JP2007070305A - Hair growth/restore agent - Google Patents
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- JP2007070305A JP2007070305A JP2005261218A JP2005261218A JP2007070305A JP 2007070305 A JP2007070305 A JP 2007070305A JP 2005261218 A JP2005261218 A JP 2005261218A JP 2005261218 A JP2005261218 A JP 2005261218A JP 2007070305 A JP2007070305 A JP 2007070305A
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Abstract
Description
本発明は優れた養毛・育毛効果を有する養毛・育毛剤に関する。 The present invention relates to a hair nourishing and hair restoring agent having excellent hair nourishing and hair restoring effects.
養毛・育毛を目的とした医薬品または薬用化粧料には、養毛・育毛効果を有する合成物質又は天然の動植物や微生物の抽出エキス等の薬効成分が広く使用されている。例えば、トランス-3,4’-ジメチル-3-ヒドロキシフラバノンに代表されるフラバノノール誘導体（特許文献１）、Eucalyptus属植物の抽出物（特許文献２）、ストレプトコッカス属やラクトバチルス属細菌の培養濾液（特許文献３）に育毛作用があることが報告されている。しかし、さらに効果の高い養毛・育毛活性を有する剤が求められている。
本発明は、症状や体質に関らず、養毛・育毛を促進し、種々の脱毛症に有効な養毛・育毛剤を提供することにある。 An object of the present invention is to provide a hair nourishing and hair restoring agent effective for various alopecias by promoting hair nourishing and hair restoration regardless of symptoms and constitution.
本発明者は、動植物や微生物等の天然物由来の抽出物から養毛・育毛効果の高い抽出物を探索したところ、乳酸菌の培養液の有機溶媒抽出画分に優れた養毛・育毛効果があることを見出した。また、優れた養毛・育毛効果を有する新規菌株を見出した。 The present inventor searched for an extract having a high hair-restoration / hair-growth effect from extracts derived from natural products such as animals and plants and microorganisms. I found out. Moreover, the novel strain which has the outstanding hair-restoration and hair-growth effect was discovered.
すなわち本発明は、乳酸菌培養液の有機溶媒抽出液を有効成分とする養毛・育毛剤を提供するものである。 That is, the present invention provides a hair nourishing and hair restorer comprising an organic solvent extract of lactic acid bacteria culture as an active ingredient.
また、本発明は、乳酸菌培養液の有機溶媒抽出物を含有する養毛・育毛剤であって、乳酸菌培養液の有機溶媒抽出物による養毛・育毛効果がある旨を表示した養毛・育毛剤を提供するものである。 Further, the present invention is a hair nourishing / hair restoring agent containing an organic solvent extract of a lactic acid bacteria culture solution, and displays that there is a hair nourishing / hair restoring effect by the organic solvent extract of the lactic acid bacteria culture solution. An agent is provided.
また、本発明は乳酸菌の培養液を有機溶媒抽出することを特徴とする養毛・育毛剤の製造方法を提供するものである。 The present invention also provides a method for producing a hair nourishing / hair-growing agent characterized by extracting a culture solution of lactic acid bacteria with an organic solvent.
さらに、本発明は、独立行政法人産業技術総合研究所 特許微生物寄託センターにＦＥＲＭ Ｐ−２０５８６として寄託された微生物（ロイコノストック・メセンテロイデスＫＳＭ−４／１０８）を提供するものである。 Furthermore, the present invention provides a microorganism (Leuconostoc mesenteroides KSM-4 / 108) deposited as FERM P-20586 at the Patent Microorganism Depositary, National Institute of Advanced Industrial Science and Technology.
本発明の養毛・育毛剤を頭皮等に適用することにより、適用部位において優れた養毛・育毛効果が発揮される。 By applying the hair nourishing and hair restoring agent of the present invention to the scalp and the like, an excellent hair nourishing and hair restoring effect is exhibited at the application site.
本発明に用いる乳酸菌としては、その有機溶媒抽出物が養毛・育毛活性を有するものであれば限定はされないが、ラクトバチルス（Lactobacillus）属、ペディオコッカス(Pediococcus)属又はロイコノストック(Leuconostoc)属由来の乳酸菌が好ましい。特に、ラクトバチルス・ラクティス(Lactobacillus lactis)JCM1107、ペディオコッカス・アシディラクティシィ (Pediococcus acidilactici)JCM9797又は、ロイコノストック・メセンテロイデス(Leuconostoc mesenteroides)KSM-4/108が好ましい。 The lactic acid bacterium used in the present invention is not limited as long as the organic solvent extract has hair-growing / hair-growing activity, but is not limited to Lactobacillus , Pediococcus or Leuconostoc. ) -Derived lactic acid bacteria are preferred. In particular, Lactobacillus lactis JCM1107, Pediococcus acidilactici JCM9797 or Leuconostoc mesenteroides KSM-4 / 108 is preferable.
上記乳酸菌のうち、ロイコノストック・メセンテロイデス(Leuconostoc mesenteroides)KSM-4/108は、発明者らにより市販の白菜漬けより分離された菌株である。以下に当該菌株の菌学的性質並びに生理学的性質について示す。
[KSM-4/108の性質]
Ａ．形態学的性質
（１）形態 MRS寒天平板培地を用いて30℃、48時間嫌気培養後の結果を示す。形状：球菌、大きさ：0.5-1×1-2μm、連鎖したもの多数
（２）グラム染色性：陽性
（３）コロニー形態 形状：円形、周縁：全縁、大きさ：直径0.2-0.8mm、色調：白色、表面：円滑
（４）芽胞形成：陰性
（５）運動性：なし
Ｂ．生理学的性質
（１）ガス産生：あり
（２）カタラーゼ活性：陰性
（３）脱脂乳凝固性：凝固
（４）ゼラチン液化性：なし
（５）硝酸塩還元性：なし
（６）インドール産性：なし
（７）硫化水素産性：なし
（８）糖の発酵性：市販の細菌同定用キットAPI 50 CHL（bioMerieux社）にて糖の発酵性を検討した結果を以下に記載する。（＋は発酵性有りを示し、−は発酵性なしを示す。）
グリセロール −
エリスリトール −
D-アラビノース −
L-アラビノース ＋
リボース ＋
D-キシロース ＋
L-キシロース −
アドニトール −
β-メチル-D-キシロシド −
ガラクトース ＋
D-グルコース ＋
L-フルクトース ＋
D-マンノース −
L-ソルボース −
ラムノース −
ダルシトール −
イノシトール −
マンニトール −
ソルビトール −
α-メチル-D-マンノシド −
α-メチル-D-グルコシド ＋
N-アセチル-グルコサミン ＋
アミグダリン−
アルブチン−
エスクリン ＋
サリシン−
セロビオース−
マルトース ＋
ラクトース ＋
メリビオース＋
サッカロース ＋
トレハロース ＋
イヌリン −
メレジトース −
D-ラフィノース ＋
スターチ −
グリコーゲン −
キシリトール −
β-ゲンチオビオース −
D-ツラノース ＋
D-リキソース −
D-タガトース −
D-フコース −
L-フコース −
D-アラビトール −
L-アラビトール −
グルコネート −
２-ケト-グルコネート −
５-ケト-グルコネート −
Among the above lactic acid bacteria, Leuconostoc mesenteroides KSM-4 / 108 is a strain isolated from commercially available Chinese cabbage pickled by the inventors. The bacteriological and physiological properties of the strain are shown below.
[Properties of KSM-4 / 108]
A. Morphological properties (1) Morphology The results after anaerobic culture at 30 ° C. for 48 hours using MRS agar plates are shown. Shape: cocci, size: 0.5-1 × 1-2 μm, many linked (2) Gram staining: positive (3) colony shape Shape: circular, peripheral edge: full edge, size: 0.2-0.8 mm in diameter, Color: White, Surface: Smooth (4) Spore formation: Negative (5) Motility: None Physiological properties (1) Gas production: Yes (2) Catalase activity: Negative (3) Nonfat milk coagulation: Coagulation (4) Gelatin liquefaction: No (5) Nitrate reduction: No (6) Indole productivity: No (7) Hydrogen sulfide productivity: None (8) Sugar fermentability: The results of examination of sugar fermentability with a commercially available bacterial identification kit API 50 CHL (bioMerieux) are described below. (+ Indicates that there is fermentability, − indicates that there is no fermentability.)
Glycerol −
Erythritol −
D-arabinose −
L-arabinose +
Ribose +
D-xylose +
L-xylose −
Adonitol −
β-methyl-D-xyloside −
Galactose +
D-glucose +
L-fructose +
D-Mannose −
L-sorbose −
Rhamnose −
Dulcitol −
Inositol −
Mannitol −
Sorbitol −
α-Methyl-D-mannoside −
α-methyl-D-glucoside +
N-acetyl-glucosamine +
Amygdalin
Arbutin
Esclin +
Salicin-
Cellobiose
Maltose +
Lactose +
Melibiose +
Sucrose +
Trehalose +
Inulin −
Merezitose −
D-Raffinose +
Starch −
Glycogen −
Xylitol −
β-gentiobiose −
D-Turanorth +
D-lyxose −
D-Tagatose −
D-Fucose −
L-Fucose −
D-arabitol −
L-arabitol −
Gluconate −
2-keto-gluconate −
5-keto-gluconate −
以上の菌学的性状は、典型的なLeuconostoc mesenteroidesの性状を示し、糖の発酵性は、Leuconostoc mesenteroides subsp. dextranicum 2と９３．２％の相同率を示した。
次いで、4/108株のゲノムＤＮＡの内、16S rDNA-Full塩基配列を解析した結果、相同率１００％で既存株と一致したので4/108株は、Leuconostoc mesenteroidesに帰属する菌株と推定した。そこで、本発明者は、本発明の菌株をロイコノストック・メセンテロイデス（Leuconostoc mesenteroides KSM-4/108）と命名し、同微生物を2005年7月5日付けで独立行政法人産業技術総合研究所特許微生物寄託センターに寄託番号FERM P-20586として寄託した。
The above bacteriological properties showed typical Leuconostoc mesenteroides properties, and the sugar fermentability showed 93.2% homology with Leuconostoc mesenteroides subsp. Dextranicum 2.
Next, as a result of analyzing the 16S rDNA-Full nucleotide sequence in the genomic DNA of 4/108 strain, 4/108 strain was estimated to be a strain belonging to Leuconostoc mesenteroides because it matched the existing strain with a homology rate of 100%. Therefore, the present inventor named the strain of the present invention Leuconostoc mesenteroides KSM-4 / 108, and the microorganism was named as an independent administrative institute of National Institute of Advanced Industrial Science and Technology on July 5, 2005. Deposited at the Microorganism Depositary as deposit number FERM P-20586.
本発明の育毛・養毛剤やその製造方法に使用する乳酸菌の培養液を取得するには、通常使用される乳酸菌用の培地に上記の乳酸菌を摂取し、常法に従って培養すればよい。ここで培地としては、液体培地が好ましく、例えば、ＭＲＳブロス（OXOID）や一般乳酸菌接種用培地（日水製薬）が挙げられる。培養は、２０〜４０℃で、２４〜７２時間行うのが好ましく、培養後、遠心分離や濾過により菌体を除くことで、有機溶媒抽出用の培養液とすることができる。 In order to obtain the culture solution of lactic acid bacteria used in the hair growth / hair growth agent of the present invention and the method for producing the same, the above-mentioned lactic acid bacteria may be ingested into a commonly used medium for lactic acid bacteria and cultured according to a conventional method. Here, the medium is preferably a liquid medium, and examples thereof include MRS broth (OXOID) and a medium for inoculating general lactic acid bacteria (Nissui Pharmaceutical). Cultivation is preferably performed at 20 to 40 ° C. for 24 to 72 hours. After culturing, the cells can be removed by centrifugation or filtration to obtain a culture solution for organic solvent extraction.
上記培養液の抽出に用いる有機溶媒としては、ペンタン、ヘキサン、ヘプタン、オクタン等の飽和炭化水素、あるいはヘキセン、ヘプテン等の不飽和炭化水素や、メタノール、エタノール、１-プロパノール等のアルコール類、アセトン、酢酸エチルが挙げられるが、非極性溶媒が好ましく、ヘキサンが特に好ましい。 Examples of the organic solvent used for the extraction of the culture broth include saturated hydrocarbons such as pentane, hexane, heptane, and octane, unsaturated hydrocarbons such as hexene and heptene, alcohols such as methanol, ethanol, and 1-propanol, acetone. Ethyl acetate, nonpolar solvents are preferred, and hexane is particularly preferred.
抽出条件は、使用する溶媒によっても異なるが、例えばヘキサンにより抽出する場合、上記菌体を除いた培養液の0.5〜10倍量のヘキサンを用い、5〜40℃、好ましくは10〜30℃の温度で、5分〜3時間、より好ましくは10分〜1時間抽出することが好ましい。 Extraction conditions vary depending on the solvent to be used. For example, when extracting with hexane, 0.5 to 10 times the amount of hexane except for the above-mentioned bacterial cells is used, and 5 to 40 ° C, preferably 10 to 30 ° C It is preferable to extract at a temperature of 5 minutes to 3 hours, more preferably 10 minutes to 1 hour.
上記の抽出物は、そのまま用いることもできるが、当該抽出物を希釈、濃縮若しくは凍結乾燥した後、必要に応じて粉末又はペースト状に調整したり、エタノール等の別の溶媒に溶解したりして使用することもできる。
また、液々分配等の技術により、上記抽出物から不活性な夾雑物を除去して用いることもでき、本発明においてはこのようなものを用いることが好ましい。これらは必要により、公知の方法で脱臭、脱色等の処理を施してからでも良い。
The above extract can be used as it is, but after diluting, concentrating or lyophilizing the extract, it can be adjusted to powder or paste as necessary, or dissolved in another solvent such as ethanol. Can also be used.
In addition, inert impurities can be removed from the extract by a technique such as liquid-liquid distribution, and it is preferable to use such a material in the present invention. These may be subjected to treatment such as deodorization and decolorization by a known method, if necessary.
本発明の乳酸菌培養液の有機溶媒抽出液は、後記実施例に示すように、優れた養毛・育毛活性を有することから、養毛・育毛効果を奏する医薬部外品、医薬品、毛髪用化粧料等として使用可能な養毛・育毛剤とすることができる。本発明の養毛・育毛剤の投与形態としては、経口剤、非経口剤の何れでもよいが、外用剤が好ましい。なお、本発明において養毛・育毛は、養毛及び／又は育毛を意味し、これには発毛および脱毛予防の概念が包含されるものとする。 The organic solvent extract of the lactic acid bacterium culture solution of the present invention has excellent hair-restoring / hair-growth activity as shown in the examples below, so that it is a quasi-drug, pharmaceutical, and hair makeup that exhibits hair-growth / hair-growth effects. It can be used as a hair nourishing and hair-restoring agent that can be used as a material. The dosage form of the hair nourishing / hair-growing agent of the present invention may be either an oral preparation or a parenteral preparation, but an external preparation is preferred. In the present invention, hair growth / hair growth means hair growth and / or hair growth, which includes the concept of hair growth and hair loss prevention.
本発明の養毛・育毛剤は、その剤形は毛髪及び頭皮に、効果的に塗布される剤形が好ましく、通常は、液剤を主体とし、ローションやヘアトニックが代表的なものであり、噴射剤と共にエアゾール剤として使用できる。また、クリームやジェル、シャプー、リンス、ヘアフォーム剤、ヘアスプレー剤等の剤形で使用することもできる。使用の簡便さからエアゾール剤とすることが好ましい。 The hair nourishing / hair-growth agent of the present invention is preferably a dosage form that is effectively applied to the hair and scalp, and is typically a liquid agent, typically lotions and hair tonics, It can be used as an aerosol together with a propellant. Moreover, it can also be used in dosage forms such as cream, gel, shampoo, rinse, hair foam, hair spray. From the viewpoint of ease of use, an aerosol agent is preferred.
エアゾール剤に使用する噴射剤としては、炭酸ガス、ＬＰＧ、ＤＭＥ、窒素ガス、イソペンタン等を１種または２種以上組み合わせて用いることができ、特に、使用感の面から炭酸ガスを含有させることが好ましい。また、炭酸ガスは、頭皮中の毛細血管の血行を促進する効果を有し、本発明の養毛・育毛促進効果をさらに増加させることができる。 As the propellant used for the aerosol agent, carbon dioxide, LPG, DME, nitrogen gas, isopentane or the like can be used alone or in combination of two or more, and in particular, carbon dioxide may be contained from the viewpoint of feeling of use. preferable. Carbon dioxide has an effect of promoting blood circulation in the capillaries in the scalp, and can further increase the effect of promoting hair restoration / hair growth of the present invention.
炭酸ガスは、これが溶解している溶液のｐＨが酸性の場合にはCO2分子として存在し、血管拡張作用を示す。従って、炭酸ガスを含有した本発明の養毛・育毛剤の液性はｐＨ７以下、特にｐＨ４．５〜６．５に調整するのが好ましい。尚、炭酸ガスは養毛・育毛剤中に溶け込むと酸性度が高くなるが、必要に応じて最終ｐＨが上記範囲内になる様に調節すれば良い。このｐＨ調整剤としては、例えば、クエン酸、酒石酸、乳酸等の有機酸またはこれらの塩、或いはリン酸またはその塩が好適に使用される。 Carbon dioxide gas exists as CO 2 molecules when the pH of the solution in which it is dissolved is acidic, and exhibits a vasodilating action. Accordingly, the liquidity of the hair nourishing and hair restoring agent of the present invention containing carbon dioxide is preferably adjusted to pH 7 or less, particularly pH 4.5 to 6.5. Carbonic acid gas increases in acidity when dissolved in a hair nourishing / hair-growing agent, but may be adjusted so that the final pH is within the above range, if necessary. As this pH adjuster, for example, organic acids such as citric acid, tartaric acid and lactic acid or salts thereof, or phosphoric acid or salts thereof are preferably used.
上記エアゾール剤の製造方法は、例えば、炭酸ガスを除く他の成分を耐圧容器に入れ、これに高圧炭酸ガスを封入する方法、耐圧容器に炭酸水素ナトリウム等の炭酸塩を含ませた炭酸ガスを発生する成分を入れ、これに適当なｐＨ調整剤を加えて炭酸ガスを発生させ、直ちに密封する方法、或いはドライアイスペレットを容器内に入れて密封する方法等が採用されるが、特に高圧炭酸ガスを封入する方法が好ましい。 The method for producing the aerosol agent includes, for example, a method in which other components excluding carbon dioxide gas are put in a pressure vessel, and high pressure carbon dioxide gas is sealed in the pressure vessel. A method of adding a component to be generated and adding a suitable pH adjuster to generate carbon dioxide and immediately sealing it, or a method of sealing by putting dry ice pellets in a container is adopted. A method of enclosing gas is preferred.
斯くして得られる本発明のエアゾール剤は、炭酸ガスの一部が養毛・育毛剤中に溶解して存在し、また、一部は容器中に気体として存在するが、特に、炭酸ガスが養毛・育毛剤中に溶けて配合されていることが重要であり、この含有量は炭酸ガス質量濃度が６０ppm以上であることが好ましい。炭酸ガスの含有量の調節は、炭酸ガスの注入（圧入）によって行うことができ、一般に容器内圧力が３５℃の温度で１．２〜１０kg/cm2（ゲージ圧）になるようにするのが好ましく、特に４．５〜１０kg/cm2（ゲージ圧）になるようにするのが好ましい。 In the aerosol agent of the present invention thus obtained, a part of carbon dioxide gas is dissolved in the hair nourishing / hair restoring agent, and a part thereof is present as a gas in the container. It is important that it is dissolved and blended in the hair nourishing agent, and this content is preferably such that the carbon dioxide mass concentration is 60 ppm or more. The content of carbon dioxide can be adjusted by injecting (press-fitting) carbon dioxide. Generally, the pressure in the container is 1.2 to 10 kg / cm 2 (gauge pressure) at a temperature of 35 ° C. In particular, it is preferable that the pressure be 4.5 to 10 kg / cm 2 (gauge pressure).
本発明の養毛・育毛剤は、本発明の効果を損なわない範囲において、乳酸菌培養液の有機溶媒抽出物のほかに、通常用いられる養毛薬効剤、例えば、抗炎症剤、細胞不活化剤、皮脂分泌抑制剤、抹消血管拡張剤、アミノ酸類、ビタミン類等を必要に応じて適宜配合し、養毛・育毛効果の向上を図ることができる。また、その他化粧品、医薬部外品、医薬品等の成分として一般に使用されている成分を必要に応じて適宜配合することが出来る。この様な任意成分としては、例えば、精製水、エタノール、油性物質、保湿剤、増粘剤、防腐剤、乳化剤、薬効成分、粉体、紫外線吸収剤、色素、香料、乳化安定化剤等を任意に組み合わせて配合することができる。 As long as the effect of the present invention is not impaired, the hair nourishing / hair-growing agent of the present invention is used in addition to the organic solvent extract of the lactic acid bacteria culture solution, and usually used hair nourishing agents such as anti-inflammatory agents and cell inactivating agents. In addition, a sebum secretion inhibitor, a peripheral vasodilator, amino acids, vitamins, and the like can be appropriately blended as necessary to improve the hair restoration / hair growth effect. In addition, other components generally used as components of cosmetics, quasi drugs, pharmaceuticals, and the like can be appropriately blended as necessary. Examples of such optional components include purified water, ethanol, oily substances, moisturizers, thickeners, preservatives, emulsifiers, medicinal ingredients, powders, ultraviolet absorbers, dyes, fragrances, and emulsion stabilizers. Any combination can be added.
本発明の養毛・育毛剤における乳酸菌培養液の有機溶媒抽出物の含有量は、添加形態および投与形態によっても異なるが、広い範囲から選択できる。例えば、溶媒抽出乾燥物換算で、組成物中に0.0001質量％〜0.1質量％配合することが好ましく、特に0.001質量％〜0.01質量％とすることが好ましい。 The content of the organic solvent extract of the lactic acid bacteria culture solution in the hair nourishing / hair-growing agent of the present invention can be selected from a wide range, although it varies depending on the addition form and administration form. For example, it is preferable to mix 0.0001 mass%-0.1 mass% in a composition in conversion of a solvent extraction dry matter, and it is preferable to set it as 0.001 mass%-0.01 mass% especially.
以下、実施例を挙げて本発明を詳細に説明する。
実施例１ 漬物からLeuconostoc mesenteroides KSM-4/108株の分離
栃木県内で市販されている白菜漬けを入手し、その滲出液1mLを滅菌生理食塩水で10-1〜10-7まで段階的に希釈した後、各希釈液0.1mLをBCP寒天培地（Difco）に接種し、酸素吸収・炭酸ガス発生剤（Anaero Pack、三菱ガス化学(株)社製）で嫌気状態に保った酸素不透過性のビニール袋内にて30℃で72時間嫌気培養した。生じた黄色のコロニーを滅菌生理食塩水に懸濁し、この懸濁液をBCP寒天培地で嫌気培養する事を繰り返す事によって分離菌株を純化した。純化した各々の分離菌株に関し、グラム染色性陽性、胞子形成なし、運動性なし、カタラーゼ陰性等の確認を行い、乳酸菌として保存した。この様にして得られた乳酸菌の中から、実施例２の方法によって培養し、得られた培養液の有機溶媒抽出物の養毛・育毛活性の高い株をLeuconostoc mesenteroides KSM-4/108株（FPRM P-20586）として選抜した。
Hereinafter, the present invention will be described in detail with reference to examples.
Example 1 Separation of Leuconostoc mesenteroides KSM-4 / 108 strain from pickles Obtained Chinese cabbage pickles marketed in Tochigi Prefecture and diluted 1 mL of the exudate in a stepwise manner from 10 -1 to 10 -7 with sterile physiological saline. After that, 0.1 mL of each diluted solution was inoculated into BCP agar medium (Difco) and kept in anaerobic condition with oxygen absorption / carbon dioxide generator (Anaero Pack, manufactured by Mitsubishi Gas Chemical Co., Ltd.). Anaerobic culture was carried out at 30 ° C. for 72 hours in a plastic bag. The resulting yellow colony was suspended in sterilized physiological saline, and this suspension was anaerobically cultured on a BCP agar medium to purify the isolated strain. Each purified isolate was checked for positive Gram staining, no sporulation, no motility, negative catalase, etc., and stored as lactic acid bacteria. The lactic acid bacteria thus obtained were cultured by the method of Example 2, and an organic solvent extract of the obtained culture broth was used as a strain of Leuconostoc mesenteroides KSM-4 / 108 ( FPRM P-20586).
実施例２ 各種乳酸菌培養液の有機溶媒抽出物の調製
市販の乳酸菌からラクトバチルス・ラクティス(Lactobacillus lactis)JCM1107、ペディオコッカス・アシディラクティシィ (Pediococcus acidilactici)JCM9797の2株、及び、ロイコノストック・メセンテロイデス(Leuconostoc mesenteroides)KSM-4-108の3菌株を供試菌株として用いた。ＭＲＳ寒天培地に生育させた新鮮な各菌体を１００ｍLのＭＲＳブロス培地に接種して、３０℃で３日間静置培養した。培養液を遠心分離する事によって得られた培養上清に等量のヘキサンを添加して攪拌し、ヘキサン相を濃縮乾固したものをヘキサン抽出物とし、５ｍLのエタノールに溶解したものを乳酸菌培養エキスとした。MRSブロス培地１００ｍL中のヘキサン抽出物の乾燥重量を表1に示す。（表１）。
Example 2 Preparation of Organic Solvent Extracts of Various Lactic Acid Bacteria Culture Solutions From commercially available lactic acid bacteria, two strains of Lactobacillus lactis JCM1107 and Pediococcus acidilactici JCM9797, and Leuconostoc -Three strains of Leuconostoc mesenteroides KSM-4-108 were used as test strains. Each fresh cell grown on the MRS agar medium was inoculated into 100 mL of MRS broth medium and statically cultured at 30 ° C. for 3 days. An equal amount of hexane is added to the culture supernatant obtained by centrifuging the culture solution and stirred, and the hexane phase is concentrated to dryness to obtain a hexane extract, which is dissolved in 5 mL of ethanol and cultured in lactic acid bacteria. It was an extract. Table 1 shows the dry weight of the hexane extract in 100 mL of MRS broth medium. (Table 1).
実施例３ 養毛・育毛活性の評価
生後５日のＳＤ系ラットの髭毛包器官を外科的に摘出した。器官培養用シャーレ（Ｆａｌｃｏｎ ３０３７）の外周部分に湿度維持のためのリン酸緩衝液を入れ、内皿上に置いた三角グリット上に得られた毛包（５本／dish）を並べた。無血清培地（ＲＰＭＩ １６４０）中に、実施例２で調製した抽出物を最終濃度０．５％となるように加え、ＣＯ2の気相下にて、３７℃で培養し、経日的毛伸長を観察した。その結果を表２に示す。表２中の毛伸長度は、培地コントロール群を１００とした場合の培養４日目の相対値である。表２から明らかなように、乳酸菌培養上清のヘキサン抽出画分に顕著な毛幹伸長促進効果が認められた。
Example 3 Evaluation of hair restoration / hair growth activity The eyelash follicle organs of 5 days old SD rats were surgically removed. A phosphate buffer for maintaining humidity was placed in the outer periphery of an organ culture petri dish (Falcon 3037), and the hair follicles (5 / dish) obtained were placed on a triangular grit placed on an inner dish. The extract prepared in Example 2 was added to a serum-free medium (RPMI 1640) to a final concentration of 0.5%, and cultured at 37 ° C. in a gas phase of CO 2. Elongation was observed. The results are shown in Table 2. The degree of hair elongation in Table 2 is the relative value on the fourth day of culture when the medium control group is 100. As is clear from Table 2, a remarkable hair shaft elongation promoting effect was observed in the hexane extract fraction of the lactic acid bacteria culture supernatant.
配合例１ 養毛・育毛剤１
Ａ）
Formulation Example 1 Hair nourishing and hair restorer 1
A)
Ａに属する成分を溶解し、別にＢに属する成分を溶解し、ＡにＢを添加して均一に攪拌混合し、養毛・育毛剤を得た。 A component belonging to A was dissolved, another component belonging to B was dissolved, B was added to A, and the mixture was stirred and mixed uniformly to obtain a hair nourishing / hair growth agent.
配合例２ 養毛・育毛剤２
Ａ）
Formulation Example 2 Hair nourishing and hair restorer 2
A)
Ａに属する成分を均一に混合し容器に入れ、常法によりＢを容器に充填し、炭酸ガスを含有する養毛・育毛剤を得た。 Ingredients belonging to A were uniformly mixed and placed in a container, and B was filled into the container by a conventional method to obtain a hair nourishing and hair restorer containing carbon dioxide gas.
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| JP2015096475A (en) * | 2013-11-15 | 2015-05-21 | 株式会社東洋新薬 | Composition for collagen production promotion |
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| JP2015187171A (en) * | 2015-07-16 | 2015-10-29 | 国立研究開発法人農業・食品産業技術総合研究機構 | External preparation for skin for hair restoration/growth using lactic acid bacteria |
| JP2018035136A (en) * | 2016-08-30 | 2018-03-08 | コエンバイオ カンパニー リミテッド | Composition having lipolysis ability for alopecia prevention or for hair growth promotion comprising the strain which possesses |
| CN110225969A (en) * | 2016-08-30 | 2019-09-10 | (株)库恩生物 | Have the effect of preventing hair loss, promote hair growth or improving the Zha Feier leukonid of sexual function and the composition including it |
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| CN110325197A (en) * | 2016-08-30 | 2019-10-11 | (株)库恩生物 | For prevent hair loss or it is trichogenous include show lipolysis effect bacterial strain composition |
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| EP3816304A3 (en) * | 2016-08-30 | 2021-07-21 | Coenbio Co., Ltd. | Composition, for preventing hair loss or promoting hair growth, comprising strains showing lipolysis effect |
| CN110225969B (en) * | 2016-08-30 | 2023-07-11 | (株)库恩生物 | Leuconostoc zafii and culture solution, concentrate, dried product, food composition and pharmaceutical composition comprising same |
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