSITION obtained in example 3. by a seed formed by
    权利要求:
    Claims (2)
    [1]
    by pre-cultivation Formula of the invention at 28 ° C for 30 h in 10 l1. Strain Trichosporon kashiwayama
    of the same culture medium in small FEKK-P 4821 (Collections of cultures
    11 132779112
    Ferm, Japan) producing a substance that produces glucose Of37, (May / vol)
    having a stimulating effect of 0.5% milk (ma.ob.), yeast
    on the reparative processes of the skin. extract 0.05% (ma.ob.), at pH
    [2]
    2. The method of obtaining a substance, about 4.0-6.0 and a temperature of 20-30 ° C, under aerodynamic stimulating action conditions, then separates the biopreparation processes of the skin, closure in sterile conditions and the resulting trichosted sterile filtrate strain with the necessary kashiwayama ferm-p 4821 growth is concentrated and / or dried in a nutrient medium, containing.
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    同族专利:
    公开号 | 公开日
    JPS5636419A|1981-04-09|
    HU186767B|1985-09-30|
    FI802690A|1981-03-02|
    ES494636A0|1981-08-16|
    ZA805361B|1981-09-30|
    DK157035C|1990-03-26|
    DK157035B|1989-10-30|
    EP0024738A2|1981-03-11|
    EP0024738A3|1981-10-07|
    IN153038B|1984-05-26|
    EP0024738B1|1984-08-22|
    PH22011A|1988-05-02|
    DE3069020D1|1984-09-27|
    FI68079B|1985-03-29|
    US4503041A|1985-03-05|
    US4554161A|1985-11-19|
    MX6444E|1985-05-31|
    CA1176586A|1984-10-23|
    UA6318A1|1994-12-29|
    JPS6143033B2|1986-09-25|
    FI68079C|1985-07-10|
    CS247055B2|1986-11-13|
    DE24738T1|1983-04-14|
    DK194981A|1981-05-01|
    AR228434A1|1983-03-15|
    WO1981000723A1|1981-03-19|
    AT9100T|1984-09-15|
    ES8107299A1|1981-08-16|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    MD210C2|1979-09-01|1995-11-30|Toyohide Kashiwayama|Strain Trichosporone Kashiwayama which is producing the substance possessing stimulus action on the skin reparative processes and method of preparation thereof|FR1443067A|1963-06-28|1966-06-24|Process for the preparation of powders containing products with high activity, resulting from the exchange of substances, from microorganisms|
    JPS50145577A|1974-05-16|1975-11-21|US4992534A|1987-08-12|1991-02-12|Otsuka Pharmaceutical Co., Ltd.|3'-O-,5'-O-derivatives of 2'-deoxy-5-fluorouridine|
    FR2679140B1|1991-07-19|1993-10-15|Oreal|DEPIGMENTING COSMETIC OR DERMATOLOGICAL COMPOSITION CONTAINING ARBUTOSIDE DERIVATIVES.|
    US5578296A|1993-02-08|1996-11-26|Kabushiki Kaisha Kobe Seiko Sho|Decomposition of melanin using a culture of basidiomycetes fungus|
    AU6012594A|1994-01-28|1995-08-15|Sergei Borisovich Alexeev|Strain of trichoderma harzianum rifai, process for obtaining l-lysine-alpha-oxidase as an inhibitor of viral and bacterial activity, immunomodulator and skin healing agent|
    US5866142A|1995-07-20|1999-02-02|Riordan; Neil H.|Skin treatment system|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    JP54112294A|JPS6143033B2|1979-09-01|1979-09-01|LV930944A| LV5534A3|1979-09-01|1993-06-30|Strain - Producer and Substance Has Stimulating Effect on Stimulating Effects on Skin Reparative Processes|
    LTRP1025A| LT2525B|1979-09-01|1993-09-21|FRICHOSPORON KASHIWAYAMA MICROORGANISM CAMERA-MATERIALS CONTAINING STIMULATIVE EFFECTS FOR DIAGNOSTIC EXPOSURE PROCESSES, MANUFACTURING AND RECEIVING MATERIALS|
    MD94-0230A| MD210C2|1979-09-01|1994-06-01|Strain Trichosporone Kashiwayama which is producing the substance possessing stimulus action on the skin reparative processes and method of preparation thereof|
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