WO2013154407A1 - Anti-inflammatory, skin whitening and anti-aging composition comprising supernatant of lactic acid bacteria lysate - Google Patents

Anti-inflammatory, skin whitening and anti-aging composition comprising supernatant of lactic acid bacteria lysate Download PDF

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WO2013154407A1
WO2013154407A1 PCT/KR2013/003158 KR2013003158W WO2013154407A1 WO 2013154407 A1 WO2013154407 A1 WO 2013154407A1 KR 2013003158 W KR2013003158 W KR 2013003158W WO 2013154407 A1 WO2013154407 A1 WO 2013154407A1
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inflammatory
lactobacillus
composition
skin whitening
lactic acid
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PCT/KR2013/003158
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French (fr)
Korean (ko)
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정대균
정봉준
김혜림
유가은
정지혜
김태락
홍의범
최경숙
이종석
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주식회사 알엔에이
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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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • 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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9728—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • 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
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A61K8/365—Hydroxycarboxylic acids; Ketocarboxylic acids
    • 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
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof

Definitions

  • the present invention relates to a composition for anti-inflammatory, skin whitening and anti-aging comprising a supernatant of lactic acid bacteria lysate. More specifically, the present invention relates to a composition for anti-inflammatory, skin whitening and anti aging comprising a supernatant of lactic acid bacteria lysate having a size of 100 to 900 nm.
  • melanin The most important pigment that determines skin color is melanin, which is synthesized in the melanosomes in the melanocytes present in the basal layer of the epidermis. Melanin absorbs the light energy of sunlight UV to protect skin organs under the dermis from UV damage and protects the skin from external harmful factors such as trapping harmful oxygen and free radicals generated in skin Play a useful role.
  • the skin color of a person is determined by the amount of melanin in the skin cells. People with high melanin pigments have brown or black skin, while those with low melanin pigments have white skin.
  • Prostaglandin E2 is a metabolite in the arachidonic acid cascade, which promotes pain-inducing, pre- or anti-inflammatory activity, uterine contractile activity, and digestive system.
  • PGE2 is produced in keratinocytes by UV stimulation among the cells of the skin, and the generated PGE2 is released into the cells and acts as a lipid signaling intermadiate for immunoregulation, cell growth, cell differentiation, Affects carcinogenesis Therefore, inhibiting the production of PGE2 can obtain an anti-inflammatory effect.
  • PGE2 has been reported to stimulate melanocyte dendrite through PKC ⁇ in skin structures.
  • the melanocyte dendritic crystal is an important factor involved in the migration of the produced melanosomes to keratinocytes.
  • reducing PGE2 that stimulates melanocyte dendritic crystals inhibits pigmentation by UV-stimulated keratinocytes, resulting in a depigment effect.
  • Lactobacillus is a bacterium that plays a beneficial role in the human body and produces lactic acid (lactic acid) by fermenting sugars such as carbohydrates.
  • lactic acid bacteria Up to 400 kinds of lactic acid bacteria have been discovered so far, and 18 of them are known to be commercially available.
  • These lactic acid bacteria are known to have a physiological activity of immune enhancement, in particular, gastrointestinal immunity enhancement, and the consumption of fermented milk products is enormous.
  • various cosmetic materials that are beneficial to the body are developed. Although the effect of reducing wrinkles has been reported, in many cases, the effect received when using cosmetics was insignificant.
  • the present invention has been found to have anti-inflammatory, skin whitening and anti-aging effects according to the particle size of the lactic acid bacteria lysate.
  • the present invention relates to a composition for anti-inflammatory, skin whitening and anti aging comprising a supernatant of lactic acid bacteria lysate having a size of 100 to 900 nm.
  • the present invention provides a composition for anti-inflammatory, skin whitening and anti-aging comprising a supernatant of lactic acid bacteria lysate.
  • the present invention also provides an anti-inflammatory, skin whitening and anti-aging composition for use as an external preparation for skin, cosmetics and health food.
  • Lactic acid bacteria lysate in the present invention can increase the activity of the existing lactic acid bacteria through the atomization, and has the effect of inhibiting anti-inflammatory, inhibition of melanin synthesis and photo aging can be used as a cosmetic, pharmaceutical and health food.
  • 1 to 3 are graphs showing the inhibitory ability of the transient immune response in immune cells according to the particle size of Lactobacillus plantarium K8, Lactobacillus sakei, and Lactobacillus rhamnosus debris, respectively.
  • 4 to 6 are graphs showing the inhibitory ability of the transient immune response in intestinal cells according to the particle size of Lactobacillus plantarium K8, Lactobacillus sakei, and Lactobacillus rhamnosus debris, respectively.
  • Figure 7 is a graph of the inhibition of melanin production according to the particle size of the lactic acid bacteria lysate.
  • the present invention provides a composition for anti-inflammatory, skin whitening and anti-aging comprising a water-soluble active ingredient through lactic acid bacteria disruption and extraction.
  • Lactobacillus crushing and extraction of the present invention is to extract the supernatant from which the precipitate is removed by centrifugation after crushing the lactic acid bacteria, but not limited to the crushing method, heat killed, crushed using a high pressure homogenizer (Microfludizer) can do. More specifically, it can be crushed at a pressure of 10000 to 30000psi using a high pressure homogenizer (Microfludizer).
  • the lactic acid bacteria supernatant may be a water-soluble active ingredient obtained through crushing of lactic acid bacteria.
  • the molecular weight of the water-soluble active ingredient may be 391 to 597 m / z, and may include LTA and / or fatty acid.
  • the lactic acid bacteria may be Lactobacillus, Bifidobacterium, Streptococcus or Lactococcus.
  • Lactobacillus Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus rhamnosus, Lactobacillus sakei, Lactobacillus delbrueckii, Lactobacillus delbrueckii, Lactobacillus exoculus , Lactobacillus gasseri, Lactoacillus johnsonni, Lactobacillus helveticus or Lactobacillus casi.
  • the Bifidobacterium can be Bifidobacterium bifido, Bifidobacterium longum, Bifidobacterium infantis or Bifidobacterium animalis, or bifidobacterium animalis.
  • the Streptococcus may be Streptococcus thermophilus, and the Lactococcus may be Lactococcus lactis.
  • the size of the lactic acid bacteria lysate may be 100 to 900 nm, 300 to 600 nm, it can be seen that the anti-inflammatory, skin whitening and anti-aging effect is high when the size (see FIGS. 1 to 8).
  • the present invention also provides an anti-inflammatory, skin whitening and anti-aging composition for use as an external preparation for skin.
  • composition of the present invention may further include a known active substance as long as it does not have a deleterious effect on anti-inflammatory, skin whitening and anti-aging effects other than the supernatant of the lactic acid bacteria lysate.
  • the formulation of the external preparation for skin is not particularly limited.
  • formulations such as powder, gel, ointment, cream, liquid, aerosol and the like.
  • composition for anti-inflammatory, skin whitening and anti-aging comprising a supernatant of lactic acid bacteria lysate according to the present invention can be used as a cosmetic.
  • the cosmetics in addition to the supernatant of the lactic acid bacteria lysate according to the present invention may be added a material selected arbitrarily according to the formulation or purpose of use.
  • a material selected arbitrarily according to the formulation or purpose of use For example, purified water, oils, surfactants, moisturizers, higher alcohols, thickeners, chelating agents, pigments, fatty acids, antioxidants, preservatives, waxes, pH adjusters, fragrances and the like can be added.
  • the oils include mineral oil, cyclomethicone, squalane, octyldodecyl myristate, olive oil, macadamia nut oil, glycerol octanoate, castor oil, ethylhexyl isononanoate, dimethicone, cyclopentasiloxane, Sunflower seed oil and the like can be used.
  • the surfactants include sorbitan sesquinolate, polysorbate 60, glyceryl stearate, lipophilic glyceryl stearate, sorbitan oleate, sorbitan stearate, die-cetyl phosphate, sorbitan stearate / Sucrose cocoate, glyceryl stearate / polyethylene glycol-100 stearate, ceteareth-6 oleate, arachidyl alcohol / behenyl alcohol / arachidyl glucoside.
  • Polypropylene glycol-26-butes-26 / polyethylene glycol-40 hydrogenated castor oil and the like can be used.
  • glycerin As the moisturizing agent, glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycerin-26, methylgluse-20 and the like may be used. Cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol and the like may be used as the higher alcohol.
  • Xanthan gum carbomer, magnesium aluminum silicate, cellulose gum, dextrin palmitate may be used as the thickener.
  • disodium idieti As the chelating agent, disodium idieti, tetrasodium idieti and the like may be used.
  • dye Blue No. 1, Red No. 40, Yellow No. 4, Yellow No. 5, etc. may be used.
  • glycerin As the moisturizing agent, glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycerin-26, methylgluse-20 and the like may be used.
  • Cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol and the like may be used as the higher alcohol.
  • Xanthan gum carbomer, magnesium aluminum silicate, cellulose gum, dextrin palmitate may be used as the thickener.
  • disodium idieti As the chelating agent, disodium idieti, tetrasodium idieti and the like may be used.
  • dye Blue No. 1, Red No. 40, Yellow No. 4, Yellow No. 5, etc. may be used.
  • fatty acid stearic acid, mystic acid, palmitic acid, isostearic acid, lauric acid, or the like may be used.
  • Tocopheryl acetate, butylated hydroxytoluene, etc. may be used as the antioxidant.
  • methyl paraben, butyl paraben, propyl paraben, phenoxyethanol, imidazolidinyl urea, chlorphenesin and the like may be used.
  • Carnauba wax, candenilla wax, beeswax, braze and the like may be used as the wax.
  • pH adjusting agent triethanolamine, citric acid, or the like may be used.
  • fragrance natural fragrance, combination fragrance, or the like may be used.
  • the formulation of the cosmetic prepared with the composition of the present invention is not particularly limited, and may have, for example, a formulation such as a nourishing cream, astringent lotion, a flexible lotion, a lotion, an essence, a nourishing gel or a massage cream.
  • the present invention also provides a composition for anti-inflammatory, skin whitening and anti-aging for use as a health food.
  • composition for anti-inflammatory, skin whitening and anti-aging including the supernatant of the lactic acid bacteria lysate according to the present invention can be added, for example, various foods, beverages, gums, teas, vitamin complexes, health functional foods, etc. There is this.
  • the amount of the anti-inflammatory, skin whitening and anti-aging composition in the health food may be added in 0.01 to 15% by weight of the total food weight, the amount of the anti-inflammatory, skin whitening and anti-aging composition in the beverage may be from 0.02 to 100 ml It can be added at a ratio of 5 g, 0.3 to 1 g.
  • composition for anti-inflammatory, skin whitening and anti-aging comprising the supernatant of the lactic acid bacteria lysate according to the present invention
  • natural carbohydrates examples include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents tacumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
  • the proportion of natural carbohydrates is generally about 1-20 g, about 5-12 g per 100 ml of the composition of the present invention.
  • composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
  • compositions of the present invention may include a pulp for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but it is generally chosen that the composition of the invention is selected in the range of 0 to about 20 parts by weight per 100 parts by weight.
  • Lactobacillus plantarium K8, Lactobacillus sakei, and Lactobacillus rhamnosus at 1 ⁇ 10 10 10 cells / ml were prepared, and heat killed at 80 ° C., and disrupted using a Microfludizer.
  • the size of the crushed sample was 100-900 nm (using RNA holding strain).
  • Monocyte THP-1 cells were inoculated (seeding) at 2 ⁇ 10 5 cells / ml, stabilized for 4 hours, and then treated with crushed light samples and incubated for 24 hours. Thereafter, the mixture was treated with LPS (500 ng / ml) for 4 hours, and confirmed by sandwich ELISA.
  • TNF-alpha was inhibited by the lactic acid bacteria water-soluble active ingredient. This means that it has an immunomodulatory function in vivo through inhibition of transient immune response.
  • HT-29 cells colon epithelial cells, were seeded at 2 ⁇ 10 5 cells / ml, stabilized for 12 hours, and pre-treated with crushed light samples (24h), and then incubated with S.aureus for 6 hours. , sandwich ELISA was confirmed.
  • Human normal dermal Fibroblast cells were inoculated at 2 ⁇ 10 5 cells / ml, stabilized for 12 hours, and pre-treated with crushed light samples (24h) and then changed to serum free media. After UV 25mJ / cm 2 treatment, it was incubated for 24 hours, and confirmed by Sandwich ELISA.
  • the degree of photoaging inhibition was confirmed through the decrease of the expression of MMP-1 increased by UV. Increasing the skin aging inhibition by the lactic acid bacteria water-soluble active ingredient produced by the crushing was able to increase the active material induced by the degree of crushing and increase the skin aging inhibitory activity of the lactic acid bacteria.

Abstract

The present invention relates to an anti-inflammatory, skin whitening and anti-aging composition comprising supernatant of lactic acid bacteria lysate. More particularly, the present invention relates to an anti-inflammatory, skin whitening and anti-aging composition comprising supernatant of lactic acid bacteria lysate having a size of 100 to 900nm. The lactic acid bacteria lysate of the present invention may increase the activity of existing lactic acid bacteria through atomization, and has the effect of controlling inflammation, inhibiting melanine synthesis and inhibiting photoaging, and therefore, can be used as a cosmetic, a pharmaceutical agent and a health food.

Description

유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물Anti-inflammatory, skin whitening and anti-aging composition comprising supernatant of lactic acid bacteria lysate
본 발명은 유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물에 관한 것이다. 보다 구체적으로 본 발명은 100 내지 900nm의 크기를 갖는 유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물에 관한 것이다.The present invention relates to a composition for anti-inflammatory, skin whitening and anti-aging comprising a supernatant of lactic acid bacteria lysate. More specifically, the present invention relates to a composition for anti-inflammatory, skin whitening and anti aging comprising a supernatant of lactic acid bacteria lysate having a size of 100 to 900 nm.
건강하고 여유로운 삶이 요구되고 있는 가운데 아름다워지고자 하는 소망과 함께 기능성 화장품에 대한 관심이 더욱 높아지고 있고, 그에 따라서 기능성 화장품은 단순히 미용의 개념에서 진화하여 고기능, 다기능의 개념을 포함한 미백 등의 기능성 화장품의 신소재 원료 개발이 강하게 요구되고 있다. 앞으로의 기능성 화장품이 확대됨에 따라 미백 제품은 기능성 화장품의 30% 이상을 차지할 것이라 예상하며 기능성 화장품 개발에 있어서 중추적인 역할을 할 것이라 예측된다.Along with the desire to be beautiful with a healthy and leisurely life, interest in functional cosmetics is increasing. Accordingly, functional cosmetics have simply evolved from the concept of beauty, and the functional cosmetics, such as whitening including high functional and multifunctional concepts, There is a strong demand for the development of new materials. As functional cosmetics expand in the future, whitening products are expected to occupy more than 30% of functional cosmetics and are expected to play a pivotal role in the development of functional cosmetics.
피부색을 결정하는 가장 중요한 색소는 멜라닌으로서 표피의 기저층에 존재하는 멜라노사이트(melanocyte) 내의 멜라노좀(melanosome)에서 합성된다. 멜라닌은 햇빛 UV의 빛 에너지를 흡수하여 UV에 의한 손상으로부터 진피 이하의 피부 기관을 보호하는 역할을 하며, 피부 생체 내에 생겨난 유해산소 및 프리라디칼 등을 잡아주는 등 외부 유해인자로부터 피부를 보호해 주는 유용한 역할을 수행한다. 또한, 피부 세포 내의 멜라닌 양에 따라 사람의 피부색을 결정한다. 멜라닌 색소가 많은 사람은 갈색 또는 검은 색 피부를 갖는 반면, 멜라닌 색소가 적은 사람은 흰색의 피부를 갖게 된다. 하지만 일광, 호르몬변화, 염증 또는 약제 등에 의해 멜라닌이 과도로 합성되거나 각화작용이 원활히 이루어지지 않아 색소가 침착되면 기미, 주근깨와 같은 피부질병을 일으키며 또한 피부암의 원인이 되기도 한다. 따라서, 미용욕구 충족뿐만 아니라 피부질환 치료를 위한 미백제에 대한 개발이 필요하다.The most important pigment that determines skin color is melanin, which is synthesized in the melanosomes in the melanocytes present in the basal layer of the epidermis. Melanin absorbs the light energy of sunlight UV to protect skin organs under the dermis from UV damage and protects the skin from external harmful factors such as trapping harmful oxygen and free radicals generated in skin Play a useful role. In addition, the skin color of a person is determined by the amount of melanin in the skin cells. People with high melanin pigments have brown or black skin, while those with low melanin pigments have white skin. However, due to daylight, hormonal changes, inflammation, or drugs, too much melanin synthesis or keratinization does not work well, and pigment deposits can cause skin diseases such as blemishes and freckles, and can also cause skin cancer. Therefore, it is necessary to develop a whitening agent for treating skin diseases as well as satisfying beauty needs.
멜라닌의 생성을 억제하는 다양한 접근방법이 있는데, 예를 들면 티로시네즈(tyrosinase) 생성 억제, 티로시네즈 활성 억제, 기존 멜라닌의 환원, 광산화 억제 또는 멜라닌 배설 촉진 등이 있다.There are various approaches to inhibit the production of melanin, for example, inhibiting tyrosinase production, inhibiting tyrosinase activity, reducing existing melanin, inhibiting photooxidation or promoting melanin excretion.
프로스타글란딘 E2(prostaglandin E2, PGE2)는 아라키돈산 반응단계(arachidonate cascade)에 있어서의 대사 산물의 하나로서 활성을 유도하는 통증, 전- 또는 항-염증성 활성, 자궁수축 활성, 소화 연동에 대한 촉진Prostaglandin E2 (PGE2) is a metabolite in the arachidonic acid cascade, which promotes pain-inducing, pre- or anti-inflammatory activity, uterine contractile activity, and digestive system.
작용, 각성 활성, 위산분비에 대한 억제 작용, 저혈압 활성, 혈소판 저해 활성, 골-재흡수 활성 및 혈관 형성활성 등의 각종 활성을 지닌다.It has various activities such as action, arousal activity, inhibitory action against gastric acid secretion, hypotension activity, platelet inhibitory activity, bone-resorption activity and angiogenesis activity.
PGE2는 피부의 세포 중에서는 UV 자극에 의해 케라티노사이트(keratinocyte)에서 생성되며, 상기 생성된 PGE2는 세포 외로 방출되어 지질 신호 매개체(lipid signaling intermadiate)로 작용하여 면역조절, 세포 성장, 세포분화 및 발암(carcinogenesis)에 영향을 미친다. 따라서 PGE2의 생성을 억제하면 항염 효과를 얻을 수 있다.PGE2 is produced in keratinocytes by UV stimulation among the cells of the skin, and the generated PGE2 is released into the cells and acts as a lipid signaling intermadiate for immunoregulation, cell growth, cell differentiation, Affects carcinogenesis Therefore, inhibiting the production of PGE2 can obtain an anti-inflammatory effect.
PGE2는 피부 구조에서 PKCζ을 통하여 멜라닌세포 수지상 결정(melanocyte dendrite)을 자극하는 결과가 보고된바 있다. 상기 멜라닌세포 수지상 결정은 생성된 멜라노좀(melanosome)이 케라티노사이트로 이동되는데 관여하는 중요한 요소이다. 즉, 멜라닌세포 수지상 결정을 자극시키는 PGE2를 감소시키는 것은 UV에 의해 자극된 케라티노사이트가 색소형성(pigmentation)을 일으키는 것을 억제하여, 탈색소(depigment) 효과가 있다.PGE2 has been reported to stimulate melanocyte dendrite through PKCζ in skin structures. The melanocyte dendritic crystal is an important factor involved in the migration of the produced melanosomes to keratinocytes. In other words, reducing PGE2 that stimulates melanocyte dendritic crystals inhibits pigmentation by UV-stimulated keratinocytes, resulting in a depigment effect.
유산균은 인체 내에서 유익한 역할을 하는 세균으로서 탄수화물 같은 당을 발효시켜 유산(젖산)을 생성하는 균으로, 지금까지 400여종이 발견되었으며, 그 중 상품성이 있는 것은 18종으로 알려져 있다. 이들 유산균은 면역 증강 특히, 위 장관 면역 증강의 생리활성을 가진다고 알려져 있어, 발효유제품의 소비량은 실로 어마어마하다. 특히, 최근 웰빙 트렌드에 따라, 먹어서 좋은 식품이 발랐을 때도 좋을 것이라는 사회적 분위기가 조성됨에 따라, 몸에 이로운 다양한 화장품 소재들이 개발되면서 유산균의 배양액이 피부의 보습, 각질층의 턴오버(turn over)속도 증가, 주름살 완화 등의 효과가 보고되어 있으나, 실제로 화장품을 사용하였을 때 전달받는 그 효과가 미미한 경우가 많았다.Lactobacillus is a bacterium that plays a beneficial role in the human body and produces lactic acid (lactic acid) by fermenting sugars such as carbohydrates. Up to 400 kinds of lactic acid bacteria have been discovered so far, and 18 of them are known to be commercially available. These lactic acid bacteria are known to have a physiological activity of immune enhancement, in particular, gastrointestinal immunity enhancement, and the consumption of fermented milk products is enormous. In particular, according to the recent well-being trend, as a social atmosphere that is good to eat good food is created, various cosmetic materials that are beneficial to the body are developed. Although the effect of reducing wrinkles has been reported, in many cases, the effect received when using cosmetics was insignificant.
또한, 종래에는 유산균을 발효 배양하여 생성되는 산물을 화장료 조성물에 포함시키거나, 버섯, 쌀, 과즙, 식물 추출물 등을 특정 속의 유산균으로 발효시켜 얻어진 발효액을 화장료에 배합하여 사용하는 경우 등이 있었다. 하지만, 유산균의 발효액, 배양액이 아닌 유산균 자체의 파쇄물의 입자 크기에 따른 항 염증 및 피부노화 억제효과에 대해서는 아직까지 알려진 바가 없었다.In addition, conventionally, products produced by fermenting and lactic acid bacteria were included in the cosmetic composition, or a fermentation broth obtained by fermenting mushrooms, rice, fruit juice, plant extracts, and the like with a specific genus was used in a cosmetic composition. However, the anti-inflammatory and skin aging inhibitory effect according to the particle size of the fermentation broth of the lactic acid bacteria, the lysate of the lactic acid bacteria itself, not the culture medium has not been known so far.
이에 본 발명은 유산균 파쇄물의 입자 크기에 따른 항 염증, 피부 미백 및 항 노화 효능이 있음을 발견하게 되었다.Accordingly, the present invention has been found to have anti-inflammatory, skin whitening and anti-aging effects according to the particle size of the lactic acid bacteria lysate.
따라서, 본 발명은 100 내지 900nm의 크기를 갖는 유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물에 관한 것이다.Accordingly, the present invention relates to a composition for anti-inflammatory, skin whitening and anti aging comprising a supernatant of lactic acid bacteria lysate having a size of 100 to 900 nm.
본 발명은 유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물을 제공한다. The present invention provides a composition for anti-inflammatory, skin whitening and anti-aging comprising a supernatant of lactic acid bacteria lysate.
또한, 본 발명은 피부 외용제, 화장료 및 건강식품으로 사용하는 항 염증, 피부 미백 및 항 노화용 조성물을 제공한다.The present invention also provides an anti-inflammatory, skin whitening and anti-aging composition for use as an external preparation for skin, cosmetics and health food.
본 발명에 유산균 파쇄물은 기존 유산균이 갖는 활성을 미립화를 통해 증대 시킬 수 있고, 항 염증, 멜라닌 합성의 저해 및 광 노화를 억제하는 효과가 있어 화장료, 약제 및 건강식품으로 이용될 수 있다.Lactic acid bacteria lysate in the present invention can increase the activity of the existing lactic acid bacteria through the atomization, and has the effect of inhibiting anti-inflammatory, inhibition of melanin synthesis and photo aging can be used as a cosmetic, pharmaceutical and health food.
도 1 내지 도 3은 각각 락토바실러스 플랜타륨 K8, 락토바실러스 사케이, 락토바실러스 람노서스 파쇄물의 입자 크기에 따른 면역세포에서의 과도면역반응의 억제능에 관한 그래프이다.1 to 3 are graphs showing the inhibitory ability of the transient immune response in immune cells according to the particle size of Lactobacillus plantarium K8, Lactobacillus sakei, and Lactobacillus rhamnosus debris, respectively.
도 4 내지 도 6은 각각 락토바실러스 플랜타륨 K8, 락토바실러스 사케이, 락토바실러스 람노서스 파쇄물의 입자 크기에 따른 장 세포에서의 과도면역반응의 억제능에 관한 그래프이다.  4 to 6 are graphs showing the inhibitory ability of the transient immune response in intestinal cells according to the particle size of Lactobacillus plantarium K8, Lactobacillus sakei, and Lactobacillus rhamnosus debris, respectively.
도 7은 유산균 파쇄물의 입자 크기에 따른 멜라닌 생성의 억제능에 관한 그래프이다.Figure 7 is a graph of the inhibition of melanin production according to the particle size of the lactic acid bacteria lysate.
도 8은 유산균 파쇄물의 입자 크기에 따른 MMP-1의 억제능에 관한 그래프이다.8 is a graph showing the inhibitory ability of MMP-1 according to the particle size of the lactic acid bacteria lysate.
본 발명은 유산균 파쇄 및 추출을 통한 수용성 활성성분을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물을 제공한다.The present invention provides a composition for anti-inflammatory, skin whitening and anti-aging comprising a water-soluble active ingredient through lactic acid bacteria disruption and extraction.
본 발명의 유산균 파쇄 및 추출이란 유산균을 파쇄한 후 원심분리를 통해 침전물을 제거한 상등액을 추출하는 것으로서, 파쇄방법으로는 이에 제한되는 것은 아니나, Heat killed, 고압호모게나이저(Microfludizer)를 이용하여 파쇄할 수 있다. 보다 구체적으로 고압호모게나이저(Microfludizer)를 이용하여 10000 내지 30000psi의 압력으로 파쇄할 수 있다.Lactobacillus crushing and extraction of the present invention is to extract the supernatant from which the precipitate is removed by centrifugation after crushing the lactic acid bacteria, but not limited to the crushing method, heat killed, crushed using a high pressure homogenizer (Microfludizer) can do. More specifically, it can be crushed at a pressure of 10000 to 30000psi using a high pressure homogenizer (Microfludizer).
상기 유산균 상등액은 유산균의 파쇄를 통해 얻은 수용성 활성성분일 수 있다.The lactic acid bacteria supernatant may be a water-soluble active ingredient obtained through crushing of lactic acid bacteria.
상기 수용성 활성성분의 분자량은 391 내지 597 m/z일 수 있고, LTA 및/또는 fatty acid를 포함할 수 있다.The molecular weight of the water-soluble active ingredient may be 391 to 597 m / z, and may include LTA and / or fatty acid.
상기 유산균은 락토바실러스, 비피도박테리움, 스트렙토코커스 또는 락토코커스일 수 있다. The lactic acid bacteria may be Lactobacillus, Bifidobacterium, Streptococcus or Lactococcus.
상기 락토바실러스는 락토바실러스 플랜타륨(Lactobacillus plantarum), 락토바실러스 람노서스(Lactobacillus rhamnosus), 락토바실러스 사케이(lactobacillus sakei), 락토바실러스 델브르키(Lactobacillus delbrueckii), 락토바실러스 엑시도필러스(Lactobacillus acidophilus), 락토바실러스 가세리 (Lactobacillus gasseri), 락토바실러스 존스니(Lactoacillus johnsonni), 락토바실러스 헬베티쿠스(Lactobacillus helveticus) 또는 락토바실러스 카세이(Lactobacillus casei)일 수 있다. The Lactobacillus Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus rhamnosus, Lactobacillus sakei, Lactobacillus delbrueckii, Lactobacillus delbrueckii, Lactobacillus exoculus , Lactobacillus gasseri, Lactoacillus johnsonni, Lactobacillus helveticus or Lactobacillus casi.
상기 비피도박테리움은 비피도박테리움 비피도(Bifidobacterium bifido), 비피도박테리움 롱검(Bifidobacterium longum), 비피도박테리움 인판티스(Bifidobacterium infantis) 또는 비피도박테리움 아니말리스(bifidobacterium animalis)일 수 있으며, 상기 스트렙토코커스는 스트렙토코커스 써모필러스(Streptococcus thermophilus)일 수 있으며, 상기 락토코커스는 락토코커스 락티스 (Lactococcus lactis)일 수 있다.The Bifidobacterium can be Bifidobacterium bifido, Bifidobacterium longum, Bifidobacterium infantis or Bifidobacterium animalis, or bifidobacterium animalis. The Streptococcus may be Streptococcus thermophilus, and the Lactococcus may be Lactococcus lactis.
본 발명의 실시예에서 유산균 파쇄물의 크기는 100 내지 900nm, 300 내지 600nm일 수 있고, 상기 크기를 갖는 경우 항 염증, 피부 미백 및 항 노화 효과가 높음을 알 수 있었다(도 1 내지 8참조). In the embodiment of the present invention, the size of the lactic acid bacteria lysate may be 100 to 900 nm, 300 to 600 nm, it can be seen that the anti-inflammatory, skin whitening and anti-aging effect is high when the size (see FIGS. 1 to 8).
또한, 본 발명은 피부 외용제로 사용하는 항 염증, 피부 미백 및 항 노화용 조성물을 제공한다.The present invention also provides an anti-inflammatory, skin whitening and anti-aging composition for use as an external preparation for skin.
본 발명의 조성물은 상기 유산균 파쇄물의 상등액 이외 항 염증, 피부 미백 및 항 노화 효과에 유해한 효과를 끼치지 않는 한, 공지된 활성 물질을 추가로 포함할 수 있다.The composition of the present invention may further include a known active substance as long as it does not have a deleterious effect on anti-inflammatory, skin whitening and anti-aging effects other than the supernatant of the lactic acid bacteria lysate.
상기 피부 외용제의 제형은 특별히 제한되지 않는다. 예를 들어, 파우더(power), 젤(gel), 연고(ointment), 크림(cream), 액체(liquid), 에어로졸(aerosol) 등의 제형을 가진다.The formulation of the external preparation for skin is not particularly limited. For example, there are formulations such as powder, gel, ointment, cream, liquid, aerosol and the like.
또한, 본 발명에 따라 유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물은 화장료로 사용할 수 있다. 상기 화장료로 사용시 본 발명에 따른 유산균 파쇄물의 상등액 외에 기타 화장료의 제형 또는 사용목적 등에 맞게 임의로 선정한 물질을 첨가할 수 있다. 예를 들어, 정제수, 유분, 계면활성제, 보습제, 고급 알코올, 증점제, 킬레이트제, 색소, 지방산, 산화방지제, 방부제, 왁스, pH 조절제, 향료 등이 첨가될 수 있다.In addition, the composition for anti-inflammatory, skin whitening and anti-aging comprising a supernatant of lactic acid bacteria lysate according to the present invention can be used as a cosmetic. When used as the cosmetics, in addition to the supernatant of the lactic acid bacteria lysate according to the present invention may be added a material selected arbitrarily according to the formulation or purpose of use. For example, purified water, oils, surfactants, moisturizers, higher alcohols, thickeners, chelating agents, pigments, fatty acids, antioxidants, preservatives, waxes, pH adjusters, fragrances and the like can be added.
상기 유분으로는 미네랄오일, 사이클로메치콘, 스쿠알란, 옥틸도데실 미리스테이트, 올리브오일, 마카다미아 너트오일,　글리세릴옥타노에이트, 캐스터오일, 에칠헥실 이소노나노에이트, 디메치콘, 사이크로펜타실록산, 선플라워씨드오일 등이 사용될 수 있다.　The oils include mineral oil, cyclomethicone, squalane, octyldodecyl myristate, olive oil, macadamia nut oil, glycerol octanoate, castor oil, ethylhexyl isononanoate, dimethicone, cyclopentasiloxane, Sunflower seed oil and the like can be used.
상기 계면활성제로는 소르비탄세스퀴놀리에이트, 폴리솔베이트 60, 글리세릴 스테아레이트, 친유형 글리세릴스테아레이트, 소르비탄 올리에이트, 소르비탄 스테아레이트, 디이에이-세틸포스페이트, 소르비탄스테아레이트/ 슈크로스코코에이트, 글리세릴스테아레이트/ 폴리에틸렌글라이콜-100 스테아레이트, 세테아레스-6 올리베이트, 아라키딜알코올/베헤닐알코올/아라키딜 글루코사이드. 폴리프로필렌글라이콜-26-부테스-26/폴리에틸렌글라이콜-40 하이드로제네이티드 캐스터오일 등이 사용될 수 있다.　The surfactants include sorbitan sesquinolate, polysorbate 60, glyceryl stearate, lipophilic glyceryl stearate, sorbitan oleate, sorbitan stearate, die-cetyl phosphate, sorbitan stearate / Sucrose cocoate, glyceryl stearate / polyethylene glycol-100 stearate, ceteareth-6 oleate, arachidyl alcohol / behenyl alcohol / arachidyl glucoside. Polypropylene glycol-26-butes-26 / polyethylene glycol-40 hydrogenated castor oil and the like can be used.
상기 보습제로는 글리세린, 솔비톨, 프로필렌 글라이콜, 부틸렌 글라이콜, 헥실렌 글라이콜, 디글리세린, 베타인, 글리세레스-26, 메칠글루세스-20 등이 사용될 수 있다.　 상기 고급 알코올로는 세틸알코올, 스테아릴 알코올, 옥틸도데칸올, 이소스테아릴 알코올 등이 사용될 수 있다.As the moisturizing agent, glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycerin-26, methylgluse-20 and the like may be used. Cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol and the like may be used as the higher alcohol.
상기 증점제로는 산탄검, 카보머, 마그네슘알루미늄실리케이트, 셀룰로오스검, 덱스트린 팔미테이트 등이 사용될 수 있다.Xanthan gum, carbomer, magnesium aluminum silicate, cellulose gum, dextrin palmitate may be used as the thickener.
상기 킬레이트제로는 디소듐이디티에이, 테트라소듐이디티에이 등이 사용될 수 있다. 상기 색소로는 청색1호, 적색40호, 황색4호, 황색 5호 등이 사용될 수 있다.　As the chelating agent, disodium idieti, tetrasodium idieti and the like may be used. As the dye, Blue No. 1, Red No. 40, Yellow No. 4, Yellow No. 5, etc. may be used.
상기 보습제로는 글리세린, 솔비톨, 프로필렌 글라이콜, 부틸렌 글라이콜, 헥실렌 글라이콜, 디글리세린, 베타인, 글리세레스-26, 메칠글루세스-20 등이 사용될 수 있다.　As the moisturizing agent, glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, betaine, glycerin-26, methylgluse-20 and the like may be used.
상기 고급 알코올로는 세틸알코올, 스테아릴 알코올, 옥틸도데칸올, 이소스테아릴 알코올 등이 사용될 수 있다.Cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol and the like may be used as the higher alcohol.
상기 증점제로는 산탄검, 카보머, 마그네슘알루미늄실리케이트, 셀룰로오스검, 덱스트린 팔미테이트 등이 사용될 수 있다.Xanthan gum, carbomer, magnesium aluminum silicate, cellulose gum, dextrin palmitate may be used as the thickener.
상기 킬레이트제로는 디소듐이디티에이, 테트라소듐이디티에이 등이 사용될 수 있다. 상기 색소로는 청색1호, 적색40호, 황색4호, 황색 5호 등이 사용될 수 있다.　As the chelating agent, disodium idieti, tetrasodium idieti and the like may be used. As the dye, Blue No. 1, Red No. 40, Yellow No. 4, Yellow No. 5, etc. may be used.
상기 지방산으로는 스테아릭애씨드, 미리스틱애씨드, 팔미틱애씨드, 이소스테아릭애씨드, 라우릭애씨드 등이 사용될 수 있다.As the fatty acid, stearic acid, mystic acid, palmitic acid, isostearic acid, lauric acid, or the like may be used.
상기 산화방지제로는 토코페릴아세테이드, 부틸레이티드하이드록시톨루엔 등이 사용될 수 있다.Tocopheryl acetate, butylated hydroxytoluene, etc. may be used as the antioxidant.
상기 방부제로는 메칠파라벤, 부틸파라벤, 프로필파라벤, 페녹시에탄올, 이미다졸리디닐우레아, 클로르페네신 등이 사용될 수 있다.As the preservative, methyl paraben, butyl paraben, propyl paraben, phenoxyethanol, imidazolidinyl urea, chlorphenesin and the like may be used.
상기 왁스로는 카나우바왁스, 칸데닐라왁스, 밀납, 경납 등이 사용될 수 있다.Carnauba wax, candenilla wax, beeswax, braze and the like may be used as the wax.
상기 pH조절제로는 트리에탄올아민, 씨트릭애씨드 등이 사용될 수 있다.As the pH adjusting agent, triethanolamine, citric acid, or the like may be used.
상기 향료로는 천연향료, 조합향료 등이 사용될 수 있다.As the fragrance, natural fragrance, combination fragrance, or the like may be used.
본 발명의 조성물로 제조되는 화장료의 제형은 특별히 제한되는 바가 없으며, 예를 들면, 영양 크림, 수렴 화장수, 유연 화장수, 로션, 에센스, 영양젤 또는 마사지 크림 등의 제형을 가질 수 있다.The formulation of the cosmetic prepared with the composition of the present invention is not particularly limited, and may have, for example, a formulation such as a nourishing cream, astringent lotion, a flexible lotion, a lotion, an essence, a nourishing gel or a massage cream.
또한, 본 발명은 건강식품으로 사용하는 항 염증, 피부 미백 및 항 노화 용 조성물을 제공한다.The present invention also provides a composition for anti-inflammatory, skin whitening and anti-aging for use as a health food.
본 발명에 따른 유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물을 첨가할 수 있는 건강식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강 기능성 식품류 등이 있다.As a health food to which the composition for anti-inflammatory, skin whitening and anti-aging including the supernatant of the lactic acid bacteria lysate according to the present invention can be added, for example, various foods, beverages, gums, teas, vitamin complexes, health functional foods, etc. There is this.
건강식품 중의 상기 항 염증, 피부 미백 및 항 노화용 조성물의 양은 전체 식품 중량의 0.01 내지 15 중량%로 가할 수 있으며, 음료 중의 상기 항 염증, 피부 미백 및 항 노화용 조성물의 양은 100 ㎖를 기준으로 0.02 내지 5 g, 0.3 내지 1 g의 비율로 가할 수 있다.The amount of the anti-inflammatory, skin whitening and anti-aging composition in the health food may be added in 0.01 to 15% by weight of the total food weight, the amount of the anti-inflammatory, skin whitening and anti-aging composition in the beverage may be from 0.02 to 100 ml It can be added at a ratio of 5 g, 0.3 to 1 g.
본 발명에 따른 유산균 파쇄물의 상등액을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물을 건강식품으로 사용할 때, 지시된 비율로 필수 성분으로서 상기 조성물을 포함하는 외에는 다른 성분에는 특별한 제한이 없으며, 통상의 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 포함할 수 있다.When using the composition for anti-inflammatory, skin whitening and anti-aging comprising the supernatant of the lactic acid bacteria lysate according to the present invention as a health food, there is no particular limitation to other ingredients except for including the composition as an essential ingredient in the indicated ratios, Various flavors or natural carbohydrates and the like may be included as additional ingredients.
상기 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제(사카린, 아스파르탐 등)을 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 1 내지 20 g, 약　5 내지 12 g이다.Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, about 5-12 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 포함할 수 있다.In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
그 밖에 본 발명의 조성물들은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 조성물을 100 중량부 당 0 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.In addition, the compositions of the present invention may include a pulp for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but it is generally chosen that the composition of the invention is selected in the range of 0 to about 20 parts by weight per 100 parts by weight.
이하 본 발명을 실시예에 의하여 더욱 상세하게 설명한다. 이들 실시예는 단지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시예에 국한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.
<실시예 1> 함 염증 효능Example 1 Inflammation Effect
1. 샘플 준비1. Sample Preparation
1x1010 cells/ml의 락토바실러스 플랜타륨 K8, 락토바실러스 사케이, 락토바실러스 람노서스를 준비, 80℃에서 Heat killed, Microfludizer를 이용하여 파쇄하였다. 파쇄된 샘플의 크기는 100~900 nm 였다(㈜ RNA 보유 균주를 이용함.)Lactobacillus plantarium K8, Lactobacillus sakei, and Lactobacillus rhamnosus at 1 × 10 10 cells / ml were prepared, and heat killed at 80 ° C., and disrupted using a Microfludizer. The size of the crushed sample was 100-900 nm (using RNA holding strain).
각 파쇄 샘플을 원심분리를 통해 침전물을 제거, 상등액을 수집하여 활성검사에 사용하였다. Each crushed sample was centrifuged to remove sediment, and the supernatant was collected and used for activity test.
2. 면역세포에서의 과도면역반응의 억제2. Inhibition of transient immune response in immune cells
Monocyte 인 THP-1 cell을 2×105cells/ml로 접종(seeding)을 하고, 4시간 안정화 후 파쇄상등 sample을 처리한 뒤 24시간 배양을 하였다. 그 후, LPS(500ng/ml)로 4 시간 동안 처리하고, sandwich ELISA로 확인하였다.Monocyte THP-1 cells were inoculated (seeding) at 2 × 10 5 cells / ml, stabilized for 4 hours, and then treated with crushed light samples and incubated for 24 hours. Thereafter, the mixture was treated with LPS (500 ng / ml) for 4 hours, and confirmed by sandwich ELISA.
유산균 수용성 활성성분에 의해 TNF-alpha의 과도한 발현이 억제됨을 확인할 수 있었다. 이는 과도면역반응의 억제를 통한 생체 내 면역조절기능을 가지고 있음을 의미한다. It was confirmed that excessive expression of TNF-alpha was inhibited by the lactic acid bacteria water-soluble active ingredient. This means that it has an immunomodulatory function in vivo through inhibition of transient immune response.
파쇄 크기의 감소에 따라 그러한 억제반응이 증가하다가, 100nm이하에서 억제활성이 사라지는 것을 확인할 수 있었다. 이는 유산균 파쇄 상등이 가지는 활성이 파쇄 정도에 따라 증가, 혹은 감소 할 수 있다고 말할 수 있었다. 파쇄 크기에 따라 활성이 증가, 감소할 수 있으며, 이는 여러 유산균에서 같은 경향을 보였다.It was confirmed that such inhibitory reaction increases with decreasing crushing size, and the inhibitory activity disappears below 100 nm. It could be said that the activity of lactic acid bacteria crushing supernatant may increase or decrease depending on the degree of crushing. Depending on the size of the crushing, the activity may increase or decrease, which shows the same tendency in various lactic acid bacteria.
3. 장 세포에서의 과도면역반응의 억제3. Inhibition of transient immune response in intestinal cells
Colon epithelial cell 인 HT-29 cell을 2×105cells/ml로 접종(seeding)을 하고, 12시간 안정화 후 파쇄상등 sample을 전처리(24h)한 뒤 배양한 후 S.aureus를 6시간 동안 처리하고, sandwich ELISA로 확인하였다.HT-29 cells, colon epithelial cells, were seeded at 2 × 10 5 cells / ml, stabilized for 12 hours, and pre-treated with crushed light samples (24h), and then incubated with S.aureus for 6 hours. , sandwich ELISA was confirmed.
과도한 장내 면역반응을 유도하고, 이를 통해 장내 과도 염증반응, 나아가 장 질환을 유도한다고 알려진 S.aureus에 의해 유도되는 장내 과도한 염증반응의 억제를 확인하였다. Excessive intestinal immune response was induced, which confirmed the suppression of excessive intestinal inflammatory response induced by S. aureus, which is known to induce intestinal transient inflammatory response and further bowel disease.
파쇄에 따른 크기 감소에 따른 활성의 증가(유도되는 과도한 염증반응의 감소정도)를 유산균 수용성 활성성분을 통해 확인할 수 있었다. 기존의 유산균이 가지는 활성이 파쇄를 통하여 증가될 수 있음을 확인하였다. 이는 기존 유산균이 가지는 면역활성의 증가를 가져올 수 있으며, 이를 통해, 더 높은 활성을 가지는 유산균제품의 생산을 유도할 수 있다(도 4 내지 6 참조).Increasing activity (decreased degree of excessive inflammatory response induced) according to the decrease in size due to crushing was confirmed by the lactic acid bacteria water-soluble active ingredient. It was confirmed that the activity of the existing lactic acid bacteria can be increased through crushing. This may lead to an increase in the immune activity of the existing lactic acid bacteria, through which it can induce the production of lactic acid bacteria products having a higher activity (see FIGS. 4 to 6).
<실시예 2> 피부 미백효과Example 2 Skin Whitening Effect
melan-a cell을 접종(seeding)을 하고, 12시간 안정화 후 파쇄상등 sample을 전처리(24h)한 뒤 serum free media로 변화시켰다. alpha MSH로 2일 동안 색소침착을 유도한 후 1N NaOH로 용해시켰다.After inoculating the melan-a cells and stabilizing the samples for 12 hours after stabilization, the samples were changed to serum free media. Pigmentation was induced for 2 days with alpha MSH and then dissolved with 1N NaOH.
멜라닌을 분비하는 cell line에서 유산균 활성성분에 의한 멜라닌 생성의 억제를 확인하였고, 파쇄 정도에 따라 억제가 증가됨을 확인하였다. 이는 미백효과의 증가라고 할 수 있다. 파쇄에 의해 얻을 수 있는 활성성분을 이용한 피부세포에서의 활성은 기존의 유산균이 가지는 활성을 미립화를 통하여, 증대시킬 수 있다. 수용성 활성성분의 활용 분야의 다양성을 증명하는 결과라고 할 수 있다(도 7 참조).In the cell line secreting melanin, the inhibition of melanin production by the lactic acid bacteria active ingredient was confirmed, and the inhibition was increased according to the degree of crushing. This can be said to increase the whitening effect. Activity in skin cells using the active ingredient obtained by crushing can increase the activity of the existing lactic acid bacteria through atomization. It can be said that the results prove the diversity of the field of application of water-soluble active ingredient (see Fig. 7).
<실시예 3> 항 노화 효과Example 3 Anti Aging Effect
human normal dermal Fibroblast cell을 2×105cells/ml로 접종을 하고, 12시간 안정화 후 파쇄상등 sample을 전처리(24h)한 뒤 serum free media로 변화시켰다. UV 25mJ/cm2처리 후, 24시간 동안 배양하고, Sandwich ELISA를 통하여 확인하였다.Human normal dermal Fibroblast cells were inoculated at 2 × 10 5 cells / ml, stabilized for 12 hours, and pre-treated with crushed light samples (24h) and then changed to serum free media. After UV 25mJ / cm 2 treatment, it was incubated for 24 hours, and confirmed by Sandwich ELISA.
UV에 의해 증가되는 MMP-1의 발현의 감소를 통하여 광노화 억제 정도 확인하였다. 파쇄에 의해 만들어지는 유산균 수용성 활성성분에 의한 피부노화 억제의 증가는 파쇄 정도에 의해 유도되는 활성물질의 증가 및 유산균이 가지는 피부 노화억제의 활성을 증대시킬 수 있었다.The degree of photoaging inhibition was confirmed through the decrease of the expression of MMP-1 increased by UV. Increasing the skin aging inhibition by the lactic acid bacteria water-soluble active ingredient produced by the crushing was able to increase the active material induced by the degree of crushing and increase the skin aging inhibitory activity of the lactic acid bacteria.
본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시예들은 한정적인 관점이 아니라 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특허청구범위에 나타나 있으며, 그와 동등한 범위 내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Claims (12)

  1. 유산균 파쇄 및 추출을 통한 수용성 활성성분을 포함하는 항 염증, 피부 미백 및 항 노화용 조성물.Anti-inflammatory, skin whitening and anti-aging composition comprising a water-soluble active ingredient through lactic acid bacteria disruption and extraction.
  2. 제 1항에 있어서, 상기 유산균은 락토바실러스, 비피도박테리움, 스트렙토코커스 또는 락토코커스인 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물. According to claim 1, wherein the lactic acid bacteria Lactobacillus, Bifidobacterium, Streptococcus or lactococcus, characterized in that the composition for anti-inflammatory, skin whitening and anti-aging.
  3. 제 2항에 있어서, 상기 락토바실러스는 락토바실러스 플랜타륨(Lactobacillus plantarum), 락토바실러스 람노서스(Lactobacillus rhamnosus), 락토바실러스 사케이(lactobacillus sakei), 락토바실러스 델브르키(Lactobacillus delbrueckii), 락토바실러스 엑시도필러스(Lactobacillus acidophilus), 락토바실러스 가세리 (Lactobacillus gasseri), 락토바실러스 존스니(Lactoacillus johnsonni), 락토바실러스 헬베티쿠스(Lactobacillus helveticus) 또는 락토바실러스 카세이(Lactobacillus casei)인 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물. The method of claim 2, wherein the Lactobacillus Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus rhamnosus (Lactobacillus rhamnosus), Lactobacillus sakei (lactobacillus sakei), Lactobacillus delbreckii (Lactobacillus delbrueckii), Lactobacillus ecdosi Lactobacillus acidophilus, Lactobacillus gasseri, Lactoacillus johnsonni, Lactobacillus helveticus or Lactobacillus casei, Lactobacillus casei Skin whitening and anti aging composition.
  4. 제 2항에 있어서, 상기 비피도박테리움은 비피도박테리움 비피도(Bifidobacterium bifido), 비피도박테리움 롱검(Bifidobacterium longum), 비피도박테리움 인판티스(Bifidobacterium infantis) 또는 비피도박테리움 아니말리스(bifidobacterium animalis)인 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물. The method of claim 2, wherein the Bifidobacterium bifidobacterium bifido, Bifidobacterium longum, Bifidobacterium infantis or Bifidobacterium infantis (bifidobacterium animalis), characterized in that the composition for anti-inflammatory, skin whitening and anti-aging.
  5. 제 2항에 있어서, 상기 스트렙토코커스는 스트렙토코커스 써모필러스(Streptococcus thermophilus)인 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물. The composition for anti-inflammatory, skin lightening and anti aging according to claim 2, wherein the Streptococcus is Streptococcus thermophilus.
  6. 제 2항에 있어서, 상기 락토코커스는 락토코커스 락티스 (Lactococcus lactis)인 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물. The composition for anti-inflammatory, skin whitening and anti-aging according to claim 2, wherein the lactococcus is Lactococcus lactis.
  7. 제 1항에 있어서, 상기 수용성 활성성분은 유산균 파쇄물의 상등액인 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물.The anti-inflammatory, skin whitening and anti-aging composition according to claim 1, wherein the water-soluble active ingredient is a supernatant of lactic acid bacteria lysate.
  8. 제 7항에 있어서, 상기 수용성 활성 성분의 분자량은 391 내지 597 m/z를 포함하는 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물.8. The composition for anti-inflammatory, skin whitening and anti aging according to claim 7, wherein the water-soluble active ingredient has a molecular weight of 391 to 597 m / z.
  9. 제 7항에 있어서, 상기 유산균 파쇄물의 크기는 100 내지 900nm인 것을 특징으로 하는 항 염증, 피부 미백 및 항 노화용 조성물. 8. The composition for anti-inflammatory, skin whitening and anti aging according to claim 7, wherein the lactic acid bacteria lysate has a size of 100 to 900 nm.
  10. 제 1항 내지 제 8항 중 어느 한 항에 있어서, 피부 외용제로 사용하는 것인 항 염증, 피부 미백 및 항 노화용 조성물.The composition for anti-inflammatory, skin whitening and anti-aging according to any one of claims 1 to 8, which is used as an external preparation for skin.
  11. 제 1항 내지 제 8항 중 어느 한 항에 있어서, 화장료로 사용하는 것인 항 염증, 피부 미백 및 항 노화용 조성물.The composition for anti-inflammatory, skin whitening and anti-aging according to any one of claims 1 to 8, which is used as a cosmetic.
  12. 제 1항 내지 제 8항 중 어느 한 항에 있어서, 건강식품으로 사용하는 것인 항 염증, 피부 미백 및 항 노화용 조성물.The composition for anti-inflammatory, skin whitening and anti-aging according to any one of claims 1 to 8, which is used as a health food.
PCT/KR2013/003158 2012-04-13 2013-04-15 Anti-inflammatory, skin whitening and anti-aging composition comprising supernatant of lactic acid bacteria lysate WO2013154407A1 (en)

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