WO2005059114A1 - Composition de graisse favorisant la croissance de cellules animales, milieu de culture de cellules animales contenant ladite composition et preparation cutanee externe - Google Patents

Composition de graisse favorisant la croissance de cellules animales, milieu de culture de cellules animales contenant ladite composition et preparation cutanee externe Download PDF

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WO2005059114A1
WO2005059114A1 PCT/JP2004/018575 JP2004018575W WO2005059114A1 WO 2005059114 A1 WO2005059114 A1 WO 2005059114A1 JP 2004018575 W JP2004018575 W JP 2004018575W WO 2005059114 A1 WO2005059114 A1 WO 2005059114A1
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Prior art keywords
composition
oil
mass
fat
cell growth
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PCT/JP2004/018575
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English (en)
Japanese (ja)
Inventor
Toshitsugu Shimauchi
Toshiaki Nakajima
Akihiro Kondo
Shuichi Hiyamuta
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Idemitsu Kosan Co., Ltd.
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Priority claimed from JP2003419616A external-priority patent/JP2005179211A/ja
Application filed by Idemitsu Kosan Co., Ltd. filed Critical Idemitsu Kosan Co., Ltd.
Publication of WO2005059114A1 publication Critical patent/WO2005059114A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67—Vitamins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/37—Esters of carboxylic acids
    • A61K8/375—Esters of carboxylic acids the alcohol moiety containing more than one hydroxy group
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
    • C12N5/0629—Keratinocytes; Whole skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/36—Lipids

Definitions

  • Animal cell growth promoter oil composition animal cell culture medium containing the composition, and skin external preparation composition
  • the present invention relates to an animal cell growth promoting oil / fat composition comprising ⁇ monolinolenic acid (hereinafter, abbreviated as GLA) ⁇ 7-linolenic acid glyceride having animal cell growth promoting ability, and
  • GLA ⁇ monolinolenic acid
  • the present invention relates to a medium for culturing animal cells comprising the oil / fat composition.
  • the present invention relates to a skin external preparation composition
  • a skin external preparation composition comprising, as an active ingredient, a fat or oil composition containing ⁇ -linolenic acid or ⁇ -linolenic acid dalyseride having an ability to promote animal cell growth.
  • Animal cell culture technology is widely used, for example, for cell fusion, production of monoclonal antibodies, production of vaccines, and the like. Is a biological technology.
  • Animal cells do not grow in a medium containing only nutrients such as amino acids, sugars, salts, etc. In order to grow animal cells, a cell growth agent other than mere nutrients is required.
  • 5-aminolevulinic acid for example, see Patent Document 1
  • RU substance for example, see Patent Document 2
  • 5-hydroxy-2,7-dimethoxy 6-methyl-1,4-naphthoquinone See, for example, Patent Document 3
  • Panax ginseng extract see, for example, Patent Document 4
  • Animal cell promoters such as toglycogen (for fibroblasts) (for example, see Patent Document 5) and sugar esters of sugars (monosaccharides and disaccharides) and fatty acids (for example, see Patent Document 6) have been developed.
  • these animal cell promoters have problems such as difficulty in production, high production cost, and unstable quality, and the development of an inexpensive animal cell promoter with excellent quality stability is expected. It is rare.
  • retinol polyhydroxyacid is effective in repairing wrinkles as a substance that has the effect of improving skin aging symptoms such as wrinkles and a substance that prevents skin aging and inflammatory effects due to sunlight and ultraviolet rays.
  • ⁇ -hydroxy acids dalicholic acid and lactic acid, which have particularly effective effects, are lipophilic, and if the compounding amount is poor in percutaneous absorption, undesired side effects such as skin irritation may occur.
  • retinol is easily oxidized when exposed to air, and is usually used as an ester with fatty acids S. Since this fatty acid ester is fat-soluble, it is difficult to mix it with a water-soluble base material. there were.
  • wounds surgical incisions, wounds or ulcers in the gastrointestinal tract, abrasions, lacerations, amputations, so-called bed sores, sores, infections and other damage to superficial tissues
  • an agent that stimulates or promotes the process of differentiation and proliferation of cells involved in the healing process of a wound which depends on the formation of cells, is effective.
  • Extracts such as aloe, antibiotics, anti-inflammatory drugs, kallikrein, adenine, nicotinic acid, allantoin, vitamin A, zinc, and cAMP derivatives as active ingredients that exhibit wound healing effects (see, for example, Patent Document 7)
  • b-EGF cell growth factor
  • the above-mentioned active ingredient may not be able to obtain a sufficient effect, and there are problems that production is difficult, the production cost is high, and the active ingredient is chemically unstable. Excellent, has the effect of improving or healing skin symptoms There is a need for a skin external preparation that does.
  • Patent Document 1 JP-A-5-49472
  • Patent Document 2 JP-A-5-97685
  • Patent Document 3 JP-A-5-137569
  • Patent Document 4 JP-A-6-239759
  • Patent Document 5 JP-A-11-255657
  • Patent Document 6 JP-A-2003-52366
  • Patent Document 7 JP-A-63-107935
  • Patent Document 8 JP-A-3-106823
  • the present invention has been made in view of the above circumstances, and provides an oil / fat composition which is inexpensive, has excellent quality stability, and has an ability to promote the growth of animal cells, and a medium for culturing animal cells containing the oil / fat composition. It is intended for that purpose.
  • Another object of the present invention is to provide a skin external preparation composition which is inexpensive, has excellent quality stability, and has an effect of improving or healing skin symptoms.
  • the present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, have found that the above object can be achieved by a fat or oil composition containing a specific substance having an animal cell growth promoting ability. I found it.
  • the present invention has been completed based on powerful knowledge.
  • the present invention relates to an oil / fat composition containing triglyceride, diglyceride, monoglyceride and free fatty acid, wherein the oil / fat composition contains one or more selected from ⁇ -linolenic acid and ⁇ ⁇ -linolenic acid glyceride. Is provided.
  • the present invention also provides an animal cell culture medium containing the animal cell growth promoter oil / fat composition.
  • the present invention is a triglyceride, diglyceride, a fat composition comprising a monoglyceride and free fatty acids, I - linolenic acid and I - to provide a skin external composition containing one or more upper selected from linolenic acid glyceride Is the thing The invention's effect
  • an animal cell growth promoter oil / fat composition which is inexpensive and has excellent quality stability, and an animal cell culture medium containing the animal cell growth promoter oil / fat composition can be obtained.
  • a skin external preparation composition which is inexpensive, has excellent quality stability, and has an effect of improving or healing skin symptoms can be obtained.
  • the fatty acids constituting triglycerides, diglycerides, and monoglycerides which are components of the animal cell growth promoter oil / fat composition or skin external preparation composition of the present invention are not limited, for example, saturated fatty acids having 2 to 24 carbon atoms and unsaturated fatty acids.
  • Preferred fatty acids include caprylic acid, capric acid, myristic acid, myristoleic acid, pentadecenoic acid, palmitic acid, panolemitrenic acid, stearic acid, arachidic acid, behenic acid, ligrenolic acid, serotinic acid, oleic acid, and linoleic acid.
  • Monooleic acid monolinolenic acid, ⁇ -linolenic acid, octadecatetraenoic acid, eicosenoic acid, eicosadic acid, eicosatrienoic acid, eicosatetraenoic acid, arachidonic acid, eicosapentaenoic acid, docosenoic acid, Docosagenic acid, docosapentaenoic acid, docosahexaenoic acid and the like.
  • Glyceride is not particularly limited, and is preferably a plant or microbial-derived glyceride, because it can be used in a chemically synthesized product, a plant or animal-derived, or a microbial-derived.
  • the content ratio of triglyceride, diglyceride, and monoglyceride contained in the animal cell growth promoter oil composition or the skin external preparation composition of the present invention is based on the total amount of glyceride from the viewpoint of preventing variation in the animal cell growth effect. , Respectively, preferably 10 to 79% by mass, 20 to 70% by mass, and 1 to 70% by mass.
  • the content of triglyceride, diglyceride and monoglyceride is more preferably 10 to 70% by mass, 25 to 60% by mass and 3 to 65% by mass, respectively, based on the total amount of glyceride. Most preferably, they are 10-65% by mass, 30-60% by mass and 5-60% by mass, respectively, based on the total amount of glyceride.
  • Glycerides having such a composition can be produced by a known method. For example, it is produced by a method of inducing triglyceride type fats and oils into diglycerides and monoglycerides using a chemical catalyst or an enzyme catalyst (for example, see JP-A-2000-236888 and JP-A-2000-333688). The ability to do S.
  • GLA which is an active ingredient of the oil / fat composition for animal cell growth promoter or the external preparation for skin of the present invention, may exist not only as a free fatty acid but also as an ester.
  • GLA content refers to a free fatty acid obtained by hydrolyzing glyceride from all GLA derivatives in the oil / fat composition of the present invention, a fat / oil composition for animal cell growth promoter, or an external preparation for skin. It is calculated from the total free fatty acids present in the composition. For example, when GLA is present as a ester, the amount of free GLA obtained by hydrolyzing this ester with kafunka may be measured.
  • the content of GLA in the animal cell growth promoter oil / fat composition may be appropriately adjusted according to the type of animal cell to be subjected to growth promotion.
  • the amount is preferably 5% by mass or more based on the total amount of fatty acids and free fatty acids constituting glyceride. It is more preferably at least 10% by mass, and even more preferably at least 20% by mass.
  • the content of GLA it is preferably 98% by mass or less from the economical point of view, even if the content exceeds 98% by mass, even if the effect of increasing animal cell proliferation is not further improved.
  • the content of GLA in the skin external preparation composition may be appropriately adjusted according to the dosage form and type of the skin external preparation composition.
  • the content is preferably 5% by mass or more based on the total amount of fatty acids and free fatty acids constituting glyceride, from the viewpoint of preventing the healing effect from being varied depending on the dosage form and the intended use. It is more preferably at least 10% by mass, and even more preferably at least 20% by mass.
  • the content of GLA it is preferably 98% by mass or less from the economical viewpoint, even if the content exceeds 98% by mass, the healing effect is not further improved.
  • GLA can also be obtained by chemically synthesizing, plant seeds such as borage, evening primrose, and saxifraga, algae such as Spirulina, Cunninghamella, Mortierella, and Mucor. Microbial power of a genus or the like can also be obtained.
  • the obtained GLA is a derivative, it may be subjected to hydrolysis treatment or transesterification treatment which may be used as it is.
  • the animal cell growth promoting oil / fat composition or skin external preparation composition of the present invention is produced by mixing glyceride and GLA oil / fat at the above-mentioned predetermined ratio by a known mixing method. You can. Further, the animal cell growth promoting oil or fat composition or the skin external preparation composition of the present invention extracts a GLA-containing microbial fat inside a microorganism or a microbial diluting fat from a GLA-containing microorganism, and converts the GLA-containing fat or oil into It can be obtained by hydrolysis or transesterification.
  • the GLA-containing fats and oils can be obtained by culturing a microorganism capable of producing GLA-containing fats and oils by a conventional method.
  • examples of the microorganism having the ability to produce GLA-containing fats and oils include various microorganisms such as filamentous fungi, yeast, and algae.
  • microorganisms having the ability to produce gamma-linolenic acid-containing fats and oils include microorganisms belonging to the genus Mortierella described in JP-A-60-168391 and JP-A-63-283589. And the like, and microorganisms belonging to the genus Rhizopus described in JP-A-63-133994 and the like. More specifically, as a microorganism belonging to the genus Mortierella, for example,
  • Mucor circinelloides HUT1121 (FERM P-9359) @ Mucor * Javanitas (Mucor javanicus) HUTl 162 (FERM P-9360).
  • the medium for culturing the microorganism may be a carbon source, a nitrogen source, inorganic salts, and, if necessary, an amino acid suitable for the growth of the microorganism, as long as the microorganism can grow well and produce a desired lipid. What contains the component of is used.
  • the carbon source saccharides such as darcos, starch, molasses, and organic acids and sodium acetate can be used, and saccharides such as gnoleose are particularly preferable.
  • the nitrogen source include ammonium salt, yeast extract, corn steep liquor, peptone, and the like.
  • the inorganic salts include magnesium salts, calcium salts, phosphates, iron salts, and copper salts.
  • culturing In culturing, if the total amount of carbon source, nitrogen source or phosphate is added to the medium from the beginning, it may adversely affect the growth of microorganisms. , Which can improve the productivity of ⁇ -linolenic acid.
  • Other culturing conditions such as temperature and culturing time, are preferably set in consideration of the medium composition, the target content of ⁇ -linolenic acid, and the productivity of lipids.
  • the temperature is usually about 20 to 35 ° C, preferably 25 to 30 ° C, and ⁇ is usually about 3 to 7, preferably 3.5 to 6.5, usually about 72 to 240 hours, preferably about 96 to 168. It's time to go.
  • GLA-containing lipids are usually accumulated in microbial cells
  • the culture solution is subjected to solid-liquid separation by a conventional method to obtain cells containing GLA-containing lipids.
  • these cells can be used as they are, but in order to further extract and purify fats and oils, the bacteria are obtained by the methods described in the Bligh & Dyer method, the Folich method, and JP-A-57-144986. By crushing the body and extracting the solvent, the desired GLA-containing fats and oils can be obtained.
  • the GLA-containing fat or oil obtained as described above is subjected to hydrolysis treatment or transesterification treatment by a known method to obtain the animal cell growth promoter fat or oil composition or the external skin composition of the present invention. it can.
  • the animal cell growth promoter oil / fat composition or skin external preparation composition of the present invention is obtained by such a method, the content ratio of GLA in the animal cell growth promoter oil / fat composition or skin external preparation composition is determined as follows. A part of the accelerator or fat composition or the external composition for skin is determined, and after methylesterification, determined by gas chromatography (see JP-A-57-144986).
  • the animal cell growth promoter oil / fat composition of the present invention may contain other components as long as they do not inhibit the effects of the present invention.
  • Other components include, for example, the aforementioned fatty acids other than GLA.
  • GLA plant-derived or microorganism-derived glyceride
  • other components derived from a plant or a microorganism may be contained.
  • the animal cell growth promoter oil composition of the present invention comprises an adhesion-dependent cell such as epidermal keratinocytes, skin fibroblasts, melanocytes, small intestinal epithelial cells, human cervical cancer cells (Hela), and lung fibroblasts.
  • an adhesion-dependent cell such as epidermal keratinocytes, skin fibroblasts, melanocytes, small intestinal epithelial cells, human cervical cancer cells (Hela), and lung fibroblasts.
  • the animal cell growth promoting oil / fat composition of the present invention can be used without limitation as long as the use requires the growth of animal cells.
  • additives for animal cell growth media for example, additives for animal cell growth media, artificial skin agents (cell growth additives), pharmaceuticals, foods, cosmetic ingredients, etc. You can. Hereinafter, these uses will be described.
  • the animal cell culture medium that can be applied is not particularly limited as long as it is suitable for the animal cell to be grown, and may be a commercially available basic medium or a suitably modified one thereof.
  • Specific examples of the basic medium include MEM (Minimun Essential Medium) medium, Iskov medium, RPM11640 medium, Ham's F-12 medium, Dulbecco's modified Eagle medium, Dulbecco's modified MEM medium, and Iscov's modified Dulbecco medium with 10% FBS. And preferably Dulbecco's modified Eagle's medium. These media may be used alone or in combination of two or more.
  • the amount of the oil / fat composition of the present invention added to the culture medium may be appropriately adjusted depending on the target animal cell type.
  • the animal cell of the present invention is adjusted so that the concentration in the culture solution is about 0.001 to 100 mg / milliliter, preferably 0.01 to 10 mg / milliliter, and more preferably 0.011 mg / milliliter.
  • a growth promoter oil or fat composition is added.
  • the solvent used for preparing this solution preferably dissolves the animal cell growth promoter oil / fat composition of the present invention and is compatible with the culture solution.
  • examples of such a solvent include methanol, dimethyl sulfoxide, acetone, and the like, and preferably acetone. These solvents may be used alone or in combination of two or more. From the viewpoint of suppressing the effect of the added solvent on animal cells, the solvent is preferably used so as to be 2% by mass or less based on the total amount of the medium.
  • the animal cell growth promoter oil composition of the present invention it is possible to promote the early formation of artificial skin and increase of artificial skin by adding the animal cell growth promoter oil composition of the present invention to a culture medium for artificial skin production or applying or spraying the composition on cultured artificial skin. It can.
  • a solution or dispersion obtained by dissolving the animal cell growth promoter oil / fat composition of the present invention in an appropriate solvent may be applied or jetted.
  • the solvent those similar to the above can be used.
  • the solvent is preferably used in an amount of 2% by mass or less based on the total amount of the culture medium as described above.
  • the solvent can be applied or sprayed within a range that does not adversely affect the cultured human skin.
  • animal cell growth promoter oil / fat composition of the present invention When used as a component of pharmaceuticals, foods, and cosmetics, various effects due to promotion of animal cell growth can be expected.
  • a known method can be used for mixing the animal cell growth promoter oil / fat composition of the present invention with a drug or the like.
  • the skin external preparation composition of the present invention may contain other components generally used as skin external preparation components, as long as the effects of the present invention are not impaired.
  • Other components include, for example, purified water, alcohols, oily substances, anti-dandruff agents, bactericides, medicinal ingredients, preservatives, thickeners, astringents, humectants, powders, fragrances, and emulsion stabilizers. And a pH adjuster.
  • a plant-derived or microorganism-derived glyceride GLA is used, other components derived from a plant or a microorganism may be contained.
  • alcohols include ethanol, higher alcohols having a linear or branched alkyl or alkenyl group
  • oily substances include hydrocarbons such as liquid paraffin, vaseline, and solid paraffin; liquid lanolin, lanolin Lanolin derivatives such as fatty acids; silicone derivatives such as dimethylpolysiloxane, polyether-modified polysiloxane, and amino-modified polysiloxane, higher alcohol higher fatty acid esters, higher fatty acids, and fats and oils such as long-chain amidoamine having an alkyl or alkenyl group.
  • Animal and vegetable fats and oils such as mink oil and olive oil; Pharmaceutical ingredients include vitamins and the like.
  • Preservatives include parabens, and thickeners include water-soluble polymer compounds.
  • examples of the humectant include propylene glycol, glycerin, carbitol, 3-methyl-1,3-butanediol, and saccharides.
  • the dosage form of the skin external preparation composition of the present invention is not particularly limited and may be used as it is. Creams, emulsions, lotions, ointments, powders, haptics, powders, and drops , A patch or an aerosol can be applied in the form of a usual external preparation for skin.
  • the amount of the skin external preparation composition of the present invention may be appropriately adjusted depending on the use. For example, when formulated in cosmetics, the concentration in cosmetics can be usually about 0.001 lOOmg / milliliter, preferably 0.01 lOmg / milliliter, more preferably 0.011 lmgZmilliliter. To mix.
  • the concentration in the therapeutic agent is usually more than 0.001 mgZ milliliter (used as is).
  • the amount is preferably 0.01 to 500 mg / milliliter, more preferably 0.01 to 100 mg / milliliter.
  • the fatty acid composition of the raw material fats and oils and fat compositions used in the following Examples and Comparative Examples was methyl esterified by the method described in Japanese Patent Publication No. 577-144896 and measured by gas chromatography.
  • the constituent glyceride compositions of the oil and fat compositions of Examples and Comparative Examples were determined based on the Itroscan method (J. Jpn. Oil Chem. Soc., Vol. 35, No. 8, 625-631 (1986)).
  • Iatroscan TH_10 type manufactured by Jatron Co., Ltd.
  • TLC thin layer chromatograph
  • FID flame ionization detector
  • FID Flame ionization detector
  • the glyceride composition analysis was carried out using a part of the oil / fat composition by the above-described method, and the fatty acid composition was analyzed by gas chromatography after a part of the oil / fat composition was methylesterified.
  • the results are shown in Table 3.
  • C16: 0 means a fatty acid having 16 carbon atoms and no double bond
  • C18: 3 means a fatty acid having 18 carbon atoms and having 3 double bonds.
  • a 10% by mass aqueous solution of sodium hydroxide was added to the obtained decomposition reaction product to adjust the pH of the decomposition reaction product to 89, and then oil was extracted from the decomposition reaction product using 200 ml of getyl ether and washed with water. After dehydration, the ether was removed to obtain 87.9 g of an oil component (oil / fat composition).
  • Example 1 except that GLA-containing fats and oils having a fatty acid composition shown in Table 2 (manufactured by Idemitsu Kosan Co., Ltd., trade name: Daranoyl CS, GLA: 7.9% by mass) were used in Example 1, An oil / fat composition was obtained by the same enzyme treatment and extraction as in Example 1.
  • Table 3 shows the analysis results of the obtained fat and oil composition.
  • Example 1 an oil / fat composition was obtained by performing the same treatment as in Example 1 except that safflower oil containing no GLA acid (manufactured by SIGMA) was used instead of using the GLA-containing oil / fat.
  • Table 3 shows the analysis results of the obtained fat and oil composition.
  • Linoleic acid (c 18: 2 ) 11.7 13.1 17.1 74.9
  • Triglycerides TG 59.2 13.8 50.5 62.3
  • the proliferation test was performed by seeding normal human epidermal keratinocytes (manufactured by Sanko Junyaku Co., Ltd., Cryo NHEK-Neo: neonatal) at a cell density of 3.5 ⁇ 10 3 cells / well using a 96-well microphone plate.
  • Brett kit KMG basic medium manufactured by Sanko Junyaku Co., Ltd.
  • hEGF epidermal growth factor
  • insulin 0.5 ml Incubate for 6 days at 37 ° C, 5% CO
  • the number of cells was measured using a Premix WST-1 Cell Proliferation Assay System (manufactured by Takara Shuzo Co., Ltd.), and the absorbance at 450 nm was measured using a Pell Reader (manufactured by Seikagaku Corporation, model SK601).
  • the growth rate (%) when the oil / fat composition was added was calculated based on the number of cells in the test medium alone without the oil / fat composition of the present invention as a reference (100%). The results are shown in Table 4. You.
  • CH ⁇ cells cultured cells derived from Chinese hamster egg cells
  • CHO cells ATCC stock: CRL-11397 strain
  • CHO-S-SFM II medium GIBCO BRL
  • the proliferation test was performed as follows. That is, the oil and fat compositions obtained in Examples 13 and 13 and Comparative Example 1 were each added to a CHO-S-SFM II medium (manufactured by GIBCO BRL) at a concentration of 0.1 mgZZ to 10 ml of a test medium. 1 ⁇ 10 5 cells of CH ⁇ cells (ATCC storage strain: CRL-11397 strain) were suspended and added to the culture cell space in an integra cell line (CL-350, manufactured by INTEGRA BIOSCIE NCES). In addition, as a basal medium, 200 milliliters of a CHO-S-SFM II medium (GIBCO BRL) without the above-mentioned fat composition was added to the culture cell space.
  • a CHO-S-SFM II medium manufactured by INTEGRA BIOSCIE NCES
  • the above Integra cell line containing medium and CHO cells was added at 37 ° C in the presence of 5% CO.
  • the entire cell suspension was recovered. After the number of cells in this cell suspension was measured using a hemocytometer, the amount of IgG antibody produced in the culture supernatant was measured by quarantine fluorescence (ELISA). In addition, the number of cells and the amount of IgG antibody produced when the same test was performed using only the basal medium (CHO-S-SFM II medium (manufactured by GIBCO BRL)) not containing the oil and fat composition of the present invention were taken as reference (100%). The growth rate (%) and the IgG antibody production improvement rate (%) when the oil and fat composition was added were calculated. Table 5 shows the results.
  • Example 1 Glycerin (10 g), liquid paraffin (10 g) and white petrolatum (79 g) were dissolved while heating to 80 ° C, then cooled to 45 ° C, and the oil and fat composition lg obtained in Example 1 was added and mixed. Was quenched to prepare a uniform ointment. This was used as Example 1 ointment. Similarly, an ointment of Example 2, an ointment of Example 3, and an ointment of Comparative Example 1 were prepared using the oil and fat compositions obtained in Examples 2, 3 and Comparative Example 1.
  • the healing effect of these ointments was evaluated by the following method.
  • 20 women aged 50 to 60 who have aging symptoms (wrinkles) on the skin are divided into 4 groups of 5 women each, and the first group has Example 1 ointment and the second group has Example 2 ointment.
  • the ointment was used for one month, and the condition of wrinkles on the skin before and after use was evaluated according to the following evaluation criteria.
  • Table 6 shows the evaluation results. The numbers in Table 6 indicate the number of people who meet each evaluation criterion.
  • animal cell growth promoter oil / fat composition of the present invention By using the animal cell growth promoter oil / fat composition of the present invention to promote the growth of animal cells, it is possible to improve the productivity of animal cells or the production of useful bioactive substances produced by the cells. it can.
  • composition for external use on skin of the present invention has an ability to promote epithelial formation in humans and animals, and can be expected to have a healing effect in the treatment of stains and wrinkles, treatment of wounds (including pressure sores), etc. due to metabolism of the epidermis. It is.

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Abstract

La présente invention se rapporte à une composition de graisse peu onéreuse, qui présente des propriétés très stables et est dotée d'une fonction favorisant la croissance de cellules animales, à un milieu pour la culture de cellules animales contenant cette composition de graisse, et à une préparation cutanée externe ayant pour effet d'améliorer ou de soigner des symptômes cutanés. L'invention se rapporte plus précisément à une composition de graisse permettant de favoriser la croissance de cellules animales qui est une composition de graisse comportant un triglycéride, un diglycéride, un monoglycéride et un acide gras libre contenant au moins un élément sélectionné parmi de l'acide η-linoléique et un glycéride d'acide η-linoléique. L'invention se rapporte en outre à une préparation cutanée externe et à un milieu permettant de cultiver des cellules animales et contenant cette composition de graisse.
PCT/JP2004/018575 2003-12-17 2004-12-13 Composition de graisse favorisant la croissance de cellules animales, milieu de culture de cellules animales contenant ladite composition et preparation cutanee externe WO2005059114A1 (fr)

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JP2003419616A JP2005179211A (ja) 2003-12-17 2003-12-17 皮膚外用剤組成物
JP2003419615 2003-12-17
JP2003-419615 2003-12-17
JP2003-419616 2003-12-17

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3216858A4 (fr) * 2014-11-06 2018-07-11 Nippon Menard Cosmetic Co., Ltd. Agent destiné à maintenir des cellules souches à l'état indifférencié et agent destiné à favoriser leur croissance
JP2020002053A (ja) * 2018-06-28 2020-01-09 日本メナード化粧品株式会社 コラーゲン産生促進剤、ｍｍｐ阻害剤、メラニン生成抑制剤、細胞増殖促進剤、抗酸化剤、シワ改善剤、医薬品又は食品組成物

Citations (3)

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Publication number Priority date Publication date Assignee Title
JPS6314695A (ja) * 1986-07-08 1988-01-21 Suntory Ltd γ―リノレン酸及びこれを含有する脂質の製造方法
JPH1192343A (ja) * 1997-09-22 1999-04-06 Shiseido Co Ltd 毛髪成長期延長剤
JP2001163761A (ja) * 1999-12-03 2001-06-19 Koowa Techno Search:Kk ケミカルピーリング用化粧料

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Publication number Priority date Publication date Assignee Title
JPS6314695A (ja) * 1986-07-08 1988-01-21 Suntory Ltd γ―リノレン酸及びこれを含有する脂質の製造方法
JPH1192343A (ja) * 1997-09-22 1999-04-06 Shiseido Co Ltd 毛髪成長期延長剤
JP2001163761A (ja) * 1999-12-03 2001-06-19 Koowa Techno Search:Kk ケミカルピーリング用化粧料

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HARBIGEL L.S. ET AL: "Prevention of experimental autoimmune encephalomyelitis in Lewis rats by a novel fungal source of y-linolenic acid.", J. NUTR., vol. 74, no. 5, 1995, pages 701 - 715, XP009035788 *
TANAKA MINOHISA ET AL: "Y-linolenic acid o Keshohin ni Haigoshita Tokino Koka.", FRAG.J., vol. 30, no. 8, 2002, pages 86 - 88, XP002997487 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3216858A4 (fr) * 2014-11-06 2018-07-11 Nippon Menard Cosmetic Co., Ltd. Agent destiné à maintenir des cellules souches à l'état indifférencié et agent destiné à favoriser leur croissance
JP2020002053A (ja) * 2018-06-28 2020-01-09 日本メナード化粧品株式会社 コラーゲン産生促進剤、ｍｍｐ阻害剤、メラニン生成抑制剤、細胞増殖促進剤、抗酸化剤、シワ改善剤、医薬品又は食品組成物
JP7148115B2 (ja) 2018-06-28 2022-10-05 日本メナード化粧品株式会社 コラーゲン産生促進剤、ｍｍｐ阻害剤、メラニン生成抑制剤、細胞増殖促進剤、抗酸化剤、シワ改善剤、医薬品又は食品組成物

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